舌癌患者CIK细胞的体外培养及其免疫活性

目的研究舌癌患者外周血CIK细胞的增殖能力、细胞表型和细胞毒作用,寻求对舌癌过继免疫治疗的新途径。方法提取20例舌癌患者和20例健康人外周血单核细胞用IFN-γ、抗CD3mAb、IL-2和IL-1β共同培养诱导CIK细胞,流式细胞仪检测细胞表型特征,用MTT法检测细胞对Tca8113的杀伤活性;同时培养舌癌患者LAK细胞、CD3AK细胞和舌癌患者CIK细胞在增殖和杀瘤活性上作比较。结果舌癌患者20ml外周血经过一个周期培养平均获得的CIK细胞数低于健康对照组(P<0.05),但舌癌患者CIK细胞经过周期培养也较培养前的数量明显增高,于12~21d达高峰。舌癌患者CIK细胞与LAK相比,增殖倍数和杀伤活性均明显提高(P<0.05);舌癌患者CIK增殖倍数与CD3AK相比,早期差异无显著性,但后期增殖倍数及杀伤活性有显著提高(P<0.05)。结论舌癌患者CIK细胞体外增殖倍数和杀瘤活性明显优于LAK和CD3AK细胞,低于正常人CIK的增殖能力,而舌癌患者CIK细胞杀伤活性与正常人来源CIK细胞差异无显著性(P>0.05),有望作为自体过继免疫治疗舌癌的一种新方法。     

CD3AK、LAK与CIK细胞生物学特性的研究

①目的探讨CD3AK、LAK及CIK细胞的诱导、增殖能力及杀伤活性的变化.②方法采用CD3单抗和IL-2刺激诱生扩增CD3AK细胞,用IL-2刺激诱生LAK细胞,采用第0天加入IFN-γ,第1天加入IL-1,CD3单抗,IL-2诱生CIK细胞,观察它们的扩增情况;用MTT法以K562和H7402细胞为靶细胞,测定CD3AK、LAK及CIK细胞的杀伤活性.③结果CD3AK和CIK细胞的扩增能力远大于LAK细胞(P＜0.05).CIK细胞和CD3AK相比,在前期无明显差异,至第19、22天时CIK细胞的扩增能力显著高于CD3AK的扩增倍数(P＜0.05).CD3AK和CIK细胞对K562和H7402细胞的杀伤活性均显著高于LAK细胞(P＜0.05).④结论CIK细胞具有更高的扩增能力和更强的杀伤活性,是肿瘤过继免疫治疗中更为有效的杀瘤效应细胞.     

20例慢性乙型肝炎患者细胞因子活化的杀伤细胞动态观察

目的:动态观察慢性乙型肝炎患者细胞因子活化的杀伤(CIK)细胞的增殖及杀伤活性.方法:抽取10例健康人及20例慢性乙型肝炎患者的外周血,常规分离单个核细胞(PBMC),加IFN-γ,IL-2及抗人CD3单克隆抗体后培养,诱导CIK细胞形成.于培养后3,6,12,24,30 d,取培养的细胞,用流式细胞仪检测细胞表面CD3,CD4,CD8,CD3 CD56及CD95 CD28分子的表达水平、增殖及杀伤活性.结果:慢性乙型肝炎患者CIK细胞的增殖及杀伤活性均低于正常对照组.培养不同时间乙肝患者CIK细胞的增殖倍数、杀伤活性和上述表面标志的表达水平,均较培养前明显增高,于培养后12 d达高峰.结论:慢性乙型肝炎患者CIK细胞的增殖倍数、杀伤活性均低于正常人,但明显高于培养前;这可能是导致HBV感染持续发展的原因之一.     

