全反式维甲酸抑制IL-23/IL-17通路促进小鼠移植皮肤存活的研究

【摘要】目的探讨全反式维甲酸(ATRA)灌胃对小鼠同种异基因皮肤移植存活时间的影响及白细胞介素(IL)-23、IL-17通路在其中的作用。方法以DBA/2小鼠为供者,Babl/c小鼠为受者建立皮肤移植模型。随机将受者分为对照组、小剂量组和大剂量组,分别在术前1d至术后14d或判定皮肤死亡之日每天给3组小鼠分别灌胃注射玉米油、10mg/kg和30mg/kg ATRA玉米油溶液。术后观察各组皮肤移植物存活时间,皮肤组织切片检测病理改变,酶联免疫吸附试验检测血清IL-23和IL-17水平,实时荧光定量PCR检测移植皮肤中IL-23、转录因子孤核受体（ROR）γt和IL-17mRNA表达。结果与对照组比较,小剂量组和大剂量组皮肤移植存活时间延长（P<0.05）,炎症细胞浸润及组织破坏程度轻,血清IL-23水平降低（P<0.05）,而两治疗组间差异无统计学意义。对照组、小剂量组及大剂量组血清IL-17水平依次降低（P<0.05）。小剂量组和大剂量组皮肤移植物中IL-23、RORγt和IL-17mRNA表达水平均较对照组低（P<0.05）,而两治疗组间差异无统计学意义。结论ATRA灌胃可显著延长小鼠移植皮肤存活时间,其机制可能与抑制IL-23、RORγt、IL-17mRNA表达和蛋白分泌有关。     

国产注射用特立帕肽的过敏性及免疫原性研究

目的 考察国产注射用特立帕肽在动物中的过敏性及免疫原性.方法 对豚鼠和家兔分别于0d和14 d注射一定浓度的特立帕肽,观察豚鼠全身变态反应及对家兔的免疫原性,并采用ELISA分析家兔血清抗体的变化规律.结果 给药后豚鼠未见明显全身变态反应.家兔免疫原性实验结果表明,低剂量组（1 μg/kg）血清中的IgE含量经免疫后无明显上升;中剂量组（5μg/kg）和高剂量组（10 μg/kg）血清中IgE含量在首次免疫后略有快速上升,但随后中剂量组恢复正常,而高剂量组在加强免疫后IgE含量仍有一定上升.结论 国产注射用特立帕肽在拟定的人用剂量范围内具有很好的安全性,可供皮下注射给药.     

PCI术后血清IL-8/IL-10比值变化及相关因素分析

目的:探讨经皮冠状动脉介入治疗(PCI)术后白细胞介素(IL)-8/IL-10比值的变化及相关影响因素。方法:行冠状动脉造影(CAG)患者103例,其中稳定型心绞痛单纯造影组(SA1组)31例、稳定型心绞痛PCI组(SA2组)34例、不稳定型心绞痛PCI组(UA组)38例。检测各组患者PCI术前及术后24h血清高敏C-反应蛋白(hs-CRP)、IL-8及IL-10水平。结果:(1)SA2组及UA组术后IL-8/IL-10比值高于术前(P<0.05或P<0.01)。(2)术后UA组IL-8/IL-10比值均高于SA1及SA2组(P<0.05)。(3)术前IL-8/IL-10比值及不稳定型心绞痛状态是术后IL-8/IL-10比值的影响因素。结论:PCI术后存在促炎/抗炎细胞因子失衡,机体炎症反应增强,且以术前存在细胞因子失衡状态及不稳定型心绞痛患者更明显。     

