AIM To evaluate the expression of CD44v3 and v6 protein in colorectal carcinoma and its prognosticsignificance.METHODS One hundred and twenty-one cases of formalin-fixed paraffin-embedded colorectal carcinomaspecimens were retrospectively analyzed using EnvisionTM immunohistochemical method with the monoclonalantibody CD44v3 and v6. The median follow-up time was 67.77 months and the prognostic value of theCD44v3 and CD44v6 was assessed using univariate and multivariate survival analysis.RESULTS The positive rates of CD44v3 and v6 protein were 60.3% and 57.9%, respectively. There wassignificant correlation between CD44v3 immunoreactivity and tumor location, lymph node metastasis, distantmetastasis and Duke's stage (P< 0.05, Spearman correlation test). Significant correlation between CD44v6immunoreactivity and patients' gender, lymph node metastasis, distant metastasis, Duke's stage was alsonoticed (P < 0.05, Spearman correlation test). The 5-year survival rates were 81.25% and 60.27% inCD44v3 negative and positive cases, respectively. As CD44v6, the 5-year survival rates were 80.39% and60.00% in CD44v6 negative and positive cases, respectively; these differences between the two groups ofpatients were significant (P<0.05, Log-rank test). In multivariate analysis using the Cox regression model,CD44v3 expression emerges as an independent prognostic indicator.CONCLUSION CD44v3 and v6 might play some important roles in metastasis of colorectal carcinoma, andCD44v3 expression might be a new useful independent prognostic marker of colorectal carcinoma.

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

AIM To compare the point mutation deviations of HGV among E2, NS3 and NSSA.METHODS Seven patients with hepatic diseases from Japan and China were selected for this study. RNAwas extracted and amplified by semi-nested RT-PCR; and the PCR products were sequenced directly.RESULTS The point mutation deviations of HGV ia E2, NS3 and NS5A were 10% - 17%, 11% -23%,and 0% - 5%, in nuclcotide sequences and 4% - 12%, 0%, and 0% - 6% in amino acid sequencesrespectively.CONCLUSION The frequency of variation at the nucleotide level was in the order NS3>E2>NS5A, whileat the amino acid level the order was E2 >NS5A>NS3. The detected sequences from the N-terminus of E2may be the poorly conserved region of HGV.

作者： 刊期： 2000年第03期

AIM To compare the effects of intravenous route and peritoneal route on liver targeted uptake andexpression of plasmid delivered by glyco-poly-L-lysine (G-PLL).METHODS The plasmid pTM/MMP-1 which could be expressed in eukaryotic cells was bound to thegalactose-terminal glyco-poly-L-lysine (G-PLL), and then was transferred into Wistar rats by intravenousand intraperitoneal injection respectively. Afterwards the expression and distribution of the plasmid wereobserved at different time points by in situ hybridization and immunohistochemistry.RESULTS The plasmid could be expressed obviously in 24 hours after being transferred in vivo by bothintravenous and intraperitoneal route. One week later the expression began to decrease, and still could beovserved three weeks later. Although both the intravenous and intraperitoneal route could deliver theplasmid to liver targetly, the effect of the former was better as compared with that of the latter.CONCLUSION Intravenous route was better for liver targeted uptake and expression of G-PLL-boundplasmid than peritoneal route.

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 investigate the diagnostic significance of cytology and telomerase activity in the exfoliated cells ofcardia obtained from endoscopic brushing in the cardiac cancer.METHODS The techniques of the qualitative TRAP-silver staining and quantitative TRAP-PCR-ELISAwere employed to detect telomerase activity in the exfoliated cells of cardia obtained from endoscopicbrushing in 72 cases with cardial lesions, cytological diagnosis was made at the same time.RESULTS Telomerase activity with cardiac cancer group (1.521 ± 0. 192) was significantly higher than thatwith cardialitis group (0.065± 0.014). Positive rate of telomerase activity detected in cardiac cancer group(88.89%) was significantly higher than that with cardialitis group (11.11%), the former was significantlyhiger than cytological examination (77.78%). The diagnostic rate of cardiac cancer reached 93.33% iftelomerase activity and cytology were examined at the same time.CONCLUSION Cytology and telomerase activity in the exfoliated cardiac cells may be an effective andsensitive methods in the diagnosis of cardiac cancer. This research can be a basis for the mass screening ofcardiac cancer.

