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In this series of 65 cases of aplastic anemia, 26 cases were treated by the kidney-tonifying and mediating method, 19 cases by western drugs, and the remaining 20 cases only by tonifying the kidney as controls. The results showed that the kidney-tonifying and mediating method was significantly superior in the total effective rate to the method of western drugs and that of tonifying the kidney alone (P<0.01), and it was also superior in the improvement of symptoms, myelogram, and peripheral blood count to the other two methods.
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供者特征对rhG-CSF动员的外周血和骨髓混合移植物中免疫细胞组成的影响
近,我们采用GIAC(即G,粒细胞集落刺激因子+I,强的免疫抑制+A,抗胸腺细胞球蛋白+C,外周血与骨髓联合)技术体系成功跨越了HLA不合屏障,从根本上解决了异基因造血干细胞移植(Allo-HSCT)供者来源缺乏的问题[1-2].大量资料显示,供者特征影响移植物的细胞组成,后者又与Allo-HSCT的预后密切相关[2-3].我们从供者的年龄、性别、体重等方面研究供者特征对粒细胞集落刺激因子(G-CSF)动员的外周血和骨髓混合移植物中免疫细胞组成的影响.
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临床病例讨论(step by step)——自体干细胞移植后发热、呼吸困难
患者女性,28岁,确诊非霍奇金淋巴瘤(弥漫大B细胞型Ⅳ期A IPI 2分)18月.已行10程化疗(R-CHOP×6次,R-CHO×1次,R-DICE×1次,R-MINE×2次),1程外周血造血干细胞动员,复查PET/CT提示病情复发,又予3程巩固化疗(R-ES-HAP×2次,DICE×1次)后,拟行自体外周血造血干细胞移植(autologous peripheral blood stem cell transplantation,APBSCT)于2010-03-24入院.
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血管免疫母细胞性T细胞淋巴瘤骨髓/外周血瘤细胞免疫表型分析
血管免疫母细胞性T细胞淋巴瘤(AITL)是一类以异形T淋巴细胞增生伴显著血管增生,以及滤泡树突状细胞增生为主要表现,为一类来源于成熟T细胞的淋巴瘤[1].过去AITL瘤细胞形态学诊断缺乏特异性的鉴别特征,而近来AITL瘤细胞CD10[2]和CXCL13表达意义的发现为AITL的诊断提供了有力的支持,为了更深入了解AITL瘤细胞的免疫表型特征及发病机制,我们利用流式细胞术分析AITL患者骨髓及外周血有核细胞表面CD分子标志,期望获得更多信息.
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外周血中血管内皮前体细胞的分离和鉴定
新血管的形成主要有两种方式,一是血管形成(vasculogenesis),即造血血管母细胞(hemangioblast)原位分化成内皮细胞并形成原始毛细血管丛的过程[1,2];二是血管新生(angiogenesis),指由已存在的血管以出芽的方式形成新血管的过程[3,4].一直以来,人们认为血管形成只发生于胚胎发育时期,在出生后并不存在.1997年,Asahara等[5]发现在人的外周血中存在血管内皮前体细胞(endothelial progenitor cells, EPC),该种细胞可参与出生后的多种病理和生理过程中的新血管形成[6],此现象同胚胎时期的血管形成相类似,故称作出生后血管形成(postnatal vasculogenesis).我们采用外周血作为材料来源,建立了一种稳定可重复的分离、鉴定末梢血液EPC的方法,为进一步研究EPC在生理和病理过程中的作用提供重要的实验基础.
