Rheumatoid arthritis (RA) is caused by complicated interactions between genes and the environment. CXC chemokine receptor 5 (CXCR5) is required for B and T follicular helper cell migration and humoral immunity generation. Therefore, this study aimed to assess whether polymorphisms of the CXCR5 gene are implicated in RA development and progression. This case-control study enrolled 285 RA patients and 291 healthy controls. The polymerase chain reaction-ligase detection reaction method was used to genotype rs630923, rs497916, rs3922, and rs676925 in the CXCR5 gene. Epidemiological, clinical, and laboratory data were collected retrospectively. The rs630923 A allele was associated with a higher risk of RA (AOR [adjusted odds ratio]=2.00, 95% confidence interval [CI] =1.14-3.53). However, in the RA group, the frequency of the rs497916 T allele was lower (AOR=0.69, 95% CI=0.51-0.93). Regarding rs3922, AG+GG genotype carriers were at a significantly lower risk for RA than AA genotype carriers (AOR=0.70, 95% CI=0.49-0.99). In the RA group, we found that the different genotypes were significantly associated with specific laboratory values, including rheumatoid factor, total bilirubin, total cholesterol, low-density lipoprotein cholesterol, and alkaline phosphatase. This is the first report indicating that CXCR5 polymorphisms were associated with RA susceptibility. These findings lead to a rising possibility of identifying RA-susceptible individuals based on genetic markers.

Abstract We previously identified E26 transformation specific sequence 1 (ETS1) rs73013527 single nucleotide polymorphism associated with RA susceptibility and disease activity. In the present study, we aims to further investigate the association between ETS1 rs73013527 and receptor activator of nuclear factor kappa B ligand (RANKL), an index related to bone destruction and was reported to elevate in RA. We determined genotypes of ETS1 rs73013527, serum RANKL concentration, clinical characteristics (disease duration, disease activity score for 28 painful/swollen joints), and laboratory markers (rheumatoid factor, anti-citrullinated protein antibody, anti-keratin antibody, c-reactive protein, erythrocyte sedimentation rate) of 254 RA cases. Univariate and multivariate analysis were employed to explore the association between ETS1 rs73013527 and serum RANKL levels in RA patients. Univariate and multivariate analysis indicated no association of serum RANKL levels with patient age, gender, clinical characteristics, and laboratory markers. Univariate analysis, not multivariate analysis indicated genotype CT/TT of ETS1 rs73013527 was significantly associated with elevated RANKL levels in RA patients. ETS1 rs73013527 is in relation to serum RANKL levels among patients with RA. ETS1 probably might be an indirect factors involved in RANKL regulation in RA.

Ikaros family zinc finger 1(IKZF1) encodes a lymphoid-restricted zinc finger transcription factor named Ikaros that regulates lymphocyte differentiation and proliferation as well as self-tolerance. Increasing evidence indicates that IKZF1 could contribute to the pathogenesis of autoimmune diseases. Recent research has provided evidence that IKZF1 might correlate with Systemic lupus erythematosus (SLE), but no clear definition has been made yet. In this study, we focus on the relationship between IKZF1 polymorphisms and SLE susceptibility, cytokine levels, and clinical characteristics in the Chinese Han population. One thousand seventy-six subjects, including 400 SLE patients and 676 healthy controls, were included in this study. Three single nucleotide polymorphisms within IKZF1 containing rs4917014, rs11980379, and rs4132601 were genotyped in all subjects by an improved multiplex ligation detection reaction technique. 143 subjects from SLE patients were randomly selected for testing the levels of serum cytokines. The clinical characteristics of SLE patients were gathered and collated from medical records. The data were analyzed mainly using SPSS20.0 (SPSS lnc., Chicago, IL). Significant relationships were observed between rs4132601 and SLE susceptibility, CD40 ligand, and malar rash (P < .001,P = .04, andP = .01, respectively). In addition, significant relationships were observed between rs4917014 and susceptibility, granzyme B level, and hematological disorder in SLE (P = .005,P = .03 andP = .005, respectively). The results further support that IKZF1 may have an important role in the development and pathogenesis of SLE. Allele G of rs4132601 and rs4917014 is related to a decreased risk of SLE occurrence and associated with clinical features in SLE patients, including CD40 ligand level, granzyme B level, malar rash, and hematological disorder, which play important roles in disease progression.

