纳米二氧化钛颗粒体外对人肺腺癌A549细胞的毒性效应

目的探讨不同作用时间下粒径为25nm的锐钛矿型二氧化钛(TiO2)颗粒对人肺腺癌A549细胞的毒性效应。方法将100μg/ml的纳米TiO2颗粒悬液与A549细胞在37℃、5%CO2的细胞培养箱中分别孵育1h、4h、8h和24h后,收集细胞。通过乳酸脱氢酶(LDH)检测实验检测纳米TiO2颗粒对细胞膜的损伤情况;通过ATP检测实验检测纳米TiO2颗粒对线粒体产生的影响;通过超氧化物歧化酶(SOD)活性检测实验检测纳米TiO2颗粒对细胞的氧化损伤情况;采用透射电子显微镜观察纳米TiO2颗粒引起细胞超微结构的变化;利用CCK-8法检测纳米TiO2颗粒对细胞存活率的影响。结果随着纳米TiO2颗粒作用时间的延长,细胞外液中LDH活性增强,细胞内ATP浓度降低,SOD活性降低,细胞存活率呈现明显下降(P<0.05),且细胞线粒体和内质网出现不同程度的肿胀和扩张。结论纳米TiO2颗粒体外能够引起肺腺癌细胞氧化损伤,抑制细胞生长,且对细胞的毒性效应存在时间依赖性。     

纳米二氧化硅颗粒对血管内皮细胞的毒性及其凋亡诱导作用

目的:研究纳米二氧化硅(SiO₂)颗粒的血管内皮细胞毒性和可能的作用机制,为探讨纳米SiO₂颗粒毒性效应及安全性评价提供参考依据。方法:将体外培养的人脐静脉内皮细胞(HUVECs)随机分为对照组和不同浓度纳米SiO₂颗粒暴露组,暴露浓度分别为12.5、25.0、50.0及100.0 mg·L^-1。采用透射电镜(TEM)观察纳米SiO₂颗粒粒径、形貌及分散性。细胞处理24 h后,电感耦合等离子体原子发射光谱(ICP-AES)法测定细胞中硅水平;MTT法测定细胞活力;乳酸脱氢酶(LDH)释放法检测细胞膜的完整性;DCFH-DA荧光探针标记激光共聚焦显微镜观察细胞中活性氧(ROS)水平;DAPI染色荧光显微镜下观察细胞核形态;Annexin Ⅴ/PI双染标记流式细胞术(FCM)检测细胞凋亡率;罗丹明123线粒体试剂盒检测细胞线粒体膜电位。结果:TEM观察,纳米SiO₂颗粒分布均匀,大小一致,颗粒呈球形,分散性好,未发生聚集。用Image J 软件计算得到颗粒平均粒径为(57.66 ± 7.30)nm。与对照组比较,纳米SiO₂颗粒作用于HUVECs后,细胞活力下降(P<0.05),细胞中硅水平和培养液中LDH活性增加(P<0.05),细胞中ROS水平升高,线粒体膜电位下降,甚至发生细胞凋亡。上述细胞效应均表现为随颗粒作用浓度的增加而逐渐明显,呈现一定的剂量依赖关系。结论:纳米SiO₂颗粒具有血管内皮细胞毒性,可诱导ROS生成和氧化应激,从而破坏细胞膜、损伤线粒体,最终引发细胞凋亡。     

5 nm金纳米颗粒对大鼠皮质神经元的作用及机制初探

目的:探讨直径5 nm的小尺寸金纳米颗粒(gold nanoparticles,GNPs)对大鼠皮质神经元细胞的作用及相关机制?方法:使用硼氢化钠还原法制备GNPs,用透射电镜(TEM)及紫外-可见光光谱(UV-Vis)进行鉴定?用5% 牛血清白蛋白(BSA)对GNPs进行包裹提纯后制备成不同浓度(600?1 200?2 400 μg/L)的GNPs培养液?采用免疫荧光染色鉴定原代培养的大鼠皮质神经元纯度?体外培养原代大鼠皮质神经元72 h后,以正常培养的神经元为对照组,以不同浓度GNPs溶液孵育48 h后的神经元作为实验组?采用TEM观察GNPs在细胞内的分布;采用CCK-8试剂盒检测细胞活力;采用TUNEL法检测细胞凋亡;采用丙二醛(MDA)?超氧化物歧化酶(SOD)试剂盒检测氧化应激水平?结果:TEM及UV-Vis显示硼氢化钠还原法制备的GNPs呈尺寸均一的球形,在溶液中均匀分散?原代培养的大鼠皮质神经元纯度为(82.0 ± 2.3)%?GNPs主要以聚合体形式分布于胞浆?溶酶体?囊泡中?随着GNPs浓度的增高,细胞活性逐渐降低,尤其1 200和2 400 μg/L处理组细胞活力较对照组显著降低(P < 0.05)?TUNEL染色结果显示,随着GNPs浓度的增高,凋亡细胞数逐渐增加,1 200和2 400 μg/L处理组细胞凋亡明显,具有显著性差异(P < 0.05)?随着GNPs浓度的增高,细胞内MDA含量逐渐增加,SOD含量逐渐减少,1 200和2 400 μg/L处理组与对照组相比MDA含量显著升高,SOD含量显著降低(P < 0.05)?结论:5 nm GNPs能降低大鼠皮质神经元的细胞活力,促进其凋亡,其机制可能与氧化应激损伤相关?     

