宁夏回、汉族群体TGF-β1第1外显子区基因多态性分析

目的 探讨转化生长因子β1(TGF-β1)基因第1外显子区SNP+ 869 (T/C)和SNP+ 915 (G/C)多态性在宁夏回族、汉族人群中的分布特征.方法 用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术对210例回族和308例汉族人群转化生长因子β1第1外显子区(+ 869位点、+915位点)两个多态性位点进行检测.结果 宁夏回族群体TGF-β1基因第1外显子区两个多态性位点各基因型频率及等位基因频率分别为:+869(TT:25.24％;CT:52.38％; CC:22.38％;T:51.43％;C:48.57％);+915(GG:98.10％; GC:1.90％; CC:0％;G:99.04％;C:0.96％);宁夏汉族群体TGF-β1基因第1外显子区两个多态性位点各基因型频率及等位基因频率分别为:+869(TT:30.52％;CT:48.05％;CC:21.43％;T:54.55％;C:45.45％);+915(GG:99.03％;GC:0.97％;CC:0％;G:99.51％;C:0.49％),两组间无差异.按性别分组,亦无差异.结论 TGF-β1基因第1外显子区+869、+915两个多态性位点在宁夏回、汉族人群及不同性别间无差异.     

转化生长因子β1基因869T/C、915G/C多态性与食管癌关系的研究

目的研究转化生长因子β1(TGF-β1)第1外显子+869T/C、+915G/C基因多态性与广西地区食管癌的关系。方法采用序列特异性引物聚合酶链反应(PCR-SSP)技术,检测118例食管癌患者和130例正常对照组TGF-β1的基因多态性,同时采用酶联免疫吸附试验(ELISA)检测血清TGF-β1水平。结果食管癌患者血清TGF-β1水平显著高于对照组(P<0.01),TGF-β1基因+915G/C多态性各等位基因及基因型频率在两组人群中的分布差异有统计学意义(P<0.05),等位基因频率的相对风险分析发现,C等位基因携带者患食管癌的风险是G等位基因的3.077倍(OR=3.077,95%CI1.336~7.087),携带C等位基因食管癌患者血清TGF-β1水平显著高于不携带者[(55.37±9.76)μg/Lvs(48.29±8.29)μg/L,P<0.05];而TGF-β1基因+869T/C多态性在食管癌组和正常人群中的分布差异无统计学意义(P>0.05)。结论TGF-β1基因+915G/C多态性与食管癌的发病具有相关性,其中C等位基因可能是食管癌发病的遗传易感基因;携带C等位基因的个体可能通过促进TGF-β1的高度表达进而增加了食管癌的发病风险。     

新疆哈萨克族转化生长因子β1基因多态性及其血浆水平与原发性高血压的关系

目的 研究新疆哈萨克族原发性高血压(essential hypertension,EH)患者转化生长因子β1(transforming growth factor-β1,TGF-β1)+869T/C、+915G/C基因多态性及血浆水平与EH的关系.方法 采用聚合酶链反应-限制性片段长度多态性和基因测序对新疆哈萨克族365名EH患者及435名正常对照组进行基因分型,用双抗体夹心法测量TGF-β1血浆浓度.结果 +915G/C位点基因型GG、GC及等位基因G、C频率依次为97.9%、2.1%、98.77%、1.23%,EH组与对照组差异无统计学意义(P＞0.05);+869T/C位点基因型TT、TC、CC及等位基因T、C在对照组中频率依次为25.97%、46.67%、27.36%、49.3%、50.7%,CC基因型及C等位基因频率在EH组中高于对照组(41.60%vs.27.36%、62.2%vs.50.7%),差异有统计学意义(P＜0.05),C等位基因携带者EH患病风险高于T等位基因携带者(OR=1.6O,P=0.00).+869T/C与+915G/C存在连锁不平衡,其形成的单倍型C-G在EH组中频率高于对照组(61.6%vs.49.8%,P＜0.05).+869T/C及+915G/C基因型、等位基因在EH组和对照组中TGF-β1血浆水平差异无统计学意义(P＞0.05).结论 新疆哈萨克族TGFβ1+915G/C基因变异频率很低,且不存在纯合变异,+869位点C等位基因可能是哈萨克族EH的遗传易感基因,+869T/C与+915G/C多态性位点存在连锁不平衡,两者构成的单倍型C-G是EH危险性因素.     

