青岛地区2000～2003年肾综合征出血热的病毒基因分型

①目的 调查山东省青岛地区2000～2003年汉坦病毒的流行情况，研究病毒在该地区的分子流行病学特征。②方法 采集肾综合征出血热(HFRS)急性期病人血清标本64份，提取血清中病毒RNA作为模板，根据GenBank中汉坦病毒基因序列，设计HTN型和SEO型的通用外套引物，同时分别设计HTN型特异性引物和SEO型的特异性引物作为内套引物，RT—nest—PCR扩增汉坦病毒基因组M片段G1区基因序列并测序。测序结果利用DNA Star软件进行核苷酸序列分析。③结果 在64例标本中用HTN型特异性引物扩增阳性6例，占9％；用SEO型特异性引物扩增阳性25例，占39％；总阳性率为48％。总体来看青岛地区流行的汉坦毒株以SEO型为主。SEO型汉坦病毒间核苷酸序列的差异相对较小，在核苷酸序列水平其基因的离散度为0．3％～8．9％。HTN型汉坦病毒的变异率较高，其基因的离散度为2．6％～11．2％。④结论 青岛地区汉坦病毒的流行以SEO型为主，HTN型并存，其中HTN型主要发生在胶南市。     

山东地区汉坦病理重要基因序列进化关系的研究

目的：研究山东地区汉坦病毒的基因型分布。方法：应用RT－PCR法和直接核酸测序法，对来源于山东省11个肾综合征出血热流行地区的患者中获得的31份汉坦病毒部分M基因片段进行了扩增和测序。结果：核苷酸序列的同源性及系统发生树分析表明，山东地区SEO型汉坦病毒至少存在4个不同亚型，HTN型汉坦病毒至少发现有4个分支，不同HTN型汉坦病毒亚型间的核苷酸序列的差异达13．0％－22．7％，而不同SEO型汉坦病毒分支间相对保守，差异仅5．0％－7．7％。结论：山东地区存在汉坦病毒的不同的基因亚型，这一发现将促进汉坦病毒流行规律、致病性和疫苗的研究。     

肾综合征出血热汉坦病毒分型及序列特征的研究

目的 分析黑龙江省肾综合征出血热（HFRS）患者携带汉坦病毒的基因型别及序列特征.方法 采集临床诊断为肾综合征出血热病人的全血60例,其中通过金标法检测出血热特异性抗体阳性40例、阴性20例,血凝块提取汉坦病毒RNA,应用RT-Nest-PCR及核苷酸序列测定分型技术对黑龙江地区的汉坦病毒进行分型研究,设计汉坦病毒M片段通用引物（MOF,MOR）及HTNV和SEOV的M片段特异性引物（HTNMF、HTNMR,SEOMF、SEOMR）,应用Nest-PCR进行扩增,回收阳性扩增产物进行基因序列测定,将所测基因序列与国内外HV病毒株序列进行核苷酸同源性分析,构建M基因种系进化树.结果 出血热特异性抗体阳性的病例中扩增出HTN型19份,抗体阴性的病例中扩增出HTN型1份、SEO型l份.总体来看黑龙江省流行的汉坦病毒毒株主要为HTN型和SEO型,以HTN型为主.经核苷酸及氨基酸同源性和种系进化分析表明其黑龙江地区的病毒株同源性较高,其中HTN型与76 ～ 118株同源性较高,SEO型与Z37株的同源性较高.结论 黑龙江省的HV感染病例多为HTN型,与其他GenBank中的各地标准株的核苷酸及推导的氨基酸序列均有一定差异,在系统进化树中分析HTN型与76-118株较为接近.SEO型与国内Z37较为接近.结合临床症状发现其临床症状较为相似的在系统进化发生树中较为接近.     

山东地区汉坦病毒重要基因序列进化关系的研究

目的 :研究山东地区汉坦病毒的基因型分布。方法 :应用RT -PCR法和直接核酸测序法 ,对来源于山东省 11个肾综合征出血热流行地区的患者中获得的 31份汉坦病毒部分M基因片段进行了扩增和测序。结果 :核苷酸序列的同源性及系统发生树分析表明 ,山东地区SEO型汉坦病毒至少存在 4个不同亚型 ,HTN型汉坦病毒至少发现有 4个分支 ,不同HTN型汉坦病毒亚型间的核苷酸序列的差异达 13.0 %～2 2 .7% ,而不同SEO型汉坦病毒分支间相对保守 ,差异仅 5 .0 %～ 7.7%。结论 :山东地区存在汉坦病毒的不同的基因亚型 ,这一发现将促进汉坦病毒流行规律、致病性和疫苗的研究     

河北省肾综合征出血热主要流行区鼠携带汉坦病毒基因序列分析

目的了解河北省肾综合征出血热（haemorrhagic fever with renal syndrome,HFRS）主要流行区汉坦病毒主要流行株的基因型及其变异情况。方法收集2012年河北省东北部地区捕获的啮齿动物,均为褐家鼠,取鼠肺组织,用间接免疫荧光法（indirect immunofluorescence assay,IFA）检测,抗原阳性的标本进行汉坦病毒（Hantavirus,HV）M部分片段扩增并测序,与该地区以往序列及其他部分标准株进行系统发生树及同源性分析。结果收集164份鼠肺标本,经IFA检测26份阳性,挑选9份有代表意义的标本进行逆转录巢式PCR扩增,9份鼠肺标本经RT-PCR检测均为汉城（SEO）型HV,病毒间变异不大,同源性达95.2%~100.0%。以往测序株与新测序株比较同源性为99.0%~100.0%。系统发生树分析结果表明9份标本均为SEO型HV,且为S3亚型。结论河北省HFRS主要流行区HV以SEO型为主,亚型为S3亚型。同型HV基因相对保守,具有区域稳定性特征。     

