<Emphasis Type="Italic">FCGR3B</Emphasis> gene frequencies and<Emphasis Type="Italic"> FCGR3</Emphasis> variants in a Chinese population from Zhejiang Province

The human neutrophil antigens HNA-1a, -1b and -1c play an important role in immune neutropenia. The frequencies of the coding FCGR3B genes were determined in different populations. New FCGR3B variants were also found in some populations. This study investigated the FCGR3B gene frequencies and FCGR3 variants in a Chinese population compared with the results of Northern Germans and African Blacks (Uganda). Our results show that the gene frequencies in 413 healthy Chinese individuals from Zhejiang Province were 0.565 for FCGR3B*1, 0.430 for FCGR3B*2 and 0.00 for FCGR3B*3. The genotype frequency of FCGR3B_null was 0.48% (2/413). Sequencing of FCGR3 revealed that in seven out of 19 Chinese individuals, cloned and sequenced DNA fragments that exhibited variants caused by single nucleotide exchanges at one or more of the polymorphic positions 141, 147, 227, 266 and 277 in exon 3 also existed in this Chinese population. From the present study, it is concluded that the FCGR3B*1 gene is more frequent in a Chinese population from Zhejiang Province than the FCGR3B*2 gene, and the FCGR3B*3 gene seems to be absent, which is in contrast to studies in the white populations. Gene variants caused by single nucleotide exchanges were found in addition to the well-known forms, but the reason for this remains unclear.     

Linkage disequilibrium amongst ITGA2B and ITGB3 gene variants in patients with Glanzmann thrombasthenia confirms that most disease‐causing mutations are recent

We recently reported mutation analysis of the largest cohort of Glanzmann thrombasthenia (GT) patients so far examined. Sanger sequencing of coding regions, splice sites, upstream and downstream regions of the ITGA2B and ITGB3 genes identified 78 causal genetic variants (55 novel); 4 large deletions or duplications were also detected. We have now analysed the expression of non‐causal gene polymorphisms in the sequenced regions of both genes in selected members of this cohort. We identified 10 mostly silent variants in ITGA2B and 37 in ITGB3; all were present in control donor databases. Three non‐synonymous single nucleotide polymorphisms present were human platelet alloantigen (HPA) variants. A series of haplogroups, often including HPA‐3b in ITGA2B, repeated with little variation across unrelated families of wide geographical origins and with different GT‐causing mutations whether in ITGA2B or ITGB3. In contrast, a deleterious heterozygous c.1440‐13_c.1440‐1del in intron 14 of ITGA2B shared a common ITGA2B haplogroup composed of at least five gene polymorphisms and re‐occurred in seven European families with no known family relationships. Our results highlight the value of gene polymorphism analysis in GT and are consistent with the bulk of disease‐causing mutations in GT being of recent origin.     

Attainment of complete/very good partial response following rituximab-based therapy is an important determinant to progression-free survival, and is impacted by polymorphisms in FCGR3A in Waldenstrom macroglobulinaemia

The incorporation of rituximab into various regimens has improved depth of response in Waldenstrom macroglobulinaemia (WM), though the impact of achieving better responses remains to be determined. We examined response depth on progression‐free survival (PFS) in 159 rituximab‐naïve WM patients who received rituximab‐based therapy. The median follow‐up was 33·5 months, and categorical responses were as follows: complete response (CR, 8·8%); very good partial response (VGPR, 13·2%); partial response (50%); minor response (18·9%); Non‐Responders (8·8%). Sequencing for polymorphic variants of FCGR2A, FCGR2B, and FCGR3A was performed, and impact on response depth determined. Achievement of better categorical responses was incrementally associated with improved PFS (P < 0·0001). No separation was observed between CR and VGPR, and attainment of at least a VGPR was associated with improved time‐to‐progression. Neither age, serum IgM, haematocrit, platelet count, serum β₂microglobulin, WM International Prognostic Scoring System score, and treatment group predicted for CR/VGPR. Polymorphisms at FCGR3A‐48 and ‐158 were associated with improved categorical responses, particularly attainment of CR/VGPR (P ≤ 0·03). The attainment of CR/VGPR was associated with significantly longer PFS in rituximab‐naïve WM patients undergoing rituximab‐based therapy, and was predicted by polymorphisms in FCGR3A.     

