少弱精子症患者睾丸组织中NOSTRIN的表达及其与eNOS的相关性研究

目的 通过测定少弱精子症患者睾丸组织中内皮型一氧化氮合酶运输介导物(endothelial nitric oxide syn-thase traffic inducer,NOSTRIN)和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)的表达,探讨其与少弱精子症发病的相关性.方法 采用逆转录一聚合酶链反应(RT-PCR)检测11例严重少弱精子症患者(病例组)和7例正常男性(正常组)睾丸组织中NOSTRIN mRNA的表达;分光光度法测定睾丸组织中eNOS的活性;应用硝酸还原酶法测定睾丸组织上清液中NO代谢产物NO2-/NO3-水平.结果 病例组患者睾丸组织中NOSTRIN mRNA呈弱表达(0.312±0.076),明显低于正常组(0.793±0.082,P<0.01);病例组患者睾丸组织eNOS活性[(33.73±3.58)U/mg]与正常组[(17.69±3.84)U/mg]比较显著增强(P<0.01);病例组患者睾丸中NO2-/NO3-水平为(48.56±8.49)μmol/L,较正常组(25.37±9.61)μmol/L显著升高(P<0.01)f病例组睾丸组织中NOSTRIN表达水平与eNOS活性呈负相关(r=-0.57,P<0.01),与NO代谢产物NO2-/NO3-同样呈负相关(r=-0.61,P<0.01).结论 少弱精子症患者睾丸组织中NOSTRIN表达水平降低,eNOS活性增强,NO2-/NO3-水平升高,可能与少弱精子症的发生有着相关性.     

复方生精汤治疗少弱精子症男性不育临床研究

目的:探讨复方生精汤治疗少弱精子症男性不育的临床疗效。方法:选择2014年1月至2017年12月收治的男性不育症患者60例,随机分为观察组与对照组各30例。观察组采用复方生精汤治疗,对照组采用六味地黄丸治疗。比较两组患者临床疗效、精子存活率、精子密度、精子活力、性激素水平。结果:观察组治愈及总有效人数多于对照组,差异有统计学意义(P0.05);治疗后两组精子密度、精子存活率、a级活力以及a级+b级活力均显著高于治疗前,差异有统计学意义(P0.05),且观察组精子存活率、a级活力以及a级+b级活力显著高于对照组,差异有统计学意义(P0.05);治疗后两组患者性激素指标较治疗前均有改善(P0.05),但组间比较差异无统计学意义(P 0.05)。结论:复方生精汤可通过提高精子质量、改善激素水平治疗少弱精子症男性不育。     

麒麟丸联合左氧氟沙星治疗伴有慢性前列腺炎的少弱精子症的临床疗效观察

目的:观察麒麟丸联合左氧氟沙星治疗伴有慢性前列腺炎的少弱精子症临床疗效及安全性。方法:将伴有慢性前列腺炎的少弱精子症患者90例随机分为治疗组与对照组,每组各45例。对照组口服左氧氟沙星,服用3周;治疗组在上述治疗基础上,同时使用口服麒麟丸,4周为1疗程。每组在治疗前和治疗1疗程后做精液分析,以精子浓度和a级精子百分率、(a+b)级精子百分率、精子活动率以及观察患者治疗后的治愈率、总有效率和前列腺症状指数表(NIH-CPSI)评分为主要疗效指标,评价治疗效果。结果:两组患者的精液常规各项参数及治愈率和总有效率比较,差异均有统计学意义(P<0.01);两组患者NIH-CPSI评分均有改善,但改善幅度比较差异无统计学意义(P>0.05)。结论:治疗伴有慢性前列腺炎的少弱精子症,麒麟丸联合左氧氟沙星可明显提高精液质量,且比单独应用左氧氟沙星具有更好的疗效,未见明显不良反应。     

少弱精子症和弱精子症患者精液质量下降与锌的关系

采用原子吸收光谱法对67例患弱精子症和少弱精子症的男性不育症患者精子和精浆中锌元素进行了测定,结果发现弱精子症患者和少弱精子症患者精子中锌的含量明显低于正常具有生育能力的对照组,统计学上有极显著性差异(P<0.01)。而精浆中锌的含量三组间差别无统计学意义(P>0.05)。研究提示弱精子症患者和少弱精子症患者的精子运动能力降低以及精子的数量减少可能与精子中锌元素的含量下降有关。对这类患者酌情补锌具有一定的治疗意义。     

