自体外周血CD_(34)~ 造血干细胞移植治疗多发性骨髓瘤

[目的]探讨自体外周血CD 造血干细胞移植治疗多发性骨髓瘤的可行性。[方法]应用CliniMACS系统分选纯化患者外周血CD 造血干细胞 ,用流式细胞仪检测分选纯化结果。选马法兰200mg/m2 作为预处理方案。[结果]分选纯化结果 ,单个核细胞数为3.7136×108,活性率96.6% ,CD 细胞数3.68018×108,即5.52×106/kg。移植后患者骨髓像获得完全缓解。移植相关并发症少且易于控制。[结论]自体外周血CD 造血干细胞移植治疗多发性骨髓瘤是安全、有效、可行的。     

自体外周血CD+3434造血干细胞移植治疗多发性骨髓瘤

[目的]探讨自体外周血 CD+34造血干细胞移植治疗多发性骨髓瘤的可行性。[方法]应用 Clini MACS系统分选纯化 患者外周血 CD+34造血干细胞,用流式细胞仪检测分选纯化 结果。选马法兰 200mg/m2作为预处理方案。[结果]分选纯化结 果,单个核细胞数为 3.7136× 10 8,活性率 96.6％, CD+34细胞数 3.68018× 10 8,即 5.52× 10 6/kg。移植后患者骨髓像获得完全缓 解。移植相关并发症少且易于控制。[结论]自体外周血 CD+34 造血干细胞移植治疗多发性骨髓瘤是安全、有效、可行的 。     

应用C1iniMACS细胞分选系统纯化自体外周血干细胞的探讨

目的 探讨应用CliniMACS细胞分选系统纯化自体外周血干细胞的效果。方法 4例非霍奇金淋巴瘤患者,经Vp-16+G-CSF动员后,采集外周血中造血干细胞,并用CliniMACS细胞分选系统作干细胞纯化分选,3例患者作2次采集后行1次分选,1例患者经3次采集后作2次分选。通过单个核细胞计数及流式细胞术测定CD34+细胞,分析纯化效果。结果 4例患者进行7例次干细胞采集,循环量6000～8 000ml,采集前体内CD34+细胞数为每千克体重(12.77±11.21)×106,经采集得到CD34+细胞数为每千克体重(4.70±4.17)×106,CD34+细胞采集效率为(40.91±33.35)％,对5例次磁珠分选结果,标记前CD34+细胞数为每千克体重(6.07±3.18)×106,分选后获得的CD34+细胞数为每千克体重(4.03±2.28)×106,CD34+细胞回收率为(64.19±13.35)％。终产物细胞悬液量为40～45 ml,CD34+细胞纯度为(83.18±6.76)％。结论 应用CliniMACS细胞分选系统获得纯化的自体造血干细胞,可应用于支持大剂量化疗治疗实体肿瘤。     

纯化的自体CD34~+细胞移植治疗恶性血液病临床对照观察

 目的　分析CD34+ 纯化自体造血干细胞移植治疗恶性血液病的临床疗效。方法　 11例病人进行了CD34+ 纯化自体造血干细胞移植 ,8例患者进行了未分选的自体造血干细胞移植 ,动员方案为CTX+Vp 16联合G CSF。CD34+ 细胞分选采用CliniMACS系统。结果　分选后CD34+ 细胞纯度达到(88.99± 6 .88) % ,回收率达到 (6 9.0 6± 17.0 7) %。 11例分选患者感染率为 (5 / 11例 ) ,复发率为 (1/ 11例 ) ,死亡率为 (3/ 11例 )。外周血中性粒细胞 >0 .5× 10 9/L时间为 (11± 1)天 ,血小板 >2 0× 10 9/L为(13± 3)天 ,8例未行CD34+ 分选患者的感染率 (4/ 8例 ) ,复发率为 (3/ 8例 ) ,死亡率为 (5 / 8例 )。外周血中性粒细胞 >0 .5× 10 9/L时间 (11± 1)天和血小板 >2 0× 10 9/L(14± 4 )天。结论　利用CliniMACs系统体外进行外周血CD34+ 细胞的富集与纯化纯度、回收率均满意 ,自体移植后造血功能重建顺利 ;与对照相比造血重建时间、感染率...     