重组柯萨奇B3病毒VP1基因的穿梭表达载体的构建及鉴定

目的　构建重组柯萨奇B3病毒VP1基因的细菌 -酵母菌穿梭表达载体pGBKT7-VP1。方法　用PCR从质粒pT7-CV3- 5 5中扩增柯萨奇B3病毒VP1基因 ,经NdeⅠ和PstⅠ双酶切后 ,定向插入细菌 -酵母菌穿梭表达载体 pG BKT7中完成质粒构建。用酶切和测序鉴定构建质粒。结果　限制性内切酶酶切和序列分析表明VP1巳成功插入pGBKT7载体的多克隆位点 ,克隆方向和阅读框与病毒VP1基因一致。结论　本研究成功构建VP1基因的细菌 -酵母菌穿梭表达载体 pGBKT7-VP1,为研究VP1蛋白与其宿主细胞之间的相互作用奠定了基础。     

柯萨奇病毒B3型VP1基因在酿酒酵母中的表达

目的在酿酒酵母AH109中表达柯萨奇病毒B3型VP1基因，并检测VP1蛋白对报告基因的转录自激活作用。方法用乙酸锂法将重组柯萨奇B3病毒VP1基因的细菌一酵母菌穿梭载体pGBKT7一VP1转化酵母菌。通过表型鉴定、PCR法验证质粒导入宿主菌后，Western blotting验证VP1蛋白的表达，最后经营养缺陷培养基和酶底物x—a-Gal反应检测VP1基因对报告基因的转录自激活作用。结果表型鉴定和PCR法均证实pGBKT7-VP1质粒转入酵母菌成功，Western blotting证实AH109能表达VP1蛋白，营养缺陷型培养基和酶底物X-a—Gal反应结果显示VP1蛋白对3种报告基因均无转录自激活作用。结论柯萨奇病毒B3型VP1蛋白可作为酵母双杂交系统的诱饵蛋白，为筛选与其相互作用的宿主细胞蛋白提供了基础。     

一例Rotor综合征SLCO1B1和SLCO1B3基因突变分析

目的初步探讨1例Rotor综合征患者的临床特点和SLCO1B1和SLCO1B3基因突变情况,从分子遗传学角度分析该疾病的发生机制。方法收集患者临床资料,从外周血白细胞提取基因组DNA,采用二代测序进行四千种已知单基因遗传性疾病筛查,用Sanger测序法分析验证二代测序发现的突变位点。结果患儿主要临床表现为反复皮肤及巩膜黄染,实验室检查示高胆红素血症,直接、间接胆红素双向增高。高通量测序发现患儿携带SLCO1B1基因c.1738 CT纯合突变和SLCO1B3基因c.360_481 del纯合突变。c.1738 CT突变为无义突变,已有文献报道,推测导致蛋白的第580位氨基酸密码子由精氨酸变为终止密码子。c.360_481 del突变为移码突变,蛋白编码区的第360至481位碱基缺失。该变异未见文献报道,也未见SNP数据库收录。此变异使蛋白缺失40个氨基酸的同时还造成开放阅读框移码,可能造成蛋白功能丧失。结论 SLCO1B1和SLCO1B3基因突变导致的有机阴离子转运多肽OATP1B1和OATP1B3功能缺陷是该Rotor综合征患者临床表现的分子遗传基础。     

柯萨奇病毒B组3型中国分离株VP1基因真核表达系统的克隆

目的探讨构建柯萨奇病毒Ｂ组３型（ＣＶＢ３）基因疫苗的可行性。方法应用逆转录ＰＣＲ技术扩增ＣＶＢ３中国分离株ＶＰ１基因,通过ＴＡ克隆法构建ｐＣＲ２．１－ＣＶＢ３ＶＰ１,将ＣＶＢ３ＶＰ１亚克隆至真核表达载体ｐＣＥＰ４,构建真核表达系统ｐＣＥＰ４－ＣＶＢ３ＶＰ１,并进行酶切和测序鉴定。结果发现ＣＶＢ３的中国分离株与Ｎａｎｃｙ株的ＶＰ１基因基本一致,均编码２９３个氨基酸,其中有５９处核苷酸存在变异,但仅改变了１０处氨基酸编码,证实克隆的片段为ＣＶＢ３ＶＰ１基因,表达载体为ｐＣＥＰ４。结论实验成功地构建了ＣＶＢ３中国分离株的真核表达系统ｐＣＥＰ４－ＣＶＢ３ＶＰ１,为ＣＶＢ３基因疫苗的研究奠定了基础     