环孢素A调节自噬对人滋养细胞氧化应激损伤的影响

目的探讨环孢素A(CsA)调节自噬对人滋养细胞氧化应激损伤的保护作用。方法将细胞分为对照组、氧化应激组、低剂量CsA(2μmol/L)组和高剂量CsA(4μmol/L)组。流式细胞仪检测HTR-8/SVneo滋养细胞凋亡,硫代巴比妥酸比色法检测丙二醛(MDA),化学比色法检测超氧化物歧化酶(SOD),化学荧光法检测活性氧(ROS),Western blot检测蛋白表达。结果对照组、氧化应激组、低剂量CsA组和高剂量CsA组HTR-8/SVneo细胞凋亡率分别为4.24%±0.72%、38.15%±4.63%、27.24%±3.16%、18.45%±2.28%,各组间比较差异有统计学意义(P<0.01)。CsA组细胞MDA和ROS显著低于氧化应激组(P<0.01),且高剂量CsA组低于低剂量CsA组(P<0.01);CsA组细胞SOD显著高于氧化应激组(P<0.01),且高剂量CsA组高于低剂量CsA组(P<0.01)。对照组、氧化应激组、低剂量CsA组和高剂量CsA组HTR-8/SVneo细胞自噬蛋白LC3Ⅱ表达的相对灰度值分别为0.23±0.05、0.11±0.02、0.38±0.07、0.75±0.13,各组间比较差异有统计学意义(P<0.01)。结论CsA可以增强滋养细胞自噬,对氧化应激损伤诱发的滋养细胞凋亡起到抑制作用。     

细辛木脂素抗心脏移植急性排斥反应的实验观察

目的:观察细辛木脂素(Herba Asari Ligini,HAL)对大鼠同种异位心脏移植急性排斥反应的治疗效果,为抗排斥药物的研究提供实验依据。方法:建立大鼠同种异位心脏移植模型,受体自术前1天至术后14天分别给予 CsA10mg/kg/d、CsA2.5mg/kg/d、HAL50mg/kg/d、HAL50MG/kg/d+CsA2.5mg/kg/d 观察移植心存活时间。结果:单用 HAL 组的移植心存活时间比对照组长(P<0.01),但短于 CsA10mg/kg/d 组(P<0.01);CsA2.5mg/kg/d 组与 HAL50mg/kg/d合用组的移植心存活时间与 CsA10mg/kg/d 组相拟(P>0.05)。结论:HAL 具有抗排斥作用,但效果低于治疗量(10mg/kg/d)的 CsA;HAL 与 CsA 抗急性排斥反应具有协同作用。     

贝伐单抗对胃移植癌裸鼠VEFG、IL-8、bFGF表达的影响

目的 探讨贝伐单抗对胃移植癌裸鼠血管内皮生长因子（VEGF）、白细胞介素-8（IL-8）、碱性成纤维细胞生长因子\r\n（bFGF）表达的影响。方法 选择BALB/c 裸鼠为研究对象,于颈背部皮下注射胃癌细胞株以建立胃移植癌裸鼠模型,随机分为空白对照组、低剂量贝伐单抗组和高剂量贝伐单抗组。低剂量组注射11 mg/kg贝伐单抗,高剂量组注射22 mg/kg贝伐单抗,空白对照组注射等体积生理盐水,比较三组裸鼠的肿瘤生长情况、血清以及肿瘤组织VEFG、IL-8、bFGF 含量。结果 治疗7 d、14 d、21 d 后,贝伐单抗组肿瘤体积明显小于空白组,且高剂量贝伐单抗组明显小于低剂量贝伐单抗组（P<0.05）。治疗21d 后,贝伐单抗组的肿瘤质量明显低于空白组,且高剂量贝伐单抗组明显低于低剂量贝伐单抗组（t=2.189～4.807,P<0.05）;贝伐单抗组裸鼠血清与肿瘤组织中VEGF、IL-8、bFGF 含量均明显低于空白组,且高剂量贝伐单抗组血清与肿瘤组织中VEGF、IL-8、bFGF 含量明显低于低剂量贝伐单抗组（t=2.443～7.745,P<0.05）。结论 贝伐单抗对胃移植癌裸鼠模型的肿瘤生长具有抑制作用,同时能够降低VEGF、IL-8、bFGF 的表达。     

不同剂量瑞舒伐他汀治疗急性脑梗死的临床疗效及其对血清炎症因子水平的影响观察

目的 探讨急性脑梗死患者采用不同剂量的瑞舒伐他汀治疗,对患者血清炎症因子和疗效的影响。方法 选取急性脑梗死患者200例,随机分为低剂量组和高剂量组,各100例。低剂量组采用10 mg/d剂量的瑞舒伐他汀治疗,高剂量组采用20 mg/d剂量的瑞舒伐他汀治疗。观察两组患者治疗效果、血清炎症因子水平情况、脑神经功能、生活质量以及不良反应情况。结果 高剂量组治疗总有效率高于低剂量组(P<0.05)。治疗后,高剂量组IL-1β、IL-6以及TNF-α水平明显低于低剂量组(P<0.05);高剂量组NIHSS评分低于对照组,ADL评分高于对照组(P<0.05)。两组不良反应发生率对比差异无统计学意义(P>0.05)。结论 大剂量瑞舒伐他汀治疗急性脑梗死患者效果较好,能明显降低患者炎症因子水平,提高生活质量,改善脑神经功能,且安全性较高,值得临床应用及推广。     