Tumor necrosis factor-alpha (TNF-alpha) plays an important role in autoimmune diseases. Previous studies have investigated the association of TNF-alpha-238G/A (rs361525) and -308G/A (rs1800629) polymorphisms with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). However, no agreed conclusion had been made. Therefore, this meta-analysis was conducted to assess the associations of TNF-alpha-238G/A and -308G/A polymorphisms with RA and SLE risk. A systematic search was conducted in commonly used databases. Meta-analysis was performed by STATA12.0. A total of 43 studies were included. In the overall population, the TNF-alpha-238A allele was observed to be a protective factor for RA (A vs G: OR=0.75, 95%CI=0.570.99, P=0.040) and the TNF-alpha-308A allele was found to be a risk factor for SLE (A vs G: OR=1.78, 95%CI=1.45-2.19, P <0.001). However, no evidence of association was found between TNF-alpha-238 G/A polymorphism and SLE nor between -308G/A and RA. In the subgroup analysis, TNF-alpha-308A allele played a pathogenic role for RA in Latin Americans (A vs G: OR=1.46, 95%CI=1.15-1.84, P=0.002) and for SLE in Latin Americans (A vs G: OR=2.12, 95%CI=1.32-3.41, P=0.002) and Europeans (A vs G: OR=2.03, 95%CI=1.56-2.63, P <0.001), while it played a protective role for RA in Asians (A vs G: OR=0.54, 95%CI=0.32-0.90, P=0.017). No significant association was found between TNF-alpha-308G/A and SLE susceptibility in Africans and Asians. This meta-analysis demonstrated that TNF-alpha-238A was associated with decreased risk of RA rather than SLE, while -308G/A polymorphism was associated with SLE rather than RA. Stratification analysis indicated that different ethnicities would have different risk.

Although the SLE risk gene loci of HLA-DR and HLA-DQ within the major histocompatibility complex (MHC) region has been gradually revealed by recent Genome-Wide Association studies (GWAS), the association of HLA-DP polymorphisms with SLE was minimally reported. Considering that the variants in rs3077 and rs9277535 in the HLA-DP region could influence the immune response by affecting antigen presentation of HLA class II molecules to CD4+ T cells, the present study aimed to explore the role of HLA-DP polymorphisms in SLE. In total, samples from 335 SLE patients and 635 healthy controls were collected and genotyped by a polymerase chain reaction-high resolution melting (PCR-HRM) assay. A significant positive correlation was observed between the SNP rs3077, rs9277535 of HLA-DP and SLE susceptibility (rs3077, OR = 0.74, 95%CI = 0.60–0.91, P = 0.004; rs9277535, OR = 0.72, 95%CI = 0.59–0.88, P = 0.001). Rs3077 polymorphism was corelated to IL-17, INF-γ and cutaneous vasculitis (P = 0.037, P = 0.020 and P = 0.006, respectively). Additionally, rs3077 AA genotype carriers showed lower concentration of inflammatory cytokines and lower cutaneous vasculitis incidence than did the other two genotype. No significant association was observed between rs9277535 and cytokines or any clinical features. In conclusion, HLA-DP polymorphisms (rs3077 and rs9277535) were associated with SLE susceptibility and the levels of some inflammatory cytokines in SLE patients.

目的:探索不同模式的牵张应力条件促进人类膀胱平滑肌细胞(human bladder smooth muscle cells,HBSMCs)增殖能力的生物学效应,以及这一过程背后的分子生物学机制.方法:将HBSMCs接种到硅胶膜上,对其施加单轴拉伸应力,其应变参数分别为2.5％,5％,10％和15％,牵张频率参数分别为0.05,0.10,0.20,0.50,1.00 Hz,并设置无应力施加的对照组进行研究.采用溴-脱氧尿嘧啶(bromodeoxyuridine,BrdU)掺入法量化分析细胞的增殖能力,摸索最适合HBSMCs增殖的牵张模式.采用实时PCR(real-time PCR,RT-PCR)技术和Western blotting技术分别从分子水平和蛋白质水平检测生理周期性牵张应力条件下HBSMCs的PI3K、SGK1、Akt、Kv1.3分子表达情况,以进一步研究应力促进细胞增殖的分子机制.结果:最适合HBSMCs增殖的牵张应力参数为应变5％、牵张频率0.10 Hz,因该试验组的细胞增殖参数较其他各试验组和对照组更高,其结果具有统计学意义(P<0.05).与对照组相比,牵张试验组的PI3K、SGK1、Kv1.3表达量显著升高(P<0.05),而Akt则无明显改变(P>0.05).分别应用SGK1和Kv1.3的阻滞剂重复细胞牵张试验均可消除上调的增殖能力.结论:周期性牵张应力可以促进HBSMCs增殖,其力学分子生物学信号可能通过PI3K-SGK1-Kv1.3通路转导,而PI3K-Akt通路并未参与其中.