白细胞介素13基因多态性与慢性阻塞性肺病肺功能的关系

目的研究白细胞介素13(IL-13)基因多态性与慢性阻塞性肺病(COPD)患者肺功能的关系。方法采用聚合酶链反应--限制性酶切片段长度多态性(PCR-RFLP)方法检测92例COPD患者IL-13基因+2044位点单核苷酸多态性,测定COPD患者肺功能,分析IL-13基因+2044位点多态性与肺功能的关系。结果 IL-13基因+2044位点基因型频率为G/G型48.9%、G/A型45.7%、A/A型5.4%,G/G型患者肺功能FEV1和FEV1/FVC(%)明显低于G/A型和A/A型(P<0.05)。结论 IL-13基因+2044位点多态性与COPD患者肺功能有关,G/G型COPD患者肺功能损害更加严重。     

TGF-β1基因启动子-800G/A、-509C/T多态性与食管癌的研究

目的研究转化生长因子β1(TGF-β1)基因启动子多态性各等位基因及基因型在食管癌患者中的分布频率,初步分析其基因型及血清水平与食管癌的相关性.方法采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,检测118例食管癌患者和130例正常对照组TGF-β1的基因多态性,包括TGF-β1基因启动子-800G/A、-509C/T位点,同时采用ELISA检测血清TGF-β1水平.结果食管癌患者血清TGF-β1水平显著高于对照组(P＜0.01),TGF-β1基因-800G/A位点多态性在食管癌组和正常人群中的分布差异无显著性(P＞0.05),而TGF-β1基因-509C/T多态性各等位基因及基因型频率在两组人群中的分布差异存在显著性(P＜0.05);等位基因频率的相对风险分析发现,T等位基因携带者患食管癌的风险是C等位基因的1.624倍(OR=1.624,95%CI1.134～2.324),携带T等位基因的食管癌患者血清TGF-β1水平显著高于不携带者(50.97±8.91μg/LVS44.23±8.54μg/L,P＜0.01).结论TGF-β1基因-509C/T多态性与食管癌的发病具有相关性,其中T等位基因可能是食管癌发病的遗传易感基因;携带T等位基因的个体可能通过促进TGF-β1的高度表达进而增加了食管癌的发病风险.     

三种基因SNP与BAVM易感及出血风险的相关性研究

目的探讨白细胞介素-17（IL-17A）、转化生长因子-β（TGF-β）及其受体（TGFRl32）的单核苷酸多态性（SNP）与脑动静脉畸形（BAVM）易感及出血风险的相关性。方法前瞻性收集BAVM患者外周血（n=53）,健康对照人群来自体检中心（n=120）。采用聚合酶链反应．限制性片段长度多态性（PCR—RFLP）法,检测IL-17A-197G／A,TGF-β1—509C／T及TGFR-β2—875A／G基因的SNP特征,并做关联分析探讨以上基因SNP与BAVM易感及出血风险的相关性。结果BAVM组与对照组比较,IL-17A-197G／A和TGF-β1．509C／T基因型及基因频率分布上的差异无统计学意义（P〉O．05）,TGFR-β2—875A／G基因型及基因频率分布差异有统计学意义（P〈0．05）;BAVM出血组IL-17A-197G／A的G／G基因型和TGFR-β2—875A／G的G基因频率明显高于未出血组（P〈O．05）。结论TGFR-β2．875A／G的G／G基因型可能是中国南方人群易感BAVM的危险因素．IL-17A-197G／A的G／G基因型可能与BAVM易破裂出血风险有关。     

