桃红四物汤促进自噬抑制氧糖剥夺诱导心肌细胞凋亡的实验研究

目的探讨桃红四物汤(TST)对氧糖剥夺(OGD)诱导H9C2心肌细胞损伤的保护作用。方法将H9C2心肌细胞分为对照组、OGD模型组、OGD+TST含药血清组、OGD+TST含药血清+雷帕霉素组、OGD+TST含药血清+氯喹组。采用四唑盐(MTT)比色法检测细胞活力,流式细胞术检测细胞凋亡,免疫印迹法(Western blot)检测微管相关蛋白轻链3Ⅱ/3Ⅰ(LC3-Ⅱ/LC3-Ⅰ)比值及核孔糖蛋白62(p62),B-细胞淋巴瘤因子2(Bcl-2)和B-细胞淋巴瘤因子2相关X蛋白(Bax)表达量。结果与对照组比较,OGD模型组细胞活力显著下降[(42.23±3.19)%比(100.00±1.54)%,P0.05],细胞凋亡比例显著增多[(32.14±2.88)%比(3.54±0.23)%,P0.05],p62和Bax蛋白表达增多,LC3-Ⅱ/LC3-Ⅰ比值和Bcl-2蛋白表达减少;与OGD模型组比较,OGD+TST含药血清组细胞活力显著升高[(80.04±5.19)%比(42.23±3.19)%,P0.05],细胞凋亡明显减少[(10.01±1.04)%比(32.14±2.88)%,P0.05],LC3-Ⅱ/Ⅰ比值、Bcl-2表达增多,p62和Bax蛋白表达减少。与OGD+TST含药血清组比较,OGD+TST含药血清+雷帕霉素组细胞活力显著升高[(89.01±5.35)%比(80.04±5.19)%,P0.05],细胞凋亡比例显著减少[(7.45±0.13)%比(10.01±1.04)%,P0.05],LC3-Ⅱ/Ⅰ比值、Bcl-2表达增多,p62和Bax蛋白表达减少;OGD+TST含药血清+氯喹组细胞活力下降[(53.13±4.13)%比(80.04±5.19)%,P0.05],凋亡细胞比例升高[(17.42±1.93)%比(10.01±1.04)%,P0.05],LC3-Ⅱ/Ⅰ比值、Bcl-2表达减少,p62和Bax蛋白表达增多。结论桃红四物汤可能促进氧糖剥夺H9C2心肌细胞自噬,抑制心肌细胞凋亡,对心肌缺血起保护作用。     

桃红四物汤促进自噬抑制氧糖剥夺诱导的心肌细胞凋亡的实验研究

目的 探讨桃红四物汤(TST)对氧糖剥夺(OGD)诱导的H9C2心肌细胞损伤的保护作用。 方法 将H9C2心肌细胞分为5组(n=3)：对照组、OGD模型组、OGD +TST含药血清处理组、OGD +TST含药血清+雷帕霉素处理组、OGD +TST含药血清+氯奎处理组。四唑盐(MTT)比色法检测细胞活力;流式细胞术检测细胞凋亡;免疫印迹法(western blot)检测自噬相关蛋白(LC3、p62)和凋亡相关蛋白(Bax、Bcl-2)的表达。结果 与对照组相比,OGD模型组细胞活力显著下降(P<0.05),细胞凋亡显著增多(P<0.05),LC3II/LC3I比值和促凋亡蛋白Bax显著增多,p62和抗凋亡蛋白Bcl-2表达减少。与OGD模型组相比,OGD +TST含药血清处理组的细胞活力显著升高(P<0.05),细胞凋亡明显减少(P<0.05),LC3II/LC3I比值、Bcl-2表达增多,p62和Bax蛋白表达减少。与OGD +TST含药血清处理组相比,OGD +TST含药血清+雷帕霉素处理组的细胞活力进一步升高(P<0.05),细胞凋亡比例进一步减少(P<0.05),LC3II/LC3I比值、Bcl-2表达进一步增多,p62和Bax蛋白表达进一步减少;而氯奎处理桃红四物汤治疗组的细胞活力下降(P<0.05),凋亡细胞比例升高(P<0.05),LC3II/LC3I比值、Bcl-2表达减少,p62和Bax蛋白表达增多。结论 桃红四物汤促进氧糖剥夺H9C2心肌细胞自噬,抑制心肌细胞凋亡,对心肌缺血起保护作用。     

