Objective: To establish a rat breast cancer cell line stably co-expressing mouse MIP-1α and B7-1,and to assay its in vitro biological activity.Methods: mMIP-1αcDNA was cloned into retrovirus vector pBabe puro to construct pBahe puro/mMIP-1α, then pBabe puro/mMIP-1α was used to transfect packaging cells and the anti-puromycin PA-317 packaging cells were proliferated. Meanwhile, pLXSN/mB7-1 was constructed and the anti-G418 cells were proliferated. Finally, the two supernatants were used to infect SHZ-88 together and the co-transfected cells were selected with 1 mg/ml G418 and 2 μg/ml puromycin together. Expression of mM1P-lα mRNA and protein in SHZ-88 and SHZ-88/mB7-1+mM1P-1α cells were analyzed by RT-PCR and immunocytochemistry, respectively. Expression of mB7-1mRNA and protein was analyzed by RT-PCR and flow cytometry, respectively. Lymphocyte proliferation activity of SHZ-88/B7-1+mM1P-1α was detected by MTT assay; chemotactic activity of MIP-1α was measured by chemotaxis assay. Results:A titer of 4.6×107 CFU/L was obtained after transfection with recombinant retroviral vector. The growth curve of cells showed that the recombinant retroviral had no effect on the growth of rat breast cancer cells. There was expression of B7-1 and MIP-1αmRNA/protein in SHZ-88/mMIP-1α+mB7-1 cells. The proliferation indices(PI) in mMIP-1α+mB7-1 group (1.95±0.31) was significantly higher than that in SHZ-88/PLXSN group (0.76±0.25)(P0.01). Chemotaxis assay showed that chemotactic activity of lymphocytes in mMIP-1α+mB7-1 group (3.88±0.33) was significantly higher than that in SHZ-88/pBabe puro group (0.99±0.19)(P0.001).Conclusion: A new rat breast cancer cel1 line SHZ-88/mMIP-1α+mB7-1 has been established, which can stably co-express MIP-1α and B7-1 gene and possesses biologic activity in vitro.

恶性肿瘤的发生常常是多个基因改变的结果,单一基因治疗的抗肿瘤效应往往十分有限,联合应用不同特性的细胞因子或趋化因子抗肿瘤是近年来的研究热点。本文着重综述趋化因子和B7分子在肿瘤基因治疗中的联合应用。B7分子是重要的共刺激分子,可为T细胞激活提供第二信号;趋化因子可通过趋化免疫活性细胞,部分趋化因子还可抑制肿瘤血管生成,从而产生抗肿瘤效应。共转染趋化因子和B7分子除通过“招引-活化”的机制外,还可抑制CD4+CD25+调节性T细胞亚型向肿瘤局部浸润,进而改善肿瘤局部免疫抑制状态;同时可抑制肿瘤血管生成,从而通过多个不同的途径,产生更强的抗肿瘤效应。但趋化因子具有抗肿瘤和促肿瘤双向性,因此选择合适的趋化因子与B7分子联合应用是至关重要的。

Gene transfer vector is the important ingredient in gene therapy,which includes virus vector and non-virus vector.At present,the mainly applied virus vectors are retroviral vector,adenovirus vector adenovirus-associated virus slow virus vector,herpes simplex virus vector and so on.Different viruses have their own advantages and disadvantages.With the increasing reform and consummation of virus vector, it can play its important role continuously in gene therapy by promoting its efficacy and safty in gene transfer.

患者女,52岁.患者2006年10月20日在外院行左侧甲状腺癌改良根治术,术后第2天起伤口引流出白色乳糜液,最大量达1 200mL/24h,于2006年11月13日转来我院.

Objective To evaluate the capability of the anti-tumor immunity induction by co- transfecting the breast cancer cells with MIP-1αand B7-1 gene.Methods SHZ-88 cells were inoculated in SD rats in order to establish rat breast cancer model.On day 10 after inoculation,the tumor size was documented,and rats were respectively vaccinated with SHZ-88,SHZ-88/B7-1,SHZ-88/MIP-1α,SHZ- 88/MIP-1α+B7-1 that were inactivated by mitomycin C on day 1 and 8.The therapeutic effect was tested in breast cancer SD rats by observing tumor volume and animal tumor-bearing life span in vivo.The im- mune functions in xenografts breast cancer rats were analyzed through checking the levels of whole blood CD4~+,CD8~+,CD4~+/CD8~+ ratio and the contents of serum IL-2 and IFN-γby flow cytometry and EIL- SA.Results The oncegenicity after subcutaneous inoculations of SHZ-88,B7-1,MIP-1αand MIP-1α+ B7-1 was 100%,30%,20% and 0 respectively,and the animal tumor-beating life span was prolonged in B7-1 group (63.80±1.64) d,MIP-1αgroup (64.20±1.92) d and MIP-1α+B7-1 group (89.00±2.55) d as compared with SHZ-88 group (53.40±1.14,P0.05).The peripheral blood CD4~+,CD8~+ and the ratio of CD4~+/CD8~+,and the contents of IL-2 and IFN-γin MIP-1α+B7-1 were the highest a- mong the groups.Fourteen,28 and 40 days after inoculation,the tumor size in MIP-1α+B7-1 group was less than other groups (P0.05).The antitumor effects had no significant difference after SHZ-88/MIP- 1α+B7-1 cells were injected into tumor center or contralateral axillary fossa (P0.05).Conclusion The co-transfection of MIP-1αgene and B7-1 gene could induce stronger anti-breast cancer effects.

<正>深静脉穿刺置管术是抢救重危病人、测定中心静脉压、长期静脉给药、胃肠道外营养支持治疗、化疗等常用的外科技术,在普通外科应用广泛。我科对胃肠道手术患者或危重患者常规行深静脉穿刺置管术,由于锁骨下静脉较颈内静脉易