虫草素对人骨肉瘤MG-63细胞凋亡的影响及其机制研究

目的 研究虫草素对人骨肉瘤MG-63细胞凋亡的影响及其机制。方法 采用CCK-8实验检测不同浓度虫草素作用于MG-63骨肉瘤细胞24h和48h后对癌细胞增殖的抑制作用;采用流式细胞术分析虫草素对MG-63细胞周期分布及凋亡的影响;采用Western blot法检测虫草素对MG-63细胞凋亡相关蛋白Bax、Bcl-2、cleaved caspase-9、cleaved caspase-3及NICD1、Hes1蛋白表达的影响。结果 虫草素对人骨肉瘤MG-63细胞有明显的增殖抑制作用,导致MG-63细胞周期阻滞于G₀/G₁期并诱导细胞凋亡;虫草素可上调Bax、cleaved caspase-9和cleaved caspase-3蛋白的表达并下调Bcl-2,NICD1和Hes1蛋白的表达。结论 虫草素通过抑制骨肉瘤细胞增殖,诱导细胞周期阻滞于G₀/G₁期并通过线粒体凋亡途径诱导MG-63细胞凋亡从而发挥抗骨肉瘤作用,可能骨肉瘤细胞中Notch信号通路活性下调有关。     

2-甲基-3-羟基蒽醌诱导人乳腺癌MCF-7细胞凋亡的机制

为探讨2-甲基-3-羟基蒽醌抗肿瘤作用及其机制,本研究采用锥虫蓝法检测细胞活力,流式细胞仪检测细胞周期变化、细胞凋亡率、线粒体膜电位及细胞内游离钙的变化,Western blot方法检测凋亡相关蛋白caspase-4、caspase-7、caspase-9、Bcl-2、Bax、JNK、细胞色素C的表达。结果发现：2-甲基-3-羟基蒽醌时间依赖性地抑制乳腺癌细胞的生长,升高细胞内游离钙含量,降低线粒体膜电位并诱导其凋亡;药物上调Bax并下调Bcl-2蛋白的表达;诱导caspase-4、caspase-7、caspase-9、calpain的活化及细胞色素C的释放。结果提示2-甲基-3-羟基蒽醌可能通过Ca²⁺/calpain/caspase-4途径诱导人乳腺癌MCF-7细胞凋亡。     

紫檀芪通过线粒体途径调控结直肠癌细胞增殖和凋亡

目的 探讨紫檀芪对结直肠癌细胞增殖和凋亡的作用及其机制。方法 使用梯度浓度的紫檀芪体外处理结直肠癌细胞,CCK-8法检测结直肠癌细胞增殖和活性,Hoechst 33258荧光染色法检测细胞凋亡,DCFH-DA荧光探针检测细胞内活性氧(reactive oxygen species, ROS)水平的变化,JC-1荧光探针检测细胞内线粒体膜电位的变化,Western blot法检测线粒体途径相关蛋白表达水平。结果 与对照组比较,紫檀芪对结直肠癌细胞增殖有抑制作用,且呈剂量和时间依赖性(P<0.05);Hoechst 33258染色和流式结果显示,紫檀芪促进结直肠癌细胞凋亡(P<0.05);紫檀芪诱导细胞内ROS产生、降低线粒体膜电位水平(P<0.05),增加Apaf-1、AIF、Bax、Cyt C、cleaved caspase-3和cleaved caspase-9的表达,降低Bcl-2、PCNA和survivin的蛋白水平(P<0.05)。结论 紫檀芪增加结直肠癌细胞内ROS水平、降低线粒体膜电位水平,通过线粒体途径抑制结直肠癌细胞增殖,诱导结直肠癌细胞凋亡。     

苦参碱通过线粒体通路诱导HepG2细胞凋亡的机制研究

目的探讨苦参碱通过线粒体通路诱导Hep G2发生凋亡的机制。方法分别采用MTT法、PI染色法、流式细胞术检测苦参碱对肝癌Hep G2细胞的增殖抑制、周期调控、诱导凋亡作用;采用Western Blot检测苦参碱诱导肝癌Hep G2细胞抑凋亡蛋白Bid、Bcl-2的表达量。结果苦参碱可抑制Hep G2细胞的增殖作用和诱导凋亡,呈时间和剂量依赖性,并可将Hep G2细胞周期阻滞在G0/G1期。苦参碱可引起线粒体膜电位崩溃,同时诱导Bid、Bcl-2表达下调,Bax表达上调。结论苦参碱通过调节细胞内Bid、Bcl-2和Bax蛋白的表达,经线粒体信号转导途径诱导人肝癌细胞系Hep G2发生凋亡。     

