miR-520a-5p通过调控有氧糖酵解诱导前列腺癌细胞多西他赛耐药

目的：探索mi R-520a-5p对前列腺癌增殖能力和多西他赛耐药性的影响及其作用机制。方法：使用RT-q PCR检测PC-3/DU145前列腺癌细胞及RWPE-1前列腺正常上皮细胞中mi R-520a-5p表达水平;使用免疫印迹检测mi R-520a-5p是否调控糖酵解关键酶己糖激酶2(hexokinase 2,HK2)和磷酸果糖激酶(phosphofructokinase,PFKM)表达水平;使用CCK-8实验检测mi R-520a-5p对前列腺癌细胞增殖能力和多西他赛耐药性的影响;使用ELISA评估培养上清中葡萄糖消耗水平和乳酸生成水平。依据转染物的不同,将所有细胞分成4组：minic组、minic NC组、inhibitor组、inhibitor NC组。结果：mi R-520a-5p在前列腺癌细胞中表达下调(P<0.001)。与minic NC组比较,minic组可显著降低PC-3和DU145前列腺癌细胞的增殖能力(P<0.001)、多西他赛半抑制浓度(IC₅₀)、葡萄糖消耗水平和乳酸生成水平(P<0.001);与inhibitor N...     

20(S)-原人参二醇对人前列腺癌细胞凋亡的诱导作用

目的:研究20(S)-原人参二醇对人前列腺癌细胞凋亡的影响。方法:采用台盼蓝染色、噻唑蓝(MTT)法、Hochest33258染色和DNA Ladder方法分别测定20(S)-原人参二醇对DU145和PC3细胞凋亡诱导作用。结果:20(S)-原人参二醇可明显降低DU145和PC3细胞存活率和细胞活性,IC50分别为38.0μmol/L和28.6μmol/L。20(S)-原人参二醇处理后DU145和PC3细胞核出现典型的凋亡形态学改变,琼脂糖电泳呈现明显的DNA Ladder条带。结论:20(S)-原人参二醇可有效诱导人前列腺癌DU145和PC3细胞的凋亡并有剂量依赖关系。     

多西紫杉醇联合二烯丙三硫协同诱导前列腺癌细胞凋亡的研究

【目的】探讨多西紫杉醇联合二烯丙三硫(DATS)对前列腺癌PC-3细胞的协同杀伤效应及其机制。【方法】PC-3细胞接种96孔板,分别给予多西紫杉醇(5～60 nmol/L)和DATS(5～60μmol/L)单药处理24、48、72 h;联合用药采用15μmol/L DATS联合不同浓度多西紫杉醇(5～60 nmol/L)处理48 h。MTT法检测多西紫杉醇和(或)DATS对PC-3细胞生长的影响。DAPI染色、流式细胞仪检测多西紫杉醇、DATS及二者联合对PC-3细胞凋亡和细胞周期的影响;Western-blot测定Bcl-2、Bax、Bcl-xL/xS及细胞色素C、caspase-9、caspase-3的表达及活性变化。【结果】多西紫杉醇、DATS均能抑制PC-3细胞的生长,DATS可明显提高多西紫杉醇的杀伤效应,DATS(15μmol/L)联合多西紫杉醇(10 nmol/L)作用48 h的细胞生存率为(50.1±1.0)%,低于多西紫杉醇(67.4±1.0)%和DATS(69.2±1.8)%,二者具有协同抑瘤作用;联合组使细胞凋亡率及G2/M期细胞比率明显增高,并更能显著抑制Bcl-2、Bcl-xL的表达,而且联合组使细胞色素C的释放及caspase-9、caspase-3的裂解活化时间提前了8 h。【结论】多西紫杉醇及DATS共同诱导细胞凋亡是二者发挥协同效应抑制前列腺癌细胞生长的重要机制,其可能与细胞周期阻滞、Bcl-2、Bcl-xL的表达降低、细胞色素C的释放及caspase-9、caspase-3的活性变化有关。     

