Associations between IL-1RN variable number of tandem repeat, IL-1β (-511) and IL-1β (+3954) gene polymorphisms and urolithiasis in Uighur children of China

Objective:Interleukin-1(IL-1)is a pro-inflammatory cytokine which may be related to urolithiasis.Genetic polymorphisms of the interleukin-1beta(IL-1β)have been proposed as markers for urolithiasis in some areas.Due to the high incidence of urolithiasis in Uighur children(Xinjiang,China)and existence of ethnic difference,our aim is to explore the potential of IL-1 gene polymorphisms and urolithiasis among these children.Methods:Genomic DNA extracted from peripheral blood of 115 patients and 98 controls were used for genotype polymorphisms analyses.IL-1 receptor antagonist(IL-1RN)gene variable number of tandem repeat(VNTR)gene polymorphisms were analyzed by PCR method.PCR-based restriction analysis was done for the IL-1β(-511)and IL-1β(+3954)gene polymorphisms by endonucleases Ava I and Taq I,respectively.The genotype distribution,allele frequencies,carriage rate,and haplotype frequencies were statistically analyzed.Results:No significant differences were observed in genotypic frequencies between pediatric urolithiasis patients and control group for IL-1RN gene(χ^(2)=1.906,p=0.605),IL-1β(-511)gene(χ^(2)=0.105,p=0.949),or IL-1β(+3954)gene(χ^(2)=3.635,p=0.169).There were yet no significant differences of the allele frequencies of IL-1RN VNTR gene(p=0.779),IL-1β(-511)gene(p=0.941),and IL-1β(+3954)gene(p=0.418)in the case and control groups,as well as the carriage rate and haplotype of them(all p>0.05).Conclusions:The associations between IL-1RN VNTR,IL-1β(-511)and IL-1β(+3954)genes polymorphisms and urolithiasis were not significant in Uighur children.The results need to be confirmed in studies with larger population sample size,as well as in other ethnic groups.

Correlation between IL-1β,IL-1Ra gene polymorphism and occurrence of polycystic ovary syndrome infertility

Objective:To explore the relationship between IL-1β.IL-1Ra gene polymorphism and the occurrence of polycystic ovary syndrome(PCOS) infertility.Methods:A total of 59 PCOS infertility cases visiling the reproductive center of our hospital from Mar.2010 to Mar.2012 and 56 healthy women were selected.ELISA method was used lor the detection of IL-1β.IL-1Ra lewis,and the levels of serum supersensitivity C reaction protein(US-CRP).insulin(FINS),follieule-stimulating hormone(FSH) and fasting blood—glucose(FRG) were detected.PCR analysis technology was adopted to detect the gene polymorphism of the.511 site of IL-1βand the second introne of IL- 1Ra.Results:The levels of IL-1β.IL-1Ra.US-CRP.FINS and FBG in blood scrum of patients in PCOS group were significantly higher than those in control group(P【0.05 or P【0.01).The level of FSH in PCOS group was significantly lower than that in control group(P【0.05).The genotypic frequency of T/T.the 511 site of IL-1βin PCOS group was 42.37%.significantly higher than 1250%in control group 【P【0.01).The frequency of T allele was also significantly higher than that in control group(P【0.01).The genotypic frequency ofⅠ/Ⅴ.the second introne of IL-1Ra in PCOS group was 20.34%,signicianlly higher than 3.57%in control group(P【0.05).The frequency of V allele in PCOS group was significantly higher than that in control group(P【0.05).Conclusions: T allele of the 511 site of IL-1βgene and V allele of the second inlrone of IL-1Ra gene might be the genetic basis of the rising of IL-1β.IL-1Ra and US-CRP levels in blood serum of PCOS patients,and are associated with the infertility occurrence of PCOS patients.

Correlation between single nucleotide polymorphism of rs3811047 in IL-1 F7 gene and rheumatoid arthritis susceptibility among Han population in central plains of China

