苯海拉明等有机胺类约物干扰MTT还原法的应用

目的:探讨苯海拉明等有机胺类药物对MTT还原法测定细胞活性准确性的影响.方法:采用原代新生大鼠皮层星形胶质细胞模型,以不同浓度的苯海拉明等有机胺类药物分别作用于星形胶质细胞24 h,以形态学观察、Hoechst 33342和PI核荧光双染法对比MTT染色结果.分析苯海拉明等对MTT还原法应用的影响.结果:苯海拉明(10-4 mol/L)、吡拉明(10-4 mol/L)和卓兰替丁(10-3 mol/L)作用于星形胶质细胞能显著增加甲瓒的生成,而核荧光双染法显示,这时星形胶质细胞的增殖和凋亡以及坏死情况并没有发生显著性改变.通过显微镜形态学观察发现,苯海拉明(10-4 moL/L)能显著减少星形胶质细胞膜上的针状晶体--甲瓒.结论:苯海拉明等有机胺类药物通过抑制甲瓒外排而促进胞内MTT被大量还原,从而影响MTT还原法测定细胞活性的准确性.     

MTT法实验条件的研究

本实验用2种体外传代培养的白血病细胞株(K_(562),HL_(60))对MTT实验条件进行研究。结果表明:MTT代谢产物甲 分细胞数量呈线性关系。最佳实验条件为:实验用细胞数为2×10~5/孔或4×10~5/孔,MTT加入量为100μg(5mg/ml,20μl);其它实验条件为选用570nm为主波长,630nm为参考波长;96孔板不离心;MTT作用4小时。MTT法经济、准确、快速和简便,从而为临床急性白血病的药物敏感检测奠定方法学基础。     

耐药人肝癌细胞模型-7721/Adm的建立

目的　培养建立耐药肝癌细胞模型 - 772 1/ Adm并研究其生物学特性 .方法　应用人肝癌细胞株 - 772 1(以下简称772 1细胞 ) ,采用阿霉素 (Adm )大剂量间歇诱导法 ,建立耐药人肝癌细胞模型 - 772 1/ Adm (以下简称 772 1/ Adm ) .观察该细胞的生长规律 ;用 MTT法鉴定该耐药细胞模型对多种抗癌药物的多药耐药性 ;以流式细胞技术检测该耐药细胞模型细胞周期分布、细胞表面多药耐药基因 (MDR)的表达产物 - P-蛋白 (P- gp)、多药耐药相关蛋白 MRP及谷胱甘肽硫转移系统(GSH/ GST)的表达等 .结果　 1经 MTT法鉴定 772 1/ Adm细胞较 HCC- 772 1细胞的阿霉素半数致死浓度 (IC5 0 )增大4.6 5倍 ,并对多种抗癌药物产生耐药性 ,其耐药性提高了 2～6倍不等 ;2耐药细胞倍增时间明显延长 ,S期细胞减少 ,而G2期细胞增多 ;3该耐药模型 P- gp,MRP表达有非常显著的升高 (P<0 .0 1) :P- gp表达为 94.6 % ,MRP表达为 6 9.4% ,而 GSH/ GST的表达无明显变化 (P<0 .0 5 ) .结论　 1HCC-772 1/ Adm是一个明确的多药耐药细胞模型 ;2 772 1/ Adm细胞具有耐药细胞的基本生物学特性     

耐长春新碱肝癌细胞系HCC-7721/VCR的建立

目的: 培养建立耐长春新碱肝癌细胞系HCC-7721/VCR,并对其生物学特性进行研究.方法: 应用人肝癌细胞系HCC-7721,采用长春新碱大剂量间歇诱导法,建立耐药人肝癌细胞系HCC-7721/VCR.观察该细胞的生长规律;用MTT法鉴定该耐药细胞模型对多种抗癌药物的多药耐药性;以流式细胞技术检测该耐药细胞模型细胞周期分布.结果: 经MTT法鉴定,HCC-7721/VCR细胞较HCC-7721细胞的长春新碱半数致死浓度(IC50)增大了4.15倍,并对多种化疗药物产生耐药性,其耐药性提高了2～4倍不等;耐药细胞倍增时间明显延长,S期细胞减少,而G1、G2期细胞增多.结论: HCC-7721/VCR是一个明确的多药耐药细胞模型,具有耐药细胞的基本生物学特性.     

