In this study, alpha-fetoprotein variant (AFP variant) in serum and tumor tissue from patients with hepatocellular carcinoma (HCC) was determined by lectin crossed-immuno-affino-clectrophoresis (LCIAE). The cirrhosis and liver diseases without tumor were taken as control.The results indicate that two kinds of AFP variants were found by LCIAE method and they were different in the HCC patients from different areas,The ratios of two kinds of AFP variants in serum and tumor tissue were near, indicating there are tumor cells in liver, which can secrete two kinds of AFP variants.AFP-N-L content is closely related with tumor size, tumor cell differentiation, histological behaviors, tumor capsulc, tumor thrombus, etc.Tumor cells with high malignance secrete mainly LCA-R-AFP.

It has been concerned by the people for a long time to unfold the interruption of hepatic blood flow in security. The present work was undertaken for the clinical practice. 59 heathy adult rabbits were used in this study. These animals were divided into three groups; (1) ligating the right branch of the hepatic artery, (2) ligating tne right branch of the portal vein, and (3) ligating both the right branch of the hepatic artery and portal vein. The hepatic weight, histology, ultrastructure and serum enzymes were observed at regular intervals after the operation. Vaso-graphys were also performed in the selected animals to investigate reconstruction of collateral vessels.The experiment results;1. During the earlier stage after the ligation of the right hepatic artery, in-farct areas of various extent were occurred in the lobe from which the arterial blood supply had been excluded. Finally, they can be classified into two types, the restored type (55.6%) and the infarction type (44.4%). After the ligation of the right portal vein, the ligated lobes became atrophic, and nonligated lobes became compensatory hepertrophic, and by a balance of the two processes the original weight of the liver was maintained. After the ligation of both the right branch of the hepatic artery and portal vein, the infarction was total in lobes supplied by the occluded vessels in 83 per cent of animals. There were partial normal liver tissues remained only in three rabbits.2.On the third postoperative day, the serum GPT activity rose much higher in the animals of 3 rd group than that in the 1 st or 2 nd group.3.The collateral circulation was reconstructed at the 25 th day after ligating the right hepatic artery, while there was no collateral circulation reconstructed after ligating the right portal vein.4.The rabbits could tolerate about 20 per cent of acute infarction of liver tissues.The authors are conveined that;1.There were some dangerous after the ligation of the hepatic artery. The extent of hepatic infarction may be reduced after interrupting the hepatic arterial flow if we could take some methods to alter the stagnation of portal blood flow and to enhance the hepatic cell against hypoxymia.2.The ligation of the branch of the portal vein may be practiced in the clinic. There was no bad result to the organism.3.The ligation of both the branch of the hepatic artery and portal vein should not be performed, and this may lead to severe consequence.4.In the early period after operation dynamic observating the serum GPT activity may be used to evaluate the degree of the liver damage.

38 percutaneous transhepatic cholangio-drainages (PTCD) were performed in 35 patients, and two failed.The successful rate was 94.7%. X-PTCD was done in 18 cas s, and US-PTCD in 20 cases, their successful rates were 100% and 90% respectively. Prc-operativelv, PTCD was applied to 22 patients with biliary obstruction.The mean period of drainage was 15 days.200-500 ml of bile wero drained cvery day. PTCD was applied to 11 patients with malignent cancer. Fover appeared in 5 cas s, block of the catheter in 2 cases, dislocation of the catheter in 5 cases, and biliary peritonitis in one case.It seems that PTCD may decrease the operative mortality and prolong the survival time of the patients with obstructive jaundice.The procedure of X-PTCD can be easily performed and adopted by a beginner.US-PTCD can be performed with an intrahepatic bile duct whch is wider than 8 mm in diameter, cven without exposure to x-ray.

Microheterogeneity of alpha-fetoprotein (AFP) in the sera of 78 patients with hepatocellular cancer (HCC) and 40 patients with benign liver diseases (BLD) was studied by lectin affino-immunoelectrophoresis autoradiography. Serum AFP concentre tions varied 124-56000ng/ml in patients with HCC and 31-980 ng/ml in patient with BLD.By means of either LCA or Con-A, AFP was divided into two variants, which were coded AFP-R-L (AFP retarded by LCA) and AFP-N-L (AFP not retarded by LCA), AFP-R-C (AFP retarded by Con-A) and AFP-N-C (AFP not retarded by Con-A), respectivily. AFP-N-L percentage lower than 75% was observed in 87.2% of patients with HCC and none with BLD, AFP-R-C percentage lower than 100% in 89.7% of cases with HCC and 17.5% of patients with BLD. According to the diagnostic standard of AFP level greater than 400ng/ml for primary liver cancer, the positive rate of patients with HCC was 78.2%, and that of patients with BLD was 20.0%. Determination of microheterogeneity of AFP ran raise the positive rate of patients with HCC to 97.4% and reduce that of patients with BLD to O (by mean of LCA). Therefore, determining the concentrations of AFP variants can increase the clinic value of AFP as a marker of primary liver cancer.

