表皮生长因子和甲硫氨酸脑啡肽诱导人白细胞c—fos基因表达的研究

由健康人外周血分离淋巴细胞及溶血处理后的全白细胞，以外源性表皮生长因子和甲硫氨酸脑啡肽培育后取细胞液帛晟滴片和滴膜，用生物素标记的ｃ－ｆｏｓｃＤＮＡ探针进行原位杂交和斑点印迹杂交。结果显示表皮生升因子和甲硫氨酸脑啡肽两组ｃ－ｆｏｓ原癌基因的表达对照组增。     

Nitric Oxide Synthase Expression in Macrophages of Histoplasma capsulatum-Infected Mice Is Associated with Splenocyte Apoptosis and Unresponsiveness

Splenic macrophages from Histoplasma capsulatum-infected mice express inducible nitric oxide synthase (iNOS), and the iNOS expression correlates with severity of the infection. We examined whether production of NO is responsible for apoptosis and the anti-lymphoproliferative response of splenocytes from mice infected with H. capsulatum. In situ terminal deoxynucleotidyl transferase nick end labeling revealed apoptotic nuclei in cryosections of spleen from infected but not normal mice. Splenocytes of infected mice were unresponsive to stimulation by either concanavalin A or heat-killed H. capsulatum yeast cells. Splenocyte responsiveness was restored by addition to the medium of N^G-monomethyl-l-arginine, a known inhibitor of NO production. The proliferative response of splenocytes from infected mice was also restored by depletion of macrophages or by replacement with macrophages from normal mice. In addition, expression of iNOS returned to its basal level when the animals had recovered from infection. These results suggest that suppressor cell activity of macrophages is associated with production of NO, which also appears to be an effector molecule for apoptosis of cultured splenocytes from infected mice.

Nitric oxide (NO) has been reported to induce apoptosis in many cells including smooth muscle cells (20), oligodendrocytes (27), pancreatic β cells (11), melanoma cells (35), thymocytes (7), B lymphocytes (4), and macrophages (2). Fehsel et al. recently demonstrated apoptosis in freshly isolated thymocytes after exposure to NO (7). In the same report, they also showed apoptotic foci in close proximity to blood vessels after lipopolysaccharide treatment. Capillary endothelial and dendritic cells adjacent to apoptotic foci stained strongly for inducible nitric oxide synthase (iNOS), suggesting that NO may be the mediator for thymic apoptosis (7). Data from another laboratory also showed that cloned thymic stromal cell monolayers eliminate thymocytes in vitro through production of NO (26). Furthermore, apoptosis has been suggested as a mechanism by which the immune system replenishes itself and maintains homeostasis (30).The dimorphic fungus Histoplasma capsulatum is a facultative intracellular pathogen of the macrophage (32). Although it is not an obligate intracellular pathogen, the organism is found almost exclusively inside host cells during histoplasmosis (5). In our in vitro studies, H. capsulatum exhibits uninhibited growth in normal unstimulated murine macrophages (32). In activated macrophages, either peritoneal macrophages and cells from the Raw 264.7 line stimulated by gamma interferon (IFN-γ) or splenic macrophages stimulated by IFN-γ and lipopolysaccharide, growth of the fungus is inhibited (13, 18, 32). Furthermore, the anti-histoplasma activity of macrophages is dependent on the expression of iNOS and the production of NO (14, 18). However, the significance of NO production in immunoregulation of histoplasmosis is not clearly defined.In this study, we examined whether NO can act as a regulator of apoptosis in lymphoproliferative responses of splenocytes from H. capsulatum-infected mice. We showed that iNOS was induced in splenic macrophages during active infection and the expression of iNOS coincided with active infection. We also observed by in situ terminal deoxynucleotidyl transferase (TdT) nick end labeling (TUNEL) of spleen sections that apoptosis occurred in immune cells in the spleens of infected mice but was minimal in control mice. The link between apoptosis and NO production was established by inclusion of N^G-monomethyl-l-arginine (NMMA) in the culture medium. Inhibition of NO production reduced the amount of apoptosis in splenocyte culture. Thereby, we also confirmed the findings of Zhou et al. (36) that production of NO by splenocytes of H. capsulatum-infected mice suppressed the splenic lymphocyte proliferative response. In addition, we showed that macrophages were mediators of splenocyte unresponsiveness through the NO that they produced and that NO production was associated with apoptotic changes in cultured splenocytes from infected mice.     

