白血病患者造血干细胞移植前单次全身照射的预后分析

Objective To analyze the prognostic factors of patients with leukemia treated with single fraction total body irradiation (SFTBI) followed by hernatopoietic stem cell transplantation (HSCT).Methods From January 2001 to September 2008, 102 patients received HSCT. The differences of the survival rate, relapse rate and incidence of interstitial pneumonia (IP) between groups regarding different genders, ages, pathological types, transplantation methods and TBI parameters were compared and the factors related with the survival rate, relapse rate and incidence of IP were analyzed. Results The followup time ranged from 15 to 1482 days (median, 406 days). The follow-up rate was 95.1%. 86 and 55patients were followed up more than one year and three years. The 1-and 3-year survival rates were 59.0%and 44.0%. In univariate analysis, the 3-year survival rate was signifcantly different between the groups with and without relapse before transplantation (20% vs. 55%, χ2 = 6.33, P = 0. 012), allogeneictranplantation versus autologous tranplantation (39% vs. 68%, χ2 = 8.06, P = 0.005), grade 3 or more acute graft versus host disease (aGVHD) and grade 0 -2 aGVHD (0% vs. 54%, χ2 = 7.52, P = 0.006),with and without relapse after transplantation (19% vs. 58%, χ2 = 10.13, P =0.001), with and without IP (23% vs. 58%, χ2 =8.35, P=0.004). Multivariate analysis showed that grade 3 or more aGVHD was the only statistically significant prognostic factors (χ2 = 12. 74 ,P =0. 000). The l-and 3-year relapse rateswere 30. 0% and 50. 0%. The incidence of relapse was obviously higher in the group with relapse before transplantation than that without (47% vs. 16%, χ2 =7. 32, P=0. 007). Multivariate analysis showed thatrelapse before transplantation was a significant factor predicting relapse after transplantation (χ2 = 9. 39,P =0. 020). The cumulative incidence of IP was 35.0%. The incidence of IP was different between groups with dose homogeneity > 3% and ≤ 3% (27% vs. 4%, χ2 = 5. 21, P = 0. 023), with and without acute parotitis (34% vs. 3%, χ2 = 14. 15, P= 0.000), allogeneic transplantation group and autologous transplantation group (31% vs. 8%, χ2= 7.70, P= 0.006). Multivariate analysis showed that transplantation methods, acute parotitis and dose homogeneity were statistically significant factors in predictingIP (χ2 = 10. 08 , 10. 08 and 7.69 , P = 0. 002 , 0. 002 and 0. 010 , respectively) . Conclusions Patients who develop grade 3 or higher aGVHD have poor prognosis. Dose homogeneity influences the incidence of IP. Patients undergoing allogeneic transplantation are apt to have IP. Acute parotitis is related with IP and might be a predictor.     

靶向VEGF发夹状RNA对宫颈癌HeLa细胞抑制作用的研究

目的:运用RNAi技术下调血管内皮生长因子(VEGF)在HeLa细胞中的表达,观察其对肿瘤细胞凋亡的影响,为人宫颈癌治疗提供理论依据。方法:设计并构建针对VEGF的携带绿色荧光蛋白(GFP)发夹状RNA(shRNA)质粒表达载体(PGPU6/GFP/Neo-shRNA),脂质体法转染HeLa细胞;荧光显微镜观察GFP的表达,并计算转染效率;RT-PCR检测HeLa细胞VEGF的表达,筛选出靶序列;再用流式细胞仪法检测细胞凋亡。结果:构建的PGPU6/GFP/Neo载体成功转入HeLa细胞;转染48h后,HeLa-shVEGF1组HeLa细胞VEGFmRNA表达的抑制率为75.0%;与HeLa组和HeLa-shNC组相比,HeLa-shVEGF1组HeLa细胞凋亡率明显增加,P<0.01。结论:本研究构建的PGPU6-shRNA表达载体携带GFP便于观察细胞的转染情况,且不影响U6启动子的转录,同时有效沉默了VEGF基因,明显增加HeLa细胞的凋亡,为未来肿瘤的治疗提供新途径。     

