目的:探讨血清、尿金属蛋白酶组织抑制因子-1(TIMP-1)浓度对肾脏纤维化的诊断价值.方法: 以酶联免疫吸附检测(ELISA)方法检测健康者及肾脏病患者血清、尿TIMP-1浓度.结果: ①以血清TIMP-1浓度显著增高来判断肾组织有无纤维化敏感性为89%,特异性为94%,与肾活检符合率为85%.②尿TIPM-1浓度与尿蛋白呈正相关(r=0.53,P＜0.01).结论: 血清TIMP-1浓度可作为判断慢性肾脏病人肾纤维化的无损伤性检查指标.
作者：汤王旬；袁发焕 刊期： 2001年第08期
AIM: Shengdeye is a kind of medicinal wine. It is fermented with 30 kinds of medicinal herbs such as glossy ganoderma, pilose antler, panax, medlar and sorghum. This study aimed to observe the effects of Shengdeye on anti-fatigue and memory in mice.METHODS: The mice were fed with Shengdeye 5 ml*kg-1 and 2.5 ml*kg-1 2 times per day. After 7 days, the mice were examined on anoxia-resistant and swimming test for anti-fatigue and step down test for memory.RESULTS: The anoxia-resistant of shengdege group was apparently longer than that of the control group ［(29.60±1.36) min vs (24.40±3.13) min, P＜0.01］, and the swimming time was sharply increased ［(142.6±53.8) min for 2.5 ml*kg-1 and (162.9±43.5) min for 5 ml*kg-1 vs (94.9±39.1) min, P＜0.05］. The error times of shengdege group was lower than that of the control ［(1.5±1.4) times and (2.3±1.3) times vs (3.7±1.1) times， P＜0.01］. The delitescence of two group were sharply prolonged ［(136.8±50.1) s, and (128.0±41.5) s vs (50.2±42.9) s， P＜0.01］.CONCLUSION: The results indicated that Shengdeye had an anti-fatigne effect and increased the faculty of memory. The experiment of acute toxicity showed that the medicinal herbs in Shyengdeye were safe.
作者： 刊期： 2001年第08期
AIM: PSP94 has been shown promise as a potential prostate cancer marker and it was reported that PSP94 can inhibit the growth of prostate cancer cell in vitro and in vivo. This study aimed to construct recombinant human PSP94 expression vector. METHODS:The PSP94 cDNA was obtained from normal prostate tissue, and recombinant plasmid pUC19-PSP94 was constructed. The target gene was identified and sequenced. Then the PSP94 gene was inserted to the secretory expression vector. RESULTS:The gene sequence of PSP94 was identified. The recombinant vector was constructed. The secreted PSP94 was isolated and identified by Western blot. CONCLUSION:The recombinated PSP94 could expressed PSP94 successfully.
作者： 刊期： 2001年第08期
This study investigated the protection against the ND in chickens by a recombinant DNA vaccine. A plasmid vector encoding NDV F protein, which is reqired for virus cell fusion and is important for vaccine induced immunity, was used as a model to study how DNA vaccines may be modulated by the simulaneous expression of chicken IL-2. The NDV D26 strain F gene with CMV promotor and BGH polyA signal sequence was amplified by PCR from eukaryotic plasmid pcDNA-F, which contains the full-length NDV F gene, and clond into reconstructed eukaryotic plasmid pcDNA-IL2, which contains chicken IL-2 gene. Restriction endonuclease cleavage and PCR amplification showed that a bicistronic plasmid encoding NDV F gene and chicken IL-2 separately was successfully constructed. Two-week-old SPF chickens were intramuscularly innoculated the recombinant plasmid. Antibody and lymphocyte proliferative assay showed that the humoral and cellular immunity of chickens vaccinated the recombinant plasmid greatly increased compared with those innoculated only plasmid expressing NDV F protein. Challenged with the lethal dose of NDV F48E9 strain, 72% chickens vaccinated recombinant plasmid were survived, and 30% chickens vaccinated plasmid expressing F protein were survived. These results proved the adjuvant effect of chicken IL-2, and further showed that the efficacy of a DNA vaccine can be greatly improved by simultaneous expression of IL-2.
作者： 刊期： 2001年第08期
It is very difficult for doctor to treat patient with persistent hypotension in the late stage of shock. The aim of present study was to elucidate the reason for lower vasoreactivity in severe shock. Irreversible hemorrhagic shock of rat was reproduced and the vasoreactivity of arteriole in spinotrapezius muscle to norepinephrine (NE) was measured. The resting membrane potential of isolated arterial strips was detected with a microelectrode. The effect of NO on the membrane potential and intracellular ［Ca2＋］i level in isolated arteriolar smooth muscle cells (ASMCs) was determined with fluorescent probes under confocal microscope. KATP channel of ASMCs was measured with patch clamp method. It was shown that membrane hyperpolarization appeared in arteroles 2 h post hemorrhage, while the resting potential was increased from (-36.9±6.3) mV of control value to (-51.0±9.1) mV with the NE threshold increased to 15 times more than preshock value. The hyperpolarization of ASMCs was closely related to vascular hyporeactivity (correlation coefficient 0.96, P＜0.01). The hyperpolarization was enhanced by lack of ATP, increase in H＋，and OONO- in ASMCs. Single KATP channel conducatance, mean open time and open probability was increased in ASMCs, and the increased ［Ca2＋］i level of ASMCs stimulated by NE was reduced to 50% of normal value. The vasoreactivity, blood pressure and survival rate could be improved by the treatment of glybenclamide and NaHCO3. The study indicates that hyperpolarization of ASMCs is a major reason for lower vasoreactivity in severe shock, since it inhibits the voltage dependent Ca2＋ channel (POC) with the reduction of NE stimulated ［Ca2＋］i increase. The decrease in ATP and increase in H+, and OONO- in ASMCs involves in the activation of KATP channel, leading to the ASMCs hyperpolarization.
作者： 刊期： 2001年第08期
Xinmailong injection solution was invented at 1988 by prof. Li Shunan in Dali medical college. It was made from the material which has high biological activeness to the cardiac and vascular system. During the experimental shock caused by excessive loss of blood in monkey and dog, it was found by ECG that the T-wave of anterion lead on left chest elevated and became high and sharp after acute blood loss. Arterial blood pressure dropped to 8-5.3 kPa for dog and 8-5.3-2.7 kPa for monkey, changes of T-wave all recovered to near normal level after xinmailong solution was injected intravenously (0.05-0.2 mL/kg). These Results implied that xinmailong might improve the ischemia of myocardium induced by hemorrhagic shock.
作者： 刊期： 2001年第08期
AIM：Damaging of Endothelial cells and adhesion of leukocytes and platelets were studied in rat mesentery microvessels after ischemia-reperfusion.METHODS：The model was made by losing blood and reperfusion from carotid artery in rat. Changes in mesentery microvessels were observed by high magnify microscope.RESULTS:Leukocyte and platelet adhesions were found in venules and co-capillaries 1-3 hours after ischemia-reperfusion. Endothelial cells were edema and vascular walls were thickening. Vacuoles formed in intracytoplasm of some vascular endothelium and some bigger endothelial vacuoles prominenced toward the luminal surface. Vacuoles had the shape of circle and the diameter was 10-30 μm. More vacuoles were found in arterioles, even there were several vacuoles in a arteriole. The biggest vacuole almost occupied 2/3 of vascular lumen.CONCLUSION：Edema and vacuole formation in vascular endothelium showed endothelial cells were damaged seriously after ischemia-reperfusion.
作者： 刊期： 2001年第08期
In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P＜0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P＜0.01 or P＜0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P＜0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.
作者： 刊期： 2001年第08期
To examine anti-obesity and-diabetic effects of neuronal histamine especially in leptin resistant states, we investigated the effects of chronic central treatment with histamine on lipid, glucose and energy metabolism of db/db obese mice, which are genetically leptin receptor mutated mice. Chronic centrally treatment with histamine (0.05 μmol*g-1 body weight*d-1 for 7 days) decreased body weight, food intake in db/db obese mice, and decreased body fat weight, serum concentration of glucose compared with pair-fed db/db obese mice. Neuronal histamine also suppressed ob mRNA in the white adipose tissue (WAT), serum leptin and increased UCPs mRNA expression in brown adipose tissue (BAT) and in WAT compared with pair-fed controls. These above effects of the histamine were attenuated in these mice with combination of targeted disruption of the histamine H1 receptor gene. In conclusion, neuronal histamine can regulate body fat deposition, serum glucose, leptin, BAT and WAT UCPs expression even in leptin insensitive db/db obese mice.
作者： 刊期： 2001年第08期
This study was designed to invesigate vasodilatory action of exogenous peroxynitrite (ONOO-), and effect of endothelial cells on ONOO- -induced relaxation in isolated rabbit pulmonary arterial rings (PARs). Results were as follows: (1) In precontracted PARs, ONOO- could give rise to vasodilation in a dose-dependent manner. Relaxations of PARs to ONOO- at doses of 10-5 mol/L, 5×10-5 mmol/L and 10-4 mol/L were 11.09%±1.84%, 31.10%±3.53% and 64.35%±3.83%, respectively, all of which were significantly higher than those of decomposed ONOO- with 5.88%±1.27%、16.15%±1.82% and 34.44%±3.26% at same concentrations, respectively. (2) Compared with SNP and ACh, ONOO- had weak relaxant action. (3) ONOO- induced more significantly enhanced relaxation in denuded endothelial PARs than in intact endothelial PARs. (4) In this experimental condition, the relaxation of PARs to 10-6 mol/L ACh remained unchanged before and after observation of relaxation to ONOO-. (5) The relaxations of PARs to 5×10-5 mol/L ONOO- in repetitively administered manner appeared progressively decreased. These results suggested that ONOO- might be implicated in pulmonary hypertension in the early stage of endotoxic shock.
作者： 刊期： 2001年第08期
AIM: To investigate the relationship between cytokine level and nitric oxide (NO) content in aqueous humor after intraocular lens implantation. METHODS: All New Zealand rabbits were divided randomly into three groups: (1) control group, (2) extracapsular cataract extraction group (ECCE), (3) extracapsular cataract extraction and posterior chamber intraocular lens implantation group (ECCE+IOL). The inflammation of all experimental rabbit eyes, including cornea edema and anterior chamber exudation were observed via zoom-photo slitlamp microscope 1,3,7,14,30 d postoperation. Meanwhile, aqueous humor was drawn for white blood cell (WBC) counting and classifying, and for NO-2／NO-3 and cytokine assay, including interleukin-2(IL-2), tumour necrosis factor-α(TNF-α). Statistics were taken by SPSS softwear. RESULTS: (1) Total WBC in aqueous humor were higher and anterior chamber exudation were more severe in ECCE+IOL group than that in ECCE group and control group. (2) The level of IL-2 and TNF-α and the content of NO-2／NO-3 in aqueous humor of ECCE+IOL group were higher than that in ECCE group and control group 1-14 d postoperation respectively, and it increased to peak value at 3-7 d postoperation and decreased gradually after two weeks postoperation. (3) The change regularity of IL-2, TNF-α and NO-2／NO-3 in each group were basically similar, i.e. when the level of cytokine (IL-2 and TNF-α) was normal, the content of NO-2／NO-3 was normal too, when the level of cytokine (IL-2 and TNF-α) increased, the content of NO-2／NO-3 increased too. CONCLUSION: The intraocular inflammation after ECCE+IOL was more severe than that after simple ECCE. NO, IL-2 and TNF-α play an important role in intraocular inflammation after intraocular lens implantation. The changes of IL-2 and TNF-α level was closely related with NO content in aqueous humor.
作者： 刊期： 2001年第08期
Chickens were infected with chicken anemia virus (CAV) at one-day-old and vaccinated with La Sota vaccine 8 days later. Meanwhile, uninfected chickens were vaccinated as controls. At 7, 14 and 28 days post vaccination, the content of IgG,IgM,IgA and HI titer in serum, the number of T cells, IgG, IgM and IgA antibody producing cells in thymus, bursa and spleen, the proliferative response of T、B cells, the inductive activity of interleukin 2 (IL-2) and interferon (IFN) in thymus and spleen were tested. The results showed that the content of IgG, IgM, IgA and hemoagglutination inhibition (HI) titer in serum, the number of T cells, IgG, IgM and IgA antibody producing cells in thymus, bursa and spleen, the proliferative response of T cells and B cells as well as the inductive activity of IL-2 and IFN in thymus and spleen of infected-vaccinated chickens significantly decreased compared with the control. These results indicated that the immunofunction and immunoregulation were dropped post ND vaccination of CAV-infected chickens.
作者： 刊期： 2001年第08期
This experiment, using cultured bovine pulmonary artery endothelial cells (BPAEC), was undertaken to investigate roles of endogenous ONOO- in lipopolysaccharide (LPS)-caused injury to endothelial cells. The fluorescent intensity of nitrotyrosine (NT), a marker of ONOO- generation, in BPAEC represented content of endogenous ONOO- generation. The fluorescent intensity of NT and number of NT positive cells were detected with flow cytometry, and the percentage of NT positive cells was calculated. Results were as follows. (1) LPS (1 mg/L, 5 mg/L and 10 mg/L) caused marked increase in fluorescent intensity of NT in a dose dependent manner. The number and percentage of NT positive cells were markedly increased (P＜0.05). Aminoguanidine (AG), a selective inhibitor of inducible nitric oxide synthase, inhibited the increase in fluorescent intensity of NT in BPAEC induced by LPS. However, the number and percentage of NT positive cells had tendency to reduce. (2) LPS caused the enhancement of MDA content and activity of LDH in cultured supernatant (P＜0.01). AG reversed the enhancement of MDA content induced by LPS (P＜0.01). In contrast, AG had marginal effect on activity of LDH. (3) LPS induced the increase in apoptotic rate in BPAEC in a dose dependent manner. Some BPAEC stained with fluorescent probe ethidium bromide showed morphological features of apoptosis with chromatin condensation and nuclear fragmentation. AG reduced the apoptotic rate and number of apoptotic cells, both of which were still higher than those of vehicle group (P＜0.05). (4) LPS inhibited mitochondrial respiration. Effect of LPS on mitochondrial membrane potential (ΔΨ) depended on the doses of LPS. 1 mg/L LPS led to a little increase in ΔΨ, while 5 mg/L and 10 mg/L LPS significantly reduced ΔΨ. In conclusion, LPS caused injury to cultured BPAEC and increased production of ONOO-. Cytotoxicity of LPS may be mediated by endogenous ONOO-.
作者： 刊期： 2001年第08期
The present study was undertaken to investigate the effect of human PMNs on the production of TNF-α by the human peripheral blood mononuclear cells (PBMCs) and to elucidate its tentative mechanism. Human PMNs and PBMCs were isolated from the venous blood of healthy donors by dextran sedimentation and density gradient centrifugation. In the presence of lipopolysaccharide (LPS), PMNs and PBMCs were cocultured at the ratio of 2:1 for 20 h and the concentration of TNF-α in the supernatant was measured by enzyme-linked immunosorbent assay. The binding rate of monocytes with the fluorescein isothiocyanate-labeled LPS (FITC-LPS) and the mean surface fluorescence intensity of monocytes were analyzed by flow cytometry. Results showed that PMNs were capable of inhibiting the TNF-α release from PBMCs (P＜0.05). PMNs suppressed the TNF-α release from PBMCs by 45% on average when PMNs and PBMCs cocultured at the ratio of 2:1. Paraformaldehyde-fixed PMNs still demonstrated the same inhibition (P＜0.05)，which proved that the inhibition was dependent on cell-to-cell contact and suggested that effector molecules responsible for this effect existed on the cell surface of PMNs. In the presence of PMNs, the binding rate of monocytes with the FITC-LPS and the mean surface fluorescence intensity of monocytes were not affected compared with PBMCs alone (P＞0.05). As incubation time was prolonged, the binding of FITC-LPS to monocytes increased (P＜0.05). Thus PMNs did not block the binding of LPS with monocytes. It was concluded that PMNs suppressed the TNF-α release from PBMCs via cell-to-cell interaction. In a cell-contact dependent manner, PMNs might interfere with the signal transduction pathway through which LPS activated PBMCs, thus attenuating the response of PBMCs to LPS and downregulating the TNF-α release.
作者： 刊期： 2001年第08期
AIM：To investigate the role of NF-κB in the activation of inducible nitric oxide synthase (iNOS) gene by tumor necrosis factor alpha (TNF α) and lipopolysaccharide (LPS) in endothelial cells and effect of antioxidant on the induction of iNOS. METHODS：Rat pulmonary microvascular endothelial cell (RPMEC) was cultured and the cells were identified with antiendothelial cell antibody CD31 using immunohistochemistry(ABC). The concentration of nitrite in the culture media was determined based on Griess reaction. iNOS mRNA was analyzed using RT-PCR and Northern blot. NF-κB in cell nuclei was detected with electrophoresis mobility shift assay (EMSA). RESULTS：A marked production of nitrite in RPMECs was found after 24 hours treatment with TNF α(105 U/L) and LPS (1 mg/L) (P＜0.01). The level of iNOS mRNA increased significantly after adding TNF α(105 U/L) and LPS (1 mg/L) to the cell media for 2 hours (P＜0.05). Pretreatment with cycloheximide (CHX, 10 mg/L) or antioxidant, PDTC (0.1 mmol/L) or NAC (20 mmol/L) significantly decreased nitrite production and iNOS mRNA expression induced by TNF α(105 U/L) and LPS (1 mg/L) (P＜0.05). Furthermore, there was a dose-effect relationship between PDTC/NAC and inhibitory effect. TNF α (105 U/L) and LPS (1 mg/L) triggered the activation and translocation of NF-κB. This effect was blocked by adding PDTC (0.1 mmol/L) or NAC (20 mmol/L) to the cell media for 1.5 hours.CONCLUSION：1.TNFα and LPS may induce iNOS gene expression at transcriptional or posttranscriptional level. The upregulation of iNOS depends on new protein synthesis. 2. The induction of iNOS gene expression by TNFα and LPS is dependent on the activation of NF-κB. 3. Antioxidants may inhibit the induction of iNOS gene through the inhibition of NF-κB activation.
作者： 刊期： 2001年第08期
AIM: It is well known that different injuries will result in different consequences. In this paper, we investigated the morphological change of spinal cord following crushed, hemi-sectioned and transected injury. METHODS: Sixty Wistar rats were randomly divided into 4 groups: intact group, crushed spinal cord injury group (cSCI), hem-sectioned SCI group (hSCI) and transitioned SCI group (tSCI). The models of SCI were established by the method in our laboratory. The animals in each group were sacrificed respectively at 24 hours, 7 and 24 days after operation. The L2 spinal cord which located in the caudal of injury site was taken respectively from each animal in each group and sectioned into frozen sections (20 μm). The sections were stained by hematoxylin and observed under light microscope. The number of neurons in dorsal and ventral horn was also counted. RESULTS: In cSCI group, some neurons appear to atrophy compared with that of intact group, but the number of neurons did not decrease apparently than that of intact group (P＞0.05). Comparatively, some cavities were observed in dorsal and ventral horn in hemi-sectioned and transitioned SCI group. And the number of neurons in dorsal horn and ventral horn decreased greatly at 24 hours, 7 and 21 days compared with intact group (P＜0.05). The results indicated that the decrease of neuronal number in dorsal horn and ventral horn after injury resulted from hSCI and tSCI, but not from cSCI. As a result, some different strategies should be considered for different injuries. For example, some neurotrophic factors may be useful in cSCI, but, many neurons have disappeared following hSCI and tSCI, therefore, other strategies that increase the number of neurons should be considered too. CONCLUSION: Our results provide the important morphological evidences on the change of spinal cord following cSCI, hSCI and tSCI. The data will be useful in treatment of SCI in the future.
