AIM To construct an expression vector for anti-HBsAg antibody Fab fragment and interferon-aA (IFN-aA)fusion protein in E. coli.METHODS With PCR and molecular clone techniques, we amplified the gene fragment of IFN-aA withcorresponding endonuclease sites and artificial linker at 5', 3' termini, and then formed pHS/IFN-aA byrecombining it within the vector in correct endonuclease sites, choosing the positive clone to transform intoE. coli and intoduced by IPTG to express the fusion protein.RESULTS Enzymic hydrolysis and DNA sequence measurement confirmed that human gene of IFN-aA wascorrectly cloned to the vector and could express fusion protein in E. coli.CONCLUSION The success in construction and expression of a fusion protein makes it possible to carry outfurther studies on its purification and targeted polypeptide therapy to HB virus.
作者: 刊期: 2000年第03期
AIM To observe the effect of repair of intrathoracic esophageal perforation with gastric seromuscularpatch.METHODS Twelve patients with intrathoracic esophageal perforation, none of whom were associated withcarcinoma, underwent primary repair with a gastric seromuscular patch and omental pedicle flap between1996 and 1999. Four perforations were iatrogenic and 8 were spontaneous. The interval from perforation tooperation was less than 12 hours in 4 patients, 12 to 24 hours in 3, and more than 24 hours in 5. The principleof repair included: ① To expose health mucosa and submucosa for the primary repair. The leading edge ofthe mucosa is grasped and the adjacent esophageal muscle is mobilized away from the submucosa until there isa 3 to 7 mm circumferential rim of normal submucosa. ② The gastric seromucular patch and omental pedicleflap require continuity of the right gastric piple, omental vessels and the rami. ③ To assure the blood flow ofthe esophagus when the necrotic mediastinal and esophageal tissue are debrided, the normal esophageal tissueoften extends well beyond the 3/4 diameter of the esophagus. ④ The mediastinal pleural is not closed inorder to drain. The catchers nasogastric tube decompression of the stomach is continued until thepostoperative ileus resolves.RESULTS Eleven of the 12 patients underwent primary repair, one patient had leak at the site of repairand died.CONCLUSION Meticulous repair of an intrathoracic esophageal perforation using a gastric seromuscularpatch is the preferred approach regardless of the duration of the injury.
作者: 刊期: 2000年第03期
The current established drugs used to treat inflammatory bowel disease include glucocorticoids includingnewer agent budesonide, sulfasalazine and 5-ASA compounds such as Asacol, Pentasa, Dipentum andBalsalazide and immunomodulatory agents such as azathioprine, and 6-mercaptopurine. Additional drugswhich have been found to be useful, particularly in refractory cases of Crohn's disease including fistulizingtype of Crohn's disease, include cyclosporine A, methotrexate, humanized antibody against TNFa(cA2),FK506, IL-10, IL-11 and Probiotics. Various agents, whether used alone or in combination, have to betailored for each patient and none is ideal. Exciting new developments directed against proinflammatorypathways, cytokines, free oxygen radicals and cell surface related immune targets are areas of intense recentinvestigations and many novel therapeutic agents are expected to be available in the near future for medicaltreatment of inflammatory bowel disease.
作者: 刊期: 2000年第03期
AIM To explore the method to prolong the survival of patients with postoperative rectal cancer and toprevent its recurrence.METHODS To analyze the reasons of recurrence after curative resections for rectal cancer of 399 patients.RESULTS Ninety out of 399 patients who received curative excision died of relapse of the tumor. Pelvicrecurrence was found in most of the patients followed by liver and pulmonany metastasis. The survival timein Dukes A was the longest and in Dukes C the shortest. The survival period was 12 months longer, in thepatients receiving pre-operative radiotherapy than that did not. Inadequate excision of the primary tumor orthe draining lymph nodes was the main cause for local recurrence.CONCLUSION Adequate surgery and adjuvant therapy are the most effective methods to prolong thesurvival of patients with postoperative rectal cancer and to prevent its recurrence.
