Expression of Hsp70-2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis

We investigated the possible role of Hsp70-2 in germ cell apoptosis induced by heat stress in monkey unilateral cryptorchid testis. The study focused on in situ analysis of the testicular cell DNA fragmentation and on the possible relationship between Hsp70-2 expression and germ cell apoptosis. The TUNEL result showed that most of the germ cells were labeled in the cryptorchid testis on d 5 after induction of cryptorchidism; that with most of the apoptotic germ cells depleted, only a few germ cells were labeled on d 10; and that almost no apoptotic signal was observed in the cryptorchid testis on d 15 and thereafter. This indicates that the increasing germ cell degeneration in cryptorchid testis may take the form of apoptosis. Using in situ hybridization, immunohistochemistry, and Northern blot, we examined the changes of Hsp70-2 expression in the monkey cryptorchid testis. The level of Hsp70-2 mRNA decreased slightly, while the expression of HSP70-2 protein was almost unchanged at the early stage of germ cell apoptosis in the cryptorchid testis on d 5 and dropped dramatically along with the loss of apoptotic germ cells in the cryptorchid testis on d 10 after operation. It is therefore suggested that Hsp70-2 might not take part in inhibiting the apoptosis of germ cells at the early stage during operation-induced cryptorchid testis, and that Hsp70-2 gene does not belong to the immediate early related gene responsible for germ cell apoptosis induced by heat stress.     

Anti-cardiolipin and Anti-β2-glycoprotein I Antibodies in Patients with Inflammatory Bowel Disease

Patients with inflammatory bowel disease (IBD)frequently suffer from thromboembolic events.Anti-cardiolipin (aCL) antibodies have been shown to beassociated with thrombosis. Recently, the antibodies against the anti-cardiolipin cofactor₂-glycoprotein I(a₂GPI) have been found with higherspecificity for thrombosis. The presence of theseantibodies was assessed in 128 patients with IBD [83 with ulcerative colitis (UC) and 45 withCrohn's disease (CD)] and 100 healthy controls (blooddonors). Patients with UC and CD had a significantlyhigher prevalence of aCL (18.1% and 15.6%, respectively) than healthy controls (HC) (3%). Eleven IBDpatients (8.6%) but no HC had a₂GPI.None of the IBD patients with a history of thrombosishad aCL and only one of them (a UC patient with deepvein thrombosis of the right leg) had a high titer of IgGa₂GPI. In conclusion, these data showthat both aCL and a₂GPI aresignificantly associated with IBD but further studiesare needed to determine the significance of our findings.     

HSP70-肿瘤肽复合物的纯化及其对肝癌细胞株HepG-2增殖的作用

目的:应用快速蛋白液相色谱(FPLC)系统从肝癌组织中分离和纯4EHSP70-肽复合物,并研究其对肝癌细胞系HepG-2增殖的影响．方法:将组织进行匀浆、高速离心提取总蛋白后依次进行ConA-Sepharose亲和层析和 DEAE-Sephacel子交换层析分离纯化,所得蛋白经SDS-聚丙烯酰胺凝胶电泳和Western blot进行蛋白分子质量及性质鉴定,Bradford 法测定蛋白浓度;利用MTT方法检测HSP70- 肽复合物对HepG-2细胞增长的情况．结果:分离、纯化得到的蛋白经SDS-聚丙烯酰胺凝胶电泳、考马斯量蓝鉴定为单一带, 分子质量为70 kDa;Western blot结果证实为 HSP70,每10 g组织最终获得1．5 mg的HSP70; HSP70刺激组12,48,72 h与对照组的A值之间有显著差异(0．1 mg／L:t=-0．2500,P=0．00; t=-0．1777,P=0．001:t=-0．3094,P=0．001; 0．5 mg／L:t=-0．2878,P=0．00;t=-0．2044,P= 0．00;t=-0．3285,P=0．00;1 mg／L:t=-0．3118, P=0．00;t=-0．2592,P=0．00;t=-0．1994, P=0．025;5 mg／L:t=-0．4007,P=0．00;t= -0．1302,P=0．016;t=-0．2537,P=0．005),细胞存活率明显高于对照组(P<0．01)．结论:使用本分离纯化方法可获得高纯度 HSP70肽复合物;HSP70肽复合物可以促进 HepG-2细胞的生长．     

Effect of a gonadotrophin-releasing hormone analogue on lung function in lymphangioleiomyomatosis

BACKGROUND: Lymphangioleiomyomatosis (LAM), a multisystem disease occurring primarily in women, is characterized by cystic lung destruction, and kidney and lymphatic tumors, caused by the proliferation of abnormal-appearing cells (ie, LAM cells) with a smooth muscle cell phenotype that express melanoma antigens and are capable of metastasizing. Estrogen receptors are present in LAM cells, and this finding, along with reports of disease progression during pregnancy or following exogenous estrogen administration, suggest the involvement of estrogens in the pathogenesis of LAM. Consequently, antiestrogen therapies have been employed in treatment. The goal of this prospective study was to evaluate the efficacy of triptorelin, a gonadotrophin-releasing hormone analogue, in 11 premenopausal women with LAM. METHODS: Patients were evaluated at baseline and every 3 to 6 months thereafter, for a total of 36 months. Hormonal assays, pulmonary function tests, 6-min walk tests, high-resolution CT scans of the chest, and bone mineral density studies were performed. RESULTS: Gonadal suppression was achieved in all patients. Overall, a significant decline in lung function was observed; two patients underwent lung transplantation 1 year after study enrollment, and another patient was lost to follow-up. Treatment with triptorelin was associated with a decline in bone mineral density. CONCLUSIONS: Triptorelin appears not to prevent a decline in lung function in patients with LAM. Its use, however, may be associated with the loss of bone mineral density.     