中国大陆人散发性先天性巨结肠症患者内皮素受体B基因的研究

目的 探讨中国大陆人散发性先天性巨结肠症（sHD）易患基因内皮素受体B（EDNRB）的突变与多态性特征。方法 以92例sHD及其中32例患儿双亲为研究对象,并以60例正常儿为对照。提取受检者外周静脉血DNA,采用聚合酶链反应-单链构象多态性技术（PCR-SSCP）对EDNRB基因外显子-2（exon-2）进行分析,并通过DNA测序检测阳性标本的核苷酸改变方式,与文献报道的其他种族sHD同一基因特征做比较。结果 全部标本EDNRB基因exon-2均未发现突变与多态性位点的存在。结论 中国大陆人sHD患者EDNRB基因的exon-2不存在突变与多态性位点。     

Association of Ephrin receptor A3 gene polymorphism with susceptibility to chronic severe hepatitis B

Aim: Previous research has suggested that Ephrin receptor A3 (EphA3) plays signaling roles in the processes of inflammation by regulating lymphocyte migration and proliferation. In this study, we investigated whether the EphA3 gene polymorphism was associated with disease progression of chronic hepatitis B virus (HBV) infection. Methods: The EphA3 variant rs9310117 was genotyped in 1245 unrelated Han Chinese HBV carriers including 800 cases and 445 controls. χ² test was used to examine the difference in allele frequencies and genotype distributions between groups. The association between the polymorphism and disease progression of HBV infection was conducted by unconditional logistic regression analysis. Results: Statistical analysis revealed that the genetic variant was significantly associated with the occurrence of chronic severe hepatitis B (CSHB). We observed that subjects bearing at least one T allele (C/T or T/T genotype) had a decreased susceptibility to chronic severe hepatitis B compared with those bearing C/C genotype (P = 0.003, odds ratio = 0.560; 95% confidence interval, 0.381–0.824, recessive model). Genotype C/T had also been confirmed to protect subjects from suffering chronic severe hepatitis B (P = 0.001, odds ratio = 0.498; 95% confidence interval, 0.330–0.752, additive model). Conclusion: Our results suggest that the genetic alteration at EphA3 locus plays a role in the occurrence of chronic severe hepatitis B.     

ubiquilin3基因在小鼠组织中的表达及意义

目的 探讨ubiquilin3基因在小鼠心、肝、脾、肺、肾、脑、胃、小肠、大肠、睾丸、卵巢、子宫等多种组织及不同周龄小鼠睾丸组织的表达及其意义.方法 分别利用PCR、Western、Northern等方法,观察ubiquilin3基因在小鼠多种组织、不同周龄的小鼠睾丸组织,mRNA和蛋白水平的表达.结果 在mRNA和蛋白水平上,ubiquilin3基因在其他组织中均没有表达,而在小鼠睾丸组织中特异性表达;在不同周龄的小鼠睾丸组织中,从2周龄的小鼠睾丸组织开始表达,3周表达量逐渐增加,至4周小鼠中表达量达最高,且在成熟小鼠中持续表达.结论 ubiquilin3基因在小鼠睾丸组织特异表达,在成熟小鼠中表达量达到最高值,推测其与小鼠精子生成及精子成熟过程相关.     

高脂血症患者载脂蛋白B基因3''''VNTR大等位基因与血脂水平关系的研究

目的：本文旨在研究高脂血症患者载脂蛋白B基因3’VNTR讯大等位基因的频率分布和内部结构，及其多态性与血脂水平的相关性。方法：随机抽取2001—2002年间北京地区汉族人群高脂血症患者300例，正常对照300例，从高脂血症患者中选取携带apoB基因3’VNTR大等位基因重复数≥40（HVE≥40）的患者44例为高脂血症组，从正常对照中选取携带apoB基因3＇VNTR大等位基因重复数≥40（HVE≥40）的血脂正常对照33例为对照组，共77例。采用PCR、琼脂糖凝胶电泳等方法分析apo-B3＇VNTR大等位基因（HVE≥40）多态性，并测序分析其中部分等位基因的内部序列结构。结果：（1）高脂血症组apoB 3’VN’TR等位基因HVE50频率最高（23．5％），而对照组以HVE42频率最多（23．8％）。（2）血甘油三酯、血apoB、LDL水平越高，其3’VNTR等位基因长度越大的趋势，而与血TC无相关性。结论：（1）apoB基因3’VNTR大等位基因／大基因型与血甘油三酯、总胆固醇、血apoB增高相关联，提示3’VNTR大等位基因／大基因型可能是高脂血症的易感基因标志物。（2）apoB基因3’VNTR多态性可能是影响血脂水平差异的遗传因素之一。     

