出生后大鼠螺旋神经节神经元损伤的病理变化

目的 证实出生后第2周大鼠螺旋神经节神经元对耳毒性药物敏感。方法 选用出生后7d SD大鼠36只，连续14d肌注新霉素，每天150mg／kg，分别于停药后1、7、14、28d、3月和6月各处死6只。取平行蜗轴的超薄切片，透射电镜下观察底回、中回螺旋神经节神经元。结果 大鼠螺旋神经节神经元变性继发于毛细胞和支持结构的损伤；停药1d组存在细胞凋亡现象；损伤早期神经元发育延迟，损伤晚期神经元由成熟向未分化型改变。结论 出生后第2周是大鼠螺旋神经节神经元发育的重要阶段，此阶段对药物的敏感性高。     

神经生长因子在新生大鼠耳蜗的定位及其意义

目的 观察神经生长因子在新生大鼠耳蜗中的定位,为进一步研究其在听觉通路中的作用机制提供依据.方法 出生后5～7 d健康SD大鼠20只,制备耳蜗石蜡切片,用兔抗小鼠神经生长因子(nerve growth factor,NGF)多克隆抗体行免疫组织化学染色(S-P法),检测NGF在耳蜗中的表达.结果 新生SD大鼠膜迷路呈类三角形结构,可清晰看见幼稚柯替器、螺旋神经节细胞、蜗轴中的细胞染色阳性,细胞核及细胞质内均有表达.结论 NGF对出生后5～7 d大鼠螺旋神经节细胞和毛细胞的存活、成熟密切相关.     

新生大鼠耳蜗螺旋神经节神经元的原代培养

[目的]建立原代细胞培养大鼠耳蜗螺旋神经节神经元(SGN)的方法.[方法]选择出生后5d的SD大鼠的SGN进行原代细胞培养,采用免疫细胞染色法进行细胞学鉴定.[结果]新生SD大鼠SGN在原代培养条件下获得良好的细胞形态及表型分化,表现出稳定的神经元可塑性.[结论]原代细胞培养方法可获得状态良好的SGN细胞.     

出生后大鼠耳蜗不同发育阶段connexin26蛋白的表达

目的:研究缝隙连接蛋白26(cx26)在出生后大鼠耳蜗不同发育阶段的分布.方法:采用免疫组织化学SP法检测cx26在36只大鼠(1周龄组、3周龄组和9周龄组各12只)耳蜗的表达.结果:cx26在3组均有较强的表达,阳性表达主要分布于耳蜗外侧壁血管纹和螺旋韧带,Corti氏器中的支持细胞间以及毛细胞和支持细胞间,螺旋缘以及螺旋神经节细胞.cx26在大鼠耳蜗外侧壁、Corti氏器与螺旋神经节的阳性细胞平均积分光密度3组间两两比较,差异均无统计学意义(P均>0.05).结论:大鼠耳蜗发育过程中cx26的表达趋于稳定,没有随着听力的发展而变化.     

大鼠下丘脑弓状核神经元的衰老性变化——光,电镜定量研究

目的：为了研究中药的抗衰老作用，为弓状核提供形态学资料；方法：采用光、电镜技术和形态计量方法对20只SD老年大鼠下丘脑弓状核神经元进行了形态定量研究。结果：①弓状核由亮、暗神经元胞体，胶质细胞胞体，血管和神经毡组成；②老年大鼠弓状核神经元胞体的面积分数、面数密度和胞体、胞核的等效直径未见明显改变（P＞0．05）；③老年大鼠弓状核亮细胞内细胞器和分泌颗粒的面积分数和面数密度未见改变（P＞0．05），而暗细胞内线粒体、溶酶体的面积分数和面数密度均增加（P＜0．05），粗面内质网、高尔基复合体、游离核糖体和分泌颗粒的面积分数和面数密度明显减少（P＜0．05）。结论：①老年大鼠弓状核内亮、暗细胞的形态定量改变不一致，呈现不同的反应；②衰老引起神经内分泌功能紊乱可能与弓状核的衰老变化直接相关。     

EGCG对阿米卡星诱导大鼠螺旋神经元凋亡的保护作用

目的 探讨表没食子儿茶素没食子酸酯(EGCG).对阿米卡星诱导的大鼠耳蜗螺旋神经元凋亡是否具有保护作用.方法 设生理盐水对照组、阿米卡星(450 mg·kg-1·d-1共14 d)损伤组和阿米卡星加EGCG(50 mg/kg·kg-1·d-1共14 d)保护组.各组动物均深度麻醉后半身灌注固定处死,取右侧耳蜗标本固定、脱钙、石蜡轴位包埋、轴位切片,近中轴位切片行末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记(TUNEL)免疫组化染色,光学显微镜下检测耳蜗螺旋神经节细胞表达情况、并通过切片图像分析进行半定量处理比较.结果 正常组螺旋神经元胞浆及细胞核基本上均不着色,损伤组部分螺旋神经元胞核着色,保护组亦少部分螺旋神经元胞核着色,图像分析结果显示损伤组与对照组、损伤组与保护组之间的染色阳性率差异有统计学意义(P＜0.01).结论 EGCG可以减轻阿米卡星诱导大鼠耳蜗螺旋神经元凋亡.     