作者： 刊期： 2000年第03期

AIM To study the clonality of the esophageal carcinosarcoma by using molecular approaches.METHODS Two esophageal carcinosarcomas were included in the study. Tumor area from dysplasticlesion, squamout cell carcinoma, basaloid cell carcinoma and spindle cell elements were microdissectedseparately. Each element was analyzed with 14 microsatellite markers and direct sequenced for p53 gene andras gene mutation.RESULTS Both tumors displayed a typical histologic feature of carcinosarcoma. Both cases showed thedivergent differentiation by immunohistochemistry study. In case 1 the identical LOH at p53 and hMLH1 lociwas detected. The heterogenous LOH was detected only in carcinosarcoma at RB1 and BRCA1 loci, whilethe LOH at ACTC locus was seen only in sarcoma. The same mutation of the splice site of exon 6-intron 6displayed in the two tumor elements. In case 2, a coordinate LOH at RB locus was demonstrated in threetypes of tumor elements: sqamous carcinoma, basaloid carcinoma and spindle cell element. A heterogenousLOH was seen only in spindle cells at TAP1 locus. No mutation in exon 5-8 of p53 gene has been found incase 2. No mutation of K-ras gene was found.CONCLUSION Although the different differentiation, the two elements of esophageal carcinosarcoma mayhave a single clonality. The p53 gene mutation occurred before the two differentiation directions switched.The distinct molecular genotype can be determined through molecular biological analysis. The microsatelliteprofiling can serve as an approach to find out which genetic alteration occurs before or after thedifferentiation is determines.

作者： 刊期： 2000年第03期

AIM To study the liver-protecting and fibrosis-resisting effect of Ganxianning (GXN) and its mechanism.METHODS Model of carbon tetrachloride hepatic injury fibrosis rats was reproduced. In the experimentthere were six groups, the treatment groups with GXN's large, moderate and small dose (GXNb, GXNm andGXNs), the treatment group with colchicine, the blank model group and normal control group. The course of treatment was 30 days, then the rats were killed with their blood and liver tested.RESULTS In treatment groups, alanine aminotransferase (ALT) was lower than that in the model group(P<0.01), and albumin (Alb) higher than that in the model (P<0.01). Hydroxylproline (Hyp) and redcell membrane C3B receptor garland in GXNb's and GXNm's groups were lower and circulation complex(CIC) was slightly higher. Fibrinogen (Fb) in both colchicine and model groups was higher than that innormal group and the difference was significant (P<0.05, P<0.01). Compared with model group, acid-α-naphthyl acetate esterase (ANAE) increased in GXNb's and GXNm's groups (P<0.05, P<0.01). Underlight and electron microscopes, level of hepatic fibrosis of GXN groups was much lower than that of themodel group, P<0.01, and their difference was very significant. In GXNms group, liver cell was normal onthe whole and its chromatin was more than the model group and its nucleolus was evident.CONCLUSION GXN has rather good functions of protecting liver and resisting fibrosis, and thesefunctions are related to the increase of ANAE and C3b, decrease of CIC and Fb. and improvement of bodyimmunity function.

