人端粒酶逆转录酶启动子介导的靶向性肿瘤基因治疗的研究进展

端粒酶与肿瘤发生发展的关系是近年来肿瘤研究领域的热点之一．端粒的磨耗限制大多数体细胞的复制,肿瘤细胞主要通过转录上调端粒酶限制成分催化亚基端粒酶逆转录酶（hTERT）维持端粒长度．由于hTERT启动子区域已被克隆,其特点也已被鉴定,hTERT启动子介导的靶向性肿瘤基因治疗成为研究的热点．我们综述了hTERT启动子介导的靶向性肿瘤基因治疗最新研究进展．     

A fuzzy logic model for differential diagnosis of lower urinary tract dysfunctions

Lower urinary tract dysfunctions (LUTD) restrict quality of life, resulting in decreased work productivity and emotional well‐being. However, most people are not diagnosed because they do not seek medical treatment. In addition, some facilities do not adequately train health professionals in the evaluation, diagnosis and treatment of these conditions. The study's objective was to develop a decision support system modelled on fuzzy logic that defines LUTD using the terminology of the International Continence Society. This methodological study aimed to develop a model that uses the maximum–minimum composition (max–min) of fuzzy relations that can perform differential diagnoses of LUTD. The model was tested in 100 cases (50 men and 50 women), and the data were obtained from medical records containing the clinical data and results of urodynamic studies. All medical records were reviewed by a specialist in urology. The model was capable of determining a diagnosis in full (62%) or partial (36%) agreement with the medical report. Agreement between the model and the medical report was excellent (kappa = 0·98, p ? 0·001, CI = 0·88–1) or substantial (kappa = 0·53, p ? 0·001, CI = 0·45–0·60), considering overestimative accordance (where accordance is assumed when at least one diagnosis is equal) and underestimative accordance (where accordance is assumed when all diagnoses are equal), respectively. The proposed model based on the max–min composition of fuzzy relationships is very simple and performed well. However, more tests are recommended before the model is used as a decision support system.     

Effects of collagen membranes enriched with in vitro-differentiated N1E-115 cells on rat sciatic nerve regeneration after end-to-end repair

Preventing falls and fall-related fractures in the elderly is an objective yet to be reached. There is increasing evidence that a supplementation of vitamin D and/or of calcium may reduce the fall and fracture rates. A vitamin D-calcium supplement appears to have a high potential due to its simple application and its low cost. However, published studies have shown conflicting results as some studies failed to show any effect, while others reported a significant decrease of falls and fractures. Through a 15-year literature overview, and after a brief reminder on mechanism of falls in older adults, we reported evidences for a vitamin D action on postural adaptations - i.e., muscles and central nervous system - which may explain the decreased fall and bone fracture rates and we underlined the reasons for differences and controversies between published data. Vitamin D supplementation should thus be integrated into primary and secondary fall prevention strategies in older adults.     

体外原代培养纯化新生鼠嗅鞘细胞的实验方法：差速贴壁＋阿糖胞苷＋胰酶法

目的:通过细胞形态学观察及生物学特性鉴定,建立一种经济实用的体外原代培养纯化新生鼠嗅鞘细胞的实验方法。方法:实验于2006-06在武汉理工大学完成。①阿糖胞苷(Sigma,批号w10562);胎牛血清(杭州四季青产品);胰蛋白酶(Amresco);多聚赖氨酸(Sigma);胶质纤维酸性蛋白,神经生长因子受体蛋白抗体(博士德)。②选取出生3d内的Wistar大鼠5只,乙醇浸泡后断头处死,取嗅球的最外两层,通过1.25g/L胰蛋白酶消化分离嗅鞘细胞,体外原代培养。③采用Nash差速贴壁 阿糖胞苷 胰酶法纯化嗅鞘细胞。培养18h后将未贴壁的细胞悬液转种于另一未涂层的器皿中,再培养36h,重复上述步骤移入0.1g/L多聚赖氨酸包被的塑料培养瓶中进行培养,纯化后的细胞在24h内陆续贴壁,常规培养2d,加入终浓度为2mg/L的阿糖胞苷,作用48h后,换上新的培养基,继续培养1d,弃去培养基,用无钙镁的Hanks液清洗2次,然后用1.25g/L的胰酶消化10min,待细胞突起回缩、胞体变圆时,立刻加入纯血清终止,其血清终浓度为20%,制备单细胞悬液。④倒置显微镜下观察其形态变化,苏木精-伊红染色,同时行神经生长因子受体蛋白和胶质纤维酸性蛋白免疫组化染色鉴定嗅鞘细胞,并计算嗅鞘细胞阳性百分率。结果:①活细胞形态观察:分离培养的嗅鞘细胞具有双极或多极突起,且突起细长,相互交织。②嗅鞘细胞的苏木精-伊红染色鉴定:细胞呈三角形或梭形,有长的突起,胞浆被染成粉红色,胞核呈蓝紫色,胞核内深染的为核仁,有1～3个核仁。③嗅鞘细胞的胶质纤维酸性蛋白免疫组化染色鉴定:细胞呈现棕黄色,胞核淡染,阳性细胞成网状连接,胞体多为三角形,有细长突起,阳性率91.5%。④嗅鞘细胞的P75免疫组化染色鉴定:阳性细胞呈绿色,多数细胞染色阳性,阳性率89%。结论:实验所采用的Nash差速贴壁 阿糖胞苷 胰酶法分离纯化培养嗅鞘细胞切实可行,为神经诱导修复材料的研究提供种子细胞。     