Glu-GNPs对人肺腺癌细胞株A549放射增敏的初步探讨

目的探讨偶联葡萄糖的纳米金颗粒(Glu-GNPs)对人A549细胞的放射增敏作用及其机制。方法噻唑蓝(MTT)法检测低浓度(≤20 nmol/L)Glu-GNPs联合放射对A549细胞存活的影响,克隆形成检测Glu-GNPs对A549细胞的放射增敏作用,流式细胞仪(FCM)检测细胞周期及其凋亡。结果低浓度Glu-GNPs对A549细胞生长无明显抑制,联合X射线后具有抑制作用,在15 nmol/L范围内,随浓度增大,抑制作用增强;15 nmol/LGlu-GNPs对A549细胞有放射增敏作用,由Dq、Do计算放射增敏比(SER)分别为1.93、1.10;Glu-GNPs、单纯放射均可诱导细胞凋亡,凋亡率分别为(7.64±1.43)%、(13.46±1.99)%,联合放射组凋亡率为(21.43±1.04)%,显著高于前两组(P<0.01);Glu-GNPs作用后,细胞周期发生变化,表现为S期减少,G2/M期增加(P<0.05)。结论 Glu-GNPs对人肺腺癌细胞株A549具有放射增敏作用,其机制可能为抑制细胞亚致死损伤修复,阻滞细胞于G2/M期,并诱导细胞凋亡。     

关于银纳米颗粒增强脑胶质瘤细胞放射敏感性的研究

目的：通过制备3组尺寸的银纳米颗粒来研究其增强脑胶质瘤放射治疗的效果。方法：采用柠檬酸钠还原体系制备20、50和100nm尺寸的银颗粒,通过扫描电镜来表征;检测3种尺寸的银纳米颗粒的细胞毒性以及确定保证细胞活性的安全剂量;通过MTT比色法和集落形成实验检测3组尺寸的银纳米颗粒对脑胶质瘤细胞放射治疗的效果;通过流式细胞术检测银纳米颗粒对细胞周期以及细胞放疗后凋亡率的影响。结果：20nm的银颗粒能够显著增强脑胶质瘤细胞的放射敏感性,50nm的银颗粒次之,而100nm的银颗粒无明显效应。结论：小粒径的银纳米颗粒能够增强脑胶质瘤细胞的放射敏感性,为临床放射治疗提供新的前景。     

U0126 在 A549 肺腺癌细胞凋亡中的作用

目的:探讨 ERK 信号通路抑制剂 U0126 与 A549 肺腺癌细胞凋亡的关系。 方法:选用不同浓度的 ERK 信号通路抑制剂 U0126(10滋MU0126、20滋MU0126 和 40滋MU0126)作用于 A549 细胞,采用流式细胞术和 TUNEL 原位末端标记法检测细胞的凋亡情况。 结果:U0126 阻断 ERK 信号通路后,细胞凋亡率明显增加,各实 验组明显高于对照组(P<0. 01),但20滋M U0126 组与40滋M U0126 组间无明显差异(P>0郾 05)。 结论:ERK 信号 传导通路与 A549 肺腺癌细胞的凋亡有关,但 A549 肺腺癌细胞凋亡是受多因素的调控。     