作者: 刊期: 2000年第03期
AIM To characterize genomic DNA flanking IS900 insertions and develop a rapid Multiplex PCR IS900Locus (MPIL) typing method for MAP reporting the presence or absence of the element at each locus,METHODS Genomic DNA flanking 14 of the 18 IS900 loci was sequenced and compared with databasehomologues. An MPIL typing method was developed using a common IS900 primer and individual locus-specific primers designed to produce amplification products differing by about 50bp which could be easilyresolved on a single gel. MPIL was applied to a panel of 81 MAP isolates and compared with RFLP profiles.RESULTS Genes flanking IS900 loci included homologues of transcription regulators, a sigma factor, anitrate reductase, a polyketide synthase and an O6-methylguanine-methyl transferase. MPIL rapidly andconsistently identified 10 individual types of MAP from the panel of 81 isolates, and distinguished betweenbovine and ovine strains. Nine MPIL types corresponded directly to single RFLP types previously identified.CONCLUSION Isg00 insertions in MAP may affect the expression of genes critically associated with thepathogenic phenotype. MPIL typing can identify bovine and ovine strains independent of the need for cultureand may contribute to studies of the molecular epidemiology of these difficult organisms.
作者: 刊期: 2000年第03期
AIM To provide evidence that UBT is the most cost-effective tool for evaluation of H. pylori eradication.METHODS Data on twenty-six consecutive patients at Atlanta VA Hospital who underwent UBT wereretrospectively reviewed. All patients had endoscopic diagnosis of peptic ulcers and biopsy proven H. pyloriinfection. Eight to ten weeks after completion of triple therapy (amoxicillin, biaxin and prilosec), allpatients had C14 UBT (PY test kit, Charlottesville, Virginia). Ten patients had repeated endoscopicexaminations and gastric biopsies. Twelve patients had serology tests for H. pylori.RESULTS UBT was negative in all patients (two patients had indeterminate result on the first time, butshown to be negative on the second UBT). Biopsies from all ten patients who were re-endoscoped werenegative for H. pylori. Serology tests on all the twelve patients were positive. Cure of H. Pylori could notbe determined on the titer change. All patients spent about 30 minutes in nuclear medicine laboratory, theendoscopic patients spent 2 hours to 4 hours in endoscopic laboratory; the cost for a UBT was about 50, thecost for an endoscopy with biopsy was above 200.CONCLUSION The 04 UBT is a rapid, economic and accurate test to monitor H. pylori eradication. Thetest should be considered a gold standard test for evaluating the effectiveness of treatment of H. pyloriinfection, unless patients need repeated endoscopy to rule out gastric cancer.
作者: 刊期: 2000年第03期
AIM The incision in rectal cancer operation is adopted commonly in the left mid-lower abdomen. But thereare some defects for the incision, which is close to the artificial colotomy, readily be contaminated by feces,difficult to treat the lesions in hepatic and cholecystic area at the same time and in the weakened area ofabdominal wall. So, we employed the abdominal right lower paramedian incision to solve these problems.METHODS The abdominal right lower paramedian incision is from publc tubercle upward to 3 cm- 4 cmabove navel. The incision should be extended upward if individual need of performing hepatic and cholecysticoperation, or placing catheter or pump in hepatic artery or portal vein for chemotherapy at the same time.RESULTS One hundred and eighty three cases with rectal cancer were adopted this incision in differentoperation procedure, and out of them 41 patients were taken different operation on hepatic and cholecysticlesions and place a catheter or pump to hepatic artery or portal vein. Operators feel that the incision dose nothinder exploring and operating in all of the patients.CONCLUSION The right lower paramedian incision of abdomen is far away from the artificial colotomy,and it can reduce the feces contamination, lower down the rate of incision hernia and paramedian hernia orfistula. Furthermore, it is easy to treat the complicated hepatic and cholecystic lesion. So, authors suggestthat this incision is useful for the operation of rectal cancer, and it is worth to populize in clinical practice.
作者: 刊期: 2000年第03期
AIM To evaluate a system for bile acid formation in human hepatocytes in comparison with HepG2 cells.METHODS Hepatocytes were isolated from normal human liver tissue and were cultured in serum-freeWilliam's E medium. The medium was collected and renewed every 24 h. Bile acids and their precursors inmedia were finally analysed by gas chromatography-mass spectrometry.RESULTS Cholic acid (CA) and chenodeoxycholic acid (CDCA) conjugated with glycine or taurineaccounted for 70% and 25% of total steroids. One third of CDCA was also conjugated with sulphuric acid.Dexamethasone and thyroid hormone alone or in combination did not significantly affect bile acid formation.The addition of cyclosporin A (10 tm) inhibited the synthesis of CA and CDCA by about 13% and 30%,respectively.CONCLUSION Isolated human hepatocytes in primary culture behave as in the intact liver by convertingalmost quantitatively cholesterol to conjugated CA and CDCA. This is in contrast to cultured HepG2 cells,which release large amounts of bile acid precursors and unconjugated bile acids into the medium.