N-乙酰半胱氨酸萝卜硫素通过上调Bak和下调Bcl-2蛋白表达诱导前列腺癌细胞凋亡

目的探究N-乙酰半胱氨酸萝卜硫素(D,L-sulforaphane N-Acetyl-L-cysteine,SFN-NAC)诱导前列腺癌细胞凋亡的分子机制。方法将前列腺癌细胞DU145和PC-3分别培养在6孔板中,当细胞密度达到70%-80%时,分别随机分为对照组(不加SFN-NAC)和加药组(加入终浓度为30μmol/L的SFN-NAC),继续在37℃培养24 h。利用AnnexinⅤ异硫氰酸荧光素/碘化丙啶(FITC/PI)双染流式细胞术检测细胞凋亡程度;利用Western blot检测Bak、Bax、Bcl-2和cleaved Caspase-3蛋白表达水平。结果与对照组相比,加药组的细胞凋亡率增高(P<0.01)。加药组细胞Bak蛋白表达较对照组升高(P<0.05);Bax蛋白表达也有少量增加,但与对照组相比,无统计学差异(P>0.05);Bcl-2蛋白表达下降(P<0.05);Bax/Bcl-2比值增加(P<0.01);细胞cleaved Caspase-3表达水平升高(P<0.01)。结论 N-乙酰半胱氨酸萝卜硫素通过上调Bak、下调Bcl-2蛋白表达,活化Caspase-3,诱导前列腺癌细胞凋亡,说明N-乙酰半胱氨酸萝卜硫素通过线粒体途径发挥抗癌作用。     

姜黄油诱导人肝癌细胞凋亡及其机制

探讨姜黄油的抗肝癌作用及其作用机制,采用二甲基噻唑蓝比色法(MTT)检测肝癌细胞的活力,流式细胞仪检测细胞凋亡率以及线粒体膜电位的变化,Western blot方法检测凋亡相关蛋白Bax、Bcl-2、caspase-3、caspase-9以及细胞色素C的表达。结果发现,姜黄油能时间依赖性地抑制肝癌细胞的生长,降低线粒体膜电位并诱导其凋亡;升高细胞内Bax与Bcl-2的比值;诱导细胞色素C从线粒体中的释放并激活caspase-9和caspase-3。结果提示姜黄油可能通过线粒体途径诱导肝癌细胞的凋亡。     

α-茄碱通过ROS/线粒体途径促进鼻咽癌细胞凋亡的分子机制

目的：通过体外实验探讨α-茄碱对人鼻咽癌细胞CNE-2的作用及其潜在机制。方法：CCK-8法检测α-茄碱处理后CNE-2 细胞的活性;流式细胞术检测细胞凋亡;Real-time PCR和Western blot实验检测CNE-2细胞内Bax、Bcl-2等凋亡相关基因和蛋白的表达水平。Mito-SOX流式细胞术检测线粒体ROS水平的变化;透射电子显微镜观察细胞线粒体超微结构。结果：α-茄碱显著抑制细胞活性,诱导细胞凋亡,呈时间与剂量依赖性。与对照组相比,10 μmol/L α-茄碱作用于CNE-2细胞48 h后,细胞内Aif和Caspase-8的表达量显著增加（P ＜0.01）;Bcl-2表达和Bcl-2/Bax比值显著降低（P ＜0.05）。α-茄碱引起线粒体内ROS水平显著升高,透射电镜结果显示了CNE-2细胞中线粒体的损伤。结论：α-茄碱能抑制细胞增殖活性,引起ROS水平升高,进而导致线粒体损伤,最终通过线粒体途径诱导CNE-2细胞凋亡。     

