三氧化二砷与顺铂联用对肝癌HepG2细胞的作用

目的 探讨三氧化二砷(As2O3)与顺铂(CDDP)联合对肝癌细胞HepG2的作用.方法 应用MTT法和细胞免疫组化酶法检测人肝癌HepG2细胞Bax,Bcl-2,Fas蛋白的表达情况.结果 As2O3与CDDP联合应用在抑制肝癌细胞的生长繁殖、诱导肝癌细胞凋亡,增强Bax,Fas的表达,下调Bcl-2的表达方面均较各自单用的作用明显增强.结论 As2O3与CDDP联合应用具有明显的协同抗肝癌作用,其主要机制可能为As2O3联合化疗能诱导肝癌细胞凋亡,与增强Bax,Fas的表达,下调Bcl-2的表达有关.     

光动力作用对膀胱癌细胞线粒体相关调控蛋白Bcl-2/Bax表达的影响

目的探讨激光活化BPD-MA光动力诱导肿瘤细胞凋亡的可能机制.方法应用免疫组织化学染色检测激光活化BPD-MA光动力作用后人膀胱癌细胞BIU-87线粒体相关调控蛋白Bcl-2和Bax蛋白表达水平.结果激光活化BPD-MA光动力作用后膀胱癌细胞线粒体相关调控蛋白Bcl-2表达显著低于对照组(P<0.05),而Bax蛋白表达水平无明显改变(P>0.05),但Bax/Bcl-2 比值较对照组明显上升(P<0.05).结论激光活化BPD-MA光动力作用后人膀胱癌细胞BIU-87线粒体相关调控蛋白Bcl-2表达水平的下降、Bax/Bcl-2比值的上升.激光活化BPD-MA光动力作用可能通过调控线粒体相关调控蛋白Bcl-2和Bax蛋白表达水平诱导人膀胱癌细胞株BIU-87发生凋亡.     

山楂叶总黄酮对H2O2诱导PC12细胞凋亡的保护作用及机制的研究

目的探讨山楂叶总黄酮(flavone mixture of crataegus leaves,FMCL)对H2O2诱导的PC12细胞凋亡的保护作用及其分子机制.方法用H2O2刺激PC12细胞使其发生凋亡,用流式细胞仪检测PC12细胞凋亡率,用Western blot 法检测Bcl-2、Bax、Caspase-3P20蛋白及用RT-PCR法检测Bcl-2、Bax mRNA表达量的变化.结果与模型组比较FMCL 100μg/ml剂量组PC12细胞凋亡率下降,Bcl-2蛋白及mRNA表达量明显升高,而Bax、Caspase-3P20蛋白及Bax mRNA表达量明显降低(P<0.05或P<0.01),且在一定范围内存在剂量依赖关系.结论山楂叶总黄酮可抑制H2O2诱导的PC12细胞凋亡,其机制可能与调节凋亡相关基因及蛋白的表达从而抑制Caspase-3活化有关.     

三氧化二砷联合氟尿嘧啶对人肝癌SMMC-7721细胞Bcl-2和Bax表达的影响

目的探讨三氧化二砷联合氟尿嘧啶（5-FU）对肝癌细胞凋亡相关因子Bcl—2和Bax表达的影响。方法体外培养人肝癌SMMC-7721细胞株,将细胞分为四组,即AS2O3组、5-FU组、As2O3＋5-Fu组及对照组,采用免疫细胞化学法检测各组SMMC-7721细胞Bcl-2、Bax蛋白的表达。结果As2O3组、5-Fu组、As2O3＋5-FU组细胞Bcl-2蛋白表达较对照组明显降低,Bax蛋白表达明显升高（P〈0．01）;联合用药组细胞Bcl-2蛋白表达较单用药组明显降低,Bax蛋白表达明显升高（P〈0．01）。结论As2O3联合5-FU可下调Bcl-2蛋白的表达,上调Bax蛋白的表达,从而诱导肝癌细胞凋亡。     

中华眼镜蛇毒组分对MCF-7乳腺癌细胞凋亡的诱导作用及对Bcl-2、Bax蛋白表达的影响

目的探讨中华眼镜蛇毒组分(naja naja actra venom component,NNAVC)对MCF-7乳腺癌细胞凋亡的诱导作用及对Bcl-2、Bax蛋白表达的影响。方法采用MTT法检测不同浓度(0.312 5、0.625、1.252、.5、5、10μg/mL)、不同作用时间(244、8、72 h)下NNAVC对MCF-7细胞的增殖抑制作用;倒置显微镜观察NNAVC对MCF-7细胞形态学影响;流式细胞仪检测细胞凋亡情况;Western blot法检测Bcl-2及Bax蛋白的表达。结果 MTT结果显示,NNAVC对MCF-7细胞具有明显的增殖抑制作用,且呈时间及剂量依赖性。倒置显微镜观察NNAVC作用MCF-7细胞后,细胞出现形态不规则、部分细胞变圆甚至脱落、细胞碎片等改变。流式细胞仪检测2.5μg/mL和5μg/mL的NNAVC均明显诱导MCF-7细胞凋亡,凋亡率分别达到(35.08±4.24)%和(44.41±3.56)%。Western blot法结果显示Bcl-2蛋白表达减少,而Bax蛋白表达增多。结论 NNAVC对MCF-7细胞具有明显的增殖抑制作用,且可以诱导其凋亡,其诱导凋亡机制可能与下调Bcl-2和上调Bax蛋白表达有关。     