作者: 刊期: 2000年第03期
AIM To determine NO, NO synthase (NOS) and NOSmRNA of the esophageal carcinoma cells (SHEEC1)in apoptotic process induced by As2O3 and to explore the relationship between NO and apoptosis.METHODS The apoptosis of the cell line (SHEEC1) was induced by arsenite (As2O3, 5 μmol/L and10 μmol/L). In the process, at 2 h, 4 h, 8 h, 16 h and 24 h after administration of As2O3, NO production incultural medium was detected quantitatively by spectrophotometry; NOS Ⅱ was detected byimmunohistochemistry and NOS mRNA by in situ hybridization (ISH). The cells at endpoint of theexperiment were examined under transmitted electron microscope (TEM) for apoptosis.RESULTS The amount of NO released from SHEEC1 were increased from the basal condition (0.68×10-2μmol/L) up to the high level (2.38×10-2μmol/L) at h 16. The increment of NOS Ⅱ was found afteradministration of As2O3; the intracytoplasmic ISH signals of NOSmRNA in small size was found firstly at4 h, and then became highly predominant. Apoptotic changes of SHEEC1 occurred at 24 h under TEM.CONCLUSION After administration of As2O3, NO released from cultured SHEEC1 cells was detected withincreasing amount up to 16 h. The expression of NOS H and transcription of NOSmRNA are upregulated.The present findings suggest a concept that the NO may be a mediated and effective factor in apoptosisinduced by As2O3,
作者: 刊期: 2000年第03期
AIM To probe into the feasibility and theoretic basis for the treatment of chronic hepatitis with hyperbaricoxygenation (HBO).METHODS Sixty cases of chronic hepatitis were randomly distributed into an experimental group (n=30)and a control group (n =30). The experimental group was treated with HBO for 6 courses. The controlgroup was treated with commonly used drugs in clinic for 60 days. The function and blood stream graph ofliver were examined and the liver biopsies were made before and after treatment. The routine paraffin slidesof liver tissue were cut, stained with HE, and observed under optical microscope. The ultrathin slides fromparaformaldehyde and glutaraldehyde fixed liver tissue were cut, stained with lead citrate, and observedunder transmission electric microscope. The HBsAg and HBcAg in the experimental group were detected bythe ABC immunohistochemical method before and after treatment.RESULTS In the experimental group the ALT, SB, γ-GT, AKP, IgG and IgM in blood (P< 0.05) and thedegeneration and necrosis of hepatocytes (P<0.05) were remarkably decreased, the mean contractive waveof blood stream in the liver (P<0.05) and the blood stream in the right ramus of janitrix (P<0.05) weresignificantly increased, and the swelling of mitochondria, the increased lysosomes, the generation of Kupffer's cells, the infiltration of lymphocytes in portal area and the capillary generation were remarkably alleviated(P<0.05), but the fibrosis and fat-storing cells did not reduce (P>0.05) in the liver, and the expression ofHBsAg and HBcAg in the liver was not lowered (P<0.05) after the treatment with HBO.CONCLUSION The treatment with HBO for chronic hepatitis was effective and recommendable.
