AAV介导的BATF2/SARI过表达抑制肝癌细胞HepG2增殖

目的探讨腺相关病毒(adeno-associated virus,AAV)介导的BATF2/SARI过表达对肝癌细胞HepG2细胞活力的影响及其分子机制。方法通过分子克隆技术构建AAV介导的BATF2/SARI过表达载体,用包装纯化后的AAV-BATF2感染HepG2细胞,MTT检测其对细胞活力的作用,荧光素酶分析其对NFκB转录活性的影响。结果重组AAV-BATF2载体构建成功,AAV-BATF2感染的HepG2细胞生长活力受到明显抑制,NFκB转录活性明显下调。结论重组AAV-BATF2病毒感染人肝癌细胞HepG2可抑制肿瘤细胞增殖,其对NFκB转录活性的抑制可能是其发挥作用的机制之一。     

大麻受体激动剂THC抑制肝癌细胞HepG2增殖和诱导其凋亡的实验研究

目的 探讨激活大麻受体对肝癌细胞增殖和凋亡的作用,并对其机制进行初步研究.方法 将HepG2细胞分成对照组、不同剂量大麻受体激动剂delta9-tetrahydrocannabinol(THC)处理组.MTT法测定THC对HepG2 细胞增殖的影响;DNA梯度电泳法和流式细胞仪检测分析各组细胞凋亡情况;Western blot法分析细胞大麻受体CB1、CB2、Caspase 3和c-myc的蛋白表达;分光光度法检测Caspase 3的活性.结果 HepG2细胞大麻受体CB1、CB2表达较L02正常肝细胞增高.THC能抑制HepG2细胞增殖,诱导HepG2细胞凋亡,上述效应具有剂量依赖性.THC可抑制HepG2细胞c-myc的表达,诱导HepG2细胞Caspase 3的蛋白表达和活性.结论 大麻受体激动剂THC能抑制肝癌细胞增殖,诱导肝癌细胞凋亡,此效应可能与其影响c-myc的表达和Caspase 3的活性相关.     

Quercetin对HepG2细胞的抑制作用及PDGFR-β的表达

目的 探讨植物化学物质槲皮素(Quercetin)对人类肝癌细胞HepG2的抑制作用和血小板衍生生长因子β受体(PDGFR-β)表达的关系。方法 采用免疫组化SABC法观察Quercetin抑制HepG2中PDGFR-β表达,并采用透射电镜观察Quercetin作用后。HepG2细胞的凋亡情况。结果 细胞增殖率检测表明Quercetin可以抑制HepG2细胞的增殖,且呈剂量依赖性;Quercetin可以抑制HepG2细胞PDGFR-β的表达,且呈剂量依赖性;透射电镜下可见凋亡小体。结论 槲皮素对HepG2细胞增殖有明显抑制作用,其机制可能是诱导PDGFR-β的增加,再通过PDGFR-β介导的细胞内信号体系控制细胞凋亡而实现的。     

白藜芦醇通过下调microRNA-151表达抑制肝癌细胞株HepG2细胞活性的研究

目的 研究白藜芦醇(Res)对肝癌细胞株HepG2宿主基因黏着斑激酶(FAK)和内含子miR-151表达的影响,探讨其抗肿瘤的可能作用机制.方法 分别以50、100、150、200 μmol/L Res作用于体外培养的HepG2细胞24、48和72 h.CCK-8法检测细胞增殖;Real-Time PCR检测细胞FAK mRNA和miR-151表达.利用miR-151模拟物转染获得miR-15过表达HepG细胞株(miR-151过表达组),流式细胞仪和Hoechst 33342染色观察过表达miR-151对HepG2细胞周期及凋亡的影响.以转染模拟物突变体细胞作为阴性对照,未转染细胞作为空白对照.结果 Res对体外培养HepG2细胞的增殖活性及FAK mRNA和miR-151表达具有显著抑制作用,并在一定范围内呈现浓度和时间依赖性.miR-151过表达组HepG细胞miR-151表达明显高于阴性对照组和空白对照组(P＜0.001);流式细胞仪检测和Hoechst 33342染色观察结果显示,miR-15过表达使HepG细胞G0/G1期缩短,细胞周期进程加快,并能抑制细胞凋亡.结论 Res可能通过下调miR-151表达,抑制肝癌细胞HepG2增殖并诱导凋亡.     

