Resistance to morphine analgesic tolerance in rats with deleted transient receptor potential vanilloid type 1-expressing sensory neurons

Deletion of transient receptor potential vanilloid type 1 (TRPV1)-expressing afferent neurons reduces presynaptic mu opioid receptors but paradoxically potentiates the analgesic efficacy of mu opioid agonists. In this study, we determined if removal of TRPV1-expressing afferent neurons by resiniferatoxin (RTX), an ultrapotent capsaicin analog, influences the development of opioid analgesic tolerance. Morphine tolerance was induced by daily intrathecal injections of 10 microg of morphine for 14 consecutive days or by daily i.p. injections of 10 mg/kg of morphine for 10 days. In vehicle-treated rats, the effect of intrathecal or systemic morphine on the mechanical withdrawal threshold was gradually diminished within 7 days. However, the analgesic effect of intrathecal and systemic morphine was sustained in RTX-treated rats at the time the morphine effect was lost in the vehicle group. Furthermore, the mu opioid receptor-G protein coupling in the spinal cord was significantly decreased ( approximately 22%) in vehicle-treated morphine tolerant rats, but was not significantly altered in RTX-treated rats receiving the same treatment with morphine. Additionally, there was a large reduction in protein kinase Cgamma-immunoreactive afferent terminals in the spinal dorsal horn of RTX-treated rats. These findings suggest that loss of TRPV1-expressing sensory neurons attenuates the development of morphine analgesic tolerance possibly by reducing mu opioid receptor desensitization through protein kinase Cgamma in the spinal cord. These data also suggest that the function of presynaptic mu opioid receptors on TRPV1-expressing sensory neurons is particularly sensitive to down-regulation by mu opioid agonists during opioid tolerance development.     

帕罗西汀合并认知疗法治疗酒依赖伴发抑郁及对戒酒的影响

目的评价帕罗西汀合并认知疗法治疗酒依赖伴发抑郁的疗效及对戒酒的影响。方法将98例酒依赖伴发抑郁的患者随机分为研究组和对照组,分别用认知疗法合并帕罗西汀、单用帕罗西汀治疗4个月。用汉密顿抑郁量表(HAMD)评定疗效;用复饮率评定戒酒效果。结果治疗1个月后,研究组HAMD评分16.79±6.50与对照组相近17.88±6.59(P>0.05)。治疗4个月后HAMD评分10.76±5.32、明显低于对照组14.54±5.12(P<0.01);临床疗效(痊愈率44.44%、显效率22.22%、有效率33.34%)明显高于对照组(21.74%、21.74%、56.52%)(P<0.05);复饮率(24.44%)低于对照对照组(45.56%)(P<0.05)。结论认知疗法合并帕罗西汀治疗酒依赖伴发抑郁疗效较好,戒酒效果较好。     

CF-1小鼠胚胎成纤维细胞饲养层的制备**

背景：小鼠胚胎成纤维细胞作为干细胞生长用饲养层,优于无饲养层,它能够分泌一些既促进干细胞生长又能抑制干细胞分化的因子。 目的：建立CF-1小鼠胚胎成纤维细胞饲养层最佳的分离培养方法,分析丝裂霉素C抑制成纤维细胞增殖的最佳浓度与时间,用于培养诱导多能性干细胞。 方法：取孕10~15 d CF-1小鼠,分离胎鼠原代成纤维细胞,经不同质量浓度(5,10,15,20 mg/L)丝裂霉素C处理不同时间(1,1.5,2,2.5,3 h)制备饲养层,并观察其增殖情况。将人诱导多潜能干细胞或胚胎干细胞在丝裂霉素C处理过的饲养层上培养,观察细胞集落生长情况。 结果与结论：制备CF-1小鼠胚胎成纤维细胞饲养层的最佳胎龄为13.0~14.0 d。丝裂霉素C抑制CF-1小鼠胚胎成纤维细胞增殖的最佳浓度及作用时间为10 mg/L,2.5 h,成纤维细胞饲养层可以维持5~7 d。诱导多能性干细胞与胚胎干细胞在经10 mg/L丝裂霉素C处理2.5 h后饲养层上能发育成典型的“鸟巢”状干细胞集落。 关键词：胚胎成纤维细胞;诱导多潜能分化干细胞;饲养层;丝裂霉素C;细胞集落 doi:10.3969/j.issn.1673-8225.2012.10.017     

