外层致密纤维在Akap4基因缺陷致小鼠精子尾部多种形态异常中的作用

目的：探讨外层致密纤维(outer dense fobres，ODF)在Akap4基因缺陷引起小鼠精子尾部多种形态异常中 的作用。方法：通过基因编辑技术构建Akap4基因缺陷小鼠模型。实验分为2组：成年Akap4基因缺陷雄鼠为实验组 (n=7)，成年野生型雄鼠为对照组(n=7)，比较2组小鼠的体重和睾丸重量；采用计算机辅助精液分析(computer aided sperm analysis，CASA)检测精子活动力，改良巴氏染色(modifi ed pap staining)检测精子形态学，扫描及透射电镜观察精 子超微结构，免疫荧光检测精子尾部蛋白表达及定位，睾丸切片HE及PAS染色检测睾丸生精功能，透射电镜观察曲 精细管超微结构。结果： Akap4基因缺陷小鼠精子总活动力为8.81%，明显低于野生型小鼠(P<0.01)，且无正常形态精 子，尾部缩短与尾部卷曲比例达91.18%，明显高于野生型小鼠(P<0.01)，睾丸重量、睾丸生精功能、精子数量2组比 较差异无统计学意义(P>0.01)； Akap4基因缺陷精子的纤维鞘纵柱缺失，横肋柱结构部分残留，主段的3和8号ODF排 列紊乱，与ODF2蛋白定位结果相符；睾丸中精子纤维鞘发育障碍，无正常纤维鞘结构形成，但未见异常膨大的精子 尾部。结论：Akap4基因缺陷使纤维鞘横肋发育不良，纵肋的缺失引起3和8号ODF分离，“9+2”微管结构紊乱，使 主段管腔异常膨大，导致小鼠精子尾部多种形态异常。     

活动力不足精子尾部微管超微结构研究

以２２例活动力不足精子病人的精液为标本，对其精子进行形态学研究，以了解精子尾部的形态变化。电镜观察发现：１例精子尾部形态变化较一致。轴丝排列为９＋０微管形式；另１例精子尾部形态变化呈多样性，或部分周围微管和部分外围致密纤维以及中央微管消失，或所有微管和外周致密纤维排列混乱。结果显示部分活动力不足精子病人精子尾部的微管异常和Ｋａｒｔａｇｅｎｅｒ‘ｓ综合征特征相似，这种异常可能影响精子的运动能力，而导     

环磷酰胺对长爪沙鼠精子畸形的影响

目的 探讨环磷酰胺(CP)对长爪沙鼠精子畸形的影响.方法 取长爪沙鼠随机分成正常对照组,GP组(低、中、高剂量组即剂量分别为20 mg/kg、30 mg/kg、40 mg/kg),环磷酰胺经腹腔注射,连续注射5d,药后30d麻醉沙鼠,剖腹取出两侧附睾制备精子悬液涂片,用甲醇固定10 min,用2％的伊红染色30 min,蒸馏水洗片,显微镜观察精子形态.结果 低、中、高剂量组分别与正常对照组比较,长爪沙鼠精子畸形率差异极显著(P＜0.01);低、中剂量组分别与高剂量组相比较,长爪沙鼠精子畸形率差异极显著(P＜0.01);低剂量组与中剂量组比较,精子畸形率差异不显著(P＞0.05).形态学观察显示:环磷酰胺对长爪沙鼠精子影响畸形类型主要为尾折叠、尾粗细/长短/扭曲和无钩.结论 环磷酰胺对长爪沙鼠精子有一定的致畸率,且对长爪沙鼠精子尾部影响最大.在一定剂量内范围内,环磷酰胺对长爪沙鼠的精子致畸率无明显区别;当高于一定剂量,随着环磷酰胺用药剂量的增加,长爪沙鼠的精子致畸率也增加.     

长爪沙鼠生长繁殖性能的研究

目的　动物的生长繁殖性能是其生理学数据的重要组成部分 ,对实验动物的开发和应用具有指导意义。方法　选择离乳的长爪沙鼠 80只 (雌雄各半 ) ,一雌一雄长期同居。结果　每胎产仔 3～ 7只的居多 ,占总胎数的 77% ;平均每胎产仔 (5 .0 2± 2 .1 1 )只 ;每年产仔胎数 5～ 9胎的较多 ,占总胎数的 91 .7% ,平均 (7.4 9± 2 .0 1 )胎 /年 ;最早的初产周龄为 1 3周龄 ,最长的 5 2周龄 ,在 1 3～ 2 0和 2 5～ 2 8周龄的居多 ,占总对数的 80 %。长爪沙鼠的平均出生重为 3.5g ,成年鼠平均体重雌性 5 5 .6g ,雄性 6 7.2g。结论　普通级长爪沙鼠封闭群配种日龄最早在 6 5日龄 ,初产日龄超过 2 0 0日龄的长爪沙鼠应淘汰 ;性别对长爪沙鼠体重无显著影响     

