乳香诱导急性非淋巴细胞白血病细胞凋亡中对Bcl-2基因调节

研究乳香诱导急性非淋巴细胞白血病细胞凋亡中对Bcl-2基因蛋白的影响。采用DNA片段百分率及免疫组化法检测30例急性非淋巴细胞白血病细胞及白血病细胞株HL60细胞经100μg/ml乳香处理前、后Bcl-2基因蛋白表达水平。结果：乳香具有诱导急性非淋巴细胞白血病细胞及HL60细胞凋亡^[1]及下调Bcl-2基因蛋白表达水平。结论：乳香具有诱导急性非淋巴细胞白血病细胞及HL60细胞凋亡作用及下调Bcl-2基因蛋白。Bcl-2基因蛋白表达水平下调可能是乳香提取物诱导急性非淋巴细胞白血病细胞凋亡重要机制之一。     

熊果酸诱导人急性白血病细胞凋亡及其作用机制

目的 探讨熊果酸诱导人急性白血病细胞凋亡的作用机制.方法 将0、5、10、15、20 μmol/L的熊果酸作用于U937细胞6、12 h,观察昔效关系.用20 μmol/L熊果酸作用U937细胞1、3、6、9、12、24 h,观察时效关系.采用AnnexinV/PI染色和流式细胞仪检测细胞凋亡.用Western blot法检测细胞凋亡相关蛋白PARP、Caspase-3、Caspase-9及Bcl-2家族基因Mcl-1、Bcl-2、Bcl-xL、Bax、Bad的表达,用β-actin做内参照.20 μmol/L熊果酸作用于不同类型的急性自血病细胞(U937、Jurkat、HE60)12 h,用流式细胞仪检测细胞凋亡,并用Western blot方法 检测Caspase-3、Caspase-9、PARP的表达.结果 熊果酸作用于U937细胞引起细胞发生凋亡,并呈明显的量效和时效关系.Western blot结果表明,熊果酸引起凋亡蛋白酶Caspase-3、Caspase-9的降解/活化增加,促进凋亡作用底物PARP的降解增加.熊果酸还可引起抗凋亡蛋白Mcl-1的表达降低,但对Bcl-2、Bcl-xL、Bax、Bad的表达均无明显影响.结论 熊果酸可诱导多种急性白血病U937、Jurkat、HL60细胞发生凋亡.Mcl-1的下调可能在熊果酸诱导急性白血病细胞凋亡中起着重要的作用,其作用机制可能为熊果酸引起Mcl-1的下调,随后引起凋亡相关蛋白Caspase-3、Caspase-9的降解/活化及PARP的降解,最终促进细胞凋亡发生.     

青蒿琥酯对不同急性白血病细胞株凋亡抑制的实验研究

[目的]观察青蒿琥酯对不同急性白血病细胞株的增殖抑制作用及凋亡机制探讨。[方法]应用不同浓度青蒿琥酯作用U937、HL60、Jurkat细胞株,MTT法检测抑制率;细胞形态学、DNA琼脂糖凝胶电泳、流式细胞术检测细胞凋亡;Western-blot法检测细胞凋亡过程中Bcl-2、Bax、Caspase 8和ICAD蛋白表达的变化。[结果]青蒿琥酯能够明显抑制U937、HL60、Jurkat细胞增殖;8μg/ml以上浓度青蒿琥酯作用三种细胞24h、48h、72h抑制率均为Jurkat〉HL60〉U937(均P<0.01);其增殖抑制作用与诱导细胞凋亡有关;随药物浓度的增高Bcl-2和ICAD蛋白表达量下降,而Bax蛋白表达上调,Caspase8蛋白出现明显的剪切激活带。[结论]青蒿琥酯既能通过线粒体途径又能通过外源性途径诱导急性白血病细胞凋亡,对Jurkat的抑制最强。     

