Objective: To construct and characterize GFP-mfgl 2 fusion protein expression plasmid (pEGFP-mfgl 2) and provide a direct and simplified methodology for primary assessment of the effect of mfgl 2 siRNA on the mfgl 2 gene expression. Methods: mfgl 2 cDNA was amplified from the mfgl 2 cDNA library pBluescript-m166 (pm166) of mouse genomic P1 plasmid and recloned into pEGFP-N2 upstream of GFP gene. The pEGFP-mfgl 2 was analyzed by restriction endonucleases BamH I and Hind III to ensure the orientation and the sequence. This fusion plasmid was then transfected into CHO cells and the fusion protein expression was observed by fluorescent microscope. Rusults: A 1.3 kb long cDNA was obtained. Restriction endonucleases and sequencing assays showed the correct orientation and sequence. After 24-48 hours transfection in CHO cells, the expression of pEGFP-mfgl 2 can be visualized through fluorescent microscope. Conclusion: pEGFP mfgl 2 has been constructed successfully. The recombinant vector can express GFP-mfgl 2 fusion protein. It provides a direct and simplified methodology for primary assessment of the effect of mfgl 2 siRNA on the mfgl 2 gene expression.

Objective To evaluate the prophylactic effect of pancreatic duct stent (PPDS),NonSteroid Anti-Inflammtory Drugs (NSAIDs),and joint PPDS and NSAIDs on post endoscopic rectrograde cholangiopancreatography(ERCP) Pancreatitis(PEP) in choledocholithiasis patients.Methods A total of 200 choledocholithiasis patients were randomly divided into 4 groups,prophylactic pancreatic duct stent(PPDS) group (A),NSAIDs group (B),joint PPDS-NSAIDs group (C) and routine ERCP without prevention for PEP(group D).VAS score,levels of amylase in serum and CRP were measured before and 4 h,24 h,48 h after ERCP.Incidences of hyperamylasemia and PEP were observed.Results (1) Incidences of hyperamylasemia 48 h after ERCP were 6％ (3/50),6％ (3/50) and 4％ (2/50) in group A,group B and group C respectively,which were significantly lower than that of group D (11/55) (P ＜ 0.05).(2) Incidences of PEP 48 h after ERCP were both 2％ (1/50) in group A and group C,which were lower than that in group D (10％,5/50,P ＜ 0.05).Group B (4％,2/50) was lower than that of group D but there was no statistical significance(P ＞0.05).(3) VAS scores of all groups at 4 h,24 h and 48 h after the operation were significantly higher than before (P ＜ 0.05).Group B score was significantly lower than that of group D (P ＜ 0.05).Scores of group A and C at 4 h were lower than those of group D (P ＜ 0.05),and those at 24 h and 48 h were also lower but with no statistical significance (P ＞ 0.05).(4) Serum CRP levels at 4 h,24 h and 48 h were significantly higher than those before in each group.Serum CRP levels of group B and C were significantly lower than that of group D at 4 h,24 h and 48 h.Serum CRP level of group A was significantly lower than group D at 4 h,24 h.CRP level at 48 h of group A was lower than that of group D,but there was no statistical significance (P ＞ 0.05).Conclusion Both prophylactic pancreatic duct stent and NSAIDs (Parecoxib Sodium) can reduce incidence of hyperamylasemia after ERCP common bile duct lithotomy.Single or joint use of prophylactic pancreatic duct stent can prevent PEP.Furthermore,prophylactic pancreatic duct stent and NSAIDs (Parecoxib Sodium) can reduce pain and inflammation after ERCP common bile duct lithotomy.NSAIDs only (Parecoxib Sodium) is more effective than prophylactic pancreatic duct stent only and joint use of both.

The primary causative factors of liver failure include direct damage and immune -mediated liver injury.Increasing evidence sug-gests that immune -mediated injury plays a pivotal role in the pathogenesis of liver failure.The new concepts concerning the mechanisms of immune -mediated liver injury in liver failure are reviewed with relevant basic and clinical studies in both humans and animals.The innate and adaptive immunity,particularly the interaction of various immune cells and molecules,as well as apoptosis -related molecules,are dis-cussed in detail.

Objective To study on the method of applying laparoseopy for the treatment of choledo cholithiasis after biliary tract operation, and to evaluate its therapeutic efficacy. Methods Thirty-seven cases of common bile duct stones after operation from October 1999 to December 2003, received laparoscopic common bile duct exploration (LCBDE) were retrospectively analyzed. Results Satisfactory results were accepted in 35 cases and the rest two cases converted to laparotomy. Thirty cases had got primary healing by the help of employing endoscopic nasobiliary drainage (ENBD) , 7 cases received T-tube drainage. There was neither residual stones left nor serious after-effect including biliary fistula and hemobilia happened. Mean operative time was ( 168 ?30. 2) min. Mean postoperative stay in hospital was (11.5?1.3) d. Recurrences of calculi and bile duct stricture never happened within 6-48 m follow-up study. Conclusion The laparoscopic procedure is technically feasible and safe alternative, however, the technique in lyses of adhesions, dissection and suture of choledochus are quite difficult, therefore it is recommended to be performed in experienced hands expected to minimize the occurrence of complications, and firmly mastered the indications of conversion to laparotomy as far as possible.

