紫外线对HaCaT细胞损伤作用的研究

目的 探讨中波紫外线(UVB)对人永生化角质形成细胞(HaCaT细胞)的损伤影响,建造UVB损伤人表皮细胞的模型,以方便进行实验室光损伤研究.方法 取培养的HaCaT细胞,用10、30、50、70、90 mJ/cm 的UVB照射后继续培养24 h,在光学显微镜下观察细胞的形态学改变,以MTT法检测细胞的增殖活性,以ELISE法测定细胞上清液中TNF-α的水平,以流式细胞术检测细胞凋亡率.结果 随UVB照射剂量的增加,在光镜下可见HaCaT细胞不同程度的损伤,其增殖活性降低,P＜0.01;细胞上清液中TNF-α的水平增加,P＜0.01;凋亡率增加,P＜0.01.结论 UVB可损伤HaCaT细胞,降低增殖活性,增加细胞上清液中TNF-α的水平,诱导细胞凋亡,并且呈剂量依赖性.在实验中,可根据需要调节不同的UVB照射剂量,建造所需的细胞损伤模型.     

竹叶总黄酮对UVB诱导HaCaT细胞氧化损伤的保护作用

目的?研究了竹叶总黄酮对紫外线B(UVB)诱发HaCaT角质细胞氧化损伤的保护效果和机制。方法?HaCaT细胞经竹叶总黄酮预处理24?h后,以辐射剂量为20?mJ/cm2的UVB照射细胞2?h后,MTT法测定细胞生存率。同时测定细胞内活性氧(ROS)水准,脂质过氧化程度,抗氧化酶(超氧化物歧化酶,SOD;过氧化氢酶,CAT和过氧化物歧化酶,GSH-Px)并利用RT-PCR法分析其mRNA转录水准。ELISA法分析TNF-α和IL-6分泌量。结果?竹叶总黄酮可有效地抑制UVB导致的细胞氧化损伤并提高其细胞生存率。与对照组相比,竹叶总黄酮还能有效降低受损细胞内ROS水准,脂质过氧化程度,并提高SOD、CAT和GSH-Px等3种内源性抗氧化酶的活性及其它们的mRNA转录水平。此外,竹叶总黄酮还能抑制UVB所造成受损细胞分泌TNF-α和IL-6炎性细胞因子的能力。结论?竹叶总黄酮可通过降低细胞内ROS水平,抑制细胞脂质过氧化,调节细胞内氧化酶的活性抑制UVB导致的HaCaT细胞氧化损伤。此外,竹叶总黄酮还可降低UVB所诱发的TNF-α和IL-6分泌。      

黄芩对皮肤细胞紫外线辐射损伤的保护作用

目的 研究传统中药黄芩对紫外线(UVA、UVB)损伤皮肤细胞(角质形成细胞和成纤维细胞)的保护作用。方法 采用30、60、90ml／cm^2的UVB和4、8、12J／cm^2的UVA照射培养的原代角质形成细胞和成纤维细胞，加入中药黄芩进行干预处理，以MTT法检测细胞活性。结果 UVA、UVB照射可引起皮肤角质形成细胞和成纤维细胞损伤，而黄芩预处理后细胞活性可恢复8％～38％。结论 黄芩具有光保护性能，可减轻UVA、UVB对皮肤细胞的损伤作用。     

盐酸13-己基小檗碱对T淋巴细胞活化和角质形成细胞细胞因子产生的影响

目的研究盐酸13-己基小檗碱对T淋巴细胞活化、角质形成细胞增殖、以及白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)和白介素-1(IL-1)产生的影响,探索该化合物针对炎症性皮肤病的作用靶点。方法体外培养小鼠脾脏T淋巴细胞、永生化角质形成细胞株HaCaT细胞以及小鼠腹腔巨噬细胞,分别用伴刀豆球蛋白A(ConA)、重组人肿瘤坏死因子-α(rhTNF-α)、312nm窄波UVB和脂多糖(LPS)诱导,采用MTT法检测盐酸13-己基小檗碱对小鼠T淋巴细胞活化和HaCaT细胞增殖的影响,用ELISA法检测其对HaCaT细胞IL-8和TNF-α,以及小鼠腹腔巨噬细胞IL-1β产生的影响。结果盐酸13-己基小檗碱明显抑制小鼠T淋巴细胞活化(IC50≈0.123μg/ml);在0.05～0.78μg/ml范围对HaCaT细胞增殖无明显的直接影响(抑制率<15%),能剂量依赖性抑制25ng/ml rhTNF-α诱导的HaCaT细胞IL-8产生(IC50=0.13μg/ml)和124.2 mJ/cm2窄波UVB照射引起的HaCaT细胞TNF-α产生(IC50=0.21μg/ml);在0.01μg/ml浓度时抑制10μg/ml LPS诱导的小鼠腹腔巨噬细胞IL-1β产生(抑制率达50%)。结论盐酸13-己基小檗碱能抑制T淋巴细胞活化及炎症性细胞因子IL-8、TNF-α和IL-1β产生,提示以上作用可能是该化合物针对炎症性皮肤病的作用靶点。     