地西他滨对伊马替尼耐药细胞株KBM5-T315I的增殖抑制及诱导凋亡作用研究

目的探讨地西他滨对伊马替尼耐药细胞株KBM5-T315I的增殖抑制及诱导凋亡作用。方法应用细胞增殖及毒性检测法（新型四唑单钠盐法）观察不同浓度（0—40μmol／L）地西他滨作用于KBM5-T315I细胞24、48及72h后的细胞增殖活力变化。采用AnnexinV-FITC／P[双染流式细胞术观察不同浓度地西他滨对KBM5-T315I处理24h后的细胞凋亡率。JC-1法检测细胞线粒体膜电位的变化,Westernblot检测cleavedPARP的变化。结果地西他滨可显著抑制KBM5-T315I细胞生长,显示出明显的量-效与时-效关系。随着地西他滨药物浓度增加,KBM5-T315I细胞株凋亡率逐渐升高。线粒体膜电位逐渐下降,cleavedPARP表达水平显著上升。结论地西他滨能抑制KBM5-T315I细胞的生长及诱导其凋亡,从而为治疗伊马替尼耐药慢性粒细胞白血病提供实验依据。     

维生素C联合硼替佐米对人T细胞淋巴瘤细胞株Jurkat细胞凋亡的应用

目的 研究维生素C联合硼替佐米对人T细胞淋巴瘤细胞株Jurkat细胞凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)与Bcl2相关X蛋白(Bax)表达调节的影响.方法 采用30 nmol/L的硼替佐米组,100 μg/mL的维生素C组及联合组分别作用于Jurkat细胞24、48、72 h,设立未加药物的对照组,应用倒置显微镜观察细胞的生长方式,免疫组织化学检测Bax及Bcl-2的表达,CCK-8及流式细胞技术检测Jurkat细胞凋亡率和细胞周期.结果 维生素C使Jurkat细胞的生长方式从对照组的局部聚集向近乎均匀分散变化;维C组及联合组在24 h至48 h期间,Bcl-2的表达降低较硼替佐米组快,Bax表达升高的亦快,且维C组24～48 h对Bax的表达增量远超过硼替佐米组.CCK-8检测发现,在72h时维C组、联合组、硼替佐米组Jurkat细胞的抑制率分别为25.72％、75.23％、56.81％;流式检测凋亡率在72 h分别为81.73％、67.06％与81.50％;维生素C组的早期凋亡率大于晚期凋亡率,而硼替佐米组与联合组均为早期凋亡率低,但维生素C的添加,联合用药组细胞的早期凋亡率得到了大约15.26％的提高.结论 维生素C抑制T细胞淋巴瘤细胞株Jurkat细胞的增殖,尤其是促进Jurkat细胞早期凋亡,且细胞凋亡调控蛋白Bax与Bcl-2参与了维生素C诱导Jurkat细胞凋亡机制.     

姜黄素诱导Jurkat细胞凋亡的机制

目的 探讨姜黄素诱导Jurkat细胞凋亡作用及凋亡机制.方法 体外培养Jurkat细胞,用姜黄素处理12 h后,MTT法检测细胞存活率;流式细胞学检测细胞凋亡率;Western Blot检测凋亡相关蛋白和内质网应激蛋白的表达变化.结果 姜黄素显著抑制Jurkat细胞存活率.随着姜黄素浓度增加,Jurkat细胞凋亡率增加,并从早期凋亡发展为晚期凋亡.姜黄素降低Bcl-2蛋白表达,增加Bax蛋白表达,增加GRP78蛋白表达,活化Caspase-3表达增高.结论 姜黄素启动内源性凋亡途径和内质网应激途径诱导Jurkat细胞发生凋亡.     

吉西他滨诱导膀胱癌T24细胞自噬的实验研究

目的 观察吉西他滨(gemcitabine,GEM)处理膀胱癌T24细胞后引起凋亡的同时是否诱导T24细胞自噬的产生,并初步探究其自噬产生的机制,以及自噬与凋亡之间的关系.方法 体外培养T24细胞,应用CCK-8法测定细胞抑制率;Western blot检测凋亡相关蛋白Caspase-3、PARP的活化,自噬标志蛋白LC3-Ⅱ、底物蛋白P62和自噬潮,以及JNK通路相关蛋白Jnk1、P-Jnk、Bcl-2的表达情况;并通过透射电镜观察细胞超微结构自噬体的变化;Annexin V-PI双流式细胞术检测抑制自噬后细胞凋亡率的变化情况.结果 CCK-8检测结果显示吉西他滨能有效抑制膀胱癌T24细胞增殖;1.0 μg/mL吉西他滨处理T24细胞4h即可发生明显的自噬,透射电镜观察可见自噬体小体的形成;Western blot检测结果显示吉西他滨作用T24细胞后cleaved-Caspase-3、PARP和LC3-Ⅱ表达增高,P62表达降低;P-Jnk出现活化,Bcl-2发生降解;而抑制自噬后能显著增强吉西他滨诱导的膀胱癌T24细胞的凋亡.结论 吉西他滨介导膀胱癌T24细胞凋亡的同时又诱导保护性自噬,JNK信号通路参与调控其自噬的发生,抑制自噬可以明显增强膀胱癌T24细胞对吉西他滨的化疗敏感性.     

