Objective: To explore the expression of GATA4 and alpha fetoprotein (AFP) in fetal liver and hepatocarcinoma cells and their correlation. Methods: Semi-quantitive RT-PCR was used to detect the expression of GATA4 and AFP in liver tissues from mouse of E 10.5 d to P 0.5 d, normal liver cells, and hepatocarcinoma cells. Eukaryotic expression vector of mouse GATA4 was used to transfect normal liver cell line and the product was identified. Doxorubicine was used to treat hepatocarcinoma cells; then semi-quantitive RT-PCR was used to determine the mRNA of GATA4, FTF and AFP; Western blot was used to examine the expression of GATA4 and AFP; and ELISA was used to determine AFP in supernatants. Results: The expression of GATA4 and AFP decreased with the maturation of fetal liver cells and reached adult level 0.5 d after birth. GATA4, FTF and AFP were not detected in normal liver cells but were overexpressed in hepatocarcinoma cells. The expression of FTF and AFP was increased in normal liver cells after transfection of GATA4, and the level of AFP was also increased in the supernatant. GATA4 was sharply decreased in hepatocarcinoma cells within 6 h after doxorubicine treatment; the expression of FTF and AFP was also decreased in supernatants 12 h after doxorubicine treatment (P<0.05). Conclusion: GATA4 is highly expressed in fetal liver and hepatocarcinoma cells,which may contribute to the elevation of AFP expression.

Objective To explore the diagnostic value of serum Cys-C, hs-CRP, and urine mALB for early renal injury in gonty patients. Methods A total of 100 gonty patients and 50 healthy volunteers were divided into 3 groups, acute phase group (n = 50), intermittent group (n= 50), and healthy control group (n = 50) . The serum Cys C,hs-CRP,and urine creatinine were determined by automatic chemistry analyzer and urine mALB by immunofluorometer equipment, and the results were analyzed in each group. Results The results of mALB/Cr(mg/[g • Cr]), hs-CRP(mg/L), and Cys-C(mg/L)in acute phase group were 11.45±3.55, 5.15±1.98, and 2.62±0.83, in intermittent group were 8.96±1.78, 3.28±1.23, and 0.93±0.26, and in healthy volunteer group were 3.89± 0.76, 1.91±0.57, and 0.78±0.22, respectively. The values of the above three parameters in the acute phase and intermittent groups were significantly higher than those in the healthy control group (P< 0.05) ; and the values in the acute phase group were significantly higher than those in the intermittent group (P<0.05). The abnormal rate of hs-CRP+ Cys-C+ mALB/Cr was 87% in gonty patients. Conclusion Simultaneous determination of mALB/ Cr, hs-CRP, and Cys-C can help to assess the early renal injury in gonty patients.

Animals ; Ca(2+) Mg(2+)-ATPase ; metabolism ; Ischemic Preconditioning, Myocardial ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

OBJECTIVETo investigate the efficacy of PEG-interferon alpha (PEG-IFN alpha) treatment of HBeAg-positive chronic hepatitis B and HBV genotypes and liver tissues effect of HBeAg seroconversion.

METHODS54 cases confirmed by liver biopsy, genotype clear HBeAg positive chronic hepatitis B (CHB) patients according to body weight, respectively, subcutaneous injection of PEG-IFN-alpha2a 135 microg or 180 microg, or PEG-IFN-alpha2b 50 microg, 80 microg or 100 microg once weekly treatment for 48 weeks and followed for 24 weeks after discontinuation. Statistics of HBeAg seroconvertion, HBV genoty pes and liver histology e antigen seroconversion after the end of treatment.

RESULTS54 patients were followed up at the end of HBeAg seroconversion rate was 29.63% (16/54). Genotype B patients with HBeAg seroconversion rate was 35.29%, 27.03% higher than the C-type patients, but the difference was not statistically significant (chi2 = 0.382, P = 0.537). Inflammation of the liver activity highter ( > G2) , the degree of fibrosis heavier ( > S1) HBeAg seroconversion rate (50.00% vs. 25.00%, 40.90% vs. 21.88%), but were not statistically significant (chi2 = 1.391, 1.444, P = 0.238, 0.229). Activity of HBV genotype, liver inflammation, liver fibrosis and other factors by multivariate Logistic regression analysis, only liver inflammation activity of the important factors of HBeAg seroconversion.

CONCLUSIONImportant factors, liver inflammation activity of PEG-interferon alpha treatment of HBeAg-position chronic hepatitis B patients and HBV genotypes and liver fibrosis may be of little significance.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Genotype ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; pathology ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Logistic Models ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use

OBJECTIVETo evaluate the expression of microRNA (miR)-let-7b in peripheral blood cells of patients with primary biliary cirrhosis (PBC) and investigate its relationship to clinical disease parameters.

