正常人DMPK基因三核苷酸CTG重复数目的检测

目的 检测正常人ＤＭＰＫ基因３′非编码区ＣＴＧ三核苷酸ＣＴＧ的分布。方法 采用＾３２Ｐ标记的ＰＣＲ和聚丙烯酰胺凝胶电泳技术,对上海地区５８例正常人该重复序列进行了分析。结果 在分析的５８例正常人１１６条染色体中,共发现１６种等位基因,其中ＣＴＧ拷贝数以１２拷贝的频率最高（３２．８％）,其余依次为１１拷贝（２１．６％）,５拷贝（２０．７％）,１３拷贝（１６．４％）,１４拷贝（３．４％）,１５拷贝（３．４％）和１６拷贝（１．７％）。纯合子１４例,其中５例１２／１２,５例５／５,３例１１／１１,１例１３／１３。杂合率为７５．８％。结论强直性肌营养不良是ＤＭＰＫ基因３′非编码区ＣＴＧ三核苷酸的重复扩增所致。正常人ＣＴＧ的拷贝数为５－４０不等,有种族特殊性和高度多态性。     

强直性肌营养不良基因CTG 复数下SEP,MEP对比分析

目的 探讨强直性肌营养不良（ＤＭ）患者及其家系成员基因ＣＴＧ重复数的变化与体感诱发电位（ＳＥＰ）、经颅刺激运动诱发电位（ＭＥＰ）的比较。方法 用聚合酶链反应（ＰＣＲ）扩增及ＤＮＡ杂交法对５例临床诊断ＤＭ患者及其中３个家系１６名成员进行ＤＭ基因ＣＴＧ重复数和ＳＥＰ、ＭＥＰ测定。结果 １０名正常人ＣＴＧ重复 １９～３０个,ＳＥＰ、ＭＥＰ正常和例。ＤＭ患者ＣＴＧ重复数均在８０个以上,其中２例在１６０５个     

上海地区91名正常人强直性肌营养不良基因CTG重复数测定

目的：研究上海地区正常人强直性肌营养不良（ＤＭ）基因中三核苷酸ＣＴＧ重复数的值。方法：用ＰＣＲ扩增及Ｓｏｕｔｈｅｒｎ杂交法检测９１名正常人ＤＭ基因的ＣＴＧ数。结果：测出７个等位基因，其ＣＴＧ重复数分别为５，１１，１３，１５，１９，２３及３０个，其中以１３的频率最高（８０．２１％）。结论：上海地区９１名正常人的结果与日本人群相似，与欧洲人群及中国成都人群不同。     

Charcot-Marie-Tooth病1A型基因重复诊断研究

目的：探察国人Ｃｈａｒｃｏｔ－Ｍａｒｉｅ－Ｔｏｏｔｈ（ＣＭＴ１Ａ）型患者的１７ｐ１１．２－ｐ１２区基因重复及与临床表现型的关系。方法：对１５个ＣＭＴ１家系的２９名成员进行１７Ｐ１１．２－Ｐ１２区的３个短串联重复序列标记物进行多骤酶链反应－３种标记物的杂合率及多态性分析。结果：１６名ＣＭＴ患者检出ＣＭＴ１Ａ重复者９名（５５％）,１３名无症状家属中检出４名基因重复者,ＲＭ－ＧＴ、Ｄ１７Ｓ１３５７、Ｄ１     

心肌细胞缺氧通过激活AMPK促进GLUT4移位和葡萄糖摄取

目的：探讨心肌缺氧时ＡＭＰ激活的蛋白激酶（ＡＭＰＫ）激活对葡萄糖转运子４（ＧＬＵＴ４）移位和葡萄糖摄取的作用。方法：大鼠心室肌经５００μｍｏｌ／Ｌ腺嘌呤－９－β－Ｄ－阿糖呋喃腺苷（ａｒａＡ）处理后,分别与胰岛素、氰化钾、５－氨基咪唑－４－氨基甲酰－１－β－Ｄ核糖呋喃腺苷（ＡＩＣＡＲ）孵育,用放射性核素分析技术测定其葡萄糖摄取量和ＡＭＰＫ活力,应用Ｗｅｓｔｅｒｎ印迹法分析心肌细胞ＧＬＵＴ４含量。结果：ＡＭＰＫ特异性激活剂ＡＩＣＡＲ和氰化钾可使心肌葡萄糖摄取增加（１倍和１．５倍）,但均受ａｒａＡ抑制。ＡＩＣＡＲ增加心肌ＡＭＰＫ活力和葡萄糖摄取,而ａｒａＡ则有抑制作用。心肌细胞质膜ＧＬＵＴ４分布明显增加而细胞器膜ＧＬＵＴ４分布相应减少。结论：氰化钾所致的心肌缺氧与ＡＩＣＡＲ一样可通过ＡＭＰＫ激活途径,促进ＧＬＵＴ４移     

