靶向性自杀基因治疗系统联合放射线治疗前列腺癌移植瘤的实验研究

目的初步判断针对整合素αv的靶向性自杀基因治疗系统(RGD4C AAVP HSV-TK/ GCV)是否能提高前列腺癌细胞株DU145移植瘤的放射效应。方法建立移植性前列腺癌细胞株DU145的裸鼠肿瘤模型48只。当位于每只裸鼠右大腿的肿瘤直径达6.0mm(5.8～6.3 mm)时,开始实验。实验共分成6组(每组8只),分别为对照组、单纯放射组、单纯RGD-4C AAVP HSV-TK/GCV组(RGD-4C组)、单纯AAVP HSV-TK/GCV组(无RGD-4C组)、RGD-4C AAVP HSV-TK/GCV联合放射组(XRT RGD4C组)和AAVP HSV-TK/GCV联合放射组(XRT 无RGD-4C组)。观察指标为肿瘤生长延迟时问(肿瘤从6.0 mm生长至12.0 mm所需时间)和肿瘤治愈情况。结果除5只在实验过程中死亡外,单纯放射组、RGD-4C组、无RGD-4C组各有1只和XRT RGD-4C组3只肿瘤被治愈。统计分析余下37只肿瘤生长情况发现,RGD-4C组、无RGD-4C组和单纯放射组的绝对延迟时间分别为(24.4±9.0)、(22.6±11.3)和(28.3±5.5)d;当RGD4C AAVP HSV-TK/GCV和AAVP HSV-TK/ GCV分别联合放射时,其绝对延迟时间分别为(64.7±23.8)和(35.4±9.6)d,而标准化的延迟时间则分别为(40.3±23.8)和(12.8±9.6)d,它们对放射的增益因子分别为1.42和0.45。结论针对整合素αv的靶向性自杀基因治疗系统RGD-4C AAVP HSV-TK/GCV能显著提高前列腺癌细胞株DU145移植瘤的放射效应,值得进一步研究。     

差异表达基因整合素a2白细胞介素-8Cx43在早期宫颈鳞癌中的表达及意义

目的 研究早期宫颈鳞癌差异表达基因整合素d2、白细胞介素-8（IL-8）和间隙连接蛋白43（Cx43）在宫颈鳞癌中的表达及其与淋巴结转移的关系，探讨其在宫颈鳞癌淋巴结转移中的作用机制。方法 采用反转录-聚合酶链反应（RT—PCR）、组织芯片进行的免疫组织化学方法检测早期宫颈鳞癌组织中整合素d2、IL-8和Cx43mRNA及蛋白的表达。结果有淋巴结转移的早期宫颈鳞癌组织中整合素d2和IL-8的表达显著高于非转移组织（P〈0．05），CX43的表达则相反（P〈0．01）。且整合素d2和IL-8的表达呈正相关（r=0．241，P〈0．05）。结论整合素d2、IL-8和CX43与早期宫颈鳞癌的淋巴结转移密切相关，在一定程度上发挥了促进和抑制早期宫颈鳞癌淋巴结转移的作用。     

Inhibition of melanoma cell motility by the snake venom disintegrin eristostatin.

Eristostatin, an RGD-containing disintegrin isolated from the venom of Eristicophis macmahoni, inhibits lung or liver colonization of melanoma cells in a mouse model. In this study, transwell migration and in vitro wound closure assays were used to determine the effect of eristostatin on the migration of melanoma cells. Eristostatin significantly impaired the migration of five human melanoma cell lines. Furthermore, it specifically inhibited cell migration on fibronectin in a concentration-dependent manner, but not that on collagen IV or laminin. In contrast, eristostatin was found to have no effect on cell proliferation or angiogenesis. These results indicate that the interaction between eristostatin and melanoma cells may involve fibronectin-binding integrins that mediate cell migration. Mutations to alanine of seven residues within the RGD loop of eristostatin and four residues outside the RGD loop of eristostatin resulted in significantly less potency in both platelet aggregation and wound closure assays. For six of the mutations, however, decreased activity was found only in the latter assay. We conclude that a different mechanism and/or integrin is involved in these two cell activities.     

肠道淋巴细胞归巢受体在胃肠功能障碍的危重症患儿外周血中表达的变化

目的 探讨危重症患儿胃肠功能障碍时外周血肠道淋巴细胞归巢受体(整合素α4β7和L-选择素)的表达及其与肠道黏膜免疫功能的关系.方法 4个月～14岁危重症患儿45例,分为胃肠功能衰竭组及非胃肠功能衰竭组,选择25例健康体检儿为对照组.用流式细胞仪检测整合素α4β7和L-选择素的表达.结果 胃肠功能衰竭组外周血中整合素α4β7和L-选择素水平明显减低,胃肠功能衰竭组与非胃肠功能衰竭组比较差异有显著性(P<0.05),胃肠功能衰竭组与对照组比较差异亦有显著性(P<0.05).结论 整合素α4β7和L-选择素水平在危重症患儿发生胃肠功能障碍的早期就降低.肠道淋巴细胞归巢受体可能通过抑制淋巴细胞向肠道的归巢,使肠黏膜免疫屏障受损,进而降低肠黏膜的免疫功能.     