HMGB1对人肝癌细胞株HepG2体外增殖的影响

目的：观察HMGB1对人肝癌细胞株HepG2增殖的影响并初步探讨其可能机制。方法：采用不同剂量(0,10,50,100 ng/mL)重组HMGB1(rHMGB1)刺激HepG2细胞24 h,用MTT法检测HepG2细胞增殖,Western印迹和RT PCR法检测cyclin D1,PCNA的表达。结果：不同剂量(10,50,100 ng/mL)rHMGB1组HepG2细胞增殖能力均明显高于对照组（P＜0.05）。HMGB1抗体（20 μg/mL）能中和HMGB1(10ng/mL)对HepG2细胞的促增殖作用（P＜0.05）。Western印迹和RT PCR显示,与对照组比较,HMGB1能明显促进HepG2细胞PCNA,cyclin Dl蛋白和mRNA的表达（P＜0.05）,HMGB1抗体能明显抑制HMGB1上调PCNA,cyclin Dl蛋白和mRNA的表达（P＜0.05）。结论：HMGB1可能通过上调cyclin Dl和PCNA表达,促进HepG2细胞增殖。HMGB1抗体对肝细胞癌可能有治疗作用。     

槲皮素对人肝癌细胞生长、迁移能力的影响及机制研究

目的探讨槲皮素对人肝癌细胞(HepG2)生长增殖及迁移能力的影响及机制。方法培养对数生长的HepG2细胞,MTT试验分析槲皮素对HepG2生长的影响,划痕实验观察加入槲皮素前、后HepG2迁徙能力的变化。Western blot检测其中细胞迁移黏附因子(IQGAP1)表达的变化。结果加入槲皮素后,HepG2细胞逐渐变圆、脱落,增殖变慢;MTT实验显示HepG2细胞的生长抑制效应与槲皮素有浓度、时间依存关系;Western blot显示加入槲皮素后HepG2细胞中IQGAP1表达明显降低。结论槲皮素能明显抑制HepG2细胞生长增殖,减弱其迁移能力,从而达到抑制恶性细胞增殖转移的作用,这种能力可能是通过抑制IQGAP1表达实现的。     

姜黄素抑制肝癌细胞HepG2增殖及干细胞标志物表达

目的:探讨姜黄素能否抑制肝癌细胞HepG2的增殖及干细胞标志物的表达。方法:不同剂量梯度的姜黄素(0、20、40和60μmol/L)作用肝癌细胞系HepG2 48 h后,MTT实验和克隆形成实验检测HepG2的增殖情况,划痕实验检测其迁移情况,成球实验检测姜黄素对HepG2干细胞的抑制情况,Real-time PCR检测姜黄素处理HepG2干细胞标志物CD133的表达情况,Western blot检测干细胞转录因子OCT4的表达情况。结果:姜黄素可抑制肝癌细胞HepG2的增殖[60μmol/L剂量时抑制率达(68.12±3.09)%],并抑制其克隆形成和迁移能力;姜黄素还可抑制HepG2的成球能力;同时可导致HepG2干细胞标志物CD133的表达降低[60μmol/L剂量时下降率达(48.36±2.04)%],并导致干细胞相关转录因子OCT4的表达降低。结论:姜黄素通过抑制肿瘤干细胞从而降低肝癌复发和转移,有望成为肝癌治疗的辅助性药物。     

