IL-22抑制ox-LDL诱导的CRL-1730细胞凋亡并上调其Bcl-2表达

目的研究IL-22对ox-LDL诱导的CRL-1730细胞(人脐静脉内皮细胞)凋亡的影响及其机制。方法 100μg/ml ox-LDL刺激CRL-1730细胞,不同时间点经实时荧光定量RT-PCR检测CRL-1730细胞中IL-22受体R1(IL-22R1)mRNA的表达状况。然后在ox-LDL诱导的CRL-1730细胞凋亡模型中加入20 ng/ml外源性IL-22,通过AnnexinⅤ-PI凋亡试剂盒检测其对细胞凋亡的影响,经实时荧光定量反转录PCR检测Bcl-2、Bcl-xl、Bcl-w、caspase 3及STAT3mRNA的表达,采用免疫印迹法分析STAT3磷酸化和Bcl-2蛋白的水平。结果 IL-22R1 mRNA在ox-LDL刺激后表达明显增高(P<0.05),且于12 h达峰值(P<0.01);加入IL-22可明显抑制ox-LDL刺激诱导的内皮细胞凋亡(P<0.01),促进抗凋亡基因Bcl-2、Bcl-xl和Bcl-w表达,而降低caspase 3 mRNA表达(P<0.01)。此外,IL-22作用1～2 h STAT3发生磷酸化,4 h后Bcl-2蛋白表达升高(P<0.01)。结论 IL-22可抑制ox-LDL诱导的CRL-1730细胞凋亡,这可能与其促使STAT3磷酸化,上调抗凋亡分子Bcl-2、Bcl-xl和Bcl-w的表达,抑制促凋亡分子caspase 3的表达有关。     

MiR-590-5p对ox-LDL诱导血管内皮细胞凋亡以及LOX-1表达的影响

目的:研究miR-590-5p对氧化低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVECs)凋亡过程中血凝素样氧化低密度脂蛋白受体1(LOX-1)表达的调节作用,观察miR-590-5p对HUVECs凋亡的影响。方法:HUVECs经ox-LDL(50μg/mL)作用不同时间(0~48 h)。Annexin V-FITC/PI双染流式细胞仪(FCM)检测细胞凋亡率,实时荧光定量PCR(RT-qPCR)检测miR-590-5p的表达变化。HUVECs转染miR-590-5p模拟物(miR-590-5pmimics)后ox-LDL继续作用48 h。RT-qPCR和Western印迹分别检测LOX-1 mRNA和蛋白的表达情况。FCM检测HUVECs凋亡率的变化。结果:随ox-LDL作用时间的延长,HUVECs凋亡率逐渐升高,miR-590-5p表达逐渐下调。miR-590-5p模拟物转染后可以降低LOX-1 mRNA和蛋白的表达,并抑制ox-LDL诱导的HUVECs凋亡。结论:miR-590-5p可抑制ox-LDL诱导的HUVECs凋亡,其机制可能与下调LOX-1基因表达有关。     

积雪草苷通过Vimentin/NLRP3通路诱导鼻咽癌细胞凋亡研究

目的研究积雪草苷对人鼻咽癌CNE-2细胞生长、凋亡及凋亡通路的影响。方法体外培养CNE-2细胞,将不同浓度的积雪草苷作用于CNE-2细胞,MTT和CCK-8法检测积雪草苷对细胞生长的影响;流式细胞术检测对细胞凋亡的影响;Western blotting检测Vimentin、NLRP3、Bcl-2和Caspase-3表达;ELISA法检测TNF-α和IL-1β含量。结果MTT和CCK-8结果显示,积雪草苷可浓度依赖性地抑制CNE-2细胞的生长;流式细胞仪检测结果显示,积雪草苷可浓度依赖性地促进CNE-2细胞凋亡;Western blotting结果显示,积雪草苷促进Caspase-3蛋白的表达,抑制Vimentin、NLRP3和Bcl-2的表达;ELISA法结果显示,积雪草苷可浓度依赖性地降低TNF-α和IL-1β含量。结论积雪草苷可显著促进人鼻咽癌细胞株CNE-2细胞凋亡,其机制可能与下调Vimentin,NLRP3蛋白的表达来实现的。     