细胞因子诱导的杀伤细胞体外扩增及其生物学特性的研究

目的:观察细胞因子诱导的杀伤细胞(CIK)的体外增殖能力及其生物学特性。方法:取正常成人新鲜血,经密度梯度离心法,利用连续非贴壁的方法获得单个核细胞(PBMC),加入不同的细胞因子(IFN-γ、IL-1β、IL-2、CD3mAb),诱导生成CIK细胞,以LAK细胞作为对照。观察CIK细胞的扩增情况,用流式细胞仪检测其表面标志,MTT法测定其杀瘤活性。结果:诱导16d后,CIK细胞的增殖率达到高峰,具有较强的扩增能力,经过22d培养,总的细胞数可扩增至100多倍;CIK细胞和LAK细胞相比,在前期无明显差异,至18、21d时扩增能力显著高于LAK细胞的扩增倍数(P<0.01)。CIK细胞是一群异质细胞群,CD3 CD56 细胞为其主要的效应细胞,经过22d培养显著增加,为(55.15±5.49)%,其绝对值可增加1500多倍。CIK对A-549细胞的杀伤活性均显著高于LAK细胞(P<0.05),具有较强的杀瘤能力。结论:CIK细胞具有较强的扩增和杀瘤能力,是肿瘤过继免疫治疗中更为有效的杀瘤效应细胞。     

细胞因子诱导的杀伤细胞体外增及其生物学特性的研究

目的观察细胞因子诱导的杀伤细胞(CIK)的体外增殖能力及其生物学特性.方法取正常成人新鲜血,经密度梯度离心法,利用连续非贴壁的方法获得单个核细胞(PBMC),加入不同的细胞因子(IFN-γ、IL-1β、IL-2、CD3mAb),诱导生成CIK细胞,以LAK细胞作为对照.观察CIK细胞的扩增情况,用流式细胞仪检测其表面标志,MTT法测定其杀瘤活性.结果诱导16 d后,CIK细胞的增殖率达到高峰,具有较强的扩增能力,经过22 d培养,总的细胞数可扩增至100多倍;CIK细胞和LAK细胞相比,在前期无明显差异,至18、21 d时扩增能力显著高于LAK细胞的扩增倍数(P＜0.01).CIK细胞是一群异质细胞群,CD3 CD56 细胞为其主要的效应细胞,经过22 d培养显著增加,为(55.15 5.49)%,其绝对值可增加1500多倍.CIK对A-549细胞的杀伤活性均显著高于LAK细胞(P＜0.05),具有较强的杀瘤能力.结论CIK细胞具有较强的扩增和杀瘤能力,是肿瘤过继免疫治疗中更为有效的杀瘤效应细胞.     

鼻咽癌患者自体细胞因子诱导的杀伤细胞体外增殖及杀瘤活性

目的 研究鼻咽癌患者外周血细胞因子诱导的杀伤细胞（CIK）的增殖能力、对鼻咽癌细胞株HONE1的细胞毒作用,寻求鼻咽癌过继免疫治疗的新途径。方法 提取24例鼻咽患者外周血单个核细胞,用IFN-γ、抗CD3mAb、IL-2和IL-1β共同培养诱导CIK细胞,计数不同时间点的细胞数,用MTT法检测细胞对鼻咽癌细胞株HONE1的杀伤活性,用ELISA法测定CIK细胞分泌种细胞因子的水平,同时培养CD3AK细胞和LAK细胞,与CIK细胞在增殖和杀瘤活性上进行比较。资料用SPSS13.0软件进行分析,P＜0.05为统计有差异性。结果 鼻咽癌患者CIK细胞表现出很强的体外增值活性及杀瘤活性,与CD3AK细胞和LAK细胞相比,其增殖活性和对鼻咽癌细胞株HONE1的杀伤活性均明显提高（P＜0.05）,且与正常人自体CIK细胞相比无明显差别（P＞0.1）。结论 鼻咽癌患者CIK细胞体外增殖能力和杀瘤活性明显优于CD3AK和LAK细胞,为鼻咽癌患者的临床过继免疫治疗提供了一种新的有效地治疗手段。     

正常人和肿瘤患者CIK细胞的体外增殖能力及抗肿瘤作用的对比分析

目的:探讨正常人和肿瘤患者CIK细胞体外增殖能力及抗肿瘤作用的差异。方法:分离正常人和肿瘤患者的外周血单个核细胞(PBMC),加入细胞因子(CK),体外诱导CIK细胞,用细胞计数板法观察CIK细胞增殖能力及随时间变化的趋势,用MTT法检测其对不同肿瘤细胞株的杀伤活性。结果:与正常人CIK细胞相比,肿瘤患者CIK细胞的增殖速度较慢,其最大增殖倍数低(P<0.01);CIK细胞的增殖速度与年龄有关,即低年龄段的CIK细胞增殖能力强;正常人的CIK细胞对乳腺癌细胞株(MDA-MB-231)、肺巨细胞癌细胞株(BE-1)和前列腺癌细胞株(PC-3)的杀伤活性明显高于肿瘤患者的CIK细胞(P<0.01);不同年龄段肿瘤患者的CIK细胞对肿瘤细胞的杀伤率差异有统计学意义(P<0.05)。结论:CIK细胞的体外增殖力及对肿瘤细胞的杀伤活性与年龄有关,正常人CIK细胞的体外增殖力及对肿瘤细胞的杀伤活性高于肿瘤患者的CIK细胞。     