miR-451a对慢性心衰细胞模型心肌细胞H9c2凋亡与自噬的影响

目的探究miR-451a对慢性心衰细胞模型心肌细胞H9c2凋亡与自噬的作用机制。 方法采用1 μmol/L阿霉素(DOX)建立慢性心衰细胞模型。将细胞实验分为Control组(正常心肌细胞H9c2)、DOX组(DOX培养)、DOX+miR-NC组(DOX培养转染对照miR-NC)、DOX+miR-451a组(DOX培养转染miR-451a模拟物miR-451a)、DOX+miR-451a+CCI-779组(DOX和CCI-779培养转染miR-451a)。qRT-PCR检测各组细胞miR-451a相对表达水平；CCK-8和流式细胞术分别检测细胞增殖活性与凋亡；Western blotting检测剪切的Caspase-3(cleaved Caspase-3)、Bcl-2相关X蛋白(Bax)、Beclin-1、微管相关蛋白1轻链3I(LC3I)、LC3II、哺乳动物雷帕霉素靶蛋白(mTOR)、p70s6k、磷酸化mTOR(p-mTOR)、p-p70s6k蛋白表达。 结果成功建立慢性心衰细胞模型。与Control组比较,DOX组miR-451a相对表达水平、细胞活性、p-mTOR、p-p70s6k蛋白表达降低,而凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达升高。与DOX+miR-NC组比较,DOX+miR-451a组miR-451a相对表达水平、细胞活性、p-mTOR、p-p70s6k蛋白表达升高,而凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达降低。与DOX+miR-451a组比较,DOX+miR-451a+CCI-779组细胞活性降低,凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达升高。 结论miR-451a通过抑制mTOR信号通路促进慢性心衰细胞模型心肌细胞H9c2凋亡和自噬。     

miR-425-5p通过靶向HSPB8表达介导心肌缺血再灌注损伤

目的：探讨miR-425-5p及其下游靶点HSPB8对心肌缺血再灌注损伤（MIRI）的影响及相关机制。方法：将AC16细胞随机分为control组,H/R组,H/R+miR-NC组,H/R+miR-425-5p inhibitor组,H/R+miR-425-5p mimic组,H/R+OE-NC组,H/R+OEHSPB8组,细胞计数试剂盒-8(CCK-8)法测定AC16的增殖能力;酶联免疫吸附测定（ELISA）法检测超氧化物歧化酶（SOD）、丙二醛（MDA）、肿瘤坏死因子-α(TNF-α)、白细胞介素（IL）-6水平,实时荧光定量PCR(RT-qPCR)和蛋白质免疫印迹法（western blotting）分别检测凋亡相关蛋白Caspase-3、Bax、Bcl-2的mRNA和蛋白表达情况,荧光素酶报告基因检测验证miR-425-5p及HSPB8间的靶向关系。结果：miR-425-5p在MIRI患者中表达显著高于正常人群;HSPB8在MIRI患者中表达低于正常人群;与control组相比,H/R组、H/R+miR-NC组、H/R+miR-425-5p mimic组、H/R+OE-NC组细...     

MiR-873对缺氧复氧诱导的心肌细胞凋亡的影响及其机制

目的：探讨微小RNA(micro RNA,miR)-873在心肌细胞缺氧复氧(hypoxia reoxygenation,H/R)损伤中的作 用及其相关机制。方法：采用体外培养大鼠H9c2心肌细胞构建H/R模型,并转染miR-873模拟物(mimic),实验分为对 照组、H/R组、阴性对照组及miR-873 mimic组。Real-time PCR检测各组miR-873的表达水平;蛋白质印迹法检测egl-9 家庭缺氧诱导因子3(egl-9 family hypoxia inducible factor 3,Egln3)、B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)和Bcl-2 相关X蛋白(Bcl-2 associated X protein,Bax)的表达水平;ELISA试剂盒以及天冬氨酸特异性半胱氨酸蛋白酶-3(cysteinecontaining, aspartate-specific proteases-3,caspase-3)活性检测试剂盒分别用来检测细胞凋亡和caspase-3活性;采用双荧 光素酶报告基因系统检测miR-873对Egln3的作用靶点,并将实验分为阴性对照组、携带野生型(wild type,WT)报告 基因的Egln3 3'-非编码区(3'-untranslated regions,3'-UTR)组(WT组)和携带突变型(mutant type,MUT)报告基因的Egln3 3'-UTR组(MUT组)。为进一步检测Egln3对miR-873 mimic作用的影响,实验构建了Egln3过表达细胞,并将其分为H/R 组、H/R+miR-873 mimic组、H/R+pcDNA3-Egln3(pcEgln3)组和H/R+miR-873 mimic+pcEgln3组。结果：与对照组相比, H/R组中miR-873表达显著下调,差异有统计学意义(P<0.05)。与H/R组相比,miR-873 mimic组中H9c2心肌细胞凋亡 和caspase-3活性下降,Bax/Bcl-2比值下降,差异均有统计学意义(均P<0.05);荧光素酶活性检测结果表明：与阴性对 照组相比,WT组荧光素酶活性显著下调,差异有统计学意义(P<0.05),而MUT组荧光素酶活性无明显变化,差异无 统计学意义(P>0.05)。过表达实验结果表明：与H/R组相比,miR-873 mimic组细胞凋亡和Bax/Bcl-2比值均显著下降(均 P<0.05);与miR-873 mimic组相比较,H/R+pcEgln3组和H/R+miR-873 mimic+pcEgln3组细胞凋亡和Bax/Bcl-2比值显著上 调,差异均有统计学意义(均P<0.05)。结论：MiR-873通过靶向作用于Egln3而抑制H/R心肌细胞凋亡。     