淫羊藿苷对ox-LDL诱导的RAW264.7细胞凋亡及炎症的影响

目的 研究淫羊藿苷(ICA)对氧化低密度脂蛋白(ox-LDL)诱导的小鼠单核巨噬细胞白血病细胞(RAW264.7)凋亡及炎症的影响,进一步阐明ICA抗动脉粥样硬化(AS)的作用。方法 体外培养RAW264.7细胞,ox-LDL诱导RAW264.7细胞构建泡沫细胞模型,观察ICA低、中、高剂量对RAW264.7细胞活力、细胞凋亡、泡化沫、炎症因子IL-1β、IL-6和TNF-α分泌的影响,同时采用Western blot检测ICA对Bcl-2、Bax、Caspase-3、IκBα、NF-κB p65蛋白表达的影响。结果 ICA可以提高细胞活力,降低细胞的凋亡率;减少泡沫巨噬细胞的形成;降低炎症因子IL-1β、IL-6和TNF-α的分泌;下调Bcl-2、Caspase-3、NF-κB p65蛋白表达,上调Bax、IκBα的蛋白表达。结论 ICA能够减轻泡沫巨噬细胞的凋亡,抑制炎症因子的表达,从而可能延缓动脉粥样硬化的发展进程。     

球状脂联素对晚期糖基化终产物诱导人脐静脉内皮细胞凋亡的蛋白表达影响

目的:研究球状脂联素(gAd)抑制晚期糖基化终产物(AGEs)诱导人脐静脉内皮细胞(HUVECs)凋亡的相关蛋白表达。方法:体外用AGEs培养HUVECs,经四甲基偶氮唑盐比色法(MTT)测定细胞活性,Annexin V-FITC/PI双染标记法检测细胞凋亡,免疫印记法检测Bax、Bcl-2蛋白的表达。结果:AGEs作用HUVECs,经MTT和流式细胞仪检测,细胞凋亡具有AGEs浓度依赖性。相同浓度AGEs作用HUVECs时,gAd预处理组与非预处理组比较,明显抑制HUVECs凋亡。免疫印记结果显示gAd预处理组与非预处理组比较,gAd可明显抑制Bax蛋白表达,增强Bcl-2蛋白表达。结论:gAd可通过激活Bcl-2、抑制Bax蛋白表达,拮抗AGEs诱导HUVECs凋亡。     

PM2.5对大鼠肺泡巨噬细胞内NLRP3炎性小体表达及细胞凋亡的影响

目的:初步探讨大气细颗粒物(fine particulate matter,PM2.5)诱导大鼠肺泡巨噬细胞内NLRP3炎性小体活化及细胞凋亡的潜在分子机制.方法:采用中流量采样器收集PM2.5颗粒物,经超声提取制成混悬液,作用于大鼠肺泡巨噬细胞(NR8383)后,MTT法检测细胞存活率,Western blot检测Nod样受体蛋白3(NLRP3)、组织蛋白酶B(Cathepsin-B)及凋亡相关蛋白[Bax、Bcl-2及半胱天冬酶-3(Caspase-3)]的表达水平,ELISA检测PM2.5诱导的细胞上清液中半胱天冬酶-1(Caspase-1)、白介素(in-terleukin,IL)-18和IL-1β的含量.结果:随着PM2.5浓度的增加和刺激时间的延长,NR8383细胞存活率降低(24 h:F=17.253,P=0.000;48 h:F=18.678,P=0.000;72 h:F=256.104,P=-0.000);PM2.5诱导NR8383细胞内NLRP3 (F=437.166,P=0.000)和Cathep-sin-B(F=42.062,P=0.000)蛋白的表达上调;PM2.5上调细胞内促凋亡蛋白Bax(F=72.827,P=0.000)表达,下调抗凋亡蛋白Bcl-2(F=390.322,P=0.000)表达,下调Caspase-3(F=169.833,P=-0.000)蛋白的表达;PM2.5上调细胞培养液中Caspase-1(F=629.866,P=0.000)、IL-18(F=587.165,P=0.000)和IL-1β(F=68.472,P=0.000)的水平.结论:PM2.5可通过溶酶体模式诱导肺泡巨噬细胞内NLRP3炎性小体活化,促进IL-18和IL-1β分泌,并引起肺泡巨噬细胞发生凋亡.     

银杏黄酮苷元对氧化低密度脂蛋白所致内皮细胞功能的影响

目的探讨银杏黄酮苷元(GA)对氧化低密度脂蛋白(ox-LDL)诱导脐静脉内皮细胞株ECV304单核细胞趋化蛋白-1(MCP-1)和植物血凝素样低密度脂蛋白受体-1(LOX-1)表达的调节作用。方法应用逆转录-聚合酶链式反应(RT-PCR)、酶联免疫吸附反应(ELISA)、SP免疫组化法等,探讨ox-LDL诱导下内皮细胞表达MCP-1、LOX-1及银杏黄酮苷元的干预作用。结果6.25～25 mg/L银杏黄酮苷元与脐静脉内皮细胞株ECV304共同培养6～48 h可显著抑制ox-LDL诱导的内皮细胞MCP-1、LOX-1 mRNA和蛋白表达(P0.05),具有浓度、时间效应关系(P0.05)。LOX-1拮抗剂PIA可抑制ox-LDL诱导的内皮细胞LOX-1、MCP-1 mRNA和蛋白表达(P0.05)。结论银杏黄酮苷元可显著抑制ox-LDL诱导的内皮细胞黏附因子MCP-1表达,该作用可能通过抑制LOX-1的表达来实现。     