作者: 刊期: 2000年第03期
AIM To observe the therapeutic effect of colloidal bismuth tartrate in an animal colitis model.METHODS Immune-complex colitis was induced in groups of rabbits by formalin, and two hours later0.85 mL heat-aggregated rabbit IgG was given intravenously through the ear cannula. Animals wereintracolonically treated with colloidal bismuth tartrate (BITNAL), and its effect was compared withsulfasalazine (SASP), indomethacin (IND) and bifidobiogen (BIFG). Animals were killed, the mucosalappearance was scored (0-4), and tissue saved for histological studies, the number of neutrophils present ininflamed colonic tissue was quantitated by the myeloperoxidase (MPO) activity assay, the production oflipoxygenase and cyclo-oxygenase products was monitored and eicosanoid production were assayed byincubation colonic specimens and the media for prostaglandin E2(PGE2), leukotriene (LTB4), thromboxaneB2(TXPe) were examined by radiommunoassay.RESULTS Immune-complex colitis was induced by formalin and IgG, colonic damage persisted for at least1 wk by macrography. Histologically, the inflammatory response included mucosal and submucosalinfiltration by polymorphonuclear leukocytes, macrophages, lymphocytes and fibroblasts, the macroscopic,persent 2 wk after IgG, was correlated with greatly increased PGE2, LTB4 and TXB2 compared with levels incontrols. Treatment with BITNAL (500 mg/kg) resulted in a lowered inflammation index, lowered MPOactivity and inhibited the increased formation of PGF-2, LTB4 and TXB2 by the inflamed colon, and IND(500 mg/kg) markedly inhibited prostanoid formation in both inflamed and control colon but did not reducetissue damage, SASP (500 mg/kg) also inhibited the formation of PGE2, LTB4 and TXB2 but the effectswere less marked. BIFG (400 mg/kg) did not significantly reduce the colonic injury and the media sythesizedby the rabbit colon.CONCLUSION BITAL provides better therapeutic effects in experimental colitis than anti-inflammatorydrug IND or SASP.
作者: 刊期: 2000年第03期
AIM To study the clinical significance of alterations of serum hyaluronic acid in rats with carbontetrachioride-induced liver fibrosis.METHODS Rat liver fibrosis model was induced by carbon tetrachloride (CC14). The rats were divided intofive groups; group 1 (control): 0 week with no CCl4-inducing; group 2, 3, 4 and 5: 3, 6, 9 and 12 weeksafter CCl4-induction respectively. Serum HA level was analysed among various liver fibrosis groups andcontrol, and then compared the HA findings with the hepatic histopathology.RESULTS During rat liver fibrosis, serum HA levels of the liver fibrosis groups (group 2: 7.98ng/mL;group 3: 20.10 ng/mL; group 4:229.73 ng/mL; group 5:324,74 ng/mL) were significantly higher thanthat of control group (group 1:0.21 ng/mL) (P<0.01), in which group 4 and group 5 are much higher1094 times (229.73ng/mL/0.21 ng/mL) and 1546 times (324.74 ng/mL/0.21 ng/mL) than group 1respectively. When compared with each other, the serum HA levels are 38 times (7.98ng/mL/0.21 ng/mL; P<0.01, group 2 vs group 1); 2.5 times (20.10ng/mL/7.98 ng/mL; P<0.01, group 3 vsgroup 2); 11.4 times (229.73 ng/mL/20.10 ng/mL; P<0.01, group 4 vs group 3); 1.4 times (324.74 ng/mL/229.73 ng/mL; P<0.01, group 5 vs group 4) respectively.CONCLUSION The results demonstrated that the dynamic alterations of serum HA play an important rolein the early clinical diagnosis and staging of liver cirrhosis.
作者: 刊期: 2000年第03期
AIM To investigate the protective effects and mechanism of Diltiazem (Dil) on liver, pancreas and smallintestine in hemorrhagic-shock canine.METHODS The canines were bled to a mean arterial pressure (MAP) of 5.33kPa-6.67kPa for 30min toestablish the shock model. During the shock state, the dogs received either water-soluble calcium blocker Dilor saline solution. The MAP was kept at this level for 90min, then the total blood which was bled previouslywas reperfused. The total observation time of the experiment was 240min.RESULTS Dil could significantly increase MAP from 150min to 240min (P<0.01) and the activity ofsuperoxide dismutase (SOD) of pancreas tissue (P<0.01), and it could also decrease the content ofmalondialdehyde (MDA) in liver, pancreas and small intestine tissues (P< 0.01) and the activity of SOD ofthe liver and small intestinal tissues (P<0.01) in the canines. Electron microscopic data indicated that theultrastructures of liver, pancreas and small intestine tissues were normal in Dil group.CONCLUSION Dil can protect the structure and function of the liver, pancreas and small intestinal inhemorrhagic-shock canine.