甲型副伤寒沙门氏菌体抗原抗血清的简易提纯

目的 单克隆抗体以其结合抗原的专一性用于血清学检验是一种必然趋势,而多克隆机体则以其简易的制备方式在目前仍得到广泛的应用,但多克隆抗体的提纯仍有许多繁琐之处,使其优势受到影响。本试验以甲型副伤寒沙门氏菌体抗原多克隆抗体的制备及提纯为例,提出可行的更简易的提纯方法。方法 利用产G蛋白链球菌,以一种较为简易的特异性结合和解离的方法来纯化甲型副伤寒沙门氏菌体抗原免疫血清。结果 得到了特异性IgG抗体。结论 利用这种纯化方法在实验室简易可行。     

癌瘤患者骨髓细胞c－erbB和c－erbA的基因异常

本文在大系列研究造血系统恶性肿瘤-白血病和白血病前期的基础上，对３１例恶性肿瘤患者骨髓进行血液学、细胞和分子遗传学研究。发现恶性肿瘤患者骨髓有与白血病前期骨髓相似的病态造血（Ｍｙｅｌｏｄｙｓｐｌａｓｉａ）和相关基因异常即ｃ－ｅｒｂＢ基因重排、扩增和ｃ－ｅｒｂＡ缺失。它们不仅同骨髓红系病态造血相关，也同巨核系病态造血相关。骨髓巨核系病态造血及其相关基因异常可能代表急性淋巴细胞白血病等多种恶性肿瘤的早期病变。而ｃ－ｅｒｂＢ重排及其相关的骨髓巨核系异常增生可能是肿瘤发病的遗传背景。     

分化抗原5C5在人免疫细胞的表达

目的　明确 5C5蛋白为分化抗原 ,检测其在人免疫细胞的表达 ,以及它的信号传导作用。方法　用 5C5蛋白免疫小鼠 ,制备抗 5C5蛋白的单抗。用反义技术制备 5C5蛋白表达抑制的 3D5细胞。用免疫荧光染色和Western印迹确定 5C5单抗的特异性。用胞膜蛋白免疫沉淀和流式细胞术确定其分化抗原的性质。用双色免疫荧光染色流式细胞术检测 5C5分化抗原在多种人免疫细胞的表达。用Fura 3 AM试剂作探针 ,用激光共聚焦显微镜观察胞内Ca2 + 浓度的变化。结果　制备了抗 5C5蛋白的特异性单抗。用 5C5单抗作探针 ,从 3D5细胞膜免疫沉淀到 1条相对分子质量 (Mr)为 19× 10 3的蛋白带 ,并在活 3D5细胞见阳性免疫荧光染色。 5C5分化抗原在人周围血CD19+ 、CD14 + 、CD4 + 、CD8+ 和CD5 6 + 细胞的表达率分别为 (33.4 7± 7.9) %、(86 .13± 6 .7) %、(15 .79± 7.1) %、(18.11±12 .77) %和 (11.76± 7.4 7) % ,在人免疫细胞株的表达率分别为 5 8.0 % (Nalm6 )、72 .7% (3D5 )、5 3.6 %(BJAB)、6 1.0 % (Daudi)、6 .9% (SKW6 )、6 .3% (Jurkat)、5 .8% (Peer)、2 1.3% (K5 6 2 )和 39.1% (U937)。3D5细胞在 5C5单抗刺激下 ,胞内Ca2 + 浓度逐渐增高 ,可达 2倍左右 ,对照小鼠Ig则无此作用。结论5C5蛋白为一个分化抗原 ,在人周     

Synthesis and characterization of amphiphilic and microphase-separated graft copolymers, 1. Synthesis and bulk characterization of polystyrene-graft-ω-stearyl-poly(ethylene oxide)