异基因外周血干细胞的动员,检测及临床移植的应用研究

目的：探讨异基因外周血干细胞移植（ａｌｌｏ－ＰＢＳＣＴ）供者经Ｇ－ＣＳＦ或Ｇ－ＣＳＦ＋ＧＭ－ＣＳＦ动员后,采集的外周血干细胞（ＰＢＳＣ）移植物中的幼稚粒细胞与单个核细胞（ＭＮＣ）、ＣＤ３４＾＋细胞及ＣＦＵ－ＧＭ之间的相关性和移植的剂量标准及临床应用效果。方法：对１１例ａｌｌｏ－ＰＢＳＣＴ供者用Ｇ－ＣＳＦ（９例）或Ｇ－ＣＳＦ＋ＧＭ－ＣＳＦ（２例）进行动员,于动员前及动员后,分别对外周血及ＭＮＣ惧物中的幼稚粒细胞、ＣＤ３４＾＋细胞及ＣＦＵ－ＧＭ进行检测计数,预处理方法主要用大剂量环磷酰胺（ＣＴＸ）＋全身照射（ＴＢＩ）。结果：动员后外周血中的幼稚粒细胞与ＣＤ３４＾＋细胞及ＣＦＵ－ＧＭ同步增加,外周血ＭＮＣ中的幼稚粒细胞数与ＣＤ３４＾＋细胞数及ＣＦＵ－ＧＭ有较好的相关性。１１例患者全部植活和恢复造血功能,并为染色体核型     

脑出血早期血清及脑脊液GOT,LDH,CPK测定的临床意义

目的：探讨脑出血早期血清及脑脊液（ＣＳＦ）中谷草转氨酶（ＧＯＴ）、乳酸脱氢酶（ＬＤＨ）、肌酸磷酸激酶（ＣＰＫ）的含量变化及其临床意义。方法：Ｉ列脑出血患者发病７２小时以内的血清含量,并和３０例健康人对照,８例行侧脑室引流得由引流管取ＣＳＦ同时检测,３０例心电图有缺血表现者中的１０例做血清ＣＰＫ ＭＢ及ＣＰＫ－ＢＢ测定。结果：脑出血组血清含量明显高于对照组,二者有非常显著的差异（Ｐ〈０。．０１）,８     

肉苁蓉总苷对大鼠心肌缺血的保护作用

采用结扎冠状动脉造成大鼠心肌缺血模型,研究了肉苁蓉总苷（ＧＣ）对缺血心肌的保护作用。结扎冠状动脉后,心肌中ＳＯＤ,ＣＰＫ活性降低,ＭＤＡ含量升高,心电图表现为Ｓ－Ｔ段升高,大鼠静脉给予ＧＣ,５ｍｉｎ后再结扎冠脉,结果ＧＣ能明显改善缺血心电图,减小心肌梗死面积,提高心肌组织中的ＣＰＫ活性,但对ＳＯＤ和ＭＤＡ无显著影响,提示ＧＣ具有保护缺血心肌作用。     

