Mutation analysis and prenatal diagnosis of EXT1 gene mutations in Chinese patients with multiple osteochondromas

Background  Multiple osteochondromas (MO), an inherited autosomal dominant disorder, is characterized by the presence of multiple exostoses on the long bones. MO is caused by mutations in the EXT1 or EXT2 genes which encode glycosyltransferases implicated in heparin sulfate biosynthesis.

Methods  In this study, efforts were made to identify the underlying disease-causing mutations in patients from two MO families in China.

Results  Two novel EXT1 gene mutations were identified and no mutation was found in EXT2 gene. The mutation c.497T>A in exon 1 of the EXT1 gene was cosegregated with the disease phenotype in family 1 and formed a stop codon at amino acid site 166. The fetus of the proband was diagnosed negative. In family 2, the mutation c.1430-1431delCC in exon 6 of the EXT1 gene would cause frameshift and introduce a premature stop codon after the reading frame being open for 42 amino acids. The fetus of this family inherited this mutation from the father.

Conclusions  Mutation analysis of two MO families in this study demonstrates its further application in MO genetic counseling and prenatal diagnosis.

     

裸项栉鰕虎鱼感染创伤弧菌初报

目的 分离及鉴定感染实验用鰕虎鱼的病原菌。方法 将病料分离培养,纯化培养后,用16s rRNA及生理生化方法鉴定病原菌。结果 分离到了革兰阴性弧菌,命名为：GDLAMI-1210株,显微镜下观察为逗点样形态,电镜下可看到菌的一端钝圆,一端长杆弯曲的鞭毛,生理生化鉴定结果与弧菌一致,16s rRNA测序结果与创伤弧菌标准菌株（ATCC27562T）等聚为一类菌,回归实验证实该菌对裸项栉鰕虎鱼有一定的致病性。结论 裸项栉鰕虎鱼感染创伤弧菌初报。     

2型糖尿病患者胰高糖素受体基因Gly40Ser突变的检测

为探讨中国人２型糖尿病与胰高糖素受体（ＧＣＧ－Ｒ）基因４０号密码子的错义突变（Ｇｌｙ４０Ｓｅｒ）是否存在关联,运用聚合酶链反应－限制性片段长度多态性（ＰＣＲ－ＲＦＬ－Ｐ）分析方法在１６０例无亲缘关系之中国云南昆明地区汉族人（２型糖尿病患者１００例,健康对照者６０例）中对ＧＣＧ－Ｒ基因Ｇｌｙ４０Ｓｅｒ突变进行了检测。结果显示：所有研究对象中无论是２型糖尿病患者还是健康对照者均无一例存在ＧＣＧ－Ｒ基因     

Comparison of IS6110 and ‘short fragment’ devR (Rv3133c) gene targets with phenotypic methods for diagnosis of Mycobacterium tuberculosis

Background

Tuberculosis (TB), declared a global emergency in 1993 by the WHO, remains a worldwide public health problem. Rapid diagnosis is required for treatment and prevention. This study compares genotypic methods using two gene targets [IS6110 and 'short fragment' devR (Rv3133c)] with phenotypic methods [Lowenstein Jensen (LJ) and BACTEC 460] for the diagnosis of TB while using Ziehl Neelsen (ZN) staining as the gold standard.

Methods

56 clinical TB samples from a tertiary care apex center along with 50 healthy control samples, excluding samples from patients already on ATT were processed by routine USP methodology. Smears were graded by ZN stain. Solid media (LJ) and liquid media (BACTEC 460) were used along with IS6110 and 'short fragment' devR (Rv3133c) specific gene amplification and comparatively analyzed.