大鼠翼腭神经节、耳神经节和脑底动脉壁神经纤维一氧化氮合酶表达的年龄变化

目的 :观察大鼠翼腭神经节、耳神经节及脑底动脉壁神经纤维一氧化氮合酶表达的年龄变化规律。方法 :用还原型尼克酰胺腺嘌呤二核苷酸磷酸脱氢酶 (NADPH -d)组织化学法对不同年龄组翼腭神经节、耳神经节及脑底动脉壁神经纤维一氧化氮合酶表达进行观察。结果 :NOS神经细胞在翼腭神经节、耳神经节中总体呈均匀散在分布 ,从幼年到成年 ,密度呈逐渐下降趋势 ,而NOS神经元胞体大小逐渐增加 ,核浆比逐渐下降 ,至成年时最大 ,直至老年无明显改变。脑底动脉神经纤维密度从幼年到成年逐渐增高 ,从成年起维持较高密度直到老年。结论 :出生后翼腭神经节和耳神经节及脑底动脉壁神经纤维一氧化氮合酶表达的年龄变化有其特定的规律     

稳定表达神经营养素-3的NIH3T3细胞建立及鉴定

目的：建立稳定表达NT3的NIH3T3细胞株，通过体外共培养实验观察其对耳蜗螺旋神经节细胞生长的影响．方法：以阳离子脂质体作为载体，将携带有外源性基因NT3的重组真核表达载体pIRES2-EGFP-NT3转染小鼠成纤维细胞，进行RT-PCR，Western-blot分析及免疫组化鉴定，并与耳蜗螺旋神经节细胞进行共培养实验．结果：得到抗G-418的阳性细胞克隆；RT-PCR的产物约为744 bp；Western-blot可见一清晰的蛋白条带，与NT3的蛋白分子量相符合．NT3免疫组化染色结果显示转染NT3-cDNA的NIH3T3细胞胞质呈棕黄色着色．NT3转染组细胞与耳蜗螺旋神经节细胞共培养2wk后，与对照组相比，耳蜗螺旋神经节细胞无明显减少．结论：成功建立了稳定表达NT3的NIH3T3细胞株；该细胞对耳蜗螺旋神经节细胞有很好的营养支持作用．     

大鼠弓状核神经元的昼夜节律变化──光、电镜定量研究

目的：探讨弓状核的昼夜节律变化。方法：采用光、电镜技术和体视学方法对40只SD大鼠弓状核神经元的昼夜节律变化进行了形态定量研究。结果：①四组大鼠弓状核神经元的面积分数、面数密度和神经元胞体、胞核的等效直径未见明显改变（P＞0．05）;②从Ⅰ组至Ⅲ组,亮、暗细胞内细胞器逐渐增加,从Ⅲ组到Ⅳ组逐渐下降。③在Ⅲ组,弓状核内亮、暗细胞的粗面内质网、高尔基复合体、游离核糖体和分泌颗粒的面积分数和面数密度显著增高（P＜0．05）。结论：提示大鼠弓状核神经元细胞器发生昼夜节律变化,这种变化可能与视交叉上核的节律变化有关。     

谷氨酸对豚鼠耳蜗传入神经元的兴奋毒性损害

目的：观察不同剂量谷氨酸导致豚鼠耳蜗传入神经元兴奋性损伤性质及耳蜗形态、电生理改变。方法：经耳蜗鼓阶灌注高浓度谷氨酸(Glu-H,20 mmol/L,10 μl)、低浓度谷氨酸(Glu-L,10 mmol/L,10 μl)后检测不同时点复合动作电位 (CAP)反应阈,并观察耳蜗显微和超微结构变化。 结果：①Glu-L组(≤1 d)及Glu-H组CAP反应阈与正常对照组比较明显提高（P<0.001）;Glu-L组（≥1周）CAP反应阈与正常对照组比较差异无统计学意义（P>0.05）。②两给药组（Glu-H,Glu-L）蜗轴半薄切片Ⅰ型螺旋神经节细胞部分出现空泡样变化,胞浆变淡,胞核皱缩;正常对照组与给药组（Glu-H）之间螺旋神经节细胞病变率比较差异有统计学意义(P<0.05)。③两给药组电镜下耳蜗传入神经树突末梢、有髓神经纤维、Ⅰ型螺旋神经节细胞有变性坏死,呈明显的剂量依赖性,Glu-L组的神经元病变4周内可基本恢复,而Glu-H组进行性加重,由变性发展到坏死。结论：谷氨酸鼓阶灌注可导致豚鼠耳蜗传入神经元形态和功能的损害。小剂量（10 mmol/L）的谷氨酸可导致耳蜗传入神经系统的可逆性损害,而大剂量(20 mmol/L)的则可导致不可逆的神经元损害。     

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.