自体CD34^＋细胞移植治疗晚期肿瘤

目的采用CD34+细胞体外分选技术对晚期肿瘤患者进行自体CD34+细胞移植,以降低自体移植后肿瘤复发率.方法对15例Ⅲ～Ⅵ期肿瘤患者(多发性骨髓瘤11例,乳癌2例,非霍奇金淋巴瘤和髓母细胞瘤各1例)采用Clini MACS临床型细胞富集仪,利用磁性分选技术收集CD34+和CD34-细胞组分,患者于预处理后,输注分选后的CD34+细胞.结果CD34+细胞体外纯化富集可使CD34细胞获得2.0～5.0个对数的去除;回输CD34+细胞中位数为2.4×10 6/kg,CD34+细胞回收率为64%,纯度为98.2%;移植后白细胞恢复至》1.0×10 9/L和血小板》20×10 9/L的天数(中位数)分别为14 d和13 d.患者总体生存率66.7%(10/15),无疾病生存率53.3%(8/15).结论CD34+细胞移植后获得迅速、稳定的造血重建.体外CD34+细胞纯化富集后移植可望提高晚期肿瘤患者自体移植疗效.     

自体CD34~+细胞移植治疗晚期肿瘤

目的　采用CD34+ 细胞体外分选技术对晚期肿瘤患者进行自体CD34+ 细胞移植 ,以降低自体移植后肿瘤复发率。方法　对 15例Ⅲ～Ⅵ期肿瘤患者 (多发性骨髓瘤 11例 ,乳癌 2例 ,非霍奇金淋巴瘤和髓母细胞瘤各 1例 )采用CliniMACS临床型细胞富集仪 ,利用磁性分选技术收集CD34+和CD34-细胞组分 ,患者于预处理后 ,输注分选后的CD34+ 细胞。结果　CD34+ 细胞体外纯化富集可使CD34-细胞获得 2 .0～ 5 .0个对数的去除 ;回输CD34+ 细胞中位数为 2 .4× 10 6/kg,CD34+ 细胞回收率为 6 4 % ,纯度为 98.2 % ;移植后白细胞恢复至 >1.0× 10 9/L和血小板 >2 0× 10 9/L的天数 (中位数 )分别为 14d和 13d。患者总体生存率 6 6 .7% (10 / 15 ) ,无疾病生存率 5 3.3% (8/ 15 )。结论　CD34+ 细胞移植后获得迅速、稳定的造血重建。体外CD34+ 细胞纯化富集后移植可望提高晚期肿瘤患者自体移植疗效。     

外周血造血干细胞的动员、采集和移植后造血重建

目的对13例健康供者及患者外周血造血干细胞的动员和采集效果进行分析。方法健康供者用rhG-CSF动员;患者采用强化疗加rhG-CSF或和rhGM-CSF动员,COBESpectro血细胞分离机全自动造血干细胞程序采集单个核细胞。结果13例移植中8例一次采集成功,1例(即首例)采集4次,余者采集2次。采集后的MNC6×108/kg～12×108/kg,CD3+4细胞3×106/kg～34×106/kg。移植后全部患者造血均获重建,异基因外周血造血干细胞移植一个月后DNA指纹图均提示植活。结论rhG-CSF可作为健康供者的安全有效动员剂,强化疗加rhG-CSF或/和rhGM-CSF是白血病、淋巴瘤自体外周血造血干细胞移植的有效动员方法之一。     

自体外周血造血干细胞移植治疗多发性骨髓瘤16例临床疗效观察

目的:评价自体外周血造血干细胞移植(autologousperipheralbloodstemcelltransplantation,APBSCT)治疗多发性骨髓瘤的临床疗效。方法:16例确诊多发性骨髓瘤患者接受APBSCT,其中2例接受了二次移植,1例接受CD34+细胞筛选后的自体外周血造血干细胞移植。移植后继续常规化疗,13例患者给予α-干扰素维持治疗。结果:APBSCT可延长多发性骨髓瘤患者的无瘤生存率及总生存率,该组患者3年、5年无瘤生存率分别为18.75%±9.75%、0,平均无瘤生存时间为24.8个月。3年、5年总生存率分别为41.25%±12.72%、18.33%±10.77%,平均总生存时间为37.4个月。本组移植患者的CR率高,达76.92%,接近国外报道。而且,移植后造血重建快,移植相关并发症少。结论:APBSCT是治疗多发性骨髓瘤、改善其预后的重要手段。     