作者： 刊期： 2001年第08期
AIM and METHODS: After the fine carbon fiber powder was injected into the right subdural space of the mice, dynamic observation was carried out on their movement and histopathological changes. RESULTS: 1-52 weeks after the injecting, no neurological changes concerning with the implanting of the carbon fiber powder were found in the experimental mice. The fine carbon fiber extensively located on the inter surface of the dura mater membrane of the right temporalis and the out surface of pie mater. Only slight inflammatory cells reaction was found under optical microscopes. The degree of inflammation reaction are Grade Ⅱ 1 week after injection and was Grade Ⅰ 2 weeks after injection, inflammation was disappeared 4 weeks after injection. No obvious fiber membrane was found around the implanted materials. No significant differences were found between the experimental and the control group.CONCLUSION: It was showed that the carbon fiber shares excellent histocompatibility after injected into the subdural space and subarechnoid cavity of the right temple of mice.
作者： 刊期： 2001年第08期
The neurofilament proteins (NFPs) from different neuronal tissues including Alzheimer and Huntington disease gray matter, rat brain gray, white matter and spinal cord were separated biochemically into two major fractions. A systematic investigation on the distribution, expression and phosphorylation of NFPs in those fractions was undertaken in the present study. It was found that only non-phosphorylated NF-H and NF-M, but not NF-L subunit were detected in Alzheimer brain gray matter high speed supernatant, whereas all neurofilament subunits including non-phosphorylated and phosphorylated were measured in high speed pellet fraction of the same tissue. The hyperphosphorylation of NF-H and NF-M in Alzheimer brain was shown by phosphorylation dependent monoclonal antibodies SMI31 and SMI34. This hyperphosphorylation was confirmed by non-phosphorylation dependent antibody SMI32 with dephosphosphorylation of the samples. Furthermore, an increased amount of NF-H, NH-M and NF-L, detected by SMI33 and NR4 respectively, was also observed in Alzheimer samples, in which the elevation in NF-L was significant. A significantly different immunoblot patterns in distribution, expression and phosphorylation were determined in various position of the neural system and alternative fractions. To our best knowledge, this is the first data shown definite abnormality of NFPs in Alzheimer disease. The information obtained in the present study will be extremely valuable in further study of the proteins both in physiological and pathological conditions.
作者： 刊期： 2001年第08期
AIM:Aquaporins (AQP) are very important for the water transport across cell membrane. There are at least 10 mammalian AQPs( aquaporins 0-9) distributed in various organs and different kinds of cells. Each AQP has a distinct organ distribution, and this distribution could be useful in presuming the biological function of the aquaporin. The aim of this study was to figure out the distribution of aquaporin 7 (AQP7) and aquaporin 8(AQP8).METHODS：Semi-quantified RT-PCR was employed in this research. The ratio of OD value of target gene products divided by which of control gene products was calculated. Among 35 organs, testis, epididymis, skin, muscle, rectum, lung, bronchus, lymph node, stomach, duodenum, jejunum, ileum, colon, pancreas, liver, gall bladder, spleen, mammary gland, uterus, placenta, tonsil, urinary bladder, thyroid came from normal area of removed samples during operation. cDNA library of Prostate, thymus, salivary gland, penis, carotiol artery, adrenal gland, occipital lobe of brain, temporal lobe of brain, frontal lobe of brain, parietal lobe of brain, mid brain, choroid plexus are purchased from OriGene biotechnique company.RESULTS：①AQP 7 mRNA was found in testis, muscle, gall bladder, carotiol artery, lymph node and adrenal gland, and maximum expression of AQP 7 was in testis.②AQP 8 mRNA was found in pancreas, testis, skin and colon. and maximum expression of AQP 8 was in pancreas.CONCLUSION:Coexistence of AQP 7 and 8 in testis was confirmed, which suggested that both of these two aquaporins were involved in the regulation of testis function.
作者： 刊期： 2001年第08期
Our previous experiments confirmed that endothelial derived ONOO- mediated injury to cultured BPAEC induced by LPS, and that CCK protected endothelial functions against the detrimental effect of LPS in vitro. In the present study, using cultured BPAEC, we investigated effect of CCK on LPS-induced generation of ONOO- in BPAEC and on injury to BPAEC induced by LPS. Results were as follows.(1)CCK inhibited increase in endothelial generation of ONOO- induced by LPS, for fluorescent intensity of NT in BPAEC reduced from (6.55±0.30) AU of LPS group to (4.37±0.08) AU (P＜0.01), the latter being still higher than (3.27±0.15) AU of vehicle group (P＜0.01). In contrast, the number and percentage of NT positive cells reduced a little. Proglumide, a nonspecific inhibitor of CCK receptors, might in part reverse the effect of CCK. (2)CCK markedly reduced MDA content in supernatant in LPS group (P＜0.01), which was completely reversed by proglumide(P＜0.01). Activities of LDH in supernatant in LPS, LPS+CCK group were (85.30±8.66) U/L and (71.33±4.07) U/L, respectively. Proglumide elicited increase in activity of LDH ［(81.00±6.35) U/L］. However, effect of CCK or proglumide was not significant in the alterations of activity of LDH. (3)CCK significantly inhibited increase in apoptotic rate in BPAEC induced by LPS, from 13.50%±0.60% of LPS group to 5.35%±0.25%(P＜0.001), the latter being completely reversed by proglumide with apoptotic rate of 11.45%±0.65%(P＜0.05).These results further confirmed that CCK afforded the cyoprotection for the mitigation lipoperoxide damage and inhibition of apoptosis in BPAEC induced by LPS. The cytoprotection of CCK may be mediated by CCK receptors and related to the reductive ability of endothelia to generate ONOO- induced by LPS.
作者： 刊期： 2001年第08期
It is well known that the main brain lesion in Alzheimer's disease (AD) brain is neurofibrillary tangles (NFT) and senile plaques (SP). The amount of NFT is positively correlated with clinical degree of dementia in AD. It is also well studied that the major component of NFT is abnormally hyperphosphorylated microtubule associated protein tau that is caused by an imbalance of protein kinase and protein phosphatase (PP). To reconstitute a specific AD model based on the above hypothesis, we have injected separately calcium calmodulin dependent protein kinase (CaMKKII) activator, bradykinin and PP-2B inhibitor, cyclosporin A into rat hippocampus in the present study. The results showed that the injection of bradykinin caused learning and memory deficient in rats as well as Alzheimer-like tau phosphorylation, including Ser-262/356, Thr-231/235 and Ser-396/404. On the other hand, the injection of cyclosporin A induced the same phosphorylation sites as above except Ser-262/356, however, it did not mimic rat behavior abnormality as bradykinin injection did. The data suggested that activating of CaMKII and the phosphorylation of Ser-262/356 at tau might responsible for the lesion of learning and memory in our model rats. We also incubated PP-2A and PP-1 inhibitor, okadaic acid with human neuroblastoma cell line (SH-SY5Y), and found that (1) inhibition of above PPs induced Alzheimer-like phosphorylation and accumulation of neurofilaments, and Alzheimer-like microtubule disruption, (2) melatonin showed certain protection of the cell from okadaic acid toxicity. The data obtained from this study is significant in AD specific model study.
作者： 刊期： 2001年第08期
The present study aimed to examine roles of histamine neurons in leptin signaling pathways and leptin resistant states. H1-receptor knockout (H1KO) mice showed no change in daily food intake, adiposity, growth curve, basal expression of hypothalamic neuropeptides, uncoupling proteins (UCPs) or ob gene. However, H1KO mice fed with high fat diet increased fat deposition and ob gene expression more excessively. Leptin-induced feeding suppression was attenuated in H1KO mice. Chronic leptin treatment decreased visceral fat and up-regulated UCPs expression in brown and white fat. These effects of leptin were attenuated in pair-fed H1KO mice. Chronic histamine or histidine treatment decreased body weight, body fat deposition, and serum glucose and insulin in diet-induced obese, ob/ob and db/db mice. Activation of histamine neurons suppressed ob gene expression in the fat tissue together with elevation of seurm leptin and UCPs mRNA. These actions of neuronal histamine were attenuated in the double knockout mice, i.e., db/db mice with H1KO. Taken together, H1KO mice, a novel leptin resistant model, elucidate essential roles of H1-R in energy intake and expenditure as a down-stream-signaling message of leptin actions in the brain. The anti-obesity and anti-diabetic effects of histamine neurons provide a new insight into therapeutic strategies on human obesity and diabetes with leptin resistance.
作者： 刊期： 2001年第08期
AIM： To investigate the efficiency of cytosine deaminase adenoviral/5-fluorocytosine system on prostate cancer cell lines. METHODS: Cell culture, infectivity test and sensitivity test, observing the bystander effect and animal model experiment were carried out. RESULTS: All the established prostate cancer cell lines were eventually infectable, but ratio of vector/cell and time of exposed at which infection occurs was dependent on the cell lines. The expression of transfered cytosine deaminase gene peaked at different days, but persisted beyond 11 days. The prostate cell lines were sensitized to the 5-fluorocytosine by infection with the cytosine deaminase gene adenoviral vector, and only 5% of the LNCap and 10% of the RM-1 cells infected were required for 100% cell death. In the animal model, there was significant eradiation of tumor growth at the ratio of 400 vector particles/cell and with the systematic treatment of 5-fluorocytosine. CONCLUSION: The adenoviral vector carrying a cytosine deaminase transcription unit can sensitize the prostate cancer cell lines to 5-fluorocytosine, and the system can significantly inhibit the growth of prostatic tumor in mice.
作者： 刊期： 2001年第08期
Cytokines are involved in both host defense and inflammatory lung injury. Recent work from our laboratory and others has demonstrated that in addition to classical immune cells, lung alveolar epithelial cells (or pneumocytes) can also produce cytokines in response to various stimuli. This new knowledge has advanced our view of the host defense system in the lung. The regulatory mechanisms of cytokine production have been studied in great detail at various cellular and molecular levels, but the mechanisms of intracellular cytokine transport are largely unknown. Our recent studies suggest that the cytoskeleton could play an important role in mediating intracellular cytokine trafficking. This could be an important regulatory step for cytokine production. For example, lipopolyssacharide (LPS) induced tumor necrosis factor-α (TNF-α) from rat pneumocytes, which was further enhanced by a microfilament-disrupting agent. LPS also induced macrophage inflammatory protein-2(MIP-2), a chemokine for neutrophil recruitment and activation, from rat pneumocytes. This effect was enhanced by microtubule-disrupting agents. We speculate that both microfilaments and microtubules are involved in regulating cytokine transportation in pneumocytes through different mechanisms. Further investigation in on going in my laboratory. From a clinical perspective, if we understand the mechanisms regulating cytokine production and release from lung alveolar epithelial cells, we may be able to enhance or inhibit release of crucial cytokines depending on the clinical situation.
作者： 刊期： 2001年第08期
In this study we found: 1, There was endogenous ONOO- formation in lungs in the early stage of endotoxic shock. Exogenous ONOO- led to increase in microvascular permeability, severe lung pathological changes and enhanced MDA content. 2, It was, for the first time, found that responses of isolated pulmonary artery preincubated with ONOO- showed abnormal manifestations. (1) Low dose of ONOO- let to the inhibition of endothelial dependent relaxation, but enhacement of contractile response, both of which were similar to changes of reactivity in isolated pulmonary artery induced by LPS. (2) High dose of ONOO- reduced contractile response to PE and relaxation to SNP. 3, ONOO- had direct effect for relaxation of precontracted isolated pulmonary artery. The relaxing action of ONOO- was weak and was negtively regulated by endothelial cells, supporting the notion that ONOO- may be involved in pulmonary hypertension in the early stage of endotoxic shock. 4, It was, for the first time, found that LPS-induced increase in endogenous ONOO- generation in BPAEC and that endogenous ONOO- mediated injury to BPAEC induced by LPS, which may be a novel mechanism for endotoxin-elicited damage to endothelial cells. 5, Exposure of pulmonary artery to LPS led to reduction in endothelial dependent relaxation but enhancement in contractile response, both of which were reversed by concomitant exposure to CCK and LPS. 6, CCK protected cultured BPAEC against the detrimental effects of LPS such as lipoperoxide damages and cellular apoptosis as well as LPS-induced endogenous ONOO- formation. The underlying mechanism of CCK for cytoprotection may be mediated by its receptors and related to its reduced ability of endothelia to generate ONOO- induced by LPS.
作者： 刊期： 2001年第08期
AIM: Expression of bcl-2 in Marek's disease (MD) tumor was analyzed and relationship between bcl-2 gene and lymphomagensis clarified. METHODS: Model of MD tumor was reproduced on SPF White Leghorn chickens, collected tumor and normal tissues, conserved in liquid nitrogen. Total RNA in the tissues was obtained by AGPC method, detected their concentration and analyzed by 1% denatured agrose gel containing formaldehyde. A plasmid (pTTCB1) that contains chicken bcl-2 1.2 kb fragment was amplified, extracted and digested with restriction endonuclease (RE), the 1.2 kb fragment was recoveried and puarified with purigene kit, labeled probes with [α-32P]. Expression of bcl-2 was analyzed with Dot blot and Northern blot hybridization. RESULTS: (1) There are bcl-2 transcripts in MD tumor and relevent normal tissues; (2) Expression of bcl-2 in MD tumor is much higher than that in normal tissues. CONCLUSION: Our findings indicated that expressive products of chicken bcl-2 promotes lymphomagenesis of MD by blocking apoptotic cell death.
作者： 刊期： 2001年第08期
Plasmin/plasminogen activators (PA) are the serine enzyme which digests fibrin and/or fibrinogen. Plasmin is produced by the cleavage of its precursor, plasminogen by PAs (urokinase-type PA and tissue-type PA). These events are expected in the thrmbolytic therapy for thromboembolic deseases. Apart from the blood fibrinolysis mentioned above, new role of plasmin/plasminogen activators has been extensively investigated in the field of cellular biology. On the cell surface, the receptor for urokinase-type PA (u-PAR) was found (that for t-PA has not cloned yet). Then, plasmin as well as u-PA itself activates pro-form of matrix metalloproteinases (MMPs) around the pericellular space. These proteolytic activities by u-PA, plasmin and MMPs induce the degradation of extracellular matrix (ECM), affording the cells certain enviroment for their biological function. Further, the coupling of u-PA/u-PAR system and integrins can generate intracellular signal transductions which take part in the regulation of cell proliferation, attachment or migration followed by various physiological and pathophysiological functions. These serial mechanisms are the principle of pericellular proteolytic cascade.
作者： 刊期： 2001年第08期
Bradykinin (BK) is a calcium/calmodulin dependent protein kinase Ⅱ (CaMKⅡ) specific activator, and Cyclosporin A (CSA) is reported to suppress protein phosphotase (PP)-2B activity. In vitro studies have shown that CaMKⅡ and PP-2B play an important role in Alzheimer-like phosphorylation of microtube-associated protein tau. To reconstitute an animal model based on the imbalance of protein kinase (s) and protein phosphatase (s) seen in Alzheimer brain, we injected BK and/or CSA into rat hippocampus. The results from behavioral study showed that an obvious disturbance in learning and memory was seen with BK or BK plus CSA injected rats. Moreover, the behavior abnormality appeared earlier in aged rats than young adults of the same kind after the injection. On the other hand, no obvious dysfunction in living and behavior was observed with CSA alone injected rats. The results obtained by immunohistochemical assay indicated that the staining for M4, 12E8, PHF-1 and CaMKⅡ was stronger, and for Tau-1 was weaker in BK injected rats compared with Control group. It was also found that the binding of M4 and PHF-1 but not 12E8 to tau was significantly increased in CSA injected rats. As the same as BK injection, binding of Tau-1 to tau was decreased after CSA injection. The immunostaining for 12E8,PHF-1 and CaMKⅡ was increased, whereas for Tau-1, M4, and GSK-3 was decreased after combination injection of BK and CSA. In addition, the staining of PP-2B decreased in all the three models. To our knowledge, this is the first data shown in vivo that the activation of CaMKⅡ induces both Alzheimer-like tau phosphorylation and behavioral disturbance.
作者： 刊期： 2001年第08期
AIM: To determine the evolutionary pattern of parenchymal cell apoptosis in multiple organs early after intestinal ischemia-reperfusion(I/R) and its induction mechanisms and the role of apoptosis in triggering SIRS/MODS. METHODS: An I/R model was reproduced by clipping and releasing the superior mesenteric artery in rats and mice. Flow cytometry, electron microscope, DNA agarose gel electrophoresis, TUNEL method, fluorescent and Gomori's silver-HE staining were used to detect apoptosis. Distribution features of apoptotic parenchymal cells in multiple organs were observed. Immunohistochemical staining of HSP 70 and Bcl-2 were performd to study the induction mechanisms of apoptosis.RESULTS and CONCLUSION: 1. Damage of the liver, lung, gut and kidney was appeared in early phase of I/R. The percentages of apoptosis in parenchyma organs increased progressively. The percentages of cell necrosis increased with the prolonged I/R duration. 2. Percentages of apoptosis were much higher near the central veins of liver lobules, in the outer medulla of the kidney, and the antimescenteric border of intestinal mucosal epithelium because of ischemia. 3. The expression of HSP 70 increased and Bcl-2 reduced in the areas mentioned above because of hypoperfusion. 4. Apoptosis of I/R hepatocytes, splenocytes and thymocytes was obviously increased after Kupffer cell blockage with GdCl3, showing the functional state of Kupffer cells may play an important role in SIRS/MODS.
作者： 刊期： 2001年第08期
The primary action of corticotropin releasing hormone (CRH) is stimulation of the synthesis and release of adrenocorticotropic hormone and β-endorphin from the pituitary in response to stress. In addition, a number of studies indicate that CRH exerts other physiological actions within the central nervous system which are independent of the pituitary. These include increased body temperature and thermogenesis. However, the intracellular mechanism responsible for pyrogenic action of CRH is still unclear. The purpose of these studies was to determine whether or not cAMP was involved in the pyrogenic action of CRH in the rat. Intracerebroventricular (icv) microinjection of CRH (2.5 μg, 5.0 μg, 10 μg) caused increases in colonic temperature and hypothalamus cAMP level in conscious rats. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (icv) of an adenylate cyclase inhibitor, 2，，3，-dideoxyadenosine (DDA, 30 μg) or an inhibitor of cAMP-dependent protein kinase, adenosine-3，，5，-(cyclic) monophosphorothionate (Rp-cAMPs, 15 μg). This is the first report demonstrating the pyrogenic effcet of centrally administration of CRH on the rat via the cAMP-mediated protein kinase signal transduction pathway.
作者： 刊期： 2001年第08期
Orexins, hypothalamic neuropeptieds, are involved in modulation of food intake and arousal state. To examine further physiological roles of orexin in brain function, the effects of centrally administered orexin- A on body temperature was investigated in rats. Assessed by a telemetry-sensor system implanted into the abdominal cavity, infusion of orexin-A into the third cerebroventricle increased body temperature in a dose-responsive manner. Cumulative ambulatory activity was concomitantly increased during 6 h but not 12 h after administration of orexin-A. Expression of uncoupling protein 1 (UCP1) mRNA in brown adipose tissue, as a marker for peripheal thermogenesis which affects body temperature, failed to increase after orexin-A administration. Expression of UCP3 mRNA in skeletal muscle but not UCP 2 in white adipose tissue was upregulated by infusion of orexin-A. The resulting information indicates that orexin neuron regulates body temperature in coordination with control of arousal system independently of peripheral thermogenesis through the BAT UCP1.