Objective To study the role of Caspase-1 activated cytokines in acute lung injury (ALI) in experimental severe acute pancreatitis (SAP). Methods A rat model of SAP was induced by retrograde infusion of 5% sodium taurocholate into the biliary pancreatic duct. Thirty-two SD rats were randomly divided into four groups: healthy control (HC), SAP 6h, SAP 12h and SAP 18h groups. Serum IL-1? level was measured by ELISA. Intrapulmonary expressions of Caspase-1, IL-1? and IL-18 mRNA were detected by semi-quantitative RT-PCR. Results The serum IL-1? levels were significantly increased after the induction of pancreatitis (P

Objective To explore the potential role of the expression of cytokine-induced neutrophil chemoattractant (CINC)gene in the pathogenesis of acute lung injury(ALI)in early severe acute pancreatitis(SAP). Methods SAP was induced by retrograde injection of 5% sodium taurocholate into the bili-pancreatic duct. Wet to weight ratio and myeloperoxidase(MPO) content of lungs were measured. Pathological changes in the pancreas and lungs were observed. Intrapulmonary expression of CINC gene was assessed by reverse transcription polymerase chain reaction(RT-PCR). Results Compared with that in controls,intrapulmonary expression of CINC gene?wet to weight ratio,and MPO of lungs were increased greatly in twelve hours. The CINC mRNA expression in lungs was significantly correlated with the severity of lung injury,wet to weight ratio,and MPO of lungs. Conclusion Overpexpression of CINC mRNA in the lungs might play an important role in the pathogenesis of ALI in early SAP.

Objective To construct a recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori ( H. pylori ) hpaA gene and to detect its immunogenicity. Methods hpaA gene fragment was amplified by polymerase chain reaction (PCR) from the genomic DNA of the standard H. pylori strain and cloned into pUCmT vector. DNA sequence of the amplified hpaA gene was assayed,then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions. The recombinant plasmid was used to transform Escherichia coli DH5?,and the positive clones were screened with PCR reaction and restriction enzyme digestion. Then,the recombinant pIRES-hpaA was transformed to LB5000,then the recombinant plasmid was transformed to SL7207,and the recombinant strain was passaged repeatedly. Finally,recombinant pIRES-hpaA was transfected to COS-7 cells with Lipofectamine TM 2000. The immunogenicity of expressed hpaA protein was determined with SDS-PAGE Western blot. Results The hpaA gene fragment of the 750 base pair was amplified from the genomic DNA,and it was consistent with the sequence of the H. pylori hpaA by sequence analysis. It was confirmed by PCR and restriction enzyme digestion that Helicobacter pylori hpaA gene was inserted into the eukaryotic expression vector pIRES and a stable recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori hpaA gene was successfully constructed,and specific strip of hpaA expressed by pIRES-hpaA could be detected with Western blotting. Conclusion The recombinant attenuated Salmonella typhimurium nucleic acid vaccine strain expressing HpaA protein with immunogenicity was constructed,it might be helpful for further investigation concerning the immune action of the nucleic acid vaccine in vivo .

Objective To clone human canstatin gene, construct its prokaryotic expression vector, express and purify its recombinant protein. Methods The total RNA was extracted from human placenta tissues. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid pET-22b(+) and then transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG). The expression of protein was analyzed through SDS-PAGE. Cells after induced 3 hours by IPTG were harvested, sonicated briefly and the proteins were purified through affinity chromatography. Results (1)The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis and the concentration was 1.8 g/L. (2)The target sequences were specifically amplified through RT-PCR. (3)The purified RT-PCR product was cloned into pUCm-T vector and then sequenced, demonstrating to be the same as that of canstatin gene in GenBank. (4) the expression vector pET-22b(+) was constructed and verified by the method of BamH Ⅰ and Hind Ⅲ digestion. (5) After IPTG induction, there was a new protein band about Mr 24kD on SDS-PAGE. The percent expressed product over total bacterial proteins after 1, 2, 3, and 4 hours of induction was 18.2%, 18.8%, 23.0% and 23.4%, respectively, estimated by densitometry. (6)After affinity chromatography, SDS-PAGE showed only one clear band existed in 125 mmol/L or 250 mmol/L imidizone elution. Conclusion Human canstatin gene has been successfully cloned and its prokaryotic expression vector has also been successfully constructed. Further more, purified recombinant proteins are obtained through affinity chromatography, laying foundation for further study of its clinical application.

Objective To study clinical characteristics and prognosis of patients with liver injury of acute pancreatitis.Methods 290 cases of acute pancreatitis admitted between January 2001 to October 2003 were reviewed and analyzed retrospectively.Results Through comparing different types of AP with liver injury and C-reactive protein (CRP) changes,it was found that the more severe AP was,the more significant liver injury was;and liver injury of severe AP had some connection with CRP(.P.

Objective To explore the potential role of cytokine-induced neutrophil chemoattractant (CINC) and monocyte chemoattractant protein-1/JE (MCP-1/JE) in the pathogenesis of early acute pancreatitis(AP). Methods Acute edematous pancreatitis(AEP) and acute necrotizing pancreatitis(ANP) were induced by retrograde infusion of 0.5% or 5% sodium taurocholate into the biliary pancreatic duct. The activity of serum amylase and the level of serum calcium were studied, wet to weight ratio,the activity of myeloperoxidase (MPO) and pathological changes of pancreas were observed. The expression of gene and protein of CINC and MCP-1/JE in pancreatic constitution were detected. Results Compared with that in controls,pancreatic acinar cells were found to express CINC and MCP-1/JE protein in certain intensity. With the induction and the progress of AP,the expression of the activity of MPO,CINC and MCP-1/JE gene in pancreatic parenchyma were all significantly increased ( P