云南汉族人群IL-4基因5’和3’非编码区域多态性与HCV慢性感染无相关性

目的探讨IL-4基因5’和3’非编码区域单核苷酸多态性位点(single nucleotide polymorphism,SNP)与HCV慢性感染的相关性。方法选取云南地区汉族人群HCV慢性感染患者380例,健康体检人群439例。采用Taq Man探针基因分型方法对IL-4基因5’和3’非编码区域6个SNP位点SNP-1138A/G(rs2243247)、-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)、3’端C/T(rs2243292)进行基因分型,并构建单倍型,评估上述6个SNP位点及单倍型与HCV慢性感染的相关性。结果 IL-4基因SNP-1138A/G(rs2243247),3’端C/T(rs2243292)在病例组和对照组中无多态性;SNP位点-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)的基因型频率和等位基因频率在病例组和对照组中差异无统计学意义(P0.05);单倍型分析结果显示:SNP位点-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)构建的单倍型频率在病例组和对照组中差异无统计学意义(P0.05)。结论在云南汉族群体中,IL-4基因5’和3’非编码区域SNP位点-1138A/G(rs2243247)、-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)、3’端C/T(rs2243292)与HCV慢性感染没有相关性。     

IL-10RA基因多态性与SLE发病关系的临床研究

目的:研究中国北方人群白细胞介素10受体A(IL-10RA)基因多态性分布情况,探讨IL-10RA基因变异与系统性红斑狼疮发病的相关性.方法:利用基因测序、PCR扩增及单链构象多态性(Single-strand Conformation Polymorphism,SSCP)电泳技术,对IL-10RA基因多态性进行筛选及分析,比较系统性红斑狼疮病例组与健康对照组中基因型的分布频率.结果:在IL-10RA第5内含子处存在3种有意义单核苷酸多态性(SNP),即T/T型、C/C型、C/T型.其中T/T型和C/T型在病例组中的出现频率明显增加.结论:IL-10RA内含子5具有多态性并与系统性红斑狼疮的发病相关.     

广州汉族葡萄膜炎患者TGF-β1＋869T／C基因多态性

目的探讨转化生长因子（TGF）-β1＋869T／C基因多态性与葡萄膜炎病的关系。方法运用多聚酶链反应技术检测广州汉族75例葡萄膜炎及140例正常人TGF-β1＋869T／C基因多态性的分布。结果葡萄膜炎组TGF-β1＋869T／C基因多态性基因型频率（％）分别为38．67、48．00、13．00,T、C基因频率分别为0．6267和0．3733,而对照组TGF-β1＋869T／C基因型频率（％）分别为38．57、48．57、12．86,对照组TGF-β1＋869C,T、C基因频率分别为0．6286和0．3714;TGF-β1＋869T／C基因多态性各种基因型频率在患者组与正常对照组之间的差异无统计学意义（xz=0．16,P〉0．05）。结论TGF[31．509C／T基因多态性与葡萄膜炎无明显相关。     

白细胞介素-10 基因启动子多态性与慢性阻塞性肺疾病易感性的关系

目的　探讨中国汉族人白细胞介素 - 10基因启动子单核苷酸多态性及其与慢性阻塞性肺疾病易感性之间的关系。　方法　应用聚合酶链反应 -限制性片段长度多态性分析方法 ,检测 94名健康吸烟者和 88例吸烟慢性阻塞性肺疾病 (chronic obstructive pulmonary disease,COPD)患者白细胞介素 - 10(interleukin- 10 ,IL- 10 )基因启动子 - 10 82 G/ A、- 819C/ T、- 5 92 C/ A单核苷酸多态性位点基因型。　结果共发现 11种启动子基因型 ,以 AA·TT·AA、AA·TC· AC、AA· TC· AA基因型多见 ;通过对 11种启动子基因型进行分析 ,新发现 ATC、ACA两种单倍型 ;健康吸烟者和吸烟 COPD患者 IL- 10基因启动子- 10 82 G/ A、- 5 92 C/ A位点基因型分布频率差异无显著性 ,- 819C/ T多态性位点与中国汉族人 COPD易感性有关 ;中国汉族人 IL- 10基因启动子等位基因频率与日本人相似 ,与白种人之间差异存在显著性。　结论　中国汉族人 COPD易感性与 IL- 10基因启动子 - 819C/ T位点多态性有关 ;中国汉族人 IL- 10基因启动子至少存在 ATA、ACC、GCC、ATC、ACA5种单倍型。     