作者： 刊期： 2001年第08期
To study the direct effect of E.Coli endotoxin on the production of nitric oxide by endothelial cells, the second passage of cultured human umbilical cells was stimulated by serial doses of endotoxin (1 g/L, 10 mg/L, 100 μg/L, 10 μg/L, 1 μg/L), and the content of nitric oxide in supematant of culture and the viability of endothelial cells 6 hours after the stimulation were obcerved. The result showed that endotoxin had a slightly inhibitory effect on both the production of nitric oxide and the viability of endothelial cells at low doses (1 μg/L, 10 μg/L, 100 μg/L), especially the dose of 100 μg/L ［(608.63±11.64) μmol/L, versus that of unstimulated grouop (629.46±13.36) μmol/L, P＜0.05］. While the high doses of endotoxin exerted a big increasing in production of nitric oxide and a big decrease in the viability of endothelial cells, especially the dose of 1 g/L (NO: 722.58 μmol/L±32.18 μmol/L, versus that of unstimulated group P＜0.01; viability: 73.63%±8.50%, versus that of unstimulated group, P＜0.01). These could be concluded that low doses of endotoxin mainly resulted in functional changes in endothelial cells, such as decrease in relaxing factor (nitrc oxide), while high doses endotoxin exerted lethal effects on endothelial cells accompanied with high production of nitric oxide, which might be related to the death of cells.
作者： 刊期： 2001年第08期
Elemene is a new anticancer drug isolated from a Chinese traditional medicine Curcuma aromatica. In previous work, we discovered that tumor cell vaccine (TCV) treated with oleum Curcuma aromatica or elemene could induce significant immunoprophylactic effect against a variety of aminal tumor strains and the method of preparation of elemene combo-TCV(EC-TCV) already got China's inventive patent. In this paper we further studied the active immunotherapeutic effect and the possible cellular/molecular mechanisms of EC-TCV immunization. The results were as follows:(1) EC-TCV immunization showed significant therapeutic effects (P＜0.05) against murine Ca761 syngeneic mammary carcinoma (H-2k) and HCa-F allogeneic hepatic carcinoma (H-2-) models; (2) The spleen cells of Hca-F EC-TCV immunized mice displayed higher cytotoxicity and IL-12 level while the secretion of IL-10 was decreased (P＜0.05); (3) Similar to heat shock, elemene(E), mitomycin C(MMC) and glutaraldehyde (G) could act alone as stressor, and induce significant changes of the expression of membrane heat shock proteins(HSP70 or/and HSP90) on L615 leukemia and HCa-F hepatoma cells and the EC-TCVs (E+MMC+G treated in combination) showed the highest level of membrane HSPs expression (P＜0.05 or P＜0.01 );(4) The HSP70-peptide complex isolated from HCa-F EC-TCV through ADP-agarose affinity chromatographic system could induce active immunoprotection against lethal dose challenge of HCa-F hepatic cancer cell but could not protect against the cross challenge of lethal dose of L615 leukemia. The results indicated that the immunoprotective effect of EC-TCV was in some extent tumor-specific, MHC-nonrestricted, and HSPs might play an important role in its molecular mechanisms.
作者： 刊期： 2001年第08期
In this study, immunotoxin (IT) was prepared by conjugating BAC5 and CT with SPDP. The effects of IT on NPC and its mechanisms were explored using double labeled with radioactive nuclides, immunography and electron microscope technique in vivo and in vitro. The specific concentration of BAC5 in the tumor area showed. The radioactivity rate of tumor/nontumor (T/NT) was up to 10.26. IT had cytotoxic effects both on the cultured CNE-2 cell line and tumor multicell spheroides. In vivo, the preliminary result indicated that IT also had a inhibitory action on the nude mice models bearing human NPC (Reported in another article). Under electron microscope, the necrosis and apoptosis of tumor cells were found. The membranes of most tumor cells were found intacted not or corrosined, some of them had the character of apoptosis, including reduce of tumor cells membrane villi, condensation of cytoplasm and pyknosis or cleavage of nuclear. There were many of apoptosis bodies, which were occasionally phagocytosed by tumor cells. The infiltration of immunocytoes in tumor tissue could be seen. The results indicated that BAC5 can specifically combine with NPC cells and BAC5-CT has the inhibitory effect on NPC in vitro and in vivo, mechanism of which may be related to the effects that ‘warhead' CT dissolute the membrane of tumor cells directly, or/and IT promote the infiltration of immunocytoes so as to induce the apoptosis of tumor cell.
作者： 刊期： 2001年第08期
AIM and METHODS: To investigate the effect of endotoxin on the celluar activity and secretion of endothelin-1 by radioimmunoassay and MTT methods in cultured human umbilical vein endothelial cells stimulated by E coli endotoxin (E coli O55:B5, Sigma) of various concentrations (1 g/L, 100 mg/L， 10 mg/L，1 mg/L，100 μg/L，10 μg/L, 1 μg/L) and at the same time interval (HUVEC stimulated by endotoxin for 6 hours) in vitro.RESULTS:Endotoxin showed a slightly inhibitory effect on the viability of endothelial cells at low doses (1 μg/L， 10 μg/L, 100 μg/L, 1 mg/L). The viabilities were 92.00%±1.45%， 91.81%±2.03%, 89.52%±1.49%, 88.35%±1.88%, respectively, versus control group, P＜0.01. The cells were impaired significantly at the higher dose of LPS (100 mg/L), the viability was 80.49%±8.76%, versus control group, P＜0.01. The cells were killed evidently at the concentration of LPS (1 g/L), the viability was 73%±8%, versus control group, P＜0.01. The secretion of ET-1 increased gradually with the concentration of endotoxin manifolding. The concentration of ET-1 reached its peak at the dose of 100 μg/L, and it was (324.384±17.023) ng/L, versus control group (251.636±17.023) ng/L, P＜0.01. Endotoxin was effective in stimulating the endothelial cells to secret ET-1 in a dose dependent manner. CONCLUSION: These findings suggested ET-1 may be one of the important factors in endotoxic shock, and the increase in plasma ET-1 level in endotoxemia may be associated with increase in ET-1 secretion.
作者： 刊期： 2001年第08期
AIM: To investigate the role of catecholamine, angiotensin converting enzyme(ACE) and adenosine triphosphatase in ischemic preconditioning in isolated rat hearts. METHODS: Isolated perfused rat heart was subjected to global ischemia for 40 min followed by reperfusion for 10 min (I/R). Preconditioning (PC) was induced by 5 min of ischemia and 10 min of reperfusion. The tissue concentrations in NE, and ACE, ATPase activities were determined in left ventricle in the PC and I/R groups by fluorometry and spectrophotometry. RESULTS: There were no significant difference in NE and ACE between PC and I/R groups. PC hearts showed improved recovery of the contractile function after 40 min ischemia/10 min reperfusion, but activities of the myocardial total ATPase, Mg2＋-ATPase, Na＋K＋-ATPase were inhibited markedly compared with I/R group. CONCLUSION: The inhibited myocardial ATPase may be involved in the mechanism of ischemic preconditioning protection in the isolated rat heart. Endogenous myocardial norepinephrine and ACE activation are not essential for ischemic preconditioning in the isolated rat heart.
作者： 刊期： 2001年第08期
Antibodies to glutamic acid decarboxylase (GADab) is considered to be a marker for the autoimmune process against pancreatic β cells. Indeed, nearly 70% of patients with type 1 diabetes is repoted to be GAD ab＋. A subgroup of patients diagnosed as type 2 diabetes has GADab. Therefore, it is questioned whether GADab＋ patients with type 2 diabetes represent a late onset of type 1 diabetes or a unique disease entity. Fifty five GADab＋ patients with type 2 diabetes were compared with 137 GADab- patients. They were admitted to Abe Diabetes Clinic for the control of diabetes. The age at onset of these patients was ＞30 years, and did not require insulin therapy for at least 6 months from the disease onset. The GADab＋ patients had lower urinary C-peptide concentrations (uCPR) ［(47.8±48.9) μg/d vs (58.1±49.9) μg/d, P=0.034］ than the GADab- patients. The GAD＋ patients were assigned insulin therapy more often (81.8% vs 56.3%, P=0.0038) and earlier ［(8.5±7.5)years vs (10.1±7.3) years, P=3.3×10-12］ than the GAD- patients. The levels of uCPR were associated with the titers of GADab (r=0.32, P=0.038). Among the susceptible HLA alleles for type 1 diabetes, the frequencies of B54 and DRB1* 0405 alleles, but not B61 and DRB1*0901 alleles, were increased among GADab＋ patients. There data suggest that the GADab＋ type 2 diabetes has an autoimmune nature, although the extent of insulin dependency and the distribution of HLA susceptible alleles are different from type 1 diabetes.
作者： 刊期： 2001年第08期
AIM：The mechanism of tumor necrosis factor-alpha (TNF-α) induced pulmonary artery hypertension(PAH) in endotoxic shock (ES) is not clear. Cholecystokinin-octapeptide (CCK-8) had anti-ES and anti-PAH effects. The impact of CCK-8 on changes in vascular reactivity and endothelial ultrastructure induced by TNF-α was studied. The role of nitric oxide (NO) was preliminarily studied. METHODS：Rabbit pulmonary artery rings were divided into four groups: TNF-α, TNF-α+CCK-8, CCK-8 and Vehicle. The rings were incubated for 2 h, 7 h or 14 h. Relaxative responses to ACh(10-8-10-5 mol/L)， SNP (10-9-10-6 mol/L) and contractile responses to PE(10-8-10-5 mol/L) were generated seperately. The NOS activity of rings was detected and the ultrastructure of endothelium was observed in the groups that incubated for 7 h.RESULTS:The relaxative responses to ACh were not affected by TNF-α and CCK-8 after incubation for 2 h. TNF-α(7 h,14 h) significantly reduced ACh-induced endothelium-dependent relaxation response of pulmonary artery. CCK-8 reversed the effect. CCK-8 itself had no effect on responses of normal pulmonary artery. Contraction to PE and relaxation to SNP were unaffected by TNF-α, CCK-8. The NOS activity increased in the TNF-α and the TNF-α+CCK-8 groups. While no significant difference was obseved between the Vehicle and the CCK-8 groups. Endothelial injury in TNF-α group and alleviated changes in TNF-α+CCK-8 group were observed. CONCLUSION:TNF-α significantly inhibits endothelium-dependent relaxation, which be one of the mechanisms of PAH induced by TNF-α during ES. It was found for the first time that CCK-8 reversed TNF-α induced impairment of endothelium-dependent relaxation and alleviated structural injury of endothelium, which might be one of the mechanisms of anti-PAH effect by CCK-8 during ES. The effects of TNF-α and CCK-8 might be related to NO.
作者： 刊期： 2001年第08期
AIM: To study the expression of c-fos mRNA in brain following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expression following percussion. METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury group. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa). The injury groups were then subdivided into 5 min, 15 min, 30 min, 1h, 2h groups according to the time elapsed after injury. The expression of c-fos mRNA was studied with reverse transcription polymerase chain reaction (RT-PCR) semi-quantitatively.RESULTS: At 5 min after percussion, the induction of c-fos mRNA was increased, and remained elevated up to 2 h after brain injury.CONCLUSION: The induction and expression of the c-fos mRNA in cortex and brain stem after fluid percussion brain injury were increased rapidly.
作者： 刊期： 2001年第08期
AIM: To study the changes in PaO2 and pH in experimental hepatocarcinoma and their significance.METHODS:Qualitative analysis of the PaO2 and pH was made by three channel tissue measuring equipment in experimental hepatocarcinoma and comparative liver tissue. RESULTS:The results showed that the PaO2 in tissue of test group was significantly higher than that of control group (P＜0.01), but the pH of tissue in test group was slight higher than that of control group (P＞0.05).CONCLUSION: PaO2 might play an impontant in genesis and developmant of hepatocarcinoma. PaO2 may be helpful in diagnosis and differentiated diagnosis for hepatocarcinoma.
作者： 刊期： 2001年第08期
AIM: To study the expression of c-jun in brain stem following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expressions following percussion.METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa), and then were subdivided into 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 12 h groups according to the time elapsed after injury. The expression of c-jun was studied by immunohistochemistry and in situ hybridization. RESULTS: After percussion for 15 min, Jun positive neurons increased in brain stem progressively, and peaked at 12h. At 5min after percussion, the induction of c-jun mRNA was increased, and remained elevated up to 1h-2h after brain injury. CONCLUSION: The induction and expression of the c-jun in brain stem after fluid percussion brain injury were increased rapidly and lasted for a long time.
作者： 刊期： 2001年第08期
AIM: To study the apoptotic effect of TNFα on rat pulmonary microvascular endothelial cells (PMVEC) and the influences of Fas, NF-κB in its mechanism. METHODS: Apoptosis of PMVEC was analyzed and quantitated with TUNEL, flow cytometer. The distribution of NF-κB was detected via histoimmunochemical staining in TNF-treated cells and the control. Northern blot was applied to assess the influence of TNF on PMVEC Fas expression. Fas antibody was used to investigate the apoptotic effect of Fas on PMVEC. Activation of caspase-8 was detected with Western blot. Expression of caspase-3 was analyzed with histoimmunochemical staining. RESULTS: After treatment with 5×108 U/L TNF for 24 hours, viable PMVEC significantly diminished. Apoptosis rate was 14.0%±3.1% detected with TUNEL, and 13.1% with flow cytometer. Histoimmunochemical staining showed that NF-κB relocated from cytoplasm to the nuclear. When the cells were co-cultured with TNF and APDC, an NF-κB inhibitor, less cells were viable and more cells were positively stained with TUNEL. Fas expression in PMVEC was elevated treated with TNF. Apoptosis in PMVEC was found aggravated, when the cells were co-cultured with TNF and anti-Fas antibody. The positive rate was 24.1%±1.5% with TUNEL. Increase of caspase-8 activation was manifested by Western blot following TNF stimulation. Caspase-3 expression was found elevated using histoimmunochemical staining. Cell permeable caspase-3 inhibitor significantly ameliorated PMVEC apoptosis induced by TNF. CONCLUSION: 1. Large dose of TNF(5×108 U/L) can induce apoptosis in rat PMVEC. 2. NF-κB has a protective effect on PMVEC apoptosis. 3. TNF up-regulates Fas expression in PMVEC. And the latter takes a part in apoptosis. 4. TNF induced caspase-8 activation in PMVEC, and more caspase-3 was expressed. These may be involved in PMVEC apoptosis induced by TNF.
作者： 刊期： 2001年第08期
We have found that leptin, at physiological concentrations of 10-12 mol/L, facilitates learning and memory and LTP maintenance in Wistar rats. To explore the role of leptin recepors in learning, memory and synaptic plasticity, experiments were carried out using Zucker rats (Z), db/db mice (db), and ob/ob mice(ob). The former two have defects in leptin receptors and the latter cannot produce normal leptin. Unlike the effects observed in normal rats, high or low frequency stimulation of Schaffer collateral-CA1 synapses in hippocampal slices prepared from Z, db and ob animals failed to induce the learning and memory relevant long-term potentiation or depression in CA1 neurons. However, LTP in ob CA1 synapses was facilitated by leptin at 10-12 mol/L concentration. Moreover, the paired-pulse facilitation of CA1 synaptic potentials and intracellularly recorded postsynaptic responses to the neurotransmitters AMPA, NMDA and GABA, applied electrophoretically to the apical dendrites of CA1 neurons, were approximately the same compared to the control lean animals. In addition, unlike the second messenger responses observed in Wistar rats, calmodulin kinase Ⅱ activity in the CA1 area of Z and db animals was not activated after tetanic stimulation of the Schaffer collaterals. It has been shown that all three strains, Z, db and ob display impaired spatial learning and memory when tested in the Morris water maze. The results of these experiments indicate a close relationship between spatial learning and memory, facilitation of LTP, and calmodulin kinase Ⅱ activity.
作者： 刊期： 2001年第08期
We aimed to clarify the effects of troglitazone to prevent the development of fatty liver in Otsuka Long-Evans Tokushima fatty (OLETF) rats, an obese type2 diabetes model. Treatment of 0.2% troglitazone for 16 weeks significantly decreased blood glucose, serum insulin and free fatty acid (FFA) concentration (P＜0.05 for each) in the OLETF rats leaving their food intake, body weight and general fat pad weight unaffected compared with those of their lean littermates, Long-Evans Tokushima Otsuka (LETO) rats. Troglitazone restored increase in liver weight and hepatic triglyceride(TG) content of the OLETF rats (P＜0.05 for each). These findings were accertained by histological examination which showed that fat deposition, necrosis and vacuolization of hepatocyte, markers for fatty liver, were diminished by troglitazone treatment. Messenger RNA expression of microsomal triglyceride transfer protein in the liver, a marker for hepatic TG output, was not significantly different between OLETF and LETO rats with or without troglitazone treatment. Fatty acid composition such as C18∶1(oleic acid) showed no remarkable change after troglitazone treatment in OLETF rats, indicating no dietary influence to hepatic lipogenesis. These indicate that reduction in circulating FFA level may be one of the main mechanisms for troglitazone to prevent the development of insulin resistance and fatty liver.
作者： 刊期： 2001年第08期
To observe the effects of diabetogenic (high fat high sucrose, lacking choleserol) diet on atherogenesis in New Zealand white rabbits. Two groups of New Zealand white rabbits received regular rabbit chow (the normal control), or high fat high sucrose diet for 4 months. The levels of plasma total cholesterol, HDL cholesterol, triglycerides, insulin, and glucose were investigated, the areas of fatty streak of the aortae were measured after staining with Sodan IV, and the aortic, coronary specimens were observed with light and electron microscopies. The plasma glucose, triglycerides, and total cholesterol were increased significantly by high fat high sucrose feeding. At the end of 4 months, the early charateristics of atherosclerosis were present in the animals' vascular specimens. Our findings suggest that high fat high sucrose feeding can induce hyperglycemia, hypertriglyceridemia and atherosclerosis in New Zealand white rabbits, and this could be a potential animal model for studying the mechanisms of diabetes-accelerated atherosclerosis. This study raised a question: What is the mechanism by which high fat high sucrose feeding induces atherosclerosis?. The related hypothesis was given in this article.
作者： 刊期： 2001年第08期
Alternative splicing of the human glucocorticoid receptor (hGR) primary transcript produces two receptor isoforms, termed hGRα and hGRβ. hGRα is a ligand-activated transcription factor which, in the hormone-bound state, modulates the expression of glucocorticoid-responsive genes by binding to specific glucocorticoid response element (GRE) DNA sequences. In contrast, hGRβ dose not bind glucocorticoids and is transcriptionally inactive. We demonstrate here that hGRβ inhibits the hormone-induced, hGRα-mediated stimulations of gene expression, including glucocorticoid-responsive reporter gene (cat) and endogenous p21 gene. We also demonstrate that hGRβ can inhibit hGRα-mediated regulation of proliferation and differentiation of a human osteosarcoma cell line (HOS-8603). Our studies on the expression of hGR mRNA in nephrotic syndrome patients indicate that the hGRα/hGRβ mRNA ratio in peripheral white blood cell of hormone-resistant patients is lower than that of hormone-sensitive patients and health volunteers. These results indicate that hGRβ may be a physiologically and pathophysiologically relevant endogenous inhibitor of hGRα
作者： 刊期： 2001年第08期
AIM:To observe the antineoplastic effect of Naja Naja atra venom (NNAV) on S180 bearing mice and to study the possible antineoplastic mechanism. METHODS:We observed the effect of NNAV on tumor weight、plasma nitric oxide content、plasma endothelin content and spleen index in S180 bearing mice with different concentration and different period by means of injecting into abdomen. RESULTS:Treatment with NNAV solution of different concentration could markedly inhibit S180 growth (especially in the low concentration group and by long period) and rate of inhibiting ranged from 21 63% to 49.25%; the plasma nitric oxide content, the plasma endothelin content and NO/ET ratio in tumor bearing mice were obviously higher than those of the normal control group, while after treatment with NNAV solution, the plasma nitric oxide level, the plasma endothelin level and NO/ET ratio could be reduced markedly, and it was noticed that NO/ET ratio in the group with highest inhibiting rate was most close to that of the normal control group. The spleen index was obviously increased after treatment with NNAV solution.CONCLUSION:The antineoplastic effect of NNAV on S180 bearing mice is best in long period by means of injecting into abdomen with low concentration. The mechanism of the antineoplastic effect of NNAV may be related to lowering the plasma nitric oxide and endothelin level, regulating the NO/ET ratio and enhancing the immune response.
