心脑舒通及其单体成分对内毒素激活的小胶质细胞的影响

[目的]研究心脑舒通及其单体成分对小胶质细胞(BV-2)的影响及其作用机制。[方法]实验分为对照组、脂多糖刺激组、加药组,检测心脑舒通(不同稀释倍数的心脑舒通及其单体成分)对小胶质细胞活力及脂多糖(LPS)诱导小胶质细胞一氧化氮(NO)产生的影响。[结果]心脑舒通对LPS诱导的小胶质细胞NO的产生无明显抑制作用,在心脑舒通13种单体成分中,伪原薯蓣皂苷、支脱皂苷元、薯蓣皂苷元可以在不影响细胞活力的情况下抑制NO产生。[结论]心脑舒通部分单体成分抑制激活小胶质细胞NO的产生可能是心脑舒通发挥神经保护作用机制之一。     

西洋参不同部位皂苷成分研究

[目的] 研究西洋参不同部位皂苷类成分的含量差异。[方法] 对西洋参9个部位以甲醇为溶剂分别进行回流提取,用HPLC法测定西洋参不同部位中7种单体皂苷,即人参皂苷Rg1、Re、Rb1、Rc、Rb2、Rb3和Rd的含量,并对测定结果分别进行主成分分析和聚类分析。[结果] 西洋参花蕾、须根、芦头中7种皂苷含量较高,用主成分分析和聚类分析将9个部位分成两类,花蕾和叶为一类,其他部位为另一类。[结论] 西洋参各部位总皂苷及单体皂苷含量差异较大,通过HPLC对其进行含量测定,并结合主成分和聚类分析,可对各部位皂苷成分的分布及含量进行全面评价。     

白藜芦醇抑制TXNIP/NLRP3信号通路改善脂多糖诱导的肾小管上皮细胞炎性损伤

[目的]观察白藜芦醇对脂多糖诱导的肾小管上皮细胞（HK-2）炎性损伤的保护作用,及其对TXNIP/NLRP3炎性通路的影响。[方法]采用脂多糖（LPS,1 mg/L）体外诱导HK-2细胞炎性损伤模型,将细胞分为正常对照组、LPS诱导组、LPS+低剂量（50 μmol/L）白藜芦醇组、LPS+中剂量（100 μmol/L）白藜芦醇组、LPS+高剂量（200 μmol/L）白藜芦醇组;CCK8法检测HK-2细胞相对存活率,酶联免疫吸附测定（ELISA）法检测炎性因子白介素-6（IL-6）、白介素-1β（IL-1β）及肿瘤坏死因子-α（TNF-α）水平,Western Blot方法分析诱导型一氧化氮合酶（iNOS）、环氧酶-2（COX-2）、人硫氧还蛋白互作蛋白（TXNIP）及人隐热蛋白（NLRP3）表达。[结果]与LPS诱导组比较,不同浓度白藜芦醇干预的HK-2细胞相对存活率明显升高（P<0.05）,细胞上清炎性因子IL-6、IL-1β及TNF-α水平明显降低（P<0.05）,细胞中炎性蛋白iNOS及COX-2表达水平显著降低（P<0.05）;细胞中TXNIP及NLRP3蛋白表达也显著降低（P<0.05）,并表现出剂量依赖性。[结论]白藜芦醇可以明显抑制LPS诱导的HK-2细胞炎症损伤,抑制炎症相关蛋白的表达,其作用机制可能与抑制TXNIP/NLRP3炎性通路活化相关。     

灯盏乙素对LPS诱导的RAW264.7细胞炎症介质分泌的影响

[目的] 研究灯盏乙素对脂多糖（LPS）诱导的RAW264.7细胞炎症的影响,初步探讨其抗炎作用机制。[方法] 用噻唑蓝（MTT）法检测灯盏乙素对RAW264.7细胞活力的影响;用Griess法检测灯盏乙素对LPS诱导的RAW264.7细胞中一氧化氮（NO）生成的影响;用荧光素酶报告质粒法检测灯盏乙素对LPS诱导的核转录因子（NF-κB）转录活性的影响;用实时定量聚合酶链反应（PCR）法检测灯盏乙素对LPS诱导的RAW264.7细胞中NF-κB靶基因肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）、白介素-6（IL-6）mRNA表达的影响。[结果] 0.01～10 μmol/L的灯盏乙素处理24 h后对RAW264.7活力无显著影响;1、10 μmol/L的灯盏乙素可以显著抑制LPS诱导的NO的生成及NF-κB转录活性;灯盏乙素可以不同程度的抑制LPS诱导的RAW264.7细胞NF-κB靶基因TNF-α、IL-1β、IL-6 mRNA的上调作用。[结论] 灯盏乙素可能通过抑制NF-κB的活化及炎症因子的表达发挥一定的抗炎作用。     