枸杞多糖在UVB辐射人角质形成细胞氧化损伤中的保护作用

目的利用中波紫外线辐射体外培养的人角质形成细胞（HaCaT）建立紫外线损伤模型,探讨枸杞多糖（LBP）在紫外线辐射HaCaT氧化损伤中的作用及其机制,为抗氧化剂的研发提供理论依据。方法在HaCaT培养的基础上设置空白对照组、UVB照射组和不同浓度的LBP干预组,采用UVB辐射HaCaT进行造模,运用酶生化法检测不同浓度的枸杞多糖对UVB辐射HaCaT后的细胞增殖及抗氧化酶的影响。结果 UVB辐射对HaCaT造成明显损伤,LBP可提高UVB辐射后HaCaT细胞增值活性（MTT）,提高超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性,降低丙二醛（MDA）产生。（P〈0.05）。结论 UVB对HaCaT细胞有损伤作用,LBP可拮抗UVB所致HaCaT细胞抗氧化酶活性的降低,从而具有抗氧化光保护作用。     

瑞香素对TNF-α刺激的HaCaT角质形成细胞炎症因子表达的影响

目的观察瑞香素对TNF-α刺激的HaCaT角质形成细胞炎症因子表达的影响,并初步探讨其机制。方法细胞活力检测采用CCK-8检测试剂盒。实时荧光定量PCR检测炎症因子IL-1β、IL-6、TNF-α、IL-8和MCP-1 mRNA的水平。Western blotting检测p65和p-p65的蛋白水平变化。结果 20μM以下瑞香素处理HaCaT角质形成细胞24小时未表现出明显的细胞毒性。TNF-α刺激能显著促进HaCaT角质形成细胞IL-1β(P0.05)、IL-6(P0.001)、TNF-α(P0.05)、IL-8(P0.05)和MCP-1(P0.05)的表达;而瑞香素(20μM)联合处理能减弱TNF-α刺激所引起的炎症因子表达升高。TNF-α处理能够诱导p65磷酸化,而瑞香素(20μM)联合处理能够抑制p65的磷酸化。结论瑞香素对TNF-α诱导的HaCaT角质形成细胞炎症反应具有抑制作用,而这种抑制作用与NF-κB信号通路的抑制相关。     

参芩仿生化提取物对中波紫外线照射致生物体损伤的保护作用

目的通过人参、黄芩仿生化提取物,对中波紫外线(UVB)照射致人永生化角质形成细胞(HaCaT)及小鼠皮肤损伤的保护作用进行研究,为中药抗辐射提供理论依据。方法采用MTT法检测HaCaT细胞生存率,黄嘌呤氧化酶法检测细胞溶解液中及小鼠皮肤组织匀浆中超氧化物歧化酶(SOD)活性,硫代巴比妥酸法检测细胞溶解液中及小鼠皮肤组织匀浆中丙二醛(MDA)含量变化。结果参芩仿生化提取物能够显著提高UVB照射损伤细胞的活性,明显升高细胞溶解液及小鼠皮肤组织匀浆中SOD活力,降低MDA含量。结论参芩仿生化提取物对UVB辐射致HaCaT细胞和小鼠皮肤损伤均有明显的保护作用,能够有效的抵抗UVB所致的氧化损伤。     

光老化人角质形成细胞模型的构建

目的:探讨中波紫外线(UVB)辐射导致人角质形成细胞(HaCaT细胞)光老化的模型构建及机制研究.方法:体外培养HaCaT细胞分为空白组和实验组,模型组细胞用不同照射剂量的UVB照射,空白组细胞不予紫外线照射,采用CCK-8法检测细胞相对活力,筛选出最适合的照射剂量用于建立光老化模型;应用同样的方法体外培养HaCaT细...     

羟氯喹及中药对紫外线损伤角质形成细胞保护作用的实验研究

目的 :研究羟氯喹及某些中药活性成分对紫外线损伤角质形成细胞保护作用的影响及作用环节。方法 :采用 30、6 0、90mJ cm2 剂量的UVB照射培养的永生的角质形成细胞HaCaT ,以羟氯喹及没食子儿茶素没食子酸脂(EGCG)、黄芩、川芎进行干预处理 ,观察其保护性能及作用环节。以普通光学显微镜对照观察细胞受损程度 ,记录 72h内细胞生长曲线 ,以MTT法检测细胞活性 ,以酶联免疫吸附实验检测IL 6和TNF α的分泌量。结果 :HaCaT细胞经UVB照射后 ,受试角质形成细胞的损伤程度与紫外线照射剂量呈正相关 ,细胞计数与活性下降 39%～ 80 % ,在照射后72h损伤程度超过 80 %。药物处理后细胞活性可恢复10 %～ 72 %。经比较表明 ,羟氯喹具有一定的光保护作用(OD值为 [0 4 3± 0 0 4 ]至 [0 96± 0 0 4 ],P <0 0 5 )。EGCG具有较强的光保护效应 (OD值为 [1 19± 0 0 7]至 [1 2 8±0 0 6 ],P <0 0 1) ,空白组为 ([0 2 6± 0 0 4 ]～ [0 79± 0 0 2 ])。此外 ,EGCG可抑制角质形成细胞释放IL 6和TNF 等细胞因子 ;羟氯喹及黄芩只能抑制IL 6的释放 ,对TNF α无明显抑制作用 ;而川芎则未见有明显抑制角质形成细胞释放IL 6和TNF α等细胞因子的能力。结论 :UVB对皮肤细胞的损伤作用具有剂量递增性 ;羟氯喹及茶多酚 (EGC     

UCP2对紫外线诱导细胞凋亡的影响

研究表明,紫外线辐射诱导细胞内活性氧(ROS)过度累积是导致细胞氧化损伤的最直接因素之一。长波紫外线(UVA)或中波紫外线(UVB)照射角质形成细胞后均可引起DNA损伤,UVA主要是通过生成ROS间接损伤DNA,UVB则是直接损伤DNA[1-2]。线粒体是细胞中的膜性结     

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.