促性腺素释放激素拟似剂对大鼠生长的影响及机制

目的　观察抗早熟药物-促性腺素释放激素拟似剂(GnRHa)对大鼠身长增长的影响,并探讨药物作用的机制。方法　选择正常青春启动期雌性SD大鼠60只(4 周龄),随机分为 GnRHa注射组和空白对照组,每周观测身长、体重,观察阴门开启情况;用逆转录和荧光定量 PCR的方法,从 mRNA转录水平分析各组动物下丘脑 GH IGF轴相关激素及其受体表达水平的变化。结果　(1) 空白组大鼠阴门至 6 周龄全部开启,而 GnRHa组至 10 周龄全部开启;GnRHa组青春期生长速度较空白组减慢,但两组成年期身长无显著差异;(2) 用药期间,两组均未影响下丘脑 GH IGF轴中促生长激素释放激素及其受体(GHRH、GHRHR)的 mRNA转录水平,但 GnRHa 组生长抑素(SRIH) mRNA转录水平增高;生长激素(GH)及其受体(GHR)、胰岛素样生长因子1(IGF-1)及其受体(IGF-1R)mRNA转录水平降低。结论　(1) GnRHa可延缓大鼠性成熟发育,同时减慢大鼠青春期生长速度,但对成年期身长无影响;(2) 其作用主要通过影响下丘脑 GH IGF轴中SRIH、GH、GHR、IGF-1和 IGF-1R而实现。     

促性腺激素释放激素拟似剂对青春期大鼠生长轴与性腺轴相关基因表达的影响

目的 探讨促性腺激素释放激素拟似剂(GnRHa)对青春期大鼠生长轴与性腺轴的影响及其作用机制。方法 青春期大鼠应用GnRHa后，取其下丘脑、垂体、卵巢、下肢骺软骨等组织，应用荧光定量PCR(FQ-PCR)技术检测组织中相应激素的基因表达水平。结果 GnRHa可使下丘脑中促性腺激素释放激素(GnRH)和垂体中GnRH受体(GnRHR)基因表达水平下降，下丘脑中生长激素释放抑制激素(SRIH)基因表达水平增高，生长激素释放激素(GHRH)基因表达无变化；垂体中生长激素(GH)、卵巢中雌激素受体(ER)、下肢骺软骨中胰岛素样生长因子—1(IGF-1)等基因表达水平均下降。结论GnRHa除抑制垂体GnRHR产生受体降调节外，还可抑制下丘脑GnRH的基因表达，使性腺激素水平降低，从而减缓第二性征的成熟程度和速度；同时GnRHa通过促进下丘脑SRIH的基因表达，抑制垂体GH和下肢骺软骨IGF-1的基因表达。这可能是Gn—RHa延缓特发性真性性早熟患儿骨骺闭合的机理之一。     

滋阴泻火中药对青春期大鼠子宫与卵巢ER、IGF-Ⅰ、AR基因及蛋白表达的影响

目的:从基因及蛋白水平研究滋阴泻火中药对青春期大鼠子宫、卵巢雌激素受体(ER)、胰岛素生长因子-Ⅰ(IGF-Ⅰ)、芳香化酶(AR)表达的影响,从雌激素和胰岛素生长因子调控及雌激素合成酶途径探讨滋阴泻火中药防治性早熟的机制。方法:将动物分为对照组和中药组,中药组喂饲滋阴泻火中药每次3ml,每日1次,对照组仅给予等量的0.9%NaCl溶液;末次给药24h后剥离子宫称重,并采用实时荧光定量聚合酶链式反应(real-time fluorescence quantitative PCR,FQ PCR)法观察子宫、卵巢ER、IGF-Ⅰ、AR mR-NA的表达,采用免疫组化法观察子宫、卵巢ER、IGF-Ⅰ、AR蛋白的表达。结果:给予滋阴泻火中药干预后,青春期大鼠子宫重量、子宫/体质量比下降,子宫、卵巢ER、IGF-Ⅰ、AR的mRNA和蛋白的表达量均明显下调。结论:滋阴泻火中药能够显著下调靶器官雌激素受体、胰岛素样生长因子及雌激素合成酶(芳香化酶),这可能是其治疗儿童性早熟的作用机制之一。     