作者: 刊期: 2000年第03期
AIM To understand the pollution status of drinking water in Cixian area and to find out a way to solute it.METHODS Contents of nitrate nitrogen, nitrite nitrogen and ammonia nitrogen were measured andcompared with controls and national standards.RESULTS Levels of nitrate nitrogen, nitrite nitrogen and ammonia nitrogen were higher than the nationalpermissive levels by 20.60%, 50.50% and 33.30% respectively. Also the levels in Cixian are significantlyhigher than those of a controls areas of Chicheng - a county with very low incidence rate of esophagealcancer.CONCLUSION Three nitrogen levels in drinking water correlate both with the amount of N-containingfertilizer used in these areas and the mortality rates of esophageal cancer.
作者: 刊期: 2000年第03期
AIM We introduce a new highly selective vagotomy (HSV) procedure which can completely destroy anyexisting variant vagal fibers innervating the proximal stomach.METHODS Twenty-eight dogs were divided into three groups: group A had retrograde liberated highlyselective vagotomy (RLHSV), group B had traditional HSV and group C as control group. Gastric acidsecretion function was tested and gastric parasympathetic and sympathetic enervation were studied byretrograde tracing the horseradish peroxidase (HRP) or by histochemical staining. After being convincedthat the modified procedure was an easier and more effective HSV on animal model, we applied thetechnique to the treatment of duodenal ulcers in 100 patients.RESULTS Significant decrease of acid secretion was found in operated dogs in both group A and B. After injection of HRP subserously into the fundus and body, no HRP positive cells were found in the vagal dorsalnucleus in group A and B but were in group C. When HRP was injected into the antrum, HRP stained cellswere found in dorsal nucleus in all three groups. The negative histochemical staining of AchE and the positivefluorescence in the fundus and body in group A and B indicate that parasympathetic nerve fibers weredestroyed completely while norepinephrinergic nerve fibers were intact. When the operation was applied topatients, no severe complications or operative mortality occurred. Patients were followed up for 6- 84months. They all had complete gastric emptying function. Ninety-four patients were graded as Visick I orⅡ. Six as Visick Ⅲ and Ⅳ. Eighty patients had been followed up with gastroscopy for two years afteroperation. Four patients had recurrent ulcers. Among them two patients were asymptomatic and the othertwo had symptoms and were treated with partial gastrectomy.CONCLUSION Compared to the classic HSV, our modified procedure is more effective clinically andconvenient technically.
作者: 刊期: 2000年第03期
AIM To observe the therapeutic effect of weile jiaonang (WLJN) for peptic ulcer (PU) and its toxicity toanimals, and to find out the nontoxic Chinese medicines in replacing those chemical medicines with sideeffect.METHODS Five hundred and forty patients with PU were divided into three groups, therapeutic group byWLJN, control group taking famotidine and combined group with WLJN and famotidine. One hundred andeighty patients in each group were studied. The diagnosis of 540 patients with PU is in accordance with thebasis provided by the National Scientific Congress of Digestive System Disease convened in Hangzhou, 1978.The study followed the criterion provided by this Congress.RESULTS Total effective rate in therapeutic group was 93.3%, and 93.3% in control group. Combinedgroup achieved a total effective rate of 100%. For all three groups, there was no significant differencestatistically, but the control group appeared obvious side effects. The result of acute toxic experiments onwhite mice showed that there no death and toxic side effect even the highest dosage was given, which wasequivalant to 120 times to the dosage of patients. The result of chronic toxic experiment showed there was noobvious pathologic change in three dosed groups. The tails of mice in dosed groups were more smooth delicateand cleaner than those in control group.CONCLUSION Through observation in clinic and exprimental animal, it is proved that weilejiaonang isparticularly effective and safe Chinese patent drug against PU.
作者: 刊期: 2000年第03期
AIM To evaluate the clinical value of creatine kinase macroisoenzyme type 2 (CK-M2) and oligosaccharideprotein (OP) in serum from patients with gastric carcinoma (GC).METHODS Serum level of CK-M2 was detected by agar gel electrophoresis. OP concentration was measuredby an enzyme immunoassay.RESULTS Serum levels of CK-M2 and OP in 57 cases of GC were significantly higher than those in 51 caseswith gastric precancerous lesion and 28 controls. The diagnostic sensitivity and specificity for GC with CK-M2 was 56.10% and 98.63% respectively. CK-M2 and OP were not associated with histologic type and degreeof differentiation.CONCLUSION These results suggest that CK-M2 may serve as a marker to diagnose GC, and the specificityis higher, whereas OP is not more significant for GC diagnosis, but it could be a useful indicator forevaluation the status of body immune.