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维吾尔族宫颈癌患者外周血中CD+4CD+25CDlow/-127调节性T淋巴细胞与细胞因子的临床意义
宫颈癌是位居妇科肿瘤第一的恶性疾病[1].如何有效预防和治疗宫颈癌,提高患者生存质量是目前的研究热点.CD+4 CD+25 Treg在维持自身耐受和免疫稳定的同时,也抑制着免疫系统对肿瘤的免疫应答[2].经TGF-β和IL-10处理的细胞可获得明显的抑制效应性T淋巴细胞增殖的作用,炎症因子IFN-γ则与Treg起相反的作用[3].维吾尔族是宫颈癌高发民族,死亡率高[4].本研究通过流式细胞术检测维吾尔族宫颈癌患者外周血CD+4 CD+25 CDlow/-127 Treg细胞含量,采用ELISA检测血清中细胞因子TGF-β、IL-10及IFN-γ的水平,以便有针对性地开展新疆地区宫颈癌防治工作.
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AIM To study the detection of micrometastasis in peripheral blood of patients with gastric carcinomas andits clinical significance.METHODS A cytokeratin 19 (CK19)-specific nested reverse transcriptase-polimerase chain reaction (RT-PCR) assay was developed to detect CK19 expressing cancer cells, the sensitivity was determined by serialdilution method using CK19 expressing gastric cancer cells, the specificity was assessed by examining 12negative controls and 12 positive controls. Then pre-operative peripheral blood from 42 patients with gastriccancer was detected and the relationship between positive results and biological behavior was studied.RESULTS CK19mRNA was expressed in all the 12 gastric cancer tissues but not in peripheral blood from12 healthy individuals;sensitivity of nested RT-PCR amplification for CK19mRNA was confirmed to be 1/106 by serial dilution method using human gastric cancer line SGC-7901; micrometastases in pre-operativeperipheral blood were detected in 13 (30,9%) patients with gastric carcinomas, the frequency ofmicrometastasis in peripheral blood was significantly correlated with tumor size,depth of invasion and TNMstage (x2 test, P<0.05).CONCLUSION Nested RT-PCR amplification for CK19mRNA is a sensitive and specific method for thedetection of micrometastases in peripheral blood in gastric cancer patients; pre-operative detection ofmicrometastasis in peripheral blood may be helpful in the prediction of tumor progression.
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Objective:In order to explore the radioprotective effects of the expression of hematopoietic growth factors regulated by radio-inducible promoter on radiation injury. Methods:The human FL (Flt3 ligand) cDNA and EGFP (enhanced green fluorescent protein) cDNA were linked together with IRES and then inserted into the eukaryotic expression vector pCI-Egr, which was constructed by substituting CMV promoter in pCIneo with the Egr-1 promoter (Egr-EF). The vector was transferred into human bone marrow stromal cell line HFCL by lipofectin. The transduced cell clones (HFCL/EF) had been selected by the addition of G418. The cells were exposed to γ-radiation by 60 Co source for 0.5-20Gy. The expressions of transduced cells were detected with FACS, Northern blot ELISA and CFU assay. The HFCL/EF and CD34+ cells from human umbilical cord blood were one after the other transplanted i.v. into sublethally irradiated severe combined immunodeficient (SCID) mice. The white blood cell amount in peripheral blood and human cell engrafted in recipent mice were detected by flow cytometry and CFU-GM etc. Results:The activity of EGFP in transduced cells increased by 3.1 fold as compared to non-transduced cells at 18h after exposure to 2.5Gy. The amounts of secreted FL in serum-free supernatants of Egr-EF increased by 605.46±107.21pg/ml, which were significantly higher than the control group (214.45±35.61pg/ml). The effects of FL in HFCL/EF cultural supernatants on expansion of CD34+ cells derived from cord blood in the presence of SCF, IL-6 and IL-3 were also studied. The results showed that at day 10 of culture the number of CD34+ cells increased by 173. 09±11.58×103/ml, which was significantly higher than that of non-radiation group(68. 04± 13. 73 × 103/ml). It showed that radiation can enhance the ability of the supernatants containing FL of HFCL/EF to expand early hematopoietic progenitor cells and protect hematopoietic cells from radiation-injury effects. The HFCL/EF and CD34+cells from human umbilical cord blood were one after the other transplanted i. v. into sublethally irradiated severe combined immunodeficient (SCID) mice. In contrast to two control groups (HFCL and HFCL/F), HFCL/EF (the Egr-1 regulatory element-drived expression of FL gene therapy) resulted in a proportionally obvious increase in the number of the white blood cell at early stage after radiation, while no significant differences were found for CD45+ 、CD34+ cells in bone marrow cells. In contrast to two control groups (HFCL and HFCL/F), HFCL/EF (the Egr1 regulatory element-drived expression of FL gene therapy) resulted in a proportionally obvious increase in the number of the white blood cell at early stage after radiation, without significant differences being found for CD45+、CD34+ 、CFU-GM and marrow nucleared cells in bone marrow cells. Conclusions:The results suggested both in vivo and in vitro use of the gene therapy of FL gene regulated by Egr-1 promoter could protect hematopoiesis from irradiation-induced damage.