辛伐他汀对前列腺癌PC-3细胞影响的实验研究

目的:观察辛伐他汀对前列腺癌PC-3细胞增殖、凋亡的影响,探讨辛伐他汀对前列腺癌的作用。方法:MTT比色法检测PC-3细胞的增殖;Hoechst 33258染色后荧光显微镜法观察PC-3细胞凋亡的形态学改变,并计算凋亡核百分率;Annexin V-FITC/PI双染色,流式细胞术检测细胞凋亡。结果:(1)辛伐他汀可显著抑制PC-3细胞的体外增殖,并呈浓度和时间依赖性(P<0.05)。(2)Hoechst 33258染色显示PC-3细胞经辛伐他汀(20?mol/L)分别处理24h和48h后,细胞出现凋亡,形态学变化表现为凋亡细胞体积缩小,核固缩,镜下细胞核呈致密浓染或碎块状致密浓染。FACS检测结果显示,20?mol/L辛伐他汀处理12h、24h、48h后的凋亡细胞数分别为14.81±1.08%、23.42±2.33%、40.77±4.06%。PC-3细胞分别经10μmol/L、20μmol/L、40μmol/L辛伐他汀处理48h后,凋亡细胞数逐渐增多,凋亡核百分率计数分别为(11.20±3.33)%,(31.20±3.08)%,(56.24±4.50)%。FACS检测结果显示,10μmol/L、20μmol/L、40?mol/L辛伐他汀处理PC-3细胞48h后的凋亡细胞数分别为17.26±1.44%、32.45±2.56%、60.47±3.98%。结论:(1)辛伐他汀能明显抑制人前列腺癌PC-3细胞的体外增殖。(2)辛伐他汀可诱导人前列腺癌PC-3细胞发生凋亡。     

甘草甜素诱导前列腺癌细胞株DU145凋亡的体外研究

目的:探讨甘草甜素(GL)诱导前列腺癌细胞株DU145凋亡的作用机制。方法:采用不同浓度的GL处理细胞后,用流式细胞仪检测前列腺癌细胞株DU145的凋亡率及细胞周期分布。结果:50、100μmol/L浓度的GL能诱导前列腺癌细胞DU145凋亡,凋亡率与剂量成正相关;DU145随药物浓度及作用时间变化,细胞周期分布呈现G1期细胞比例逐渐增高,并出现典型的凋亡峰。结论:甘草甜素能诱导前列腺癌细胞凋亡,并明显抑制癌细胞增殖,具有直接抗肿瘤作用。     

转移性去势抵抗性前列腺癌化疗与激素治疗策略

以多西他赛为核心的化疗方案已经成为转移性去势抵抗性前列腺癌（ metastatic castrate-resistant prostate cancer,mCRPC）治疗的一线方案。但在过去的几年中,一些新药的出现改善了患者的总体生存,同时也使优化个体化治疗方案成为可能。已有证据表明阿比特龙、卡巴他赛、镭-223、sipuleucel-T、恩杂鲁胺等与多西他赛一同使去势抵抗性前列腺癌患者生存获益。另外,mCRPC患者对多西他赛一线治疗初始反应良好,出现疾病进展后复治应作为一线治疗的延伸。这些新的治疗手段使mCRPC治疗方案更加复杂化,并且使以往的序贯治疗方案向基于一定规则的新的治疗方式转变。     

雄激素对耐多西他赛前列腺癌细胞P-gp、GST-π表达的影响

目的 研究雄激素对耐多西他赛前列腺癌细胞P-糖蛋白（P-gp）、谷胱甘肽-S-转移酶-π（GST-π）表达的影响.方法 人前列腺癌细胞株LNCaP及PC-3在含多西他赛的培养液中诱导耐药细胞株（LNCaP/Docetaxel,PC-3/Docetaxel）,将耐药细胞株分别在含有二氢睾酮（DHT）培养液和含二氢睾酮＋比卡鲁胺培养液中培养7 d.免疫细胞化学法检测两组细胞P-gp、GST-π的表达水平.结果 经DHT诱导后,P-gp和GST-π在LNCaP/Docetaxel及PC-3/Docetaxel细胞系的表达明显升高（χ2＝7.367,6.874,P＜0.05）;而经DHT＋比卡鲁胺联合作用后,P-gp和GST-π的表达保持不变（P＞0.05）.结论 二氢睾酮可诱导耐多西他赛前列腺癌的耐药相关蛋白P-gp及GST-π增量表达,雄激素受体阻断剂比卡鲁胺可阻断二氢睾酮的诱导作用.     