化疗联合单一剂量rhG-CSF用于自体外周血造血干细胞移植的临床研究

背景与目的:总结广东省干细胞多中心研究协作组自1999年6月至2001年12月间55例自体外周血造血干细胞移植治疗造血系统恶性疾病的资料,对化疗联合单一剂量rhG-CSF用于自体外周血造血干细胞移植前动员及移植后造血重建的效果进行研究和评价。方法:全部病例(急性髓细胞性白血病28例,急性淋巴细胞性白血病9例,非霍奇金淋巴瘤14例,其他4例)采用化疗+重组粒系集落刺激因子(rhG-CSF,格拉诺赛特)联合动员方案,其中白血病患者主要采用EA方案,恶性淋巴瘤患者主要采用以CTX为主的方案。rhG-CSF用量为250μg/d,WBC升至>4×109/L后,连续1～2天采集PBSC。移植后+3天开始使用rhG-CSF250μg/d,并观察造血重建情况。结果:动员所需的时间即自化疗开始至采集的平均时间为(18.08±3.63)天,rhG-CSF平均应用剂量为4.15μg·(kg·d)-1,应用时间平均7.12天。55例患者平均采集1.38次,采集到的MNC细胞数为(4.09±1.69)×109/kg,CD34+细胞平均值为8.5×106/kg,CFU-GM平均为(6.1±5.8)×105/kg。WBC恢复至>1.0×109/L及中性粒细胞绝对值>0.5×109/L的中位天数分别为10天和10.5天,全部移植患者均获满意的造血重建。结论:我们采用的EA和以CTX为主的化疗联合单一剂量rhG-CSF,是一种安全有效的动员自体外周血造血干细胞的方法,单一剂量rh     

实体瘤患者外周血造血干细胞的动员与保存

目的 :探讨一种适于晚期恶性肿瘤治疗的自体外周血造血干细胞移植方法。方法 :单用rG -CSF(75μg,q12h×2d)作动员剂 ,用血细胞分离机一次性收集单个核细胞(MNC),4℃贮存。观察WBC、MNC计数、细胞活力及细胞分类 ;用流式细胞仪进行细胞表型检测。结果 :小剂量rG -CSF动员后 ,外周血WBC达(18.71±6.54)×109/L。一次可收集MNC(1.87±0.66)×108/kg。CD34、CD14、CD3 及CD22 细胞分别为4.39 %、3.85 %、4.63 %及3.76 %。CD34 细胞平均约为8.2×106/kg。4℃贮存48～56小时MNC存活率为94.3 %。结论 :对实体瘤尚无骨髓转移的患者 ,用此方法可获得足以重建造血功能的干细胞。     

Paraquat and radiation effects on mouse C3HI 10T12 cells

The dipyridilium compound, paraquat, has been used in conjunction with mouse C₃H $\text{10}\text{T}\text{1}\text{2}$ cells to determine if this superoxide (O₂²) generating agent acts to oncogenically transform, chromosomally after or influence cytokinetics or cellular survival. Paraquat alone is a cytotoxic agent and is additionally a weak radiosensitizer. A 0.1 mM 24 hour treatment results in about 30% cell survival and enhances the cell killing effects of ¹³⁷Cs gamma rays by a factor of about 1.2. The drug appears to function lethally by initiating an interphase cell death, and additionally slows the movement of cycling cells through the cell cycle. It is a poor inducer of SCE's and combined effects with radiation are strictly additive. Paraquat oncogenically transforms cells but not in a dose-dependent manner, yet combined treatments with 3 Gy result in transformation frequencies greater than expected for additive effects. Depending on the endpoint examined, which may be related to the degree of nuclear involvement, paraquat either acts additively (SCE's) or with greater than an additive effect (cell survival and oncogenic transformation).     

p16INK4ahypermethylation and p53, p16 and MDM2 protein expression in Esophageal Squamous Cell Carcinoma

Background  

Tumor suppressor genes p53 and p16 ^INK4a and the proto-oncogene MDM2 are considered to be essential G1 cell cycle regulatory genes whose loss of function is associated with ESCC carcinogenesis. We assessed the aberrant methylation of the p16 gene and its impact on p16 ^INK4a protein expression and correlations with p53 and MDM2 protein expressions in patients with ESCC in the Golestan province of northeastern Iran in which ESCC has the highest incidence of cancer, well above the world average.     

