槲皮素抑制ADMA引起的脐静脉内皮细胞内质网应激和细胞凋亡

目的 探讨槲皮素改善ADMA引起脐静脉内皮细胞凋亡的可能机制。方法 体外培养人脐静脉内皮细胞,应用流式细胞仪技术检测细胞凋亡情况,应用Western blot法检测内质网应激蛋白PERK磷酸化水平及IRE1的蛋白表达量,用RT-PCR检测ATF4及CHOP mRNA表达量。结果 ADMA诱导脐静脉内皮细胞凋亡,槲皮素可以抑制ADMA引起的PERK磷酸化及IRE1蛋白表达,并可以减少ATF4及CHOP mRNA量,继而改善ADMA引起的细胞凋亡。结论 槲皮素抑制ADMA引起的脐静脉内皮细胞内质网应激继而抑制细胞凋亡。     

马兜铃酸对人脐静脉血管内皮细胞凋亡的影响

目的 研究马兜铃酸(AA)对人脐静脉血管内皮细胞(HUVECs)凋亡的影响.方法 通过Hoechst33258荧光染色观察细胞核形态改变,流式细胞仪检测细胞凋亡率,western blot法检测凋亡蛋白bcl-2和bax的表达及比色法测定caspase-3的活性.结果 马兜铃酸可引起内皮细胞核出现凋亡形态学改变,且呈浓度依赖方式增加了内皮细胞凋亡率.同时可降低内皮细胞内bcl-2蛋白的表达,增加bax蛋白的表达,使caspase-3活性增加.结论 马兜铃酸可抑制内皮细胞内bcl-2蛋白的表达,促进bax的表达,增加caspase-3的活性,从而使内皮细胞出现凋亡.     

阿托伐他汀对大鼠脑缺血再灌注PERK/elfR2a通路及Caspase-3表达的影响

目的 研究蛋白激酶R样内质网激酶(PERK)/eIFR2a通路及Caspase-3在大鼠脑缺血再灌注损伤中的作用机制及阿托伐他汀对其的影响。方法采用大脑中动脉线栓塞法制作大鼠脑缺血再灌注模型;随机分为缺血再灌注组、假手术组、阿托伐他汀组、阿托伐他汀+Salubrinal抑制剂组,大体标本采用TTC染色,釆用Western-blot法检测PERK、Caspase-3蛋白表达及eIF2a蛋白磷酸化。结果与假手术组相比,大鼠缺血再灌注后PERK蛋白表达及eIF2a 的磷酸化增加, Caspase-3表达的活性增强(P<0.01);阿托伐他汀干预可以减轻PERK蛋白表达及eIF2a蛋白磷酸化(P<0.05)。给予特异性eIF2a磷酸化抑制剂Salubrinal后可抑制eIF2a的磷酸化及Caspase-3表达的活性(P<0.05),对PERK蛋白表达无影响。形态学上从TTC染色提示:在缺血再灌注组TTC染色可见大片脑梗死组织。Salubrinal抑制剂及阿托伐他汀干预后脑梗死体积明显缩小(P<0.05)。结论内质网应激通过PERK/eIF2a/Caspase-3途径促进细胞凋亡,阿托伐他汀干预可以减轻脑缺血再灌注损伤。     

阿托伐他汀对同型半胱氨酸诱导内皮细胞内质网应激的抑制作用

目的： 观察阿托伐他汀对同型半胱氨酸（Hcy）诱导的人脐静脉内皮细胞内质网应激（endoplasmic reticulum stress,ERS）以及凋亡的影响。方法: 不同浓度的Hcy或联合阿托伐他汀处理人脐静脉内皮细胞后,流式细胞术检测细胞凋亡率,实时定量PCR和蛋白质印迹法测定ERS相关分子BiP、CHOP及p-eIF2α、p- PERK的表达变化。结果: Hcy呈浓度依赖性地诱导内皮细胞ERS相关分子BiP、CHOP及p-eIF2α、p-PERK的表达,阿托伐他汀可明显抑制Hcy诱导的内皮细胞ERS相关分子的表达,减少内皮细胞凋亡。结论： 阿托伐他汀可通过下调ERS相关信号通路,抑制Hcy诱导的人脐静脉内皮细胞凋亡。     

