MLPA在常染色体显性遗传多囊肾病基因诊断中的应用

目的探讨多重连接探针扩增技术(MLPA)在常染色体显性遗传多囊肾病(ADPKD)基因诊断中的应用。方法采用MLPA对20例ADPKD患者的PKD1基因和PKD2基因进行检测。MLPA检测结果显示单一外显子扩增或疑似扩增者采用RT-PCR检测验证;MLPA检测结果显示单一外显子缺失或疑似缺失者采用PCR检测验证,存在扩增产物者进行测序验证。结果 MLPA检测显示:1例患者单一外显子缺失(PKD1 Exon40),5例单一外显子疑似缺失(PKD1 Exon1、PKD1 Exon25、PKD2 Exon8、PKD2 Exon8、PKD1 Exon25),3例单一外显子疑似扩增(PKD1 Exon6、PKD1 Exon7、PKD1 Exon7)。经RT-PCR检测验证,1例患者单一外显子扩增(PKD1 Exon6);经PCR检测与测序验证,1例患者单一外显子错义突变(PKD1 Exon40),1例单一外显子缺失(PKD2 Exon8)。结论 MLPA为ADPKD的基因诊断提供了一种新的方法。     

常染色体显性多囊肾基因PKD1单拷贝区突变

常染色体显性遗传多囊肾病(autosomal dominant polycystic kidney disease,ADPKD)是临床常见的肾脏遗传病之一,目前已发现引起成人ADPKD的基因至少有PKD1和PKD2两种,其中约85%的患者由PKD1基因突变引起。本研究以67例患者PKD1基因单拷贝区的检测为基础,结合PKD1突变检测的所有     

长链PCR在中国汉族人多囊肾病1型致病基因突变检测中的应用

目的:研究特异性分离中国汉族人多囊肾病1型致病基因(polycystic kidney disease gene 1,PKD1)多拷贝区的方法,以排除同源序列对该基因突变检测的干扰.方法:利用与同源序列间的个别碱基差异设计8对能涵盖PKD1多拷贝区的长链PCR引物,分别对两例健康汉族人基因组DNA进行PCR,其扩增产物通过巢式PCR进行序列测定.结果:通过优化PCR体系,尤其是以高质量基因组DNA为模板,适当提高退火温度,经巢式PCR测序证实扩增产物序列与PKD1多拷贝区一致.结论:该研究采用的长链PCR体系可克服同源序列的影响,适用于中国汉族人PKD1多拷贝区的特异性扩增,为进一步通过单链构象多态性分析检测汉族人PKD1突变位点奠定基础.     

上海市汉族常染色体显性多囊肾病遗传异质性和表型差异的研究

目的应用与多囊肾病基因1（PKD1）和多囊肾病基因2（PKD2）紧密连锁的微卫星DNA进行家系连锁分析,研究中国上海市汉族该病的遗传异质性,并比较1型和2型常染色体显性多囊肾病（ADPKD）患者临床表现的差异.方法根据知情同意的原则,收集43个诊断明确的汉族ADPKD家系,利用聚合酶链反应扩增与PKD1和PKD2紧密连锁的微卫星DNA,对PCR扩增产物进行基因扫描分型和连锁分析,确定患病家系与PKD1或PKD2连锁.收集家系中所有患者（包括死亡个体）的病史,现有症状和超声检查结果,对两型患者的临床表现进行统计学比较.结果在进入研究的43个家系中,36个家系与PKD1连锁,占全部家系的84%,7个家系与PKD2连锁,占16%.1型和2型ADPKD患者的诊断平均年龄为36.8岁和46.3 岁,高血压的发生率和诊断平均年龄分别为62%、54%和37.4岁、49.1岁,终末期肾病（ESRD）的发生年龄为51.2岁和64.7岁.结论在上海市汉族中,约84%的ADPKD患者由PKD1突变所致,约16%的患者由PKD2突变所致.两型患者临床并发症发生率无明显差别.但1型患者与2型患者比较,其ADPKD诊断年龄、高血压诊断年龄和终末期肾病发生的平均年龄分别提早9.5年、11.7年和13.5年,这提示1型患者的预后较差.     