Association of cytokine gene polymorphisms with the complications of allogeneic haematopoietic stem cell transplantation

The purpose of our study was to confirm the prevalence of the association between single nucleotide polymorphisms present in genes encoding cytokines and the complications occurring after haematopoietic stem cell transplantation (HSCT). 108 recipients and 81 donors were typed for TNF-α ( ?308), TGF-β1 (codon 10, 25), IL-10 (?1082, ?819, ?592), IL-6 (?174) and INF-γ (+874). Our studies have shown a tendency toward association between the occurrence of acute form of graft versus host disease (aGVHD) and IL-6 genotype. Homozygote C/C was less likely to develop aGVHD (p = 0,09). Genotype GCC/ATA in IL-10 recipient gene alone had protective effect against the occurrence of aGVHD (p = 0,01). Furthermore, GCC/ATA protected the host against developing the disease in the clinically relevant grades (II-IV) (p = 0,03). In addition, the recipient’s T/T G/G genotype (TGF-β1) predisposed to the development of both acute (p = 0,06 – trend) and chronic (p = 0,04) GVHD and also severe aGVHD (p = 0,004). We also observed a statistically significant association between the genotype of recipient and the risk of infection – the protective function of the G/C IL-6 in the bloodstream infections (p = 0,001). Our results suggest that IL-6, IL-10 and TGF-β1 genotypes of recipient are the most associated with the risk of complications after HSCT.     

-174G/C interleukin-6 gene polymorphism and the risk of transplanted kidney failure or graft loss during a 5-year follow-up period

The influence of cytokine gene polymorphisms on transplanted kidney outcome is not well understood. The aim of this one-centre study was to analyse the association between tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10, interferon-γ (IFN-γ) and transforming growth factor-β1 (TGF-β1) genotypes and the incidence of delayed graft function (DGF), acute rejection (AR) and 5-year kidney graft loss. Genotyping was performed in 199 subsequent kidney graft recipients from deceased donors without induction therapy based on polymerase chain reaction method using sequence-specific primers for TNF-α (-308A/G), IL-10 (-1082A/G, -819T/C and -592A/C), IL-6 (-174G/C), IFN-γ (+874T/A) and TGF-β1 (in codons 10T/C and 25G/C). Genotypes were grouped according to the strength of cytokine expression. During a 5-year follow-up period, 14 patients died with functioning graft and 33 developed graft failure. The analysed polymorphisms were not associated with the incidence of DGF. The frequency of early episodes of AR was significantly associated only with TGF-β1 genotype. There was an association between -174G/C IL-6 gene polymorphism and the death-censored kidney graft survival. The risk of graft loss during 5-year follow-up period was greater by 57% for GG or GC (higher IL-6 production) than for CC carriers. None of the other analysed polymorphisms significantly influenced both patients and kidney graft survival, also in the analysis of the subgroup with human leucocyte antigen-DR mismatch. -174G/C IL-6 genotype of the kidney graft recipient could modulate the rate of graft excretory function deterioration and the risk of graft loss by influencing their constitutional expression.     

