消栓通络有效成分组对SH-SY5Y细胞氧糖剥夺损伤的影响

目的研究消栓通络有效成分组(XECG)对SH-SY5Y细胞氧糖剥夺(OGD)损伤的保护作用,并探讨其可能的作用机制。方法体外培养SH-SY5Y细胞,将细胞随机分为正常组、氧糖剥夺模型组和XECG组(1、3、10mg·L-1),建立体外OGD/R细胞模型。倒置显微镜观察细胞形态;MTT法测定细胞存活率;酶标法试剂盒测定乳酸脱氢酶(LDH)漏出量;Western Blot检测凋亡相关蛋白Bcl-2、Bax蛋白表达的变化。结果与模型组相比,XECG能明显改善OGD/R引起的SH-SY5Y细胞的损伤,提高细胞存活率(P<0.05),减少LDH的释放量(P<0.05),能提高缺糖缺氧神经元Bcl-2/Bax比值。结论 XECG对OGD/R引起的SH-SY5Y细胞损伤有保护作用,其机制可能与影响凋亡相关基因Bcl-2、Bax的表达有关。     

实验性颅脑损伤鼠垂体前叶HIF-1α、Bcl-2/Bax的表达和细胞凋亡的研究

目的：探讨颅脑损伤后垂体前叶细胞缺氧诱导因子-1α（HIF-1α）、Bcl-2和Bax的表达及细胞凋亡情况。方法：采用Feeney自由落体方法制作大鼠颅脑损伤模型,分别于建模后24、72 h和7 d灌注固定取材,同时设立对照组,免疫组化染色法检测HIF-1α、Bcl-2和Bax蛋白的表达,TUNEL染色法检测细胞凋亡。结果：垂体前叶组织于伤后24 h HIF-1α、Bcl-2和Bax蛋白的表达即有升高,72 h达到高峰,一直持续到伤后7 d,与对照组相比较差异均有统计学意义（P〈0.05）;TUNEL染色显示凋亡细胞明显增多,且凋亡指数与HIF-1α的表达和Bcl-2/Bax的比值成规律性变化。结论：颅脑损伤后垂体前叶组织HIF-1α、Bc1-2、Bax蛋白的表达增加而诱发细胞凋亡,是颅脑损伤后垂体前叶功能紊乱的重要原因。     

乌司他丁对缺血性损伤兔海马HIF-1α表达的影响

目的 探讨乌司他丁(UTI)对控制性降压(CH)诱导的缺血缺氧性损伤的兔海马CA1神经元缺氧诱导因子-1α(HIF-1α)表达的影响.方法 36只健康雄性新西兰白兔随机分成假手术对照组(Cont组)、控制性降压组(CH组)和UTI预处理组(UTI+CH组),每组12只.构建硝普钠CH模型,CH组和UTI+CH组平均动脉压(MAP)均降至基础值的50%,UTI+CH组在降压前30 min静脉注射UTI 50000 U/kg.Western blot检测HIF-1α、Bcl-2和Bax蛋白的表达水平,TUNEL染色观察海马CA1细胞凋亡,HIF-1α免疫组化染色观察CA1神经元损伤状况.结果 UTI+CH组HIF-1α和Bax蛋白表达水平明显低于CH组(P<0.05),而其Bcl-2的表达则明显比CH组高(P<0.05).TUNEL染色的细胞凋亡指数CH组明显高于Cont组(P<0.05),而UTI+CH组则比CH组低(P<0.05).结论 UTI可能通过抑制HIF-1α蛋白和促凋亡蛋白Bax的表达,上调抗凋亡蛋白Bcl-2的表达,从而减轻CH诱导的兔海马缺血缺氧性损伤.     

IGF-1R/PI3K信号通路介导胰岛素样生长因子-1抗MPP~+诱导多巴胺能神经元损伤的作用机制研究

目的观察不同浓度的胰岛素样生长因子-1 (IGF-1)对1-甲基-4-苯基吡啶离子(MPP~+)诱导的多巴胺能神经元细胞系SH-SY5Y细胞损伤的保护作用、胰岛素样生长因子-1受体(IGF-1R)阻断剂和磷脂酰肌醇-3-激酶(PI3K)阻断剂LY294002的阻断效应。方法体外培养人神经母细胞瘤SH-SY5Y细胞,应用不同浓度的IGF-1 (25、50及100 ng/mL)预处理细胞24 h,然后与MPP~+(1 mM)共同处理细胞24 h;提前1h加入JB-1 (1μg/mL)或LY294002 (5μM),然后加入IGF-1预保护细胞24 h,与MPP~+共同处理细胞24 h。应用Western blotting检测凋亡因子Bax和Bcl-2的蛋白变化,观察IGF-1的抗凋亡作用及阻断剂对IGF-1神经保护作用的阻断效应。结果 MPP~+处理组与对照组比较,Bax蛋白表达升高,而Bcl-2蛋白表达降低(P<0.05)。不同浓度的IGF-1与MPP~+处理组比较,Bax蛋白表达降低,而Bcl-2蛋白表达升高(P<0.05)。JB-1或LY294002能够阻断IGF-1的该作用(P<0.05)。结论不同浓度的IGF-1能够对抗MPP~+诱导的SH-SY5Y细胞凋亡,该作用与IGF-1R/PI3K信号通路有关。     