作者： 刊期： 2001年第08期
AIM: Ulinastatin has been reported to be beneficial for maintenance of steroid-refractory inflammatory bowel disease (IBD), but the mechanism underlying remains uncertain. Leukocyte recruitment to inflammatory site plays an important role in the pathogenesis of IBD, analysis of leukocyte and endothelium interaction may provide new avenues for treatment of IBD. In this study, we evaluated the efficacy of Ulinastatin in dextran sulfate sodium (DSS) induced colitis rat model using intravital video microscopy. METHODS: Rats were given drinking water containing 3.5% (W/V) DSS for 10 days then 1% for 14 days. DSS induced colitis rats were treated Ulinastatin 3 000 unit*kg-1*d-1 via intraperitoneum during 1% DSS feeding. Controls received distilled water for 24 days. Body weight was determined for all groups. Colitis severity was assessed using histological scoring systems by H&E sections. Intravital microscopic techniques were used to quantitate leukocyte adhesion (LA), leukocyte emigration (LE) and venular protein leakage (VPL) in rat mesentery. RESULTS: DSS induced loss of body weight, whereas Ulinastatin-treated rat showed a significant increase in body weight. Histological analysis revealed improvement of colitis such as leukocyte infiltration, loss of goblet cells, transmural edema. DSS intake elicited increase in LA, LE, and VPL compared to control group. Ulinstatin significantly reversed the increase in LA, LE, and VPL induced by DSS. CONCLUSION: Administration of Ulinastatin effectively ameliorates experimental colitis by interfering with leukocyte recruitment, and may become a potential candidate for control of inflammation of IBD.
作者： 刊期： 2001年第08期
Effects of hydrogen peroxide (H2O2)on Cryptosporidium parvum infection in vitro were studied in this paper, using an oocyst excystation assay, a cell culture model, and a free radical inhibition technique. H2O2 treatment at 500 and 1 000 μmol/L significantly inhibited excystation of bleach-treated oocysts (P＜0.01). Concentrations of H2O2 at 500 and 750 μmol/L resulted in a significant decrease in C. Parvum infection at 35.77% and 58.16% respectively, when compared with the untreated control at 48 hours postinoculation. Surprisingly, C. parvum infection were significantly increased by 22.21% to 39.33% following treatment with 50 (P＜0.01), 100 (P＜0.01) or 200 (P＜0.05) μmol/L H2O2, respectively. Stimulatory effect with treatment of 100 μmol/L H2O2 was most obvious, compared with the untreated control at 48 hours postinoculation. Effects of H2O2 on C. parvum at 24, 48 and 96 hours postinoculation were similar, the highest infection being infection at 48 hours postinoculation, with the maximum inhibitory effect being seen at 96 hours postinoculation. The stimulatory and inhibitory effects of H2O2 treatment on C. parvum infection were, to a certain extent, abolished in the presence of free radical scavengers, reduced glutathione or mannitol. These observations indicate that reactive oxygen species (ROS), such as H2O2, may play a role in the course of C. parvum infection. This is the first time to demonstrate a significant action of ROS in C. parvum infection in vitro.
作者： 刊期： 2001年第08期
Chickens were infected with CAV at one-day-old and 8 days later, the infected and uninfected chickens were vaccinated with La Sota vaccine. At 7, 14, 28 days post vaccination, the number of T cells and IgG, IgM and IgA antibody producing cells in Harderian gland and cecal tonsil, the content of IgG, IgM and IgA in tear, trachea fluid, intestinal fluid and bile as well as the hemoagglutination inhibition (HI) titer in tear and bile were detected. The results showed that the number of T cells and IgG, IgM and IgA antibody producing cells in Harderian gland and cecal consil, the content of IgG, IgM and IgA in tear, trachea fluid, intestinal fluid and bile as well as the HI titer in tear and bile post ND vaccination of CAV infected chickens were decreased significantly than those of uninfected vaccinated chickens. These indicated that the immune response function was markedly weakened in local mucosa of digestive and respiratory tract post ND vaccination of CAV-infected chickens.
作者： 刊期： 2001年第08期
AIM: To study dynamically the effect on the rabbits after fine carbon fiber powder was transplanted into their cranium. METHODS: After fine carbon fiber powder was injected into subdural cavity of rabbits, examination of histopathology and EEG were carried out. RESULTS: General observation: 1-104 weeks after injecting, no neuropathological changes concerning with the injecting of the carbon fiber powder were found in the rabbits of experimental group; Under optical microscopes: 1-2 weeks after injecting, slight inflammatory reaction in meninx was found and disappeared generally 4 weeks after injecting, no obvious fiber membrane was found around the carbon fiber, no significant differences were showed between the experimental group (EG) and the control group (CG); Examination of EEG: 1-2 weeks after injecting, slight abnormal changes of EEG in both two groups were showed, but no significant differences was found between them. 4 weeks after injecting, the EEG of the two groups was restored to their normal state before injecting. 1-8 weeks after injecting, no obvious epilepsy waves were showed. CONCLUSION: The fine carbon fiber powder showed excellent histocompatibility after injected into the subdural cavity of rabbits.
作者： 刊期： 2001年第08期
To clarify the mechanism in which signals to regulate food intake are transmitted from the gastro-intestinal system to the brain, we analyzed changes in hypothalamic neuronal histamine using cross-intestine parabiotic rats. Pairs of weight matched Lewis rats were sewn together in such a way as to form a common abdominal cavity. The small intestines of rats were transected and reconnected so that food eaten by one rat passed through a segment of its partner's intestine before returning to the intestine of the first rat. Concentrations of neuronal histamine were measured in microdissected hypothalami using radioimmnoassay. Sustained alteration of food intake were observed in both rats, one rat eating an average of 2.2 times (SE 0.15) as much as the other, without development of any significant difference in body weight after seven weeks. We found significant increase in hypothalamic neuronal histamine concentrations in the arcuate and tublomamelary nuclei of the hypophagic rats.These results are supportive of the theory that histamine acts in response to signals from the gut to regulate food intake.
作者： 刊期： 2001年第08期
Abstract］ AIM and METHODS:The recent work from our laboratory showed that ventral septal area (VSA) played a negative-regulatory central role in thermoregulation during endogenous pyrogen-induced fever. In order to further investigate the role of VSA in the antipyretic mechanism, we observed the effects of electrical stimulation of VSA on firing characteristics of thermosensitive neurons in preoptic anterior hypothalamus (POAH) by using extracellular microelectrode technique on 32 New Zealand white rabbits treated with interleukin-1 β (IL-1β) intracerebroventriculary (ICV). RESULTS:(1)Injection of IL-1β decreased discharging rate of warm-sensitive neurons in POAH. Electrical stimulation of VSA remarkably decreased thermosensitive coefficient of warm-sensitive neurons. (2)Injection of IL-1β caused increase in discharging rate of cold-sensitive neurons in POAH. Electrical stimulation of VSA remarkably increased thermosensitive coefficient of cold-sensitive neurons. CONCLUSTION:VSA may have an antipyretic effect through affecting the firing characteristics of thermosensitive neurons in POAH during IL-1β-induced fever.
作者： 刊期： 2001年第08期
In order to investigate the possible role of ONOO- in regulatory disorder of pulmonary arterial tension in endotoxic shock, the responses of rabbit pulmonary arterial rings (PARs) preincubated with ONOO- to endothelial dependent and receptor dependent relaxants acetylcholine (ACh) and adenosine diphosphate (ADP), endothelial dependent and receptor independent relaxant A23187, endothelial independent relaxant sodium nitroprusside (SNP) and α1-adrenoceptor agonist phenylephrine (PE) were observed in vitro in accumulative manner. Results were as follow: (1) Relaxations of PARs to ACh, A23187 and ADP were markedly impaired with shift of accumulative dose response curve of each agonist to the right. Inhibition of endothelial dependent and receptor dependent or independent relaxation by ONOO- was dose dependent. (2) ONOO- incubation inhibited SNP-induced relaxation in a dose dependent manner. Accumulative dose response curve of SNP was right shift to some degree depending on the doses of ONOO-. (3) Contractile response of PARs to PE varied with the different doses of ONOO-. In PARs preincubated with 0.5 mmol/L ONOO-, contractile reponse was significantly enhanced with shift of PE accumulative dose response curve to the left, while in PARs preincubated with 1.0 mmol/L or 2.0 mmol/L ONOO-, it was markedly reduced with right shift of PE accumulative dose response curve. (4) Vehicle of ONOO- had no effect on responses to every agonist, whereas decomposed ONOO- had minimal effect on the response to PE and ADP. In contrast, relaxation of PARs to ACh, A23187 and SNP were enhanced. These results suggested that direct effect of ONOO- on pulmonary artery may be a key factor contributing to regulatory disorder of pulmonary arterial tension induced by LPS and pulmonary hypertension in the early stage of endotoxic shock.
作者： 刊期： 2001年第08期
AIM: Music relieves anxiety and psychotic tension. This effect of music is applied to surgical operation in the hospital and dental office. It is still unclear whether this music effect is only limited to the psychological aspect but not to the physical aspect or whether its music effect is influenced by the mood or emotion of audience. To elucidate these issues, we evaluated the music effect on pain threshold by current perception threshold (CPT) and profile of mood states (POMC) test. METHODS: Healthy 30 subjects (12 men, 18 women, 25-49 years old, mean age 34.9) were tested. (1)After POMC test, all subjects were evaluated pain threshold with CPT by Neurometer (Radionics, USA) under 6 conditions, silence, listening to the slow tempo classic music, nursery music, hard rock music, classic paino music and relaxation music with 30 seconds interval. (2)After Stroop color word test as the stresser, pain threshold was evaluated with CPT under 2 conditions, silence and listening to the slow tempo classic music. RESULTS: Under litening to the music, CPT sores increased, especially 2 000 Hz level related with compression, warm and pain sensation. Type of music, preference of music and stress also affected CPT score. CONCLUSION: The present study demonstrated that the concentration on the music raise the pain threshold and that stress and mood influence the music effect on pain threshold.
作者： 刊期： 2001年第08期
AIM: To study the changes in capillarity of skeletal muscle during acclimation to high altitude, and explore the effects of a certain extent physical activity under hypoxia on capillary formation and the role of vascular endothelial growth factor (VEGF) in this process. METHODS: 48 Wistar rats were divided into 3 groups: Ⅰ normoxic control; Ⅱ hypoxia and Ⅲ hypoxia+exercise. Rats of Ⅱ and Ⅲ groups were subjected to hypobaric hypoxia for 5 weeks (23 h/d). They were first brought to simulated 4 000 m altitude, where rats of the Ⅲgroup were forced to swim for 1 h/d (6 d/week). Then the animals were ascent to 5 000 m. Biomicrosphere method was used to determine blood flow of skeletal muscle. The mean fiber cross-sectional area (FCSA), capillary density (CD) and capillary/fiber ratio (C/F) of red portion of the lateral head of the gastrocneminus were assayed by myofibrillar ATPase histochemistry. VEGF and its receptor KDR were assayed with immunohistochemistry method.RESULTS: By comparison with the normoxic control, 5-week hypoxic exposure resulted in a decrease in cross-sectional area of skeletal muscle fiber and an increase in CD, but the C/F remained unchanged. The blood supply to the gastrocnemius was not changed. After 5-week-exercise at high altitude, the muscle fibers did not undergo atrophy. CD, C/F, and the blood flow at rest increased significantly. VEGF protein was found primarily in the matrix between muscle fibers; KDR were shown mainly in endothelial cells of capillary. VEGF was more strongly stained in the skeletal muscle of hypoxia-exercise rats.CONCLUSION: Hypoxia itself can not induce neovascularization. While exercise during hypoxic exposure can lead to capillary formation. VEGF and KDR may play roles in it. New capillary formation benefits the blood supply, oxygen delivery and working performance at high altitude.
作者： 刊期： 2001年第08期
AIM:To investigate effect of recombinant human thrombopoietin on exsanguine thrombocytopenia mice. METHODS:Normal peripheral platelet counts were performed on sample obtained from the tail vein of purebred Babl/c mice including experimental and control groups before experimentation. rhTPO was injected into the mice by intraperitoneal injection once a day for 7 days. On the seventh and the fourteenth day, the mice were phlebotomized from the supra-obitalis vein in order to make exsanguine thrombocytopenia animal model. At the same time, we observed the biological activity of recombinant human thrombopoietin in vivo and the mice's death rate. RESULTS: On the seventh day and the fourteenth day, platelet counts of mice treated by rhTPO were higher than those by PBS (P＜0.05). Moreover the platelet counts of mice in experimental group of rhTPO showed increasing tendency following experimental days. In addition, death happened in two groups after those mice were phlebotomized from the supra-obitalis vein, but the death rate in negative control group was evidently higher than that in experimental group (P＜0.05). CONCLUSION:rhTPO had obvious biological activity in increasing platelet production, which resulted in the drop in thrombocytopenia mice's death rate.
作者： 刊期： 2001年第08期
Troglitazone, a member of the insulin-sensitizing thiazolidinediones, was shown to suppress voltage-dependent L-type calcium currents (ICa, L) in vascular smooth muscle cells. The present study was aimed to investigate whether troglitazone may inhibit ICa, L in rat cardiac ventricular myocytes and whether the extent of troglitazone-induced inhibition of the current may be different between normal and streptozotocin (STZ)-induced diabetic rats. Whole-cell patch-clamp techniques were applied in single rat ventricular myocytes. Troglitazone reduced the amplitude of ICa, L in a concentration-dependent manner in both normal and diabetic myocytes. However, the percent inhibition of ICa, L by troglitazone in concentration of 10-5mol/L measured at a holding potential of -50 mV was significantly greater in myocytes isolated from STZ-induced diabetic rats than in control rats (87%±8% vs 47%±5%, P＜0.005), without difference in voltage-dependency of the inhibition. These findings demonstrate that troglitazone-induced inhibition of ICa, L is augmented in STZ-induced diabetic cardiac ventricular myocytes, thus possibly playing a role in preventing intracellular calcium overload.
作者： 刊期： 2001年第08期
AIM: The aim of this study was to explore the expression of nerve growth factor(NGF) in spinal dorsal horn following crushed spinal cord injury. METHODS: The adult Srague-Dawley rat model of crushed spinal cord injury was established by the method in our laboratory, and intact spinal cord was used as control. The rats were sacrificed respectively after 24 hours, 7 days, and 21 days of operation, and the L3 spinal segments were removed out and fixed in 4% polyformaldehyde. The segments were sectioned into sections of 20 μm in thickness. The sections were stained with anti-NGF antibody by ABC method of immunohistochemistry technique. The immunoreactive intensity of NGF and the number of positive neurons as well as glial cells in dorsal horn were observed and counted under light microscope. RESULTS: The number of positive cells and immunoreactive intensity of NGF increased gradually in the dorsal horn at 24 hours, 7 days and 21 days following crushed spinal cord injury compared with control group (P＜0.01). CONCLUSION: These results indicated that NGF plays an important role in the postoperative reaction during the early period of the crushed spinal cord injury.
作者： 刊期： 2001年第08期
目的:旨在调查人β-防御素hBD基因在女性生殖道各段组织及妊娠相关组织的表达,探讨内源性抗菌肽在女性生殖道粘膜天然抵抗致病微生物侵袭机制中的作用.方法:采用逆转录多聚酶链反应法(RT-PCR)检测了正常女性生殖道各段组织及怀孕妇女生殖道相关组织中hBD-1、hBD-2的表达情况,以β-actin为阳性对照.结果:结果显示正常女性阴道、子宫颈、子宫内膜、输卵管、卵巢等5个组织中均发现有hBD-1 mRNA的表达,除阴道外其余组织也有hBD-2 mRNA的表达.绒毛膜、羊膜、绒毛、胎盘、脐带等5个妊娠相关组织中,hBD-1 mRNA在除羊膜外的各组织中均有表达,而hBD-2 mRNA则在胎盘、绒毛膜及绒毛组织中表达.结论:hBD在正常妇女生殖道及妊娠相关组织中广泛表达,提示其在维持正常妇女及孕妇生殖道内环境稳定、宿主天然抗感染机制中可能起着重要的作用.
作者：冯艳；潘小玲；黄宁；王伯瑶；冯云 刊期： 2001年第08期
目的:复制中风病痰热腑实证动物模型.方法: 用胶原酶脑内注射配合大鼠自体粪便灌胃的方法,复制中风病痰热腑实证中医证候大鼠模型.Wistar大鼠55只,雌雄各半,体重250-300 g,随机分4组:痰热腑实证模型组,以药测证组,单纯脑出血模型对照组,假手术对照组.参照文献向尾壳核缓慢注射胶原酶Ⅶ-肝素混合液1.25 μL.在复制脑出血模型前2 d起痰热腑实证组和以药测证组用10%自体粪便1 mL/100 g灌胃,每日1次,连续3 d,复制成痰热腑实证模型.以药测证组在灌胃自体粪便后第3 d开始,用通腑醒神胶囊灌胃,1 g/kg体重,每日1次,连续3 d;其余3组用等体积凉开水灌胃,每日1次,连续3 d.结果: 该模型大鼠有较明显的神经缺损体征和粪便干结、烦躁、鼻分泌物多、喉中痰鸣等热腑实证表现,脑含水量测定示该模型大鼠脑组织含水量明显高于单纯脑出血模型组(P＜0.01).通过以通腑化痰治法组方的通腑醒神胶囊治疗该模型大鼠,结果神经体征和粪便干结、烦躁、鼻分泌物多、喉中痰鸣等痰热腑实证表现明显减轻,脑组织含水量明显低于痰热腑实证模型组和单纯脑出血模型组(P＜0.01),说明模型是成功的.结论: 用胶原酶脑内注射配合大鼠自体粪便灌胃的方法可成功复制中风病痰热腑实证大鼠模型.
作者：孙景波；华荣；区永全；欧润妹；邓时贵 刊期： 2001年第08期
目的:探讨甘氨酸对内毒素所致心肌损伤的保护作用.方法:采用Langendord装置,应用Krebs-Henseleit(KH)液行主动脉逆灌注,动态观察离体心脏单相动作电位(MAP)和心肌张力曲线,并在实验的0、20、50、80 min收集灌流液,测定其中超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)、磷酸肌酸激酶(CK)、一氧化氮(NO)等的含量.结果:甘氨酸处理组对内毒素诱导心脏单相动作电位的D20、D50和心率等的变化有一定改善作用,但对心肌张力曲线的改善不明显,对内毒素诱导的SOD活性降低和MDA含量升高有抑制作用,可以减缓和降低心肌损伤引起的LDH和CK的释出,并且降低NO的释放等.结论:甘氨酸可以减轻内毒素所致心肌损伤、改善心脏功能状态,其作用机制可能与甘氨酸拮抗内毒素的作用及甘氨酸对血管内皮细胞、心肌细胞的保护作用有关.
