蛋白激酶C活性与电离辐射诱导人肝癌细胞株HepG2凋亡的关系

目的探讨蛋白激酶C(PKC)活性与X线电离辐射诱导人肝癌细胞株HepG2凋亡的关系.方法PKC调节剂PMA和SP预处理HepG2细胞,分辐射组、PMA0.5h组、PMA24h组、SP组;32P掺入法检测细胞的PKC活性,Varian2100直线加速器辐射细胞,流式细胞仪检测细胞凋亡率.结果6Gy X线辐射诱导HepG2细胞凋亡率是21.9%,辐射后细胞PKC活性是辐射前的2.79倍.辐射组、PMA0.5h组、PMA24h组、SP组的PKC活性分别为1.07、3.86、051、0.48 pmol/(min·μg),细胞凋亡率分别为21.9%、5.73%、31.70%和32.83%.结论辐射可以引起HepG2细胞凋亡,PKC活性增加/降低对辐射诱导的凋亡起抑制/促进作用,提示PKC可能参与了细胞凋亡保护机制.     

KBV200细胞多药耐药与蛋白激酶C亚型表达的关系

目的探讨蛋白激酶C(proteinkinaseC,PKC)亚型与KBV200肿瘤细胞多药耐药(multidrugresistance,MDR)的关系。方法(1)用Westernblotting法检测PKC亚型在耐药株KBV200及敏感株KB的表达及亚细胞分布;(2)流式细胞仪检测PKC亚型的荧光强度并对数据进行统计分析。结果(1)PKCα亚型的表达在KBV200中选择性升高,而PKCβ、ε无显著变化,PKCγ、ζ则未测出表达;(2)流式细胞仪检测发现KBV200细胞的PKCα的荧光强度较KB细胞显著升高,PKCβ、ε则无显著差别。结论PKC与KBV200MDR细胞株的MDR表型关系密切。在PKC亚型中,PKCα可能在KBV200细胞的MDR表型中起重要作用。     

他克莫司对癫痫持续状态大鼠海马PKCδ表达的影响

目的 研究氯化锂-匹鲁卡品癫痫持续状态(SE)大鼠模型中海马神经元蛋白激酶Cδ亚型(PKCδ)的表达及他克莫司(FK506)对其的影响.方法 将108只健康成年雄性大鼠随机分为对照组、癫痫组和FK506处理组,每组36只.采用HE染色法观察大鼠癫痫持续状态(SE)后海马神经元的损伤,应用逆转录PCR、免疫组织化学法检测各组大鼠海马PKCδ mRNA及蛋白的表达.结果 对照组PKCδ mRNA和蛋白仅有少量表达;与对照组相比,癫痫组各个时点PKCδ表达均显著增多(P＜0.05),SE后6 h PKCδ mRNA表达最高,PKCδ蛋白的平均光密度值SE后12h达高峰(P＜0.01),以后二者均逐渐降低,但72 h仍高于对照组水平(P＜0.05);FK506预处理可降低其表达,同时可减轻海马神经元损伤(P＜0.05).结论 FKS06可能通过抑制依赖PKCδ的细胞凋亡途径,发挥抗癫痫和脑保护作用.     

不同血液透析液对U937细胞凋亡和PKCδ表达的影响

目的 研究不同血液透析液对U937细胞蛋白激酶C-δ(PKCδ)表达及细胞凋亡的影响.方法 在对数生长期人单核细胞株U937细胞培养液中添加不同血液透析液进行干预,分为A液+B液组(血液透析液A液+B液组合)、A液+B液+PKCδ特异性抑制剂(rottlerin)组、A液+B粉组(血液透析液A液+B粉组合)、A液+B粉+ rottlerin组,同时设立空白对照和正常对照组.分组干预后24 h和48 h分别收集细胞,Hoechst33258染色后荧光显微镜观察细胞形态学改变;Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡;RT-PCR和Western blotting 检测PKCδ mRNA和蛋白表达.结果 A液+B粉组凋亡细胞明显增多,呈凋亡典型形态学改变,分组干预24 h和48 h的细胞凋亡率明显高于相应时间点空白对照组、正常对照组和A液+B粉+rottlerin组(P<0.05);同时与两对照组和A液+B粉+rottlerin组比较,A液+B粉组U937细胞PKCδ mRNA和蛋白表达均明显上调(P<0.05).A液+B液组细胞凋亡率及PKCδ mRNA和蛋白表达与两对照组和A液+B液+rottlerin组比较,差异均无统计学意义(P＞0.05).结论 血液透析液A液+B粉组合对U937细胞有明显促凋亡作用,其机制可能与上调细胞PKCδ表达有关;与A液+B粉组合比较,选择A液+B液组合作为血液透析液可降低患者外周血单核细胞的细胞凋亡率.     