METHODSPeripheral blood and serum samples were obtained for study from 60 PBC patients and 60 healthy controls. Peripheral blood cells were extracted and subjected to real-time PCR to measure miR-let-7b expression. Serum levels of interleukin (IL)-18, total bilirubin (TBIL), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) were measured by standard biochemical assays. The relationship between miR-let-7b expression and disease parameters was assessed by Spearman's rank correlation test.

RESULTSPBC patients showed significantly lower expression of miR-let-7b in peripheral blood cells than healthy controls (P less than 0.001); moreover, the miR-let-7b expression level decreased in parallel to increases in disease severity (stage I > II / III > IV). In PBC patients, the miR-let-7b expression was significantly correlated with Mayo risk scores (r = -0.4930, P less than 0.001), IL-18 (r = -0.4643, P less than 0.001) and ALP (r = -4119, P less than 0.001), but not with TBIL or GGT.

CONCLUSIONDecreased expression of miR-let-7b may be associated with development and progression of PBC, and this miRNA may represent a novel target of improved diagnostic and preventive strategies for PBC.

Adult ; Aged ; Alkaline Phosphatase ; blood ; Bilirubin ; blood ; Case-Control Studies ; Female ; Humans ; Interleukin-18 ; blood ; Liver Cirrhosis, Biliary ; blood ; Male ; MicroRNAs ; metabolism ; Middle Aged ; gamma-Glutamyltransferase ; blood

OBJECTIVETo investigate relationship between glucose metabolic disorders and expression of insulin receptor (IR) and tyrosine protein kinase (TPK) in posthepatitic cirrhosis hepatocyte and HBV DNA expression in pancreatic cells.

METHODSTo detect HBV DNA in paraffin-embedded pancreatic and hepatic tissues from 12 posthepatitic cirrhosis patients with positive serum HBV markers by using in situ hybridization (ISH) with a digoxigenin labelled probe. The amount of IR and TPK have been evaluated by immunohistochemical quantitative analysis using image analyzer in hepatocyte of 12 patients positive for HBV markers with posthepatitic cirrhosis in serum. Immunofluorescent histochemical double staining technique was used. HBsAg and IR were observed under confocal laser scanning microscope.

RESULTSEleven of 12 cirrhosis patients? hepatocytes were HBV DNA positive, including 7 patients (7/7) with impaired glucose tolerance (IGT) and 4 patients (4/5) with normal glucose tolerance (NGT). Eight of 12 pancreatic cells were HBV DNA positive, including 7 patients (7/7) with IGT, but only one patient (1/5) with NGT-HBV DNA was found positive in pancreatic cells in significantly more subjects in IGT group than in NGT group (P less than 0.01).IR and TPK amount in hepatocyte of IGT was significantly less than that of NGT patients with posthepatitic cirrhosis (P less than 0.01). IR amount was closely related to the TPK in cirrhosis hepatocyte r=0.82597(P less than 0.01). HBV DNA was mainly localized in the nuclei of hepatocyte and pancreatic acinar and islet cells. Immunofluorescent histochemical double-staining showed that HBsAg was partly localized in the IR positive areas of hepatocytes and pancreatic islet cells.

CONCLUSIONHBV can invade acinar cells of pancreas and islet cells, which might be a direct cause of insulin-dependent diabetes mellitus-like the disorder and insulin absence after HBV infection. Decrease of IR and TPK might be main cause of noninsulin-dependent diabetes mellitus-like disorder after having hepatitis or posthepatitic cirrhosis.

DNA, Viral ; analysis ; Female ; Glucose Metabolism Disorders ; complications ; metabolism ; virology ; Hepatitis B virus ; genetics ; Hepatocytes ; metabolism ; virology ; Humans ; In Situ Hybridization ; Liver Cirrhosis ; complications ; metabolism ; virology ; Male ; Middle Aged ; Pancreas ; cytology ; virology ; Protein-Tyrosine Kinases ; metabolism ; Receptor, Insulin ; metabolism

OBJECTIVETo understand and master the situation in which enterovirus caused hand-foot-and-mouth disease (HFMD) in Xuzhou district in 2009 so as to provide scientific basis for the control and prevention of hand-foot-and-mouth disease.

METHODSThe researchers adopted fluorescence RT-PCR method to detect EV and EV71 as well as the CA16 specificity RNA from 222 samples of anal swabs and oropharyngeal swabs from the 240 cases who were diagnosed clinically as hand-foot-mouth disease infected by enterovirus. Also, the researchers conducted EV71-IgM antibody detection on 114 samples of acute phase serum with ELA method.

RESULTSAmong the 240 enterovirus infected patients, the total EV infection rate is 72.50% (174/240), among which EV71 infection rate is 57.92% (139/240), CoxA16 infection rate is 9.17% (22/240), and other EV infection rate is 5.42% (13/240). The EV71-RNA positive rate of the samples of 222 anus swabs among the 240 suspected enterovirus infected patients is 45.94% (102/222), the samples of swallow swab EV71-RNA positive rate is 25.68% (57/222) and the EV71-IgM antibody positive rate of 114 samples of acute phase serum is 86.84% (99/114). The EV71-RNA positive rate of oropharyngeal swabs of 254 healthy children is 1.57% (4/254) , and no CoxA16-RNA was detected. In the oropharyngeal swabs of 54 close contacts (medical personnel), the EV-RNA detected is negative. The positive rate of EV71-IgM antibody of the 258 healthy children's serum samples is 2.71% (7/258).