肾上腺素β3受体基因W64R多态性与苯那普利疗效之间的关系

目的探讨肾上腺素β3受体基因W64R多态性与原发性高血压患者苯那普利降压疗效之间是否存在相关性.方法在安徽省安庆及霍邱地区人群中筛选原发性高血压患者1170名,所有患者予苯那普利10 mg每日一次口服15 d,测定其用药前后的血压,并进行肾上腺素β3受体基因W64R基因型测定,观察其血压下降情况与基因型的关系.结果(1)在中国南方汉族原发性高血压患者中,ADRβ3W64R多态性位点的基因型频率是:WW基因型72.8%、WR基因型25.4%、RR基因型1.8%.(2)苯那普利10 mg每日一次口服15 d,女性患者收缩压下降水平在ADRβ3 W64R多态性位点各基因型之间差异具显著性,WR/RR基因型者比WW基因型者收缩压下降少3.49 mmHg(P=0.03).(3)药物治疗15 d后女性患者第16天收缩压降至140 mmHg以下、舒张压降至90 mmHg以下或下降幅度≥15%的达标率,WR/RR基因型低于WW基因型(OR=0.65,P＜0.05).结论在女性原发性高血压患者肾上腺素β3受体基因W64R多态性与原发性高血压患者苯那普利降压疗效之间存在相关性,可能为预测苯那普利降压疗效的一个预报因子,指导临床用药.     

Hepatobiliary phospholipid transporter ABCB4, MDR3 gene variants in a large cohort of Italian women with intrahepatic cholestasis of pregnancy

BACKGROUND: Intrahepatic cholestasis of pregnancy is a multifactorial disorder of pregnancy associated with a genetic background. AIM: To evaluate the genetic contribution of ABCB4, MDR3 gene in the development of intrahepatic cholestasis of pregnancy in a large cohort of Italian subjects. METHODS: This study represents an extension of a previous multicentre-prospective study including three Italian referral centres. In all, we enrolled 96 women at the 3rd trimester of pregnancy. Genomic DNA was extracted from peripheral venous blood leucocytes by standard procedures. Polymerase chain reaction was used to amplify exon 14, 15 and 16 of MDR3 gene. RESULTS: We found 3 non-synonymous heterozygous mutations in exon 14 (E528D, R549H, G536A), 1 in exon 15 (R590Q) and 2 in exon 16 (R652G, T6671). MDR3 gene variants in exons 14, 15 and 16 occurred in 7/96 of pregnant mothers with intrahepatic cholestasis of pregnancy (7.2%), and in none of 96 pregnant controls matched for age and parity. All seven patients had normal gamma-glutamyl transpeptidase, normal bilirubin, but high levels of both alanine transferase and serum bile acids. One had cholesterol biliary lithiasis. The outcome of pregnancy was normal in four cases (with vaginal delivery), while there was one fetal distress. CONCLUSIONS: MDR3 mutations are responsible for the development of intrahepatic cholestasis of pregnancy in only a small percentage of Italian women. Further genetic studies are warranted, however, to clarify the role of different mutations in intrahepatic cholestasis of pregnancy.     

STAT3基因在原发性肝癌组织中的表达及意义

目的探讨STAT3基因在肝癌发生发展中的作用及意义。方法应用RT-PCR检测STAT3基因及相关基因c-myc、p53、survivin、VEGF的mRNA水平的表达,应用Western Blot法及免疫组化法检测STAT3基因的蛋白水平及定位表达。结果肝癌组织及癌旁组织中STAT3基因mRNA的表达均明显高于正常肝组织(P<0.05),肝癌及癌旁组织中c-myc、survivin、VEGF基因mRNA的表达上调,p53基因mRNA的表达下调,肝癌组织及癌旁组织中STAT3基因蛋白水平的表达均高于正常肝组织(P<0.05)。结论STAT3基因的持续激活在肝癌的发生发展中起重要作用,可作为早期诊断的指标及治疗的新靶点。     

小RNA病毒3A基因及其编码蛋白研究进展

小RNA病毒3A蛋白是病毒的非结构蛋白,本文主要介绍3A基因以及其编码蛋白的特点,3A蛋白对细胞蛋白转运中的作用以及与其他蛋白的相互作用,3A蛋白与ARF家族在囊泡运输中作用的机制等.