不同血液透析液对U937细胞凋亡和PKCδ表达的影响

目的 研究不同血液透析液对U937细胞蛋白激酶C-δ(PKCδ)表达及细胞凋亡的影响.方法 在对数生长期人单核细胞株U937细胞培养液中添加不同血液透析液进行干预,分为A液+B液组(血液透析液A液+B液组合)、A液+B液+PKCδ特异性抑制剂(rottlerin)组、A液+B粉组(血液透析液A液+B粉组合)、A液+B粉+ rottlerin组,同时设立空白对照和正常对照组.分组干预后24 h和48 h分别收集细胞,Hoechst33258染色后荧光显微镜观察细胞形态学改变;Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡;RT-PCR和Western blotting 检测PKCδ mRNA和蛋白表达.结果 A液+B粉组凋亡细胞明显增多,呈凋亡典型形态学改变,分组干预24 h和48 h的细胞凋亡率明显高于相应时间点空白对照组、正常对照组和A液+B粉+rottlerin组(P<0.05);同时与两对照组和A液+B粉+rottlerin组比较,A液+B粉组U937细胞PKCδ mRNA和蛋白表达均明显上调(P<0.05).A液+B液组细胞凋亡率及PKCδ mRNA和蛋白表达与两对照组和A液+B液+rottlerin组比较,差异均无统计学意义(P＞0.05).结论 血液透析液A液+B粉组合对U937细胞有明显促凋亡作用,其机制可能与上调细胞PKCδ表达有关;与A液+B粉组合比较,选择A液+B液组合作为血液透析液可降低患者外周血单核细胞的细胞凋亡率.     

KBV2000细胞多药耐药蛋白激酶C亚型表达的关系

目的 探讨蛋白激酶C(protein kinaseC，PKC）亚型与KBV200肿瘤细胞多药耐药（multidrug resistance，MDR）的关系。方法 （1）用Western blotting法检测PKC亚型在耐药株KBV200及敏感株KB的表达及亚细胞分布；（2）流式细胞仪检测PKC亚型的荧光强度并对数据进行统计分析。结果 （1）PKCα亚型的表达在KBV200中选择性升高，而PKCβ、ε无显著变化，PKCγ、ζ则未测出表达；（2）流式细胞仪检测发现KBV200细胞的PKCα的荧光强度较KB细胞显著升高，PKCβ、ε则无显著差别。结论 PKC与KBV200MDR细胞株的MDR表型关系密切。在PKC亚型中，PKCα可能在KBV200细胞的MDR表型中起重要作用。     

石见穿多糖诱导肝癌细胞凋亡的实验研究

目的探讨石见穿多糖诱导人肝癌HepG2细胞凋亡作用。方法荧光显微镜观察细胞凋亡的形态学改变;利用流式细胞仪Annexin V-FITC/PI法检测经不同浓度石见穿多糖作用的肝癌HepG2细胞的凋亡率。结果荧光显微镜观察可见HepG细胞不同时期凋亡形态学改变;石见穿多糖各浓度组平均细胞凋亡率分别为25mg/L组(8.27±1.84)%、50mg/L组(13.53±2.67)%、100 mg/L组(22.03±2.48)%,显著高于对照组(0.57±0.23)%(P0.01)。结论石见穿多糖能够诱导肝癌细胞HepG2凋亡,且凋亡率随着石见穿多糖浓度的增加而增高。     