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自体外周血造血干细胞移植在儿童神经母细胞瘤中的应用
神经母细胞瘤(Nueroblastoma,NB )是儿童常见的外周神经来源的恶性肿瘤,约占儿童恶性肿瘤的7%~10%[1]。根据INSS分期标准NB临床分期分为1~4期以及特殊4S期,4期恶性程度高、预后差。Mauricio A 等[2]2006年报道153例 IV期NB长期生存率仅为34%。目前骨髓移植或自体外周血干细胞移植(autologous peripheral blood stem cell transplantation,APBSCT)对于治疗儿童Ⅳ期NB具有一定的临床疗效[3]。APBSCT是经大剂量放化疗预处理,清除受体体内的肿瘤细胞及异常克隆细胞,阻断发病机制,然后移植入自体造血干细胞,使其重建正常造血免疫,而达到治疗目的的一种治疗手段。So Young Yoo 等[4]2013年报道90例高危NB患儿中因复发、进展、死亡等因素73例按计划行APBSCT治疗,15例移植后复发或进展,7例死亡,58例获随访,总生存率为65%(38/58),总无事件生存42.2%(38/90),因此APBSCT治疗高危NB具有积极的意义。虽然20世纪80年代末以来, APBSCT已逐渐为人们所接受及熟悉,但对于低质体重儿的移植安全、C D34是否分选纯化、骨髓净化、二次移植的意义等仍是目前临床关注的热点。本文将就上述问题作初步的阐述。
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神经母细胞瘤患者骨髓和外周血微量病灶监测方法的评估
神经母细胞瘤(neuroblastoma, NB)是儿童常见的颅外实体瘤.NB患者诊断时骨髓(bone marrow, BM)有否转移是一个重要的预后因素.对于接受化疗后自身外周血干细胞移植(peripheral blood stem cell transplant, PBSCT)的患者,其移植物污染可能导致远期复发.本研究运用三色荧光流式细胞术(flow cytometry, FCM)检测CD-45/CD+56/CD+81表达的NB细胞以检测BM和外周血的微量病灶.报告如下.
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哮喘豚鼠模型肺泡灌洗液与外周血嗜酸性粒细胞凋亡率及存活率的研究
2002年5月-2003年12月,我们在哮喘豚鼠模型中比较支气管肺泡灌洗液(BALF)和外周血两种不同来源的嗜酸性粒细胞(EOS)的凋亡及在体外存活情况,报告如下.
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再生障碍性贫血外周血Th1/Th2、Tc1/Tc2异常的临床意义
多数获得性再生障碍性贫血(简称再障)是T细胞介导的自身免疫性疾病.儿童再障强化免疫抑制治疗(intensive immunosuppressive therapy,IIST)有效率达71.8%[1],但至今未有明确的疗效预测指标.