黑龙江省1株汉滩型汉坦病毒的分离及基因鉴定

目的从鼠肺中获得汉坦病毒分离株并进行基因鉴定与分型。方法用组织培养法分离病毒，并进行RT—PCR鉴定，之后对目的片段进行序列测定和序列分析。结果从黑线姬鼠鼠肺中分离到一株汉坦病毒，编号HL83。用汉滩型（HTN）汉坦病毒M基因特异性引物检测得到预计大小的片段，此片段的核苷酸序列分析表明，HL83与HTN型参考毒株的同源性较高，与汉城型（SEO）参考毒株的同源性较低，进化树分析表明HL83位于HTN型所在的支系。结论HL83为HTN型汉坦病毒。     

湖北省不同年份汉坦病毒的RT-PCR检测及分型

目的：探讨湖北省不同年份引起肾综合征出血热的汉坦病毒的主要型别，为进一步研究该型汉坦病毒的基因变异打下基础。方法：根据汉坦病毒M基因片段G1编码区核苷酸序列设计I型和Ⅱ型分型引物，利用RT—PCR对105份不同年份的病程在7日以内的肾综合征出血热(HFRS)患(经临床诊断和ELISA确诊)血清标本进行检测及基因分型，并对扩增出的基因片段选用3种限制性内切酶AluI、RsaI、MboI进行酶切分析。结果：用I型引物通过：RT—PCR检测75份血清标本(1996～2000年)和30份血清标本(1986～1987年)的阳性率分别为77．3％和10％，而用Ⅱ型引物检测前阳性率为8．0％，后均为阴性。对4份I型引物扩增产物用3种限制性内切酶进行酶切分析，发现4份阳性产物其限制性片段长度多态性均与I型汉坦病毒标准株76～118完全不同，但其中3份与I型汉坦病毒湖北地方株HV114相同，1份与HV114不完全相同。结论：不同年份引起湖北地区HFRS的主要毒株均为I型汉坦病毒(HTN)湖北地方株。     

上海地区汉坦病毒基因分型和序列分析

目的 研究上海地区汉坦病毒基因型分布。方法 采用逆转录聚合酶链反应(RT-PCR)、核苷酸测序法和系统发生树分析上海宿主动物中获得的36株汉坦病毒M基因片段。结果 上海地区郊区和郊县汉坦病毒流行株有HTN型和SEO型,以HTN型为主;市区流行株为SEO型。核苷酸序列同源性及系统发生树分析表明上海地区在HTN型汉坦病毒存在3个不同分支,不同HTN型病毒间的核苷酸的差异在6％～19％。结论 上海地区汉坦病毒以HTN型为主。可能存在汉坦病毒的不同基因亚型,对本市肾综合征出血热的防治有重要指导意义。     

汉坦病毒湖北株M片段G1编码区基因变异研究

目的 :探讨近年来汉坦病毒HTN湖北株M基因片段G1编码区部分基因序列的变异情况。方法 :根据汉坦HTNM片段G1编码区核苷酸序列设计引物 ,利用RT PCR方法对 75份病程在 7d以内的肾综合症出血热 (HFRS)患者 (经临床诊断和ELISA确诊 )血清标本进行检测 ,并对扩增出的目的基因片段选用 3种限制性内切酶AluⅠ、RsaⅠ、MboⅠ进行酶切 ,根据限制性片段长度多态性分析筛选出变异毒株 ,继而通过序列测定 ,得到变异毒株G1编码区部分核苷酸序列 ,并与标准株HTN76 118及地方株HTN 114的相应核苷酸序列及氨基酸序列进行比较分析。结果 :75份血清标本通过RT PCR方法进行检测 ,阳性率为 77.3 %。扩增片段用 3种限制性内切酶AluⅠ、RsaⅠ、MboⅠ进行酶切分析 ,发现 4份阳性产物其限制性片段长度多态性与HTN 76 118均不相同 ,但其中 3份阳性产物限制性片段长度多态性与地方株HTN 114基本相同 ,1份与HTN 114株不完全相同。对该扩增产物的核苷酸序列和氨基酸序列进行同源性分析表明 ,其核苷酸序列与标准株 76 118的同源性为 83.3 % ,与地方株HTN 114的同源性为 96 % ;推导氨基酸序列与HTN 76 118的同源性为 93.2 % ,与地方株HTN 114的同源性为 99%。结论 :①近年来引起湖北地区HFRS的主要毒株仍为HTN地方株。②从分析的病     

巴彦淖尔市肾综合征出血热疫情分析

目的调查1999年后巴彦淖尔市及周围区域流行性出血热疫情呈逐年上升趋势的原因。方法对2005年春末在巴彦淖尔市临河区有HFRS病人的居民区捕获褐家鼠119只、小家鼠21只、获得鼠血条126份、9份病人血清和2份正常人血清样本。以间接免疫荧光和RT—PCR等流行病学检测技术，综合全市人文地理特征分析。结果恢复期病人及病人密切接触者的IsG均显示对SEO型L99株抗原的反应滴度高于HTN型76-118株，从褐家鼠筛选出9个阳性标本，对其部分S和部分M核苷酸序列进行系统发生分析，均为SEOI亚型。结论汉城病毒为巴彦淖尔市流行性出血热暴发的主要病原体。褐家鼠为优势鼠种，有很高的感染率和带病毒率，是主要传染源和储存宿主。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.