作者： 刊期： 2000年第03期

AIM To study the effect of sclerotic complex agents (SCA) on the gallbladder wall of hybrid rabbits, andits therapeutic effect in hepatic cysts.METHODS The SCA containing tetracycline and dexamethasone was injected into the gallbladder of rabbitsto compare its action with those of normal saline and absolute ethylalcohol on the gallbladder wall. Thetherapeutic effects of SCA and absolute ethylalcohol on hepatic cysts were observed.RESULTS Abnormal changes were not found in the tissue cells of gallbladder in normal saline group. Butin absolute ethylalcohol group, a large amount of oozing fluid and blood appeared, the absorption processwas slow, and the fibrous tissue proliferated scarcely. In SCA group, there was less oozing fluid, no blood inthe gallbladder, and the absorption was active and the fibrous tissues grow obviously. In clinical practice,SCA possesses much advantage in the treatment of hepatic cysts, by which the cysts closed promptly, theexudation reduced from early stimulation, and no relapse occurred. The cure rates at the third, sixth,twelfth and twenty-fourth month were 65.1%, 96.2%, 98.1%, and 99.1% respectively; while in thecontrol group were 10.8%, 36.0%, 67.6% and 88.3% respectively. The difference was significant(P<0.01, <0.01, <0.01, <0.05). After the observation for 24 months, no relapse occurred in the SCAgroup, but 7 (6.3%) cases relapsed in control group (P<0.05).CONCLUSION The sclerotic agents should be used in sequence, i. e., a high concentration wasadministered to reduce and destroy the epithelium of the cysts, and to promote fiber tissue adhesion and thenthe remaining drug was used to stimulate epithelium to absorb the exudation. This combined regimen wasproved to be an ideal and effective method for treating hepatic cysts clinically.

作者： 刊期： 2000年第03期

AIM To investigate the morphological and ultrastructural changes in the human gastric carcinoma cell lineBGC-823 after being treated with tachyplesin.METHODS Tachyplesin was isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus)hemocytes. BGC-823 cells and the cells treated with 2.0 μg/mL tachyplesin were examined respectively withlight microscope, scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone restorative morphological and ultrastructural changes after beingtreated with 2.0 μg/mL tachyplesin. The cells tended to be flat and spread, and their volume enlarged,nucleo-cytoplasmic ratio decreased, the shape of nucleus became relatively regular, the number and volumeof nuleous decreased, heterchromatin decreased while euchromatin increased, the number of mitochondriaincreased with their structure relatively consistent, Golgi apparatus turned to be typical, rough endoplasmicreticulum increased, polyribosome decreased, microvilli and filopodia reduced while lamellipodia increased.CONCLUSION Tachyplesin could change the malignant morphological and ultrastructural characteristics ofhuman gastric carcinoma cells effectively and had certain effects on inducing differentiation of human gastriccarcinoma cells.

作者： 刊期： 2000年第03期

AIM To study the effect and alteration of hepatic blood flow as well as ultrastructure of hepatic tissue inchronic cholestatic hepatitis after hypebaric oxygenation (HBO).METHODS Using the hepatic rheometer and Doppler B-mode ultrasound equipment, the contractive waveof hepatic blood flow and blood flow of portal vein were tested; the biochemistry, immunohistochemistryand ultrastructure of hepatic tissue were determined and served as the evaluating indexes.RESULTS After the HBO treatment, the contractive wave of hepatic blood-flow in 76% patients and bloodof right portal vein in 70% patients were increased, the improvement of serum ALT and BILI was 88.9%and 93.3% respectively. In addition, the swelling mitochondria, cholestasis of hepatic cells and capillariesreduced obviously; Kupffer's cells decreased. There was significant difference (t=2.85, P<0.05) beforeand after HBO treatment.CONCLUSION It is suggested that the HBO could increase the blood flow of portal vein and arteries,improve the hepatic function, cholestatsis and inflammation.

作者： 刊期： 2000年第03期

AIM To explore the anti-tumor effect of indomethacin (IN) on human colon adenocarcinoma cells anddetermine the influence of indomethacin on cancer cell proliferation and apoptosis and elucidate the anti-tumor mechanism of Indomethacin.METHODS Human colon adenocarcinoma HCT116 cell line were cultured separately in vitro. Indomethacin(final concentration 100 μm - 800 gm) was administered alone or altogether with 5-Fu (50 μm). Agarose gelelectrophoresis, MTT, and Flow cytometry were used to study cell proliferation and apoptosis in humancolon carcinoma cell RT-PCR, western blot were used to detect the expression level of Bcl-2, bax gene andcdk4 protein expression in HCT116 cell lines after treated with IN for 24 hours.RESULTS Indomethacin can inhibit significantly the proliferation of HCT116 cell, change the morphology,and cause the cells to accumulate in the G0/Gl phase of the cell cycle, and induce apoptosis. The apoptosis oftumor cells was confirmed by DNA ladder formation on gel electrophoresis and sub-Gl peak on flowcytometry. These responses were time-and concentration-dependent. A synergic effect of inhibiting cancercell proliferation was observed when combined with Indomethacin and 5-Fu. RT-PCR results showed that INdown-regulated Bcl-2 mRNA expression, and did not change Bax mRNA expression. Western blot resultsconfirmed that IN inhibited Bcl-2 protein expression. No influence was found in the translation of Baxprotein. In inhibited cdk4 protein expression.CONCLUSION Our study results indicate that IN induce apoptosis of HCT116 cell by down-regulating Bcl-2expression and inhibiting cdk4 protein expression partially. This explains the mechanisms of antitumoractivity of the Indomethacin.

