上海地区葡萄糖-6-磷酸脱氢酶基因突变分析

目的 :了解上海地区葡萄糖 6 磷酸脱氢酶 (G6PD)缺乏的基因突变型及其主要临床表现。　方法 :观察 45例上海地区G6PD缺乏患者的主要临床表现 ,测定红细胞G6PD活性并用PCR 限制性内切酶酶解分析法进行G6PD基因突变分析。　结果 :共发现了五种基因突变型 ,分别是cDNA1376G→T(31.2 % )、cDNA1388G→A(2 8.9% )、cD NA95A→G (2 0 .0 % )、cDNA10 2 4C→T(4 .4% )和cDNA493A→G(6 .6 % )。　结论 :上海地区常见的三种G6PD基因型 (cDNA1376G→T ,cDNA1388G→A和cDNA95A→G)占 80 .1% ,酶活性均低于正常人的 10 % ,临床表现以蚕豆病、药物诱发溶血及新生儿期黄疸为主     

海南汉族、黎族人葡萄糖-6-磷酸脱氢酶缺乏症的基因突变

目的:阐明海南汉族、黎族人群中葡萄糖-6-磷酸脱氢酶缺乏症的分子基础。方法:用聚合酶链反应、限制性内切酶消化筛查了1388G→A、1376G→T、1360C→T、1024C→T、592C→T、517T→C、493A→G、487G→A、392G→T和95A→G突变;用单链构象多态性分析筛查其它突变;用核苷酸顺序分析鉴定具有SSCP异常区带样品的突变。结果:在59例汉族G6PD缺乏症患者中,发现1388G→A14例(23.7%)、1376G→T19例(32.2%)、871G→A3例(5.1%)、835A→T1例(1.7%)、517T→C1例(1.7%)、392G→T3例(5.1%)、95A→G4例(6.8%);在32例黎族G6PD缺乏症患者中,发现1388G→A6例(18.8%)、1376G→T18例(56.2%)、871G→A3例(9.4%)、95A→G2例(6.3%);结论:海南汉族、黎族人群中具有共同的常见G6PD基因突变型,提示一些G6PD基因突变是中国南方人群的遗传特征。     

长沙地区婴幼儿G6PD基因突变与急性溶血性贫血关系的研究

目的　了解湖南长沙地区葡萄糖- 6 -磷酸脱氢酶(G6PD)缺乏症的基因突变类型,探讨G6PD缺乏基因突变与婴幼儿急性溶血性贫血的关系。方法　采用突变特异性扩增系统方法对10 3例G6PD缺乏引起的急性溶血性贫血患儿进行G6PD基因突变检测,并对其临床表现进行分析。结果　10 3例G6PD缺乏引起的急性溶血性贫血患儿中检出G1388A2 1例(2 0 .4 % ) ,G1376T4 1例(39.8% ) ,A95G32例(31.1% ) ,未知型9例(8.7% )。各基因型的酶活性比较无显著性差异(P >0 .0 5 ) ,溶血持续天数比较差异无显著性(P >0 .0 5 ) ,各基因型的血红蛋白含量及红细胞数比较差异有显著性(P <0 .0 1) 。结论　G1388A、G1376T、A95G基因突变型是湖南长沙地区最常见的G6PD缺乏基因突变类型,且均可引起婴幼儿急性溶血性贫血,进食蚕豆是重要的诱发因素。三种基因型引起的急性溶血性贫血溶血程度有差异。     

贵州省江口县土家族葡萄糖-6-磷酸脱氢酶基因突变分析

目的：了解贵州省江口县土家族葡萄糖-6-磷酸脱氢酶（G6PD）缺陷症的基因频率、基因突变类型特点及分布特征,从分子水平揭示G6PD基因多态性。方法：采集世居当地土家族男性血液样本227例,用四氮唑蓝（NBT）纸片定性法及G6PD/6PGD比值法做G6PD缺陷症筛查。确诊者用错配引物介导的聚合酶链反应/限制性内切酶酶切分析法（PCR-RE）进行中国人常见9种G6PD基因突变型分析,突变特异性扩增系统法（ARMS）验证其中3种突变。结果：227名土家族男性中检出17例G6PD缺陷者,总检出率7.49%,G6PD缺陷症基因频率0.0749。基因分析检出cDNA 1388（G→A）12例,在G6PD缺陷者中的基因频率为0.706,未知突变5例,基因频率为0.294。结论：G6PD缺陷症在贵州土家族人群有着较高的基因频率,其主要突变类型为G6PD基因cDNA 1388（G→A）。     