The Expression of Th17-Associated Cytokines in Human Acute Graft-versus-Host Disease

The role of Th17 cells and Th17-associated cytokines in the development of acute graft-versus-host disease (aGVHD) in clinical allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients is not well established. In the current study, a cohort of 69 allo-HSCT patients was examined for the percentages of Th17 and FoxP3⁺ Treg cells and the expressions of RORγt and FoxP3 in peripheral blood mononuclear cells (PBMCs). The Th17 percentage and RORγt expression were significantly higher, whereas Treg percentage and FoxP3 expression were significantly lower in severe aGVHD (grade 3 to 4) and mild aGVHD (grade 1 to 2) patients than in patients without aGVHD (grade 0) and healthy donors. We then investigated the expressions of Th17-associated cytokines, including TGF-β, IL-6, IL-1β, IL-17, IL-21, IL-22, IL-23, as well as IL-23R in the PBMCs of patients after allo-HSCT. The expressions of IL-17 and IL-22 in CD4⁺ T cells were also examined. The results showed that the expressions of IL-6, IL-1β, IL-17, IL-21, IL-23, and IL-23R were all increased, whereas IL-22 expression was decreased in aGVHD patients. The changes were also correlated with the severity of aGVHD. We also investigated the dynamic changes of Th17/Treg cells and Th17-associated cytokines in patients during the onset and resolution of aGVHD. The results demonstrated a reciprocal relationship between Treg and Th17 cells. Th17-associated cytokine expressions, namely IL-17 and IL-23, were closely related to the occurrence and resolution of aGVHD. We conclude that the dynamic balance between the Th17 and FoxP3⁺ Treg cells and the changes of Th17-associated cytokines could be the indicators of the disease progression and promising candidates of prognostic biomarkers of aGVHD.     

Polymorphisms of transforming growth factor-β1 associated with increased risk of gastric cardia adenocarcinoma in north China

Transforming growth factor beta 1 (TGF-β1) is a multifunctional cytokine that has been implicated in the oncogenesis and tumour progression. However, the association of TGF-β1 polymorphism with gastric cardia adenocarcinoma (GCA) remains unclear. The aim of the study was to investigate the possible association of the polymorphisms of TGF-β1 with susceptibility to GCA in a population of north China. A case-control analysis was performed to assess the association of six single nucleotide polymorphisms (SNPs) of TGF-β1 and GCA risk. The genotype and allele distributions of TGF-β1 G-800A, C-988A, G915C and C788T in GCA patients were not significantly different from that in healthy controls (P>0.05). The -509T and 869C allele significantly elevated the risk of developing GCA (adjusted OR=1.45 and 1.41; 95% CI=1.04-2.10 and 1.07-2.08, respectively). The CT and TT genotype of C-509T and the TC and CC genotype of T869C significantly elevated the risk of developing GCA. When stratified by tumour stage, the -509T and 869C allele carriers had an increased risk of TNM stage III+IV GCA as compared with noncarriers. The C-509T and T869C SNP are in a strong linkage disequilibrium (D'=0.94). Compared with C/T haplotype, T/C haplotype significantly increased the risk of developing GCA. The TGF-β1 level and expression were higher in GCA patients with -509T or 869C allele than in those without T or C allele (P<0.05). GCA patients with -509TT and 869CC genotype had higher apoptotic tumour-infiltrating lymphocytes in their cancer tissues than those with -509CC and 869TT genotype. In all, TGF-β1 C-509T and T869C polymorphisms may be associated with an increased risk of GCA in north China.     

The effects of gene polymorphisms on susceptibility to acute GVHD and survival of allogeneic HSCT recipients: IL-10 gene polymorphisms as a more accessible target to predict prognosis

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a therapeutic modality commonly used to treat hematological and immunological disorders. Among the main complications of allo-HSCT is the acute graft-versus-host disease (a-GVHD), a condition which accounts for a high incidence of mortality. Several genes encoding inflammatory mediators may present polymorphisms, which have been implicated in the risk of developing a-GVHD. In our study, we investigated the association between genotypes of cytokine-encoding genes and the incidence and severity of a-GVHD and survival of HSCT recipients. No statistically significant association was found between IL and 6-174 G/C, INF-γ + 874 T/A, TNF-α -238 A/G, -308 A/G and IL-10-819C/T, -592 A/C polymorphisms and the presence or severity of a-GVHD. A higher risk of a-GVHD was associated with the IL-10-1082 GG genotype compared to the AA + AG genotypes of recipients and donors. The IL-10-1082 genotype can be used as a prognostic determinant to predict which HSCT recipient will be more responsive to the transplant. Thus, cytokine gene assays may be useful in the individualization of prophylactic regimens and for an appropriate selection of immunosuppressants based on the HSCT recipient’s responsiveness.     