piggyBac转座子调节pPiggyBac-Rb质粒的构建*○

背景：外源性Rb基因的导入是抑制视网膜母细胞瘤细胞增殖的有效途径,piggyBac转座子介导的外源基因表达载体高效安全,在基因治疗方面具有广阔的应用前景。 目的：探讨piggyBac转座子介导的外源性Rb基因的转染效率及其对视网膜母细胞瘤细胞的抑制作用。 方法：采用RT-PCR技术扩增Rb基因编码区Rb cDNA,定向插入到载体pPiggyBac,获得PiggyBac调节的Rb表达质粒pPiggyBac-Rb;通过单独转染或合并pPiggyBac-helper转染人视网膜母细胞瘤细胞SO-RB50。 结果与结论：考马斯亮蓝染色证明piggyBac转座子系统的转染效率最高,荧光定量PCR以及免疫荧光实验证明其介导的目的基因Rb与宿主SO-RB50细胞基因组整合效率最高且可在细胞中长期稳定表达,MTT实验证明经其所介导的外源性Rb基因表达对细胞活性影响最为显著。结果提示piggyBac转座子有可能成为视网膜母细胞瘤基因治疗的安全有效载体。关键词：piggyBac转座子;SO-RB50细胞;转染;Rb基因;基因治疗 doi:10.3969/j.issn.1673-8225.2012.20.033     

腺病毒介导的骨形态发生蛋白基因增强在骨组织工程中的应用

背景：将目的基因转导到靶细胞中需要载体工具,载体可协助基因进入细胞及表达蛋白,载体的选择是转基因成败的关键。 目的：总结腺病毒介导的骨形态发生蛋白基因增强用于诱导成骨和修复骨缺损的研究现状。 方法：应用计算机检索PubMed 数据库及中国期刊网全文数据库1989-01/2012-01 有关腺病毒介导骨形态发生蛋白基因转染的诱导成骨作用,骨形态发生蛋白基因修饰的组织工程化骨修复骨缺损的文章,英文检索词为“adenovirus,bone morphogenetic protein, gene transfection, bone tissue engineering”,中文检索词为“腺病毒,骨形态发生蛋白,基因转染,骨组织工程”。排除重复性及非中英文语种研究,共保留24篇文献进行综述。 结果与结论：腺病毒介导的骨形态发生蛋白基因增强的组织工程利用基因工程技术将编码特定功能因子的基因转移到种子细胞上,使其持续产生生长因子。转基因后的细胞停留支架材料表面缓慢释放生长因子,促进成骨前体细胞增殖分化,也可在骨缺损处诱导成骨前体细胞向成骨细胞定向分化,从而修复骨缺损。      

磷酸钙骨水泥结合自体带血运骨膜修复节段性骨缺损

背景：磷酸钙骨水泥是一种新型骨替代材料,但具有降解慢、力学性能差等缺点。 目的：探讨磷酸钙骨水泥结合自体带血运骨膜修复节段性骨缺损的可行性。 方法：将72只新西兰大白兔随机数字法均分为4组：保留/不保留骨膜的不用磷酸钙骨水泥修复组、保留/不保留骨膜的磷酸钙骨水泥修复组。修复后4,12,24周取标本进行大体、放射学、组织学及生物力学测试。 结果与结论：不用磷酸钙骨水泥修复组：不保留骨膜12周时才开始出现骨痂,24周时骨端硬化,髓腔闭合;保留骨膜后4周时出现少量的骨痂,12周时骨痂增多,24周时形成新生骨。磷酸钙骨水泥修复组：不保留骨膜时4周材料与骨端分界明显,12周时分解模糊,24周时材料与骨端形成骨性连接;保留骨膜4周材料未见降解,12周时材料有降解,24周时材料吸收更多,髓腔再通。24周时磷酸钙骨水泥修复组行三点弯曲生物力学测试,不保留骨膜组最大负荷低于保留骨膜组(P < 0.05)。说明磷酸钙骨水泥较好促进骨的生长,自体带血运骨膜的存在有利于磷酸钙的吸收,促进骨的形成。     