miR-1通过PI3K/Akt/mTOR信号通路对高糖诱导的H9c2心肌细胞自噬的影响

目的 探讨miR-1对高糖诱导的H9c2心肌细胞自噬及磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路的影响。方法H9c2细胞随机分为4组。正常组使用普通培养基培养,未转染;诱导组使用含33 mmol/L葡萄糖的培养基诱导,未转染;miR-1 NC组使用含33 mmol/L葡萄糖的培养基诱导,转染miR-1 NC;miR-1 inhibitor组使用含33 mmol/L葡萄糖的培养基诱导,转染miR-1 inhibitor。在加入葡萄糖诱导前48 h,使用Lipofectamine 3000转染试剂盒进行转染。实时荧光定量聚合酶链式反应（qRT-PCR）检测H9c2细胞和高糖诱导的H9c2细胞中miR-1表达水平;CCK-8法检测各组H9c2细胞增殖活性;蛋白免疫印记法（Western blot）检测各组H9c2细胞中Beclin1、p65、PI3K蛋白表达水平、LC3-Ⅱ/LC3-Ⅰ比值以及AKT、mTOR蛋白磷酸化水平;生物信息学预测和双萤光素酶验证miR-1和PI3K的靶向关系。结果与正常组相比,诱导组的H9c2细胞中,miR-1表达水平显著升高（P<0.05）,PI3K蛋白表达水平显著降低（P<0.05）。与正常组相比,诱导组H9c2细胞Beclin1蛋白表达、LC3-Ⅱ/LC3-Ⅰ比值显著升高,自噬小体增多,细胞增殖率、p62蛋白表达以及PI3K、Akt、mTOR蛋白磷酸化水平显著降低（P<0.05）;与诱导组和miR-1 NC组相比,miR-1 inhibitor组H9c2细胞miR-1水平、Beclin1蛋白表达、LC3-Ⅱ/LC3-Ⅰ比值显著降低,自噬小体减少,细胞增殖率、p62蛋白表达以及PI3K、Akt、mTOR蛋白磷酸化水平显著升高（P<0.05）;miR-1和PI3K间存在靶向关系。结论在高糖诱导的H9c2细胞中,miR-1表达水平升高,抑制miR-1,可通过靶向激活PI3K/Akt/mTOR信号通路,抑制高糖诱导的H9c2心肌细胞自噬。     

参芪益心方抑制阿霉素诱导心肌细胞凋亡作用研究

目的:证实参芪益心方可以抑制阿霉素诱导的心肌细胞凋亡。方法:用SD大鼠制备含药血清,用阿霉素诱导H9c2心肌细胞凋亡建立模型,将细胞设置为空白对照组、模型组、含药血清高剂量组(8%)、含药血清中剂量组(4%)、含药血清低剂量组(2%)和卡托普利组。含药血清提前预处理,阿霉素造模72 h后进行试验检测,倒置显微镜下观察H9c2心肌细胞形态,MTT法检测细胞增殖活性;Annexin V-FITC/PI双染法检测H9c2心肌细胞凋亡。结果:H9c2心肌细胞形态:参芪益心方含药血清高、中、低剂量组及卡托普利组可改善细胞凋亡现象,其中以含药血清高剂量组、卡托普利组改善最为明显。MTT法检测H9c2心肌细胞增殖率:模型组OD值、增殖率与空白对照组比较差异具有统计学意义(P0.01);含药血清高、中、低剂量组及卡托普利组OD值、增殖率与模型组比较差异具有统计学意义(P0.05,P0.01);含药血清高剂量组与卡托普利组OD值比较,差异无统计学意义(P0.05)。流式细胞仪测得结果示:与空白对照组比较,模型组细胞凋亡数量(Q2+Q4)明显增多;含药血清预处理后(Q2+Q4)明显降低,含药血清高、中剂量组及卡托普利组与模型组比较差异具有统计学意义(P0.01)。结论:参芪益心方可以抑制阿霉素诱导的细胞凋亡。     