OBJECTIVE:To explore the roles of bone metabolism-related molecules in bone metabolic disturbance and disease progression of patients with rheumatoid arthritis (RA).METHODS:A total of 66 RA patients and 20 healthy controls were included, and their relevant clinical information was gathered. Electrochemiluminescence immunoassay (ECLIA) was used to detect the levels of osteocalcin N-terminal middle (OC-N-MID) and C-terminal cross-linked telopeptides of type 1 collagen (CTX) in serum. The serum levels of Wnt inhibitory factor Dickkopf1 (DKK1) and receptor activator of nuclear factor-κB ligand (RANKL) were determined by magnetic luminex assays. Flow cytometry was used to detect the RANKL level on peripheral blood T cells.RESULTS:Compared with healthy controls, the levels of OC-N-MID and CTX in the sera of the RA patients showed no significant difference. Level of RANKL in the sera of the RA patients was higher than that in the controls, while level of DKK1 was lower. The level of RANKL on peripheral blood T cells increased in the RA patients, especially on CD3(+) T cells.CONCLUSION:In RA patients, level of RANKL on T cells is elevated, and the serum level of RANKL increases, but DKK1 decreases.

Objective E26 transformation specific sequence 1 (ETS-1) belongs to the ETS family of transcription factors that regulate the expression of various immune-related genes. Increasing evidence indicates that ETS-1 could contribute to the pathogenesis of autoimmune disease. Recent research has provided evidence that ETS-1 might correlate with rheumatoid arthritis (RA), but it's not clearly defined. In this study, we aimed to identify whether polymorphisms of ETS-1 play a role in Rheumatoid arthritis (RA) susceptibility and development in Chinese Han population. Methods Four single nucleotide polymorphisms (SNPs) within ETS-1 were selected based on HapMap data and previous associated studies. Whole blood and serum samples were obtained from 158 patients with RA and 192 healthy subjects. Genotyping was performed with polymerase chain reaction-high resolution melting (PCR-HRM) assay and the data was analyzed using SPSS17.0. Results A significantly positive correlation was observed between the SNP rs73013527 of ETS-1 and RA susceptibility, DAS28 and CRP (P<0.001, P = 0.001, and P = 0.028, respectively). Carriers of the haplotype CCT or TCT for rs4937333, rs11221332 and rs73013527 were associated with decreased risk of RA as compared to controls. No statistical significant difference was observed in the distribution of rs10893872, rs4937333 and rs11221332 genotypes between RA patients and controls. Conclusions Our data further supports that ETS-1 has a relevant role in the pathogenesis and development of RA. Allele T of rs73013527 plays a protective role in occurrence of RA but a risk factor in the high disease activity. Rs10893872, rs11221332 and rs4937333 are not associated with RA susceptibility and clinical features.

MicroRNAs (miRs) have crucial functions in spermatogenesis and implications for male infertility. In the present study, Homo sapiens (hsa)‑miR‑145 was designed and cloned into the eukaryotic expression plasmid pGenesil‑1. The recombinant plasmids were transfected into Hs 1.tes normal testicular cells and NTERA‑2 testicular cancer cells. Quantitative polymerase chain reaction of hsa‑miR‑145 indicated that pGenesil‑1‑miR‑145 effectively upregulated the expression of hsa‑miR‑145 in vitro. hsa‑miR‑145 overexpression inhibited the mRNA and protein expression of sex-determining region Y Box 9 in Hs 1.tes cells. The proliferation rates of NTERA‑2 cells transfected with pGenesil‑1‑miR‑145 were significantly decreased. High expression levels of miR‑145 promoted cell apoptosis in NTERA‑2 cells. The results revealed that altered hsa‑miR‑145 expression in testicular cells affects the regulation of target genes associated with male infertility.

OBJECTIVE:To assess the association of single nucleotide polymorphisms (SNPs) in exons of H2BFWT gene with male infertility in southwest China.METHODS:Three hundred and twelve infertile men and 211 fertile men were recruited. PCR was employed to amplify the target fragments of H2BFWT, and PCR products were sequenced. Prevalence of SNPs in the two groups was analyzed by statistical method.RESULTS:The detected SNPs have mainly distributed in the first exon of the H2BFWT gene. The ratios of 368G/A (rs553509) and -9C/T (rs7885967) were significantly higher in infertile group than fertile group. Additionally, a context-dependent effect was observed between 368G/A and -9C/T which the allele 368G combined with allele -9T will considerably increase the risk of male infertility.CONCLUSION:The present study has revealed that the SNPs in H2BFWT are associated with male infertility, and may increase the susceptibility of male infertility in southwest China.