CONCLUSIONThe widespreading of hand-foot-mouth disease in Xuzhou district is caused mainly by type 71 enterovirus. Inapparent infection of type 71 enterovirus exists among children under the age of 3 during the time of widespreading period and IgM antibody develops in them. It is difficult for adults to be infected by EV71 even if they contact the contagion source closely. The positive rate of EV71-IgM antibody in the samples of acute phase serum of suspected cases is the highest (86.84%), and the second highest is the positive rate of RNA of EV71 of anal swabs (45.94%) and of the EV71 of oropharyngeal swabs (25.68%). ELA reagent kit is used in the early diagnosis of EV71 infection for it is easy to operate, fast and economic, so, it is worth popularizing in the grass-root medical units.

Child, Preschool ; China ; Enterovirus A, Human ; genetics ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Male

OBJECTIVEIn order to provide a scientific basis for influenza prevention and control, analyzing the epidemic characteristics and laws of influenza outbreaks in Xuzhou area during 2005-2011.

METHODUsing fluorescent-PCR method to detect influenza virus nucleic acid on Nasopharyngeal swab specimens collected from influenza outbreak cases during 2005-2011 and fast classifying influenza virus A1 (H1N1), A3 (H3N2), new H1N1 BV (Victoria) and BY (Yamagate) on subtypes. At the same time, isolating the influenza virus with MDCK cells, and sending them to the National Influenza Center for review, after the preliminary identification of the isolated influenza virus.

RESULTSDuring 2005-2011, there are 53 influenza outbreaks in Xuzhou area, which caused by influenza virus subtype BV accounting for 26.42% (14/53), A3 accounting for 49.1% (26/53), A3 and A1 mixture accounting for 3.77% (2/53) and the new H1N1 accounting for 20.75% (11/53). The outbreaks in 2007 and 2009 mainly caused by A3, and show that the winter spring (January) and summer autumn (September) as two popular peaks during 2005-2011; BV mainly causes the outbreaks from Feb. to Jun.

CONCLUSIONIn Xuzhou area, since the winter of 2005, influenza virus subtype BV, the A3, and new H1N1 has alternately as mainly predominant strain, caused local influenza outbreaks. In which BV has increased trend year by year during 2005-2011. The students in primary and secondary schools are the major crowd of influenza outbreaks. Fluorescent-PCR detection methods could be a preferred method for reliable and rapid diagnostic of epidemic influenza outbreaks.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Male ; Middle Aged ; Public Health Surveillance ; Seasons ; Young Adult

OBJECTIVETo investigate the prevalence and subtypes of influenza viruses in Xuzhou city from 2005 to 2011 and to provide the scientific supports for influenza prevention and control in this religion.

METHODSThe throat swab samples were collected from the influenza-like cases from national influenza like illness sentinel hospital in Xuzhou. The samples were used for influenza virus isolation and identification, sent on the national flu center to confirm according to the "national influenza surveillance program" and "influenza virus and experimental technology".

RESULTSFrom Oct. 2005 to Dec. 2011, a total of 9561 swab specimens were collected in which 1152 strains were identified for influenza viruses with total isolated rate of 12.0%. Among these strains, 708 strains were A1 (H1N1) subtype (14.2%), 466 strains were A3 (H3N2) subtype (40.5%), 78 strains were new H1N1 subtype (6.8%), 362 strains were BV (Victoia) subtype (31.4%) and 82 strains were BY (Yamagate) subtype (7.1%). The top detection rate (25.9%) arose in 2007, secondary detection rate (17.4%) occurred at 2009 and the lowest one (2.3%) appeared in 2011. From the winter of 2005 to the spring of 2006 A1 (H1N1) subtype had appeared as predominant strains but in the winter of 2006 the predominant strains were BV subtype. It changed to A3 subtype in 2007 to 2009 and the other three dominant strains were A1, BV and BY in 2008. In the winter of 2009, both A3 (H3N2) and new H1N1 subtype were predominant strains. BV subtype was predominant strains in 2010 to 2011. The prevalence of A3 subtype appeared in all the year while prevalence of BV only arose in the spring and winter. So the detection rate was high in January (34.4%) but low in August (2.2%). The influenza population is correlated with age, the highest detection rate arose in 5-age group and the lowest detection rate appeared in 25-age group.

CONCLUSIONInfluenza subtype A1, A3, New H1N1 are all appeared as predominant strains in Xuzhou city from 2005 to 2010. Besides, the prevalence of BV subtype is stronger in recently.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Male ; Middle Aged ; Public Health Surveillance ; Seasons ; Young Adult