肺癌手术期间血清IL-10含量及临床意义

目的测定肺癌患者手术期间血清白细胞介素-10(interleukin-10,IL-10)含量,探讨肺癌手术期间血清IL-10含量的变化及临床意义。方法用放射免疫吸附法测定了39例肺癌患者手术期间及16例肺良性肿瘤患者和30例正常人血清IL-10含量并进行对照比较。结果肺癌组术前IL-10含量明显高于肺良性肿瘤组及正常人组(P<0.01),根治切除的肺癌患者血清IL-10含量术后明显下降,与术前相比,有非常显著性差异(P<0.01);开胸探查的肺癌患者术后24h轻度升高,72h恢复到术前含量。结论动态观察血清IL-10含量对肺癌诊断、判断手术效果、预测病情、指导术后治疗有重要的临床意义。     

老年胸腰椎骨折60例术后炎症反应与疼痛关系分析

目的 探讨老年胸腰椎骨折患者术后炎症反应与疼痛的关系，为患者术后的无痛化管理及快速康复提供参考依据。方法 对我科收治的老年胸腰椎骨折患者60例，观察并记录术后12、24、48 h患者疼痛VAS评分(据疼痛程度分组：轻度1～3分，中度4～6分，重度7～10分),检测术前，术后12、24、48 h血清CRP、TNF-α、IL、INF-γ水平，且进行比较分析。结果 术后12 h疼痛程度：轻度24人、中度35人、重度18人，中、重度组血清IL-6、IL-10、TNF-α、CRP水平均高于轻度组(P<0.05),重度组血清IL-6、CRP水平高于中度组(P<0.05);术后24 h疼痛程度：轻度35人、中度17人、重度8人，中、重度组血清IL-6、IL-10、TNF-α、CRP水平均高于轻度组(P<0.05),中、重度组各炎性因子水平无明显差异(P>0.05);术后48 h疼痛程度：轻度47人、中度8人、重度5人，中、重度组血清IL-6、IL-10、CRP水平均高于轻度组(P<0.05),重度组IL-6及CRP水平高于中度(P<0.05),而TNF-α在各组间...     

右美托咪啶超前镇痛对腹腔镜下胆囊切除术患者应激反应与细胞免疫功能的影响

目的探讨右美托咪啶超前镇痛对腹腔镜下胆囊切除术(LC)患者应激反应与细胞免疫功能的影响。方法择期行LC患者90例,年龄27～66岁,体重45～84kg,ASAⅠ～Ⅱ级,随机分为3组,每组30例,对照组(C组)不予右美托咪啶;不同剂量右美托咪啶组,分别于麻醉诱导前静脉注射右美托咪啶0.4μg.kg-1(D1组)和0.8μg.kg-1(D2组)。观察3组患者术后4、8、12、24、48h的视觉模拟评分(VAS)及术前、术后24h、2d、3d血清皮质醇及白细胞介素-2(IL-2)溶度。结果 VAS评分:术后4、8、12、24、48h的VAS评分低于C组,差异有统计学意义(P<0.01),D2组的VAS评分低于D1组,差异有统计学意义(P<0.05);3组患者术前血清皮质醇及IL-2溶度差异无统计学意义(P>0.05);术后24h、2d、3d血清皮质醇含量均较术前高,差异有统计学意义(P<0.01或P<0.05),D1、D2组均低于C组,差异有统计学意义(P<0.01);D2组术后24h、2d、3d血清IL-2含量与术前比较无显著性差异(P>0.05);而C、D1组术后24h、2d、3d血清IL-2含量与术前比较有降低,差异有统计学意义(P<0.01或P<0.05)。结论右美托咪啶超前镇痛有利于降低LC患者术后应激反应的水平,而且对患者术后细胞免疫功能有一定保护作用。     

