Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.

Objective To investigate the role of MHC class Ⅱ transactivator(CⅡTA) in upregulating the expression of HLA class Ⅱ antigen in HepG2 cells.Methods Eukaryotic expression vector EBS-NPL-CⅡTA containing CⅡTA cDNA or the empty vector EBS-NPL was transfected into HepG2 cells respectively.CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were respectively detecued by RT-PCR,indirect cell immunofluorescence technique and flow cytometry in original HepG2 cells,HepG2 cells transfected with EBS-NPL-CⅡTA or the empty vector EBS-NPL.Results The expression of CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were not observed in original HepG2 cells and HepG2 cells transfected with empty vector,but in the HepG2 cells transfected with EBS-NPL-CⅡTA.Conclusion CⅡTA is a switching factor of mastering the expression of HLA class Ⅱ antigen in HepG2 cells.The lack of CⅡTA expression in HepG2 cells contributes to no expression of HLAⅡ antigen.

Objective To study the effects of IFN ?,TNF ?,IL 1 and dexamethasone on the expression of intercellular adhesion molecule 1(ICAM 1) of primary cultured human hepatocyte. Methods ICAM 1 expression of primary cultured human hepatocyte was induced in vitro by IFN ?,TNF ? and IL 1, measuring by cellular enzyme linked immunosorbent assay(ELISA). The effect of dexamethasone on human hepatocyte ICAM 1 expression was observed by adding dexamethasone into the cultured human hepatocytes prior to cytokine stimulation. Results The levels of ICAM 1 expression were significantly lower in unstimulated human hepatocytes than in human hepatocytes induced by IFN ?,TNF ? and IL 1. There were relations among the levels of ICAM 1 expression and cytokine concentration as well as induced period of time. Dexamethasone could partly inhibit the ICAM 1 expression on human hepatocytes induced by TNF ?,IL 1 and IFN ?.The inhibition rates were 40.3%?5.9%,38.1%?4.8%, and 37.6%?6.7%, respectively.Conclusions Enhanced ICAM 1 expression of primary cultured human hepatocyte may be induced in vitro by IFN ?,TNF ? and IL 1, which may be partly inhibited by dexamethasone.

0.05). Conclusion The polymorphism of G-944C in CⅡTA gene promoter Ⅳ was associated with the susceptivity of chronic HBV infection, but was not associated with severity of diseases. The individuals with chronic HBV infection of CC genotype are of less possibility to develop chronic liver disease than those of other genotypes.

Objective To study the mechanism of activity change of gastric alcohol dehydrogenases(ADH).We investigated ADH before and after ingestion of alcohol on rats.Methods Thirty-nine rats were divided into two groups at random:model ones(24 cases) and control ones(15 cases).Model group of rats were infused into stomaches by alcohol,and control ones by distill water.Structures of cell and sub-cell in gastric mucous membrane were observed by enzyme electron microscope and hematoxylin eosin(HE) stain.Results It was shown that there were dropsical degeneration of chief cells and parietal cells of stomaches by HE stain,and the mitochondria of parietal cells and the rough-surfaced endoplasmic reticulum(rER) of chief cells were destroyed by enzyme electron microscope in model groups after two months of ethanol perfusion.There were not change in control group.Conclusions The mechanism of activity alterations of gastric ADH may result from dropsical degeneration of chief cells and parietal cells of stomach,which result in degeneration,solution and breakage of construction of rER in the chief cells and mitochondria in parietal cells.

Objective To investigate the relation between the prognosis and complication of patients with severe viral hepatitis (SVH) Methods The incidences of different complications in 573 patients with SVH were analyzed The survival rates of SVH patients with different complications were compared Results Complications were diagnosed in 77 8% SVH patients The incidences of hepatic encephalopathy, infection, brain edema, gastrointestinal hemorrhage, and hepatorenal syndrome were 50 4%, 34 6%, 30 2%, 22 2%, and 21 8%, respectively The survival rate of SVH patients without complications was 78 7%, significantly higher than that of patients with complications (13 7%, P

Objective To investigate the relationship between the non-homonymy single nucleotide polymorphism(SNP)of C19170G,C30799G in the coding area of class Ⅱ transaetivator(CII TA)and the different clinical phenotypes of chronic hepatitis B virus(HBV)infection.Methods Six hundred and twenty-seven patients with chronic HBV infection and 101 healthy blood donors were enrolled in this study.Genotyping of C19170G,C30799G in C Ⅱ TA gene coding region were done by sequence-specific primer polymerase chain reaction(PCR-SSP).Hardy-Weinberg balance of the genotypes was analyzed using chi-square test.Differences between two sets were tested by contingency table chi-square test and unconditional Logistic regression was performed. Results The frequencies of G allele and GG+GC genetypes at 19170 site were significantly higher in patients with liver cirrhosis than those with non-progressive liver diseases(X2=7.128,P=0.008;X2=6.404,P=0.011,respectively).There were significantly differences of the allele frequencies between patients with liver cirrhosis and non-progressive liver diseases(OR:0.742,95%CI:0.552～0.998,P=0.048),and the main differences were observed in G dominant model(OR:0.581,95% CI:0.353～0.954,P=0.032).The frequencies of C allele and CC genotype at 30799 site were significantly higher in patients with hepatocellular carcinoma than those in patients with liver cirrhosis(X2=4.861,P=0.027;X2=4.993,P=0.025).There were significant differences of the genotype frequencies at 30799 site between patients with liver cirrhosis and hepatocellular carcinoma(OR:0.557,95% CI:0.334～0.930,P=0.025),and the differences were mainly observed in C recessive model(OR:0.491,95% CI:0.269～0.898,P=0.021).Conclusions The polymorphisms at 19170 site are associated with liver cirrhosis in chronic HBV infection,and the G allele carriers are prone to progress into liver cirrhosis.The polymorphisms at 30799 site are associated with hepatocellular carcinoma in chronic HBV infection,and CC genotype carriers are prone to progress into hepatocellular carcinoma.