Results

50/56 samples were positive by phenotypic methods, 53 by IS6110 and 45 by devR (Rv3133c) amplification. 38 samples were positive by both phenotypic and genotypic methods. IS6110 detected six and devR (Rv3133c) detected five phenotypically negative samples. Both IS6110 and devR (Rv3133c) were positive in 42 samples. 11 devR (Rv3133c) negative samples were positive by IS6110 and three IS6110 negative samples were positive by 'short fragment' devR (Rv3133c). Compared to phenotypic methods, the sensitivity, specificity, positive and negative predictive values of IS6110 was 94%, 89.29%, 88.68% and 94.34% while that of devR (Rv3133c) was 80%, 91.07%, 88.89% and 83.61% respectively.

Conclusion

Simultaneous use of both phenotypic and genotypic methods increases the yield of positive results.     

Identification of Two Disease-causing Genes TJP2 and GJB2 in a Chinese Family with Unconditional Autosomal Dominant Nonsyndromic Hereditary Hearing Impairment

Background:

There are more than 300 genetic loci that have been found to be related to hereditary hearing impairment (HHI), including 92 causative genes for nonsyndromic hearing loss, among which 34 genes are related to autosomal dominant nonsyndromic HHI (ADNSHHI). Traditional linkage analysis and candidate gene sequencing are not effective at detecting the ADNSHHI, especially for the unconditional families that may have more than one pathogenic cause. This study identified two disease-causing genes TJP2 and GJB2 in a Chinese family with unconditional ADNSHHI.

Methods:

To decipher the genetic code of a Chinese family (family 686) with ADNSHHI, different gene screening techniques have been performed, including linkage analysis, candidate genes screening, high-throughput sequencing and Sanger sequencing. These techniques were done on samples obtained from this family over a period of 10 years.

Results:

We identified a pathogenic missense mutation, c. 2081G>A (p.G694E), in TJP2, a gene that plays a crucial role in apoptosis and age-related hearing loss (ARHL). The mutation was co-segregated in this pedigree in all, but not in the two patients who presented with different phenotypes from the other affected family members. In one of the two patients, we confirmed that the compound heterozygosity for p. Y136* and p.G45E in the GJB2 gene may account for the phenotype shown in this patient.

Conclusions:

We identified the co-occurrence of two genetic causes in family 686. The possible disease-causing missense mutation of TJP2 in family 686 presents an opportunity for further investigation into ARHL. It is necessary to combine various genes screening methods, especially for some unconventional cases.     

金黄地鼠与白化地鼠遗传生化基因位点概貌

目的 建立金黄地鼠和白化地鼠遗传生化基因位点.方法 选用小鼠和大鼠的遗传生化基因位点,采用蛋白质和同工酶醋酸纤维电泳的方法,对金黄地鼠和白化地鼠进行生化基因位点检测.结果 建立了金黄地鼠和白化地鼠25个生化基因位点,分析金黄地鼠和白化地鼠遗传生化基因位点的多态性,为进一步研究金黄地鼠白化突变系的遗传机理奠定基础.结论 金黄地鼠生化基因位点存在多态性,白化地鼠与金黄地鼠生化基因位点存在差异.     

