重组蓝藻抗病毒蛋白N抗柯萨奇B3病毒的体外实验研究

目的:研究重组蓝藻抗病毒蛋白N (CV-N)体外抗柯萨奇病毒B组3型(CVB3)的作用. 方法:通过观察药物毒性、细胞病变效应(CPE)、MTT比色法检测细胞增殖,判断CV-N的细胞毒性及抗病毒效果. 结果:CV-N对Vero细胞毒性较低(TC50为359 μg/ml),对CVB3无直接灭活作用,对阻止CVB3病毒吸附细胞的作用优于病毒唑,而对病毒的复制无明显抑制作用. 结论:CV-N作用于CVB3病毒感染的早期,有可能作为柯萨奇病毒感染的预防药物.     

芪芩Ⅰ号对小鼠柯萨奇B3病毒性心肌炎的防治作用

目的:探讨复方制剂芪芩Ⅰ号(QiqinⅠ)水煎剂对柯萨奇病毒B3(CVB3)的抑制作用和对小鼠CVB3病毒性心肌炎的防治作用. 方法:①在Hep-2细胞上采用微量细胞病变抑制法,从直接灭活病毒、抑制细胞内病毒复制、阻断病毒吸附三方面研究芪芩Ⅰ号对CVB3的抑制作用. ②采用BALB/c鼠建立CVB3病毒性心肌炎的动物模型,建立病毒对照组和芪芩Ⅰ号组,然后在不同时间点处死小鼠,记录其心肌组织病变积分. 结果:①芪芩Ⅰ号水煎剂对细胞无毒且效应明显的原生药浓度是2.88 g/L和1.44 g/L. ②芪芩Ⅰ号可能具有直接灭活CVB3、抑制细胞内CVB3复制、阻断CVB3吸附细胞的作用. ③芪芩Ⅰ号组小鼠的心肌病变积分明显低于病毒对照组,二者比较有显著差异(P<0.01). 结论:体外实验证实芪芩Ⅰ号可能对CVB3有抑制作用,动物实验进一步证明芪芩Ⅰ号在体内对小鼠CVB3病毒性心肌炎有良好的防治作用.     

复方金钱草清热颗粒对柯萨奇病毒体外抑制作用的研究

目的：研究复方金钱草颗粒冲剂对柯萨奇B组病毒致人喉癌细胞（Hep-2）细胞病变是否有抑制作用。方法：①Hep-2细胞感染病毒的同时给药;②Hep-2感染病毒2h后再给药;③药物与病毒作用2h后再感染Hep2细胞。结果：复方金钱草清热颗粒对柯萨奇病毒段、B5致Hep-2细胞病变有抑制作用,同时对CBVs的抑制作用明显优于CBV3。结论：金钱草对柯萨奇病毒有体外抑制作用。     

芪芍五味子复方制剂抗CVB3病毒抑制心肌细胞凋亡机制的研究

目的 研究中药芪芍五味子复方制剂抗柯萨奇B3病毒(CVB3)效果及抑制病毒性心肌炎小鼠心肌细胞凋亡的机制.方法 采用细胞培养法观察细胞病变效应(CPE)与广谱抗病毒药物病毒唑对照,进行复方制剂体外抗病毒实验;BALB/c小鼠接种CVB3建立病毒性心肌炎动物模型,随机分为正常对照组、病毒对照组、复方制剂治疗组和维生素C与病毒唑联用组,观察各组心肌病理变化,Real-time定量PCR检测小鼠心肌CVB3RNA水平,流式细胞仪检测心肌细胞凋亡和坏死率.结果 复方制剂体外最大无毒浓度为19.53g/L,预防给药、同时给药和治疗给药3种方式细胞培养所需有效药物浓度均低于病毒唑;与同期病毒对照组小鼠比较,复方制剂治疗组小鼠心肌炎症性病理变化明显减轻,CVB3RNA拷贝数降低(P<0.05),心肌细胞凋亡和坏死率显著降低(P<0.05).结论 芪芍五味子复方制剂可有效抗病毒,抑制病毒诱导心肌细胞凋亡,对CVB3感染小鼠心肌有良好的保护作用.     

反义寡核苷酸抑制柯萨奇病毒B_3致病作用的实验研究

目的 :观察在细胞培养系统和小鼠体内反义寡核苷酸 (AODN)抑制柯萨奇病毒B3(CVB3)致病作用。方法 :观察细胞病变效应 (CPE) ,常规微量法滴定病毒的50 %组织细胞感染量 (TCID50)以及观察感染小鼠心肌组织的病理改变。结果 :1 AODN可推迟和减轻CVB3 感染细胞CPE ,且随AODN浓度增高 ,对CPE的抑制作用增强 ;培养上清中病毒滴度随AODN浓度升高而降低。2 随AODN剂量增加 ,感染小鼠心肌组织病毒滴度逐渐降低 ;心肌细胞病变程度逐渐减轻 ;心肌炎发病率也逐渐降低。结论 :AODN可能通过抑制CVB3 复制来保护Vero细胞和小鼠心肌组织抵抗感染。     