作者： 刊期： 2000年第03期

AIM To explore gallbladder circadian rhythm (GCR) and the cholagogic action of Yin Chen Hao Ta(YCHT).METHODS Ultrasonography of rabbit gallbladder at 17:00 to 19:00, 23:00 to 01:00, 05:00 to 07:00 m11:00 to 13:00 showed its maximum area (MA), dilatation rate (GDR) and systole rate. Its movemecircadian rhythm (MCR) and the effect of YCHT on gallbladder and the relationship with time weobserved and evaluated.RESULTS The results showed that gallbladder area changed with time, which varied from 11:00 to 13:1>23:00 to 01:00 and 05:00 to 07:00> 17:00 to 19:00 (P<0.01, F=9.13 vs. control by analysisvariance). The cholagogic action at 23:00 to 01:00 and 17:00 to 19:00 was better than that at 11:00 to 13:and 05:00 to 07:00.CONCLUSION The gallbladder movement action (GMA) showed markedly circadian rhythm. Tcholagogic action of YCHT at 23:00 to 01:00 and 17:00 to 19:00 is better.

作者： 刊期： 2000年第03期

AIM To explore the therapeutic potential of antisense oligodeoxynucleotides on hepatcellular cacinoma(HCC).METHODS Four antisense phosphorothioated oligodeoxynucleotides (asON), complementary to differentsites of HBV, were synthesized and assayed for their anti-HBV activity in HepG22. 2.15 cells with ELISA.The most effective asON was chosen for the following study: FACSCAN, TRAP and immuno-staining wereused respectively for checking apoptosis, telomerase activity and expression of oncogene p21ras and p62C-myc inHepG2.2.15 cells after treated by asON.RESULTS The oligomer directed against the initiator of pre-S2 was the most effective one with aninhibitory rate of 66% on HBsAg and 91% on HBeAg (P<0.02). Two inhibitory peaks (bimodal)appeared. Telomerase activity as well as the expression of p21fas and p62C-myc decreased drastically 3 days afterasON-HBpreS2 treatment. Meanwhile, apoptosis appeared in the experiments.CONCLUSION The inhibitory effects of as-preS2 on the HBV gene expression and the reversion of somemalignant behaviour in HepG2.2.15 cells were the significant, effective therapy against HBV infection andhepatocellular carcinoma.

作者： 刊期： 2000年第03期

作者： 刊期： 2000年第03期

AIM To study the physico-chemical properties of biliary calculus and the relationship between the calculusformation and the phase change of liquid crystal, providing the best evidence for the biliary calculusprevention and treatment.METHODS The cholesterol contents in thirty one cases of biliary calculus in Kunming were determined bydouble-wave-length TLC scanning with high efficiency silica gel films.RESULTS Under magnifiers, the granular biliary calculus from 31 patients were classified according totheir section structures and colours, as cholesterol cholelith, 25 cases; bilirubin cholelith, 4 cases andcompound cholelith, 2 cases. By TLC scanning, it was found that the content of cholesterol in human biliarycalculus was 71%- 100%, about 80% cholesterol bilestones whose cholesterol content was more than 90%being pure cholesterol bilestones.CONCLUSION Cholesterol bilestone is the main human biliary calculus in Kunming, which was inaccordance with X-ray analysis. Compared with the related reports, it is proved that the proportion ofcholesterol bilestones to biliary calculus is increasing because of the improved life standard and the decreaseof bilirubin bilestones resulted from bile duct ascariasis or bacteria infection in China since 90s, and that theincrease of cholesterol in-take leads to the increase of cholesterol metabolism disorder