瘦素对结肠癌细胞株HT-29的影响

目的:观察瘦素对结肠癌细胞株HT-29的细胞生长的作用。方法:贴壁培养HT-29细胞,分别给予不同浓度的瘦素(0.5×10-6、5×10-5,1×10-4,2×10-4g/L)。并同时以浓度为1×10-4g/L的表皮生长因子(EGF)作为阳性对照组,通过测定生长曲线、四甲基谷氮唑蓝实验MTT法观察瘦素对HT-29细胞生长的影响。结果:通过生长曲线,发现在相同时间段,瘦素浓度在5×10-6-2×10-4g/L时,呈剂量依赖性促进HT-29细胞增殖。但即使在瘦素促细胞增殖最为明显的2×10-4g/L组中,和阳性对照组EGF比较,其细胞增殖数量仍为低。MTT法结果显示,不同浓度的瘦素作用于HT-29细胞后,在72 h内,呈时间、剂量依赖性促进HT-29细胞生长,其增长率在5.1%-67.9%之间。细胞计数和MTT法均显示瘦素可促进结肠癌细胞HT-29的生长和增殖。提示瘦素作为多功能的细胞因子和结肠癌的发生密切相关。结论:在离体实验中,瘦素具有促结肠癌细胞HT-29生长增殖的作用,且和其浓度、时间相关。     

卵巢癌个体性化疗方案的初步研究

目的评价卵巢癌细胞对8种化疗药物的敏感性,为临床卵巢癌患者个体性化疗方案的制定提供有用的研究模式.方法①应用MTT法检测8种化疗药物对卵巢癌细胞系HO-8910细胞和对照组不同时间、不同浓度化疗药物及其组合的细胞抑制率,筛选出抑制率最高的化疗药物并进行组合.②通过FCM法检测其诱导细胞凋亡的效果,采用Annexin-V-FITC结合PI染色和TdT介导X-dUTP缺口末端标记(TUNEL)法.结果①在MTT法定量检测中,8种化疗药物作用48h对HO-8910细胞系抑制率与对照组相比具有不同程度的差异性,其中以4PPC(32μg/ml)的CDDP与4PPC(0.12μg/ml)的As2O3效果最显著,此时H0-8910细胞系的抑制率分别为80.43%和73.37%,与对照组比较,差异显著(P=0.01)(2).MTT法定量检测21种化疗药物组合对HO-8910细胞系抑制率,筛选出抑制率最高的化疗药物组合:CDDP BLM 紫杉醇(抑制率94.44%),与对照组比较,差异显著(P=0.0125).(3).FCM法定量检测组合CDDP BLM 紫杉醇作用2、4、6、8、10、12、24、48h 8个时间段对HO-8910细胞系的细胞凋亡率,与空白对照组比较有不同程度的差异,其中以作用48h的细胞凋亡率最高(75.70%),差异显著(P=0.0136).结论①卵巢癌细胞的化疗敏感性与化疗药诱导的卵巢癌细胞凋亡密切相关.②细胞抑制率,早期细胞凋亡,晚期细胞凋亡,细胞凋亡率可作为制订卵巢癌个体性治疗方案的评价指数体系.③在细胞抑制率的基础上优选出相应的抗癌药物,再通过细胞凋亡率评价抗癌药物的优化组合方案,这一肿瘤个体性化疗方案制定程序具有可行性,为卵巢癌者个体性化疗方案的制定提供了一个有用的研究模式.     

