简介:摘要目的对一个遗传性球形红细胞家系致病基因进行鉴定和突变致病性分析。方法采集该家系共17人的外周血样。采用高通量测序分析先证者外周血DNA,筛选候选致病变异并进行家系共分离分析。然后采用同源重组构建pCAS2c.5798+1G和pCAS2c.5798+1A型质粒,转染293T细胞,应用反转录PCR、TA克隆和Sanger测序分析候选致病变异对剪接的影响,同时提取患者外周血RNA,分析候选致病变异在体内的剪接情况。结果高通量测序结果显示先证者携带SPTB基因c.5798+1G>A变异,且该变异与家系患者的表型共分离。体、内外剪接实验结果显示c.5798+1G>A变异明显影响RNA的正常剪接,导致阅读框移码产生提前终止的密码子。结论SPTB基因c.5798+1G>A新变异是导致该家系遗传性球形红细胞症的原因。
简介:摘要目的对1个腓骨肌萎缩症(Charcot-Marie-Tooth,CMT)家系进行全基因组测序,为临床诊断和遗传咨询提供依据。方法应用高通量基因测序(next generation sequencing technology,NGS)检测CMT的致病基因位点,筛查各成员携带致病基因的情况。结果测序结果显示家系中两例患儿的GDAP1基因存在c.1A>G纯合变异,导致编码蛋白第1位氨基酸由Met变为Val(p.Met1Val),为错义变异,其父母均携带c.1A>G杂合变异;目前尚无该变异的相关报道,在人群中发生的频率极低。另外家系中1例患儿和母亲携带BAG3基因c.710A>T杂合变异。结论GDAP1基因c.1A>G纯合变异可能为该家系患儿的致病原因,基因测序结果为患儿的临床诊断和遗传咨询提供了依据。
简介:AbstractBackground:Epithelial to mesenchymal transition (EMT) is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma (HCC). Follistatin (FST) family members are considered to be an attractive therapeutic targets and prognostic indicators in cancers. As a derivative of FST, Follistatin Like 5 (FSTL5) may play a similar role in HCC cells. This study aimed to investigate the expression and function of FSTL5 in HCC and its role in EMT.Methods:FSTL5, E-cadherin and vimentin in HCC, and paracancerous tissues were detected by immunohistochemistry. Correlation of FSTL5 expression with overall survival was assessed. The proliferation and invasion of HCC cell lines SK-Hep1 and MHCC-LM3 were analyzed by cell counting kit-8 and Transwell assays. The expression of FSTL5, E-cadherin, and vimentin in HCC cells was examined by polymerase chain reaction and Western blot analysis. T-test was used to analyze the difference in proliferation and invasion ability between groups. The Spearman rank correlation test was used to detect the correlation between the expression of FSTL5 and E-cadherin or vimentin.Results:The expression of FSTL5 in HCC was lower than that in paracancerous tissues (9.97% vs. 82.55%, χ2 = 340.15, P < 0.001). Patients with high FSTL5 expression had a better prognosis (χ2= 8.22, P= 0.004) and smaller tumor diameter (χ2 = 45.52, P < 0.001), less lymph node metastasis (χ2 = 5.58, P= 0.02), earlier tumor node metastasis stage (χ2 = 11.29, P= 0.001), a reduced number of tumors (χ2 = 5.05, P= 0.02), lower alpha-fetoprotein value (χ2 = 24.36, P < 0.001), more probability of hepatitis carrying (χ2 = 40.9, P < 0.001), and better liver function grade (χ2 = 5.21, P = 0.02). Immunohistochemistry showed that FSTL5 expression in HCC tissues was positively correlated with E-cadherin expression (r = 0.38, P < 0.001) and negatively correlated with vimentin expression (r = -0.385, P < 0.001). Furthermore, over-expression of FSTL5 up-regulated the expression of E-cadherin and down-regulated the expression of vimentin in SK-Hep1 (negative control [NC] vs. FSTL5-interfering group [Lv-FSTL5]: E-cadherin [t= 45.03, P < 0.001], vimentin [t= 67, P < 0.001]) and MHCC-LM3 (NC vs. Lv-FSTL5: E-cadherin [t = 50, P < 0.001], vimentin [t = 72.75, P < 0.001]) cells at mRNA level. The same as protein level. In addition, the over-expression of FSTL5 inhibited the proliferation (NC vs. Lv-FSTL5: SK-Hep1, 3 d [t = 7.324, P = 0.018], 4 d [t = 6.23, P = 0.021], 5 d [t= 10.21, P= 0.003]; MHCC-LM3, 3 d [t= 4.32, P= 0.037], 4 d [t= 7.49, P= 0.012], 5 d [t= 9.3661, P = 0.009]) and invasion (NC vs. Lv-FSTL5: SK-Hep1, t= 21.57, P < 0.001; MHCC-LM3, t= 18.04, P < 0.001) of HCC cells.Conclusions:Down-regulation of FSTL5 may contribute to EMT of HCC, and FSTL5 is a potential target in the treatment of HCC.