作者：戚仁斌；陆大祥；王华东；王海华；颜亮；王彦平；付咏梅；李楚杰 刊期： 2001年第08期
目的:为开展防御素基因转基因以增强粘膜抗感染能力的实验研究,本实验构建真核重组表达载体pBK-CMV-rBD2以进行COS-7细胞转染表达实验.方法:用RT-PCR法(引物含有EcoRI、HindⅢ酶切位点)从肺组织总RNA中扩增出rBD2基因全长cDNA片段,并克隆于pBK-CMV.通过脂质体转染法将pBK-CMV-rBD2导入COS-7细胞,采用RT-PCR法和琼脂糖弥散法分别从mRNA水平和蛋白质水平检测rBD2的表达及活性.结果:所构建的真核表达重组载体pBK-CMV-rBD2转染COS-7细胞后呈现有效表达并具有活性.结论:提示通过基因转染的方法把防御素基因导入宿主细胞可能增强宿主粘膜抗感染能力,进一步的整体动物转基因抗感染实验研究具有可行性.
作者：周惠；黄宁；王伯瑶 刊期： 2001年第08期
目的:给动物口饲有机磷农药杀虫剂毒死蜱(CHP)先出现快速的降温相,然后出现发热相.众所周知精氨酸加压素(AVP)是体内一种重要内源性抗热物质.为此本实验观察了精氨酸加压素V1受体阻断剂(AVP V1阻断剂)对CHP引起的降温效应和发热效应的影响,旨在探讨CHP对体温影响是否与AVP有关.方法:实验用SD大鼠分为4组,每组7只,即对照组、CHP组、AVP V1阻断剂组和CHP-AVP V1阻断剂组.用体温遥测系统测量大鼠的体温和活动的变化.给药途径和药物剂量分别给大鼠口饲CHP 30 mg/kg(30 g/L玉米油)或玉米油1 mL/kg后,立即腹腔注射AVP V1阻断剂20 μg/kg(30 mg/L生理盐水)或生理盐水1 mL/kg.结果:给大鼠口饲CHP后可以引起快速的降温效应,给药后5 h后体温由给药前的(37.75±0.11)℃降低到(36.34±0.22)℃,平均降低达1.44℃;而给CHP后立即给AVP V1阻断剂时,大鼠体温由给药前的(37.55±0.17)℃降低到(37.19±0.14)℃,平均只降低了0.46℃.对照组动物在给药后,则出现短暂的应激性体温升高和活动增加现象.AVP V1阻断剂对CHP的发热相无明显影响.在实验中发现AVP V1阻断剂也可提高正常大鼠的体温,持续时间为2 h.结论:AVP V1阻断剂可以明显阻断CHP的降温效应,但对CHP的发热相无明显影响,提示AVP在CHP引起的降温过程中有重要作用.同时也证明内源性AVP在正常体温中可能有负调节作用.
作者：杨永录 刊期： 2001年第08期
目的:本实验以透明质酸(HA)、层粘连蛋白(LN)为指标,观察CCl4诱导大鼠中毒性肝损伤和肝纤维化后肝组织内储脂细胞变化和胶原的沉积及红花黄色素对其变化的影响,以探讨红花黄色素对中毒性肝纤维化防治作用的机制.方法: Wistar大鼠随机分为3组:①病理模型组:接受40% CCl4一橄榄油皮下注射,剂量为1.2 mL/kg体重,每周2次,共12周,与治疗组等容量生理盐水灌胃;②红花黄色素治疗组接受40% CCl4一橄榄油注射,方法、剂量同病理模型组,同时红花黄色素550 mg/kg体重灌胃,每日1次,共12周;③正常对照组接受橄榄油皮下注射,方法、剂量同上.各组分别于实验开始后,每隔3周随机处死5只大鼠,剩余大鼠于12周末全部处死.断头采血,分离血清,取1 g新鲜肝组织匀浆,用放免法测定大鼠血清和肝匀浆中HA、LN含量.肝组织V-G染色,用病理图像分析系统进行胶原定量分析.用透射电镜观察肝组织内储脂细胞变化.结果: 红花黄色素治疗组大鼠血清和肝匀浆中HA、LN含量及肝组织内胶原含量和储脂细胞变化显著低于病理模型组.结论: 红花黄色素能抑制肝脏储脂细胞增殖转化及HA、LN的合成,减少胶原沉积,有防治CCl4诱发中毒性肝纤维化的作用.
作者：白娟；王哲 刊期： 2001年第08期
目的:通过在体实验初步探讨参麦注射液对大鼠急性心肌缺血再灌注损伤的防治作用及相关机制.方法:通过结扎左冠状动脉前降支10 min,松开丝线复灌15 min,来复制大鼠急性心肌缺血再灌注损伤的模型.结扎的同时经股静脉缓慢推注参麦注射液,观察参麦注射液对再灌注性心律失常的影响,并测定心肌组织匀浆中MDA的含量、SOD的活力、Na+,K+-ATP酶和Ca2+-ATP酶的活性.此外,还通过光镜和电镜来观察参麦注射液对再灌后心肌超微结构的影响.结果:(1)模型组大鼠再灌注性心律失常的发生率为88.9%,持续时间为(142.8±73.4) s,参麦注射液可以使再灌注性心律失常的发生率降至33.3%,持续时间缩短至(31.7±20.1) s,两组之间有明显差异(P＜0.05).(2)模型组心肌组织匀浆SOD的活力为(27.287±3.449)×103 NU/g protein,参麦注射液治疗后SOD活力升高至(30.791±1.676)×103 NU/g protein,二者之间有显著差异(P＜0.05);模型组MDA的含量为(0.319±0.0515) μmol/g protein,参麦注射液治疗组为(0.262±0.0472) μmol/g protein, MDA的含量明显降低(P＜0.05).(3)模型组Na+,K+-ATP酶的活性为(0.3420±0.0391) mmol Pi*g-1protein*h-1,参麦注射液治疗后其活性升高至(0.3950±0.0265) mmol Pi*g-1protein*h-1,二者之间有显著差异(P＜0.01);模型组Ca2+-ATP酶的活性为(0.3450±0.0438) mmol Pi*g-1protein*h-1,参麦注射液治疗组为(0.4270±0.0624) mmol Pi*g-1protein*h-1,其活性明显升高(P＜0.01).(4)正常组和假手术组光镜显示心肌细胞排列整齐,横纹清晰,结构正常,电镜显示肌原纤维清晰,排列整齐,线粒体无肿胀,嵴密集,排列规则.模型组光镜显示心肌细胞排列紊乱,横纹不清晰,胞浆嗜酸性变,有明显的空泡变性,间质严重水肿,毛细血管内有红细胞淤积,电镜显示肌原纤维模糊不清,线粒体肿胀明显,嵴稀疏,排列紊乱.经参麦注射液治疗后,光镜显示心肌细胞排列整齐,横纹清晰,形态接近正常,电镜显示肌原纤维清晰,排列基本整齐,线粒体肿胀减轻,嵴排列也较规则.结论:参麦注射液能降低再灌注性心律失常的发生率,提高心肌组织中SOD的活力,降低MDA的含量,提高Na+,K+-ATP酶和Ca2+-ATP酶的活性,对缺血再灌注损伤引起的心肌超微结构的损伤有明显的保护作用.参麦注射液防治心肌缺血再灌注损伤的机制与增强机体清除氧自由基的能力,减轻脂质过氧化反应,拮抗自由基的毒性作用,维持离子泵的正常转运,减轻钙超载,从而维持了心肌细胞膜和线粒体的正常功能有关.
作者：李丽；黄启福 刊期： 2001年第08期
目的:Fall-39是Cathelicidine家庭中唯一存在人体内的抗菌肽,具有广谱抗菌活性和细胞毒性,对天然免疫与获得性免疫均发挥作用,因此在抗生素肽开发领域日益受到重视,本研究观察卡介苗能否增强人肺腺上皮细胞表达Fall-39.方法:根据GenBank中人Fall-39的cDNA序列资料,设计特异性引物,应用RT-PCR的方法检测mRNA的表达.超声击粹、蔗糖密度梯度离心及SDS抽提等步聚提取卡介苗胞膜蛋白,Sephadex G-75柱层析粗分其不同分子量蛋白.结果:卡介苗的确可刺激人肺腺上皮细胞Fall-39 mRNA的增强表达,这种增强表达具刺激物剂量和时间依赖性.BCG 100 mg/L刺激8 h时,Fall-39 mRNA表达量达到高.BCG胞壁蛋白经Sephadex G-75柱层析,获得的组分4(分子量范围约2.1-2.9 kD)刺激SPC-A-1细胞后Fall-39 mRNA的表达增强了8.5倍.培养细胞上清抗菌试验显示被刺激细胞培养上清的抗菌活性亦明显增强.结论:结果提示除细菌LPS外,BCG胞壁蛋白亦是Fall-39基因的激活因子,可诱导其基因的上调表达.
作者：朱玲；冯云；黄宁；王伯瑶 刊期： 2001年第08期
目的:ESAT-6是结核分枝杆菌的一种分泌性蛋白质,具有免疫保护作用,可作为抗结核病新疫苗的候选分子.而卡介苗由于缺失ESAT-6基因,不能产生该抗原蛋白.本研究旨在构建ESAT-6基因重组卡介苗,使其表达ESAT-6抗原,从而增强其免疫原性和免疫保护性,探索将该重组卡介苗用作新型结核病疫苗的可行性.方法:分别以卡介苗(BCG)和结核分枝杆菌H37Rv株基因组DNA为模板,经PCR扩增得到BCGα-抗原(α-Ag)信号肽序列和结核杆菌ESAT-6基因,将ESAT-6基因与大肠杆菌-卡介苗穿梭质粒载体pMV261重组,得到重组质粒pME.再将BCGα-Ag信号肽序列克隆至pME中,得到重组质粒pSME.后通过电穿孔法将pSME导入卡介苗中,并用抗性筛选、PCR扩增及打点杂交进行鉴定.结果:电转化后的卡介苗能在含10 mg/L卡那霉素的培养基上生长.以重组卡介苗总DNA为模板的PCR扩增得到阳性扩增带,大小与α-Ag信号肽序列加上ESAT-6基因的长度一致.打点杂交中探针与重组卡介苗显示阳性信号.结论:基因重组卡介苗构建成功,并有望分泌性表达结核分枝杆菌的免疫保护性抗原蛋白ESAT-6.该重组卡介苗既保留了卡介苗中原有的抗原,又增添了新的保护性抗原,且这种抗原可被广泛识别,能强烈诱导宿主T淋巴细胞增殖和持久的免疫记忆,因此ESAT-6重组卡介苗可能具有更好的免疫原性和保护力.本研究为改造卡介苗、发展新型结核病疫苗奠定了基础.
作者：陈玮；鲍朗；谢勇恩 刊期： 2001年第08期
目的:探讨阴阳虚证与糖皮质激素受体(GR)之间的关系.方法: 分别以临床典型阴虚阳虚患者、动物模型(大鼠)以及细胞模型(HL60)为研究对象,观察阴阳虚证时糖皮质激素受体(GR)的改变,并探讨中药上调GR的部分作用机制.结果: ①GR水平降低是阴阳虚证发展到一定阶段的共同病理基础之一;②阴阳虚证发展到一定阶段时,体内GR下降无器官特异性;③上调GR是参附汤、生脉饮被用于临床急救的重要作用机理之一;④参附汤、生脉饮上调GR的重要作用途径之一是增加细胞内GR mRNA的表达.结论: ①GR数量和/或活性的下降及其下降的幅度有可能作为中医虚证及其严重程度的重要微观指标之一;②有可能从生脉饮或参附汤中筛选出一种有效上调GR的成分(单体或混合物),并用于临床以虚证为主要表现的一类疾病(如肿瘤、风湿病、爱滋病等.)
作者：凌昌全；李敏；朱德增；陈吉吉；俞超芹；黄雪强；潭金兴；卢军华；徐仁宝；赵伟康 刊期： 2001年第08期
目的和方法: 以盐酸可乐定片(Cld)为对照,采用双盲双模拟临床试验,初步观察补脾益肺方(BFF,由党参、黄芪、白芍、当归等组成)对100例阿片类戒断综合征的疗效、不良反应和个体化用药方案.结果:①BFF对阿片类躯体和精神戒断症状均有疗效:BFF明显改善副交感兴奋类症状,如流泪、流涕、鼻充血、立毛肌收缩、腹泄、滑精和体重减轻等;BFF基本消除了感觉神经兴奋类症状,如腹痛、关节痛、骨痛、周身疼痛、蚁行感及寒热交错等;BFF一定程度上控制了精神紊乱方面的症状,如失眠、焦虑、烦燥、恐惧、抑郁等及由之导致的自伤自残或毁物伤人等极度表现.这些均达到或超过Cld的疗效.②抗戒断症状的时效关系:于治疗第0 d到10 d,从卧位血压、脉搏、体重、戒断症状量表、焦虑症状量表、副作用观察表、实验室检查、合并用药等的观察分析显示:BFF全程(第1 d到10 d)控制和延缓了阿片类躯体戒断症状(症状减轻和高峰后移);BFF全程改善了阿片类精神戒断症状,并达到或超过Cld的疗效.③调整戒断综合征的基础代谢状态:BFF在控制体重减低、甘油三酯降低、高密度脂蛋白升高、载脂蛋白A升高、载脂蛋白B降低及血糖降低等反映代谢紊乱的指标,均达到Cld的疗效.④副作用轻:Cld组低血压、窦性心动过缓、头晕、口干、耳鸣、晕倒、心慌、眩晕及便秘等不良反应在第2到6 d出现频率较高.而BFF组仅部分病例于第2到4 d出现呕吐、腹泻症状加重(与BFF组比较无差别),无其它不良反应.⑤抗戒断综合征药物的作用机理:BFF的处方组成和制备工艺是针对阿片类戒断综合征之辨证施治的科研思路摸索出来的.现代中药药理提示其主要中药具有针对症灶局部(非中枢性)的镇痛作用、针对病机(多环节)的镇静抗焦虑作用,能调节消化功能紊乱,能稳定循环功能波动,可增强机体防御能力等.仔细分析BFF的临床疗效均与这些药理效应一致.⑥个体化合并用药方案和注意事项:BFF疗效缓和稳定,无需个体化用药.但对合并症应充分治疗,如止吐、止泻、抗感染、维持水电解质平衡等,否则明显影响BFF的疗效.结论:与Cld比较,BFF治疗阿片类戒断症的疗效较好(全面、缓和、稳定、持久),不良反应较轻(副作用少、可预性强、程度轻微、维持短暂).
作者：蔡大勇 刊期： 2001年第08期
从基因治疗领域发展起来的核酸疫苗,能将编码抗原蛋白的外源基因(DNA或RNA)直接导入动物体细胞内,通过宿主表达系并诱导对该抗原的免疫应答.相对于传统疫苗,核酸疫苗具有许多优越性.它类似于天然感染过程,诱导机体产生全面的免疫应答,尤其是能诱导特异的CTL活性,有利于清除病毒等胞内感染、变性的肿瘤细胞等,用作治疗性疫苗其优越性更显突出.核酸疫苗具有便于改进,易于制备,价格低廉,保存方便等优点.
作者：游自立 刊期： 2001年第08期
目的:为增强问号赖型钩端螺旋体(017株钩体)内鞭毛抗原(flaB)与外膜抗原(ompL1)的免疫保护作用,克服保护性免疫持续时间短的难题,应用DNA重组技术以pcDNA 3.1为载体构建-表达flaB与ompL1基因的融合DNA疫苗载体,以期注入肌体后能延长保护期,激发机体长期免疫.在pcDNA3.1载体上,ompL1与flaB前后相连,前者较后者先获得递呈,与钩体天然递呈过程相似,并且双嵌合基因可诱导机体产生针对2个不同抗原表位的抗体,从而增加免疫应答能力.pcDNA3.1载体能表达融合蛋白,并且具有CpG motifs,因而可发挥佐剂作用.方法:提取017株钩体基因组DNA,参照钩体特有的高度保守的flaB与ompL1序列,设计二对四条引物:P1、P2、P3、P4.flaB与ompL1已被证实是两个良好的钩体疫苗候选抗原.通过聚合酶链反应(PCR),P1、P2引物扩增017株钩体的ompL1抗原基因;P3、P4引物扩增钩体的flaB抗原基因,分别经双酶切,以pcDNA3.1载体,将ompL1与flaB顺次同时定向嵌入同一pcDNA3.1中,获得重组质粒(ompL1与flaB),并将其转入JM109宿主菌中.在设计引物时,分别在P2、P3引物中,采用多个结构简单又不易形成折叠的甘氨酸作为接头,以维持其空间构象,不影响自然折叠,保持天然活性.结果:经酶切鉴定证实:有一1.8 kb片段插入载体,进一步经酶切证实这一1.8 kb可被酶切为9.6 kb及8.5 kb的片段,与预期的片段大小一致.以嵌合重组质粒DNA为模板,经PCR证实:P1、P2引物扩增出9.6 kb片段;P3,P4引物扩增出一8.5 kb片段.重组质粒经DNA序列检测,其中ompL1与flaB序列与文献报道的完全一致.结论:表达017钩体flaB与ompL1抗原的融合蛋白的DNA疫苗载体构建成功,从而为下一步表达复合功能蛋白,进行钩体免疫保护性研究打下了基础.
作者：王敏；戴保民 刊期： 2001年第08期
目的:现已确定动脉粥样硬化是一种炎症疾病,单核/巨噬细胞在动脉内膜下集聚是导致动脉粥样硬化损伤发展的基础;血流低切应力促进其损伤的发展.趋化细胞因子MCP-1对单核/巨噬细胞起强力趋化和激活作用,并有报告切应力可诱导其基因的表达.新近报道在层流作用下IL-8亦是单核/巨噬细胞与内皮细胞相互作用的重要调节因子.本研究旨在观察切应力诱导人脐静脉血管内皮细胞IL-8 mRNA的表达,探讨Toll/NF-κB信号转导通路对其基因激活的介导作用.方法:4.2 dyn/cm2层流切应力处理人脐静脉血管内皮细胞,提取总RNA,应用RT-PCR和Northern杂交技术检测切应力作用不同时间后IL-8、TLR-2和TLR-4 mRNA的表达.免疫印迹技术检测IκB磷酸化和降解.免疫荧光细胞化学染色检测NF-κB胞核易位.结果:切应力作用0.5 h后IL-8 mRNA表达逐渐增高,2 h时达到很高水平.胞浆蛋白IκB和磷酸化IκB的免疫印迹显示4.2 dyn/cm2层流切应力作用10 min后磷酸化IκB水平即显著增加,30 min后又逐渐下降,与之相应IκB含量随切应力作用时间而逐渐下降,到1 h时几乎测不出,表明在切应力作用下内皮细胞胞浆NF-κB抑制因子IκB发生了磷酸化和降解.NF-κBp65亚基免疫细胞化学染色显示在4.2 dyn/cm2切应力作用0.5 h后内皮细胞核逐渐着色,1.5 h时细胞核几乎全着色,表明胞浆NF-κB在切应力作用下发生了易位,从胞浆进入胞核.RT-PCR检测和Northern杂交分析显示内皮细胞固有表达TLR-2和TLR-4 mRNA,在4.2 dyn/cm2切应力作用1 h后TLR-4 mRNA的表达显著增强,而TLR-2 mRNA的表达变化不明显.结论:流体切应力可诱导血管内皮细胞表达IL-8,活化转录因子NF-κB.在切应力作用下内皮细胞TLR-4 mRNA表达增强,提示Toll/NF-κB信号转导通路可能参与动脉粥样硬化性炎症反应.