H_2O_2诱导RINm5F细胞凋亡及升清降糖合剂的保护作用

[目的]升清降糖合剂(SQ)是临床治疗糖尿病的复方,为研究其治疗机制,通过其作用于H2O2致胰岛细胞凋亡模型,了解复方对胰岛细胞的保护作用,为临床治疗糖尿病提供实验依据。[方法]以H2O2诱导建立胰岛细胞凋亡模型;采用半仿生提取法提取复方有效成分;以不同浓度(5、10、50μg.mL-1)作用于胰岛细胞凋亡模型,光镜下观察细胞形态学改变,MTT法比较各组凋亡与增殖率,并检测各组上清及胞内SOD、MDA浓度。[结果]以半仿生提取法成功提取复方有效成分;以不同时间和不同浓度干预细胞后,经MTT法检测和AO/PI形态学观察,确定凋亡模型条件为0.2mmol.L-1 H2O2干预24h;以不同浓度药物干预凋亡模型24h后,细胞形态明显好转,经MTT法检测细胞活性,10μg.mL-1药物组明显提高细胞活性(P<0.01);SQ能提高胞内SOD活性和SOD/MDA比例(P<0.05),同时降低上清中MDA水平(P<0.05)。[结论]升清降糖合剂对H2O2致胰岛细胞凋亡有保护作用,其作用机制与提高SOD/MDA比例有关。     

索拉非尼逆转肝癌细胞多药耐药的实验研究

目的 探讨索拉非尼体外对人肝癌细胞(BEL-7402/FU)多药耐药性的逆转作用及可能机制.方法 以MTT比色法测定索拉非尼对肝癌细胞的剂量效应曲线,并以流式细胞仪检测索拉非尼对肝癌细胞内罗丹明123 (Rho123)浓度的影响,根据上述实验结果选取合适的索拉非尼实验剂量.以MTT法测定索拉非尼对抗癌药物细胞毒性的影响,用流式细胞仪检测细胞膜转运蛋白(P-gp)的表达,RT-PCR法检测mdr1基因的表达.结果 索拉非尼浓度在4μmol/L时逆转效率较高且毒副作用较小,4 μmol/L的索拉非尼能部分逆转BEL-7402/FU细胞的耐药性,对ADM、5-FU、GEM、DDP的逆转倍数分别为2.98、7.16、1.99、10.08倍,并可部分下调BEL-7402/FU细胞P-gp的表达,使mdrl基因表达与对照组相比下降了27.3%.结论 索拉非尼具有体外逆转人肝癌细胞多药耐药性的作用,可能与下调mdr1基因表达、增加细胞内化疗药物的蓄积有关.     

用紫外与可见光分光分析仪测定PASMC增殖的MTT比色法

为使 MTT比色法得到更广泛的应用 ,对紫外与可见光分光分析仪测定细胞增殖 MTT比色法的实验条件进行了探索。结果发现 :以 5 mg/ m l MTT在 0 .1× 10 4～ 2 .4× 10 4细胞范围内进行检测 ,可获得较好的线性检测结果 (γ=0 .9872 )。与酶标仪直接测定比较 ,具有灵敏度高、数据准确、检测范围广 ,值得推广应用。     

MTT显色法抗癌药物敏感性试验的适用性研究

本文对在96孔细胞培养板中短期培养下的人红白血病K_(562)细胞株、小鼠肠癌CT_(26)实体瘤细胞、小鼠S_(180)实体瘤细胞、人大肠癌手术切除标本肿瘤细胞及人急性白血病细胞的存活适应状况,用计数及MTT显色法进行了研究.结果表明,MTT法对K_(562)细胞及人急性白血病细胞的体外抗癌药物抑瘤试验结果稳定,柔红霉素、高三尖杉酯碱等药物对K_(562)细胞及部分人急性白血病细胞显示较高的敏感性.因而,该法对人急性白血病有效治疗药物的临床选择有一定价值.但MTT法尚难适用于实体肿瘤的抗癌药物敏感性试验.     

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…