脂多糖诱发肝损伤机理的初步研究

目的:探讨LPS肝损伤的信号转导途径及LPS诱导肝细胞凋亡的规律。方法:Balb/c小鼠腹腔注射LPS建立内毒素血症模型,在不同时间点取其肝脏,HE染色观察病理改变,RT-PCR法检测肝组织TLR4的表达。TUNEL法检测肝细胞凋亡,免疫组化法检测bax、bcl-2、Fas、Fasl表达。结果:LPS注入腹腔后24h小鼠肝脏开始出现明显病理改变;3～6h肝细胞凋亡达高峰,12h后明显减少;肝脏TLR4在LPS注射3h后显著下调,6～12h明显受抑制,24h后则基本恢复;LPS刺激后3～6h肝脏bax、Fas表达达高峰,而bcl-2、Fasl表达在6～12h达到高峰,随后都逐渐下降。结论:LPS刺激肝脏后,肝损伤呈时间依从性,凋亡是其细胞死亡方式之一。LPS刺激后,肝脏TLR4表达呈时间依从性,TLR4表达下调可能是对LPS产生的耐受现象,是机体抗损伤的一种保护性的下调。LPS所致肝细胞凋亡是多因素作用的结果。     

LPS诱导鼠肝脏和腹腔巨噬细胞TLR4表达和肝损伤的动态分析

目的观察LPS刺激后不同时间点肝脏和腹腔巨噬细胞的TLR4表达,以及分析TLR4动态变化规律.方法以BALB/c小鼠为模型,腹腔注射LPS后不同时间取肝脏和腹腔巨噬细胞抽提RNA,逆转录-聚合酶链式反应半定量分析TLR4mRNA各时相的表达.体外培养的腹腔巨噬细胞不同剂量LPS刺激后观察TLR4mRNA的表达规律.结果注射LPS 3 h后肝脏和腹腔巨噬细胞TLR4mRNA开始明显下降,第6、12 h基本检测不到,24 h后逐渐恢复,30 h小鼠死亡时基本恢复到正常.不同浓度LPS均下调腹腔巨噬细胞TLR4mRNA的表达,剂量之间差异无显著性.血中ALT在3h开始升高,6h达高峰,肝细胞病理变化出现在12 h,随时间的延长而加重.结论LPS可直接损伤肝细胞,这种损伤与TLR4无关.LPS可能通过TLR4介导肝组织的病理改变.     

LPS信号转导分子TLR4表达与小鼠肝损伤的关系

目的：探讨脂多糖(1ipopolysaccharide，LPS)信号转导分子，TLR4(Toil-like receptor 4,TLR4)与肝脏损伤的关系。方法：小鼠腹腔注射LPS后不同时间点取肝脏组织，HE染色观察病理改变，RT-PCR法检测肝组织，TLR4 mRNA的表达。结果：腹腔注射LPS后24h小鼠肝脏出现明显炎性浸润、坏死，肝组织TLR4 mRNA的表达在注射LPS后6，12h明显抑制，24h则基本恢复。结论：LPS致肝损伤呈时间依从性；LPS信号转导分子TLR4在介导LPS肝损伤中起重要作用。。     

LPS信号转导分子TLR4表达与小鼠肝损伤的关系

目的 :探讨脂多糖 (lipopolysaccharide ,LPS)信号转导分子TLR4(Toll likereceptor 4 ,TLR4)与肝脏损伤的关系。方法 :小鼠腹腔注射LPS后不同时间点取肝脏组织 ,HE染色观察病理改变 ,RT PCR法检测肝组织TLR4mRNA的表达。结果 :腹腔注射LPS后 2 4h小鼠肝脏出现明显炎性浸润、坏死 ,肝组织TLR4mRNA的表达在注射LPS后 6 ,1 2h明显抑制 ,2 4h则基本恢复。结论 :LPS致肝损伤呈时间依从性 ;LPS信号转导分子TLR4在介导LPS肝损伤中起重要作用     

L-精氨酸诱导大鼠暴发性胰腺炎肝损伤的实验研究

目的：建立L-精氨酸诱导大鼠暴发性胰腺炎（FAP）肝损伤模型，并探讨FAP肝损伤机制。方法：L-精氨酸分次间隔2h分别于腹腔和皮下注射。末次注射后观察120h内大鼠死亡率以及6、12、24、48、72和120h血清淀粉酶、谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）以及门静脉血中肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）含量变化；同时观察各时点胰腺、肝脏病理改变及肝细胞凋亡变化。结果：注射后大鼠120h内死亡率为53．33％；AST、ALT、TBIL、IL-6、TNF-α于6h升高，其中AST、TBIL、IL-6于24h达高峰，ALT、TNF-α于48h达高峰；胰腺、肝脏病理改变以及肝细胞凋亡均于6h开始变化，48h达高峰。结论：大剂量L-精氨酸可诱导大鼠FAP肝损伤模型，TNF-α、IL-6、细胞凋亡可能参与其损伤机制。     

缺血预处理对肝缺血再灌注时细胞凋亡及调控基因表达的影响

目的探讨缺血预处理对肝脏细胞凋亡的影响以及与调控基因bcl-2／bax蛋白表达的关系。方法将家兔分为假手术（SO）组、缺血再灌注（IK）组，缺血预处理（IP）组。后2组中再分为4个亚组（IR3h、12h、24h和48h组。IP3h、12h、24h和48h组）。缺血期均为60min。缺血预处理为缺血前采用5min缺血及5min再灌注的方法。分别于再灌注3、12、24和48h后处死动物采取肝脏标本，SO组于手术后24h采取肝标本。检测细胞凋亡及bcl-2／bax蛋白表达水平。结果IR组和IP组与SO组比较，细胞凋亡指数（AI）增加差异有显著性（P〈0.01）；IP组中IP3h、12h和24h较同时间段IR组细胞凋亡指数降低差异有显著性（P〈0.05）；bcl-2蛋白表达：IP组与IR组及SO组比较，增高差异有显著性（P〈0.05）；bax蛋白表达：IR组和IP组与SO组比较增加差异有显著性（P〈0.05）。结论缺血预处理可能通过激活bcl-2蛋白的表达，改变bcl-2／bax表达比例从而抑制肝细胞凋亡。     