作者：梁峰；黄宁；王伯瑶；陈槐卿；吴立志 刊期： 2001年第08期
目的:许多证据表明:利用转基因植物作为生物反应器规模化生产高附加值的药用重组蛋白质,其技术可行、成本低廉、效益显著,具有广阔的应用开发前景.本研究拟运用植物转基因技术,尝试构建高效表达hBD-2的植物生物反应器的可能性.方法:将C端带Myc和6xHis双标记的hBD-2的重组DNA克隆到植物真核表达载体pCAMBLA1303中,构建C端带Myc和6xHis双标签的hBD-2的重组植物真核表达载体;并将此载体转化根癌农杆菌LBA4404,利用卡那霉素进行抗性基因筛选,确定根癌农杆菌的阳性克隆;通过根癌农杆菌将外源基因转入马铃薯植物愈伤组织细胞,利用潮霉素抗性基因筛选及PCR检测,监测hBD-2在其中的遗传表达.结果:(1)酶切分析、PCR验证和DNA测序等证据表明,C端6个组氨酸的hBD-2已被正确地插入pCAMBIA1304载体中CaMV 35S和Nos终止子之间,成功构建了重组hBD-2/His基因的植物表达载体rpCAMBIA1304/hBD-2/His.(2)rpCAMBIA1304/hBD-2/His已成功地转化根癌农杆菌LBA4404,阳性克隆菌株已获得.(3)潮霉素抗性进行筛选及PCR检测结果提示:hBD-2 cDNA基因已整合到马铃薯植物愈伤组织的基因组中.结论:结果表明,重组hBD-2/His基因的植物表达载体通过根癌农杆菌介导进入了马铃薯细胞植物愈伤组织.利用植物转基因技术,构建高效表达hBD-2的植物生物反应器具有可能性.
作者：蔡绍晖；余懋群；杨小娟；马欣荣；黄宁；王伯瑶 刊期： 2001年第08期
目的和方法:本文对大鼠进行第三脑室和脑腹中隔区插管,用数字体温计测量大鼠的结肠温度,用放射免疫分析法测定脑中隔区AVP含量,观察脑中隔区精氨酸加压素(arginine vasopressin, AVP)在大鼠促肾上腺皮质激素释放激素(CRH)性发热机制中的作用.结果:脑室注射CRH(2.5 μg, 5.0 μg)引起大鼠结肠温度显著升高,脑室注射5.0 μg CRH也能明显增高脑中隔区AVP的含量.脑腹中隔区注射AVP V1受体拮抗剂本身并不导致大鼠结肠温度明显的改变,但能显著增强脑室注射CRH引起的发热反应.而且腹中隔区注射AVP也能显著抑制大鼠CRH性发热.结论:发热时CRH是引起脑腹中隔区AVP释放的因素之一,脑腹中隔区内源性AVP抑制中枢注射CRH引起的体温升高.
作者：王华东；王彦平；胡巢凤；戚仁斌；陆大祥；颜亮；李楚杰 刊期： 2001年第08期
目的:通过构建两种SV40系列的重组载体,即N端带Flag标签的重组hBD-2基因和C端带Myc和6xHis双标签的重组hBD-2,并转染COS-7细胞,探索建立既能持续有效地表达和分泌hBD-2、其表达产物又易于被检测和分离纯化的哺乳类动物细胞表达系统的可能性及技术路线.方法与结果:(1)将hBD-2全长cDNA片段插入带有报告基因的真核表达质粒pCMV.tag2B构建出hBD-2的上游带Flag报告基因的重组表达载体pCMV.tag2B/hBD-2,经DNA测序证明:hBD-2cDNA片段的插入方向和其全长cDNA的碱基组成顺序均准确无误.(2)hBD-2全长cDNA片段插入编码双重标签基因Myc和6xHis的真核表达质粒pcDNA3.1/Myc-His(+),构建出C端带Myc和6xHis双重标签的重组真核表达载体pcDNA3.1/Myc-His(+)/hBD-2.DNA测序结果表明:hBD-2cDNA片段插入方向和全长cDNA的碱基组成顺序均准确无误.(3)采用脂质体转染法分别将以上两种重组真核表达载体导入COS-7细胞,分别从mRNA和蛋白质水平分析hBD-2的表达情况.并检测经重组真核表达载体pcDNA3.1/Myc-His(+)/hBD-2转染细胞的可溶性蛋白及其培养上清的抗菌活性.(4)采用RT-PCR法对经带Flag报告基因的重组表达载体pCMV.tag2B/hBD-2转染的COS-7细胞的总RNA进行RT-PCR扩增,结果扩增出一条约240 bp的cDNA片段,其大小与预测相符.经Western blot法检测到细胞可溶性蛋白在约10 kD处有强反应条带显示.(5)用特异性引物(hBD2p6/hBD2p7)对经pcDNA3.1/Myc-His(+)/hBD-2转染的COS7细胞的总RNA进行RT-PCR扩增,用抗标签基因His表达产物的特异性抗体,经Western blot法检测到细胞蛋白在约10 kD处有强反应条带显示,其大小与该重组质粒结构中由pCMV启动子驱动的基因片段有可能表达成肽链的分子量(10.1)相符.双层肉汤琼脂平板扩散法抑菌试验结果表明:分别与正常COS-7细胞的细胞可溶性蛋白及其培养上清比较,转染pcDNA3.1/Myc-His(+)/hBD-2的COS-7细胞的细胞可溶性蛋白及其培养上清在金黄色葡萄球菌质控菌株25 923的平板上形成明显的抑菌环.结论:(1)N端带Flag标签基因重组真核表达载体pCMV.tag2B/hBD-2已被成功构建和转染COS-7细胞.通过RT-PCR法和Western blot法证实hBD-2基因已在COS-7细胞内持续有效地表达.(2)C端带双重标签基因Myc和6xHis的真核表达质粒pcDNA3.1/Myc-His(+)/hBD-2已被成功构建和转染COS-7细胞.通过RT-PCR法、Western blot法和体外抗菌实验证实,该系统不仅能有效地表达和分泌具抗菌活性hBD-2,也为其表达产物的检测、分离纯化提供了必要条件.
作者：蔡绍晖；黄宁；王伯瑶 刊期： 2001年第08期
目的:BEVS(Baculovirus expression vector system)杆状病毒表达系统是近年涌现的适用范围广、表达效率高的真核表达系统之一.本研究旨在探索应用该系统高效表达hBD-2的可行性及技术路线.方法:(1)利用引物hBD2p6和hBD2p9经PCR扩增,从原质粒的pcDNA3.1/Myc-His(+)/hBD-2中扩增出带有翻译起始密码ATG和终止密码TGA的完整重组hBD-2/His基因片段,通过双酶切,使该片段两端分别带有ScaI和KpnI两种限制性内切酶粘性末端;将此外源性片段插入转移质粒pAcGHLT-A的多克隆位点中的ScaI和KpnI位点上,使其处在多角蛋白基因强启动子的控制之下.(2)经重组pAcGHLT-A/hBD-2/His转移载体和AcNPV DNA共转染Sf21细胞,并在细胞内进行同源重组,形成重组病毒rAcNPVhBD-2/His.于共染后第5 d,收集各组细胞的培养上清,用终点稀释分析法检测共转染效率.经过多轮感染上清反复感染Sf21细胞和终点稀释分析法验证,获得高滴度的重组病毒.(3)用特异性抗-His抗体,经Western blot检测细胞及上清hBD-2/His的表达情况.结果:经酶切鉴定和测序分析证实:C端带Myc和6个多聚组氨酸双重标志的重组hBD-2已正确地插入BEVS系统的转移载体中,构建成重组转移载体pAcGHLT-A/hBD-2/His.经过多轮感染上清反复感染Sf21细胞及终点稀释分析法验证,高滴度的重组病毒rAcNPVhBD-2/His已被获得.Western blot检测结果提示Sf21细胞已表达和分泌6xHis-GST-hBD-2-6xHis融合蛋白.结论:本研究结果支持利用BEVS杆状病毒-昆虫表达系统作为高效表达重组hBD-2生物反应器的设想.
作者：蔡绍晖；黄宁；王伯瑶 刊期： 2001年第08期
目的:研究热灌注液经肝动脉灌注对犬肝组织学及肝、肾功能的影响,探讨介入性热化疗临床应用的安全性.方法:经犬肝动脉灌注入60℃生理盐水180 mL,持续30 min.于热灌注前及热灌注后24、48、72、120、168、216 h抽取静脉血,进行活体肝穿刺,分别进行生化和病理细胞检查,比较热灌注前后肝脏组织及肝、肾功能的变化.结果:1.谷丙转氨酶、谷草转氨酶于灌注后24 h开始升高,72 h达高峰(P＜0.01),216 h后恢复正常;总胆红素、白蛋白、γ-谷氨酰基移换酶无明显变化(P＞0.05);2.肝组织可见一过性损伤,主要表现为淋巴细胞浸润,肝细胞明显浑浊、肿胀,但未发现肝细胞坏死,热疗后168 h后恢复正常;3.肾功能无明显变化(P＞0.05).结论:介入性热疗对犬肝组织及肝、肾功能有一过性影响,但未发现造成组织细胞坏死,并且这种副反应在短期内可以完全恢复.
作者：郭卫平；刘燕；张洪新；梁志会 刊期： 2001年第08期
目的:对麦冬提取物抗心肌缺血的药效学观察做到标准化、序列化和规范化,为临床提供有效、安全、稳定,使用方便的新药制剂.方法: 采用犬冠状动脉结扎模型,设立模型组、恬尔心组及麦冬提取物大、中、小3个剂量组,应用十二指肠给药的途径,通过心外膜心电图、心肌酶谱及形态学等指标来观察麦冬提取物对缺血心肌的保护作用.结果: (1)心外膜心电图标测结果:结扎30 min时,心外膜心电图标测Δ-ST均有明显变化,各组差别无显著(n=5,P＞0.05).麦冬提取物大剂量组(LD)及中剂量组(MD)用药后1 h Δ-ST分别由给缺血30 min时的45.40±16.42及38.50±10.85降至38.30±9.28和38.30±9.28.随着药物作用时间延长至3 h,麦冬提取物大剂量组及中剂量组Δ-ST进一步分别下降了28.20±9.57及24.00±3.88,Δ-ST相对变化率为1.61±0.17(n=5,P＜0.01)和1.60±0.38(n=5,P＜0.05).(2)心肌酶谱(CK、CK-MB)观察结果:结扎前心肌酶谱已有变化,但各组之间并无明显差别(n=5,P＞0.05),实验观察结束时,各组心肌酶谱均明显变化.CK-MB,麦冬提取物大剂量组和中剂量组心肌酶谱与模型组比较均显著差异(n=5,P＜0.01).CK,麦冬提取物大剂量组与模型组比较均显著差异(P＜0.01).而中剂量组心肌酶谱与模型组比较均明显差异(n=5,P＜0.05).酶谱(CK、CK-MB)的自身变化率比较显示:大剂量组与模型组相比,显著差异(n=5, P＜0.01),而中剂量组与生理盐水组比较,P＜0.05.(3)犬急性心肌梗死范围(N-BT染色)测量结果:麦冬提取物大、中两个剂量组心肌梗死区的绝对重量以及梗死区占心室的百分比与M模型组相比,显著差异(n=5,P＜0.05).(4)透射电镜超微观察显示:模型组表现出缺血心肌典型超微结构特征:如线粒体、细胞核和肌原纤维核浆的改变及增生活跃间质细胞所分泌的胶原.与模型组比较小剂量组心肌超微结构的改变虽有所减轻,但仍发生明显的线粒体及细胞核损伤.中剂量组心肌超微结构线粒体及细胞核虽有部分、一定程度的损伤,但与模型组相比,有一定的改善.大剂量组心肌超微结构的改变发生了明显的改观.结论: 麦冬提取物有明显地抗心肌缺血的作用,并呈现出一定的量效关系.
作者：程金波；卫洪昌；章忱；朱晓梅；吕嵘 刊期： 2001年第08期
目的:探讨钩端螺旋体病的流行因素与防治措施.方法:用公认的方法进行鼠情调查和对病人、动物进行病原学、血清学研究.结果:①疫情:在监测的15年中湖南全省年均发病率为9.86/10万,此期间有2个发病高峰年;长沙地区则年均发病率为19.06/10万,近全省发病率的2倍.1992年-2000年9年中有7年发病率高于全省发病率.②鼠情监测:共布鼠夹11 803只,捕鼠747只,鼠密度为6.33%,流行前期、后期鼠密度差异无显著.③病原学监测:对717例钩体病人采血作钩体培养,阳性124份,阳性率17.29%,至少分属8个血清群,以流感伤寒和秋季热群为主;从416只鼠的肾中分离出49株钩体菌株,阳性率为11.78%,至少分属5个血清群,以黑线鼠携带黄疸群为主,其次为黄毛鼠携带爪哇群,占14.29%;从40只家犬肾中分离钩体菌9株,带菌率为22.50%,鼠与犬带菌率差异无显著.对72头猪117只蛙取肾培养,未获阳性结果.④血清学监测检测钩体病人双份血清抗体,黄疸出血群、犬群、澳洲群、波摩那群、流感伤寒群、七日热群6个群抗体均数差异有非常显著;爪哇群其抗体差异显著;在对动物血清抗体调查中,检测了犬、猪、牛分别为40、30、25头.具有某一抗体阳性的分别为60%、53.33%和84%,其差异有显著.犬抗体分属6个血清型,以澳洲为主;猪则分属5个血清型,以巴达维亚为主;牛分属11个血清型,以巴达维亚为主.结论:①长沙地区在15年监测中平均发病率为全省的2倍左右,除个别年度外几乎每年发病率都较高,是湖南省的典型代表.②鼠类尤其是优势鼠种黑线姬鼠、犬、猪和牛是主要传染源.③病人血培养结果显示,感染菌群非常复杂,至少有8个钩体菌群.④用现行的四价菌苗接种易感人群其作用有限.
作者：郭绶衡；吴子贵；李俊华；李立新；李伟；朱国平 刊期： 2001年第08期
目的:通过不同型钩体DNA序列分析筛选钩体基因疫苗的候选株,并从基因水平解释钩体血清型的特异性.方法:(1)酚、氯仿抽提法提取017株与56610株钩体基因组DNA.(2)分别以017株与56610株钩体基因组DNA为模板,PCR法扩增内鞭毛蛋白(flaB)基因.(3)T/A快速克隆法将两个flaB基因的PCR产物连接与Teasy-vector载体.(4)α-互补、PCR法初步筛选重组质粒.(5)碱裂解法小量提取质粒、酚切分析进一步鉴定重组质粒,确定外源基因插入方向.(6)双脱氧终止法对重组质粒的克隆化flaB基因进行测序.(7)应用软件对所测序列进行酶切位分析及编码蛋白的预测.(8)与已发表的钩体内鞭毛基因进行比较.结果:经鉴定获得了两个重组质粒pDHTF017与pDHTF610.pDHTF017中flaB基因为反向插入,pDHTF610中的flaB基因为正向插入.两个重组外源flaB基因长度均为852 bp,编码蛋白含283个氨基酸,预测分子量为31.5 kD左右.两个基因的限制性内切酶位点相似,含多个常见酶切位点.几个血清型钩体的flaB基因比较发现型间同源性很高(90%-99%),变异通常发生在固定位点,各型选择不同的固定位点.结论:(1)重组质粒pDHTF017与pDHTF610中的外源基因均为钩体的一种flaB基因; (2)flaB基因保守性强,可作为基因疫苗的靶基因;(3)七日热型钩体56610株的flaB基因有望成为核心抗原基因,可进行钩体疫苗的研究开发.
作者：何泼；戴保民；乔中东；游自立；方之茂 刊期： 2001年第08期
目的:利用抑制消减杂交技术(suppression subtractive hybridization, SSH),比较致病与非致病钩体之间基因组差异,筛选致病钩体特有的基因片段,以分析钩体毒力相关基因.方法:以赖型钩体017株为tester,非致病性patoc Ⅰ株为driver进行SSH.用3种四碱基内切酶RsaⅠ、HaeⅢ、AluⅠ分别酶切基因组DNA,选择适内切酶消化产物(小于2 kb的片段),连接特殊设计的Adaptor进行消减杂交的PCR,得到消减混合物,与T/A克隆载体连接,转化JM109建立差异消减文库,经PCR和Southern blotting筛选鉴定阳性克隆,进而对部分片段进行测序和相似性分析.结果:AluⅠ适于将钩体酶切成用于SSH技术的小片段;经PCR和Southern blotting筛选鉴定得到25个致病钩体中特异存在的阳性克隆,阳性率达到62.5%;序列测定显示插入片段大小为149-1 506 bp,平均480 bp, GC含量从26.30%到51.98%不等,平均36.63%;相似性分析其中6个片段无任何相似性匹配,18个有一定相似性的序列可分为两类:与外膜蛋白或假想蛋白相关和与生化代谢修饰的酶相关.20个序列已被GeneBank收录,收录号分别为AF300873-AF300877,AF325807-AF325821.结论:SSH是一种有效、灵敏的、高特异的比较分析基因组差异的方法,对钩端螺旋体致病基因的筛选、基因组分化、分子遗传研究等具有重要意义;利用SSH技术筛选得到的致病钩体特有消减片段很可能与致病钩体的分子进化和毒力有关,也为我国特有微生物的基因资源开发和基因工程疫苗的设计提供给了重要信息.
作者：胡昌华；鲍朗；谢勇恩；李学敏；陈玮；张会东 刊期： 2001年第08期
目的和方法:细胞因子在伤口愈合过程中有重要的影响,本研究主要观察中药珠香散对阿霉素攻击后大鼠皮肤伤口愈合过程中多种细胞因子的影响.实验分为4组,(1)对照组;(2)模型组;(3)模型+珠香散组;(4)模型+bFGF组.除对照组外其它3组于实验前4 d尾静脉注射8 mg/kg体重阿霉素,在第5 d时背部做一直径为1.5 cm的全厚皮切口,分别给予生理盐水、珠香散和bFGF.观察各组大鼠伤口愈合时间、伤口抗断裂强度、创面组织学观察、伤口TNFα、PGF2水平和局部TGFβ1 mRNA的表达.结果:对照组平均愈合时间为(17.18±1.72) d、组织抗断裂强度(TBS)为(8.68±1.56)kg/cm2,阿霉素攻击后愈合时间延长为(19.80±3.19) d、TBS为(6.36±1.36)kg/cm2,与对照组相比分别P＜0.05;珠香散和bFGF治疗后愈合时间则分别为(17.36±2.29) d和(18.60±2.31) d,接近对照组;TBS分别为(8.22±1.82) kg/cm2,和(7.56±1.72) kg/cm2,有较大的提高,珠香散治疗后TBS明显高于模型组,P＜0.05.这些结果证实珠香散可促进伤口的愈合.局部TNFα水平表明创面第3 d时模型组TNFα水平明显低于对照组,分别为(9.95±2.16) ng/g和(4.62±1.86) ng/g,P＜0.05;在第7 d时,对照组TNFα升高为(11.09±3.87) ng/g,而模型组为(7.17±1.92) ng/g,明显低于对照组,P＜0.05;珠香散治疗后TNFα水平明显升高(11.23±3.23) ng/g,明显高于创面第3 d时(5.53±2.50) ng/g和同期模型组的水平,分别P＜0.05.各组创面的PGE2在第3 d、7 d时无明显差异.伤口TGFβ1mRNA表达结果显示珠香散治疗后3 d TGFβ1mRNA表达为(+ +),高于模型组(+),在创面第7 d成纤维细胞表达增强至(+ + +);bFGF组在伤口3 d TGFβ1表达也明显增加为(+ +);伤口7 d创面表达(+ + +),持续到伤口第15 d,肉芽组织仍有表达至(+ +).组织学观察显示模型组在伤口的第3 d时,创面内嗜中性粒细胞(PMN)数量少,而模型+珠香散组在伤口的第3 d创面有大量的嗜中性白细胞,创面细胞数量增多,主要为单核巨噬细胞和成纤维细胞,表皮细胞向伤口区延伸较多;模型+bFGF组在伤口3 d创面也有大量的PMN,创面成纤维细胞和单核巨噬细胞数量明显增多.结论:提示珠香散可能通过吸引白细胞,激活吞噬细胞释放细胞因子,并通过增加内源性细胞因子的表达,促进伤口细胞增殖和胶原合成,从而加快伤口的愈合.