Using the rigid and hydrophobic polystyrene (PS) chain as backbone, onto which flexible and hydrophilic stearyl-poly(ethylene oxide) (SPEO) chains are grafted, a new kind of amphiphilic, microphase-separated graft copolymer was synthesized using the macromonomer technique. Stearyl-poly(ethylene oxide) macromonomers with acryloyl end-group (SPEO-A) were prepared through an end-group exchange reaction of α-stearyl-ω-hydroxypoly(ethylene oxide) (SPEO-OH) and acryloyl chloride in the presence of triethylamine. The radical copolymerization of styrene with SPEO-A was carried out under various experimental conditions. Following a careful examination of their purity, the structure of the prepared copolymers was characterized by means of IR, ¹H NMR and GPC analyses. A new feasible method using first derivative UV spectrometry was developed for quantitative determination of the bulk composition of the graft copolymers. Copolymers with a wide range of bulk composition and satisfactory grafting degree were obtained.     

磷脂双层膜稳定性的理论分析

目的：越来越多磷脂双层膜的原子力显微镜扫描力谱显示溶液离子浓度对磷脂双层膜稳定性有很大影响，本文从理论角度研究溶液离子浓度对磷脂双层膜稳定性的影响，同时解释两个基于不同模型模拟结果差别的原因。方法：主要根据磷脂分子极性端带电特性，将极性端看作偶极子，然后基于偶极-偶极相互作用理论以及泊松-玻尔兹曼理论解决以上问题。结论：随着离子浓度的增加，磷脂双层膜趋于更加稳定；基于两个不同模型模拟结果差别的原因是高浓度溶液的强静电屏蔽作用。     

食管分层应力-应变研究

目的　探讨食管壁各层环向应力 应变分布特征。方法　将 8只Wister大鼠的食管标记、测量在体长度后剪下置于去钙离子Krebs液中。剪下标记的中段食管用做充压试验 ,将其在显微镜下分离成内层食管 (粘膜和粘膜下层 )和外层食管 (固有肌层 )。充压时全层和分层食管均被拉长至在体长度 ,以每分钟升高 2cmH2 O速度匀速充压。根据所测图像的几何数据计算全层和分层食管的Kirchhoff应力和Green应变。零应力状态作为计算应力和应变的基本参照状态。结果　 (1 )食管离体后较在体长度缩短 2 4%。 (2 )全层食管和分层食管的环向应力 应变曲线均符合指数方程τ =(τ +β)eα(ε-ε ) -β(判定系数 >0 .96) ,为非线性关系。 (3 )与全层食管相比 ,分层后内层食管应力 应变曲线较陡直 ,且移向左侧 ,差别有统计意义 (P <0 .0 5 ) ,外层食管的曲线移向右侧 ,差别无统计意义 (P >0 .0 5 ) ,内外两层食管的应力 -应变曲线亦有统计差别 (P <0 .0 5 )。结论　食管壁各层环向应力 -应变均符合非线性关系 ,食管内层不如外层易变形     

Association of genetic polymorphisms and haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma

OBJECTIVE: To explore the relationship of the genetic polymorphisms and the haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma (PTC) in Chinese Hans. METHODS: A hospital based 1:1 matched case-control study was carried out. The polymorphisms for 204 pairs of PTC cases and healthy controls were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific oligonucleotide (PCR-ASO) assays. RESULTS: (1) The PTC risk was marginally increased in the hMLH1 1151TA genotype, with odds ratio (OR) of 2.15 (95%CI: 0.99-4.85); the PTC risk was significantly increased in the mutant genotype 1151TA+AA, with OR of 2.15 (95%CI: 1.02-4.69); (2) The haplotypes of -93G, 1151A, 655A in the hMLH1 gene could increase the PTC risk, with OR of 2.67 (95%CI: 1.16-6.53, P=0.011), compared with the haplotype of -93G, 1151T, 655A; (3) Compared to 3124A, 2835G haplotype in hMSH3 gene, the 3124G, 2835A haplotype could increase the PTC risk marginally, with OR of 3.08 (95%CI: 0.92-13.25). CONCLUSION: The 1151T/A polymorphism in hMLH1 was associated with PTC; both the haplotype of -93G, 1151A, 655A in hMLH1 and the 3124G, 2835A haplotype in hMSH3 were associated with PTC.