聚合酶链反应技术在腓骨肌萎缩症基因诊断中的应用

目的 探讨应用聚合酶链反应（ＰＧＲ）技术建立中国人腓骨肌萎缩症（ＣＭＴ）基因诊断的方法。方法 分别应用ＰＣＲ－双酶切、聚合酶链反应－单链构象多态性分析（ＰＣＲ－ＳＳＣＰ）、聚合酶链反应－变性梯度凝胶电泳（ＰＣＲ－ＤＧＧＥ）或ＰＣＲ直接测序等方法对３２个确诊的ＣＭＴ家系进行了ＰＭＰ２２、ＭＰＺ、Ｇx３２等致病基因的突变检测。结果 ２１．９％的ＣＭＴ家系患者有Ｇx３２、ＭＰＺ和ＰＭＰ２基因的突变。ＰＣＲ－ＳＳＣＰ检测到１０个家系有异常泳带,经ＰＣＲ测序证实有５个为多态;另５个为与疾病相关的致病的点突变,即４个Ｇx３２和１个ＭＰＺ基因的点突变。PCR-双酶切诊断了２个包含ＰＭＰ２２基因在内的大片段重复突变的ＣＭＴ１Ａ型家系。ＰＣＲ－ＤＧＧＥ仅１个家系患者异常泳带,该结果也可经ＰＣＲ－ＳＳＣＲ方法检出。结论 ＰＣＲ－ＳＳＣＰ和ＰＣＲ－双酶切可作为Ｇx３２、ＭＰＺ、ＰＭＰ２２基因的点突变和ＣＭＴ１Ａ的大片段重复突变的初筛的方法,而ＰＣＲ－ＤＧＧＥ方法用于Ｇx３２基因的突变检测不理想,点突变应经ＤＮＡ测序证实。     

急性淋巴细胞白血病微量残留病的PCR监测

①目的探讨急性淋巴细胞白血病（ＡＬＬ）微量残留病（ＭＲＤ）检测的临床意义。②方法应用筑巢式多聚酶链反应（ｎｅｓｔｅｄＰＣＲ）对３２例ＡＬＬ进行Ｔ细胞受体（ＴＣＲ）Ｖδ２Ｄδ３基因重排检测。③结果１８例Ｂ系ＡＬＬ中有１５例（７２．２％）、４例Ｔ系ＡＬＬ中有１例（２５．０％）存在ＴＣＲＶδ２Ｄδ３基因重排。同时对１６例进行３９例次微量残留病动态监测，其中６例ＭＲＤ－ＰＣＲ阴性病人随访５．１７～１６．１７年，无１例复发；１０例ＭＲＤ－ＰＣＲ阳性者，２例分别于阳性后０．２５，１．００年骨髓复发，１例有复发倾向，余７例随访４．３３～６．２５年无复发。④结论ＭＲＤ－ＰＣＲ阴性者预后良好，可望长期生存，治疗时应以ＭＲＤ－ＰＣＲ转阴为停止化疗的可靠指标，对ＭＲＤ－ＰＣＲ阳性者应定期监测ＭＲＤ变化，结合病人情况进行综合评价。     

DNA analysis in a suspected individual with myotonic dystrophy family history and her abortus

Objective To observe trinucleotide repeat number, (CTG)n in the 3’-untranslated region of the myotonic protein kinase (MTPK) gene in a clinically suspected woman with myotonic dystrophy (DM) family history and her abortus, in order to confirm the necessity of exerting antenatal examination in patients or suspected individuals with DM family history. Methods Long Expand TM Template polymerase chain reaction (PCR) system was used to analyze CTG trinucleotide repeat numbers located in the 3’ untranslated region of MTPK on chromosome 19q13.2-3 in both peripheral white cells and muscles of the suspected mother and the other two DM patients in the family. The tissues of her abortus and blood of a health woman were detected, too. Results CTG repeats in both peripheral white cells and muscles of the suspected mother and the tissue of abortus were higher than normal range of CTG repeat number. There is no significant difference between blood and muscle samples. High CTG repeats were detected in blood and muscles of the typical DM members in the family, but in the blood sample of control, CTG repeats is normal. Conclusion CTG trinucleotide analyses and antenatal examination should be done in pregnant with a DM family history, in order to reduce the birth rate of DM offspring.     

广西地区壮族1型糖尿病与CTLA4基因多态性关系的研究

目的：分析CTLA4基因多态性与广西地区壮族1型糖尿病（type l diabetes mellitus,T1DM）的关系.方法：提取37例广西地区壮族T1DM患者和40例正常对照组的外周血白细胞基因组DNA,应用聚合酶链反应检测CTLA4第4外显子的3’非翻译区包含（AT） n[CTLA4 （AT）n]重复序列的特异性等位基因,并对两组CTLA4 （AT）n等位基因频率分布进行比较.结果：广西地区壮族人CTLA4（AT）n基因中,发现有12种等位基因（86～122 bp）.壮族T1DM组88 bp等位基因频率共检出24例（64.9％）,与正常对照组8例（20％）比较明显增高,差异有统计学意义（P＜0.01）.结论：广西壮族T1DM患者与CT-LA4基因多态性明显相关,CTLA4（AT）n 88 bp可能是广西壮族T1DM的易感等位基因.     