作者：李萍；杨丽彩；盛巡；徐宝婴；黄启福 刊期： 2001年第08期
目的:观察解热醒神(JRXS)注射液对内毒素(endotoxin, ET)性发热1 h家兔体温、下丘脑(hypothalamus, HP)环一磷酸腺苷(cyclic adenosine monophosphate, cAMP)及白细胞介素-1β(interleukin-1β,IL-1β)含量的影响.方法:20只新西兰家兔随机分为4组.NS组:NS 1.5 mL/kg iv;JRXS组:JRXS 1.5 mL/kg iv; ET组:ET 0.5 μg/kg iv; JRXS+ET组:JRXS 1.5 mL/kg iv, 30 min后,ET iv(剂量同ET组),观察1 h后,穿刺小脑延髓池抽取脑脊液,然后取下丘脑组织,并分别按cAMP(上海中医药大学同位素室)及IL-1β放免试剂盒(中国人民解放军总医院科技开发中心产品)说明书进行含量测定.观察指标:平均体温变化曲线;大体温上升高度(△T℃);体温反应指数(thermal response index, TRI);下丘脑、脑脊液的cAMP含量;下丘脑的IL-1β含量.所有实验数据用均数±标准差(眘)trtJRXS(P0.05)JRXSETJRXS+ETT(0.10.13)TRI10.36.62cAMP(1.36.27) nmol/gcAMP(14.40.69) nmol/LIL-1(2.90.37) ng/gETT(0.40.11)TRI11.78.79cAMP(2.90.40) nmol/g cAMP(32.10.51) nmol/LIL-1(6.08.79) ng/g(P0.01)cAMPcAMPIL-1(r1=0.918P0.05r20.926P0.05r3=0.868P0.01)JRXScAMP,IL-1
作者：张丹卉；蒋玉凤；黄启福；贾旭；严京；路雪雅 刊期： 2001年第08期
目的:探讨大黄虫庶虫丸治疗实验性大鼠脑出血的可能机理.方法: 采用胶原酶与肝素复合注入大鼠尾状核,制成大鼠脑出血模型.将动物分成正常组、假手术组、模型组、大黄虫庶虫丸治疗组,分别观察其对脑出血大鼠脑组织病理形态学变化、神经功能缺损积分、脑组织含水量;脑组织生化指标一氧化氮(NO)、内皮素(ET)、总抗氧化能力(T-AOC)、兴奋性氨基酸(EAA)/抑制性氨基酸(IAA)改变;并进一步运用了RT-PCR的方法观察了其对脑出血大鼠脑组织IL-1 β mRNA、iNOSmRNA、凝血酶受体(TR)mRNA表达的影响. 结果: 大黄虫庶虫丸能改善大鼠脑出血模型的病理形态结构的损害;降低脑含水量;改善脑出血大鼠的神经功能缺损;降低脑出血大鼠脑组织的NO、ET的含量;改善脑出血大鼠脑组织的EAA/IAA的失衡;提高大鼠脑组织的总抗氧化能力;下调因出血而致的脑组织IL-1 β mRNA、iNOSmRNA、凝血酶受体(TR) mRNA过度表达.结论: 大黄虫庶虫丸可能与降低脑出血大鼠脑组织的NO、ET、EAA的升高而致的细胞毒性,提高大鼠脑组织的总抗氧化能力;降低脑组织(TR)mRNA过度表达,以减轻凝血酶对脑组织的毒性损害;降低脑组织IL-1 β mRNA过度表达,以减轻炎性介质对组织的损害有关.
作者：戴高宗；陈汝兴；卫洪昌；朱晓梅 刊期： 2001年第08期
目的:旨在分离和鉴定刺激人肺腺上皮细胞(SPC-A-1)β-防御素-1(hBD-1)基因表达的卡介苗胞壁活性蛋白,为开发增强粘膜抗菌肽基因表达的免疫增强剂以防治粘膜感染的研究打下基础.方法:采用超声破碎细胞、蔗糖密度梯度离心、Sephadex G-150柱层析等方法分离卡介苗胞壁蛋白质组份;应用逆转录聚合酶链反应(RT-PCR)法和Northern杂交检测SPC-A-1细胞hBD-1 mRNA的表达;采用琼脂糖弥散杀菌法检测SPC-A-1细胞培养上清的抗菌活性.结果:RT-PCR和Northern杂交分析均证实卡介苗刺激SPC-A-1细胞hBD-1 mRNA的表达增强了2倍.卡介苗胞壁蛋白Sephadex G-150层析所得6个组分中,组分5(分子量范围约21×104-29×104)诱导SPC-A-1细胞hBD-1 mRNA的表达增强4倍,其培养上清的抗菌活性显著增强.这种诱导作用具剂量和时间依赖关系.结论:结果提示卡介苗胞壁蛋白是β-防御素基因诱导表达的一新的激活因子.
作者：冯云；黄宁；吴琦；王伯徭 刊期： 2001年第08期
目的:观察清开灵(QKL)注射液对内生致热原(endogenous pyrogen, EP)性发热家兔下丘脑与脑脊液cAMP(cyclic adenosine monophosphate)及腹中隔区(ventral septal area, VSA)精氨酸加压素(arginine vasopressin, AVP)含量的影响,探讨中药对发热体温的正负调节机制.方法:动物选用健康雄性封闭群新西兰白兔.EP制备采用家兔全血加精致大肠杆菌内毒体外培育法.实验分两部分进行,第一部分观察QKL对EP性发热的解热效应.将20只家兔随机分为4组.NS组:NS 1 mL/kg iv;QKL组:QKL 1 mL/kg iv;EP组:EP 1 mL/kg iv; QKL+EP组:QKL 1 mL/kg iv, 30 min后,EP 1 mL/kg iv, 观察3 h.第二部分观察QKL对EP性发热1 h下丘脑cAMP及VSA中AVP含量的影响,动物数、分组及给药同第一部分.观察指标:平均体温变化曲线;大体温上升高度(△T℃);体温反应指数(thermal response index, TRI);下丘脑的cAMP含量;VSA的AVP含量.所有实验数据用均数±标准差(眘)trtQKL(P0.05)QKLEPEPT(1.10.16)TRI3(12.28.59)EP+QKLT(0.78.17)TRI3(8.41.84)(P0.01)QKLEP1 hcAMPEP+QKLcAMP(1.11.29) nmol/gcAMP(14.23.57) nmol/gEPcAMP(3.32.34) nmol/gcAMP(34.33.46) nmol/g(P0.01)QKLEP1 hAVPEP+QKLAVP(16.13.89) ng/gEPAVP(39.94.88) ng/g(P0.01)cAMPcAMPAVP(r1=0.852P0.05r2=0.861P0.05r3=0.840P0.05)
作者：张丹卉；蒋玉凤；黄启福；贾旭；严京；路雪雅 刊期： 2001年第08期
动脉粥样硬化症(AS)是严重危害人类生命的主要疾病之一.基于我室对含川芎的复方防治AS的前期研究和川芎兼治AS的临床报道,本实验研究了川芎防治AS的作用机制.
作者：梅家俊；蔡大勇；陈振发；叶根梅；田文红 刊期： 2001年第08期
目的:通过离体实验初探人参皂甙Rg1的肠内菌代谢产物Rh1对正常小鼠免疫细胞功能的影响,并比较二者的差异.方法:用MTT比色法测定脾脏T、B淋巴细胞增殖能力.用中性红比色法测定腹腔巨噬细胞的吞噬能力.用Griess法测定腹腔巨噬细胞释放NO的能力.Rh1与Rg1的终浓度分别为0.1 mg/L、1 mg/L、10 mg/L.结果:(1)Rh1在低、中、高浓度对脾脏淋巴细胞增殖均有直接的促进作用(P＜0.05或P＜0.01),各浓度Rg1仅有促进脾脏淋巴细胞增殖的趋势,无明显差异.二者相比,低、中、高浓度Rh1的作用均比同浓度Rg1强(P＜0.05或P＜0.01).(2)3种浓度的Rh1对ConA诱导的T淋巴细胞增殖呈浓度依赖性抑制,且有显著差异(P＜0.01或P＜0.05),Rg1只在高浓度有明显抑制作用(P＜0.05),低、中浓度Rg1虽有作用趋势,但无明显差异.二者相比,Rh1的抑制作用均比同浓度Rg1强.(3)Rh1与Rg1对LPS诱导的B淋巴细胞增殖均无明显作用.(4)不同浓度的Rh1与Rg1均能显著增强腹腔巨噬细胞吞噬能力(P＜0.01或P＜0.01),二者相比,Rh1的作用均比相应浓度的Rg1弱,且中、高浓度有显著差异(P＜0.01或P＜0.05).(5)Rh1在低、中浓度能促进腹腔巨噬细胞产生NO,Rg1在各个浓度均有此作用(P＜0.01或P＜0.01).结论:Rh1能够直接促进脾脏淋巴细胞增殖,能够下调ConA诱导的T淋巴细胞的增殖,且作用均比Rg1强,Rh1还能明显提高腹腔巨噬细胞吞噬能力,促进NO的释放,结果提示Rg1对机体的免疫调节作用可能是通过其肠内菌代谢产物Rh1入血后直接对T、B淋巴细胞和巨噬细胞作用的结果.
作者：蒋艳；王毅；邱全瑛 刊期： 2001年第08期
目的:内毒素是位于革兰氏阴性杆菌(G-)细胞壁外膜中具有高度生物学活性的以脂多糖(lipopolysaccharide, LPS)为主的成分.LBP(lipopolysaccharide-binding protein)/CD14(cluster of differentiation 14)系统在识别和调控LPS作用方面起着关键作用.甘氨酸是体内固有的简单的氨基酸,本教研室在先前的研究中发现甘氨酸对LPS的致热性有良好的拮抗作用,体外研究发现甘氨酸能破坏内毒素的构型,抑制内毒素诱导单核/巨噬细胞分泌TNF、IL-1等细胞因子.本研究是在前面研究的基础上进一步探讨甘氨酸对内毒素体内作用过程的影响,从而为研制有效的内毒素拮抗剂提供理论依据.方法:用RT-PCR技术检测大鼠肝组织LBP mRNA表达水平,并在此基础上观察甘氨酸对内毒素诱导大鼠肝组织LBP mRNA表达的影响.用原位杂交方法检测小鼠腹腔巨噬细胞CD14 mRNA表达水平,并在此基础上观察甘氨酸对内毒素诱导小鼠腹腔巨噬细胞CD14 mRNA表达的影响.结果:比较5组大鼠肝组织LBP mRNA的表达水平,内毒素组显著高于其他各组(P＜0.01),甘氨酸拮抗组显著低于内毒素组(P＜0.01),但甘氨酸拮抗组仍高于对照组.比较5组小鼠腹腔巨噬细胞的CD14 mRNA表达水平,内毒素组小鼠腹腔巨噬细胞CD14 mRNA表达水平显著高于其他各组(P＜0.01),甘氨酸拮抗组显著低于内毒素组(P＜0.01).结论:(1)内毒素可诱导大鼠肝组织LBP mRNA的表达.(2)甘氨酸可抑制内毒素诱导大鼠肝组织LBP mRNA的表达.(3)内毒素可诱导小鼠腹腔巨噬细胞CD14 mRNA的表达.(4)甘氨酸可抑制内毒素诱导小鼠腹腔巨噬细胞CD14 mRNA表达.
作者：单于；陆大祥 刊期： 2001年第08期
目的:采用免疫细胞化学方法探讨胃粘膜内CD3、S-100+树突细胞和nNOS的表达与慢性胃炎的关系及意义.方法:检测标本均取自胃窦部活检的胃粘膜组织.结果:CD3+细胞主要分布于上皮、腺上皮和固有膜内,而S-100+树突细胞则主要位于固有膜内,各组间细胞数量有显著差异(P＜0.01),nNOS阳性反应主要位于上皮和腺上皮的基底部,但各组之间nNOS的表达程度不同,特别是萎缩性胃炎与浅表性胃炎有显著差异(P＜0.01).结论:我们认为对CD3、S-100+树突细胞和nNOS的检测不仅有助于判断胃炎的病变程度,而且也为临床对胃炎的治疗提供新的启示.
作者：武一曼；葛振华；周凡；周维湛；唐福康；王若愚 刊期： 2001年第08期
目的:结核病疫情再度增高给结核病的防治提出了新的挑战,由于卡介苗对结核病的预防效果极不稳定,发展新型结核病疫苗势在必行,本研究通过对结核杆菌免疫保护性抗原Ag85A进行克隆与表达研究,旨在为结核病新型疫苗研制提供新的靶点.方法:根据结核杆菌H37Rv株免疫保护性抗原Ag85A的基因序列自行设计一对寡核苷酸引物,以结核杆菌H37Rv株基因组DNA为模板通过PCR扩增获得具有完整开架读码框(ORF)的Ag85A抗原基因,并将其正向插入真核和原核穿梭表达型载体pBKCMV构建重组质粒,重组质粒转入大肠杆菌后IPTG诱导表达,并对其表达产物进行Western-blotting分析.结果:①PCR扩增获得了具有完整开架读码框的Ag85A抗原基因;②构建了Ag85A抗原基因真核和原核穿梭表达型载体;③Ag85A抗原基因在大肠杆菌中实现了稳定表达.结论:本研究成功地对结核杆菌免疫保护性抗原Ag85A进行了基因克隆与表达,为进一步研究其在结核病新型疫苗研制中的应用奠定了基础.
作者：谢勇恩；鲍朗；胡昌华；张万江；陈玮 刊期： 2001年第08期
目的:建立结核杆菌检测基因芯片的制备技术.方法: 靶基因的制备、标记,用点样仪点于玻片介质上,并经过点样后处理制成基因芯片,用扫描仪测定处理前后芯片上荧光强度的变化,计算DNA固定率,分别设计了不同的玻片,不同的点液,不同的点样后处理方法的固定率的测定.结果:对结核杆菌检测基因芯片制备的条件和方法优化实验表明,用醛基修饰玻片作为介质,点样液用DMSO溶液作为点样液,点样后芯片处理,以水合1 h再干燥30 min为佳.结论:本研究建立的结核杆菌检测基因芯片的制备技术有较高的效率和可靠的实用性能.
作者：张万江；鲍朗；王晓樱；谢勇恩；陈玮；张会东；于向华 刊期： 2001年第08期
炎症反应是天然免疫的重要武器,内源性抗菌肽是天然免疫的重要介质.近5年来我们对哺乳动物抗菌肽进行了多方面研究,在此就某些有关实验研究作一概要综述.
作者：王伯瑶；黄宁；吴琦 刊期： 2001年第08期
目的:甲状腺素在正常能量代谢中有生热作用,是维持正常体温的因素之一.甲状腺机能低下时,代谢产热减少,体温也出现降低.所以临床上甲状腺机能低下的病人,常有喜热怕凉的症状.那么,在热环境中甲状腺机能低下时低体温是否可以恢复到正常水平本实验用抗甲状腺素药物,丙基硫氧嘧啶(PTU)复制甲状腺机能低下的大鼠在热环境中体温变化,来探讨甲状腺机能低下时引起代谢产热减少,是否是导致低体温的唯一原因.方法:实验分两部分完成.实验动物用SD大鼠.第一部分观察不同剂量的PTU对常体温环境下正常大鼠体温和血浆中T3、T4浓度的影响,实验分3组,每组8只动物,即对照组:自由饮自来水; PTU 10 mg组: 自由饮用PTU 10 mg/L自来水; PTU 30 mg组:自由饮用PTU 30 mg/L自来水.服用两周后,用数字体温计测量直肠温度,同时用放射免疫计数器测血浆中T3、T4浓度.第二部分分两组,每组8只动物,即对照组:自由饮用自来水;PTU组:自由饮用PTU 30 mg/L自来水.在服用两周后,先测直肠温度3次,然后将大鼠放入34℃热环境,再连续测直肠温度3次,同时测量血浆中T3、T4的浓度.结果:第一部分给大鼠连续服用两个不同剂量的PTU两周后,PTU组血浆中的T3、T4浓度和体温明显低于对照组,而且PTU 30 mg组体温更低(37.26℃±0.37℃,对照组37.91℃±0.29℃,P＜0.01).第二部分热环境(34℃环境)中,对照组的体温由实验前的(37.25±0.20)℃上升到(38.50±0.53)℃,平均升高1.25℃,PTU组由实验前(36.42±0.30)℃上升到(37.06±0.23)℃,只升高了0.64℃.结论:实验证明PTU引起大鼠甲状腺机能低下时体温也明显降低.但这种低体温可能不完全是由于代谢产热所致,也可能与甲状腺机能低下而影响体温调节中枢的功能有关.
作者：景志敏 刊期： 2001年第08期
目的和方法:吸烟是慢性阻塞性肺疾患、动脉粥样硬化等多种疾病的主要危险因素之一,但吸烟促使这些疾病发生的详细机制尚未完全明了.本文探讨作为香烟烟雾中主要成分的尼古丁对白细胞活化及白细胞与内皮细胞粘附的作用,并观察764-3(中药丹参提取物)对其影响,有助于阐明尼古丁在炎症性疾病发病中的作用并为寻找有效的防治炎症损伤的措施提供新的思路.结果:1.利用体外原代培养的人脐静脉内皮细胞(HUVEC)为靶细胞,首先观察到尼古丁促进中性粒细胞与内皮细胞粘附;2.用流式细胞术和Northern印迹杂交法分别观察到尼古丁可促进内皮细胞表面ICAM-1蛋白和mRNA表达,且呈时间和剂量依赖的趋势;3.用分子克隆技术成功地构建了4种含人ICAM-1基因不同长度的启动子序列和两种含顺式作用元件发生定点突变序列的荧光素酶报告基因质粒;4.用真核细胞转染技术证实尼古丁诱导ICAM-1基因表达依赖于其启动子中-230 bp和-134 bp之间的序列,且可能与此区域中的NF-κB结合位点有关;5.探讨了尼古丁的作用与细胞内信号转导机制之间的关系.观察到尼古丁可促使内皮细胞胞内游离钙离子浓度的增加(激光共聚焦显微镜法)、PKC活化(γ-32P-ATP掺入法)及Ras蛋白的活化(GST-RBD沉淀和Western印迹杂交法);6.观察到764-3可分别抑制尼古丁诱导的中性粒细胞与内皮细胞的粘附增加、内皮细胞ICAM-1蛋白和mRNA表达增加、内皮细胞胞浆游离钙离子浓度的增加以及PKC的活化.结论: 尼古丁可能通过作用于ICAM-1启动子区下游NF-κB位点,诱导ICAM-1基因表达,在尼古丁所致的炎症性疾病发病中起重要作用;764-3拮抗尼古丁的作用提示其在吸烟所致炎症性疾病的防治中可能具有潜在的意义.