食管癌中CD44V5和nm23-H1基因表达及意义

目的探讨CD44V5,nm23-H1的基因产物表达与食管癌临床病理及预后的关系.方法应用免疫组织化学S-P法对85例食管癌标本进行CD44V5和Nm23-H1的基因产物测定,并对其中50例患者术后随访3年.结果①CD44V5在食管癌中的阳性表达率为61.2%,Nm23-H1在食管癌中的阳性表达率为57.6%.②在食管癌中CD44V5的过表达和Nm23-H1的低表达与食管癌的浸润深度、分化程度、淋巴结转移、临床TNM分期及预后相关(P＜0.05),而与食管癌患者的性别、年龄、肿瘤大小、病理类型均无相关性(P＞0.05).③在食管癌中,CD44V5和Nm23-H1的表达呈负相关(r=-0.439,P＜0.01),且CD44V5的过表达伴有Nm23-H1低表达者发生淋巴结转移的可能性大(P＜0.01).结论CD44V5和Nm23-H1在食管癌中的不同表达与食管癌的淋巴结转移及预后显著相关.CD44V5、Nm23-H1的不同表达在食管癌淋巴结转移中可能起协同作用,可为临床预测食管癌淋巴结转移及评估预后提供重要的参考指标.     

CCNG2在甲状腺肿瘤表达的临床意义及过表达对甲状腺癌细胞增殖与凋亡的影响

目的 细胞周期蛋白G2 (CyclinG2, CCNG2 ) 在甲状腺癌中的表达意义及其过量表达对甲状腺癌SW579细胞的生物学影响。方法 采用免疫组织化学方法、Western blot检测63例甲状腺癌组织及距其癌组织边缘5cm以上的镜下未见癌浸润的63例正常组织中蛋白的表达情况。通过慢病毒转染方法建立过量表达CCNG2的甲状腺癌SW579细胞。采用RT-PCR及Western blot 检测转染后SW579细胞内CCNG2基因的表达水平,MTT比色法及流式细胞术(FCM)观察细胞增殖、凋亡变化情况。结果 免疫组织化学结果表明,CCNG2蛋白在正常甲状腺组织及甲状腺癌组织中的表达阳性率分别为96.8%、33.3%,较正常甲状腺组织明显降低(P＜0.05)。Western blot结果表明,CCNG2蛋白在甲状腺癌组的表达相对量较正常甲状腺组织的表达相对量明显降低,二者比较差异具有统计学意义(P＜0.05)。CCNG2蛋白表达与甲状腺癌患者性别、年龄、肿瘤大小无关(P＞0.05);而与甲状腺癌细胞分化程度、淋巴结转移以及肿瘤临床分期有关(P＜0.05)。Kaplan-Meier生存分析提示,CCNG2表达阳性的甲状腺癌患者的5年总生存率明显高于其表达阴性者,二者比较差异具有统计学意义(P＜0.05)。MTT及凋亡结果表明,CCNG2过量表达能明显抑制SW579细胞增殖能力,细胞凋亡数目明显增多(P＜0.05)。结论 CCNG2在甲状腺癌发生发展过程中占重要作用,可作为判断甲状腺癌患者预后的指标。     

Evaluation of NTHL1, NEIL1, NEIL2, MPG, TDG, UNG and SMUG1 genes in familial colorectal cancer predisposition

Background  

The observation that germline mutations in the oxidative DNA damage repair gene MUTYH cause colorectal cancer (CRC) provides strong evidence that dysregulation of the base excision repair (BER) pathway influences disease susceptibility. It is conceivable that germline sequence variation in other BER pathway genes such as NTHL1, NEIL1, NEIL2, MPG, TDG, UNG and SMUG1 also contribute to CRC susceptibility.     