IL-10基因克隆化与原核重组IL-10蛋白制备的实验研究

目的克隆化人IL-10基因及重组蛋白的制备为IL-10基因表达在小儿炎症性、过敏性疾病中的作用及治疗等提供有力的证据。方法采用过敏症患者发作时期的外周血,提取总RNA。以总RNA为模板,特异性引物进行反转录,RT-PCR进行IL-10基因筛选。PCR产物经酶切分析和DNA序列测定后,构建重组质粒pTrchHis2B-hIL10原核高效表达载体,通过大肠杆菌(工程菌)JM109经IPTG诱导表达重组hIL-10蛋白,并经与Ni-NTA树脂中的6×His配体结合进行亲和层析纯化,表达产物进行SDS-PAGE电泳分析。结果DNA测序显示克隆化人IL-10全长开放读框准确,表达读码框正确。SDS-PAGE分析显示,原核表达系统表达的重组蛋白量和纯度满意。结论采用以总RNA为模板的RT-PCR一步法,能简便、可靠地获得hIL-10基因产物,所构建的原核表达载体pTrcHis2B-hIL10,能够快速、高效制备rhIL-10蛋白,为细胞因子hIL-10功能的进一步研究及临床应用奠定了基础。     

鼻咽癌中白细胞介素-10、白细胞介素-12检测及其意义

目的 检测鼻咽癌患者白细胞介素-10(interleukin-10,IL-10)及白细胞介素-12(interleukin-12,IL-12)浓度.方法 用酶联免疫法(Enzyme Linked Immunosorbent Assay,ELISA)检测43例鼻咽癌患者血清IL-10及IL-12浓度,并用22例健康成年人血清作为对照.结果 鼻咽癌组IL-10浓度明显高于对照组,鼻咽癌组IL-12浓度明显低于对照组,IL-10浓度在晚期T分期组(T3/T4)明显高于早期T分期组(Ⅰ/Ⅱ)(P<0.01)、晚期临床分期组(Ⅲ/Ⅳ)明显高于早期临床分期组(Ⅰ/Ⅱ),但在不同性别、年龄组及有无淋巴结转移组浓度差异无统计学意义(P>0.05).而IL-12在晚期T分期组(T3/T4)明显低于早期T分期组(Ⅰ/Ⅱ)(P<0.01)、晚期临床分期组(Ⅲ/Ⅳ)明显低于早期临床分期组(Ⅰ/Ⅱ),但在不同性别、年龄组及有无淋巴结转移组浓度差异无统计学意义(P>0.05).且IL-10及IL-12浓度呈负相关关系.结论 鼻咽癌中高浓度的IL-10可能抑制IL-12的产生,帮助肿瘤细胞免疫逃避,损害机体肿瘤免疫,从而促进肿瘤发展.     

Association of IL-10 level and IL-10 promoter SNPs with specific antibodies in penicillin-allergic patients

Objective Our aim was to investigate the hypothesis that the sera interleukin-10 (IL-10) level and polymorphic nucleotides within the IL-10 gene promoters would link to specific IgE and IgG production and the expression of penicillin allergy. Methods One hundred and two patients and 86 healthy subjects were chosen for assay of serum IL-10 level by enzyme-linked immunosorbent assay (ELISA) and type −1082 G/A and −819 C/T alleles by sequence-specific primer polymerase chain reaction (SSP-PCR). Radioallergosorbent test (RAST) and ELISA were used to examine eight types of specific immunoglobulin-E (IgE) and IgG antibodies, respectively, which included four types of antibodies to major and minor antigenic determinants. Results Compared with control subjects and patients with negative-specific IgE, there were significantly lower levels of IL-10 in patients with positive-specific IgE (P < 0.05). Similarly, there were significantly lower levels of IL-10 in patients with positive-specific IgG compared with normal controls and allergic patients with negative-specific IgG (P < 0.05). The visible negative correlations existed between IL-10 and four types of specific IgE [benzylpenicilloyl (BPO), phenoxomethylpenicilloyl (PVO), benzylpenicillanyl (BPA), amoxicillanyl (AXA)], and patients with three or more positive-specific IgE had significantly lower IL-10 levels than normal controls (P < 0.01). There was a declining trend for IL-10 level in serum with the increase in types of positive-specific IgE. But there was no significant difference in serum IL-10 level between the positive skin-test group and the allergic-history group. Compared with controls and patients with negative antibodies, a significantly decreased frequency of the −1082 G allele was present in patients with positive antibodies (P < 0.01). The allele T and TT genotype at −819 C/T position had lower frequency in the negative-specific IgG group than that in the positive group and controls (P < 0.01). Conclusions Positive specific IgE and IgG are associated with decreased IL-10 level in allergic reaction to penicillins. The distributions of genotype and frequency of allele at the −1082 G/A position may be associated with the production of both specific IgE and IgG antibodies.     