高原人群CYP2C9和VKORC1基因多态性及其与华法林用药剂量的相关性研究

目的 探讨高原人群基于中国人群华法林用药剂量计算公式（PRC模型）和国际华法林遗传药理学协会推荐的亚裔人群华法林剂量计算公式（IWPC模型）两种模型预测剂量的准确性及其临床应用价值。方法 回顾性分析曲靖市第一人民医院2016年10月—2020年1月375例行华法林代谢基因多态性检测患者的临床资料。通过qRT-PCR检测CYP2C9和VKORC1基因多态性,记录患者基本信息和临床用药情况,采用两种模型计算预测剂量并分析其与维持剂量［国际标准化比值（INR）稳定维持在2.0～3.0范围内时所服用的华法林剂量］的相关性,评估两模型预测的准确性。结果 在实际治疗过程中患者是否选择服用华法林进行抗凝,通常与患者的性别、身高、吸烟史、是否合并房颤、是否注射低分子肝素钙无关,而与患者年龄、体重、体表面积（BSA）、初INR、是否置换主动脉瓣膜、是否服用阿司匹林、利伐沙班或氯吡格雷相关。375例患者CYP2C9和VKORC1基因频率符合Hardy-Weinberg遗传平衡定律,其中CYP2C9基因*1/*1（AA）、*1/*3（AC）及*3/*3（CC）基因型频率分别为93.07%（349/375）、6.93%（26/375）和0.00%（0/375）,VKORC1-1639基因AA、AG和GG基因型频率分别为82.66%（310/375）、16.27%（61/375）和1.07%（4/375）。无论是使用PRC还是IWPC模型,除CYP2C9*1/*1 & VKORC1AA（n =289）组与CYP2C9*1/*3 & VKORC1AG（n =5）组预测剂量的差异无统计学意义（P >0.05）外,其他所有基因型组预测剂量两两比较均有差异（P <0.05）。在收集到维持剂量的174例患者中,CYP2C9*1/*1 & VKORC1 AG和CYP2C9*1/*1 & VKORC1 GG组维持剂量分别为［（3.41±1.01）mg,n =30］和［（4.75±0.35）mg,n =2］,均高于CYP2C9*1/*1 & VKORC1 AA组［（2.59±0.73）mg,n =136］（P <0.05）;CYP2C9*1/*3 & VKORC1 AA组维持剂量为［（2.00±0.53）mg,n =5］,低于其他基因型组合（P <0.05）。PRC和IWPC模型预测准确性分别为72.99%（127/174）和62.64%（109/174）;Pearson相关系数（r₁ =0.546,r₂ =0.567）;决定系数（R₁²=0.298,R₂²=0.322）。两个模型预测剂量间无差异（r =0.839,P >0.05）。结论 携带CYP2C9*3等位基因的患者对华法林更敏感,所需华法林剂量较低,而携带VKORC1-1639 G等位基因的患者需要更高剂量的华法林才可以获得有效的抗凝疗效。基因组学预测剂量可为临床使用华法林剂量提供依据,提高抗凝治疗的安全性和有效性。     

Prevalence of Helicobacter pylori, cytomegalovirus,and Chlamydia pneumoniae immunoglobulin seropositivity in coronary artery disease patients and normal individuals in North Indian population

Background

In present day atherosclerosis is perceived as a chronic inflammatory vascular condition and infectious diseases are believed to contribute to its pathophysiology. In this context, the microorganisms which are believed to play a role in the pathophysiology include Chlamydia pneumoniae, cytomegalovirus (CMV), and Helicobacter pylori.

Method

A case control study (retrospective) was conducted over a two-year period. The study population was divided into two groups with 200 individuals in each group. The first group comprised cases of coronary artery disease (CAD) and the second comprised healthy controls selected from the general population after matching for age and sex. Enzyme-linked immunosorbent assay (ELISA) was done for immunoglobulin (IgG) antibodies to H. pylori, C. pneumonia, and CMV. They were also evaluated for conventional risk factors including hypertension, diabetes, obesity, and dyslipidaemia. Epi Info™ version 6 six software was used for analysis of data. Odds ratio, χ² for trend and multiple logistic regression analysis were used to find out statistically significant results.

Results

Seropositivity for H. pylori was present in 119 patients of CAD (59.5%) but it was present in only 76 controls (38%) (P = 0.001). There was a statistically significant association between seropositivity for H. pylori and CAD. There was no statistically significant association between C. pneumoniae and CMV seropositivity with CAD. Multiple logistic regression analysis was done with CAD as the outcome (dependent variable). The predictor covariates (independent) variables were seropositivity to H. pylori, C. pneumoniae, and CMV, hypertension, obesity, diabetes, and dyslipidaemia. It was found that seropositivity to H. pylori, hypertension, obesity, and dyslipidaemia were significant risk factors for CAD.

Conclusion

Our study shows an association between IgG antibody response to H. pylori and CAD. Multiple logistic regression analysis showed that this association was retained even on comparison with the other risk factors.     