作者： 刊期： 2000年第03期

AIM To find out if there is any difference in human primary liver carcinogenesis between Han and minorityethnic patients in Xinjiang.METHODS Expression of p53, c-erbB-2, H-rasp21 protein and proliferating cell nuclear antigen (PCNA)in tumor tissues of 50 patients (Hah 38, minorities 12) with primary hepatic carcinoma (HCC) was detectedby immunohistochemistry (LSAB).RESULTS The positive frequency of p53, c-erbB-2, H-rasp21 and PCNA expression was 46.0% (23/50,70.0% (35/50), 68.0% (34/50) and 82.0% (41/50) in tumor tissues; 4.0% (2/50), 22.0% (11/50),64.0% (32/50) and 52.0% (26/50) in peritumor respectively with a significant difference, except for H-rasp21 (P<0.05) between tumor and non-tumor tissues. Combined the three oncogenes alteration, 26%(13/50) tumor tissues had positive immunoreactivity, but peritumor and normal liver were negative. Thepositive p53, c-erbB-2, H-rasp21 protein expression was 39.5 % ( 15 / 38), 60.5 % (23 / 38) and 39.5 % ( 15 /38) in tumors of Han patients; 66.7% (8/12), 100% (12/12) and 75.0% (9/12) in minority patientsrespectively. A statistical difference between Han and minority cancer samples was observed (P< 0.05).CONCLUSION Overexpression of p53, c-erbB-2 and H-rasp21 in human primary liver carcinoma is animportant biomarker of genetic alteration. The different frequency of these oncogenetic changes may reflectsome environmental factors or/and ethnic hereditary affecting the liver carcinogenesis. The special life styleof Han, Uygur, Kazak and Mongolia nationalities in Xinjiang may also involve the etiopathogenesis of thisdisease.

作者： 刊期： 2000年第03期

AIM To establish a model system for studying gastric carcinogenesis of MNNG, a gastric cancer relatedcarcinogen.METHODS Cell culture transformation, PCR-restriction fragment length polymorphism (PCR-RFLP),DNA blotting and immunochemical techniques and analysis of LDH isozyme and chromosome wereperformed.RESULTS GES-1 cells surviving by MNNG treatment were named MC (2 × 105M for 24 hours) and MC-B(2 × 10-7M for 7 days). The two cell lines treated by MNNG showed more malignant than maternal cell GES-1 with the evidences of more chromosome aberrations, abnormal morphology and eytoskeleton and alsogained the ability of colony formation on soft agar. C-Ha-ras gene point mutation in the 12th codon and LDHisoenzyme abnormal express were found in MC-B cells. In addition, C-met gene rearrangement was revealedby Southern blot analysis in MC-B and MC.CONCLUSION This gastric epithelial cell system is an important model system for further study of stomachcancer, MNNG had a selective effect on the cytoskeleton mierofilament in human gastric epithelial cells andintimately associated with the activation of certain oncogenes and some protein.

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

Advances in molecular biology made possible the discovery of the virus that causes hepatitis C. However,little is known about the fundamental aspects of hepatitis C virus (HCV) replication, primarily because arobust cell culture has not been established. As a result, the currently available drugs for the treatment ofhepatitis C are not specifically directed against HCV. Based on what is known about the molecular biology ofHCV, however, drugs can now be developed against specific viral and cellular targets. The next generationof drugs for the treatment of hepatitis C will likely be directed against non-structural HCV proteins withknown enzymatic activities, such as the proteases, RNA helicase and RNA polymerase. Others agentstargeted against the viral RNA, core protein that assembles into the virion capsid and putative cellular“receptors” that bind HCV envelope proteins are also being developed. These drugs should have fewer sideeffects than those currently available and be much more effective for the treatment of chronic hepatitis C.

作者： 刊期： 2000年第03期