AG-490诱导人前列腺癌细胞凋亡的研究

目的 研究抑制Janus kinase-signal transducer and activator(JAK-STAT)途径的磷酸化，诱导前列腺癌细胞凋亡的效应。方法 不同浓度的JAK2磷酸化抑制剂AG-490处理人前列腺癌PC-3M细胞，MTT比色法和流式细胞术检测半数细胞生长抑制剂量(IC50)和细胞凋亡率，透射电镜和TUNEL法观察凋亡形态。结果 AG-490能够抑制PC-3M细胞增殖并表现为剂量依赖性，48h的IC50为56．8μmol／L；用AG-490处理PC-3M细胞后可观察到典型的细胞凋亡形态。结论 JAK-STAT途径磷酸化在支持PC-3M细胞增殖过程中起重要作用，AG-490能够有效抑制前列腺癌PC-3M细胞增殖并促进其凋亡。     

FGF8b调控前列腺癌细胞上皮间质转化的分子机制

目的:探讨成纤维细胞生长因子8b(fibroblast growth factor 8b,FGF8b)促进前列腺癌DU145细胞上皮间质转化的分子机制。方法:先选取3组细胞,分别为空白对照组(DU145细胞)、阴性对照组[(DU145细胞转染空白质粒(简称pcDNA3.1/DU145)]和实验组[DU145细胞转染FGF8b(简称FGF8b/DU145)]。采用Western印迹检测细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)通路活性,然后将FGF8b/DU145细胞及DU145细胞在PD98059(ERK激酶抑制剂)作用下分为4组:A组为2%胎牛血清(FBS)处理的FGF8b/DU145细胞;B组为2%FBS+PD98059(50μmol/L)处理的FGF8b/DU145细胞;C组为2%FBS处理的DU145细胞;D组为2%FBS+PD98059(50μmol/L)处理的DU145细胞,培养观察细胞形态学的变化,最后Western印迹和Transwell小室检测各组细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)相关标志物的变化以及细胞的迁移能力改变。结果:实验组ERK1/2活性明显高于空白对照组和阴性对照组。给予PD98059后,B组与未给予PD98059组的A组相比,上皮细胞标志物上皮钙黏素蛋白表达量明显上调(P<0.05);间质标志物波形蛋白表达量明显下调(P<0.05)。细胞迁移试验结果提示:在给予PD98059后,B组FGF8b/DU145细胞的迁移能力与A组相比明显减弱。结论:FGF8b调控前列腺癌细胞DU145发生上皮-间质转化,可能由ERK激酶通路介导,MAPK激酶1作为该信号通路的关键因子之一,可能是干预前列腺侵袭转移的有效靶点。     

冬凌草甲素通过抑制Akt通路诱导人雄激素非依赖性前列腺癌细胞DU145细胞凋亡

目的:观察冬凌草甲素对人雄激素非依赖性前列腺癌细胞DU145细胞增殖、凋亡的影响,并分析可能的作用机制。方法:体外培养的DU145细胞,用不同浓度冬凌草甲素(浓度0、10、20、40、80、100μmol/L)处理24、48、72 h,MTT法测定其对DU145细胞增殖的影响;冬凌草甲素(浓度分别为0、20、40、80μmol/L)处理DU145细胞48 h后,采用流式细胞仪检测细胞凋亡,Western blot法检测Akt、p-Akt、p-GSK-3β、Bax及Bcl-2蛋白的表达情况。结果:冬凌草甲素对DU145细胞的增殖具有抑制作用,且呈剂量、时间依赖性;DU145细胞经冬凌草甲素作用48 h后,细胞凋亡率显著增加;Bax表达上调,而Bcl-2表达下调;p-Akt及其下游蛋白p-GSK-3β的表达降低。结论:冬凌草甲素可通过抑制DU145细胞增殖并促进其凋亡,其作用机制可能与抑制Akt信号通路激活,继而上调Bax并下调Bcl-2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…