OBJECTIVE:To study the dynamic changes of TNF-?/IL-10in model rats with acute necrotizing pancreatitis (ANP)and to study the intervention outcome of salvia miltiorrhiza.METHODS:A total of96rats were randomly divided into three groups(each with32rats):pancreatitis(P)group,salvia miltiorrhiza treatment(T)group and control group(C),the blood samples of rats in each group were taken to determine levels of TNF-?and IL-10and the ratio changes of TNF-?/IL-10.Meanwhile,pancreas tissue sample was collected for pathological scoring.RESULTS:Group P had significantly higher serum levels of TNF-?and IL-10than did group C(P

Objective To study the effects of elodronate-liposome on inducing apoptosis of alve-olar macrophages from rats with acute neetotizing pancreatitis (ANP).Methods The AMs of eight rats with ANP were isolated, purified then incubated from broehoalveolar lavage by the differing rates of attachment of the various cell types in a forty-well cell culture plate.Then they were randomized in-to five groups including control group,blank liposome group( 50 μ1, 100 μ1),clodronate-liposome group (50μ1,100μ1).Values of OD were determined by MTT.AO fluorescence and haematoxylin dye were employed to determine the apoptosis of the AMs.Results There were no significant differences be-tween control group and blank liposome group(50 μ1, 100 μ1).Significant differences were found be-tween control group and clodronate-liposome group(50 μ1, 100 μ1).There were no marked differences between blank liposome group(50μ1, 100 μ1)and clodronate-liposome group(50 μ1,100μ1).AO fluo-rescence and haematoxylin dye were available to define the apoptosis of the AMs.Conclusion Clodr-onate-liposome can effectively induce the apoptosis of the AMs.

OBJECTIVEThe aim of this study was to explore the effect of SPARC on the anti-cancer effect of gemcitabine and underlying mechanism in pancreatic cancer.

METHODSAfter treating with gemcitabine, the proliferation rate of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells was detected by MTT assay. The cell cycle distribution and cell apoptosis in each group were examined by flow cytometry, and the capability of clone formation was tested by adhesion-dependent clone formation assay. The apoptosis-related proteins were analyzed by Western blot.

RESULTSThe growth of pancreatic cancer cells was inhibited by gemcitabine in a time-dependent and dose-dependent manner. Its IC50 at 24, 48, and 72-h was (40.1 ± 2.5) µmol/L, (15.0 ± 0.5) µmol/L and (6.6 ± 0.1) µmol/L, respectively. The overexpression of SPARC increased the inhibitory effect of gemcitabine on growth of pancreatic cancer MIA PaCa2/SPARC69 cells, presenting a dose- and time- dependent manner. Its IC50 at 24, 48, 72 h was (24.3 ± 1.5) µmol/L, (7.7 ± 0.3) µmol/L and (4.8 ± 0.2) µmol/L, respectively. The clone formation assay showed that before gemcitabine treatment, the clone numbers of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (2350 ± 125), (2130 ± 120) and (1567 ± 11), respectively. After gemcitabine treatment, the clone numbers of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were ( 1674 ± 79) , (1587 ± 94) and (557 ± 61), respectively. The overexpression of SPARC enhanced the chemosensitivity of MIA PaCa2 cells to gemcitabine chemotherapy. After treating with 10 µmol/L gemcitabine for 48 h, the ratio of G0/G1 cells in MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (56.0 ± 5.5)%, (55.0 ± 4.5)% and (68.0 ± 7.0)%, respectively. The cells arrested at G0/G1 phase were significantly increased in the MIA PaCa2/SPARC69 cells. The apoptosis rates of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (22.4 ± 2.5)%, (19.9 ± 2.0)% and (37.7 ± 3.9)%, respectively, indicating that overexpression of SPARC enhanced the gemcitabine-induced apoptosis in MIA PaCa2 cells. The Western blot analysis showed that, compared with MIA PaCa2 and MIA PaCa2/V cells, the expression of caspase-2, -8, -9 and cleaved PARP protein was significantly increased, while the expression of Bcl-2 was not changed significantly in the MIA PaCa2/SPARC69 cells.

CONCLUSIONSPARC can enhance the chemosensitivity of pancreatic cancer cells to gemcitabine via regulating the expression of apoptosis-related proteins.

Antimetabolites, Antineoplastic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Caspase 2 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Cycle ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cysteine Endopeptidases ; metabolism ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Humans ; Osteonectin ; metabolism ; Pancreatic Neoplasms ; metabolism ; pathology ; Poly(ADP-ribose) Polymerases ; metabolism ; Time Factors