Objective:To discuss the association between single nucleotide polymorphism(SNP) of rs3811047 in IL-1 F7 gene and rheumatoid arthritis(RA) susceptibility among the Han population in central plains of China.Methods:A total of 276 RA patients admitted to our hospital from December 2009 to December 2011 together with 276 healthy physical examinees in the same period were chosen as the subjects.The typing for rs3811047 SNP in IL-1 F7 gene was carried out by using ligase detection reaction and polymerase chain reaction technique.And the frequency of each allele and genotypes distribution was calculated so as to evaluate the association between genotype distribution and RA susceptibility.Results:The frequency of A allele of rs3811047 in IL-1 F7 gene in RA group and control group was 16.27%and 17.68%,respectively,and that of G allele in two groups was 83.73%and 82.32%,respectively.The difference between two groups wasn’t statistical significant(P 】0.05).The frequency of genotype AA,AG and GG in RA group was 2.19%,27.84%and 69.97%,respectively,while that in control group was 2.94%,29.78%and 67.28%, respectively.The difference of distribution of three genotypes was not statistically significant (P 】0.05).RA patients with A allele were better than those without A allele in joint swelling index, rest pain,HAQ scoring and blood sedimentation.There was significant difference between two groups in above indexes(P【0.05/P【0.01).Conclusions:No significant correlation between RA susceptibility among the Han population in central plains of China and rs3811047 SNP inIL-1 F7 gene is observed.However,A allele of rs3811047 has certain influence on the condition of RA patients.

Genetic associations of inflammatory bowel disease in a South Asian population

AIM To estimate prevalence and phenotypic associations of selected inflammatory bowel disease(IBD)-associated genetic variants among Sri Lankan patients. METHODS A case study of histologically confirmed ulcerative colitis(UC) or Crohn's disease(CD) patients with ≥ 1 year disease duration, who were compared to unrelated, gender-matched, healthy individuals as controls, was conducted at four major centers in Sri Lanka. Phenotypic data of the cases were obtained and all participants were genotyped for 16 selected genetic variants: IL12 B :rs1045431, IL23 R :rs11805303, ARPC2 :rs12612347, IRGM :rs13361189, IL26/IL22 :rs1558744, CDH1 :rs1728785, IL10 :rs3024505, FCGR2 A :rs3737240, PTGER4 :rs4613763, IL17 REL/PIM3 :rs5771069, HNF4 a :rs6017342, STAT3 :rs744166, SMURF1 :rs7809799, LAMB1 :rs886774, HLA-DRB5, DQA1, DRB1, DRA :rs9268853, MST1, UBA7, and APEH :rs9822268. The genotypes of all variants were in Hardy-Weinberg Equilibrium(P > 10^(-3)). To account for multiple hypothesis testing, P-values < 0.003 were considered significant.RESULTS A total of 415 patients and 465 controls were recruited. Out of the single nucleotide polymorphisms(SNPs) tested, the majority were not associated with IBD in Sri Lankans. Significant positive associations were noted between rs886774(LAMB1-gene) and UC(odds ratio(OR) = 1.42, P = 0.001). UC patients with rs886774 had mild disease(OR = 1.66, P < 0.001) and remained in remission(OR = 1.48, P < 0.001). A positive association was noted between rs10045431(IL 12 B gene) and upper gastrointestinal involvement in CD(OR = 4.76, P = 0.002). CONCLUSION This confirms the heterogeneity of allelic mutations in South Asians compared to Caucasians. Most SNPs and disease associations reported here have not been described in South Asians.

Gene polymorphism in IL-1 receptor antagonist affects its production by monocytes in IgA nephropathy and Henoch-Schonlein nephritis

Objective To define the functional significance of IL-1 receptor antagonist (IL-1ra) gene polymorphism and to investigate, the production of IL-1ra by monocytes from individuals with different genotypes of IL-1.Methods The genotype of IL-1ra was detected by polymerase chain reaction (PCR). Peripheral monocytes obtained from patients with immunoglobin A nephropathy (IgAN), Henoch-Schonlein purpura nephritis (HSPN) and normal subjects were matched in sex and age between the IL1RN-2 allele carriers and non-carriers. The secretion of IL-1ra, IL-1α and IL-1β in the supernatant of GM-CSF (10ng/ml) treated and untreated monocytes were measured by ELISA.Results The secretion of IL-1ra by monocytes stimulated with GM-CSF was significantly higher in the IL1RN-2 allele non-carriers than those of carriers both in IgAN (21.55±3.08 vs 13.85±2.24ng/ml, P<0.001) and HSPN (23.72±6.68 vs 12.67±2.24ng/ml, P<0.01) as well as in normal controls (20.29±1.45 vs 10.51±2.3ng/ml, P<0.001). All showed no significant differences in monocyte secretion of IL-1α and IL-1β by GM-CSF stimulation between the IL1RN-2 allele carriers and non-carriers. Conclusions These results indicate that a functional correlation of the IL1RN-2 allele and IL-1ra production is present in patients with IgAN and HSPN. This gene polymorphism control of IL-1ra production may contribute to the variety of clinical responses to inflammatory stimulation in individuals with different genotype of IL-1ra.