高效液相色谱法测定人参皂苷Rg1与黄芩苷在细胞培养基中的含量变化

[目的]研究细胞培养基中人参皂苷Rg1和黄芩苷稳定性问题。[方法]采用高效液相色谱法测定细胞培养基中人参皂苷Rg1和黄芩苷的含量。[结果]人参皂苷Rg1和黄芩苷的线性范围均为20～400ng,方法回收率分别为96.8%和97.7%。[结论]该法简便、灵敏、特异,适用于细胞培养基中人参皂苷Rg1和黄芩苷的测定。     

妍婷颗粒对脂多糖诱导小鼠脾细胞产生TNF-α的影响

[目的]研究中药复方妍婷颗粒对脂多糖(LPS)诱导小鼠脾细胞产生肿瘤坏死因子-α(TNF-α)的影响,初步探讨妍婷颗粒对慢性盆腔炎抗炎、免疫调节的内在机制。[方法]体外培养小鼠脾细胞用LPS刺激,用不同浓度妍婷颗粒含药血清干预,酶联免疫银光法(ELISA)法检测上清液中TNF-ɑ含量,反向逆转录聚合酶链式反应(RT-PCR)法测细胞中TNF-ɑmRNA表达。[结果]1)经LPS诱导后小鼠脾细胞TNF-ɑ分泌量明显升高,TNF-ɑmRNA表达增强。2)小鼠脾细胞TNF-ɑ的分泌量与表达与妍婷颗粒含药血清浓度呈剂量依赖性。[结论]妍婷颗粒具有抑制LPS诱导的小鼠脾细胞产生TNF-ɑ的作用,据此推测妍婷颗粒的抗炎、免疫调节作用机制可能与其抑制脂多糖介导的TNF-ɑ释放有关。     

清感冬饮通过调控NF-κB/iNOS/NO信号通路抑制LPS诱导的RAW264.7巨噬细胞炎症

[目的] 通过构建脂多糖（LPS）诱导的RAW264.7巨噬细胞炎症模型,探讨清感冬饮（QGDY）的抗炎作用及其机制。[方法] 采用CCK-8法和LDH法检测不同浓度的清感冬饮对HEK293T细胞和RAW264.7巨噬细胞的细胞活力及LDH漏出量的影响。分别建立抗氧化反应元件（ARE）和核因子-κB（NF-κB）双荧光素酶报告系统,考察清感冬饮对HEK293T细胞ARE、NF-κB表达情况的影响。建立LPS诱导的RAW264.7巨噬细胞体外炎症模型,将RAW264.7巨噬细胞随机分为空白对照组、LPS模型组、清感冬饮低、中、高剂量组（1、10、100μg/mL）,倒置显微镜下观察各组细胞的形态差异;采用Griess法和ELISA法测定各组细胞上清中一氧化氮（NO）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）的含量;采用qRT-PCR法检测各组细胞中TNF-α、IL-6和IL-1β的mRNA表达水平;利用免疫荧光法观察核因子-κBp65（p65）核移位情况;采用Westernblot法检测各组细胞一氧化氮合酶（iNOS）和p65蛋白表达情况。[结果] 0.01~10μg/mL清感冬饮对HEK293T细胞活力无显著影响,1~10μg/mL清感冬饮明显抑制TNF-α诱导的NF-κB启动子的活性,清感冬饮对ARE启动子的活性无显著影响。初步证明清感冬饮具有抗炎作用,而无明显抗氧化作用。0.01~100μg/mL清感冬饮干预24h对RAW264.7巨噬细胞无细胞毒性作用。与模型组相比,1~100μg/mL清感冬饮显著抑制LPS诱导的RAW264.7巨噬细胞中NO及炎症因子TNF-α、IL-6和IL-1β的释放;10~100μg/mL清感冬饮显著下调TNF-α、IL-6和IL-1β的mRNA表达水平,抑制p65核移位;100μg/mL清感冬饮显著抑制总蛋白iNOS、核蛋白p65表达,显著促进浆蛋白p65表达。[结论] 清感冬饮可抑制LPS诱导的RAW264.7巨噬细胞的炎症反应,该抗炎作用可能与调控NF-κB/iNOS/NO信号通路、抑制促炎因子释放有关。     