抗肿瘤血管生成药bevacizumab对VEGF促人肝癌细胞株HepG2增殖的阻断作用

目的:观察bevacizumab对肝癌细胞株HepG2增殖的影响,并探讨其可能的作用机制.方法:应用免疫细胞化学法和RT-PCR分别从蛋白和基因水平检测肝癌细胞株HepG2中血管内皮生长因子(VEGF)及其受体Flt-1、KDR的表达;ELISA检测HepG2培养上清液中VEGF蛋白的浓度;人重组VEGF(rhVEGF)和bevacizumab分别处理HepG2细胞后,MTT法检测细胞增殖活性,应用RT-PCR和Western印迹分别从基因和蛋白水平检测VEGF表达量的变化.结果:HepG2细胞中VEGF、Flt-1和KDR蛋白均呈阳性表达.rhVEGF可促进HepG2细胞的增殖,在0~100 ng/ml区间其促进增殖的作用呈剂量依赖性;而bevacizumab可抑制HepG2细胞的增殖,浓度为0.1、1、10、20 μg/ml时细胞增殖抑制率分别为(8.76%±1.15)%、(26.83±1.20)%、(31.87±1.30)%、(28.20±1.28)%.rhVEGF可促进HepG2细胞中VEGF的表达,而bevacizumab能抑制VEGF的表达.结论:Bevacizumab可能通过阻断VEGF的促增殖作用而抑制HepG2细胞增殖.     

白藜芦醇对肝癌Bel-7404细胞增殖、凋亡及侵袭的影响

目的 研究白藜芦醇对肝癌Bel-7404细胞增殖、凋亡及侵袭的影响.方法 MTT法和增殖细胞核抗原(PCNA)测定法检测白藜芦醇对Bel-7404细胞增殖的影响;流式细胞术和TdT酶介导的dUTP缺口末端标记法检测白藜芦醇对Bel-7404细胞凋亡的影响;黏附实验、运动实验和侵袭实验检测白藜芦醇对Bel-7404细胞侵袭能力的影响.结果 白藜芦醇浓度＞25μmol/L可抑制肝癌细胞Bel-7404增殖,并诱导凋亡(P＜0.01);浓度＞100 μmol/L正常肝细胞L02的增殖也受到抑制,并产生凋亡(P＜0.01).25,50μmol/L的白藜芦醇可抑制Bel-7404细胞黏附细胞外基质纤连蛋白(FN)及降解基质的能力(P＜0.01),对该细胞的运动能力无明显抑制作用.结论 一定浓度的白藜芦醇可抑制肝癌Bel-7404细胞增殖,诱导其凋亡,并可能通过抑制Bel-7404细胞与细胞外基质FN的黏附及对基质的降解来抑制Bel-7404细胞的侵袭能力;在较高浓度下(≥100 μmol/L),白藜芦醇对正常肝细胞株也产生抑制增殖和诱导凋亡的毒性作用.     

白藜芦醇体外对肝癌HepG2细胞分化及P21WAF1/CIP1表达的影响

目的 探讨白藜芦醇在体外对肝癌细胞分化及相关周期蛋白依赖激酶抑制因子P21WAF1/CIP1的影响.方法 体外培养肝癌HepG2细胞,用MTT法检测白藜芦醇对HepG2细胞的生长抑制作用,用倒置显微镜观察肝癌细胞的形态改变.用放射免疫法检测其AFP分泌,以RT-PCR方法检测HepG2细胞中P21WAF1/CIP1 mRNA的表达,用免疫细胞化学检测其P21WAF1/CIP1蛋白的表达.结果 白藜芦醇呈时间剂量性抑制HepG2细胞株的增殖,使其亚细胞结构趋于正常,AFP分泌量下降,并显著上调HepG2细胞中P21WAF1/CIP1 mRNA和蛋白的表达.结论 白藜芦醇能诱导HepG2细胞在体外向正常肝细胞分化,并上调其P21WAF1/CIP1的表达.     

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…