丹参酮ⅡA对马兜铃酸诱导的血管内皮细胞凋亡的保护作用及其可能机制

目的 探讨丹参酮ⅡA(TSN)对马兜铃酸(AA)诱导的血管内皮细胞凋亡的保护作用及其可能机制.方法 不同浓度的TSN(0.2、0.4、0.8 mg/L)与人脐静脉血管内皮细胞(HUVECs)培养1 h 后,加入10 mg/L的AA共同培养24 h.Hoechst 33258荧光染色观察细胞核的形态学改变,Annexin V-FITC/PI双染流式细胞仪检测细胞凋亡率,western blot法检测凋亡蛋白bcl-2和bax的表达及比色法测定caspase-3活性.结果 与正常对照组相比,AA组细胞凋亡率显著增加,同时抗凋亡蛋白bcl-2的表达降低,促凋亡蛋白bax的表达及caspase-3活性均升高;而经TSN干预后,能逆转AA对细胞凋亡率、凋亡蛋白bcl-2和bax的表达及caspase-3活性的上述作用.结论 TSN可抑制AA诱导的血管内皮细胞凋亡,这可能与其促进抗凋亡蛋白bcl-2的表达、抑制促凋亡蛋白bax的表达和下调caspase-3活性有关.     

神经酰胺对人内皮细胞凋亡及bax、bcl-2表达的影响

目的 观察外源性神经酰胺(C2-ceramide)对内皮细胞凋亡及相关基因bax和bcl-2表达变化的影响.方法 原代培养人脐静脉血管内皮细胞,加药组加入终浓度分别为5、12.5、25、50 μmol/L的C2-ceramide,对照组加入体积分数为0.1%的无水乙醇.采用TUNEL法检测细胞凋亡,用RT-PCR、Western blot技术对bax mRNA、bcl-2 mRNA及蛋白表达进行分析.结果 加药组内皮细胞的凋亡阳性率显著高于对照组(P＜0.05),且具有时间和剂量依赖性;加药组bax mRNA及蛋白的表达较对照组显著升高(P＜0.05),bcl-2 mRNA及蛋白的表达较对照组则显著降低(P＜0.05).结论 外源性神经酰胺通过上调bax及下调bcl-2表达水平,改变bax/bcl-2值,从而诱导内皮细胞凋亡.     

内质网应激参与高磷诱导的血管平滑肌细胞钙化的作用机制

目的 探讨内质网应激是否参与高磷诱导血管平滑肌细胞钙化及其作用机制。方法 体外培养人胸主动脉血管平滑肌细胞（VSMC）,分为对照组、正常磷组（1.3 mmol/L磷,NP组）、高磷组（2.6 mmol/L磷,HP组）,培养10 d。茜素红染色后观察细胞钙盐沉积情况;实时定量PCR和Western blotting检测肌醇需求因子1 (IRE1)、RNA依赖的蛋白激酶样内质网激酶（PERK）、内质网应激标志蛋白C/EBP同源蛋白（CHOP）、c-Jun氨基末端激酶（JNK）、骨形态发生蛋白2 (BMP-2)及Runt相关转录因子2 (Runx-2)的表达;应用CHOP siRNA转染高磷培养VSMC敲低CHOP表达后,检测BMP-2及Runx-2表达;应用JNK抑制剂SP600125 (DMSO溶解),终浓度10μmol/L预处理30 min后加入高磷培养基培养10 min,茜素红染色观察钙沉积情况。结果 与对照组及NP组比较,HP组内质网应激蛋白（IRE1、PERK、JNK、CHOP）及钙化蛋白（BMP-2、Runx-2）均明显增高（均P <0.01）。敲低CHOP表达后钙化蛋白...     