丹参酮ⅡA对TNF-α损伤平滑肌细胞MCP-1、IL-1β表达的影响

[目的]探讨丹参酮ⅡA对血管平滑肌细胞单核细胞趋化蛋白-1(MCP-1)、白细胞介素-1β(IL-1β)表达的影响。[方法]培养大鼠主动脉平滑肌细胞,复制肿瘤坏死因子-α(TNF-α)损伤模型,逆转录-聚合酶链反应(RT-PCR)、放射性免疫方法检测MCP-1、IL-1β的表达,并观察丹参酮ⅡA的作用。[结果]丹参酮ⅡA能够抑制TNF-α诱导的血管平滑肌细胞MCP-1、IL-1β的过度表达。[结论]抑制动脉粥样硬化过程中MCP-1、IL-1β的表达可能是丹参酮ⅡA的作用机制之一。     

针刺对慢性疲劳大鼠血清IL-2及TNF-α的影响

[目的]观察针刺对慢性疲劳大鼠血清白细胞介素-2(IL-2)及肿瘤坏死因子-α(TNF-α)的影响。[方法]采用慢性束缚和冷水游泳制备慢性疲劳大鼠模型。随机将大鼠分为正常对照组、模型组、针刺组和人参皂苷组。针刺组选百会、肾俞、后三里穴,针刺1次/d。人参皂苷组给予人参皂苷水溶液[70mg/(kg·d)]灌胃治疗,1次/d。正常对照组和模型组进行与针刺组和人参皂苷组相同时间、相同程度的捉抓。连续治疗2周断头取血。采用放射免疫分析法检测大鼠血清IL-2及TNF-α的水平。[结果]模型组大鼠血清IL-2、TNF-α水平均降低,针刺后两者均明显升高。[结论]针刺可以升高慢性疲劳大鼠血清IL-2、TNF-α水平,这可能是针刺治疗慢性疲劳综合征的作用机制之一。     

20-羟基蜕皮甾酮通过Akt/NF-κB通路调节小胶质细胞活化

目的 观察20-羟基蜕皮甾酮对脂多糖诱导的体外原代培养小胶质细胞活化的影响,并探讨其相关机制。方法 脂多糖处理原代培养的SD大鼠小胶质细胞,构建其活化模型。实验分为正常组,脂多糖处理组和脂多糖+20-羟基蜕皮甾酮组。酶联免疫法测定各组细胞培养基中IL-1β和TNF-α的浓度,Western blot法检测各组细胞胞质内p-IκBα、细胞核内p-NF-κB以及细胞内p-Akt水平。结果 10ng/ml LPS孵育小胶质细胞8h,细胞培养基中IL-1β和TNF-α浓度分别从33.73±6.42pg/ml和43.67±7.17pg/ml上升至87.16±12.78pg/ml和96.55±13.76pg/ml （P<0.01）。50μmol/L 20-羟基蜕皮甾酮和LPS一起处理细胞8h后,笔者观察到细胞上清中IL-1β和TNF-α浓度分别降低至59.37±9.24和72.81±12.69pg/ml （P<0.05）。进一步增加20-羟基蜕皮甾酮浓度至100μmol/L,细胞上清中IL-1β和TNF-α浓度进一步降低至48.11±8.42pg/ml和61.44±9.38pg/ml （P<0.01）。此外,脂多糖导致小胶质细胞胞质内p-IκBα、细胞核内p-NF-κB和细胞内p-Akt水平明显升高（P<0.01）,20-羟基蜕皮甾酮抑制上述蛋白水平的升高。结论 20-羟基蜕皮甾酮经Akt信号通路,失活NF-κB,抑制LPS诱导的小胶质细胞IL-1β和TNF-α分泌,最终改善小胶质细胞介导的炎症。     

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.