HPLC法测定稳心颗粒中三七皂苷R1、人参皂苷Rg1、Rb1、Rd与党参炔苷的含量

[目的] 建立高效液相法（HPLC）同时测定稳心颗粒中三七皂苷R₁、人参皂苷Rg₁、Rb₁、Rd与党参炔苷的含量。[方法] 采用Agilent1260 DAD高效液相色谱仪,Amethyst C₁₈色谱柱（4.6 mm×250 mm,4μm）,乙腈-水为流动相,检测波长为210 nm,流速1 mL/min。[结果] 三七皂苷R₁对照品在8~247 mg/L线性关系良好,平均回收率为102.69%,RSD=2.74%（n=6）;人参皂苷Rg₁对照品在14~422 mg/L线性关系良好,平均回收率为98.72%,RSD=1.85%（n=6）;党参炔苷对照品在1.5~42 mg/L线性关系良好,平均回收率为97.69%,RSD=1.94%（n=6）;人参皂苷Rb₁对照品在2.65~847 mg/L线性关系良好,回收率为102.11%,RSD=1.96%（n=6）;人参皂苷Rd对照品在8~255 mg/L线性关系良好,平均回收率为99.66%,RSD=2.49%（n=6）。[结论] 本实验中三七皂苷R₁、人参皂苷Rg₁、Rb₁、Rd和党参炔苷的含量测定方法稳定可靠,可作为稳心颗粒中三七皂苷R₁、人参皂苷Rg₁、Rb₁、Rd与党参炔苷含量测定方法。     

对氧磷酶1和脂蛋白磷脂酶A2与冠心病的相关性

目的 探讨冠心病患者血清对氧磷酶1(PON1)与脂蛋白磷脂酶A₂(LP-PLA₂)活性与冠脉病变支数及斑块稳定性的相关性.方法 入选88例冠心病患者为研究组,同期行冠状动脉造影术(CAG)排除冠心病的32例患者为对照组.检测两组患者血清PON1、LP-PLA₂活性,对PON1、LP-PLA₂活性与冠状动脉病变支数及斑块稳定性进行相关性分析.结果 冠心病患者血清PON1活性明显低于对照组(P<0.05);活性均值随冠状动脉病变支数增加而降低(P<0.05); 在斑块不稳定组PON1活性明显低于斑块稳定组(P<0.05).冠心病患者血清LP-PLA₂活性均值明显高于对照组(P<0.05),随冠状动脉病变支数增加而升高(P<0.05),且与患者总胆固醇、低密度脂蛋白水平呈正相关.结论 血清PON1及LP-PLA₂活性与冠心病的发生及冠状动脉病变支数有相关性,PON1活性降低可作为冠状动脉斑块趋于不稳定的观察指标.     

Correlation of histomorphologic prognostic markers and proliferative index with loss of heterozygosity 1p/19q and MGMT status in diffusely infiltrating gliomas

Background

Loss of heterozygosity (LOH)1p/19q, and epigenetic silencing of O⁶-methylguanine-DNAmethyltransferase (MGMT) gene, displayed promising role as predictive and prognostic markers in brain tumours. The present study correlated both with conventional histomorphologic prognostic markers and proliferative index in diffusely infiltrating gliomas (DIG).

Methods

Tissues from 45 patients were evaluated for LOH1p/19q using polymerase chain reaction based microsatellite analysis; and for MGMT using immunohistochemistry. Results were correlated with age, histologic type, WHO grade, and proliferation index.

Results

Mean MIB-1 LI showed significant correlation with tumour grade. MGMT-staining in grade II and IV tumours were 31.1% and 16.8%, respectively, while in DA and GBM it was 88.2% and 19.0%, respectively, which were statistically significant. Sixteen cases showed LOH 1p and/or 19q of which 10 (5 oligodendroglial, 3 GBM, AA, DA) had combined LOH; while three each showed 1p (all GBM) and 19q (2 DA and GBM) loss. In the MIB-1LI ≤ 5% and >5% groups LOH 1p and/or 19q was encountered in 6 and 10 cases, respectively. A significant inverse association was noted between LOH with MGMT.