马钱子碱对人白血病HL-60细胞Bcl-2与Bax基因表达的影响

目的:观察Bcl-2与Bax基因在马钱子碱诱导人白血病细胞株HL-60凋亡作用中的表达.方法:(1)用不同浓度马钱子碱溶液处理HL-60细胞,24、48、72 h后MTT法检测各组细胞的生长抑制率并计算IC50值;(2)用AO/EB染色法荧光显微镜下观察细胞的形态学变化;(3)用RT-PCR技术检测Bcl-2与Bax基因的表达.结果:马钱子碱可明显抑制HL-60细胞增殖并呈剂量和时间依赖性;以320 μg/mL马钱子碱作用48 h后大部分细胞核染色质呈桔红色,细胞核呈固缩状或圆珠状凋亡形态;不同浓度马钱子碱处理细胞后,其Bcl-2基因的表达随马钱子碱浓度的增加而降低,Bax基因的表达随马钱子碱浓度的升高而升高.结论:马钱子碱可明显抑制人白血病细胞株HL-60的增殖,诱导细胞凋亡,其机制可能与下调Bcl-2基因的表达和上调Bax基因的表达有关.     

穿山甲煎液诱导HL—60细胞凋亡的研究

目的：探讨中药穿山甲对急性髓细胞性白血病细胞株HL-60的作用。方法：以人白血病细胞株HL-60为研究对象。采用形态学观察。流式细胞仪、Caspase-3酶活性测定、免疫组化等方法。结果：穿山甲浓度在2.5-10mg/ml范围内可抑制HL-60细胞生长,并诱导其发生凋亡;并激活Caspase-3酶活性。下降bcl-2基因表达。结论：穿山甲对HL-60细胞有诱导凋亡作用。     

柔红霉素诱导急性淋巴细胞白血病Jurkat细胞凋亡的作用因素研究

目的 探讨柔红霉素(DNR)致急性淋巴细胞白血病Jurkat细胞株凋亡的作用因素.方法 不同浓度(0、0.05、0.1、0.5、1μg/mL)DNR及N-乙酰半胱氨酸(NAC)预处理组作用于Jurkat细胞株,采用MTT法检测细胞活力,Hoechst/PI双染法观察细胞凋亡,流式细胞术检测细胞活性氧(ROS)浓度及凋亡,RT-PCR法检测bax、survivin、bcl-2和bcl-xl基因的表达.结果 DNR可明显抑制Jurkat细胞株的活性,0、0.05、0.1、0.5、μg/mL的DNR及NAC预处理组分别作用于Jurkat细胞株24 h后,ROS水平分别为(7.98±0.55)%、(8.88±0.86)%、(9.46±0.98)%、(17.48±2.98)%、(24.46±2.43)%和(11.59±1.29)%,加入NAC后ROS生成受到抑制(P<0.01);细胞凋亡率分别为(11.41±1.44)%、(34.96±3.32)%、(45.58±3.12)%、(84.19±2.65)%、(87.93±1.74)%和(80.47±0.63)%,NAC预处理组与0.5 μg/mL DNR组比较,细胞凋亡率变化无统计学意义(P>0.05).NAC抑制bax基因mRNA表达,上调survivin、bcl-2和bcl-xl表达(均P<0.05).结论 DNR能够诱导Jurkat细胞株凋亡;ROS参与了DNR诱导的Jurkat细胞株凋亡;凋亡相关基因bax、bcl-2、bcl-xl及survivin能够与ROS相互作用,调节DNR诱导的Jurkat细胞株凋亡.     

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.