弱精子症患者精子中DKKL1的表达

目的探讨精子顶体相关基因（DKKL1）在弱精子症患者精子中的表达特征及临床意义。方法收集正常男性和弱精子症 患者精液,各取10 μL精液用全自动计算机辅助精液分析系统（CASA）对精液进行常规分析。为了避免白细胞和生精细胞对结 果的影响,剩余精液采用4个梯度的Percoll法离心分离和纯化精子。Trizol法提取精子RNA,实时荧光定量PCR从基因水平检 测DKKL1基因在正常人和弱精子症患者精子中的表达差异;最后采用Western blot方法从蛋白水平检测DKKL1蛋白在正常人 和弱精子症患者精子中的表达差异。结果精液常规分析结果显示,弱精子症和正常人两组之间的年龄、精液体积、精子浓度和 正常精子形态比例均没有统计学差异（P>0.05）。与正常对照组相比,弱精子症组PR 和PR+NP 精子的比例显著降低（P< 0.01）。实时荧光定量PCR结果显示,与正常对照组相比,DKKL1 mRNA在弱精子症患者精子中的表达量降低11.1倍。此外, Western blot结果显示,弱精子症组DKKL1蛋白的表达量与正常对照组相比,降低了2.4倍,差异具有统计学意义（P<0.01）,该 结果与实时荧光定量PCR结果一致。结论DKKL1在弱精子症患者精子中的表达水平明显下降,表明其与精子运动密切相关, 是导致弱精子症发生的原因之一。     

多潘立酮对胃黏膜的保护作用探讨

目的探讨多潘立酮对胃黏膜的保护作用。方法将大鼠随机分为对照组和实验组。对照组予生理盐水1.5ml灌胃,实验组予多潘立酮1mg/kg灌胃,3次/d,连续3d。放射免疫法测定两组胃黏膜前列腺素E2(PGE2)水平;免疫组织化学方法检测胃黏膜细胞中环氧合酶(COX)-1和COX-2蛋白表达水平并以平均吸光度值表示其强度;逆转录聚合酶链反应(RT-PCR)检测胃黏膜COX-1和COX-2基因表达变化。结果实验组和对照组胃黏膜PGE2含量分别为(81.93±35.58)pg/μl、(237.14±79.62)pg/μl,两组比较差异具有统计学意义(t=46.83,P<0.01);实验组和对照组胃黏膜COX-1蛋白表达水平(吸光度值)分别为0.209±0.034、0.232±0.036,两组比较差异具有统计学意义(t=10.56,P<0.01);实验组与对照组胃黏膜COX-1 mRNA表达水平分别为0.650±0.056、1.540±0.081,两组比较差异具有统计学意义(t=7.35,P<0.05)。实验组和对照组大鼠的胃黏膜细胞内均无COX-2蛋白以及COX-2 mRNA表达。结论多潘立酮对胃黏膜具有保护作用,可能与其促进胃黏膜PGE2合成,增加胃黏膜COX-1基因和蛋白表达有关。     

特发性弱精子症患者精子DNA氧化损伤

目的:探讨特发性弱精子症患者精浆的氧化应激及其精子DNA的氧化损伤。方法:选择2010年12月至2011年3月就诊于中南大学湘雅医院生殖中心门诊的不孕不育夫妇。以28名特发性弱精子症患者为实验组,选取24名精液分析正常且有生育史的男性为对照组,收集新鲜精液。应用鲁米诺化学发光法检测原始精浆活性氧(reactive oxygen species,ROS)水平;酶联免疫分析法检测精子DNA氧化产物8-羟基脱氧鸟苷(8-hydroxy-2'-deoxyguanosine,8-OHdG)浓度。结果:1) 实验组精浆ROS水平增高,与对照组比较差异有统计学意义(P<0.01);两组前向运动精子活动率与精浆ROS水平呈负相关(r=-0.72,P<0.01)。2) 实验组精子8-OHdG浓度高于对照组,差异有统计学意义(P<0.01);两组前向运动精子活动率与精子8-OHdG浓度也呈负相关(r=-0.73,P<0.01)。3) 两组患者精浆ROS水平与精子8-OHdG浓度呈正相关(r=0.77,P<0.01)。结论:特发性弱精子症患者的精子具有DNA氧化损伤,可能与氧化应激有关。过量活性氧产生可能是特发性弱精子症患者精子活力低下的原因之一。     

