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As a retrovirus with high recombination and mutation rate， HIV targets CD4+ T lymphocytes， causing damage to the body's immune system and eventually leading to severe immune function defects. Different subtypes of the HIV virus exhibit significant sequence differences in their structural and regulatory genes， and this diversity is closely related to etiology， transmission， diagnosis， drug therapy， disease progression， and vaccine development. As a country with the largest number of HIV subtypes， rapid and accurate typing of the HIV virus holds great significance for China's efforts in disease prevention and control. This study provides a comprehensive reveiw of domestic and international HIV genotyping methods， and summarizes the clinical significance of different subtypes in order to provide reference for further research.
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Objective To investigate the impact of knocking out tumor necrosis factor-related ap-optosis-inducing ligand ( TRAIL) gene ( TRAIL-/-) on colonic inflammation and regulatory T cells ( Treg) in mice with dextran sulfate sodium (DSS)-induced experimental colitis. Methods C57BL/6 mice were ran-domly assigned into four groups with 10 in each group:wild-type ( WT) control, WT colitis, TRAIL-/- con-trol and TRAIL-/- colitis. The mouse model of colitis was induced by oral administration of 3. 5％ DSS and the severity of colonic inflammation was assessed. Peripheral blood mononuclear cells ( PBMCs) and mesen-teric lymph nodes ( MLNs) were collected. The ratios of Treg cells to CD4+T cells in PBMCs were detected by flow cytometry. Expression of Treg cell-associated transcription factor (Foxp3) and cytokine (IL-10) at mRNA level was measured by real-time fluorescent quantitative polymerase chain reaction. Western blot and enzyme-linked immunosorbent assay ( ELISA) were used to detect the expression of Foxp3 and IL-10 at pro-tein level, respectively. Results Compared with the WT control group, the WT colitis group showed signif-icantly decreased proportions of Treg cells in PBMCs [(1. 85±0. 38)％ vs (3. 12±0. 69)％, P<0. 05], but increased proportions in MLNs [(11. 79±1. 18)％ vs (6. 24±1. 04)％, P<0. 05]. Compared with the WT mice with colitis, the TRAIL-/- mice with colitis had more severe colonic inflammation and significantly in-creased proportions of Treg cells in PBMCs [(3. 15±0. 64)％ vs (1. 85±0. 38)％, P<0. 05], but de-creased Treg cells in MLNs [(9. 80±0. 50)％ vs (11. 79±1. 18)％, P<0. 05]. Expression of Foxp3 and IL-10 at mRNA and protein levels in PBMCs of the WT mice with colitis was significantly lower than that in the WT control mice [ Foxp3 mRNA: 0. 48 ± 0. 21 vs 1. 06 ± 0. 31, IL-10 mRNA: 0. 23 ± 0. 07 vs 1. 22 ± 0. 38;Foxp3 protein:0. 68±0. 12 vs 1, IL-10 protein:(4. 91± 0. 72) pg/ml vs (21. 86±2. 40) pg/ml;all P<0. 05], while in MLNs, the expression of Foxp3 and IL-10 at mRNA and protein levels was significantly higher than that of the WT control group [Foxp3 mRNA:3. 71±0. 49 vs 1. 03±0. 15, IL-10 mRNA:11. 98 ±6.10 vs 1. 01±0. 31; Foxp3 protein: 1. 60±0. 03 vs 1, IL-10 protein: (1260. 00±18. 02) pg/ml vs (1184. 00±38. 62) pg/ml;all P<0. 05]. Compared with the WT mice with colitis, the TRAIL-/-mice with colitis showed significantly increased expression of Foxp3 and IL-10 at mRNA and protein levels [ Foxp3 mRNA:1. 80±0. 49 vs 0. 48±0. 21, IL-10 mRNA:1. 67±0. 99 vs 0. 23±0. 07;Foxp3 protein:1. 10±0. 01 vs 0. 68±0. 12, IL-10 protein:(31. 33± 25. 02) pg/ml vs (4. 58±3. 73) pg/ml; all P<0. 05], while de-creased expression in MLNs [ Foxp3 mRNA: 0. 49 ± 0. 21 vs 3. 71 ± 0. 49, IL-10 mRNA: 2. 80 ± 1. 82 vs 11. 98±6. 10; Foxp3 protein: 1. 21±0. 12 vs 1. 60±0. 03, IL-10 protein: (1158. 00±26. 48) pg/ml vs (1190. 00±37. 19) pg/ml;all P<0. 05]. Conclusions Knocking out the expression of TRAIL might af-fect the ratios of Treg cells in peripheral blood and MLNs, thereby aggravating the colitis in mice.
