江门成人散发急性腹泻者星状病毒的检测及毒株型别分析

目的 研究江门地区成人腹泻者星状病毒感染情况及型别特点.方法 收集江门地区病毒性腹泻流行期散发成人腹泻患者粪便标本,经胶体金法、RT-PCR法分别检测轮状病毒和诺如病毒后,运用实时荧光PCR进行星状病毒检测,并对阳性标本进行RT-nested PCR测序,鉴定型别.结果 56份成人病毒性腹泻粪便标本中,荧光PCR法测出3份星状病毒株(5.4%),经RT-PCR测序1份,序列分析鉴定为HAtsV-4型.结论 江门成人散发病毒性腹泻者存在星状病毒感染,型别为HAstV-4,未发现多致泻病毒混合感染.     

广州儿童秋冬腹泻中人类杯状病毒感染的分子流行病学研究

目的了解广州某医院秋冬季儿童腹泻中人类杯状病毒（HuCV）感染的分子流行病学特点。方法连续2个秋冬季腹泻流行季节收集临床诊断为病毒性腹泻患儿的粪便标本。采用ELISA检测轮状病毒，RT-PCR检测HuCV。部分阳性标本PCR产物进行纯化、测序，结合参考株相应核苷酸序列进行进化分析。结果诺瓦克样病毒（Norwalk-like virus，NLVs）阳性率为12-35％（80／648），扎幌样病毒（Sapporo-like virus，SLVs）阳性率为0．31％（2／648）。NLVs各月阳性率无显著性差异，主要感染2岁以下患儿。2003年流行株为GⅡ-3群和GⅡ-4群，2004年为GⅡ-4群。2株SLVs均为GⅠ-1群。结论NLVs是广州地区秋冬季节儿童腹泻的重要病原体，其流行株为GⅡ-3群、GⅡ-4群病毒株， 但在不同的时间段可能存在不同的流行优势株；广州地区存在SLVs所致感染，与我国报告的SLV基因型不同。     

广州市门诊5岁以下儿童诺如病毒感染基因型分析

目的了解2007-2008年广州市监测点医院门诊5岁以下儿童病毒性腹泻散发病例中诺如病毒感染状况及基因型别。方法收集广州市2007-2008年监测点医院门诊临床诊断为病毒性腹泻的患儿临床资料及粪便标本954份,采用ELISA方法检测诺如病毒、轮状病毒,将诺如病毒抗原阳性的标本用RT-PCR方法进行诺如病毒核酸扩增,将阳性产物回收、纯化并测序,结合GenBank中的相关序列进行型别流行特点分析。结果 954份标本中,ELISA方法检测各种病毒的抗原阳性率为诺如病毒18.8%(179/954),其中07年12.2%(32/262),08年21.2%(147/692),轮状病毒30.8%。诺如病毒腹泻病例发病高峰在8～11月份,2岁以下病例数占全部病例的91.6%,病例集中在6～23月龄。RT-PCR方法检测诺如病毒核酸阳性率为80.4%(144/179),144份诺如病毒核酸阳性标本经测序及与GenBank中的相关序列比对证实有1份是GⅡ2型,其余全部为GⅡ4型。结论诺如病毒是广州市门诊5岁以下儿童病毒性腹泻的常见病原之一,仅次于轮状病毒,常见于2岁以下婴幼儿,2007-2008年流行优势株基因型为GⅡ4型。     

浙江省萧山区轮状病毒A组G型和P型基因同源性比较与进化分析

目的分析浙江省萧山区婴幼儿急性腹泻患者中分离的轮状病毒A组G型、P型的基因序列,探讨该病毒的遗传变异和分子特性。方法收集2010年6～12月浙江省萧山区急性婴幼儿（≤5岁）腹泻患者粪便标本,应用多重PCR技术从临床标本中扩增轮状病毒A组G型、P型基因片段并测序分析,比较基因的亲缘关系并绘制进化树。结果162例粪便样本,ELISA结果为54例阳性。PCR检测结果G1型为32株,G2为5株,G3为9株,P型中,P4为3株,P8为5株,成功地对16例A组轮状病毒基因进行测序。2010年A组轮状病毒基因的核苷酸序列与参考株具有较高的同源性,且A组基因核苷酸序列发生了一定的变异。结论轮状病毒A是引起浙江萧山地区急性腹泻的主要的病原之一,其病原具有基因型别的多样性,Gl型可能是2010年浙江萧山地区流行的优势株。     

急性腹泻患儿诺如病毒检测和分型

目的了解苏州地区急性腹泻患儿诺如病毒的分子特征和基因型别。方法收集苏州地区2009年9月-2010年3月共153份5岁以下急性腹泻患儿粪便或肛拭标本,用多重RT—PCR方法检测诺如病毒,选择部分阳性扩增片段进行基因序列测定,并对序列进行系统进化分析。结果急性腹泻患儿诺如病毒检出率为16．34％（25／153）,均为诺如病毒Ⅱ型,且诺如病毒基因型别均为GⅡ-4／2006b变异株。结论诺如病毒是2009～2010年引起苏州地区急性腹泻患儿重要的病原之一,诺如病毒GⅡ-4／2006b变异株或类似株可能是2009～2010年苏州地区流行的优势株。     