The MDR1 (P-glycoprotein) and MRP (P-190) transporters do not play a major role in the intrinsic multiple drug resistance of Jurkat T lymphocytes

The response of T cells in relation to the cell cycle has not been extensively studied. We have attempted to address this question using Jurkat T cells treated with cytostatic drugs known to arrest cells at various transition points of their cycle. We tested several concentrations of drugs that act at G1/S (hydroxyurea, lovastatin, thymidine), early S (aphidicolin, cyclosporin A, rapamycin) or G2+M (colchicine, nocodazole) in 24 h cultures. Cytofluorimetric analyses showed that cycling Jurkat cells were equally distributed between the G1 (44.9 ± 6.5%) and S (42.3 ± 8.0%) phases. Cell distribution in G2+M was 12.7 ± 2.8%. Hydroxyurea but not lovastatin increased the percentage of cells in S phase to ≈60–70% and both drugs decreased it to ≈30% in G1. Thymidine had no effects. Aphidicolin increased the distribution in S phase to ≈70% with a decrease in G1 to ≈30%. Cyclosporin A and rapamycin increased the percentage of the cells in G1 to ≈70% and decreased it to ≈25% in S phase. Nocodazole increased cell distribution in G2+M to ≈60% and induced a decrease in G1 to ≈10%. The effects of the drugs were not related to their toxicity and their limited efficiency raised the possibility that Jurkat cells possessed an intrinsic resistance to these xenobiotics. Time-course analysis showed (scanning electron microscopy) that the early morphological changes induced by colchicine were reversible. Drug efflux experiments (vinblastine) suggested that an ATP-dependent process could be involved. However, Northern blot analyses showed a weak signal for MDR1 (P-glycoprotein). In contrast, a probe for MRP (P-190) showed a strong signal in Jurkat and peripheral lymphocytes. The presence of drugs (cyclosporin A, nocodazole, thymidine) (24 h) did not upregulate its message and cell treatment with -butathione (S,R)-sulfoximine only moderately affected the efficiency of the glutathione S-conjugate MRP transporter. Our data suggest that the intrinsic multidrug resistance of leukemic Jurkat T cells does not appear to involve the MDR1 and MRP members of the ABC family of reverse drug transporters and these observations raise the possibility of the involvement of multifaceted mechanisms.     

158例非小细胞肺癌中3p25杂合性丢失研究

目的　探讨非小细胞肺癌组织 (NSCLC)中 3p2 5位点上等位基因的杂合性丢失 (LOH)与肺癌发生、发展的相关性。方法　采用银染聚合酶链反应结合二核苷酸 (CA)n重复程序出现多态性评价LOH ,分析 1 58例NSCLC癌组织中 3p2 5等位基因的LOH。结果　 1 58例NSCLC组织中 80例出现 3p2 5LOH ,总的杂合性丢失率为 50 .6%。腺癌组织中 3p2 5杂合性丢失率为 62 .0 % ,鳞癌为 43 .2 % ,两者间差异有显著性 (P <0 .0 5)。 3p2 5LOH与NSCLC临床分期关系不明显 ,Ⅰ、Ⅱ及Ⅲ期分别为 52 .2 %、45.0 %及 52 .8% ,4例良性肺肿瘤和 5例正常胎儿肺组织均未出现 3p2 5LOH。结论　本研究结果表明 3p2 5LOH在NSCLC中普遍存在 ,从而说明 3p2 5位点处可能存在着某些与肺癌发生、发展有关的抑癌基因     

Heterogeneity of Proliferative Activity in Nodule-in-Nodule Lesions of Small Hepatocellular Carcinoma

Surgically resected small hepatocellular carcinomas showing "nodule-in-nodule'formation were analyzed in terms of cell proliferative activity. The analysis was achieved by successful immunohistochemical demonstration of proliferating cell nuclear antigen in formalin-fixed paraffin-embedded tissue sections. Eight nodules (up to 3 cm in diameter) examined were either atypical adenomatous hyperplasia or hepatocellular carcinoma of low histologic grade, containing a discrete inner nodular area composed of obvious hepatocellular carcinoma of higher histologic grade. In all cases, the proliferating cell nuclear antigen labeling index of the latter area was much higher than that of the former, which in turn was slightly higher than that of the non-cancerous liver of the patient in 6 cases. The data presented here provide supporting evidence that the successive emergence and expansion of a more rapidly proliferating subclone within a nodule result in the stepwise progression of malignancy of human hepatocellular carcinoma.     