流式细胞术检测CIK的免疫表型和体外杀伤活性

目的：探讨细胞因子诱导的杀伤细胞（cytokine-induced killer,CIK）的免疫表型和体外杀伤活性。 方法:从健康人外周血分离淋巴细胞,经过IFNγ、CD3McAb、IL-2诱导并培养,获得大量的CIK。采用流式细胞术检测CIK的表型;以CFSE标记靶细胞来区分效应细胞,再以ＰＩ染色,用FACS检测靶细胞的死亡率。 结果:CIK于第22天达到增殖高峰,并高度表达CD3、CD11a、CD54和HLA-DR;中度表达CD3/CD56、CD25、CD28、CD69和FasL;CD16表达不增加。CIK对肿瘤细胞K562、 HL-60、Hela、SMMC7721和A375均表现出杀伤活性,其中对K562、HL-60、Hela 3种细胞的杀伤作用较强;而对SMMC7721、A375的作用不明显。 结论:细胞因子IFNγ、CD3McAb和IL-2体外诱导的单个核细胞具有强大的增殖力和杀伤活性。     

细胞因子诱导的杀伤细胞对人胃癌高侵袭腹膜转移细胞株体内外杀瘤过程中多种细胞因子的影响

目的:观察细胞因子诱导的杀伤细胞(CIK细胞)自分泌细胞因子及其体内外杀瘤过程中多种细胞因子含量的变化,探讨CIK细胞对抗肿瘤的作用机制.方法:取正常人的外周肝素抗凝血分离外周血单个核细胞(PBMC),体外诱导成CIK细胞;同时建立人胃癌高侵袭转移瘤(OCUM-2MD3)细胞裸鼠腹膜移植模型.检测CIK细胞中白介素-2(IL-2)、肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)等细胞因子的分泌及其对OCUM-2MD3细胞株体内外杀瘤时上述细胞因子的变化.结果:①CIK细胞培养过程中其上清液IL-2、TNF-α、INF-γ、GM-CSF含量随时间增长而升高,至2周左右达最高峰,随后下降;②体外培养CIK细胞以E:T=25:1对OCUM-2MD3肿瘤细胞作用后,IL-2含量减低明显(P<0.05),TNF-α、GM-CSF含量均降低,INF-γ含量升高;③体内抗瘤实验结果表明,CIK治疗组血清中IL-2、TNF-α、IFN-γ、GM-CSF的含量均较生理盐水对照组显著增高(P<0.05).结论:CIK细胞分泌的细胞因子在2周左右具有最大生物学活性;CIK细胞可能通过其自身分泌IL-2、TNF-α、IFN-γ、GM-CSF等细胞因子参与抗肿瘤作用.     

IL-24增强细胞因子诱导的杀伤细胞对HepG2的细胞毒活性

目的 寻求增强CIK细胞毒活性的有效方法,以满足临床过继免疫治疗的需要.方法 从健康人外周血中提取出单个核细胞,第1天加入IFN-γ,第2天加入IL-1、CD3mAb、IL-2诱导CIK细胞;另一组与IL-1、CD3mAb、IL-2同时加入IL-24.细胞计数法测定细胞的增殖,MTT法测定细胞杀伤活性,比较两者有无差别.透射电镜下观察CIK细胞对肿瘤细胞的杀伤和肿瘤细胞的改变.结果 未加IL-24培养组增殖率高于加IL-24培养组,两者比较有明显差异(P＜0.05).加IL-24培养组细胞各个效靶比杀伤活性均达到90%以上,明显高于其它各组.透射电镜下可见加IL-24培养组凋亡和坏死的肿瘤细胞比未加IL-24培养组明显增多.结论 CIK细胞诱导过程中加入IL-24能明显增强它的杀伤活性.     

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…