PCR方法在肠传病毒检测中的应用

多聚酶链反应(PCR)扩增DNA技术是科学发展的重大突破。近年来在国内外都有越来越广泛的应用。本研究在国家自然科学基金资助下,对我国一些重要肠传病毒,如轮状病毒、脊灰病毒、甲肝病毒、诺瓦克病毒,分别建立了套式PCR方法区分轮状病毒的G型和P型;RT-PCR方法鉴别脊灰病毒为疫苗株或野毒株;抗体捕捉PCR方法检测临床和环境样品中的甲肝病毒;RT-PCR方法结合扩增产物测序分析首次在我国发现有诺瓦克样病毒感染等。     

东莞市病毒性腹泻病原学分析

目的了解东莞市病毒性腹泻的病原学分布情况。方法收集2010年7月至2012年9月东莞市东华医院感染性腹泻患者的粪便样品,采用ELISA方法检测轮状病毒,采用Realtime RT-PCR方法检测诺如病毒,采用RT-PCR方法检测星状病毒,采用Realtime PCR方法检测腺病毒。结果检测粪便样品共224份,诺如病毒、轮状病毒、腺病毒、星状病毒检测阳性率分别为17.9%、11.6%、4.5%,0.4%,总检测阳性率为31.3%。对其中22份轮状病毒阳性样品进行血清分型,结果G血清型中,G1型10株,G2型1株,G3型8株,G9型3株。P血清型中,P6型1株,P8型12株,9株未能分型。40株诺如病毒全部为GⅡ型。1株星状病毒经测序分析为1型。结论东莞市腹泻病毒主要感染3岁以下的婴幼儿,诺如病毒和轮状病毒为主要病原体,轮状病毒的分布呈多样性,以G1型为主。     

两起诺如病毒感染疫情的分子流行病学特征分析

目的了解广州市海珠区2014年5月和2015年3月发生的两起诺如病毒感染性腹泻疫情的分子流行病学特征。方法采用实时荧光RT-PCR检测肛拭子标本中诺如病毒RNA,选取部分阳性标本进行核酸序列测定并进行同源性分析。结果在这两起疫情采集的61份肛拭子标本中,诺如病毒GⅡ型阳性38份。经测序比对后,2014年5月的流感疫情中检测出1株GⅡ-6型和1株GⅡ-9型;2015年3月的流感疫情中检测出2株GⅡ-6型和1株GⅡ-5型。结论广州市海珠区两起疫情存在两种基因型诺如病毒的混合感染。     

2014年重庆地区511例急性腹泻儿童中诺如病毒腹泻流行特点与分子亚型鉴定

目的 了解2014年重庆地区诺如病毒(norovirus,NV)感染所致儿童急性腹泻的感染现状、流行基因型以及发生基因重组情况.方法 2014年在重庆医科大学附属儿童医院门诊检验科收集符合纳入条件的腹泻标本511例,采用RT-PCR分别对NV GⅠ和GⅡ基因组的衣壳蛋白区进行扩增、测序,并对序列进行基因分型及构建进化树;为进行重组分析,再次对NV阳性标本的RdRp区和capsid区同时进行扩增,并用SimPlot进行重组鉴定.结果 共收集到符合条件的标本511例(男性290例,女性221例),检出NV GⅡ型67例,NV检出率为13.1％;8～10月为NV检出高峰,阳性率最高的年龄组为7 ～12月龄.67例NV阳性标本在capsid区测序成功,检测到5种基因型,依次是GⅡ.4 Sydney 2012型42例(62.7％),GⅡ.3型8例(11.9％)和GⅡ.17型8例(11.9％),GⅡ.6型7例(10.4％),GⅡ.14型2例(3.0％).59例NV病毒阳性标本同时在RdRp区和capsid区测序成功,共检测到52例重组株,4种重组型分别是GⅡ.e/GⅡ.4 Sydney 2012型42例、GⅡ.12/GⅡ.3型6例、GⅡ.7/GⅡ.6型2例、GⅡ.7/GⅡ.14型2例.结论 2014年重庆地区NV检出率为13.1％,GⅡ.4 Sydney 2012为最主要流行基因型,且NV重组现象十分明显.     

2009~2011年大庆地区感染人轮状病毒G血清型鉴定

目的了解大庆地区2009—2011年感染人的轮状病毒流行株的血清型情况。方法采用A组轮状病毒ELISA检测试剂盒对采集来的288例腹泻标本进行轮状病毒阳性鉴定,对鉴定为阳性的腹泻标本经处理后,采用多重一步反转录一聚合酶链反应（RT—PCR）对轮状病毒株的血清型进行鉴定,并通过对阳性标本的轮状病毒VP7基因的核苷酸序列进行分析、比对,以进一步鉴定毒株的血清型。结果对288例标本进行ELISA检测,RV阳性177例;血清型检测共测得G1型73例（41．2％）,G2型12例（6．7％）,G3型67例（37．9％）,另有25例为G1和G3混合型。结论轮状病毒是引起大庆地区婴幼儿病毒性腹泻的主要原因,对成人腹泻影响较小,2009～2011年大庆地区流行的RV以G,和G,型为主。此外,出现了G1和G3混合型,说明病毒血清型之间出现了一定重组变异。     

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.