Could Intrathymic Injection of Myelin Basic Protein Suppress Inflammatory Response After Co-culture of T Lymphocytes and BV-2 Microglia Cells? / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The interaction between activated microglia and T lymphocytes can yield abundant pro-inflammatory cytokines. Our previous study proved that thymus immune tolerance could alleviate the inflammatory response. This study aimed to investigate whether intrathymic injection of myelin basic protein (MBP) in mice could suppress the inflammatory response after co-culture of T lymphocytes and BV-2 microglia cells.</p><p><b>METHODS</b>Totally, 72 male C57BL/6 mice were randomly assigned to three groups (n = 24 in each): Group A: intrathymic injection of 100 μl MBP (1 mg/ml); Group B: intrathymic injection of 100 μl phosphate-buffered saline (PBS); and Group C: sham operation group. Every eight mice in each group were sacrificed to obtain the spleen at postoperative days 3, 7, and 14, respectively. T lymphocytes those were extracted and purified from the spleens were then co-cultured with activated BV-2 microglia cells at a proportion of 1:2 in the medium containing MBP for 3 days. After identified the T lymphocytes by CD3, surface antigens of T lymphocytes (CD4, CD8, CD152, and CD154) and BV-2 microglia cells (CD45 and CD54) were detected by flow cytometry. The expressions of pro-inflammatory factors of BV-2 microglia cells (interleukin [IL]-1β, tumor necrosis factor-α [TNF-α], and inducible nitric oxide synthase [iNOS]) were detected by quantitative real-time polymerase chain reaction (PCR). One-way analysis of variance (ANOVA) and the least significant difference test were used for data analysis.</p><p><b>RESULTS</b>The levels of CD152 in Group A showed an upward trend from the 3rd to 7th day, with a downward trend from the 7th to 14th day (20.12 ± 0.71%, 30.71 ± 1.14%, 13.50 ± 0.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The levels of CD154 in Group A showed a downward trend from the 3rd to 7th day, with an upward trend from the 7th to 14th day (10.00 ± 0.23%, 5.28 ± 0.69%, 14.67 ± 2.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The ratio of CD4+/CD8 + T in Group A showed a downward trend from the 3rd to 7th day, with the minimum at postoperative day 7, then an upward trend from the 7th to 14th day (P < 0.05). Meanwhile, the levels of CD45 and CD54 in Group A were found as the same trend as the ratio of CD4+/CD8 + T (CD45: 83.39 ± 2.56%, 82.74 ± 2.09%, 87.56 ± 2.11%; CD54: 3.80 ± 0.24%, 0.94 ± 0.40%, 3.41 ± 0.33% at postoperative days 3, 7, and 14, respectively, P < 0.05). The expressions of TNF-α, IL-1β, and iNOS in Group A were significantly lower than those in Groups B and C, and the values at postoperative day 7 were the lowest compared with those at postoperative days 3 and 14 (P < 0.05). No significant difference was found between Groups B and C.</p><p><b>CONCLUSIONS</b>Intrathymic injection of MBP could suppress the immune reaction that might reduce the secondary immune injury of brain tissue induced by an inflammatory response.</p>

Association between IL-10 gene polymorphisms and the outcomes of hepatitis C virus infection among high-risk populations in Jiangsu province / 中华流行病学杂志

Objective To explore the relationship between interleukin (IL)-10 gene polymorphisms and the susceptibility or the outcomes of HCV infection among high-risk populations in Jiangsu province.Methods IL-10 gene SNPs were detected in 1555 subjects including 264 self-limited HCV infections.371 persistent HCV infections and 920 healthy controls were selected through Taqman-MGB.Results After adjusted for cofounders as sex,age and high-risk population,data from logistic regression analysis showed that the distribution of IL-10 genotypes among the controls,spontaneous clearances and those with persistent infections did not show much differences.Results from further stratified analysis showed that,at the position of-819T/C,when compared with TT genotype,TC genotype had a significantly increasing chance of self-limited HCV infection among middle-aged,females and paid blood doners (adjusted OR values and 95％ CI were:2.160,1.163 4.011 ；1.693,1.066-2.688 and 4.084,1.743-9.570).It also had a lower risk of progressing to persistent HCV infection among those paid blood doners (the adjusted OR values and 95％CI were:0.312,0.130-0.747 ).CC genotype had a higher chance of self-limited HCV infection among people underwent blood dialysis (the adjusted OR values and 95％CI were:2.120,1.071 -4.197).Results also showed a decreased risk of progressing to persistent infection among paid blood doners (the adjusted OR values and 95％CI were:0.156,0.043-0.566).At the position of -592A/C,when compared to AA genotypc,the AC genotype had a significantly increasing chance of self-limited HCV infection among middle-aged,femalcs and paid blood doners (the adjusted OR values and 95％ CI were:2.176,1.173-4.037；1.659,1.055-2.607;3.704,1.625-8.443) but had an increased risk of persistent HCV infection among females (the adjusted OR values and 95％CI were:1.525,1.017-2.286).AC genotype showed an increased opportunity to progress to HCV persistent infection among drug users (the adjusted OR values and 95％CI were:1.845,1.122-3.034) but had a reduced risk of progressing to HCV persistent infection among paid blood doners (the adjusted OR values and 95％CI were:0.361,0.155-0.841 ).CC genotype had an increased opportunity to self-linited HCV infection as well as having a dccreased risk of progressing to persistent infection among paid blood donets (the adjusted OR values and 95％CI were:3.125,1.016-9.605；0.218,0.063-0.748).At the position of-1082A/G,AG/GG genotypcs had an increased chance of self-limited infection among blood doners (the adjusted OR values and 95％CI were:3.780,1.620-8.820).Conclusion IL-10-819T/C,-592A/C,-1082A/G SNPs might be related with the susceptibility and the outcomes of HCV infection among populations at high risk.