The long-term benefits of antiviral treatment are limited by the resistance of hepatitis B virus (HBV).However, the effect of interferon (IFN)α treatment on drug-resistant HBVs is so far unknown.We, therefore, investigated the effects of IFN-α inducer poly-IC on the replication of HBV mutants resistant to drugs such as lamivudine (LAM), adefovir dipivoxil (ADV) and entecavir (ETV) in mice.HBV DNA and HBV DNA intermediate (RI) were employed as markers of the virus replication and 2',5'-oligoadenylate synthase (OAS) mRNA as a marker of IFN-α/β induction.Poly-IC inhibited wtHBV replication and increased levels of OAS mRNA.Compared to the wt virus, the capacity of virus replication was reduced in most LAMr and ETVr mutants except those with mutations rtM(204V+L180M+V173L), and was similary in the ADVr mutants except rt(A121V+N236T).The virus replication was reduced after poly-IC treatment with LAMr and ADVr mutants similary to the wt virus.In contrast, ETVr mutants were resistant to the poly-IC treatment.In conclusion, the capacity of HBV replication and the sensitivity to IFN therapy are influenced by drug-resistant mutations.The IFN therapy may effectively inhibit HBV replication in particular in patients with LAMr or ADVr mutations but not in patients with ETVr mutations.

We present a case of Ewing’s sarcoma and primitive neuroectodermal tumour (PNET) of the prostate. A 29-year-old male presented with difficult defecation and anus distention; on magnetic resonance imaging scan of the pelvis, we found a prostate tumour. A transrectal ultrasound-guided needle biopsy confirmed the diagnosis. The patient underwent cystoprostatectomy and replacement ileocystoplasty and was followed by multi-agent chemotherapy. PNET/ Ewing’s sarcoma of the prostate is extremely rare. The prognosis is very poor, so we should pay enough attention to the differential diagnosis and treatment.

OBJECTIVE:To assess the association between glutathione-S-transferase gene polymorphisms GSTT1, GSTM1 and GSTP1 and onset of azoospermia. METHODS:Multi-PCR was used to detect GSTM1 and GSTT1 gene deletions. Polymorphisms of GSTP1 were determined with restriction fragment length polymorphism (RFLP) method in 236 azoospermia patients and 142 healthy fertile male controls. RESULTS:The frequency of M1 (-/-) and P1 (Ile/Val or Val/Val) genotype was 24.65% in the control group, which was significantly higher than that of the patient group (15.68%, P=0.031). Frequency of M1 (-/-), T1 (+/+) and P1 (Ile/Val or Val/Val) genotype was 12.68% in the control group, which was significantly higher than that of the patient group (5.51%, P=0.014). CONCLUSION:The M1(-/-) and P1(Ile/Val or Val/Val) genotype and the M1(-/-), T1(+/+) and P1 (Ile/Val or Val/Val) genotype are associated with reduced risk of azoospermia in ethnic Chinese Han population.

Idiopathic azoospermia and oligospermia are one of the most important reasons for male infertility. Abnormal karyotype and azoospermia factor (AZF) microdeletion are two widely acknowledged reasons, but the most causes remain unclear. Y chromosome, as the male-specific chromosome, is closely related to the development of male reproductive system. To understand better the etiology of idiopathic azoospermia and oligospermia, we investigated the possible association between Y-haplogroup distributions and susceptibility to idiopathic azoospermia and severe oligospermia. Peripheral blood was collected from 193 men with normal reproductive history, 193 men with idiopathic azoospermia, and 72 men with idiopathic severe oligospermia. All the subjects underwent karyotyping and AZF deletion analysis to screen out those with AZF deletion and abnormal karyotype. The comparison of Y-haplogroup distribution between experimental group and control group was performed with SPSS V.18.0 software. Significant difference of Y-haplogroup distribution was observed in D1*, F*, K*, N1* and O3*(P=0.032, 0.022, 0.009, 0.009, 0.017, <0.05). The results suggest that Y chromosome haplogroup plays a important role in spermatogenic impairment.

We constructed a novel immunotoxin, hS83P34, by fusing the fragment containing the N-terminal 34 amino acids of human perforin to the C-terminal of humanized anti-CTLA-4 single chain fragment of variable antibody. In vitro cytotoxicity assays demonstrated that CTLA-4-positive activated human T cells and 6T-CEM were sensitive to hS83P34, while CTLA-4-negative resting T cells and endothelial cell ECV-304 were resistant to hS83P34. The IC50s of hS83P34 for activated T cells and 6T-CEM were about 0.2 μmol/L and 1.0 μmol/L, there was no obvious cytotoxicity of ECV-304 as detected at 8 μmol/L of hS83P34. In tumor graft rejection models, after treatment with 1.2 mg/kg immunotoxin every day for 12 days, the transplanted tumor cells were rescued by immunotoxin. The tumor weights of grafts of the rejection control group, nonrejection control group, and test group were 0.006 ± 0.014 g, 0.261 ± 0.048 g, and 0.135 ± 0.056 g, respectively. In the early 3 days posttransplantation, there were a lot of CD4- and CD8-positive T cells infiltrating into the tumor grafts of the rejection control group, while only a few T cells were detected in the tumor grafts of the test group. According to these results, we concluded that immunotoxin hS83P34 selectively depleted activated T cells in vitro as well as in vivo in an acute rejection model.