Objective To evaluate the efficacy of laparoscopic and open operation combined with endoscopic nasobiliary drainage(ENBD) in the management of choledocholith. Methods 44 cases treated by laparoscopic common bile duct exploration (LCBDE) and 34 ones by open common bile duct exploration(OCBDE) combined with ENBD and primary suture of common bile duct were retrospectively analyzed from January 1997 to July 2001. Results All cases were operated on successfully. The clearance rate of stones was 100%(78/78). No postoperative complications such as bile leakage, hemobilia, cholangitis, etc occurred. Postoperative hospital stay was (6 2?1 3)days in group LCBDE and (7 4?2 1)days in group OCBDE. 57 cases had been followed up for (1～4 5)years with an average of 2 6 years, and ultrasoundgraphy showed no bile duct stricture and recurrent stones. Conclusions LCBDE or OCBDE combined with ENBD and primary suture of common bile duct is safe and reliable.

Key trial activities include: development of the trial protocol;development of standard operating procedures;development of support systems and tools;generation and approval of trial information documents;selection of trial sites and the selection of properly qualified,trained,and experienced investigators and study personnel;ethics committee review and approval of the protocol;review and approval by applicable regulatory authorities;enrollment of subjects into the study: recruitment,eligibility,and informed consent;the investigational product(s): quality,handling,and accounting;trial data acquisition: conducting the trial;trial data acquisition: conducting the trial; safety management and reporting;monitoring the trial;managing trial data;quality assurance of the trial performance and data;reporting the trial.

Insulin receptor substrate(IRS)-1 and IRS-2 expression levels of liver tissues and skeletal muscle in intrauterine growth retarded(IUGR)rats were investigated by RT-PCR and immunohistochemistry.An IUGR animal model was established by maternal nutrition restriction during pregnancy.IRS-2 expression level of liver tissue and IRS-1 expression level of skeletal muscle in IUGR rats at 0 and 3 weeks old were significantly lower than those in normal rats at the same age respectively,and insulin resistance was induced in IUGR,and these findings might be the molecular mechanisms susceptible to metabolic syndrome in IUGR rats.

Objective To enhance the level of the diagnosis and treatment of relapsing polychondritis(RP).Methods Clinical data of 17 patients of RP were studied retrospectively. Results Of the patients,ear was involved in 16 cases, nose in 10 cases, laryngotracheobronchial region in 9 cases, eyes in 8 cases, joints in 4 cases, cardiovascular system in 1 case.Of 17 cases-1 died from respiratory complications. Fifteen patients had recurrence over twice,1 patient had sixth recurrence. Conclusions RP is rare autoimmune system disease, early clinical manifestation are nontypical, misdiagnosis and missed diagnosis. Corticosteroids, immunosuppressive agents, dapsone and surgical operation are used in these patients to control symptoms.

According to bias in codon choice of Pichia pastoris, The phytase phyA gene from Aspergillus niger N25 was mutated without changing its amino acid sequence. The expression plasmid pPIC9k-phyAm was constructed and transformed into GS115 strain. Positive clones,of which the chromosomes were integrated with phyA gene,were identified by the phenotype and PCR. SDS-PAGE analysis suggust that the size of enzyme protein of the expression product was about 70.15kDa.Southern blotting analysis to the yeast transformants showed that phyA gene was intergrated into the chromosome genome. The phytase activity of PP-NP m-4-4 with codons optimized reached 136 000U/ml in malt wort culture medium after being induced with 36h, which was the 2.8 times of the original strain PP-NPm-8.

Objective:To clarify the contribution of endogenous bFGF, bFGF mRNA and FGFR1 to the abnormal growth and phenotypic transformation of neoplastic tumors cells.Methods:The antisense oligonucleotide primers was used to evaluate the influence of endogenous bFGF on growth of human glioma malignant cell lines SWO-38 in vitro. MTT was used to examine the variety of cells growth treated with bFGF antisense oligonucleotide primers. The methods of ELISA, in situ hybridization, immuno-hischemistry and image analysis were used to detect the expression level of bFGF, bFGF mRNA and FGFR1. The colony formation of cells in soft agar was used to assess the cloning efficiency of the cells after exposed to bFGF antisense oligo-nucleotide primers.Results:The cells multiplication, expression of bFGF mRNA and FGFR1 was inhibited by bFGF antisense oligonucleotide primers,and the cells multiplication was dose-dependent. Treated with antisense oligo-nucleotide primers, the expression of FGFR1 and secretion of bFGF were distinctly reduced, and the inhibition efficiency of cells multiplication of WSO-38 was 48% and the inhibition efficiency of colonies of SWO-38 in soft agar was 35%. The inhibition of cells multiplication can be reversed completely by external bFGF, and the reverse efficiency was 8%.Conclusion:The synthesis of bFGF mRNA and expression of bFGF can be specifically inhibited by antisense oligonucleotide, but the inhibition can be cleared up with the addition of external bFGF. The study suggested that the bFGFantisense oligonucleotide could have good effect in inhibiting of tumor under special condition.