作者：周卫辉；陈祥银 刊期： 2001年第08期
目的:从整体水平上探讨克雷伯氏肺炎杆菌内毒素诱导小鼠肺脏表达β-防御素的信号转导.方法:分别给予Toll受体蛋白-4基因发生点突变的小鼠C3H/HeJ及其野生型C3H/HeN腹腔注射LPS(4 mg/kg),于不同时间点采取气管、肺、肾等组织提取总RNA,用RT-PCR检测各组织β-防御素-3和/或β-防御素-4 mRNA表达,并对扩增出的cDNA片段进行测序;同步用Western blot法检测该两系小鼠肺脏I-κBα磷酸化状况(p-IκBα)和I-κBα的含量.结果:经LPS处理24 h后的C3H/HeN小鼠,其肺脏表达β-防御素-4 mRNA,而C3H/HeJ小鼠在相同条件下未见表达;与未给予LPS处理小鼠比较,经LPS处理4 h后,C3H/HeN小鼠肺组织p-IκBα含量明显增高;LPS处理后8 h,p-IκBα以及IκBα含量均呈减少趋势;至第24 h,p-IκBα和IκBα含量均明显减少,提示转录因子NF-κB活化.而在相同条件下,C3H/HeJ小鼠肺组织p-IκBα及IκBα含量均未见相应变化,提示克雷伯氏肺炎杆菌内毒素不能诱导TLR-4基因突变小鼠NF-κB和β-防御素基因的表达.结论:Toll/NF-κB信号转导通路介导克雷伯氏肺炎杆菌内毒素诱导小鼠肺脏表达β-防御素-4基因.
作者：蔡绍晖；陈新年；黄宁；王伯瑶 刊期： 2001年第08期
目的:循环中的肿瘤细胞必须牢固地粘附在血管壁上,才能进一步穿过血管壁,继续生长繁殖形成转移灶.本研究从肿瘤细胞与血管内皮细胞粘附的角度,研究活血化瘀药丹参注射液影响肿瘤转移的细胞及分子机制.方法: 以人脐静脉内皮细胞(HUVEC)为血管内皮细胞模型,药物及TNF(10×105 U/L)处理HUVEC 24 h后:1.倒置显微镜下观察细胞形态,研究丹参对TNF所致血管内皮损伤的保护作用;2.以蛋白染料染色法研究丹参对H-7402细胞与HUVEC粘附的作用(以A值表示);3.用细胞ELISA法观察HUVEC表面粘附分子ICAM-1的表达. 结果: 1.丹参对TNF所致血管内皮损伤有保护作用;2.TNF处理HUVEC 24 h后,H-7402与HUVEC的粘附A值为0.167±0.012,与对照(0.035±0.009)比,P＜0.01.若加入丹参0.125 mg、0.25 mg、1.25 mg、2.5 mg、12.5 mg,则粘附A值分别为:0.158±0.015、0.126±0.018、0.102±0.012、0.084±0.005、0.064±0.008,与TNF比,有显著差异,说明丹参可抑制TNF导致的H-7402与HUVEC粘附增加;3.TNF处理HUVEC 24 h后,HUVEC表面粘附分子ICAM-1表达的A值为0.238±0.008,与对照(0.087±0.009)比,P＜0.01.若加入丹参0.125 mg、0.25 mg、1.25 mg、2.5 mg、12.5 mg,则A值分别为0.209±0.016、0.194±0.011、0.175±0.013、0.145±0.006、0.108±0.014,与TNF比有显著差异,说明丹参可下调TNF诱导的HUVEC表面粘附分子ICAM-1表达的升高.结论: 丹参可通过下调血管内皮细胞粘附分子ICAM-1的表达及保护内皮细胞的完整结构而抑制肿瘤细胞与血管内皮细胞的粘附,从而抑制循环中的肿瘤细胞穿过血管壁,这可能是丹参抑制肿瘤血行转移的机制之一.
作者：郝钰；邱全瑛；吴君；方素萍 刊期： 2001年第08期
目的:探讨人中性粒细胞(PMNs)对人外周血单个核细胞(PBMCs)释放TNF-α的影响及其作用的初步机理.方法:采集健康供血者的新鲜外周静脉血,以葡聚糖沉淀和密度梯度离心法分离其PMNs和PBMCs,将PMNs与PBMCs按2∶1的数量比与脂多糖共同培育后,分别采用酶联免疫吸附法测定培养上清的TNF-α浓度,并用流式细胞测定结合荧光标记脂多糖的单核细胞的百分率及单核细胞表面平均荧光强度.结果:PMNs在细菌脂多糖刺激下不释放TNF-α,PMNs可以抑制PBMCs释放TNF-α,其抑制作用具有细胞特异性;经多聚甲醛固定的PMNs仍具有上述的抑制作用.流式细胞仪结果显示PMNs并不影响单核细胞与脂多糖结合.结论:PMNs可抑制人PBMCs释放TNF-α,其机理可能是PMNs干扰了脂多糖激活PBMCs的信号转导过程,抑制细菌脂多糖对其的活化,从而下调TNF-α的释放.
作者：李浩威；颜亮；潘剑波；杨皓庄；张穗梅；王彦平；付咏梅 刊期： 2001年第08期
本文对国外利用IL-1、IL-1R、ICE、IL-1ra,IL-1RacP,IL-6,IL-10, TNFR、cPLA2、COX及EP基金敲除小鼠研究细胞因子及前列腺素E2在不同致热原性发热机制的作用的研究概况进行了综述.局部感染或炎症如松节油(sc)性发热的机制较为简单,激活的信号转导途径也相当局限,依次由IL-1β、IL-6专一介导,其他细胞因子所起的作用甚微;全身感染/炎症如细菌脂多糖(LPS,ip)诱导发热的机制较为复杂,其中小剂量LPS性发热依赖于IL-6,IL-1可能参与其过程,两者的表达及作用均受到IL-10的调节;大剂量LPS引起的发热则激活较为广泛而复杂的体温调节信号通路,其中包括体温的负向调节和正向调节机制,IL-6依赖性和非依赖性通路,但以后者为主,且这些通道存在复杂的相互诱导、代偿或抑制,其作用因剂量而异.IL-1、TNF及IL-10均参与其机制,其中TNFα和IL-10的作用形式异常复杂,可因剂量差异完全相反.病毒对体温的影响也较为复杂,似乎有IL-1β的参与,并由IL-6介导.上述发热的诱导皆需要AA→PGE2级联反应系统的参与,非PGE2依赖性信号转导系统对之均贡献甚微,但物理性应激反应引起的体温升高似乎例外.
作者：李沧海；霍海如；姜廷良 刊期： 2001年第08期
目的:旨在观察α-黑色素细胞刺激素(α-melanocyte stimulating hormone, α-MSH)对LPS部分生物学活性的影响,进一步探讨α-MSH的抗炎作用及免疫调节功能.方法:应用比色法、倒置生物显微镜及流式细胞仪,测定在LPS作用下α-MSH对小鼠腹腔巨噬细胞释放H2O2量、中性粒细胞凋亡率及FITC-LPS与单核细胞的结合率及单核细胞表面平均荧光强度的影响.结果:LPS可刺激小鼠腹腔巨噬细胞释放H2O2,而α-MSH与LPS共同培养,则能明显抑制巨噬细胞释放H2O2(P＜0.01);α-MSH及LPS本身均不影响中性粒细胞凋亡(P＞0.05),但在LPS作用下,α-MSH可显著促进中性粒细胞凋亡(P＜0.01);并且,α-MSH可降低FITC-LPS与单核细胞的结合率及单核细胞表面的平均荧光强度(P＜0.05,P＜0.01).结论:以上结果表明,α-MSH不仅能有效抑制LPS刺激巨噬细胞释放H2O2、促进LPS作用下的中性粒细胞发生凋亡;而且可干扰LPS与单核细胞的结合,发挥其有效的免疫调控作用,对控制局部和全身炎症反应具有重要意义.
作者：陈波；胡巢凤；王彦平；陆大祥；颜亮；戚仁斌；张穗梅；付咏梅；李楚杰 刊期： 2001年第08期
目的和方法:中药的低毒性在肿瘤的预防研究中日益受到人们的重视.本研究用具有益气补肾、利湿解毒和活血散结作用的桷斗芪散处理动物,采用Solt-Farber的AFB1致大鼠肝癌前病变的短期实验模型,以大鼠肝癌前病变灶中NO浓度为实验指标,观察中药方剂在防治大鼠肝脏癌变过程中对NO产生的影响.Solt-Farber模型能有效地引起肝脏产生许多变异肝细胞灶、增生结节,其中一部分在致癌剂和促癌因素影响下将进一步发展为癌前病变及癌.此模型的实验程序包括3个部分:注射AFB1,进食含2-AAF的饲料加上PH.用试剂盒检测肝组织中的NO浓度.结果和结论:肝癌前病变灶中NO浓度明显低于正常对照组,而用桷斗芪散处理后,病变灶中NO浓度明显增高,肝内阳性病变灶减少.说明桷斗芪散可抑制肝癌前病变灶的增生,其机理之一可能是通过提高局部NO浓度而起作用的.NO是一种简单而不稳定的自由基气体,很多细胞均有合成NO的酶,NO可作为介质、信使或细胞功能调节因子,参与机体许多生理和病理过程,其与肿瘤生物学的关系以及在肿瘤免疫中的作用亦受到关注.关于桷斗芪散调节NO产生的机理尚需进一步探讨.
作者：王学江；钱英；丰平；文朝阳；姜大巍 刊期： 2001年第08期
目的:观察人血浆高密度脂蛋白(HDL)对大鼠内毒素血症的治疗和预防效果.方法:实验分为对照组、治疗组和预防组.对照组(n=10):仅静脉输注内毒素(ET,500 EU/kg);治疗组(n=8):先静脉输注ET,待血压明显下降及ET输注后1 h,再静脉输注HDL(75 mg/kg);预防组(n=8):输注HDL后再输注ET.分别在3个不同时点,即第1时点(输注ET或HDL前,为自身正常对照)、第2时点(ET输注后1 h)和第3时点(ET输注后2 h)静脉采血,采用鲎试剂法和放射免疫分析法测定大鼠血浆ET水平和肿瘤坏死因子(TNF)浓度并观察血压及存活时间.结果: (1)输注ET后对照组大鼠血压进行性下降(P＜0.01);输注HDL后治疗组大鼠血压下降程度明显弱于对照组(P＜0.01);预防组大鼠输注ET后其血压无明显下降(P＞0.05).治疗组及预防组大鼠存活时间均较对照组明显延长(P＜0.01).(2)3组大鼠血浆ET水平在各时点均无明显差异(P＞0.05).(3)无论是治疗组还是预防组,其血浆TNFα水平于第3时点均明显降低(P＜0.05).结论:人血浆HDL能减轻或抑制内毒素血症大鼠血压的下降并明显延长其存活时间,说明人血浆HDL能提高机体对抗ET损伤的能力,对内毒素血症具有较理想的治疗和预防作用;人血浆HDL拮抗ET损伤的机理可能与其抑制TNF释放有关.
作者：黄英；顾玲；王树人；康焰；吴淑红 刊期： 2001年第08期
目的:探讨雷公藤多甙对小鼠移植物抗宿主病的作用.方法:供鼠BALB/C,雌性,受鼠C57BL/6,雄性.无菌取供鼠脾脏及双侧股骨骨髓,制成混合细胞悬液.受鼠在骨髓移植前24 h内应用直线加速器全身照射,总剂量为1 800 cGy,剂量率为200 Gy/min,将制成的混合细胞悬液经尾静脉输给受鼠0.5 mL,含骨髓细胞1×106个,脾淋巴细胞1×107个,制成小鼠aGVHD模型.随机分成5组:同基因移植组,异基因移植组,CsA+MTX组,GTT组,GTT+CsA组.CsA+MTX组受鼠骨髓移植后第1-10 d,每日腹腔注射CsA2.5 mg*kg-1*d-1, 第1、3、5、9 d腹腔注射氨甲蝶呤(7 mg/M2).GTT组受鼠骨髓移植后第1-10 d,每日腹腔注射GTT 5 mg*kg-1*d-1.GTT+CsA组受鼠骨髓移植后第1-10 d,每日腹腔注射GTT2.5 mg*kg-1*d-1、CsA2.5 mg*kg-1*d-1,第11 d处死各组受鼠,用ELISA法检测血清中细胞因子IL-2、TNF-α、IL-4、IL-10的浓度.用流式细胞仪检测脾T淋巴细胞及表面粘附分子的阳性表达率.结果:(1)雷公藤多甙组小鼠11 d生存率明显高于异基因移植组、CsA+MTX组.(2)雷公藤多甙组小鼠CD+3、CD+4、CD+8、CD+4CD11a+、CD+4CD18+、CD+8CD11a+、CD+8CD18+细胞的百分率明显低于异基因移植组.(3)雷公藤多甙组小鼠血清TNFα、IL-2浓度低于、IL-10高于异基因移植组.结论:(1)雷公藤多甙可明显提高aGVHD小鼠的生存率.(2)其机制与:①雷公藤多甙可降低T细胞及亚群数量及其相关粘附分子的表达有关.②雷公藤多甙降低促进aGVHD的细胞因子-TNFα、IL-2的表达,提高抑制aGVHD的细胞因子-IL-10的表达有关.
作者：于艳秋；张海鹏；包娜仁 刊期： 2001年第08期
目的:糖尿病肾病(DN)作为糖尿病(DM)并发症,本质上是DM血管损伤的表现.DM早期肾脏肾小球高滤过是DM血管损伤始动因素,导致血管通透性增加,前列腺素系统代谢失常参与了这一机制的形成,DM脂质过氧化加重DM血管损伤的进程.有研究表明血浆脂质过氧化损伤与TXA2/PGI2失调在糖尿病性血管病变发生中起重要作用,本实验拟观察早期实验性DM大鼠前列腺系统代谢失常和氧化损伤在DN发病中的作用,通过止消通脉宁的治疗作用,探讨中药复方改善糖尿病肾脏病变的机制.方法:采用链脲佐菌素(STZ)造成糖尿病大鼠模型,分正常对照组,模型组,中药治疗组:成模后给予止消通脉宁醇提剂(由黄芪、生地、鬼箭羽、大黄等组成每毫升含生药2 g)按60 mg*kg-1*d-1灌胃;西药治疗组:成模后给予糖适平5 mg*kg-1*d-1、洛丁新15 mg*kg-1*d-1;灌胃.每组10只,以上各组均予普通饲料,自由饮水,实验周期为2周、4周.测定尿微量白蛋白(m-Alb)、内生肌酐清除率(Ccr)、肾重/体重、肾组织丙二醛(MDA)含量、血浆6-keto-PGF1α和TXB2比值.结果:各组DM大鼠无论14 d或28 d尿m-Alb均明显升高,与正常对照组比有显著差异(P＜0.01); 中、西药治疗组较模型组明显减少(P＜0.01),但中、西药治疗组未见差异(P＞0.05).各组DM大鼠肾重/体重增加,Ccr显著升高(P＜0.01),中、西药治疗组肾重/体重、Ccr显著低于模型组,中、西药治疗组6-keto-PGF1α/TXB2比值明显降低,MDA含量亦明显降低.讨论和结论:本研究结果表明,模型组血浆TXB2/6-keto-PGF1α比值升高,6-keto-PGF1α水平降低,与相关文献结果一致.经止消通脉宁治疗后,TXB2含量和TXB2/6-keto-PGF1α比值明显降低.提示止消通脉宁能够改善糖尿病大鼠血浆TXB2与PGI2间平衡,以抑制血小板活化,防止血管内皮细胞损伤和血栓形成及降低肾小球高滤过;本实验以Ccr代表GFR;肾组织MDA含量代表产生LPO量,判断是否发生脂质过氧化反应.结果说明,DM时,GFR增加,Ccr明显高于正常对照组;肾组织MDA含量均升高,提示PG代谢紊乱、脂质过氧化反应可能参与了肾脏肥大、高滤过现象.止消通脉宁治疗后MDA含量明显降低,表明止消通脉宁能提高肾脏功能减轻体内自由基代谢紊乱状态.止消通脉宁能减轻自由基代谢紊乱可能与方剂中所含中药具有清除自由基和(或)抗氧化作用有关,据报道黄芪有抗氧化作用,大黄能抑制DN肾脏肥大及高滤过,改善脂质代谢紊乱,这些中药均是止消通脉宁的主要成分.另外,如鬼箭羽、大黄等活血化瘀中药能纠正组织自由基代谢紊乱,诸药配伍对降低肾脏脂质过氧化物形成,改善糖尿病肾脏病变起重要作用.
作者：赵雁；王耀献；黄启福；吕仁和 刊期： 2001年第08期
目的与方法:采用放免法及下丘脑中PGE2含量及COX活性进行同步检测,以期探明桂枝汤对体温双向调节作用与下丘脑组织中PGE2含量及COX活性的关系.结果:在酵母诱导发热大鼠中,桂枝汤对大鼠体温升高有显著的降低作用,在安痛定诱致的低体温大鼠中,桂枝汤对大鼠体温降低有显著的回升作用,给药后体温及体温变化差值与模型组比较均有显著差异,表明桂枝汤可使高、低体温动物分别向正常水平方向进行调节.在对不同体温状态大鼠发挥解热或抗低温作用的同时,桂枝汤对酶母诱导发热大鼠下丘脑中异常增高的PGE2含量有显著的降低作用,而对安痛定诱致的低体温大鼠下丘脑中异常降低的PGE2含量又有显著升高的作用.但桂枝汤对两种模型大鼠下丘脑细胞中COX活性影响不明显.结论:桂枝汤双向调节体温的作用机理可能部分是通过影响下丘脑组织中PGE2含量来实现的.但PGE2含量增减不依赖于下丘脑细胞中的COX,由于COX是PGE2生成中重要的限速酶,因此我们认为下丘脑中刺激体温变化的PGE2,来自自分泌(下丘脑细胞自身合成后分泌至胞外,再刺激分泌细胞)的可能性相对较小.
作者：齐云；李沧海；郭淑英；周军；霍海如；姜廷良 刊期： 2001年第08期
目的:观察桂枝汤对模型动物退变颈椎间盘组织磷脂酶A2(PLA2)活性的影响,借以探讨桂枝汤治疗颈椎病的抗炎作用机理.方法:选用动力失衡性颈椎病动物模型,造模动物到期后取C4-5颈椎间盘,采用催化活性与微孔比色相结合的测定法测定PLA2活性.结果:模型组颈椎间盘PLA2活性明显升高,正常组则很低,二者比较显著差异;桂枝汤无论是对早期治疗组还是治疗组均有下调PLA2的作用,以大剂量为明显.芬必得同样表现出明显下调PLA2活性的作用,与桂枝汤比较无显著差异.结论:桂枝汤具有下调退变颈椎间盘中PLA2活性的作用,从而减少多种炎症介质的合成,这可能是其治疗颈椎病的作用机制之一.
作者：周军；方素萍；霍海如；齐云；姜廷良 刊期： 2001年第08期
作者：戚仁斌；陆大祥；王华东；王海华；颜亮；王彦平；付咏梅；李楚杰 刊期： 2001年第08期