认知矫正治疗对慢性精神分裂症患者临床症状和社会功能的影响

目的:观察认知矫正治疗对慢性精神分裂症患者临床症状和社会功能的改善作用。方法:选择2003-01/08在北京回龙观医院住院的慢性精神分裂症患者104例。均符合CCMD-Ⅲ及DSM-Ⅳ关于精神分裂症诊断标准;年龄25～55岁;病程≥2年;病情稳定,处于迁延、残留或部分缓解状态;药物治疗状况稳定,近期无换药打算;纳入对象或家属同意入组并签署知情同意书。应用随机数字表法将患者分认知矫正治疗组和对照组,每组52例。在相近药物治疗的基础上,认知矫正治疗组以Ann Delahunty和Rodney Morice等制定的神经认知矫正手册(汉化)为治疗工具,在治疗师的指导下进行认知作业练习,内容包括认知灵活性、工作记忆、计划执行功能3大功能模块。对照组予以相同时间的工娱治疗,主要包括有治疗师指导的操作性音乐治疗和舞蹈治疗。治疗前后两组患者分别进行PANSS、住院精神患者社会功能缺陷量表和护士观察量表的评定。结果:实验共纳入慢性精神分裂症患者104例,认知矫正治疗组44例,对照组46例进入结果分析,14例脱落。①治疗前后两组患者PANSS量表总分以及阴性症状量表、复合量表、一般精神病理量表、反应缺乏量表4个分量表的评分均有下降,组内比较差异有显著性意义(t=2.12～4.59,P<0.05);减分情况在两组间差异不明显(P>0.05)。②两组患者的社会功能缺陷量表总分在治疗后均有下降,与治疗前比较,差异有显著性意义(t=3.89,2.04,P<0.05);两组间比较,差异无显著性意义(P>0.05)。认知矫正治疗组治疗后护士观察量表的总病情以及总消极、迟滞2个分量表评分下降,与治疗前比较差异有显著性意义(t=1.49,1.19,2.81,P<0.05);其中迟滞项的减分在两组间比较,差异具有显著性意义(F=4.97,P<0.05)。③社会功能量表的改善与词语流畅性的改善呈正相关(R2=0.36,P<0.05),护士观察量表中总病情与积极两项评分的改善也与言语流畅性测验的改善正相关(R2=0.37,0.34,P<0.05)。结论:认知矫正治疗能在一定程度上改善精神分裂症患者的社会功能,并与部分认知功能的改善相关,但对临床症状无明显改善作用。     

Effect of endothelium on the actions of sympathetic and sensory nerves in the perfused rat mesentery.

We and others have previously demonstrated that pretreatment with capsaicin produces an augmentation of vasoconstrictor responses to transmural nerve stimulation. In the present study, removal of endothelium by saponin or inhibition of nitric oxide synthesis by N omega-nitro-L-arginine methyl ester produced an augmentation of vasoconstrictor responses to transmural nerve stimulation, responses which were further potentiated after treatment with capsaicin to desensitize sensory nerves. Capsaicin treatment decreased vasodilator responses to acetylcholine, but only at low acetylcholine concentrations. Potentiation by capsaicin of vasoconstrictor responses to transmural nerve stimulation was not affected by indomethacin. In the presence of guanethidine and methoxamine, transmural nerve stimulation caused vasodilator responses in the perfused rat mesentery. These responses were unaffected by removal of endothelium, as were vasodilator responses to exogenous calcitonin gene-related peptide (CGRP). In contrast, substance P did not produce any relaxation in the methoxamine-contracted mesentery. This study suggests that facilitation of vasoconstrictor responses to transmural nerve stimulation after capsaicin treatment primarily reflects inhibition of sensory nerve effects resulting in an increase of sympathetic vasoconstrictor actions. The present results also suggest that vasodilator responses to sensory nerve activation or exogenous CGRP are endothelium-independent and that substance P does not significantly contribute to modulation of vascular tone in the rat mesentery.     

Adrenergic nerves compensate for a decline in calcium buffering during ageing

1. The ubiquitous involvement of intracellular calcium ([Ca2+]i) in multiple neuronal pathways has led investigators to suggest that dysfunction of calcium homeostasis may be the primary mediator of age-related neuronal degeneration. Recently, it was shown that sympathetic neurones from superior cervical ganglion (SCG) of aged rats demonstrate decreased sarco-/endoplasmic reticulum Ca2+-ATPase (SERCA) function and that aged neurones are more dependent upon mitochondria to control K+-evoked [Ca2+]i transients. 2. Therefore, in the present study we investigated age-related changes in ATP-dependent calcium pumps of plasma membrane Ca2+-ATPase (PMCA) and SERCA in acutely dissociated SCG cells from Fischer-344 rats aged 6 and 20 months. To distinguish between PMCA and SERCA pump activity, we applied the Ca2+-ATPase blocker vanadate and measured rates of recovery of K+-evoked [Ca2+]i transients by fura-2 microfluorometry. 3. Young SCG cells showed a biphasic response to vanadate over the vanadate concentration range (0.01-100 microM); however, old SCG cells showed only a single response over the same concentration range. Additionally, old SCG cells showed a greater sensitivity to Ca2+-ATPase blockade by vanadate. 4. The contribution of mitochondrial calcium uptake to regulate [Ca2+]i was also investigated. To measure the impact of mitochondrial calcium uptake, PMCAs and SERCAs were blocked with vanadate (100 microM) and extracellular sodium was replaced with tetraethylammonium (TEA) to block Na+/Ca2+-exchange. Treated SCG cells showed a decline of 50% in rate of recovery of [Ca2+]i in both 6- and 20-month-old cells; however, this effect did not vary with age. 5. These data suggest that there is an age-related decline in function of SERCAs, with an increased reliance on PMCAs to control high K+-evoked [Ca2+]i transients. In addition, there appears to be no age-related change in the capacity of the mitochondria to restore [Ca2+]i transients to basal levels.