G6PD缺陷症基因型分析：一种新的错义突变

目的 对葡萄糖-6-磷酸脱氢酶(G6PD)缺陷症患者进行G6PD基因型分析,分析基因突变类型.方法 选取经G6PD活性测定确诊为G6PD缺陷症的49例患者和100名G6PD活性正常的体检者的全血标本,PCR和DNA测序法对G6PD基因外显子2～13进行序列分析.结果 49例G6PD缺陷症患者中,共发现12种错义突变,其中常见的三种为G6PD G1388A(26.5%)、G1376T(28.6%)和A95G(14.3%),此三类基因型患者G6PD活性仅为正常人群的5%～18%.临床主要表现为新生儿黄疸、进食蚕豆后发生急性溶血性贫血等.其余突变类型包括C1024T(4.1%)、C1225T(2.0%)、C1159T(2.0%)、G487A(4.1%)、G392T(4.1%)、G1160A(6.1%)、G871A/C1311T(4.1%)和C406T/C1311T(2.0%);在外显子7上发现了一种新的错义突变即G691C,该突变造成G6PD第231位丙氨酸(Ala)被脯氨酸(Pro)替代.正常对照标本未发现相同的基因改变.结论 G6PD基因G1388A、G1376T和A95G是G6PD缺陷症患者最常见的三种突变类型.新发现的G691C突变导致Ala231Pro,是引起红细胞G6PD活性降低,患者临床出现溶血症状的主要原因.     

一种新的G6PD基因突变型的鉴定

目的:鉴定1例葡萄糖-6-磷酸脱氢酶缺乏症患者的基因突变。方法:用聚合酶链反应、限制性内切酶筛查葡萄糖-6-磷酸酶基因1388G→A、1376G→T、1360C→T、1024C→T、592C→T、517T→C、493A→G,487G→A、392G→T、95A→G突变,用单链构象多态性筛查葡萄糖-6-磷酸脱氢酶基因的所有外显子,用核苷酸序列测定确定基因突变。结果:该患者未存在1388G→A、1376G→T、1360C→T、1024C→T、592C→T、517T→C、493A→G、487G→A、392G→T、95A→G突变,但在外显子8发现了一种新的G6PD基因突变———835A→G突变,此突变导致第279位的苏氨酸被丙氨酸取代,将其命名为G6PD-海口,其酶活性约是正常的10%,比835A→T突变型的活性低,后者的酶活性约是正常的40%;分析人G6PD的三维结构模型表明,第279位苏氨酸残基的羟基对于维持G6PD亚基的相互作用具有非常重要的作用。结论:835A→G突变是一种新的G6PD基因突变型,G6PD的第279位苏氨酸残基的羟基是维持G6PD亚基相互作用及酶活性的必需基团。     

海南省新生儿葡萄糖-6-磷酸脱氢酶缺乏症的基因突变型分析

[目的]通过对新生儿疾病筛查中确诊的葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症患儿进行基因检测,确定海南省新生儿G6PD缺乏症的分子生物学基础.[方法]通过荧光斑点法在新生儿群体中筛查出可疑或阳性者,经G6PD/6GPD比值法确诊.将G6PD缺乏症患儿静脉血做DNA直接扩增法(PCR)、PCR-等位基因特异的寡核苷酸探针(PCR-ASO)分析、PcR-限制性片段长度多态性(PCR-RFLP)分析和PCR-单链构型多态性(PCR-SSCP)分析,检测基因突变.[结果]G6PD缺乏症新生儿51例中属于G1 376T的16例(31.37%)、属于G1 388A的11例(21.57%)、属于A95G的4例(7.84%)、属于G392T的2例(3.92%)、属于G871A的2例(3.92%)、属于A835T的1例(1.96%),属于T517C的1例(1.96%);余14例没有发现上述突变.[结论]海南省新生儿G6PD缺乏症的基因突变型与我国该遗传病其他高发地区的基因突变谱相似;未发现A835G-G6PD-海口突变型.     