Conclusions

LOH1p/19q and MGMT shows good correlation with conventional histomorphologic and proliferation markers, and should constitute part of the optimal diagnostic workup of DIG.     

香附对川芎中阿魏酸在正常及偏头痛模型大鼠体内药代动力学的影响

目的：研究香附对川芎中阿魏酸（FA）在正常及偏头痛模型大鼠体内的药代动力学影响。方法：用硝酸甘油制作偏头痛大鼠模型,给正常及模型大鼠灌服川芎和川芎-香附合煎液,采用HPLC法检测给药后不同时间大鼠血浆中FA的浓度,DAS2.0软件计算药代动力学参数。结果：正常组中川芎配伍香附后FA的吸收半衰期t1/2Ka明显缩短（P<0.05）,达峰时间tmax提前（P<0.05）,但其他参数与配伍前并无差异。模型组中川芎配伍香附后FA的Cmax、药时曲线下面积AUC(0-t)、分布半衰期t1/2α、表观分布容积V1/F 显著增大（P<0.01）,达峰时间tmax提前,清除率CL/F显著降低（P<0.01）。结论：在正常大鼠体内,香附加快了FA的吸收,而在偏头痛模型大鼠体内,香附能减慢FA在体内的分布和代谢,延长药物成分在体内的作用时间,从而提高了FA的生物利用度。     

双向选择标记amdS在黑曲霉中自发突变及amdS-检测

目的将传统上作为营养缺陷标记的amdS进行诱变,使其成为双向选择标记,有利于构建黑曲霉多基因缺失菌株。方法以已有amdS作选择标记的黑曲霉双基因缺失株为出发突变株,在以尿素为唯一氮源的氟代乙酰氨培养基上筛选amdS-突变菌株。结果将另一以amdS作选择标记的基因进行转化,得到一株三基因缺失菌株。结论在黑曲霉中利用amdS可作为双向选择标记的特点,构建多基因缺失菌株是行之有效的。     

内蒙古地区妇女散发性乳腺癌BRCAl基因突变分析

目的：筛查内蒙古地区散发性乳腺癌患者中BRCAl突变位点。方法：收集新近确诊的乳腺癌患者病例及全血标本40例。以及非癌乳腺良性疾病患者全血30例作为对照,利用TIANampBloodDNA Kit试剂盒获取基因组DNA;PCR方法扩增基因组DNA的BRCAl基因第11外显子的2620～2894bp和3570～3847bp片段．所有标本利用ABl3100-DNA测序仪分析扩增的DNA片段。结果：测序的DNA片段与GeneBank中的序列进行了比对,在所扩增DNA片段中未发现碱基的突变。结论：在我们的研究中所检测的散发性乳腺癌患者的BRCAl基因第11外显子2620～2894bp和3570～3847bp区域未发现突变。     

原位合成四氧化三锰处理模拟核电厂含Co2+放射性废水

在硼酸体系中,以⁵⁹Co作为模拟非放射性同位素研究了原位合成四氧化三锰处理模拟核电厂放射性废水中Co²⁺的工艺条件。考察了反应时间、n(Mn²⁺):n(Co²⁺)、空气流量、反应温度以及pH对出水Co²⁺质量浓度的影响,并由正交试验L₉(4³)优化工艺条件。研究表明:在废水Co²⁺初始质量浓度10mg/L,硼酸质量浓度(以B计)1000mg/L条件下,最佳工艺条件为反应时间105min、n(Mn²⁺):n(Co²⁺)=25:1、空气流量0.7L/min、反应温度65℃以及pH10.5,在此条件下出水Co²⁺质量浓度约为5.68ng/L,去除效率大于99.99%,产物经XRD分析证明沉渣为Mn₃O₄和CoMn₂O₄混合物。