129个常染色体显性遗传性多囊肾病家系的致病基因检测及分析

目的 探讨中国常染色体显性遗传性多囊肾病（ADPKD）患者多囊肾病1型致病基因（PKD1）和多囊肾病2型致病基因（PKD2）的突变类型。方法 采用长链PCR和高通量测序方法对129个ADPKD家系的PKD1和PKD2基因进行突变分析,并用双脱氧链终止法测序技术对阳性突变进行验证。结果 在129个ADPKD遗传家系中共检测到116个家系存在PKD1或PKD2基因的118个突变位点,检出率为89.9%（116/129）。PKD1和PKD2的突变率分别为92.2%（107/116）和8.6%（10/116）。在这118个突变位点中,80个（67.8%）为新突变,38个（32.2%）为已知突变;109个位于PKD1（33个已知突变和76个新突变）,9个位于PKD2（5个已知突变和4个新突变）。结论 新发现的PKD1和PKD2突变位点将有助于ADPKD患者的早期诊断和预后预测,并为临床干预提供基本的遗传信息。     

常染色体显性遗传性多囊肾病基因型与临床表型的关系

目的:研究华东地区汉族人常染色体显性遗传性多囊肾病(ADPKD)基因型与临床表现型的关系.方法:利用聚合酶链反应-单链构象多态性分析方法对79例ADPKD患者进行基因诊断,收集经基因诊断确定为2型或1型ADPKD患者的临床资料,以发病年龄、肝囊肿、高血压、尿路感染、尿路结石、血尿等主要临床症状为参数,通过统计学方法研究该病基因型与临床表现型的关系.结果:2型患者的平均发病年龄比1型患者晚9.7岁[(46.09±9.86)岁 vs (36.38±11.35)岁],两者有显著性差异(P<0.05),主要临床症状的出现在两组间无显著性差异.结论:ADPKD2型患者与1型患者临床症状相似,但发病较晚.     

PKD2基因在正常人和2型常染色体显性遗传性多囊肾病患者肾组织中的表达

目的 :观察 PK D2基因在正常人和 2型常染色体显性遗传性多囊肾病 (ADPKD)患者肾组织中的不同表达 ,探讨多囊肾病的发病机制。 方法 :抽提正常人肾组织细胞总 RNA ,通过 RT- PCR法获得 PK D 2基因第 12～ 13外显子 c DNA片段 ,以此为探针 ,用地高辛标记 ,对正常人和 2型 ADPKD患者肾组织分别进行原位杂交 ,并结合图像分析系统观察 PK D2基因表达情况。 结果 :正常人肾组织中 PK D2基因在 Henle襻的厚升支、远曲小管和皮质集合管有较强的表达 (平均光密度为 1.2 3± 0 .0 4 ) ;而 2型ADPKD患者肾组织中 PKD2基因仅在部分囊壁中有少量表达 (平均光密度为 0 .5 6± 0 .0 3)。 结论 :正常人肾组织中 PK D2基因表达量明显高于 2型 ADPKD患者 ,提示 PK D2基因表达降低在 2型 ADPKD的发生和发展中起着一定的作用     

两例结节性硬化症的基因突变分析

目的检测两例中国汉族结节性硬化症散发病例的基因突变位点。方法采用聚合酶链反应扩增结节性硬化症患者、患者家庭中的正常人及100例健康对照的TSC1和TSC2基因的全部外显子,并进行DNA序列分析。结果患者ⅠTSC2基因第268位碱基由胞嘧啶（C）变成胸腺嘧啶（T）,导致第90位氨基酸谷氨酰胺处提前出现终止密码子;患者ⅡTSC2基因第5227位碱基由胞嘧啶（C）变成胸腺嘧啶（T）,导致第1743位精氨酸被色氨酸替代;而患者家庭中的正常人和100例健康对照均无此改变。结论无义突变c.268C〉T和错义突变c.5227C〉T可能是导致这两例患者临床表型的主要原因。     

常染色体显性遗传性多囊肾病研究的热点问题

常染色体显性遗传多囊肾病（ADPKD）是一种常见的遗传性疾病，发病率约为1／1000－1／400。ADPKD致病基因有2个：PKD1和PKD2,分别于1994年和1996年被克隆。目前研究热点主要集中于致病基因表达产物，即多囊蛋白1和多囊蛋白2的结构功能，亚细胞定位，纤毛在多囊肾病发病中的作用，多囊蛋白与血管异常的关系，用影像学方法评价多囊肾病进展，阻断肾素－血管紧张素系统延缓多囊肾病进展以及其他新的诊疗措施等方面。这些问题的深入研究将有助于阐明ADPKD的分子发病机制，为临床彻底治愈ADPKA奠定基础。     

多囊肾病高血压的发病机制与诊治进展

PKD是人类最常见的单基因遗传性肾脏疾病,按其遗传方式可分为常染色体显性多囊肾病( ADPKD)和常染色体隐性多囊肾病(ARPKD),其中ADPKD是发病率最高的遗传性肾病,约占透析人群的5％,是终末期肾脏病的第4位病因.临床上PKD引起的诸多并发症中以高血压最为常见,也是PKD最早期的临床表现.PKD患者的血压增高要比普通人群早十多年,在肾功能衰竭发生前,高血压的发生率约为70％～80％,若进入终末期肾脏病阶段,则几乎所有患者均伴有高血压.     

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.