疏肝泻火方对雌性性早熟大鼠血清IGF-1、GH、BGP水平的影响

目的:观察疏肝泻火方对雌性性早熟大鼠身长、胫骨长度和血清胰岛素样生长因子-1(IGF-1)、生长激素(GH)、骨钙素(BGP)水平的影响。方法:取SD雌性大鼠60只,随机分为正常对照组、性早熟空白组、亮丙瑞林组以及疏肝泻火高、中、低剂量组6组,除正常对照组外,其余组大鼠采用兴奋性氨基酸-N-甲基-DL-天冬氨酸(NMA)诱导建立性早熟模型,并分别予药物干预,观察大鼠顶臀径、胫骨长度及黄体出现情况,测定血清IGF-1、GH、BGP水平。结果:与性早熟空白组比较,正常对照组、亮丙瑞林组、疏肝泻火高剂量组大鼠顶臀径较小(P0.05);与性早熟空白组比较,亮丙瑞林组与疏肝泻火高、中剂量组血清GH、IGF-1、BGF的浓度均较低(P0.05),低剂量组血清GH、IGF-1浓度较低(P0.05),但高于亮丙瑞林组(P0.05)。结论:疏肝泻火方改善性早熟患儿成年身高的机制可能与降低血清GH、IGF-1、BGF的浓度有关,且治疗效果与疏肝泻火方剂量相关。     

滋阴泻火方对性早熟大鼠血清类胰岛素样生长因子-Ⅰ水平的影响

目的探讨滋阴泻火方对性早熟模型大鼠血清类胰岛素样生长因子-Ⅰ(IGF-Ⅰ)水平的影响。方法26日龄雌性SD大鼠36只,随机分成6组,即正常对照组、性早熟模型组、亮丙瑞林组及中药大、中、小剂量组。除正常对照组外,其余各组均采用N-甲基-DL-天冬氨酸(NMA)40 mg.kg-1皮下注射建立性早熟模型,其中亮丙瑞林组用亮丙瑞林干预,中药各组用不同剂量的滋阴泻火方干预,观察不同干预措施对性早熟模型大鼠卵巢指数、子宫指数及血清IGF-Ⅰ水平的影响。结果性早熟模型组和中药小剂量组的卵巢指数[分别为(6.37±1.61)×10-4、(5.10±2.59)×10-4]和子宫指数[分别为(8.91±2.32)×10-4、(7.86±1.98)×10-4]均显著高于其他各组(P<0.05);亮丙瑞林组、中药大剂量组和中药中剂量组卵巢指数和子宫指数与正常对照组相比均无统计学差异(P>0.05)。与正常对照组比较,性早熟模型组的血清IGF-Ⅰ水平显著升高[(327.08±46.15)vs(239.40±61.92)m IU.m l-1,P<0.05],而4个干预组均无显著性差异(P>0.05);与性早熟模型组比较,中药大剂量组[(218.83±77.06)m IU.m l-1]和中剂量组[(207.09±53.00)m IU.m l-1]的血清IGF-Ⅰ水平显著降低(P<0.05),中药小剂量组无显著性差异(P>0.05);中药各剂量组与亮丙瑞林组比较无显著性差异(P>0.05)。结论滋阴泻火方大、中剂量能降低性早熟模型大鼠的血清IGF-Ⅰ的水平。     

谷氨酸单钠对大鼠骨发育的影响

观察出生１０ｄ内皮下注射ＭＳＧ的Ｗｉｓｔａｒ大鼠，新生期（１１日龄）、青春期（５２日龄）、成年期（１００日龄）长骨发育情况。结果表明：青春期及成年期上、下肢各长骨骨干长度、胫骨骺板厚度及骺板增殖区厚度，ＭＳＧ组均落后于对照组（Ｐ＜０．０５或０．０１），１１日龄时两组长骨骨干长度虽无显著性差异，但决定长骨生长速度的骺板增殖区厚度已出现了显著性差异。各年龄期ＭＳＧ组骺板结构及骨生长发育过程无明显异常，仅厚度变薄。有别于因维生素类、甲状旁腺素等因素引起的生长迟缓。结合各年龄期ＭＳＧ处理鼠垂体湿重分别为对照组的５５．５６％、４２．３９％和４７．７０％，ＭＳＧ处理鼠身长、体重均比对照鼠低２个标准差以下。认为骨的发育及骺板变化符合ＧＨ分泌减少所致。     

滋阴泻火中药对青春期大鼠子宫与卵巢ER、IGF-I、AR基因及蛋白表达的影响

目曲：从基因及蛋白水平研究滋阴泻火中药对青春期大鼠子宫、卵巢雌激素受体（ER）、胰岛素生长因子-I（IGF-I）、芳香化酶（AR）表达的影响,从雌激素和胰岛素生长因子调控及雌激素合成酶途径探讨滋阴泻火中药防治性早熟的机制。方法：将动物分为对照组和中药组,中药组喂饲滋阴泻火中药每次3ml,每日1次,对照组仅给予等量的0．9％NaCl溶液;末次给药24h后剥离子宫称重,并采用实时荧光定量聚合酶链式反应（real-time fluorescence quantitativePCR,rQPCR）法观察子宫、卵巢ER、IGF-I、ARmR-NA的表达,采用免疫组化法观察子宫、卵巢ER、IGF-I、AR蛋白的表达。结果：给予滋阴泻火中药干预后,青春期大鼠子宫重量、子宫／体质量比下降,子宫、卵巢ER、IGF-I、AR的mRNA和蛋白的表达量均明显下调。结论：滋阴泻火中药能够显著下调靶器官雌激素受体、胰岛素样生长因子及雌激素合成酶（芳香化酶）,这可能是其治疗儿童性早熟的作用机制之一。     