藁本内酯预处理对Aβ25-35诱导人脑神经瘤母细胞SH-SY5Y细胞毒性的保护作用

目的探讨藁本内酯(Z-ligustilide,LIG)对β淀粉样蛋白(Aβ)25-35引起的人脑神经瘤母细胞SH-SY5Y细胞毒性的保护作用。方法 0.1、1.0、2.5、5.0μg/mL LIG作用于SH-SY5Y细胞,24h后以50μmol/LAβ25-35诱导SH-SY5Y细胞凋亡建立阿尔茨海默病体外模型,MTT法检测细胞活性,Western blot法检测细胞中凋亡蛋白含量。结果经Aβ25-35处理后,细胞存活率下降,细胞内凋亡相关蛋白——Bax、cleaved caspase3、细胞色素C、caspase8表达上调,但Bcl-2蛋白表达下降(P均<0.05)。细胞经过LIG一定浓度预处理,细胞存活率提高,凋亡相关蛋白——Bax、cleaved caspase3、细胞色素C、caspase8蛋白表达下降,Bcl-2表达上升(P均<0.05)。结论 LIG具有对Aβ25-35诱导的SH-SY5Y细胞毒性的保护作用,其机制可能是抑制Aβ诱导的细胞凋亡。     

氯化钴预处理在培养分化的人神经母细胞瘤细胞株缺氧损伤中的保护作用

目的 探讨氧化钴(CoCl2)对体外培养分化的人神经母细胞瘤SH-SY5Y细胞株的缺氧损伤保护作用及缺氧诱导因子1、2和其调节基因产物血管内皮生长因子在其中的作用.方法 分化的SH-SY5Y细胞分为对照组、化学缺氧预处理组(细胞用50 μmol/L CoCl2预处理3 h,换液后常氧培养1 h,然后在2%的低氧孵箱内缺氧28 h)、缺氧组(无CoCl2预处理过程,其余同化学缺氧预处理组).用Western blotting法测细胞HIF-1α、HIF-2α和VEGF的蛋白表达,通过乳酸脱氢酶释放率测定、MTT细胞活力测定判断细胞损伤程度.结果 化学缺氧预处理组细胞 HIF-2α和VEGF的蛋白表达显著高于缺氧组(P＜0.01),HIF-1α蛋白表达在两组之间没有显著性差异(P＞0.05),化学缺氧预处理组细胞较缺氧组细胞存活率高、乳酸脱氢酶释放率减少(P＜0.01).结论 CoCl2化学预缺氧可保护分化的SH-SY5Y细胞对缺氧产生耐受,缺氧预处理可能通过HIF-2促进VEGF的表达而产生保护作用.     

表没食子儿茶素没食子酸酯对稳定表达淀粉样前体蛋白的SH-SY5Y细胞抗炎抗凋亡作用研究

目的 探讨表没食子儿茶素没食子酸酯(epigallocatechingallate,EGCG)对稳定表达淀粉样前体蛋白(amyloid precursor protein, APP)的SH-SY5Y细胞抗炎抗凋亡的保护作用.方法 将SH-SY5Y细胞分为对照组、模型组(稳定表达APP的细胞)和EGCG组(APP+10、20、30 μmol/L的EGCG).免疫荧光细胞化学法观察APP在各组细胞中表达;MTT法检测细胞存活能力;TUNEL法检测细胞凋亡情况;RT-PCR法检测Bax、Bcl-2和Caspase-3表达;ELISA法测定IL-6和TNF-α浓度.结果 模型组中,APP高表达;与模型组比较,给予20 μmol/L EGCG的EGCG组明显提高SH-SY5Y细胞的存活率,减少细胞凋亡数目,下调Bax/Bcl-2比例及Caspase-3 mRNA表达,减少IL-6和TNF-α浓度.结论 EGCG对稳定表达APP的SH-SY5Y细胞有一定保护作用,可能是通过抗炎以及减少细胞凋亡发挥治疗作用.     