Influence of F protein of hepatitis C virus subtype 1b inhibits on human hepatocellular carcinom HepG2 cell apoptosis / 中华流行病学杂志

Objective To investigate the effects of F protein of hepatitis C virus subtype lb on the apoptosis of human hepatocellular carcinom HepG2 cells. Methods HepG2 cells were transfected with recombinant plasmid pcDNA3.0-F-EGFP and pcDNA3.0-F-EGFP-HepG2 strain was exposed to Act-D and tumor necrosis factor a (TNFα) treatment in order to induce cell apoptosis with positive control pcDNA3.0-C-EGFP-HepG2, negative control pcDNA3.0-C-EGFP-HepG2 and blank control HepG2.Annexin V-FITC/PI of flow cytometry was performed to determine the number of apoptotic cells. DNA Ladder was used to observe the isolation of apoptotic DNA fragments in the apoptotie cells. Results pcDNA3.0-F-EGFP- HepG2 cell strain showed a much delayed apoptosis as well as obviously lowering the apoptotic rate when compared with the pcDNA3.0-HepG2 strain and HepG2 strain (P<0.001).Conclusion The introduction and expression of extraneous gene (the F gene of hepatitis C virus subtype 1b) could significantly inhibit the apoptosis of HepG2 cells.

Detection of serum anti-F antibody in hepatitis C virus infected patients / 中华流行病学杂志

<p><b>OBJECTIVE</b>To assess the prevalence of serum anti-F in patients with hepatitis C virus (HCV) infection and the distribution of anti-F.</p><p><b>METHODS</b>The recombinant protein (HCV-F/GST) was coated onto micro titer plates as antigen. Sera of 120 patients with hepatitis C virus infection, 15 patients with hepatitis B, 3 patients with hepatitis E and 10 normal sera were tested by indirect ELISA for detecting anti-F.</p><p><b>RESULTS</b>82 samples out of the 120 (68%) HCV infected patients exhibited a positive anti-F reaction, showing significant difference from the controls with no HCV infection (P < 0.01). Data from logistic analysis showed that the positive rate of anti-F was higher in patients over 50 year olds (OR = 6.675, 95% CI: 2.407-19.071). Patients of midrange, severe phase and hepatic cirrhosis had higher rate than the others (OR = 2.749, 95% CI: 1.470-5.141).</p><p><b>CONCLUSION</b>Prevalence and distribution of anti-F in Yixing hepatitis C patients was reported and which might be related to the progression of HCV infection.</p>

The genetic polymorphism of HLA-DQA1 and HLA-DQB1 genes of Chinese Han population in Jiangsu area is studied by PCR-sequence-based typing / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the polymorphism of HLA-DQA1 and DQB1 genes of Han population in Jiangsu of China.</p><p><b>METHODS</b>The alleles and haplotypes frequencies of HLA-DQA1 and DQB1 genes in 100 unrelated healthy individuals were analyzed by using polymerase chain reaction-sequence-based typing (PCR-SBT).</p><p><b>RESULTS</b>Among the 7 DQA1 alleles detected, the most common allele was DQA1*0301/02/03 with a frequency of 29.5%, which was followed by DQA1*0501, DQA1*0102 and DQA1*0201 with frequencies of 18.5%, 17.0% and 12.5%, respectively. Of the 13 DQB1 alleles detected, DQB1*0201/02 allele (21.5%) was the most frequent allele, followed by DQB1*0301/09 (14.5%), DQB1*0303 (13.5%) and DQB1*0603 (11.5%). The most common DQA1 vs DQB1 haplotype was DQA1*0301/02/03 vs DQB1*0303 with a frequency of 12.5%, which was followed by the DQA1*0201-DQB1*0201/02 (10.5%),DQA1*0501-DQB1*0201/02 (9.5%) and DQA1*0501-DQB1*0301/09 (7.0%).</p><p><b>CONCLUSION</b>The distribution of HLA-DQ alleles and haplotypes in Jiangsu Han population shares some genetic characteristics with other population in northern of China, but has its own characteristics. The data will provide useful information for anthropology, organ transplantation and disease association studies.</p>

Study on hepatitis C virus genotyping in Yixing area, Jiangsu province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the distribution of hepatitis C virus (HCV) genotypes in Yixing, Jiangsu province.</p><p><b>METHODS</b>Genotypes identification on sera samples were obtained from 158 donors who had already been anti-HCV positive through PCR method with type specific primer designed according to the sequence of 5'non-coding region (5'NCR). 5'NCR was also sequenced and compared with published date. Genotypes distribution was investigated in patients with different sex and clinical types of hepatitis C.</p><p><b>RESULTS</b>Of the total 158 patients, 95 were HCV RNA positive in which 80 patients having genotype 1b (80/95; 84.4%), 5 patients having genotype 2(5/95; 5.3%), 5 patients with 1b/2 mixed genotypes (5/ 95; 5.3%) and another 5 patients whose genotype undetermined. The difference on the distribution of HCV genotypes was significant between female and male patients (P < 0.05) but not in different kinds of hepatitis C patients.</p><p><b>CONCLUSION</b>Type 1b was the predominant HCV genotype in Yixing area.</p>