长爪沙鼠T淋巴细胞参考值的初步探讨

用非特异性酸性α-乙酸萘酯酶(ANAE)法测定了30只健康的成龄长爪沙鼠循环血,其T淋巴细胞值为68.3±2.02,可初步作为长瓜沙鼠免疫功能指标,并提出ANAE反应物形态数量、大小等与细胞发育阶段关系的见解。     

长爪沙鼠IgG的纯化及抗血清的制备

目的纯化长爪沙鼠血清IgG,制备兔抗长爪沙鼠IgG抗血清。方法采用Hitrap Protein G亲和层析预装柱来纯化长爪沙鼠血清IgG;通过SDS-PAGE电泳和Western-Blotting免疫印迹法对长爪沙鼠血清IgG进行纯度鉴定,免疫兔子制备抗血清。结果7 mL长爪沙鼠血清纯化得到11 mg IgG;电泳和免疫印迹测定,IgG纯度大于95%;用纯化的IgG作抗原制备了兔抗血清,免疫双扩散测定效价达1∶32。结论建立了长爪沙鼠血清IgG的纯化方法,制备了长爪沙鼠IgG抗血清,证实长爪沙鼠血清IgG和Protein G具有较高的亲和性。     

长爪沙鼠发情周期规律与判定方法

目的 研究长爪沙鼠发情周期,揭示发情规律,优化判定方法。方法 连续18 d采集50只长爪沙鼠阴道上皮脱落细胞涂片,采用角化细胞计数法研究长爪沙鼠发情周期规律。比较瑞氏染色、HE染色和直接镜检判定发情周期4个时相的优缺点。结果 长爪沙鼠的发情周期有稳定型、不稳定型、假孕三种类型。其中稳定型占68.6%,发情周期为（106.3±35.0）h,可分为4个时相。4个时相角化细胞的比例分别为发情前期（13.5±7.8）%、发情期（86.7±9.9）%、发情后期（27.9±12.8）% 和发情间期（3.3±2.8）%。结论 角化细胞计数能准确地判定长爪沙鼠的发情周期及各个时相。直接镜检法能快速反映阴道脱落细胞的形态。     

蒙古长爪沙鼠染色体组型研究

目的　建立蒙古长爪沙鼠标准G带染色体的核型 ,提供可靠的细胞遗传学的背景数据。方法　雌、雄各 3只成熟长爪沙鼠 ,外周血淋巴细胞培养 ,制片、镜下观察分裂中期的淋巴细胞。随机计数 10 0个分裂细胞中的染色体数目 ,确立长爪沙鼠体细胞染色体数目。选择 10个典型细胞测量染色体基本数据 ,建立标准G带核型。结果　长爪沙鼠染色体数为二倍体 ,4 4条染色体 ,可划分A、B、C、D、E、F六组。中着丝点区域的 11对 ,主要分布A、C、E三组中 ;近中着丝点区域的染色体 5对 ,主要分布B组中 ;端着丝点区域的染色体 5对 ,主要分布D、F组中 ;x为第 6号中着丝点染色体 ,y为端着丝点的区域染色体大小在 14号与 15号染色体之间。结论　长爪沙鼠染色体为 2n =4 4 =2× 11m +2× 5sm +2× 5t+(x)m +(y)t     

二连浩特市2003年鼠间疫情特点分析

目的探讨在年降水量较多、植被恢复较好、鼠类种群数量达到一定程度时荒漠草原长爪沙鼠疫源地动物鼠疫疫情暴发流行的可能性。方法按照《内蒙古自治区鼠疫监测方案》及其实施细则进行,采用日弓形法和5 m夹线法。结果全年共调查样方76.75×105m2,平均鼠密度5.9只/×104m2,其中长爪沙鼠密度5只/×104m2;调查夜行鼠布夹1 000夹次,捕获率为7.3%;鼠体蚤染蚤率20.8%,平均指数为0.58;全年共剖验各类宿主动物12种697只,检出染疫长爪沙鼠32只,共培养各种蚤713组2 488匹,检出染疫蚤91组155匹,其余结果均为阴性。结论长爪沙鼠疫源地鼠密度异常增高时,动物鼠疫暴发流行的可能性极大,加强防控是避免波及到人间的重要手段。     

长爪沙鼠Willis环解剖学观察

自从Levine发现用夹闭长爪沙鼠双侧颈总动脉的方法能产生全脑缺血后,这种动物就开始被广泛应用于制备脑缺血模型,在国内外的研究中均有用这种动物制备脑缺血模型的报道。为了解长爪沙鼠Willis环的缺失程度,我们通过橡胶灌注法和单宁酸-氯化铁媒染法构筑了长爪沙鼠脑底动脉的结构,现报道如下：     

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.