IL-10抑制脂多糖诱导的Hela细胞IL-15和IL-6转录

目的研究IL-10对脂多糖(LPS)诱导的Hela细胞IL-15mRNA和IL-6 mRNA表达的影响,分析其激活的信号转导通路。方法培养的Hela细胞,经不同浓度的LPS及IL-10单独或联合处理后,提取细胞总RNA和总蛋白,RT-PCR分析IL-15和IL-6转录水平的变化,Westernblot分析信号转导通路蛋白变化。结果RT-PCR分析得知①1ng～10μgLPS刺激Hela细胞12h后,IL-15 mRNA和IL-6 mRNA水平均明显上调(与对照组比较:P<0.01),且存在剂量依赖关系,100ng/mL时达峰值;100ng/mLLPS刺激Hela细胞0～24h,IL-15 mRNA和IL-6 mRNA水平亦明显上调(与对照组比较:P<0.01),24h内存在时间依赖关系,12h时达峰值。②单独IL-10(10ng/mL)作用于Hela细胞12h后,IL-15 mRNA和IL-6 mRNA没有明显变化(与对照组比较:P>0.05)。不同浓度的IL-10(1,10,100ng/mL)均下调100ng/mLLPS诱导的Hela细胞IL-15 mRNA和IL-6 mRNA的表达,且浓度越高IL-10抑制作用越明显。Western blot分析显示LPS主要通过磷酸化信号蛋白PI3K/AKT和ERK1/2上调IL-15和IL-6转录,IL-10能阻断AKT的磷酸化而对ERK1/2的磷酸化没有影响。结论IL-10可抑制LPS诱导的炎症细胞因子IL-15和IL-6的转录,这可能与其阻断AKT的磷酸化有关,因而,IL-10可能应用于某些临床感染性疾病的预防和治疗。     

Effect of PTU on IL-12 and IL-10 in psoriasis

Propylthiouracil (PTU), an antithyroid thioureylene with immunomodulatory properties, has been shown to be effective in the therapy of patients with plaque psoriasis. The mechanism of action of antithyroid thioureylenes in psoriasis remains unknown. Propylthiouracil is a commonly used agent in the treatment of patients with Graves' hyperthyroidism, a condition associated with elevated levels of interleukin-12 (IL-12), which fall significantly after propylthiouracil treatment. IL-12 is believed to play a pivotal role in the development of psoriasis. Production of IL-12 is modulated by the anti-inflammatory cytokine IL-10. The effect of PTU on IL-12 and IL-10 levels was, therefore, studied in twelve patients with plaque psoriasis. Treatment with 300 mg of PTU daily in divided doses for three months produced significant improvement of the PASI and histological scores in the patients. Serum IL-12 concentrations were undetectable at baseline and did not change with treatment. IL-10 concentrations were 1.39 +/- 1.49 pg/ml (mean +/- SD) at baseline, and showed no significant change after 2 weeks (1.63 +/- 1.61 pg/ml and 12 weeks 1.15 +/- 1.58 pg/ml of treatment with PTU. The data suggest that the clinical improvement with patients with psoriasis treated with PTU is not due to a fall in circulating IL-12 or a rise in IL-10 concentrations. Although the drug may have effects on lesional production of these cytokines this is not reflected in the circulating levels. It is speculated that the beneficial effect is likely mediated by an inhibitory effect on keratinocyte proliferation or promotion of apoptosis in these proliferated keratinocytes.     