Cytokine gene polymorphisms and graft-versus-host disease in children after matched sibling hematopoietic stem cell transplantation: a single-center experience

Various polymorphisms in cytokine genes have recently been investigated as candidate risk factors in allogeneic hematopoetic stem cell transplantation (allo-HSCT). We retrospectively analyzed specific polymorphisms in genes for interleukin (IL)-10, IL-6, tumor-necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) in a pediatric cohort of 57 histocompatibility leucocyte antigen (HLA)-identical sibling myeloablative transplants. Both recipient and donor genotypes were tested for association with graft-versus-host disease (GVHD) by statistical methods including Cox regression analysis. We found a significant association between the IL-10 promoter haplotype polymorphisms at positions -1082, -819 and -592 with the occurrence of severe (grades III-IV) acute GVHD (aGVHD). Recipients with the haplotype GCC had a statistically significant decreased risk of severe aGVHD (hazard risk (HR)=0.20, 95% confidence interval (CI): 0.06-0.67) in comparison with patients with other IL-10 haplotypes (P=0.008). Transplant-related mortality at 1 year was significantly lower in recipients with this haplotype (HR=0.17, 95% CI: 0.012-0.320) versus other IL-10 haplotypes (P=0.03), whereas overall survival was not influenced by IL-10 haplotype polymorphisms. In multivariate analysis, the presence of the IL-10 GCC haplotype was found as the only variable associated with a statistically significant decreased hazard of severe aGVHD development (P=0.02, HR=0.21, 95% CI: 0.05-0.78). These results suggest that pediatric patients possessing the IL-10 GCC haplotype may be protected from the occurrence of severe aGVHD in the setting of matched sibling HSCT.     

肿瘤坏死因子受体基因单核苷酸多态性与肺炎严重程度的相关性

 目的：比较肿瘤坏死因子受体（tumor necrosis factor receptor, TNFR）基因多个等位基因在肺炎人群中的分布频率,分析基因多态性与肺炎发病率和病情严重程度的相关性。方法：纳入66例肺炎患者与66例既往无肺炎的健康体检者,抽提各研究对象外周血DNA,通过聚合酶链式反应-限制性片段长度多态性或基因测序的方法检测TNFR1+36A/G、TNFR1-609G/T、TNFR2+676T/G、TNFR2 +1663T/G、TNFR2 +1668A/G和TNFR2 +1690C/T各多态性位点在肺炎患者与健康体检者、重症肺炎与非重症肺炎患者中的分布频率,并统计分析各基因分布频率与肺炎发生率和严重程度的相关性。结果：TNFR1-609G与T等位基因在肺炎患者中分布频率分别为40.9%与59.1%,在健康体检者中的分布频率分别为53.8%与46.2%; TNFR1-609T等位基因在肺炎患者中的分布频率较高（P<0.05）。余基因的各等位基因在肺炎患者与健康体检者中的分布频率差异无统计学意义。TNFR1-609G与T等位基因在重症肺炎患者中分布频率分别为25.0%与75.0%,在非重症肺炎患者中的分布频率分别为46.0%与54.0%,T等位基因在重症肺炎患者中的分布频率较高（P<0.05）。TNFR2 +1690C与T等位基因在重症肺炎患者中分布频率分别为81.1%与18.9%,在非重症肺炎患者中的分布频率分别为61.0%与39.0%,C等位基因在重症肺炎患者中分布频率较高（P<0.05）,余基因的各等位基因在重症肺炎与非重症肺炎患者中的分布频率差异无统计学意义。结论：携带TNFR1-609T等位基因的个体更易罹患肺炎,而携带TNFR1-609T及TNFR2 +1690C等位基因的个体肺炎病情较严重,易进展为重症肺炎。