类风湿关节炎与IL-1RAP基因多态性关联的研究

目的探索IL-1RAP基因多态性与类风湿关节炎(RA)的关联性。方法选取178例RA患者(RA组)及207例正常对照的全血标本,提取DNA、基因分型,对病例组与对照组IL-1RAP rs766442的等位基因频率、基因型进行分析。结果 IL-1RAP rs766442基因型包括GG、GT、TT,RA组与正常对照组间三种基因型频率差异有统计学意义(P=0.038)、等位基因频率差异有统计学意义(P=0.018)。结论携带T等位基因的RA患者出现RF-Ig G阳性的几率可能较高,IL-1RAP rs766442的TT基因型可能为RA的易感基因。

Effect of HIV-1 Tat on Secretion of TNF-α and IL-1β by U87 Cells in AIDS Patients with or without AIDS Dementia Complex

Objective To explore the role of HIV-1 tat gene variations in AIDS dementia complex （ADC） pathogenesis. Methods HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA. Results HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area （38-47aa）. All Tat proteins could induce ug7 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen. The level of Tat proteins derived from the ADC patient＇s spleen, basal ganglia, and the non-ADC patient＇s spleen were obviously higher than that from the non-ADC patient＇s basal ganglia. Conclusion Tat protein core functional area （38-47aa） may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.

miR-6867及其宿主基因RAPGEFL1在食管鳞状细胞癌组织和细胞系中的表达及甲基化状态

目的:检测miR-6867及其宿主基因Rap型鸟苷酸交换因子1(rap guanine nucleotide exchange factor like 1,RAPGEFL1)在食管癌细胞系及食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)组织中的表达水平及甲基化状态,并探讨其在ESCC发生发展中的作用。方法:选取河北医科大学第四医院2014年1月至2016年1月收治的ESCC手术患者组织标本87例。应用实时荧光定量PCR(qRT-PCR)检测miR-6867及RAPGEFL1在食管癌细胞系和ESCC组织及其相应癌旁组织中的表达水平,分析miR-6867和RAPGEFL1基因表达之间的相关性。应用甲基化特异性PCR法(MSP)检测RAPGEFL1在食管癌细胞系和ESCC组织及其相应癌旁组织中的甲基化状态。结果:miR-6867在ESCC组织中的相对表达量显著低于其相应癌旁组织(P<0.05),并与淋巴结转移及TNM分期有关(P<0.05);RAPGEFL1基因在ESCC组织中的表达显著低于其相应癌旁组织(P<0.05),并与淋巴结转移、组织分化程度及TNM分期相关(P<0.05);miR-6867与RAPGEFL1在ESCC组织中的表达呈明显正相关(P<0.05)。5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycytidine,5-Aza-dC)处理后,4种食管癌细胞系中miR-6867和RAPGEFL1基因的表达均增高,并且其甲基化程度明显降低。RAPGEFL1基因在ESCC组织中的甲基化率显著高于其相应癌旁组织(P<0.05),并与淋巴结转移、组织分化程度及TNM分期有关(P<0.05)。结论:ESCC的发生发展可能与miR-6867和RAPGEFL1的异常低表达及RAPGEFL1高甲基化状态有关,miR-6867与RAPGEFL1表达具有一致性,且RAPGEFL1基因启动子区甲基化可能是导致miR-6867与RAPGEFL1表达沉默的机制之一。