不同厂家二甲基亚砜对人肺腺癌A549细胞生长的影响

目的观察不同厂家二甲基亚砜(DMSO)对体外培养的人肺腺癌A549细胞的最小毒性体积分数,为体外筛选抗肺肿瘤药物实验溶媒的选取提供依据。方法将DMSO以不同体积稀释到DMEM完全培养液中,孵育细胞后,显微镜观察细胞形态,采用四甲基偶氮唑蓝显色(MTT)法和流式细胞仪技术(FCM)法分别检测细胞相对活力、细胞凋亡及细胞周期。结果 Sigma公司DMSO体积分数≤0.1%时,肺腺癌A549细胞能够正常生长并保持形态完整,细胞活力与对照组比较差异无统计意义(P0.05),细胞未发生凋亡;当体积分数达到4.0%时,细胞生长异常,细胞相对活力显著降低(P0.01),细胞凋亡率梯度性增高,G1+S期细胞比例增加。国产DMSO对A549细胞的毒性较进口试剂弱,体积分数≤0.5%时,对A549细胞影响较小(P0.05)。结论大剂量DMSO对A549细胞生长毒性明显,可抑制细胞增殖、诱导细胞凋亡以及影响细胞周期;Sigma公司、天津科密欧公司产品DMSO体积分数分别低于0.1%,0.5%时可作为人肺腺癌A549细胞体外筛选抗肿瘤药实验的安全溶媒。     

3-溴丙酮酸联合顺铂抑制肺癌细胞株A549的生长

目的 探讨3-溴丙酮酸(3-bromopyruvate,3-BrPA)联合顺铂体外抗肺癌A549细胞的作用及其可能机制.方法 采用CCK-8检测不同浓度的3-BrPA、顺铂单用及联用对A549细胞的增殖抑制;选择低于半数抑制浓度(IC50)的40 μmol/L 3-BrPA与1 mg/L顺铂单独和联合作用A549细胞48 h后,用倒置相差显微镜观察细胞形态变化,流式细胞术检测细胞凋亡;不同浓度的3-BrPA作用A549细胞48 h后,流式细胞术检测细胞周期变化,己糖激酶(hexokinase,HK)活性检测试剂盒检测3-BrPA对细胞内HK活性的影响.结果 CCK-8结果显示,与阴性对照组比较,3-BrPA浓度大于20 μmol/L时对A549细胞有明显抑制作用(P＜0.01),与单用顺铂组比较,3-BrPA联合顺铂组可显著增强顺铂对A549细胞的毒性作用(P＜0.01);3-BrPA作用A549细胞48 h后,倒置显微镜观察到3-BrPA组大多数细胞出现凋亡,而联合用药组细胞凋亡更加明显,部分区域出现细胞碎片等坏死征象;流式细胞术结果显示,3-BrPA和顺铂单用组及联合用药组细胞凋亡率均明显高于阴性对照组(P＜0.01),且联合用药组细胞凋亡率明显高于单独用药组(P＜0.01);细胞周期检测结果显示3-BrPA可将A549细胞阻滞于G1期;HK活性结果显示:与阴性对照组比较,3-BrPA组HK活性明显降低(P＜0.01).结论 3-BrPA有抑制肺癌A549细胞增殖、诱导其凋亡的作用,且与顺铂具有协同抗肺癌A549细胞增殖作用,其机制可能为抑制肺癌细胞糖酵解,进而影响肺癌细胞能量代谢.     

NaHCO_3弱碱性溶液促进A549细胞凋亡机制研究

目的研究NaHCO_3弱碱性溶液对A549细胞凋亡的影响,探讨其凋亡发生机制。方法实验分为对照组和NaHCO_3弱碱性溶液处理实验组,分别培养24h和48h,流式细胞检测仪检测肺腺癌A549细胞生长的细胞周期和细胞凋亡情况。结果与对照组相比,实验组肺腺癌A549细胞生长周期出现明显停滞现象,在细胞周期G2期比较集中,进一步说明NaHCO_3弱碱性溶液可以诱导A549细胞凋亡。结论 NaHCO_3弱碱性溶液诱导A549细胞出现凋亡,其具体机制与改变了肿瘤细胞生长的酸性微环境相关。     

昆明山海棠总生物碱诱导肺腺癌A549细胞凋亡与细胞周期改变

目的 为探讨昆明山海棠总生物碱(alkaloidsfrom Tripterygium Hypoglaucum Hutch,THHa)抗肺癌机制,研究THHa诱导肺腺癌A549细胞凋亡与细胞周期改变的关系.方法 应用形态学观察、DNA凝胶电泳、流式细胞仪检测分析肺腺癌A549细胞凋亡及细胞周期改变.结果 THHa能够诱导肺腺癌A549细胞凋亡并主要作用于细胞周期S期(S期细胞增多).结论 THHa可能通过诱导细胞凋亡而抑制A549细胞增殖,这种作用与细胞周期有关,这些有助于将来肺癌治疗方案的合理设计.     

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…