胚胎着床及其分子调控研究进展

胚胎着床是哺乳动物特有的生殖生理现象,涉及到胚胎和母体卵巢、子宫之间复杂的相互作用和分子信号联系。各种动物在胚胎着床的方式及分子调控机制上各有不同,目前已发现包括由胚胎和子宫内膜分泌合成的多种生殖激素、生长因子、细胞黏附分子和细胞外基质等物质参与了胚胎着床的分子调控过程。本文综述近年来在人类及相关动物胚胎着床的研究进展,并重点阐述在胚胎着床中存在于胚胎滋养层表面与母体子宫内膜上皮表面作用的相关信号联系及分子调控机制,以期对提高妊娠率、防止胎儿早期流产以及开发简便高效的临床避孕措施的研究提供理论依据和借鉴。     

整合素β3和细胞间隙连接蛋白43与体外受精-胚胎移植妊娠结局的关系

目的:探讨子宫内膜整合素β3、细胞间隙连接蛋白43表达水平与体外受精-胚胎移植妊娠结局的关系,进而分析其与子宫内膜容受性的相关性。方法:前瞻性收集了IVF/ICSI-ET病例36例,其中11例临床妊娠者作为妊娠组,25例未妊娠者作为非妊娠组。于黄体中期取内膜,应用单克隆抗体免疫组化方法检测子宫内膜整合素β3和Cx43表达情况。结果:妊娠组与非妊娠组子宫内膜腔上皮整合素β3表达的HSCORE平均分数分别为0.55(0.00~1.60)和0.07(0.00~1.10),差异有显著性(P=0.02);而子宫内膜腺上皮整合素β3表达比较,两组之间差异无显著性;子宫内膜间质细胞和上皮细胞中Cx43表达在两组均未发现统计学差异。结论:子宫内膜腔上皮整合素β3表达水平可能与体外受精-胚胎移植妊娠结局有相关性。     

Robust Micromechanical Neurite Elicitation in Synapse-Competent Neurons Via Magnetic Bead Force Application

The ability to engineer living networks of interconnected neurons with specified connectivity would facilitate the study of synaptogenesis and information processing in the nervous system. Previously, we found that a neurite can be elicited from embryonic chick forebrain neurons by direct mechanical means using magnetic bead force application (MBFA); however, our previous studies and others focused on young, synapse-incompetent neurons. To address this issue, we tested cultures of embryonic chick forebrain neurons of varying age and found that neurites could be micromechanically elicited via MBFA at all ages tested, which ranged between 7 and 22 embryonic equivalent (EE) days (days in ovo plus days in vitro). The probability of neurite initiation was at least 40% for all ages, with a maximum of ∼80% after 2–4 days in vitro, and a decrease to ∼60% by day 10 in vitro. The force required to elicit a neurite was ∼1500 pN with a minimum of ∼700 pN at embryonic equivalent day 14. The probability of success was similar for two rates of force application (10 and 500 pN/s). Neurite initiation via micromechanical force is robust with respect to cell age, and micromechanical force can induce neurites in synapse-competent neurons.     

Comparison of effects of methylprednisolone and anti-CD11d antibody treatments on autonomic dysreflexia after spinal cord injury

Autonomic dysreflexia is a condition of episodic hypertension that develops after spinal cord injury (SCI). We previously showed that a two-day anti-inflammatory treatment with an anti-CD11d integrin monoclonal antibody (mAb), soon after SCI in rats, reduced the magnitude of dysreflexia for at least 6 weeks. Effects of methylprednisolone (MP), a commonly used neuroprotective treatment for SCI, on dysreflexia have never been examined. We compared the effects of a 2-day MP treatment and/or the anti-CD11d mAb on autonomic dysreflexia, elicited by colon distension, after clip-compression SCI at the 4th thoracic segment (T4) in rats. We assessed the effects of each treatment on the size of the calcitonin gene-related peptide (CGRP)-immunoreactive afferent arbour in the dorsal horn, as changes in this arbour can correlate with the development of dysreflexia. MP reduced autonomic dysreflexia by approximately 50% at 2 weeks after SCI, but this effect was lost by 6 weeks. At 2 weeks, the combined effects of MP and the mAb were not additive, reducing dysreflexia by approximately 50%. Neither MP nor the mAb treatment altered the area of CGRP-immunoreactive fibres in the lumbar cord, the crucial input region for dysreflexia initiated by colon distension. However, both treatments led to increased fibre areas in the T9 segment, correlated with greater tissue integrity and smaller lesions, delineated by inflammatory cells. In summary, MP only temporarily decreases autonomic dysreflexia after SCI. The early beneficial effects of both treatments on dysreflexia do not relate to changes in the CGRP-immunoreactive afferent arbour but may correlate with decreased lesion progression.     

Fibrinogen-CD11b/CD18 interaction activates the NF-kappa B pathway and delays apoptosis in human neutrophils

The regulation of neutrophil half-life by members of the coagulation cascade is critical for the resolution of the inflammatory response. We have demonstrated that soluble fibrinogen (sFbg) delays human neutrophil (PMN) apoptosis through a mechanism that involves CD11b interactions, and phosphorylation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase 1/2 (ERK1/2).Since NF-kappa B is a key element in the regulation of apoptotic mechanisms in several immune cells, we investigated whether NF-kappa B is involved in the control of PMN survival by sFbg. We show that sFbg triggers inhibitor protein kappa B (I kappa B-alpha) degradation and NF-kappa B activation. Furthermore, pharmacological inhibition of NF-kappa B abrogates sFbg effects on apoptosis. In addition, specific inhibition of MAPK ERK1/2 significantly reduces NF-kappa B translocation by sFbg, suggesting a relationship between ERK1/2 and NF-kappa B activation. Similar results are obtained when granulocytic-differentiated HL-60 cells are treated with sFbg, making this model highly attractive for integrin-induced gene expression studies. It can be concluded that NF-kappa B participates in the prevention of apoptosis induced by sFbg with the participation of MAPK ERK1/2. These results shed light on the molecular mechanisms that control human granulocyte apoptosis, and suggest that NF-kappa B regulation may be of benefit for the resolution of the inflammatory response.