苯扎贝特对ox-LDL诱导内皮细胞凋亡基因Bcl-2/Bax的影响

目的 观察苯扎贝特对氧化型低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞凋亡基因Bcl-2/Bax的影响. 方法 体外培养人脐静脉内皮细胞(HUVECs),根据实验要求分为正常对照组、ox-LDL组、低浓度苯扎贝特(50μmol/L)组、中浓度苯扎贝特(100 μmol/L)组、高浓度苯扎贝特(200μmol/L)组.RT-PCR观察各组凋亡基因Bcl-2、Bax及Bd-2/BaxmRNA的变化. 结果 与正常对照组比较,ox-LDL组凋亡基因Bcl-2表达下降(P<0.05),凋亡基因Bax表达增加(P<0.05),Bcl-2/Bax比值下降(P<0.05);不同浓度苯扎贝特组与ox-LDL组比较,Bcl-2表达增加(P<0.05),Bax表达降低(P<0.05),Bcl-2/Bax上调(P<0.05),且呈浓度效应依赖关系. 结论 ox-LDL可引起内皮细胞抗凋亡基因Bcl-2表达下调,凋亡基因Bax表达上调,Bcl-2和Bax比值下降,从而引起内皮细胞凋亡增加;苯扎贝特可通过上调Bcl-2与Bax的比值抑制ox-LDL引起的内皮细胞凋亡,起到抗动脉粥样硬化作用.     

积雪草苷调控基质金属蛋白酶-9减轻川崎病血管损伤的机制研究

目的 探讨积雪草苷对川崎病血管损伤的影响及其可能作用机制。方法 采用肿瘤坏死因子-α(TNF-α)诱导人冠状动脉血管内皮细胞建立川崎病血管损伤模型,分为7组：con组、TNF-α组、TNF-α+低积雪草苷组、TNF-α+中积雪草苷组、TNF-α+高积雪草苷组、TNF-α+积雪草苷+pcDNA组、TNF-α+积雪草苷+pcDNA-MMP-9组;CCK-8法、流式细胞术分别检测细胞增殖及凋亡;q RT-PCR检测基质金属蛋白酶-9 (MMP-9) m RNA的表达水平;ELISA法检测IL-6、IL-1的水平;Western blot法检测Bax、Bcl-2、MMP-9蛋白表达量。结果 与con组比较,TNF-α组细胞凋亡率和Bax蛋白水平升高,IL-6、IL-1的水平升高,MMP-9 m RNA及蛋白水平升高,Bcl-2蛋白水平降低,差异均有统计学意义(P<0.05);与TNF-α组比较,TNF-α+低积雪草苷组、TNF-α+中积雪草苷组、TNF-α+高积雪草苷组细胞凋亡率和Bax蛋白水平降低,IL-6、IL-1的水平降低,MMP-9 m RNA及蛋白水平降低,Bcl-2蛋白水平升...     

NLRP3炎症小体及其下游炎症因子在动脉粥样硬化炎症反应中的作用

目的：探讨NLRP3炎症小体及其下游炎症因子在动脉粥样硬化炎症反应中的作用。方法选择氧化低密度脂蛋白(ox-LDL)作用于人单核细胞系THP-1,经Western blot法检测NLRP3及其下游炎症因子半胱天冬酶-1(Caspase-1)、白介素-1β(IL-1β)和白介素-18(IL-18)的表达;并采用SiRNA技术下调空白处理组(Blank组)和ox-LDL处理组NLRP3基因表达,分别检测以上指标。结果与空白处理组相比,50 mg/L、100 mg/L、150 mg/L的ox-LDL作用于单核细胞后可促进NLRP3的表达,NLRP3下游的Caspase-1、IL-1β和IL-18表达水平与NLRP3表达一致,表达也均明显升高,差异均具有统计学意义(P<0.05)。NLRP3小干扰RNA (SC-45469)转染后,Blank组与ox-LDL组的NLRP3及其下游炎症因子Caspase-1、IL-1β和IL-18的表达均有显著下降(P<0.05);而转染后Blank组与ox-LDL组各指标下降率组间比较差异均无统计学意义(P>0.05)。结论 NLRP3炎症小体参与动脉粥样硬化发生发展的过程,其可能通过改变ox-LDL的表达水平,诱发炎症相关因子Caspase-1、IL-1β和IL-18的分泌来调节, NLRP3炎症小体具有成为临床上防治动脉粥样硬化的新靶点的重要潜力。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.