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Objective To investigate the correlation between decoy receptor (DcR) 1,DcR2,osteoprotegerin (OPG) gene polymorphisms and susceptibility of Crohn's disease (CD) in Han population in Zhejiang province.Methods From April 2008 to July 2017,at the Department of Gastroenterology of The Second Affiliated Hospital of Wenzhou Medical University,The First Affiliated Hospital of Wenzhou Medical University,Central Hospital of Wenzhou and Renmin Hospital of Wenzhou,285 patients diagnosed as having CD were enrolled,and during the same period 572 healthy individuals who received health checkup at the Second Affiliated Hospital of Wenzhou Medical University were collected as healthy control.The single nucleotide polymorphism (SNP) of DcR1 (rs12549481),DcR2 (rs1133782) and OPG (rs3102735) were examined by SNaPshot technique.An unconditional logistic regression analysis was performed to analyze the differences in each SNP mutation alleles and genotype frequencies between CD group and control group.Furthermore,their correlation with clinicopathological features of CD and the efficacy of corticosteroid and infliximab was also evaluated.Results The frequencies of mutant allele A and genotype GA + AA of DcR2 (rs1133782) of CD group were 11.93％ (68/570) and 22.81％ (65/285),respectively,which were higher than those of healthy control group (8.22％,94/1 144;and 15.91％,91/572;odds ratio (OR) =1.513,95％ confidence interval (CI) 1.088 to 2.104,P =0.013;OR =1.562,95％ CI 1.094 to 2.230,P =0.014).However there was no statistically significant difference in the mutant allele and genotype frequencies of DcR1 (rs12549481) and OPG (rs3102735) between two groups (all P ＞ 0.05).The frequencies of mutant allele C and genotype TC + CC of DcR1 (rs12549481) in patients with stricturing CD were 13.89％ (25/180)and 27.78％ (25/90),respectively,which were lower than those of patients with non-stricturing,non-penetrating CD (27.68％,62/224 and 48.21％,54/112),and the differences were statistically significant (OR =0.421,95％ CI 0.252 to 0.705,P =0.001;OR =0.413,95％ CI 0.229 to 0.747,P =0.003).Besides,the frequencies of mutant allele A and genotypes GA + AA of DcR2 in patients with penetrating CD were 7.23％ (12/166) and 13.25％ (11/83),which were lower than those of patients with non-stricturing,non-penetrating CD (15.62％,35/224 and 30.36％,34/112),and the differences were statistically significant (OR =0.407,95％ CI 0.205 to 0.809,P =0.009;OR =0.350,95％ CI 0.165 to 0.743,P =0.005).In addition,there was no statistically significant difference in the frequencies of mutant allele and genotypes of OPG (rs3102735) among subtypes of CD with different features (all P ＞ 0.012 5).Moreover,the DcR1 (rs12549481),DcR2 (rs1133782) and OPG (rs3102735) polymorphisms were not correlated with the efficacy of corticosteroid and infliximab (all P ＞ 0.05).Conclusions DcR1 (rs12549481) mutation may be correlated with stricturing CD.DcR2 (rs1133782) mutation may be correlated with CD,especially with penetrating CD.However,the gene polymorphism of OPG (rs3102735) is not correlated with the risk of CD susceptibility.And the above gene SNP may be independent of the efficacy of corticosteroid and infliximab.