胃癌中p21WAF1/CIP1、细胞周期素D1、p53表达的相关性

目的探讨p21WAF1/CIP1、细胞周期素D1(cyclin D1)、p53在胃癌中表达之间的相关性.方法应用原位杂交技术检测p21WAF1/CIP1 mRNA、细胞周期素D1 mRNA及免疫组化技术检测p53蛋白在胃癌中的表达.结果 p21WAF1/CIP1 mRNA在癌组织及癌旁正常粘膜中阳性表达率各为93.15%(68/73)及76.71%(56/73),二者相比具有显著差异(P<0.05).Cyclin D1 mRNA在癌组织及癌旁正常粘膜中阳性表达率各为54.79%(40/73)及30.16%(22/73),二者具有显著差异(P<0.05).p53蛋白在胃癌中的阳性表达率为32.87%(24/73), p53过表达者,其p21WAF1/CIP1 mRNA表达较p53阴性者为低,二者存在显著差异(P<0.05).p21WAF1/CIP1表达与细胞周期素D1表达呈负相关.结论 p21WAF1/CIP1、Cyclin D1、p53的异常表达及它们之间可能存在的相互作用,对于胃癌的发生发展具有重要意义.     

Polymorphisms in GSTT1, GSTM1, NAT1 and NAT2 genes and bladder cancer risk in men and women

Background  

Cigarette smoking is an established risk factor for bladder cancer. Epidemiological and biological data suggest that genetic polymorphisms in activating and detoxifying enzymes may play a role in determining an individual's susceptibility to bladder cancer in particular when in combination with specific environmental exposures such as cigarette smoking. N- acetyltransferase (NAT) enzymes, NAT1 and NAT2, are involved in the activation and detoxification of tobacco smoke constituents. Polymorphisms in these genes alter the ability of these enzymes to metabolize carcinogens, as certain allelic combinations result in a slow or rapid acetylation phenotype. Glutathione S-transferases (GSTs) also detoxify tobacco smoke constituents, and polymorphisms within the GSTM1 and GSTT1 genes can result in a complete lack of enzyme activity.     

Association of single nucleotide polymorphisms in ATM, GSTP1, SOD2, TGFB1, XPD and XRCC1 with clinical and cellular radiosensitivity

Purpose

To examine the association of polymorphisms in ATM (codon 158), GSTP1 (codon 105), SOD2 (codon 16), TGFB1 (position -509), XPD (codon 751), and XRCC1 (codon 399) with fibrosis and also individual radiosensitivity.

Methods and materials

Retrospective analysis with 69 breast cancer patients treated with breast-conserving radiotherapy; total dose delivered was restricted to vary between 54 and 55 Gy. Fibrosis was evaluated according to LENT/SOMA score. DNA was extracted from blood samples; cellular radiosensitivity was measured using the G0 assay and polymorphisms by PCR-RFLP and MALDI-TOF, respectively.

Results

Twenty-five percent of all patients developed fibrosis of grade 2 or 3. This proportion tends to be higher in patients being polymorphic in TGFB1 or XRCC1 when compared to patients with wildtype genotype, whereas for ATM, GSTP1, SOD2 and XPD the polymorphic genotype appears to be associated with a lower risk of fibrosis. However, none of these associations are significant. In contrast, when a risk score is calculated based on all risk alleles, there was significant association with an increased risk of fibrosis (per risk allele odds ratio (ORs) = 2.09, 95% confidence interval (CI): 1.32-3.55, p = 0.0005). All six polymorphisms were found to have no significant effect on cellular radiosensitivity.

Conclusions

It is most likely that risk for radiation-induced fibrosis can be assessed by a combination of risk alleles. This finding needs to be replicated in further studies.     

Association of genetic polymorphisms of MK, IL-4, p16, p21, p53 genes and human gastric cancer in Taiwan

AIMS: To assess gastric cancer risk and clinical-pathological factors associated with genetic polymorphisms of MK, IL-4, p16, p21 and p53 genes. METHODS: A retrospective study was conducted for 123 patients who had recently developed primary gastric cancer. Clinical data and pathological findings were collected, genetic polymorphisms of MK, IL-4, p16, p21 and p53 genes were analysed, and the associations of genetic polymorphisms with gastric cancer carcinogenesis were evaluated. RESULTS: There was significant association of genetic polymorphisms between gastric cancer and control groups in p53 genes. After further stratification of the cancer group into different clinical-pathologic parameters, there were significant associations in the sex and LN involvement groups in MK gene; alcohol consumption group in p16 gene; age and cell differentiation groups in p21 gene; age and tumour location groups in p53 gene; but we fail to find any significant association with IL-4 gene polymorphisms. CONCLUSIONS: Genetic susceptibility testing is a tool to evaluate the association of genetic polymorphisms with gastric cancer carcinogenesis.