Myotonic dystrophies

Myotonic dystrophies or dystrophia myotonica (DM) is a clinical syndrome that includes myotonic dystrophy type 1 (DM1), myotonic dystrophy type 2 (DM2), myotonic dystrophy type 3 (DM3), and so forth. The terminology was recommended by the new nomenclature for myotonic dystrophies of an International Panel for Consensus. Previous studies have shown that DM1 is caused by the expansion of a cytosine-thymine-guanine (CTG) repeat in the DM protein kinase gene on chromosome 19, and DM2 is caused by an expansion of a cytosine-cytosine-thymine-guanine (CCTG) repeat in the zinc finger protein 9 (ZNF9) gene on chromosome 3. Because DM1 and DM2 have very similar clinical presentations, the diagnosis of these two disorders needs to be confirmed by molecular genetic analysis. Recently, DM3 was reported to include a multisystem myotonic disorder with frontotemporal dementia, and a linkage to chromosome 15q21-24. Although the age at onset, disease severity, and cerebral abnormality on a brain magnetic resonance spectrometry may correlate with the number of triplet repeats in the blood cells of DM1, it is too early to reach a conclusion. In Taiwan, the prevalence of DM1 is much lower than in Western countries. Previous studies have shown that the central nervous system symptomatology is correlated mainly with the white matter lesions in the brain MRI, but the CNS manifestations seem unrelated to the numbers of CTG triplet repeats in the blood cells. The inverse correlation between age at onset and CTG repeat length is significant only in patients with small expansions of about 100-250 triplet repeats. Transmission contraction of the repeat size is likely to occur in alleles with large repeats and is associated with paternal transmission. In congenital DM1, individual variability of muscle differentiation does occur, in spite of the same number of CTG repeats in the leukocytes.     

强直性肌营养不良一家系的临床和致病基因初步分析

目的:探讨浙江台州地区一个强直性肌营养不良(myo tonic dystrophy,DM)汉族家系的临床表现和分子遗传学基础.方法:分析 该家系44例成员中8例(包括先证者Ⅲ13)临床确诊为DM患者的临床表现,以及5例患 者和6例无症状成员的肌电图表现,并对7例DM患者(除Ⅱ6外)的DNA样品进行DM 1和DM22个位点PCR扩增、琼脂糖电泳检测,对克隆产物进行测序. 结果: 该家系患者除有肌强直、肌萎缩等表现外;心电图检查:心脏传导阻滞(7/8);裂 隙灯检查:白内障(6/7);肌电图检查:患者组有强直电位发放(5/5),无临床症状成员也 存在肌源性损害(5/6);但该家系无DM1位点(CTG)n和DM2位点(CCTG )n的重复数增加.结论:强直性肌营养不良可能存在新的致病基因位 点.     

冠心病合并糖尿病患者dTP分析及随访

目的 分析冠心病(CHD)合并糖尿病(DM)患者11脱氢血栓素B2/6-酮前列环素比值(11-dh-TXB2/6-k-PGF1a,dTP)及与主要不良心血管事件(MACE)和再住院之间的关系.方法 选择2013年7月至2014年6月270例CHD患者作为研究对象,其中136例非DM患者(非DM组),134例合并DM患者(DM组).记录患者临床情况,测量身高、体质量、血压、心率等指标,完善心电图、超声心动图、冠状动脉造影等检查.测定两组患者11-dh-TXB2和6-k-PGF1a水平,计算dTP值.随访1年,记录患者MACE事件及再次住院情况.Epdate软件建库,SPSS17.0软件进行统计分析.结果 非DM组与DM组dTP分别为1.8±0.6和2.0士0.7,差异有统计学意义(P＜0.05).对于非DM组,hs-CRP、收缩压、舒张压、冠状动脉病变数及严重病变数与dTP相关(P＜0.05).而对于DM组,hs-CRP、血糖、胆固醇水平、冠状动脉病变数及严重病变数与dTP相关(P＜0.05).随访1年,非DM组和DM组患者发生MACE事件分别为33例(24.3％)和44例(32.8％),两组比较差异无统计学意义(P＞0.05).非DM组和DM组再次住院患者分别为12例(8.8％)和24例(17.9％),两组比较差异有统计学意义(P＜0.05).发生MACE的患者与无MACE的患者住院时dTP分别为2.3±0.8和1.8±0.6,差异有统计学意义(P＜0.05).再住院患者与未再住院组患者住院时dTP分别为2.4±1.0和1.9±0.6,差异有统计学意义(P＜0.05).结论 CHD合并DM患者dTP明显增高,提示血小板明显活化,且较高的dTP增加了患者MACE事件及再次住院风险,应该强化抗血小板治疗.     