骨性关节炎患者血清及组织中白细胞介素-10测定

目的探讨白细胞介素-10（IL-10）在骨性关节炎发病机制中的作用。方法采用ELISA法对16例膝关节骨性关节炎（OA）患者的血清、关节液、关节软骨、滑膜及5例类风湿性关节炎（RA）患者的血清、关节液进行了IL-10测定。结果OA患者关节软骨及滑膜中的IL-10含量明显高于血清及关节液,RA患者关节液中IL-10含量明显高于血清中。结论软骨及滑膜中IL-10的高表达说明其在OA的关节损伤中可能起双向调节作用。     

MDA-7/IL-24 is a unique cytokine--tumor suppressor in the IL-10 family

The melanoma differentiation associated gene-7 (mda-7) cDNA was isolated by virtue of being induced during melanoma differentiation. Initial gene transfer studies convincingly demonstrated potent antitumor effects of mda-7. Further studies showed that the mechanism of antitumor activity was due to induction of apoptosis. Most striking was the tumor-selective killing by mda-7 gene transfer--normal cells were unaffected by Adenoviral delivery of mda-7 (Ad-mda7). A variety of molecules implicated in apoptosis and intracellular signaling are regulated by Ad-mda7 transduction. Different apoptosis effector proteins are regulated in different tumor types, suggesting that Ad-mda7 may regulate various signaling pathways. mda-7 encodes a secreted protein, MDA-7, which has now been designated as IL-24, and is a novel member of the IL-10 cytokine family. MDA-7/IL-24 protein is actively secreted from cells after mda-7 gene transfer. In human peripheral blood mononuclear cells (PBMC), STAT3 activation by MDA-7/IL-24 is followed by elaboration of secondary Th1 cytokines, demonstrating that MDA-7/IL-24 is a pro-Th1 cytokine. Furthermore, MDA-7/IL-24 is antagonized by the prototypic Th2 cytokine IL-10. MDA-7/IL-24 protein is endogenously expressed in cultured NK and B-cells and is also expressed in dendritic cells in tissues. MDA-7/IL-24 protein is expressed in nevi and melanoma primary tumors, to varying degrees, but is rarely expressed in malignant melanoma or other human tumors evaluated. Indeed, loss of MDA-7/IL-24 protein expression correlates strongly with melanoma tumor invasion and disease progression. The "bystander" effects proposed for MDA-7/IL-24 protein include immune stimulation, antiangiogenesis and receptor-mediated cytotoxicity. Thus, mda-7 is a unique multifunctional cytokine in the IL-10 family and may have potent antitumor utility in a clinical setting.     

慢性心力衰竭与白细胞介素-17及白细胞介素-10的相关性研究

目的 探讨慢性心力衰竭与白细胞介素(IL)-17、IL-10的相关性.方法 采用酶联免疫吸附试验(ELISA)检测37例慢性心力衰竭患者(慢性心衰组)和34例健康体检者(对照组)血浆IL-17和IL-10水平,并进行相关性分析.结果 慢性心衰组血浆IL-17水平高于对照组[(46.37±10.57) ng/L vs(32.45±4.55)ng/L],IL-10水平低于对照组[(27.49±4.19) ng/L vs(32.71±4.38) ng/L],差异均有统计学意义(t=7.09,5.46,P＜0.01);慢性心力衰竭患者血浆IL-17与IL-10呈负相关(r=-0.63,P＜0.01).结论 慢性心力衰竭的发病可能与体内IL-17、IL-10的水平失衡存在相关.     

肝细胞肝癌患者血清IL-18、IL-10变化的分析

目的探讨血清IL-18和IL-10在肝细胞肝癌(HCC)患者中的变化及其意义。方法采用ELISA法检测65例HCC患者(HCC组)血清IL-18和IL-10水平,并与正常对照组比较。分析HCC患者合并肝硬化(LC)与合并慢性乙型肝炎(CHB)间血清IL-18、IL-10水平的差异。结果HCC组血清IL-18、IL-10水平明显均高于正常对照组(P<0.05);HCC合并LC患者血清IL-18水平明显高于合并CHB患者(P<0.05),而两者间血清IL-10无统计学差异(P>0.05)。结论 HCC患者体内以Th2型免疫反应为主,其血清IL-18、IL-10的水平同时升高可以抑制机体的免疫功能。     

不同浓度HAES在正常人体外对IL-10和IL-12的影响

目的 探讨不同浓度的羟乙基淀粉类血浆代用品(贺斯6%HAES200/0.5)对正常人体免疫系统白介素-10(IL-10)和白介素-12(IL-12)的影响,以检验其是否抑制正常人体的免疫系统.方法 正常志愿者30名,严格无菌条件下抽取静脉血各20 ml,并以肝素抗凝,在超净台内分离出单个核细胞,计数后将细胞平均分成5组...