The PRB-dependent FOXO1/IGFBP-1 axis is essential for progestin to inhibit endometrial epithelial growth

Progestin inhibits the growth of normal and cancerous endometria via the progesterone receptor (PR), but the distinct functions and signalings of PR subtypes have not been fully understood. The aim of the present study was to dissect the key pathways of progestin to inhibit endometrial epithelial growth. Immortalized endometrial epithelial cells (EM-E6/E7/TERT) with stable PRA or PRB expression were established and used for the experiments. In vitro growth inhibition by progestin was mainly observed in EM-E6/E7/TERT cells with PRB rather than those with PRA. RT-PCR assay confirmed that FOXO1, a key gene for progestin action, was up-regulated by progestin in a PRB-dependent manner. cDNA microarray analysis identified IGFBP-1, which contains FOXO1 binding sites on its promoter, to be induced by medroxyprogesterone acetate (MPA) in EM-E6/E7/TERT cells with PRB but not with PRA. siRNA knockdown of FOXO1 disturbed the induction of IGFBP-1 by MPA, while IGFBP-1 knockdown showed no effect on MPA-induced FOXO1 expression, indicating that FOXO1 is an upstream regulator of IGFBP-1. Luciferase reporter assays showed that MPA activated the IGFBP-1 promoter, which was cancelled by FOXO1 knockdown. Chromatin immunoprecipitation assay confirmed the in vivo binding of FOXO1 to the core promoter of IGFBP-1. IGFBP-1 knockdown significantly attenuated the growth inhibitory effects of MPA. The FOXO1/IGFBP-1 axis is essential for PRB-dependent growth inhibition of endometrial epithelial cells, offering a potential therapeutic clue to enhance the progestin effect.     

造血干细胞移植治疗恶性血液病的临床观察

目的:观察我院骨髓移植治疗恶性血液病的疗效、探讨移植相关并发症的预防及处理等问题。方法:4例行HLA相合的同胞间异基因外周干细胞移植(Allo-PBSCT)、2例行自体外周干细胞移植(APBSCT)和1例行自体骨髓移植(Au-to-BMT)。结果:全部病例均成功造血重建。中性粒细胞≥0·5×109/L平均时间为11天;血小板≥20×109/L平均时间为16天;发生Ⅱ~Ⅲ度aGVHD1例,cGVHD1例,溶血性贫血2例,间质性肺炎2例,中位随访时间18(6~42)月,全部病例现均存活。结论:骨髓移植治疗恶性血液病有较好疗效。     

CD133 identifies perivascular niches in grade II–IV astrocytomas

The aim of the present study was to investigate the localization and distribution of the putative brain tumour stem cell marker CD133 in formalin fixed paraffin embedded astrocytomas. A retrospective analysis of 114 grade II, III and IV astrocytomas was undertaken. The immunohistochemical expression of CD133 in paraffin sections was analysed using morphometry. In all grades, CD133 was expressed on tumour and endothelial cells. Tumour cells were found in perivascular niches, as dispersed single cells and in pseudopalisade formations around necrosis. There was no correlation between the mean volume fraction of CD133⁺ niches and all CD133⁺ tumour cells and tumour grade. However, the volume fraction of CD133⁺ blood vessels increased significantly from 0.4% in diffuse astrocytomas to 2.2% in glioblastomas. Neither of them was related to patient survival. Double immunofluorescence stainings showed that the CD133⁺ niches both contained CD133⁺ cells with and without co-expression of the intermediate filament protein marker nestin, and only few CD133⁺/MIB-1⁺ proliferating cells were found. In conclusion, a CD133⁺ perivascular stem cell-like entity exists in astrocytomas. CD133⁺ tumour vessels may play an important role in a brain tumour stem cell context, while CD133 alone does not appear to be a specific tumour stem cell marker related to patient survival.