五倍子单宁酸体外诱导人膀胱癌EJ细胞凋亡及机制研究

目的观察五倍子单宁酸(TA)在体外培养条件下对人膀胱癌EJ细胞增殖、凋亡的影响及其作用机制。方法将EJ细胞分为阴性对照组(TA浓度为0μmol/L)和不同梯度浓度TA实验组,阴性对照组加入不含TA药物的培养基,其他各组加入含有梯度浓度TA的培养基;用台盼蓝染色检测细胞损伤率,CCK-8检测细胞增殖率,吖啶橙/溴化乙锭(AO/EB)双荧光染色以及流式分析检测细胞凋亡率,Western Blot(WB)检测细胞内半胱氨酸天冬氨酸蛋白酶-3(Caspase-3),BCL2-AssociatedX蛋白(Bax),B淋巴细胞瘤-2(Bcl-2)蛋白表达。结果与阴性对照组比较,实验组TA可降低EJ细胞活力,抑制细胞增殖,诱导细胞凋亡,促进细胞内Caspase3表达、降低Bcl-2蛋白表达、增加Bax/Bcl-2比值差异均具有统计学意义(P0.05)。结论TA可能通过调节细胞内Caspase3、Bax、Bcl-2蛋白表达诱导EJ细胞凋亡,抑制增殖,提示TA可能是一种潜在的抗膀胱癌药物。     

三氧化二砷诱导肿瘤细胞凋亡与Caspase3及Bcl-2蛋白家族的关系

目的：探讨三氧化二砷（As2O3)诱导肿瘤细胞凋亡的机制。方法：以Namalwa,SGC7901和Bcap37细胞为体外模型，通过流式细胞仪检测亚G1期（sub－G1）细胞含量和细胞膜磷脂酰丝氨酸（phosphatidylserine,PS)外翻量鉴定细胞凋亡；流式细胞仪检测Bcl-2和Bax蛋白表达及细胞线粒体跨膜电位（△φm）；比色法8测定半胱氨酸酶3（Caspase3）活性。结果：As2O3诱导肿瘤细胞凋亡与Bax和Bcl-2蛋白表达及Caspase3活性有密切关系。结论：As2O3可能通过调节Bax和Bcl－2蛋白的表达引起△φm下降从而激活Caspase3最终导致肿瘤细胞凋亡。     

三氧化二砷对宫颈癌Hela细胞体外生长的影响

目的 研究1.0、2.0和3.0 μmol/L的三氧化二砷(As2O3)对人宫颈癌Hela细胞体外生长的影响,并初步探讨其机制.方法 应用1.0、2.0和3.0 μmol/L As2O3作用于人宫颈癌Hela细胞系,采用MTT比色法检测细胞生长情况;运用流式细胞术检测细胞凋亡率;应用免疫组化SABC法检测细胞中Bcl-2和Bax蛋白的表达.结果 As2O3能抑制宫颈癌Hela细胞的体外生长,诱导宫颈癌Hela细胞凋亡,并呈浓度和时间依赖性(P＜0.05).As2O3能引起细胞内Bcl-2蛋白表达下调和Bax蛋白表达上调(P＜0.05).结论 As2O3对宫颈癌Hela细胞体外生长具有明显抑制作用,并诱导宫颈癌Hela细胞凋亡,可能与其引起细胞内Bcl-2蛋白表达下调和Bax蛋白表达上调有关.     

祁连圆柏提取物诱导人肝癌细胞Bel-7402凋亡的作用及筛选研究

目的探讨祁连圆柏提取物体外对人肝癌细胞Bel-7402体外增殖抑制、凋亡作用机制并进行有效组分筛选研究。方法采用硅胶柱层析法提取祁连圆柏有效抗瘤组分,分光光度法测定含量。用克隆形成实验、台盼蓝拒染法及MTT显色法检测祁连圆柏提取物对Bel-7402细胞增殖的影响,流式细胞术检测其诱导细胞凋亡的细胞周期阻滞状况及凋亡率,免疫组化观察对Bcl-2与Bax蛋白的表达水平,光镜、电镜观察凋亡细胞的组织形态和超微结构变化。结果祁连圆柏提取物对Bel-7402细胞增殖有明显的抑制作用,其中C液-组分-3细胞的半数抑制浓度为2.47μg/ml。流式细胞检测结果 G0/G1期细胞增多,G2/M期细胞减少,其中10～50μg/ml的祁连圆柏C液-组分-3在G1期细胞前出现典型"凋亡峰";免疫组化SP法检测实验组细胞BCL-2基因蛋白表达降低,Bax基因蛋白表达增加。光镜与电镜下均见凋亡细胞明显增多。结论祁连圆柏C液-组分-3具有显著的抑制Bel-7402细胞增殖以及诱导该细胞系凋亡的作用,其诱导Bel-7402细胞凋亡的作用与其上调Bax、下调Bcl-2蛋白的表达,激活Bax凋亡基因蛋白及G0/G1期阻滞有关,是其重要的抗肿瘤作用机制。     

胸腺肽α1诱导膀胱癌细胞株BIU87凋亡的实验研究

目的研究胸腺肽α1诱导膀胱癌细胞株BIU87凋亡的作用及其机制。方法用50~400μg/ml胸腺肽α1作用BIU87细胞24~72 h后,MTT比色法检测细胞生长活性,单细胞凝胶电泳法和流式细胞仪分别检测BIU87细胞DNA损伤和凋亡作用。结果胸腺肽α1能显著抑制膀胱癌细胞株BIU87细胞的体外生长(P<0.01),呈时间与剂量依赖性;流式细胞仪检测显示不同浓度凋亡率分别为(4.69±0.60)%、(21.15±3.10)%、(33.15±2.40)%、(40.87±1.50)%、(48.47±1.02)%;单细胞凝胶电泳检测显示不同浓度DNA损伤比率分别为5%、20%、41%、62%、85%。结论胸腺肽α1通过抑制BIU87细胞增殖、诱导细胞凋亡等机制,显著抑制膀胱癌细胞生长。     

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.