抗早2号方对青春期雌性大鼠骨骼生长的影响

目的:研究抗早2号方对青春期雌性大鼠骨骼生长的影响和作用机制。方法:120只SD大鼠随机分为治疗组、中药对照组、西药组、空白组4组。用药4周,停药2周,每3周采样1次,观察抗早2号方对青春期雌性大鼠胫骨长度、宽度、骨密度及尿吡啶酚(PYD)和血清I型胶原前胶原C端肽(PICP)的影响。结果:停药2周时,治疗组与空白组比较,血清PICP降低(P<0.05),尿中PYD含量减少(P<0.05),股骨密度、腰椎骨密度、胫骨长径、宽径差异均无统计学意义(P>0.05);与中药对照组比较,血清PICP增高(P<0.05),尿中PYD含量无明显差异(P>0.05),股骨密度显著降低(P<0.01),腰椎骨密度降低(P<0.05);与西药组比较,血清PICP、尿中PYD、股骨和腰椎骨密度、胫骨长径、宽径差异均无统计学意义(P>0.05)。结论:抗早2号方能抑制青春期雌性大鼠骨骼的快速生长,同时维持骨矿含量的稳定。     

中药调整性早熟儿童青春发育进程的机制研究

目的:从神经内分泌调节及基因表达的角度,探讨调整性早熟儿童青春发育进程的滋阴泻火中药和益肾填精中药有效调节下丘脑垂体促性腺机能、促生长机能的作用机制。方法:青春期大鼠分别予以喂饲滋阴泻火中药或益肾填精中药。采用神经生物学实验方法(下丘脑推挽灌流、组织匀浆、脑片孵育及免疫组化法)检测下丘脑促性腺区促性腺激素释放激素(gonadotropinreleasinghormone,GnRH)的含量、脉冲释放频率与幅度,以及中枢氨基酸递质、神经肽Y及β内啡肽释放的变化;采用定量逆转录聚合酶链反应检测下丘脑GnRH、生长激素释放激素(growthhormonereleasinghormone,GHRH)、生长抑素(somatostatin,SS)、腺垂体卵泡刺激素(follicularstimulatinghormone,FSH)、黄体生成素(luteinizinghormone,LH)、生长激素(growthhormone,GH)、长骨干骺端胰岛素样生长因子Ⅰ(insulinlikegrowthfactorⅠ,IGFⅠ)基因及蛋白的表达水平。结果:滋阴泻火中药可明显抑制下丘脑促性腺区兴奋性氨基酸递质的释放,促进抑制性氨基酸递质、神经肽Y和β内啡肽的释放,使下丘脑GnRH神经元的功能活动降低,下调GnRH、FSH及LH基因的表达水平,上调下丘脑SS基因的表达,并下调垂体GH及长骨干骺端IGFⅠ基因的表达,从而明显抑制下丘脑垂体的促性腺机能及促生长机能。益肾填精中药则可明显抑制下丘脑促性腺区神经肽Y的释放,使下丘脑GnRH神经元的功能活跃,上调GnRH、FSH及LH基因的表达,下调下丘脑SS基因的表达,上调垂体GH及长骨干骺端IGFⅠ基因的表达,从而明显促进下丘脑垂体的促性腺机能及促生长机能。结论:滋阴泻火中药和益肾填精中药可通过调整机体的神经内分泌调节机制,调整下丘脑GnRH、SS,腺垂体FSH、LH、GH及长骨干骺端IGFⅠ基因的转录,有效调节下丘脑垂体的促性腺机能及促生长机能。这可能是滋阴泻火中药和益肾填精中药可有效调整性早熟患儿青春发育进程及改善骨骼发育的主要作用机制。     

滋阴泻火方对性早熟女童骨代谢影响的临床研究

[目的]探讨滋阴泻火方对中枢性性早熟女童骨代谢的影响.[方法]选取2018年10月—2020年1月就诊于南京中医药大学附属医院生长发育专科门诊,经GnRHa激发试验诊断为特发性中枢性性早熟(ICPP)女童80例,其中中药组45例,西药组35例,中药组予滋阴泻火方为基础治疗,西药组予促性腺激素释放激素类似物(GnRHa)...     

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.