实验动物大、小鼠冠状病毒的血清学调查与分析

实验动物是冠状病毒的重要自然宿主,具有较高的感染率。它可以引起小鼠的病毒性肝炎(MHV),大鼠冠状病毒病(RCV)感染,大鼠涎泪腺炎,猫传染性腹膜炎,还能引起犬和兔的感染。因此,调查目前实验动物的冠状病毒感染情况,对探究SARS病的起源和开展SARS病的防治以及模型研究是十分必要的。1　材料和方法11　大小鼠血清的采集　用摘眼球采血方法对各有关单位的实验大、小鼠进行采血,分离血清并置-20℃以下冰箱保存待检。12　MHV和RCV抗体ELISA检测试剂盒　购自中国药品生物制品鉴定所,该试剂盒由抗原包被板、稀释液、阴性血清对照、阳性血…     

湖南省2004～2006年实验动物病毒学质量监测结果分析

目的了解湖南省实验动物病毒学质量。方法按GB14922.2-2001和GB14926-2001进行,在相关单位生产繁殖群以单纯随机抽样原则采样检测。结果实验小鼠MHV和Sendai病毒出现抗体阳性并出现疫情;普通级兔出血症项目不合格;实验大鼠未检出相关病毒。结论实验动物病毒学质量存在较大问题,应加强监测。     

IVC换笼标准操作程序与小鼠病毒病原的传播风险

目的研究独立通风笼具换笼标准操作程序与实验动物负压屏障设施内使用IVC(individual ventilated cage,IVC)设备饲养小鼠的病毒感染风险。方法选择小鼠易感的小鼠肝炎病毒(MHV) A59株感染6周龄BALB/c雄性小鼠,按照IVC换笼标准操作程序进行换笼操作;使用IFA和ELISA血清学方法以及针对MHV的核酸检测方法测定同一笼架具相邻笼盒、同一IVC系统不同笼架具的不同位置、以及IVC系统外负压屏障设施内哨兵动物的MHV感染情况。结果接种病毒小鼠在感染6 d后出现临床症状,粪便中可检出MHV病原;动物感染3周后可检出MHV抗体阳性;其他监测组在5周实验终点均无MHV阳性检出。结论 IVC换笼标准操作程序可指导IVC换笼操作,用以控制病毒类病原对小鼠的感染风险。     

山东省实验动物质量监督检测现状及对策

目的 探讨目前山东省实验动物质量监督检测的现状及对策.方法 通过对近年来山东省实验动物质量监督检测情况,包括实验室建设,大、小鼠、SPF鸡(鸡胚)质量监督检测结果,以及检测过程中存在的困难进行统计、分析和总结,在肯定取得的成绩的同时,找出存在的不足并探讨解决办法.结果 2009～2011年检测的SPF级和清洁级大、小鼠中病原菌检出率很低,仅检出铜绿假单胞菌、肺炎克雷伯杆菌和沙门氏菌各一例;病毒检出主要集中在MHV、SV、RCV、PVM、MVM、TEMV;体外寄生虫、鞭毛虫和蠕虫都有检出,而鞭毛虫检出率最高.2010～2011年SPF鸡(鸡胚)质量监督检测结果统计表明,病毒检出主要集中在REO、CAV、AEV、EDS等,尤其以REO检出率最高.结论 经过近几年的建设,山东省实验动物质量监督检测体系已逐步走向完善;但监督检测过程中仍然存在一些问题,比如抽检比例相对较低;受检测试剂的限制,检测方法比较单一等.     

实验动物微生物、寄生虫抽样调查及分析

目的 监控实验动物微生物、寄生虫质量.方法 细菌检测:常规培养、生化鉴定,免疫荧光试验(IFA)查抗体.病毒检测:酶联免疫吸附试验(ELISA)、免疫酶试验(IEA)、免疫荧光试验(IFA)、血凝试验(HA)、血凝抑制试验(HAI)方法检测抗体.真菌检测:常规培养、镜检.寄生虫检测:涂片镜检、间接血凝试验(IHA)方法查抗体.结果 五年来,SPF级动物检出了绿脓杆菌、嗜肺巴斯德菌、金黄色葡萄球菌、鞭毛虫,另外,泰泽病原体、小鼠细小病毒(MVM),小鼠仙台病毒(M-SC)抗体阳性.清洁级动物检出了蠕虫,且泰泽病原体、大鼠仙台病毒(R-SC)、小鼠仙台病毒(M-SV)、小鼠肝炎病毒(MHV)抗体阳性.普通级动物检出了志贺菌、皮肤病原真菌、体外寄生虫、弓形虫,同时检出兔出血症病毒(RHDV)、犬细小病毒(CPV)、犬瘟热病毒(CDV)、犬肝炎病毒(ICHV)未达抗体保护效价,犬布鲁杆菌、猕猴疱疹病毒Ⅰ型(BV)抗体阳性.不同生产企业动物质量差异很大,有的动物合格率达100%,有的则同一动物有多种病原感染.结论 实验动物微生物、寄生虫质量控制有待加强.     

