Objective To explore the protective effects of schisandrin B (Sch B) on hypoxia injury induced by cobaltous chloride (CoCl2) in human proximal renal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells were randomly assigned to four groups:control group (Con, cells were untreated), CoCl2 group (CoCl2, cells were treated with 600μmol/L CoCl2 for 24 h), Sch B pretreat group (CoCl2+Sch B, cells were pretreated with 1μmol/L and 10μmol/L Sch B for 2 h) and Sch B group (Sch B, cells were treated with 1μmol/L and 10μmol/L Sch B for 2 h). CCK-8 kit was used to detect the cell viability of four groups. Flow cytometry was used to detect the apoptotic rate of four groups. The protein expression of hypoxia-inducible factor 1α(HIF-1α) was assessed by Western blot assay. The expressions of HIF-1α and inducible nitric oxide synthase (iNOS) mRNA were determined by RT-PCR. Results Compared with the control group, after treated with 600 μmol/L CoCl2, the cell viability was decreased, and the apoptosis was increased, the expressions of HIF-1α and iNOS mRNA were up-regulated in HK-2 cells. There was no significant difference in the expression of HIF-1α mRNA between control group and CoCl2 group. Compared with the CoCl2 group, after pretreated with 1μmol/L and 10μmol/L Sch B, the cell viability was increased and the apoptosis was decreased, the expressions of HIF-1α and iNOS were down-regulated in HK-2 cells. There were no significant differences in the cell viability and apoptotic rate between control group and Sch B group. Conclusion Pretreatment with Sch B can reduce the apoptosis of HK-2 cells by inhibiting the expression of HIF-1α and iNOS mRNA, which shows protective effects on hypoxia injury.

Objective To analyze the characteristics and hepatotropism of this negative element HBVnt453-250 sequence.Methods pHBv453-250,pHBV250-453.plucHBV453-250 and plucHBV250-453 were constructed,with luciferase and enhanced green fluorecence protein(EGFP)gene as the reporter gene,respectively.After transfection of HepG2 cells with these plasmids,luciferase assays,real-time PCR and western blot assays were used to detect the gene transcription and expression level.The SV40 promoter of pGL3 control and pHBV453-250 were replaced by the cytomegalovirus early promoter,resulting in plasmids pCMVcontrol(luc)and pCMV453-250(luc).Results Compared with pHBV453-250,the mutant plasmids.with the inhibitory element inserted in difierent site or inverted orientation.exerted similar downregulation of Juciferase gene transcription and expression.Western blot analysis demonstrated the similar repression when EGFP was used as the reporter gene.By transfeeted to HepG2 cell line,the plasmid pCMV453-250(1UC)could reduce lneiferase activity(36.56%)compared with pCMLcontrol(luc).When the plasmids plueHBV453-250 and plucHBV250-453 were transfected to non-liver cell lines(A549,HeLa),luciferase gene was expressed weakly,compared with that of pGL3control(P＜0.05).Conclusion The inhibitory effect of HBVnt453-250 sequence acted in both orientation-and position-independent manners,and had no promoter selectivity and funotioned in hepatocyte-independent manner.

Objective To screen and identify proteins that interact with the hepatitis C virus core protein by means of T7-phage display system. Methods The hepatitis C virus core protein was expressed by prokaryotic expression and used as selected molecule to biopan the T7 human liver cDNA library. The selected positive clones were identified by DNA sequence and analyzed with BLAST program in GenBank. Results After BLAST in all positive clones, one protein--Smad interacting protein 1 (SIP1) was found to interact with the hepatitis C virus core protein. Conclusion T7-phage display system is a convenient, rapid and effective method for screening interacting proteins.

Objective To evaluate the real-time genotyping and quantitative PCR(RT-GQ-PCR)method by comparing it with direct sequence analysis and the multiplex-PCR method.Methods RT-GQ-PCR,direct sequence analysis and the multiplex-PCR method were used to detect HBV genotypes of 113 patient samples with HBV-DNA positive.ResultsThe detection rate of RT-GQ-PCR and direct sequence analysis was 100%,and the multiplex-PCR is 94.69%.The concordance between RT-GQ-PCR and the multiplex-PCR is perfect(Kappa value =0.915),and the consistency of RT-GQ-PCR and direct sequence analysis is pretty good(Kappa value = 0.742),specially at detecting single genotype.Twenty-eight samples with genotypes B and C dual infections were detected by RT-GQ-PCR,but only 19 samples by the multiplexPCR and 13 samples by direct sequence analysis.Conclusion The RT-GQ-PCR is convenient,rapid and accurate in HBV genotyping,especially more sensitive than direct sequence analysis and the multiplex-PCR for detecting dual genotypes.The method is applicable for large-scale epidemiological study.

Objective To study the application effect of fibrin sealant(FS) during operation for late-stage portal hypertension caused by schistosomiasis.Methods From Jun,2003 to Jun,2005,92 cases of(late-stage) portal hypertension caused by schistosomiasis treated by portal-systemic disconn-ection(PSD)(operation) were divided into two groups,namely PSD group and PSD+FS group.The early complications such as fever and exudate in splenic fossa,as well as long term complications such as recurrent bleeding,(hypertensive) gastropathy were compared in the 2 groups,and encephalopathy were compared between 64 cases of patients with FS and 28 cases of patients without FS.Results The patients undergoing PSD+FS showed decreased the exudate in splenic fossa and fever(P0 05).Conclusions FS application during operation is effective in attenuating early and long term operative complications of portal hypertension caused by(schistosomiasis),and improving outcome of surgical treatment.