麒麟丸治疗男性不育症的临床疗效观察

目的观察麒麟丸治疗由少弱精子症引起男性不育症的临床疗效。方法选择2015年8月至2016年8月收治的不孕不育症患者210例,其中由于男性少弱精子症原因引起者共83例(39.5%),所有患者根据就诊顺序以随机数字表法分为对照组(37例)与治疗组(46例)。对照组患者给予左卡尼汀口服液治疗,治疗组患者采用麒麟丸联合左卡尼汀口服液治疗。两组患者均治疗3个月为1个疗程,治疗1个疗程后观察两组患者的精子浓度、a级精子百分率、(a+b)级精子百分率以及精子活动率情况。同时统计患者配偶的妊娠率。结果 83例患者中对照组1例,治疗组3例未及时复诊计入脱落病例。两组患者治疗前精子浓度、a级精子百分率、(a+b)级精子百分率以及精子活动率情况比较,差异无统计学意义(P0.05);两组治疗3个月后精子浓度、a级精子百分率、(a+b)级精子百分率以及精子活动率情况与治疗前比较,差异有统计学意义(P0.05),且治疗组精液各指标改善情况以及妊娠率均明显优于对照组(P0.05,P0.01)。结论麒麟丸联合左卡尼汀联合治疗由少弱精子症引起男性不育症疗效显著,可能与麒麟丸促进精子的形成有密切相关性,而且可显著提高患者的精液质量,提示中西医结合治疗少弱精子不育症患者具有明显优势,值得推广。     

麒麟丸联合左氧氟沙星治疗伴有慢性前列腺炎的少弱精子症的临床疗效观察

目的:观察麒麟丸联合左氧氟沙星治疗伴有慢性前列腺炎的少弱精子症临床疗效及安全性。方法:将伴有慢性前列腺炎的少弱精子症患者90例随机分为治疗组与对照组,每组各45例。对照组口服左氧氟沙星,服用3周;治疗组在上述治疗基础上,同时使用口服麒麟丸,4周为1疗程。每组在治疗前和治疗1疗程后做精液分析,以精子浓度和a级精子百分率、(a+b)级精子百分率、精子活动率以及观察患者治疗后的治愈率、总有效率和前列腺症状指数表(NIH-CPSI)评分为主要疗效指标,评价治疗效果。结果:两组患者的精液常规各项参数及治愈率和总有效率比较,差异均有统计学意义(P0.01);两组患者NIH-CPSI评分均有改善,但改善幅度比较差异无统计学意义(P0.05)。结论:治疗伴有慢性前列腺炎的少弱精子症,麒麟丸联合左氧氟沙星可明显提高精液质量,且比单独应用左氧氟沙星具有更好的疗效,未见明显不良反应。     

下肢静脉曲张患者外周血Nelin水平测定及意义

目的 探讨下肢静脉曲张患者与正常人外周血中Nelin表达水平的差异及其意义。方法 选取下肢静脉曲张患者57例为对照组,健康查体者60例为正常组,利用RT-PCR技术检测外周血单个核细胞中Nelin mRNA的表达水平,利用生物素双抗体夹心ELISA技术测定外周血血清中Nelin蛋白的表达水平。结果 RT-PCR结果显示,Nelin在对照组与正常组的相对表达量分别是 0.226±0.070、0.718±0.114,两组差异有统计学意义(t=9.028,P<0.01);ELISA结果显示,96例血清中均可检测到Nelin蛋白的表达,对照组和正常组的表达量分别是(2.61±1.73)ng/mL、(3.64±1.65)ng/mL,两组差异有统计学意义(t=2.92,P<0.01)。结论 与正常人相比,下肢静脉曲张患者Nelin基因在转录和翻译水平均存在明显的表达下调,提示Nelin基因的表达可能与下肢静脉曲张发生发展有一定的关系。     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.