上海地区实验动物病原体感染指数分析

目的 为了更直观的了解实验动物质量控制情况,本文引“感染指数”这一评估指标。探索测算感染指数的最佳方法,以便更加科学的反映实验动物的感染状况。方法 感染指数,也称感染度,它是对实验动物质量监测的定性指标。通过加工汇总实验动物的病原体感染率,综合反映某一特定实验动物群体的病原体感染状态或趋势的一种指标。结果 总体上,小鼠病原体感染指数呈逐年下降的趋势,而大鼠的病原体感染指数则逐年上升。分别比较各类病原体感染指数发现,小鼠中寄生虫感染指数最高,而大鼠中细菌感染指数最高。结论 感染指数分析揭示了实验动物质量的基本状况,需要加强对小鼠寄生虫的监测,而大鼠的细菌检测需要投入更多关注。小鼠病原体感染情况得到控制,大鼠各类病原体的控制情况均逐年趋于严重。     

长爪沙鼠小鼠肝炎病毒(MHV)RT-PCR检测方法的建立及初步应用

目的建立长爪沙鼠小鼠肝炎病毒(MHV)RT-PCR检测方法,应用于长爪沙鼠、小鼠等实验动物MHV的检测。方法根据已发表的小鼠肝炎病毒(MHV)S基因序列,设计合成引物。提取MHV细胞毒RNA,以其为模板,进行PCR扩增。优化反应条件,进行特异性、敏感性、稳定性、重复性试验。并对65只长爪沙鼠及12只小鼠进行检测。结果建立的MHV RT-PCR检测方法特异、敏感、稳定。以MHV RNA逆转录产物为模板,所能检测RNA最小模板浓度为3.1 pg/μL,可检测病毒最小滴度为10-3/mL。65只沙鼠经RT-PCR检测,均为阴性,12只小鼠经RT-PCR检测,有3只MHV阳性,测序结果与Genbank中MHV核酸序列同源性均为97%。结论建立的长爪沙鼠小鼠肝炎病毒(MHV)RT-PCR检测方法可用于长爪沙鼠、小鼠等实验动物MHV的检测。     

长爪沙鼠小鼠肝炎病毒（MHV）RT-PCR检测方法

目的 建立长爪沙鼠小鼠肝炎病毒（MHV）RT-PCR检测方法,应用于长爪沙鼠、小鼠等实验动物MHV的检测。方法 根据已发表的小鼠肝炎病毒（MHV）S基因序列,设计合成引物。提取MHV细胞毒RNA,以其为模板,进行PCR扩增。优化反应条件,进行特异性、敏感性、稳定性、重复性试验。并对65只长爪沙鼠及12只小鼠进行检测。结果 建立的MHV RT-PCR检测方法特异、敏感、稳定。以MHV RNA逆转录产物为模板,所能检测RNA最小模板浓度为3.1pg/μl,可检测病毒最小滴度为10-3.ml-1。65只沙鼠经RT-PCR检测,均为阴性,12只小鼠经RT-PCR检测,有3只MHV阳性,测序结果与GenBank中MHV核酸序列同源性均为97%。结论 建立的长爪沙鼠小鼠肝炎病毒（MHV）RT-PCR检测方法可用于长爪沙鼠、小鼠等实验动物MHV的检测。     

在生命科学研究中影响动物实验结果的多因素分析

动物实验是生命科学研究最主要的方法之一，作者对影响动物实验结果的实验动物因素、环境因素、营养因素、化学药剂、隔离和饲养密度、运输、抓取和保定、微生物因素、社会因素和人类因素等进行了分析，为保障动物实验结果的准确性提供了依据。     

山东省实验动物寄生虫学质量检测结果

1993～ 2 0 0 0年间作者按国家标准对山东省 38个单位 10 6个普通级实验动物群、7个清洁级实验动物群进行了寄生虫学检测。结果表明 :普通级实验动物中阳性率 32 .93% (30 3/ 92 0 ) ,清洁级实验动物中阳性率 2 3.33% (14 / 6 0 )。影响普通级实验动物达标的主要是体外寄生虫 ,而肠道蠕虫依然是亚屏障系统中实验动物寄生虫感染的主要因素山东省实验动物寄生虫学质量检测结果@路莉$山东省实验动物中心!济南250002 @康爱丽$山东省实验动物中心!济南250002实验动物;;寄生虫;;检测…     

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.