SEA-McAb融合蛋白对SMMC-7221细胞的抑制作用及机制

目的 研究葡萄球菌肠毒素A(SEA)与单克隆抗体(McAb)融蛋白的抗肝癌细胞活性及其机制.方法 制备健康人外周血单个核细胞(PBMC)作为效应细胞.MTT法测定SEA-McAb融合蛋白对人肝癌细胞SMMC-7221的体外抑制率;采用荧光抗体流式细胞术测定肿瘤细胞HLA-I和HLA-DR抗原分子的表达;酶联免疫吸附(ELISA)法检测上清液中细胞因子IL-2、IL-12、TNF-α和IFN-γ的水平.结果 肿瘤细胞在上清波作用后,HLA-I和HLA-DR的表达均有明显上升(P<0.01).分泌IL-2、IL-12、TNF-α和IFN-γ均较对照组显著增加(P<0.01).SMMC-7221明显被抑制(P<0.01).结论 SEA-McAb通过激发PBMC的杀伤作用,对肝癌细胞株表现出较强的靶向抑制作用.     

索拉菲尼联合细胞因子活化的杀伤细胞对肝癌的杀伤效应研究

目的 探讨索拉菲尼联合细胞因子活化的杀伤细胞(cytokine-induced killer,CIK)对肝癌细胞的体外杀伤效应.方法 取健康人外周血单个核细胞,经IFN-γ、IL-1、IL-2和抗CD_3单抗诱导CIK细胞成熟,流式细胞仪检测培养第1、7、14天的CIK细胞表型.Cell Counting Kit-8试剂盒(CCK-8)检测索拉菲尼、CIK细胞及两者联合对肝癌细胞BEL-7402的体外杀伤效应,并用Annexin V-FITC细胞凋亡检测试剂盒检测肝癌细胞的凋亡率.结果 CIK细胞经多种细胞因子诱导培养后,CD_3+CD_(56)~+细胞比例明显上升,14d时CD_3~+CD_(56)~+ 细胞比例达(46.83±2.53)%,与第1天比差异有显著性(P<0.01),细胞量扩增了近1 000倍.索拉菲尼和CIK细胞在适当浓度及效靶比联合应用时表现出杀伤活性增强(P<0.01).索拉菲尼联合CIK细胞诱导的肝癌细胞BEL-7402凋亡率达(70.56±2.58)%,明显高于索拉菲尼组(45.38±1.76)%和CIK组(52.18±8.63)%(P<0.05).结论 索拉菲尼与细胞因子活化杀伤细胞疗法联合应用能明显增强对肝癌细胞的体外杀伤效应.     

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.     

rIL-2、TNF-α、IFN-γ与抗CD3-抗胶质瘤双特异性抗体的协同细胞毒作用

目的:观察细胞因子rIL-2、TNF-α、IFN-γ对抗CD3-抗胶质瘤双特异性抗体(双抗)的协同作用,进一步提高双抗对人脑胶质瘤的治疗作用.方法:以人脑胶质瘤细胞株SHG-44为靶细胞,健康人或胶质瘤患者的外周血单个核细胞(PBMC)为效应细胞,以18 h 3H-TdR掺入释放法测定细胞毒活性,采用单比实验和联合作用等对比分析方法,分析各细胞因子(rIL-2、TNF-α、IFN-γ)对双抗诱导的细胞毒性作用的影响.结果:rIL-2、TNF-α、IFN-γ均可协同提高双抗对人脑胶质瘤的细胞毒作用(P<0.05).来源于恶性胶质瘤患者的效应细胞活性较正常人低,rIL-2与双抗可协同增强其活性.结论:细胞因子rIL-2、TNF-α、IFN-γ可协同提高双抗对效应细胞的活化,充分激活效应细胞,增强抗肿瘤免疫能力.     

IFN-γ、IL-12联合CD80基因转染对肝癌细胞免疫效应的影响

目的观察IFN-γ、IL-12对转染CD80基因前后肝癌细胞表面分子CD80的表达,评价CD80分子联合细胞因子对肝癌细胞杀伤效应的影响。方法取HepG2细胞和经逆转录病毒载体介导CD80转染的HepG2/CD80细胞,分别经IFN-γ、IL-12处理后,流式细胞仪检测CDS0分子的表达水平。LDH释放试验检测刺激前后肝癌瘤苗对CTL杀伤效应的影响。结果HepG2细胞表面CD80分子表达水平低,转染后HepG2/CD80细胞的表达增高。经IL-12、IFN-γ细胞因子诱导后,HepG2/CD80细胞表面的CD80基因表达进一步增强。细胞因子联合CD80基因转染提高细胞毒性淋巴细胞对肝癌细胞的杀伤作用,与对照组比较有显著差异(P<0.05)。结论(1)IL-12、IFN-γ刺激转染CD80肝癌细胞共刺激信号的进一步表达。(2)CD80基因转染联合IL-12、IFN-γ明显增强CTL对肝癌细胞的杀伤作用,提示两者存在协同作用。     

慢性HCV感染患者PBMC体外Th1类细胞因子分泌及其相关因素分析

目的:研究慢性HCV(丙型肝炎病毒)感染患者外周血单个核细胞(PBMC)体外Th1类细胞因子的分泌,进一步了解HCV感染慢性化和肝损伤机制. 方法:体外培养慢性HCV感染患者PBMC 72 h后,ELISA检测培养上清Th1类细胞因子(IL-2,IFN-γ,TNF-α),荧光定量PCR检测患者血清HCV RNA含量,RT-PCR检测HCV RNA亚型. 结果:与正常人相比,IFN-γ在HCV RNA阴性患者中明显增高,而在RNA阳性患者中无明显升高. TNF-α则不论RNA滴度的高低均较正常人显著增高. RNA阳性患者ALT(丙氨酸氨基转移酶),AST(门冬氨酸氨基转移酶)水平明显高于阴性患者. 不同HCV基因型感染的患者IFN-γ,TNF-α的分泌无显著差异. 结论:IFN-γ在清除病毒中起重要作用,TNF-α是引起肝脏损伤的重要因素之一.     

CpG ODN活化外周血单个核细胞对淋巴瘤HUT-78细胞杀伤作用的研究

目的 探讨CpG寡聚脱氧核苷酸(CpG ODN)在肿瘤免疫治疗中的作用.方法 将CpG ODN刺激体外培养的人外周血单个核细胞(PBMC),ELISA法检测其分泌细胞因子白细胞介素-12(IL-12)、干扰素-γ(INF-γ)的表达, MTT法检测CpG ODN作用后的PBMC对淋巴瘤细胞株HUT-78细胞杀伤活性.结果 与对照组相比,经CpG ODN作用后的PBMC对HUT-78细胞杀伤活性显著提高(P＜0.05),同时CpG ODN能明显促进PBMC分泌细胞因子IL-12、INF-γ(P＜0.05).结论 CpG ODN可增强PBMC介导的抗淋巴瘤细胞的作用,其杀伤活性与分泌细胞因子有关. Abstract： Objective To study the immunotherapy role of CpG ODN(CpGoligodeoxynucleotide) on tumor.Methods After treated with synthetic CpG ODN for 48 hours, the inhibiting effect of PBMC on HUT-78 cells were detected by MTT method.The secretions of TNF-γ and IL-12 from PBMC determined using ELISA method.Results The PBMC treated with CpG ODN enhanced their antitumor effect on HUT-78 cells and increased the secretion levels of TNF-γ and IL-12.Conclusions CpG ODN increases the cytotoxicity of PBMC on HUT-78 cells in vitro, as well as facilitates the IL-12 and TNF-γ secretion.     

Effect of chemotherapy on cytolytic capacity and cytokine production of CD8 + T cells from NHL patients with chronic HBV infection

目的:探讨合并慢性HBV感染的非霍奇金淋巴瘤(non-hodgkin's lymphoma,NHL)患者接受化疗后其CD8 +T细胞的杀伤能力及分泌细胞因子能力是否有影响.方法:选择合并HBV慢性感染并且HLA-A2阳性的NHL患者.从其外周血分离纯化CD8 +T细胞,经PHA刺激后与人肝癌细胞株HepG2.2.15共培养,检测CD8+T对肝癌细胞的杀伤及分泌TNF-α、INF-γ的能力.结果:CD8 +T细胞的杀伤能力及分泌TNF-α、INF-γ的水平呈现出效:靶比依赖性,而化疗对杀伤能力和分泌TNF-α、INF-γ的能力没有影响.结论:慢性HBV感染的NHL患者接受化疗不影响其CD8+T细胞的杀伤能力及分泌TNF-α、INF-γ的能力.     

Vγ9Vδ2 T细胞联合顺铂增强对鼻咽癌细胞的杀伤作用

目的探讨Vγ9Vδ2 T免疫细胞联合顺铂（CDDP）对鼻咽癌细胞的杀伤作用。方法采用CCK8法观察低浓度CDDP、唑来膦酸（ZOL）联合Vγ9Vδ2 T细胞对鼻咽癌细胞增殖活性的影响;流式细胞术检测Vγ9Vδ2 T细胞肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）和CD107a的分泌及鼻咽癌细胞表面MICA和MICB的表达;Western blotting检测MICA和MICB的蛋白表达。结果随着CDDP浓度的增加,鼻咽癌CNE2Z细胞存活率降低,CDDP联合Vγ9Vδ2 T细胞的杀伤作用优于单独使用CDDP对CNE2Z细胞的杀伤作用（P < 0.05）。ZOL对Vγ9Vδ2 T细胞的增殖有促进作用,随着ZOL浓度提高,ZOL联合Vγ9Vδ2 T细胞对CNE2Z细胞的杀伤作用增强（P < 0.05）。低浓度CDDP和ZOL联合Vγ9Vδ2 T细胞可增强对鼻咽癌细胞的杀伤作用,当效靶比为10:1时,联合处理的杀伤效果最好,差异有统计学意义（P < 0.05）。低浓度CDDP、ZOL处理的鼻咽癌细胞可增强Vγ9Vδ2 T细胞免疫因子CD107a、TNF-α和IFN-γ的分泌（P < 0.05）。低浓度CDDP联合Vγ9Vδ2 T细胞明显上调鼻咽癌细胞表面MICA和MICB的表达（P < 0.01）。结论CDDP能增强Vγ9Vδ2 T细胞对鼻咽癌细胞的杀伤作用,其机制可能与促进鼻咽癌细胞表面MICA和MICB的表达及增加Vγ9Vδ2 T细胞TNF-α和IFN-γ的分泌有关。     

芦荟多糖联合Anti-CD3 McAb、rhIL-2干预的PBMC对HepG2生长的影响

目的探讨芦荟多糖(AP)的抗肿瘤作用机制。方法采用AP单独及联合Anti-CD3McAbr、hIL-2诱导干预外周血单个核细胞(PBMC)的方法建立效应细胞;采用MTT法测定效应细胞及其上清液对靶细胞HepG2生长的影响;ELISA法测定干预后各组PBMC分泌IFN-γ、TNF-α的水平,测定效应细胞及其上清液对靶细胞上清液中IFN-γ、TNF-α和AKP水平的影响。结果AP单独及其联合Anti-CD3McAbr、hIL-2可提高PBMC分泌IFN-γ水平,对TNF-α水平没有明显影响;效应细胞的上清液对HepG2的生长无明显抑制作用,而效应细胞细胞悬液对HepG2的生长有抑制作用,可升高混合培养后上清液中TNF-α、IFN-γ水平,降低AKP水平。结论AP体外抗肿瘤作用与提高免疫和促进肿瘤坏死因子释放有关。     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.