Objective To analyze the expression of Galectin-3 in thyroid follicular tumors and its clinical significance in differential diagnosis between benign and malignant thyroid follicular tumor.Methods The expression of Galectin-3 in 120 cases of thyroid follicular carcinoma and 80 cases of thyroid follicular adenoma was detected by immunohistochemical technique.Results The expression rate of Galectin-3 in thyroid follicular carcinoma was significantly higher than that in follicular adenoma(84％ vs 19％,P ＜ 0.01).Strong positive expression of Galectin-3 was found in 40％ cases of thyroid follicular carcinoma and no case of thyroid follicular adenoma.Conclusions The expression of Galectin-3 can be regarded as an important parameter for differential diagnosis between benign and malignant thyroid follicular tumor.

Objective To explore the clinicopathologic characteristics and treatment of medullary thyroid carcinoma(MTC).Methods The data of 42 cases of MTC admitted to Fujian Medical University Provincial Clinical College from Jan 1995 to Feb 2010 were retrospectively analyzed.Surgery was performed in all the 42 cases.All were proved to be MTC by pathology.Results Surgery was performed in all the 42 cases.All were proved to be MTC by pathology.The lymph node metastasis rate was 76.19％ (32/42).The central lymph node metastasis rate was 61.90％ (26/42).All the 42 cases were followed up for 1 to 502 months with 102 months as the median.35 cases(83.33％)were followed up for more than 5 years and the 5-year overall survival rate was 85.71％ (30/35).Death occurred in 5 cases.Conclusions The lymph node metastasis rate of MTC is high.The treatment should be based on the principle of radical resection.Total thyroidectomy should be recommended to treat primary tumor.Lymph node dissection should be as thorough as possible.Central compartment neck dissection should be done in all cases.

OBJECTIVEBy studying the possibility of obtaining expression of human hepatitis B virus (HBV) genes and production in normal liver cells from heterologous species like normal primary duck hepatocytes (PDH), to investigate the species-specificity of HBV infection and replication.

METHODSTwo days after transfecting the complete HBV genome into PDH by electroporation (transfected group), HBsAg and HBeAg in the supernatants and lysates of PDH were measured by the IMX system. Meanwhile, replication of HBV in PDH was analyzed by Southern blotting and dot blotting procedures. PDH was electroporated as control.

RESULTSHBsAg in the lysate of transfected group was 9.10 (P/N values, positive?2.1), HBeAg was 1.0 (negative?2.1), both were negative in the supernatants of transfected group. dot blotting revealed that transfected group was strongly positive, whereas the control group was negative. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (RC), covalently closed circular (CCC) and single-stranded (SS) HBV DNA replicative intermediates in the transfected group, and there was no integrated HBV DNA in the cellular genome. Control groups were negative.

CONCLUSIONSReplication of HBV can occur in hepatocytes from nonmammalian species, which strongly supports the idea that replication of HBV has no critical species-specificity, and yet it depends on the endoenvironment of hepatocyte.

Animals ; Cells, Cultured ; DNA Replication ; physiology ; DNA, Viral ; biosynthesis ; Disease Models, Animal ; Ducks ; Electroporation ; Hepatitis B Surface Antigens ; analysis ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; physiology ; Hepatocytes ; metabolism ; virology ; Humans ; Species Specificity ; Transfection ; Virus Replication

Objective To summarize the experience of diagnosis and treatment strategy of pancreatic injury.Methods The data of 36 cases of pancreatic injurics admitted to Fujian Medical University Provincial Clinical College from 1990 to 2011 were analyzed retrospectively.Results 14 cases(39％)were diagnosed preoperatively,and the other cases were diagnosed during the laparotomy.2 cases underwent non-surgical treatment.34 cases underwent surgical treatment,among whom 23 cases underwent pancreatic debridement and drainage,6 cases underwent distal pancreatectomy(4 cases undergoing distal pancreatectomy plus splenectomy included),4 cases underwent distal Roux-Y pancreajejunostomy plus proximal pancreas closure,and 1 case underwent pancreatoduodenectomy.31 cases (86％) were cured,and 5 cases died (14％).Conclusions Most of the pancreatic injury is diagnosed through laparotomy.Surgical opcration is the main approach to treat pancreatic injury.Nonsurgical treatment is primarily limited to grade Ⅰ and Ⅱ pancreatic injury without obvious peritonitis,major pancreatic duct injury,and associated injuries.Surgical procedure should be selected based on the grading scale of pancreatic injury,associated injuries and overall conditions of the patient.

The construction of "Double First-class" has started a new journey of higher education in China. Scientific, reasonable and objective evaluation on universities is the foundation and important guarantee for the construction of first-class universities. On the basis of introducing the main university evaluation system at home and abroad, the selection of representative Chinese and world university evaluation system for comparative study has strong guiding significance for the establishment of first-class university evaluation system. Under the background of "Double First-Class" construction, based on Chinese characteristics, the author puts forward some thoughts and suggestions on perfecting the university evaluation system in China.