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系膜增生型原发性肾病综合征患儿细胞毒性T淋巴细胞相关抗原4基因多态性、蛋白表达及外周血淋巴细胞凋亡的研究
系膜增生型(mesangial proliferative glomerulonephritis,MsPGN)原发性肾病综合征(idiopathic nephrotic children,INS)(INS-MsPGN)是我国儿童INS常见的病理类型之一,临床多表现为糖皮质激素(glucocorticoid,GC)不敏感或耐药,是难治性INS中常见的一种病理类型,其发病机制和GC耐药机制至今尚不清楚.
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自体外周血干细胞移植治疗下肢缺血性疾病的研究进展
下肢缺血性疾病是一种常见病及多发病,包括下肢动脉硬化闭塞症,下肢血栓闭塞性脉管炎和雷诺氏病等疾病,治疗手段包括药物、血管旁路移植术和介入手术等,治疗方法无实质性改进,疗效也不理想[1],往往导致患者肢体坏死而被迫行截肢手术,严重影响患者的身心健康.自体外周血干细胞移植(Autologous peripheral blood stem cell transplantation,APBSCT)是近年来发展迅速的一项治疗技术,实验和临床应用证明了APBSCT是治疗下肢缺血性疾病的一种很有应用前景的方法.
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矽肺宁对早期煤工尘肺患者外周血T淋巴细胞亚群的影响
为研究中药矽肺宁对煤工尘肺患者外周血T淋巴细胞亚群(SPBL)的影响,我们测定了30例煤工尘肺患者口服矽肺宁前后外周血SPBL的分布情况,报道如下.
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环氧乙烷致外周血淋巴细胞微核率及染色体畸变率异常增高一例
患者男,42岁,从事环氧乙烷处理工作9年,既往健康.2001年1月工作时,发生管道破裂,环氧乙烷气体喷出,而后出现头痛、头晕、乏力、心慌、胸闷、手颤、失眠、健忘,并伴有尿频、尿痛等症状,3个月后入院治疗.实验室检查:尿常规、尿细菌培养及血生化各项指标均正常,其中WBC 3.6×109/L,Hb 120 g/L;外周血淋巴细胞微核出现率36‰(正常范围0~3‰),其制备方法采用甲基纤维素法,每份标本在油镜下计数1000个淋巴细胞中所含微核的细胞数,结果以千分率(‰)表示;染色体的制备方法按《人类染色体方法手册》[1]制片,畸变类型按WHO统一标准分类,即每份标本选择细胞完整、图像清晰、分散度好的中期相,在油镜下计数100个中期细胞,以百分率(%)表示.患者的染色体畸变数目如下:染色体断裂10个,染色单体断裂2个,染色体断片2个,双着丝粒4个,核内复制1个,4N∶1,47∶1,48∶1,畸变率总计为22%(正常值0~0.3%).
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长期低水平苯接触对工人外周血调节性T细胞的影响
高浓度苯对血液系统和免疫系统具有明显的毒性作用[1-3].但长期低水平苯接触对工人免疫系统的影响还不清楚.有研究表明,职业苯接触工人CD8淋巴细胞亚型比例异常增多,而CD8淋巴细胞可能参与了骨髓造血细胞的免疫损伤.
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保存全血中泛素对人外周血单个核细胞分泌细胞因子的影响
同种异体输血产生的免疫调节不良作用,称为输血相关免疫调节(Transfusion-Related Immunomodulation,TRIM)[1].我们通过体外观察保存全血血浆中泛素对LPS刺激的人外周血单个核细胞(PBMNC)分泌细胞因子的影响,探讨其在TRIM中的作用.
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自体外周血干细胞移植治疗下肢缺血疾病的规范化护理
自体外周血干细胞移植治疗下肢缺血性疾病方法操作简单、创伤小、安全有效,干细胞取自本人,不存在排异反应[1].我院利用自体干细胞移植术治疗下肢缺血性疾病日趋成熟,临床治疗效果明显.但病人在治疗过程中主动、及时、有效的护理配合对整个移植过程起到了关键作用,现总结如下.