白花前胡及前胡甲素对心肌缺血再灌注大鼠IL-6水平及Fas、bax、bcl-2蛋白表达的影响

目的：研究中药白花前胡及其有效成分对缺血再灌注心肌IL-6水平及凋亡相关蛋白表达的影响。方法：开胸麻醉大鼠左冠状动脉前降枝经30min缺血及120min再灌注；放射免疫法测定血清IL-6水平；免疫组化法和计算机图像分析系统检测心肌Fas,bax有bcl-2蛋白表达；苏木精-依红染色法于光镜下观察嗜中性白细胞浸润。结果；白花前胡粗提物及前胡甲素（Pd-Ia）在降压和不影响心率的情况下，减少IL-6水平及Fas，bax,bcl-2蛋白的表达，增加bcl-2/bax的比率。IL-6水平与Fas,bax,bcl-2蛋白表达之间呈密切的线性正相关，而嗜中性白细胞只有轻微浸润。结论：白花前胡及Pd-Ia在防治缺血后心肌细胞死亡上显现新靶位，原因可能与心肌缺血再灌注期间机体自身调控即早基因IL-6,Fas,bax,bcl-2的表达有关。     

暴发性肝衰竭中Fas及caspase-3与肝细胞凋亡的关系

目的研究在暴发性肝衰竭（fulminant hepatic failure，FHF）中Fas及caspase-3表达与肝细胞凋亡的关系。方法联合应用脂多糖（LPS）和D-氨基半乳糖（D—GalN）制备FHF小鼠模型；采用免疫组化方法检测肝组织Fas表达，原位杂交方法检测肝组织caspase-3表达，TUNEL法检测肝组织肝细胞凋亡；在用药后2、4、8和12h动态观察Fas、caspase-3表达及肝细胞凋亡变化。结果在FHF模型小鼠中，用药后8h可出现典型的肝细胞凋亡表现，12h仍存在肝细胞凋亡。用药后2h开始Fas有少量表达，至8h和12h表达均很多，二者比较差异无显著，与2h组比较P〈0．01，与4h组比较P〈0．05。用药后2hcaspase-3少量表达，8h达最高峰，12h较8h减少，8h与2h比较，P〈0．01，与4h和12h比较P〈0．05。Fas和caspase-3表达相一致。结论在FHF中，肝细胞凋亡的发生与Fas及caspase-3的表达增加有关，且Fas介导肝细胞凋亡的过程与caspase-3的激活有关。     

氧化苦参碱对苯巴比妥钠诱导的肝细胞凋亡小鼠肝细胞bcl-2、bax基因表达的影响

目的：探讨氧化苦参碱（OM）阻断苯巴比妥钠（PB）诱导的肝细胞凋亡的机制。方法：105只雄性昆明种小鼠随机分为5组。空白对照组：腹腔注射生理盐水，连续7d，停药12h后处死。PB组：5g/L PB75 mg/kg,1次／d，腹腔注射，连续7d，停药后12h处死。PB撤除组：5g/L PB 75mg/kg,1次／d，腹腔注射，连续6d，停药后36h处死。OM治疗I组：首先腹腔注射5g/L PB 75mg/kg,1次／d；第6d后腹腔注射OM 150mg/kg,3次／d，至停PB后36h，将动物处死。OM治疗Ⅱ组：首先腹腔注射5g/L PB 75mg/kg,1次／d，第6d后腹腔注射OM 75mg/kg,3次／d，至停PB后36h，将所有动物脱颈处死，取出肝脏，体积分数为4％多聚甲醛固定，石蜡包埋。用Tunel法检测肝细胞凋亡，用免疫组化和原位杂交技术检测bcl-2、bax基因的表达。结果：仅PB撤除组Tunel法检测到细胞凋亡。Bcl-2蛋白在OM治疗I、Ⅱ组的表达较PB撤除组显著增强（P＜0．05），在OM治疗I组和Ⅱ组之间差异无统计学意义。Bax蛋白在PB撤除组表达较其他各组显著增强（P＜0．05），而在其他各组之间差异无统计学意义（P＞0．05）。bcl-2、bax基因表达的mRNA阳性信号强度与Bcl-2、Bax蛋白免疫组化反应的强度一致。结论：氧化苦参碱可能通过上调bcl-2基因的表达来阻断肝细胞凋亡。     

对大鼠肝脏缺氧性损伤中凋亡机制的研究

目的：研究凋亡机制在大鼠肝脏缺氧性损伤过程中的作用。方法：应用缺氧法制造大鼠肝脏缺氧模型。对肝组织进行原位缺口末端DNA标记技术检测肝细胞的凋亡情况。用原位杂交及RT－PCR方法检测凋亡相关基因bcl-2和bax的表达。结果：大鼠在连续缺氧（10％O2，8h/d)30天后，缺氧组及对照组肝细胞均有凋亡，但缺氧组凋亡细胞明显减少。bcl-2在缺氧组中表达强度明显高于对照组，而bax在缺氧组中的表达低于对照组。结论：在缺氧因素作用下，bcl-2和bac 2种基因表达比例发生变化，使肝脏组织内肝细胞凋亡减少。     

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.