丙烯醛对缺氧复氧H9c2心肌细胞损伤的影响及机制初探

目的研究丙烯醛对缺氧复氧H9c2心肌细胞损伤的影响,并探讨其促凋亡作用机理。方法建立H9c2心肌细胞缺氧复氧(H/R)损伤模型,分为正常对照组、丙烯醛组(ACR)、缺氧复氧组(H/R)、丙烯醛+缺氧复氧组(ACR+H/R)。用10μmol/L丙烯醛预处理H9c2心肌细胞30min后2h缺氧16h复氧,采用MTT法检测细胞存活率,4′,6-二脒基-2-苯基吲哚(DAPI)荧光染色检测细胞核凋亡的形态学改变,流式细胞术检测细胞凋亡,Western blot检测凋亡相关蛋白cytochrome c(cyt c)、caspase 9和caspase 3的表达水平。结果与H/R组相比,ACR+H/R组H9c2细胞存活率下降、凋亡增加(P<0.05),cyt c的线粒体蛋白表达水平下调、细胞浆蛋白表达水平上调,caspase 9和caspase 3蛋白表达水平下调并出现明显裂解蛋白。结论作为一种来源于人类生活环境的心血管毒物,丙烯醛能加剧缺氧复氧H9c2心肌细胞的损伤,可能与cyt c由线粒体释放到细胞浆、激活caspase级联反应导致线粒体功能损伤有关。     

SSTF上调miR-495表达减缓高糖诱导的视网膜神经节细胞氧化损伤和凋亡

目的 探讨黄岑茎叶总黄酮(SSTF)对高糖诱导的视网膜神经节细胞(RGC-5)损伤的保护机制。方法 根据处理方法不同将RGC-5细胞分成高糖(HG)组、对照(Con)组、HG+SSTF-低剂量(L)组、HG+SSTF-中剂量(M)组、HG+SSTF-高剂量(H)组、HG+miR-NC组、HG+miR-495组、HG+SSTF+anti-miR-NC组、HG+SSTF+anti-miR-495组，每组设置9复孔。检测MDA含量以及CAT、SOD活性。流式细胞术检测细胞凋亡。实时荧光定量PCR(qRT-PCR)检测miR-495表达。结果 HG组RGC-5细胞凋亡率、MDA含量高于Con组(P<0.05),CAT、SOD的活性、miR-495表达低于Con组(P<0.05)。HG+SSTF-L组、HG+SSTF-M组、HG+SSTF-H组RGC-5细胞凋亡率、MDA含量低于HG组(P<0.05),CAT和SOD的活性、miR-495表达高于HG组(P<0.05)。HG+miR-495组RGC-5细胞凋亡率[(14.96±1.26)%]、MDA含量[(4.62±0.4...     

熊果酸通过抑制自噬对血清剥夺诱导的H9c2细胞起抗凋亡作用

目的冠状动脉粥样硬化心脏病(coronary atherosclerotic cardiopathy)造成的心肌缺血(myocardial ischemia)严重威胁人类生命安全,本研究从细胞水平阐述了熊果酸(ursolic acid,UA)对血清剥夺心肌细胞自噬和凋亡的作用。方法体外培养大鼠H9c2心肌细胞。分为对照组、血清剥夺组、熊果酸处理组、自噬抑制剂3-甲基腺嘌呤(3-MA)预处理联合熊果酸组、雷帕霉素预处理联合熊果酸组。流式细胞术检测细胞凋亡情况;蛋白免疫印迹检测自噬相关蛋白LC3、Beclin1、ATG5以及凋亡相关蛋白cleaved-caspase3、Bc L-2、Bax的表达。结果10μmol/L熊果酸为处理H9c2细胞的最适浓度;血清剥夺能造成H9c2细胞LC3 II/I比值、Beclin1和ATG5表达增加(P0. 01),熊果酸处理后能降低其表达(P0. 05);血清剥夺造成H9c2细胞凋亡率增加(P0. 001),熊果酸处理后能降低凋亡率(P0. 01),3-MA预处理组能进一步降低凋亡率(P0. 001),而雷帕霉素预处理组凋亡率上升(P0. 001);血清剥夺能造成H9c2细胞cleaved-caspase3表达量、Bax/Bc L-2比值升高(P0. 001),熊果酸处理后抑制了血清剥夺造成的cleaved-caspase3的表达量和Bax/Bc L-2比值升高(P0. 01),3-MA预处理组能进一步降低cleaved-caspase3的表达量和Bax/Bc L-2比值(P0. 001),雷帕霉素预处理组Bax/Bc L-2比值回升(P0. 05)。结论熊果酸对血清剥夺H9c2细胞的抗凋亡作用与抑制自噬有关。     

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…