GAS1、IL-1RAP、PRF1在ALK阳性间变性大细胞淋巴瘤患者中的表达及其临床意义

【摘要】目的研究生长停滞特异性蛋白（GASl）、IL-1受体辅助蛋白（IL．1RAP）、穿孔素（PRFl）在间变性淋巴瘤激酶阳性的间变性大细胞淋巴瘤（ALK＋-ALCL）患者中的表达及其临床意义。方法以2011年1月至2016年9月的26例ALK-ALCL患者为研究对象，以12例ALK—ALCL、13例外周T细胞淋巴瘤非特指型和8例血管免疫母细胞性T细胞淋巴瘤患者为对照。收集所有患者的病理组织标本，采用实时荧光定量PCR和免疫组化法检测标本中GAS1、IL-1RAP、PRF1基因和蛋白表达水平，结合患者的临床资料对数据进行分析。结果①26例ALK＋ALCL患者组织中，GASl、IL-1RAP、PRFl基因和蛋白表达水平均高于3个对照组，差异有统计学意义（P值均〈0．05），对照组组间基因与蛋白表达水平差异无统计学意义（P〈0．05）。②存在LDH升高（0．77对1．38，z=-3．292，P=0．001）、国际预后指数（IPI）评分／〉3分（0．62对1．29，Z=-2．495，P=0．013）时，ALK＋ALCL患者的GASl基因表达水平降低；疾病分期为Ⅲ／Ⅳ期（0．89对1．18，z=-2．212，P=0．027）、IPI评分≥3分（0．48对1．13，Z=-2．008，P=0．045）时，ALK＋ALCL患者的PRFl基因表达水平降低；不同临床特征患者组间IL-1RAP基因表达水平差异均无统计学意义（P值均i〉0．05）。③化疗后达完全缓解的AL＋ALCL患者GASl、PRFl基因表达水平差异均无统计学意义（P值分别为0．016、0．009）。④以ALl（LALCL患者中各基因表达的中位数为分界点，GAS1、PRF1基因高表达组患者较低表达组有更长的总生存和无进展生存期（P值均〈0．05）。结论GASl、IL-1RAP、PRFl基因可作为ALK＋ALCL的分子标志，有潜在的诊断价值，可用于少数诊断困难病例的鉴别。GAS1、PRF1基因高表达的ALK＋ALCL患者疗效及预后更好。

IL-1β: an important cytokine associated with febrile seizures?

Febrile seizures （FSs） are the most common convulsions in childhood. Studies have demonstrated a significant relationship between a history of prolonged FSs during early childhood and temporal sclerosis, which is responsible for intractable mesial temporal lobe epilepsy. It has been shown that interleukin-1β （IL-1β） is intrinsically involved in the febrile response in children and in the generation of FSs. We summarize the gene polymorphisms, changes of IL-1β levels and the putative role of IL-1β in the generation of FSs. IL-1β could play a role either in enhancing or in reducing neural excitability. If the enhancing and reducing effects are balanced, an FS does not occur. When the enhancing effect plays the leading role, an FS is generated. A mild imbalance can cause simple FSs while a severe imbalance can cause complex FSs and febrile status epilepticus. Therefore, anti-IL-1β therapy may help to treat FSs.

蜱传牛巴贝斯虫套式PCR检测方法的建立及应用

为建立一种快速、敏感检测牛巴贝斯虫的方法,本研究根据GenBank中登录的牛巴贝斯虫rap-1基因保守区设计2对特异性引物,经反应条件的优化,建立了牛巴贝斯虫套式PCR检测方法。结果显示,该方法可以特异地检测牛巴贝斯虫,而对双芽巴贝斯虫、牛环形泰勒虫和弓形虫的检测均为阴性;该方法的灵敏度可达1.3×10~1拷贝/μL,是牛巴贝斯虫荧光定量PCR检测试剂盒的10倍,是常规PCR的1 000倍。对50只扇头蜱和微小牛蜱DNA进行检测,套式PCR、牛巴贝斯虫荧光定量PCR检测试剂盒和常规PCR的阳性检出率分别为100.0%、72.0%和0。本实验建立的套式PCR检测方法适用于牛巴贝斯虫病的早期诊断和分子流行病学调查,为蜱传牛巴贝斯虫病的防控提供技术支持。

双芽巴贝斯虫RAP-1C基因的克隆表达及重组蛋白反应原性的研究

拟通过对双芽巴贝斯虫棒状体相关蛋白1(rhoptry-associated protein-1,RAP-1)羧基端进行研究,以期了解该蛋白的一些免疫学特性。利用生物信息学软件,对RAP-1进行抗原表位预测,并与其他重要的巴贝斯虫和泰勒虫相应基因序列进行比对,以筛选出抗原性和特异性均良好的基因片段;针对筛选出的基因片段设计引物,对该基因进行克隆和原核表达,最后利用Western-blot对重组蛋白的反应原性和特异性进行分析。结果显示,融合目的蛋白含His标签,分子质量为26ku左右,该蛋白只与双芽巴贝斯虫阳性血清发生特异性反应,与其他主要巴贝斯虫、泰勒虫的阳性血清无交叉反应。结果表明,表达的PAP-1蛋白可作为ELISA诊断方法的候选抗原,为建立一种双芽巴贝斯虫病特异性的免疫学诊断方法奠定了基础。