简介:摘要目的分析1个近亲婚配的凝血因子Ⅴ缺乏症(coagulation factor V deficiency)家系的遗传学病因。方法应用高通量外显子测序筛查F5基因的变异,Sanger测序验证检出的变异,分析双亲携带变异位点的情况。结果先证者F5基因第13外显子存在c.4096delC纯合缺失,可导致移码变异,使FⅤ蛋白编码提前终止(p.Leu1366Phefs*3);先证者父母均为该变异的携带者。F5基因c.4096delC变异未见报道,根据ACMG指南推荐标准,为致病性变异。结论F5基因c.4096delC (p.Leu1366Phefs*3)纯合变异是该家系先证者发生凝血因子Ⅴ缺乏症的遗传学病因。新变异的检出丰富了F5基因的变异谱。
简介:摘要目的对多重实时定量PCR(real-time quantitative PCR,qPCR)进行反应体系筛选和方法探索,使其用于G1—G4型轮状病毒VP7基因的快速分型和定量分析。方法设计G1—G4型轮状病毒VP7基因特异性引物和探针,以特异性体外转录RNA为模板,筛选多重qPCR反应体系,建立多重qPCR方法。采用单因素方差分析和配对样本t检验对多重与单重qPCR检测结果进行比较。结果经筛选,得到6组三重qPCR反应体系和1组四重qPCR反应体系。所建立的三重和四重qPCR中,除四重qPCR的G1型参数〔标准曲线决定系数(R2)为0.982、扩增效率为89.221%〕略低于要求外,其余标准曲线R2均>0.99,扩增效率均在90%至110%之间;检测灵敏度达102拷贝/μl。多重与单重qPCR检测同一样本的相关性较好(R2>0.95)。三重与单重qPCR(t值为1.420~25.786)、四重与单重qPCR(t值为2.505~4.851)检测结果间的差异均无统计学意义(P值均>0.05)。结论建立的多重qPCR可同时对3种以上目的基因进行分型和定量检测,为未来多价轮状病毒疫苗及混合病毒样本中毒株的快速分型和定量检测方法的建立提供了借鉴。
简介:摘要目的探讨1个遗传性血尿肾病耳聋综合征(Alport综合征)家系的致病原因。方法应用高通量测序和Sanger测序法检测先证者COL4A5基因的变异,并在家系其他成员及100名正常对照中进行Sanger测序验证。结果测序结果显示该家系中的4例女性患者COL4A5基因均存在c.3293G>T(p.Gly1098Val)的杂合变异,2例男性患者为COL4A5基因c.3293G>T(p.Gly1098Val)变异半合子,而正常家系成员以及100名正常对照中均未检测到此变异。按照美国医学遗传学与基因组学学会遗传变异分类标准与指南,COL4A5基因c.3293G>T变异位点判读为致病性变异(PM1+PM2+PP1-S+PP3+PP4)。结论COL4A5基因的c.3293G>T(p.Gly1098Val)变异会导致Gly-X-Y三联结构发生变化,可能是该家系的发病原因,新变异的检出丰富了COL4A5基因的变异谱。
简介:摘要目的观察改良25 G硅油取出术的临床效果。方法临床病例对照研究。回顾性分析2017年1月至2018年6月在长春爱尔眼科医院行硅油取出术62例(62眼)的临床资料,所有患者分为常规组32例(32眼)应用25 G硅油取出针头取硅油;改良组30例(30眼)不使用硅油取出针头,直接将连接硅油注吸系统的普通10 ml注射器套入25 G灌注导管取出硅油。记录两组硅油取出术耗时。随访3个月。结果硅油均顺利取出,常规组耗时18.0~39.0分钟,平均(24.50±3.50)分钟。改良组耗时11.0~19.0分钟,平均(12.50±4.50)分钟。两组差异有统计学意义(t=2.753,P=0.021)。随访期间均未发生视网膜脱离等并发症。结论改良25 G硅油取出术安全省时、有效。
简介:摘要目的评价最高序贯脏器衰竭(SOFA)评分、平均SOFA评分用于革兰阳性(G+)/革兰阴性(G-)主要致病菌所致脓毒症患者临床结局预测的价值。方法纳入石河子大学医学院第一附属医院急诊ICU、综合ICU 2016年10月至2017年7月收治的脓毒症患者,于入院时即刻留取血培养、痰培养,分离致病菌,将培养结果阳性患者按照病原学检测结果分为G+和G-致病菌2组。在入院时、24 h、48 h、72 h、4 d、5 d......出院/死亡时间点,采集患者呼吸系统(PO2/FiO2)、肝脏(总胆红素)、肾脏(肌酐)、心血管系统(平均动脉压)、血凝(血小板计数)、神经系统(GCS评分)指标,行SOFA评分,计算平均SOFA评分。结果纳入脓毒症患者62例,其中女20例,男42例,平均年龄(65.79±14.17)岁。G+组28例,G-组34例。患者器官衰竭数目与最高SOFA评分明确相关(G+组男性患者相关系数为0.927,G-组女性患者相关系数为0.847,P<0.05;G-组男性患者关系数为0.719,P<0.01)。最高SOFA评分与患者死亡结局明确相关(G+组相关系数为0.480,G-组相关系数为0.621,P<0.01)。绘制受试者工作特征曲线(ROC),各组最高SOFA评分ROC的曲线下面积(AUC)均高于平均SOFA评分ROC的AUC。结论最高SOFA评分与患者器官衰竭数目、患者死亡结局均相关,最高SOFA评分对于患者死亡结局的预测价值更大。