双歧杆菌DNA对巨噬细胞MAPK和PKG以及NF-kB的影响

目的 以信号分子MAPK家系、PKC家族和NF-kB为线索探索青春型双歧杆菌的DNA激活巨噬细胞的途径.方法 以激光共聚焦显微镜定量测定小鼠腹腔巨噬细胞ERK1/2、JNK、p38、PKCα、PKCβⅠ、PKCβⅡ、PKCγ、PKC ε和PKCζ的含量.以细胞免疫化学方法检测巨噬细胞NF-kB的阳性细胞密度.结果 双歧杆菌DNA注射组小鼠腹腔巨噬细胞ERK1/2、PKCα和PKCβH的平均荧光强度明显高于对照组(P＜0.01),而JNK、p38、PKCβⅠ、PKCγ、PKC ε和PKCζ的平均荧光强度在两组间则差异无显著性(P＞0.05).双歧杆菌DNA注射组巨噬细胞NF-kB的阳性细胞密度显著高于对照组(P＜0.01).结论 青春型双歧杆菌的DNA可通过活化ERK1/2、PKC α、PKC βⅡ和NF-kB来激活巨噬细胞.     

不同强度+Gz暴露对大鼠心肌细胞凋亡的影响

目的: 探讨不同强度的持续性正加速度( Gz)对大鼠心肌细胞凋亡的影响. 方法: 12只SD大鼠随机分为对照组、 6Gz暴露组、 10 Gz暴露组,每组4只, Gz暴露组分别给予 6Gz和 10Gz负荷刺激3 min,于暴露后6 h取大鼠心肌组织,用原位末端标记法(TUNEL)检测心肌细胞凋亡情况并计数. 结果: 对照组大鼠偶见TUNEL阳性染色的心肌凋亡细胞; 6Gz暴露组大鼠心肌凋亡细胞数较对照组显著增多(P＜0.05); 10Gz暴露组大鼠心肌凋亡阳性细胞数较对照组及 6 Gz组均明显增多(P＜0.05). 结论: Gz暴露可以引起大鼠心肌细胞凋亡,且随着G值的增大,心肌凋亡细胞数目逐渐增多.     

一氧化碳调节低氧大鼠肺动脉平滑肌细胞凋亡

目的: 探讨一氧化碳对低氧大鼠肺血管平滑肌细胞凋亡的影响及其机制.方法:将大鼠随机分为4组:常氧对照组(n=7)、低氧组(n=6)、低氧+锌原卟啉组(低氧+ ZnPP组,n=6)及低氧+低浓度一氧化碳组(低氧+低浓度CO组,n=5).应用Bcl-2、Caspase-3多克隆抗体和NF-κΒ单克隆抗体经免疫组织化学检测技术及原位缺口末端标记法(TUNEL)对大鼠肺动脉平滑肌细胞进行凋亡定位及半定量分析.结果: (1)低氧组大鼠肺动脉平滑肌凋亡细胞百分数和平滑肌细胞Caspase-3表达积分低于对照组,NF-κΒ阳性细胞百分数和平滑肌细胞Bcl-2表达积分明显高于对照组;(2)低氧+ ZnPP组大鼠肺动脉平滑肌凋亡细胞百分数明显低于对照组和低氧组,平滑肌细胞Caspase-3表达积分明显低于对照组,与低氧组无差异;NF-κΒ阳性细胞百分数和平滑肌细胞Bcl-2表达积分明显高于对照组和低氧组;(3)低氧+低浓度CO组大鼠肺动脉平滑肌凋亡细胞百分数和平滑肌细胞Caspase-3表达积分明显高于对照组、低氧组及低氧+ZnPP组,NF-κΒ阳性细胞百分数和平滑肌细胞Bcl-2表达积分明显低于低氧组及低氧+ZnPP组,与对照组差异无显著性.结论: CO促进低氧大鼠肺动脉平滑肌细胞凋亡,同时还伴有Bcl-2和NF-κΒ的变化.     

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.