作者: 刊期: 2000年第03期
AIM To study the combined expression of gastrointestinal hormone substance P and anti-apoptosis gene Bcl-2 in gastric carcinoma and its significance.METHODS Substance P and Bcl-2 protein expression was examined by the S-P immunohistochemicalmethod in 33 cases of gastric carcinoma, 17 adjacent the carcinoma and 13 normal gastric mucoma.RESULTS Positive expression of SP in gastric carcinoma was higher than that of both adjacent and normalmucosa (P < 0.001). There was no statistical difference in the positive expression between adjacent andnormal mucosa (P > 0.05). The expression of bcl-2 both in gastric carcinoma and adjacent tissues werehigher than that of normal gastric mucosa (P< 0.05-0.01). But the positive expression of Bcl-2 had nostatistical significance between gastric carcinoma and adjacent tissues.CONCLUSION Both gastrointestinal hormone SP and Bcl-2 gene have synergistic expression in gastriccarcinoma, indicating that they all take part in the occurrence of gastric carcinoma. Abnormal expression ofBcl-2 gene occurred in benign gastric pathological changes, once they become carcinoma, the positiveexpression of cell is no more increased, possibly because that there is no more increase of the intensity of Bcl-2 inhibition of cell apoptosis.
作者: 刊期: 2000年第03期
AIM To investigate the effect of boschniakia rossica (BR) extract on expression of GST-P, p53 and p21fasproteins in early-stage chemical hepatocarcinogenesis in rats and its anti-inflammatory actions.METHODS The expression of tumor marker, placental form glutathione S-transferase (GST-P), p53 and p21ras proteins were investigated by immunohistochemical techniques and ABC method. Anti-inflammatoryactivities of BR were observed by xylene and croton oil-induced mouse ear edema, carrageenin, histamineand hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet-induced mousegranuloma formation methods.RESULTS The 500 mg/kg of BR-H2O extract fractionated from BR-Methanol extract had inhibitory effecton the formation of DEN-induced GST-P-positive foci in rat liver and the expression of mutant p53 and p21fasprotein was lower than that of hepatic preneoplastic lesions. Both CH2Cl2 and H2O extract from BR haveinhibitory effect in xylene and croton oil-induced mouse ear edema. BR-H2O extract exhibited inhibitoryeffect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in ratsand cotton pellet-induced granuloma formation in mice.CONCLUSION BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in earlystage of rat chemical hepatocarcinogenesis. Both CH2Cl2 and H2O extract from BR exerted anti-inflammatory effect in rats and mice.
作者: 刊期: 2000年第03期
AIM To evaluate the role of intraoperative cholangiogram (IOC) in preventing biliary duct injury duringlaparoscopic cholecystectomy.METHODS Injury location, mechanism, time of detection, treatment outcome, and whether anintraoperative cholangiogram was performed were evaluated in 31 cases of bile duct injuries.RESULTS Cholangiograms were done in 22 cases, but they were misinterpreted in 3 of them. In 12 of 19misidentified cases, the cholangiogram was interpreted correctly, and the injury detected intraoperatively.Primary laparoscopic repair or open repair and T-tube drainage solved the problem. No long-termcomplications occurred. However, in 3 of the 19 cases the cholangiogram was misinterpreted and in 4 of the19 cases no cholangiogram was performed. Three of the seven patients required a cholangioentericanastomosis. In 2 cases the diagnosis was delayed and one of these required a two-stage procedure. Morbiditywas increased. Three cases of clim impingement of the common duct had delayed diagnoses, and two of themhad injuries. Thermal injury developed in 4 cases who had cholangiograms.CONCLUSION Routine IOC plays no role in inducing, preventing, detecting, or minimizing any of theinjuries due to clips, lacerations, or electrocautery, IOC does not prevent injuries due to ductmisidentification either. Careful interpretation of IOC would prevent injuries and avoid an open operation.
作者: 刊期: 2000年第03期