强直性肌营养不良症的临床、家系和遗传特征分析

目的:分析强直性肌营养不良症(myotonic dystrophy or dystrophia myotonia,DM)的临床、家系和遗传特征,提高对强直性肌营养不良症的认识,为DM的基因诊断和产前诊断提供分子依据.方法:收集2个DM家系,对其临床进行详细分析,用片段分析法对2个DM家系内的4例患者和1个家系成员DM1...     

2型糖尿病合并代谢综合征患者心脏变时性功能变化及意义

目的分析2型糖尿病合并代谢综合征(MS)患者心脏变时性功能改变特点,探讨其与心肌缺血的关系,研究2型糖尿病患者心脏变时性功能相关因素。方法选择200例2型糖尿病患者和50例正常健康者作为研究对象,分别分为MS组、T2DM组和对照组,根据心电图ST-T改变将MS组和T2DM组的研究对象分为心肌缺血组和非心肌缺血组,根据心脏变时性功能将MS组和T2DM组的研究对象分为CI组和变时性功能正常组,进行症状限制性平板心电运动试验并收集临床资料。结果 MS组和T2DM组HRmax、rHR、心率储备、变时性指数均低于对照组,其中MS组最小(P<0.05),心肌缺血组CI率和MS患病率均显著高于非心肌缺血组(P<0.05),CI和MS与心肌缺血相关性有统计学意义(OR=3.225,3.568,P<0.05),血脂异常、高血压、MS是2型糖尿病患者心脏CI的独立危险因素(OR=1.952,1.993,2.685,P<0.05)。结论 2型糖尿病患者心脏变时性功能不全发生心肌缺血风险高,变时性功能可作为反映心肌缺血的指标,MS则是2型糖尿病患者心脏变时性功能不全和心肌缺血的高危因素。     

The early risk stratification of the patients with acute chest pain

Objective: This investigation was designed to stratify patients with acute chest pain based on their symptoms, electrocardiogram (ECG), cardiac injury markers and the number of accompanying traditional risk factors(smoking, obesity, hyperlipemia, hypertension, diabetes), and to assess the effect of the above factors to obtain a risk stratification for patients with chest pain. Methods: We identified 139 patients with acute chest pain, including 45 myocardiac infarction patients, 65 unstable angina patients and 29 chest pain patients without identified acute coronary syndrome(ACS) admitted to our Coronary Heart Center during December 2004 to February 2005. All patients accepted coronary angiography. All data was collected using questionnaires. Based on reported symptom, electrocardiogram (ECG), cardiac injury markers and the number of the accompanying traditional risk factors, we stratified all patients into four groups: Group 1, patients with acute chest pain, ECG changes and abnormal cardiac injury biomarkers. Group 2, patients with acute chest pain and ECG changes(without abnormal cardiac injury biomarkers). Group 3, patients with acute chest pain, normal ECG, normal cardiac injury biomarkers and >2 traditional risk factors. Group 4, patients with acute chest pain, normal ECG and normal cardiac injury biomarkers, but only≤2 traditional risk factors. From this data we examined the difference of ACS incidence in the four groups. Results:After stratification the ACS incidence of the grouped patients in turn was 100%, 84%, 69.6% and 53.3%. The combination of early phase ECG and cardiac injury markers identified 70.9% patients with ACS(the specificity being 90.7%). The mortality of group 3 was higher compared with group 4(69.6% vs 53.3%), however the P value was more than 0.05 and didn’t show significant statistical difference. The correlation analysis found the number of the traditional risk factors had a significant positive correlation (r = 0.202, P = 0.044) with the number of stenosis being more than 50% of the artery diameter. Multiple linear regression showed the hypertension had a significant correlation with the number of the diseased regions(P = 0.014). Conclusions:The risk stratification based on the symptom, ECG, cardiac injury markers and accompanying traditional risk factors is both important and available in practice. It is unsuitable for patients with a normal ECG and cardiac injury markers to differentiate ACS from non-cardiac chest pain relying only on the number of the accompanying traditional risk factors. However we found the number of the risk factors can indicate the disease severity.