Emerging data indicated that HCV subverts the antiviral activity of interferon (IF); however,whether HCV core protein contributes to the process remains controversial. In the present study, we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway. Our results showed that, following treatment with IFN-α, the transcription of PKR, MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein. In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased. Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1, whereas the level of STAT1 expression was unchanged.Accordingly, SOCS3, the negative regulator of the Jak/STAT pathway, was induced by HCV core protein. These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.

OBJECTIVETo study the relationship between influenza epidemic and genetic characteristic on HA and NA regions of influenza virus subtype A3 isolates of Zhejiang province in the recent years.

METHODSRNA of 25 influenza virus subtype A3 isolates, circulated in Zhejiang province during 1998 to 2005, was extracted. HA1 and NA regions were amplified and sequenced. All the sequence data were analyzed using BioEdit.

RESULTSHA1 and NA regions of all the isolates belonged to 987nt and 1362nt, encoding protein of 329 and 454 amino acids respectively. Isolates shared amino acid homology of 90.9%-99.3% and 95.2%-99.5% on HA1 and NA regions, while divergence on HA1 was greater than that on NA region. During a period of 8 years, 30 amino acids on HA1 region were substituted and 14 of which refer to 4 antigenic determinant sites. Meanwhile,21 amino acids on NA region were substituted and 5 of which referred to 3 antigenic determinant sites. Significant divergences, both in HA1 and NA, were observed among isolates in 1998 and 2002, showing that they belonged to absolutely different branches. Additionally, influenza virus subtype A3 isolates identified in recent years, with 11 N-linked glyeosylation sites in HA1 region, had 5 sites more than early A/Aichi/2/68 strain. Since 1998,3 sites had been inserted in epidemic strains, indicating the accelerated trend of glyeosylation sites were increasing.

CONCLUSIONThere is a correlation between antigenic drift of influenza virus subtype A3 and the two epidemics in Zhejiang province in 1998 and 2002.

Amino Acid Sequence ; Antigens, Viral ; genetics ; China ; Epitopes ; genetics ; Evolution, Molecular ; Genetic Drift ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; Molecular Sequence Data ; Neuraminidase ; genetics ; RNA, Viral ; genetics ; Sequence Analysis, RNA ; Sequence Homology, Amino Acid

Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.

Objective To construct PPAR?and PPAR?response element (PPRE)-controlled luciferase expression vectors,and to determine whether the traditional Chinese medicine emodin activates PPAR?and improves the glucose uptake by HepG2 hepatocytes.Methods (1) PPAR?and PPRE luciferase expression vectors were constructed and were applied to screen more than 20 ingredients of the traditional Chinese medicine. (2) HepG2 cells were incubated with emodin which can activate PPAR?and PPRE luciferase activity,and the PPAR?mRNA expression level was evaluated by RT-PCR/Southern blot.(3) PPAR?and glucose transporter 2 (Glut2) proteins were determined by Western blot analysis in HepG2 cells treated with emodin.(4) The glucose uptake rate was measured using 2-deoxy-[~3H]-D-glucose in HepG2 cells after treatment with emodin.Results (1) Emodin stimulated luciferase activity controlled by PPRE in dose-dependent manner at concentrations of 0.04 to 180?mol/L in COS-7 cells.The highest value was about 4 folds of control in the cells treated with 90?mol/L emodin (P

OBJECTIVETo study the differences between epidemic strains of influenza virus subtype A3 circulated in China and Occident in past 18 years, in genetic level, and vaccine strains recommended by WHO in corresponding time.

METHODSAmino acid sequences of HA1 regions of the epidemic strains, which circulated in China and Occident from 1988 to 2005, and the vaccine strains of influenza virus subtype A3 were compared by BioEdit and analyzed on the differences of HA1 and it's antigen determinants

RESULTSDifferences between epidemic strains and vaccine strains recommended in corresponding year, both in HA1 and it' s antigen determinant regions, were obviously greater than that between epidemic strains and vaccine strains recommended in next round (P< 0.01). However, differences between epidemic strains in Occident and vaccine strains recommended in corresponding year were slightly greater than that between epidemic strains and vaccine strains recommended in next round and it was not marked (P >0.05). In addition, differences between epidemic strains and vaccine strains which being used for several years, whether in China or in Occident, constantly increased accompanying the used time prolonged.

CONCLUSIONThere was an obvious lag between vaccine strains recommended by WHO, analyzed in genetic level, and epidemic strains of influenza virus subtype A3 circulated in China.

Antigens, Viral ; analysis ; China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; Humans ; Influenza A virus ; classification ; Influenza Vaccines ; Influenza, Human ; classification ; epidemiology ; prevention & control

OBJECTIVETo explore the neutralization capacities of different types of human serum to measles virus epidemic strains and vaccine strain.

METHODSNeutralization antibody (NT) to Shanghai 191 and measles virus isolates in 2005 were tested using acute and convalescent serum samples from diagnosed measles patients, children serum samples collected before and after vaccination and serum samples of migrant residents, from 3 different regions. Additionally, animal immune serum referring to vaccine strain and 3 epidemic strains were prepared and used to undergo crossing neutralization test with corresponding strains mentioned-above. Antigenic ratios were calculated.