血管紧张素Ⅱ调控bax、bcl-2 mRNA表达诱导血管内皮细胞凋亡

目的研究血管紧张素Ⅱ（AngⅡ）诱导细胞凋亡的机制。方法健康胎儿脐静脉内皮细胞培养至第3代，用不同浓度AngⅡ作用18h，用TUNEL和ELISA检测AngⅡ能否诱导细胞凋亡及其量效关系；用RT—PCR检测bax和bcl-2的mRNA表达。结果．AngⅡ与2型血管紧张素Ⅱ受体（AT2）激动剂CGP42112A一样可以诱导血管内皮细胞凋亡；并且凋亡强度与AngⅡ是典型的剂量依赖关系。在AngⅡ与CGP4211A作用18h后，bax mRNA显著增加，bcl-2 mRNA显著减少。结论AngⅡ通过在转录水平调节bax和bcl-2 mRNA的表达，从而诱导血管内皮细胞凋亡。     

老年大鼠肾脏损伤中ASPP2蛋白调控与内质网应激的作用及相关性分析

目的 探讨P53凋亡刺激蛋白(ASPP)家族成员ASPP2蛋白及内质网应激在老年大鼠肾脏损伤中的作用机制.方法 以6月龄SD大鼠为成年组,18月龄SD大鼠为老年组,苏木精-伊红染色(HE)观察各组大鼠肾脏结构的改变;qRT-PCR检测内质网应激相关指标C/EPB同源蛋白(CHOP)、活化转录因子6(ATF6)、X盒结合蛋白-1(XBP-1)、葡萄糖调节蛋白78(GRP78)的mRNA表达水平;Western blot法检测各组大鼠ASPP2蛋白的表达水平,并对ASPP2的表达水平与内质网应激相关指标CHOP、ATF6、XBP-1、GRP78的表达水平作相关性分析.结果 与成年组SD大鼠相比,老年组SD大鼠肾脏结构明显改变,CHOP、ATF6、XBP-1、GRP78的mRNA表达水平及ASPP2蛋白表达水平显著增高,且ASPP2水平与CHOP和XBP-1呈正相关,ASPP2水平与ATF6呈负相关.结论 在老年SD大鼠中,ASPP2调控内质网应激可能是引起肾脏损伤的重要机制之一.     

高糖对胰岛β细胞增殖活力、分泌功能及内质网应激相关分子表达的影响

目的 研究持续高糖刺激对胰岛β细胞株MIN6增殖活力、分泌功能及内质网应激相关分子表达的影响.方法 将不同浓度葡萄糖(5.6、25和33.3mmol/L组)分别加入胰岛β细胞株MIN6培养液中,于不同时间点(24h和96h)收集细胞进行相关检测.CCK-8法检测细胞增殖活力;ELISA法检测胰岛素和胰岛素原分泌能力;Real-time PCR和Western blotting分别检测内质网应激相关分子Total XBP-1、Spliced XBP-1 mRNA及磷酸化IRE1α蛋白的表达.结果 加入葡萄糖后96h,各组细胞增殖活力均显著低于孵育后24h;在同一时间点,33.3mmol/L组和25mmol/L组显著低于5.6 mmol/L组(均P<0.05).加入葡萄糖后96h,33.3mmol/L组胰岛素分泌较干预后24h明显减少,而胰岛素原分泌显著增加(均P<0.05).加入葡萄糖后24h和96 h的各组Spliced XBP-1 mRNA表达,以及加入葡萄糖后96h的各组Total XBP-1 mRNA和磷酸化 IRE1α蛋白表达,均随葡萄糖浓度的增加而上调(P<0.05),呈现浓度依赖性.结论 持续性高糖可使胰岛β细胞的增殖活力下降、分泌功能受损及内质网应激相关分子表达增加,而后者可能是导致葡萄糖毒性的原因之一.     

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…     

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.     

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.     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…