应激性溃疡研究进展

应激性溃疡(stress ulcer, SU)是指机体在各类严重创伤、危重疾病等严重应激状态下,发生的急性消化道糜烂、溃疡等病变,最后可导致消化道出血、穿孔,并使原有病变恶化的急性致死性并发症.SU为应激引起的胃十二指肠粘膜急性病变,包括应激性溃疡及出血较合理的命名应为上消化道急性应激性粘膜病变(acute stress mucosal lesion ASML)[1].危重病人合并应激性溃疡大出血的死亡率目前仍在50%以上[2].而近期研究发现,高血压性脑出血急性期并发应激性溃疡出血高达14%～76%,致使其病死率高达78. 6%[3].因而应激性溃疡的发病机制及防治日趋被广大学者所重视.现就应激性溃疡的相关因素、发病机制、及预防措施进行综述.     

Establishment and Characterization of a Cell Based Artificial Antigen-Presenting Cell for Expansion and Activation of CD8+ T Cells Ex Vivo

Atificial antigen-presenting cells are expected to stimulate the expansion and acquisition of optimal therapeutic features of T cells before infusion. Here CD32 that binds to a crystallizable fragment of IgG monoclonal antibody was genetically expressed on human K562 leukemia cells to provide a ligand for T-cell receptor. CD86 and 4-1BBL, which are ligands of CO. stimulating receptors of CD28 and 4-1BB. respectively, were also expressed on K562 cells. Then we accomplished the artificial antigen-presenting cells by coupling K32, CD86/4-IBBL cell with OKT3 monoclonal antibody against CD3.named K32/CD86/4-lBBL/OKT3 cells. These artificial modified cells had the abilities of inducing CD8+ T cell activation. promoting CD8+ T cell proliferation, division, and long-term growth, inhibiting CD8+ T cell apoptosis, and enhancing CD8+ T cell secretion of IFN-Y and perforin. Furthermore, antigen. secific cytotoxic T lymphocytes could be retained in the culture stimulated with K32/CD86/4-1BBL/OKT3 cells at least within 28 day This approach was robust, simple, reproducible and economical for expansion and activation of CD8+ T cells and may have important therapeutic implications for adoptive immunotherapy. Cellular & Molecular Immunology.2007;5(1):47-53.     

中小学教师心理控制源、应付方式与自我和谐的关系

目的考察中小学教师自我和谐、心理控制源、应付方式的状况及其关系。方法使用自我和谐量表、内控性、有势力的他人及机遇量表、应付方式问卷对200名中小学教师进行调查。结果①教师整体上倾向内控,使用最多的应付方式是解决问题,最少的是自责;②任教地点、职务、职称、学历不同的教师自我和谐、应付方式有显著差异,年龄、教龄、任班主任年限、自评心理健康水平、生活压力、工作满意度与心理控制源、应付方式、自我和谐的不同维度相关显著;③机遇和幻想对自我和谐有正向预测作用,内控、解决问题、求助对自我和谐有负向预测作用。结论自我和谐可以作为衡量心理健康水平的一个标志,中小学教师心理控制源与应付方式对自我和谐有重要影响。     

Classification of duck hepatitis virus into three genotypes based on molecular evolutionary analysis

The nucleotide sequences of the complete VP1, VP0, VP3, and partial 3D regions of seven duck hepatitis virus (DHV) serotype 1 (DHV-1) strains isolated in China between 2001 and 2007 and one DHV-1 strain originally obtained from ATCC were determined and compared with previously available DHV sequences in GenBank. Phylogenetic analysis on the basis of VP1 sequences demonstrated three distinct genetic groups. There was an excellent concordance among the genetic groups assigned based on the complete VP0, VP3, and the partial 3D regions. In view of the growing importance of molecular techniques in diagnosis, we propose that the three genetic groups should be termed DHV types A, B, and C. All DHV-1 strains grouped in genotype A, whereas the new serotype strains isolated in Taiwan and the new serotype strains isolated in South Korea clustered into genotypes B and C, respectively, suggesting a potential genetic correlates of serotype. In pairwise comparisons of complete VP1, VP0, and VP3 nucleotide and amino acid sequences and the partial 3D nucleotide sequence, DHVs of the same genotype were clearly distinguished from those of heterologous genotypes. Analysis of the amino acid sequences of the three capsid proteins demonstrated the presence of conserved elements that form the eight-stranded beta-barrel structures, as well as intervening domains that vary in sequence between strains of different genotypes as seen in other picorna viruses.