RESULTSGMT value of NT of after-immune serum to vaccine strains was 50.82,1.86 times higher than that to MVi/ZJ/05/7 (GMT was 27.35), whereas GMT value of convalescent serum to MVi/ZJ/05/7 (GMT was 386.95) was obviously higher than that to vaccine strain (GMT was 1:151.83),and GMT value of migrant residents' serum in 3 regions to MVi/ZJ/05/7 were 2.22-4.17 times lower than that to vaccine strain. Meanwhile,the antigenic ratios between MVi/ZJ/ 99/1, MVi/ZJ/04/1, MVi/ZJ/05/7 and vaccine strain were found to be 4.28,5.24 and 5.66 respectively. Additionally,low NT titers to vaccine strain were found in patients' acute sera and GMT value was over 1:4.

CONCLUSIONThere were obvious differences on neutralization antibody of different types of serum to measles vaccine strain and epidemic strains which indicating the antigenic diversity of epidemic strains had influenced the protective effectiveness of vaccine antibody to epidemic strains. It was of significance to carry on research projects on the antigenic diversity and effectiveness of measles vaccine.

Animals ; Antibodies, Neutralizing ; blood ; immunology ; Antibodies, Viral ; blood ; immunology ; Antigens, Viral ; genetics ; immunology ; Child ; China ; epidemiology ; Humans ; Measles ; epidemiology ; immunology ; prevention & control ; Measles Vaccine ; immunology ; Measles virus ; genetics ; Neutralization Tests ; Vaccination

Objective To analyze the genetic characteristics of the complete sequence of coxsackievirus A24 variant(CA24v) isolated from acute hemorrhagic conjunctivitis (AHC) outbreaks in Zhejiang province during 2002 to 2010.Methods Complete sequences of CA24v epidemic strains isolated in different years were amplified under the RT-PCR assay,while the sequences of whole genome,VP1,and 3C region of Zhejiang strains were compared with epidemic strains isolated in other areas of China and abroad.Results The whole genome of Zhejiang CA24v strains isolated in 2002 and 2010 was 7456-7458 bp in length,encoding a polyglutamine protein which containing 2214 amino acid residues.There was a insertion with T on site 97 and 119 within 5' non-coding region between epidemic strain Zhejiang/08/10 and strains isolated in 2002.The rates of amino acid homology among Zhejiang/08/10 and other strains isolated since 2002 were between 94.7％ and 100.0％.Compared with the representative strains circulated within the recent 60 years,the largest average amino acid variations had been occurred on region 2A and 3A,with the ratios as 8.4％ and 7.3％ respectively.The smallest variation happened in region 3D,with the ratio only as 1.9％.The rates of stable amino acid variation on the whole genome between strains isolated since 1987 and 2002 were 38 and 20.P-distance within groups appeared that region 3C was more stable than VP1 of strains isolated in 2002-2010,and the 3D of early strain Jamaica/10628/87 might have had a nature of recombination but not observed on those epidemic strains in recent years.Conclusion Within the evolution of CA24v strains,the time course was more significant than the geographical differences.There had been sporadic epidemics of AHC caused by CA24v in Zhejiang province since 2002.

To observe the serum samples and the anti-inflammatory effect of Tripterygium wilfordii in treating RA by using the pharmacokinetic-pharmacodynamic model, make a correlation analysis on concentration-time and effect-time curves, and explore RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in rats by PCR. Methotrexate, tripterine and high-dose T. wilfordii could down-regulate RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in AA rat lymph nodes. The study on PK-PD model showed correlations between inflammatory factors and blood concentration of T. wilfordii. T. wilfordii and its main active constituent tripterine could show the inflammatory effect and treat RA by inhibiting IL-17 cytokine.
Animals ; Arthritis, Rheumatoid ; drug therapy ; immunology ; Biomarkers ; Female ; Interleukin-17 ; antagonists & inhibitors ; genetics ; Interleukin-6 ; genetics ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Tripterygium ; Triterpenes ; pharmacokinetics ; pharmacology

OBJECTIVETo explore the active ingredients in the Chinese yellow wine could inhibit the proliferation and migration of rat vascular smooth muscle cells induced by homocysteine (Hcy).

METHODSThe primary culture and identification of rat vascular smooth muscle cells (VSMCs) was conducted, and the VSMCs in passage 4-7 were used in the following experiments. The VSMCs were divided into 7 groups: control, Hcy (1 mmol/L), Hcy + oligosaccharide, Hcy + polypeptides, Hcy + polyphenols, Hcy + alcohol, Hcy + Chinese yellow wine and were given the corresponding treatment. The proliferation of VSMCs was determined by MTT. Transwell chambers and would healing were employed to test the migratory ability of VSMCs. Wester blot and gelatin zymography were used to investigate the expressions and activities of metal matrix proteinase 2/9 (MMP-2/9) and tissue inhibitor of metalloproteinase 2 (TIMP-2) in VSMCs of each group.

RESULTSCompared with control group, the proliferation, migration and the expression and activity of MMP-2/9 of VSMCs were significantly increased in the VSMCs of Hcy group (P < 0.01). Compared with Hcy group, the proliferation, migration and the expression and activity of MMP-2/9 of VSMCs were significantly decreases in the VSMCs of polypeptides group, polyphenols group and Chinese yellow wine group. However, the expression of TIMP-2 among each group had no significant difference.

CONCLUSIONPolypeptides and polyphenols in the Chinese yellow wine could inhibit the proliferation and migration of VSMCs induced by Hcy.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Homocysteine ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Peptides ; chemistry ; Polyphenols ; chemistry ; Rats ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Wine

OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology