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。     

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.     

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。     

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.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

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…     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.