硝酸甘油和硝酸异山梨酯抗柯萨奇病毒的体外实验研究

目的研究硝酸甘油(GTN)和硝酸异山梨酯(ISDN)对柯萨奇B组3型病毒(CVB3)的体外抑制作用。方法用Hela细胞扩增并收获柯萨奇B组3型病毒(CVB3)后,测定CVB3的半数组织培养感染剂量(TCID50)和空斑形成单位(PFU),应用MTT法测定实验所用药物的细胞毒性,用细胞病变效应(CPE)抑制实验和空斑减少实验,观察和分析研究GTN、ISDN对CVB3的抑制作用。结果GTN、ISDN可以明显抑制病毒的细胞病变效应以及空斑形成(P<0.05)。结论GTN、ISDN等临床常用于抗心绞痛的硝酸酯类药物具有明确的抗CVB3感染Hela细胞的作用。     

黄芩茎叶活性部位抗病毒作用的研究

目的：研究黄苓茎叶活性部位（SLSBGAF）的抗病毒作用。方法：体外细胞培养观察1：2～1：256不同稀释度SLSBGAF对柯萨奇B族病毒（CoxB3、CoxB4、Cox B5）、流感病毒、副流感病毒、腺病毒（Adv3、Adv7）、呼吸道合胞病毒（RSV）．疱疹病毒（HSV-1、HSV-2）等10种病毒致细胞病变（CPE）的抑制作用；流感病毒感染小鼠模型，分别观察SLSBGAF对小鼠肺指数、间接荧光染色观察肺内病毒增殖的影响，并以小鼠存活时间和死亡率为指标观察药物的保护作用。结果：SISBGAF对CoxB3等10种病毒所致细胞病变有明显的抑制作用，有效浓度为0．20～1．56mg／mL，IC50值为0．08～0．34mg／mL，治疗指数（TI）值分别为4和8；sLSBGAF（54．3，108．5，217mg／kg）灌胃给药对流感病毒引起的小鼠肺炎有治疗作用，肺指数和病毒的肺内增殖低于对照组，并呈一定的量效相关性；此外对流感病毒所致小鼠死亡有不同程度的保护作用，217mg／kg剂量组作用明显，死亡保护率为40％，生命延长率为39．08％。结论：SLSBGAF可用于流感病毒、副流感病毒、柯萨奇病毒等相关感染性疾病的预防和治疗。     

小柴胡汤分解剂抗柯萨奇B3病毒感染及其对心肌炎防治作用的研究

目的 :探讨小柴胡汤分解剂在体外及体内抗柯萨奇 B3病毒 (CVB3)及保护心肌细胞的作用。方法 :用小柴胡汤分解剂通过体外微量细胞培养 ,作用于被 CVB3直接损伤的细胞模型及经腹腔感染 CVB3m Balb/ c小鼠心肌炎模型 ,分析不同浓度分解剂的效应。结果 :在体外细胞上分解剂 A、B液自 1∶ 32稀释度对 vero细胞的毒性作用已丧失 ,至 1∶ 2 5 6稀释度时 ,仍能对抗 CVB3对细胞致病变作用。在体内分解剂 A、B液治疗组小鼠 35只 ,心肌炎发生率 37.14% ;而 2 0只对照组小鼠心肌炎发生率 5 9.1% ,两者有显著性差异 (χ2 =4.6 8,P <0 .0 5 ) ,在预防给药组更为明显 (P <0 .0 1)。结论 :小柴胡汤分解剂 A、B液在体内、体外均具有抗病毒及保护心肌的作用。     

4-苯乙烯磺酸马来酸酐共聚物抑制HSV-2感染研究

目的:探索4-苯乙烯磺酸马来酸酐共聚物(PSM)对单纯疱疹病毒Ⅱ型(HSV-2)感染细胞或小鼠的抑制作用。方法:建立Vero-ICP10细胞系,通过Luciferase检测不同浓度PSM对感染细胞中HSV-2的抑制作用,并通过In cell western技术检测HSV-2 gD蛋白表达,确定不同PSM浓度对HSV-2 gD蛋白表达的抑制作用;建立HSV-2阴道感染小鼠模型,观察PSM对HSV-2经生殖道感染小鼠的抑制作用。结果:Luciferase实验和In cell western实验结果均表明PSM具有抗HSV-2感染细胞的作用;小鼠模型中,5%的PSM能有效防止小鼠感染HSV-2(P<0.001)。结论:PSM在体内和体外模型中均可以抑制HSV-2感染。     

玉丹荣心丸抗急性病毒性心肌炎的实验研究

目的观察中成药玉丹荣心丸对B组柯萨奇病毒(CVB)感染Balb/c小鼠所致急性病毒性心肌炎的治疗效果的研究。方法Balb/c鼠腹腔接种1000 TCID50/0.1ml CVB,经过两周的持续注射成功获得急性病毒性心肌炎的动物模型。将动物随机分为病毒对照组、药物组,药物组使用中成药玉丹荣心丸进行灌胃给药,而病毒对照组不使用任何药物,28天后观察小鼠心肌组织的病理改变。结果药物组小鼠心肌组织的病理改变轻微,并有好转迹象,而病毒对照组病理改变严重,有死亡。结论玉丹荣心丸对B组柯萨奇病毒感染所致的急性病毒性心肌炎有很强的治疗效果。     

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.