云南籍葡萄糖-6-磷酸脱氢酶缺乏症基因突变研究

目的研究云南籍葡萄糖-6-磷酸脱氢酶（G6PD）基因突变型特点，探讨检测G6PD缺乏症基因突变型的有效方法．方法应用硝基四氮蓝（NBT）纸片法进行G6PD缺乏症定性筛查，等位基因特异抗突变系统（ARMS），聚合酶链反应-单链构象多态性分析（PCR—SSCP），变性梯度凝胶电泳（DGGE）和DNA测序分析46例云南籍G6PD缺乏症患者的G6PD基因突变类型．结果46例样本经PCR—ARMS法发现nt-1388G→A18例（39．13％），nt-1376G→T2例（4．30％）；经PCR—SSCP法发现有22例样本有电泳迁移率异常，其DNA测序结果与PCR—ARMS法的结果吻合，并发现2例nt-1311c→T；经PCR—DGGE法分析G6PDexon12发现有30例样本发现异常泳动条带，DNA测序证实26例（56．52％）为nt-1388G→A，4例（8．7％）nt—1376G→T．而PCR—DGGE法分析G6PDexon2未发现有异常泳动条带的样本．结论（1）nt—1388G→A、nt—1376G→T是云南省主要的基因突变型也是中国人中最常见的两种突变型，揭示中华民族有着共同的起源；（2）在检测突变的PCR—ARMS法、PCR—SSCP法和PCR—DGGE法三种方法中，以PCR—DGGE法结合DNA测序，阳性检出率高，简便、快捷、灵敏、结果准确可靠．     

贵州土家族葡萄糖-6-磷酸脱氢酶基因突变型

目的研究贵州土家族葡萄糖-6-磷酸脱氢酶(G6PD)基因致病突变型,检测G6PD缺乏症发生率及基因频率。方法通过硝基四氮唑蓝纸片法对2 789例土家族男性进行G6PD缺乏症定性筛查,用变性高效液相色谱技术和DNA测序对78例个体进行基因突变型鉴定。结果在2 789例纯系土家族男性中,发现2例酶活性异常。在印江县采集的78例(2例酶活性异常和76例正常)纯系土家族男性血样中,共检出G6PD c.1388G>A突变2例、c.1376G>T突变1例和c.1311C>T/IVS 11+93T>C突变2例,基因突变型分别占2.6%、1.3%和2.6%。结论在贵州土家族人群中发现c.1388G>A、c.1376G>T和c.1311C>T/IVS11+93T>C 3种突变型,是共同存在于中国人群的G6PD基因突变型,中华民族可能源于共同的祖先;贵州省印江县土家族男性G6PD缺乏症的基因频率为6.5%,可为上述地区G6PD缺乏症的防治提供依据。     

G6PD缺陷症基因型分析：一种新的错义突变

目的对葡萄糖-6磷-酸脱氢酶（G6PD）缺陷症患者进行G6PD基因型分析,分析基因突变类型。方法选取经G6PD活性测定确诊为G6PD缺陷症的49例患者和100名G6PD活性正常的体检者的全血标本,PCR和DNA测序法对G6PD基因外显子2~13进行序列分析。结果 49例G6PD缺陷症患者中,共发现12种错义突变,其中常见的三种为G6PD G1388A（26.5%）、G1376T（28.6%）和A95G（14.3%）,此三类基因型患者G6PD活性仅为正常人群的5%~18%。临床主要表现为新生儿黄疸、进食蚕豆后发生急性溶血性贫血等。其余突变类型包括C1024T（4.1%）、C1225T（2.0%）、C1159T（2.0%）、G487A（4.1%）、G392T（4.1%）、G1160A（6.1%）、G871A/C1311T（4.1%）和C406T/C1311T（2.0%）;在外显子7上发现了一种新的错义突变即G691C,该突变造成G6PD第231位丙氨酸（A la）被脯氨酸（Pro）替代。正常对照标本未发现相同的基因改变。结论 G6PD基因G1388A、G1376T和A95G是G6PD缺陷症患者最常见的三种突变类型。新发现的G691C突变导致A la231Pro,是引起红细胞G6PD活性降低,患者临床出现溶血症状的主要原因。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.