慢病毒介导HIF-1α沉默对缺氧肝癌细胞HepG2增殖和凋亡的影响

目的:构建缺氧诱导因子-1α(HIF-1α)-短发夹RNA(shRNA)慢病毒表达载体,观察慢病毒介导的HIF-1α基因沉默对体外模拟缺氧条件下的肝癌细胞HepG2增殖和凋亡的影响及其机制。方法:设计合成1条针对HIF-1α基因的shRNA干扰序列,构建于慢病毒载体质粒GV248中,转染293T细胞包装产生慢病毒,转染人肝癌细胞株HepG2细胞,嘌呤霉素筛选出稳定转染的细胞株。将HepG2细胞分为正常对照组、干扰对照组(转染NC-shRNA)、干扰组(转染HIF-1α-shRNA),应用化学缺氧法(150μmol/L CoCl2)模拟肿瘤细胞缺氧微环境,应用Western Blot检测HepG2细胞中HIF-1α蛋白表达证实基因沉默效果,CCK-8检测沉默HIF-1α基因对HepG2细胞增殖的影响,Annexin V-FITC凋亡检测试剂盒和流式细胞仪检测沉默HIF-1α基因对细胞凋亡的影响。Western Blot检测HepG2细胞内凋亡相关蛋白Bcl-2、Bax、Cleaved caspase-3、Cleaved caspase-9和细胞质细胞色素C(Cyt-C)表达的变化。结果:①成功构建HIF1α-shRNA慢病毒载体并转染HepG2细胞,缺氧培养细胞24h后,与正常对照组及干扰对照组相比,干扰组细胞内HIF-1α蛋白表达明显降低(P<0.01),证实慢病毒载体能够有效沉默HIF-1α基因。②沉默HIF-1α基因后缺氧培养的HepG2细胞增殖率下降(P<0.05),而细胞凋亡比例升高(P<0.01)。③Western Blot结果显示,与正常对照组及干扰对照组相比,沉默HIF-1α基因后细胞内凋亡相关蛋白Bax、Cleaved caspase-3、Cleaved caspase-9和细胞质Cyt-C表达上调(P<0.05或P<0.01),而抗凋亡蛋白Bcl-2表达下调(P<0.01),Bax/Bcl-2蛋白表达比例升高。结论:通过shRNA慢病毒载体途径可有效沉默缺氧HepG2肝癌细胞HIF-1α基因,HIF-1α沉默能显著抑制缺氧HepG2细胞增殖活性,促进细胞凋亡,其凋亡途径可能与激活内源性线粒体凋亡通路有关。     

瘦素减轻鱼藤酮诱导的SH-SY5Y细胞损伤

目的 探讨瘦素对鱼藤酮（rotenone） 所诱导的人神经母细胞瘤SH-SY5Y 细胞凋亡的抑制作用。 方法 建立鱼藤酮诱导的SH-SY5Y 细胞损伤模型,用瘦素进行干预,采用MTT 比色法检测细胞存活率,锥虫蓝检测细胞死亡率,细胞形态学观察,并用Western blot 法检测Bcl-2、Bax、p-GSK-3β（Ser9） 和GSK-3β 的表达水平,免疫荧光检测α-synuclein 的表达水平。 结果 成功建立了鱼藤酮诱导的SH-SY5Y 细胞损伤模型。在模型的基础上,瘦素干预后,细胞状态改善,细胞存活率提高约14%（P ＜ 0.05）,细胞死亡率降低约17%（P ＜ 0.05）,α-synuclein 的浓度下降,Bcl-2 和p-GSK-3β 的表达显著提高。 结论 瘦素对鱼藤酮诱导的SH-SY5Y 细胞损伤具有明显的抑制作用,可能是通过抑制GSK-3β 的活性上调Bcl-2/Bax 的比率和降低α-synuclein 的表达而得以实现。     

丙烯酰胺对SH-SY5Y神经细胞凋亡和 miR-21表达的影响

目的:探讨丙烯酰胺(AA)对人SH-SY5Y神经细胞凋亡和miR-21表达的影响?方法:不同浓度丙烯酰胺作用于人神经母细胞瘤SH-SY5Y细胞24 h后,采用四甲基偶氮唑盐(MTT)法测定丙烯酰胺对SH-SY5Y细胞活力的影响,Hoechst 33258荧光染色法检测细胞凋亡,real-time PCR检测miR-21表达水平,Western blot检测PTEN?p-AKT?AKT?Bcl-2?Bax?caspase 9 和caspase 3蛋白表达?结果:丙烯酰胺剂量依赖性地降低SH-SY5Y细胞活性,Hoechst 33258染色显示明显细胞凋亡形态变化;丙烯酰胺显著降低SH-SY5Y细胞miR-21的表达,同时升高PTEN?Bax?caspase 9?caspase 3水平并降低p-AKT和Bcl-2表达?结论:丙烯酰胺可通过线粒体途径诱导SH-SY5Y神经细胞凋亡,其机制可能与丙烯酰胺引起miR-21表达下调有关?     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.