CD27 costimulation contributes substantially to the expansion of functional memory CD8+ T cells after peptide immunization

Naive T cells require signals from multiple costimulatory receptors to acquire full effector function and differentiate to long‐lived memory cells. The costimulatory receptor, CD27, is essential for optimal T‐cell priming and memory differentiation in a variety of settings, although whether CD27 is similarly required during memory CD8⁺ T‐cell reactivation remains controversial. We have used OVA and anti‐CD40 to establish a memory CD8⁺ T‐cell population and report here that their secondary expansion, driven by peptide and anti‐CD40, polyI:C, or LPS, requires CD27. Furthermore, antigenic peptide and a soluble form of the CD27 ligand, CD70 (soluble recombinant CD70 (sCD70)), is sufficient for secondary memory CD8⁺ T‐cell accumulation at multiple anatomical sites, dependent on CD80/86. Prior to boost, resting effector‐ and central‐memory CD8⁺ T cells both expressed CD27 with greater expression on central memory cells. Nonetheless, both populations upregulated CD27 after TCR engagement and accumulated in proportion after boosting with Ag and sCD70. Mechanistically, sCD70 increased the frequency of divided and cytolytic memory T cells, conferred resistance to apoptosis and enabled retardation of tumor growth in vivo. These data demonstrate the central role played by CD27/70 during secondary CD8⁺ T‐cell activation to a peptide Ag, and identify sCD70 as an immunotherapeutic adjuvant for antitumor immunity.     

A 14-mer Peptide from HSP70 Protein is the Critical Epitope Which Enhances NK Activity Against Tumor Cells in vivo

Heat shock protein 70 (Hsp70) has been found to play key roles in tumor immunity due its chaperone function of binding antigenic peptides. Here we report it can also stimulate NK cells in vivo, which is another role in Hsp70s' anti-tumor response. Injecting Hsp70 into mice increased splenic NK cell populations, which may be reason for anti-tumor effect of Hsp70. The Hsp70 14-mer peptide (aa450–463, TRD) was identified as the critical epitope for this stimulatory activity. It was the murine Hsp70 14-mer peptide TRD instead of the corresponding human Hsp70 14-mer peptide TKD that functioned in the mouse experimental model.     

Expression of heat shock proteins 70 and 47 in tissues following short-pulse laser irradiation: Assessment of thermal damage and healing

In order to develop effective laser-based therapeutics, the extent of laser-induced damage must be quantified for given laser parameters. Therefore, we want to determine the spatiotemporal expression patterns of heat shock proteins, both to understand the roles of heat shock proteins in laser-induced tissue damage and repair and to develop heat shock proteins as tools to illustrate the extent of laser-induced damage and wound healing following irradiation. We exposed anesthetized mice to the focused beam of a short-pulse Nd:YAG laser (1064 nm; 200 ns pulsewidth) for 15 s, while measuring temperature distribution in the skin using an infrared thermal camera. Following irradiation, we examined expression of HSP47 and HSP70 over time (0–24 h) as indicators of the heat shock response and recovery from damage in the laser-irradiated region. Expression patterns of HSP70 and HSP47 as detected by immunohistochemistry and confocal microscopy delineate the extent of damage and the process of healing in tissue. Both HSP70 and HSP47 were expressed in dermis and epidermis following laser irradiation, and the spatial and temporal changes in HSP expression patterns define the laser-induced thermal damage zone and the process of healing in tissues. HSP70 may define biochemically the thermal damage zone in which cells are targeted for destruction, and HSP47 may illustrate the process of recovery from thermally induced damage. Studying the effects of different laser parameters on the expression of HSPs will allow development of effective laser therapies that provide accurate and precise tissue ablation and may promote rapid wound healing following laser-based surgery.     

新中国70周年医院药学的发展历程与趋势(Ⅱ)

3第三阶段:药学服务,创新驱动国内外药学界普遍认为,未来医院药学应以患者健康为目标、“药学服务”为重点的药学专业服务工作。20世纪90年代中期,药学服务的理念传入我国,上海长海医院药学部胡晋红教授提出的“全程化药学服务”(integrated pharmaceutical care)概念获得普遍认同。在这个框架内,药师与患者、医护人员及其他专业人员进行合作,药师增加对药物使用监控的职能,以及为达到改善患者健康和生活质量这个特定目标而提供服务。药学服务包括用药前的宣传教育,用药过程中的顾问、监测及用药后的监测与评价[5]。20世纪90年代至今,医院药学处于药学服务阶段,以现代药学及临床药学为支柱,支持这一工作的基础学科为药物治疗学、临床药学及计算机网络化管理技术[1]。