双氢青蒿素诱导前列腺癌 PC-3 细胞凋亡及其机制研究

目的 研究双氢青蒿素对前列腺癌 PC-3 细胞的凋亡诱导作用,并探讨其可能的机制。方法 MTT 法测定不同浓度双氢青蒿素对 PC-3 细胞增殖的影响,流式细胞仪 (FCM)、透射电镜 (TEM) 观察双氢青蒿素对 PC-3 细胞的凋亡诱导作用,免疫组化 (SP) 检测双氢青蒿素对 PC-3 细胞内 Bax、Bcl-2 蛋白阳性表达的影响。结果 双氢青蒿素对 PC-3 细胞有明显增殖抑制效应,能诱导 PC-3 细胞凋亡且具有浓度-时间依赖性,导致 PC-3 细胞线粒体肿胀、核碎裂、凋亡小体形成;随着双氢青蒿素浓度的增加,Bcl-2 蛋白阳性表达降低,而 Bax 蛋白阳性表达增加。结论 双氢青蒿素可能通过上调 Bax,下调 Bcl-2 蛋白表达诱导前列腺癌 PC-3 细胞凋亡。     

鱼藤素对乳腺癌细胞MDA-MB-231线粒体通透性转换孔的作用

目的研究鱼藤素对乳腺癌细胞MDA-MB-231线粒体通透性转换孔的作用。方法MTT法检测鱼藤素对MDA-MB-231细胞的增殖抑制作用;AnnexinⅤ-FITC/PI双染流式细胞术检测细胞凋亡率;罗丹明123单染法观察线粒体膜电位变化;蛋白免疫印迹法检测细胞质中Cytc的表达;分光光度法检测caspase-3蛋白活性;Fluo-3/AM荧光指示剂检测细胞内钙离子浓度变化;RT-PCR和蛋白免疫印迹法检测Bcl-2和Bax mRNA及其蛋白表达。结果鱼藤素对MDA-MB-231细胞增殖具有明显的抑制作用,呈时效和量效依赖关系(P0.05);鱼藤素处理细胞后,细胞线粒体膜电位降低,细胞质内Cytc蛋白表达水平升高,caspase-3活性明显提高,与未处理组相比差异具有统计学意义(P0.01)。流式细胞术检测显示细胞凋亡率和细胞内钙离子浓度随鱼藤素浓度的增加而增多;RT-PCR和蛋白印迹检测结果发现鱼藤素能使Bcl-2 mRNA和蛋白表达减少,而Bax表达增加。结论鱼藤素对乳腺癌MDA-MB-231细胞的增殖抑制、诱导凋亡作用可能与细胞内钙超载,Bcl-2和Bax表达改变影响线粒体通透性转换孔开放,诱导线粒体膜电位降低及Cytc释放有关。     

小檗碱通过ROS及caspase通路对肝癌HepG2细胞凋亡的作用

目的 探究小檗碱(BBR)对肝癌HepG2细胞的增殖抑制及作用机制.方法 采用噻唑蓝(MTT)法检测不同浓度(0、10、20、40、80 μmol/L) BBR对肝癌HepG2细胞的抑制作用;采用Hoechst 33342染色观察处理后HepG2细胞形态学变化;用流式细胞仪检测HepG2细胞凋亡率;间接荧光标记法检测BBR对细胞内活性氧簇(ROS)水平的影响;JC-1染色法检测细胞内线粒体膜电位的变化;应用Western blot法检测凋亡相关蛋白Bax、caspase-3、Cleaved-caspase-3的表达变化.结果 MTT法显示BBR可呈剂量、时间依赖性抑制肝癌HepG2细胞活力(P＜0.05);Hoechst 33342染色发现细胞形态发生明显变化,0μmol/L BBR处理的细胞呈淡蓝色,处理组细胞核固缩,形成凋亡小体,细胞凋亡率与药物浓度呈正相关(P＜0.05).流式细胞术发现BBR能使细胞内ROS升高,线粒体膜电位水平下降.Western blot法显示随着药物浓度升高,Bax、Cleaved-caspase-3、cleaved-聚腺苷二磷酸核糖聚合酶表达增强,Bcl-2蛋白表达降低,凋亡相关蛋白caspase-3表达下降.结论 BBR在体外可能通过增加ROS、提高促凋亡蛋白Bax并激活caspase-3表达抑制肝癌HepG2细胞增殖并诱导细胞凋亡.     

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…