广东省中山市头癣现状及其病原体分离结果分析

目的:了解广东省中山地区头癣现状及其病原体分布、流行情况。方法:对107例镜检阳性病发进行真菌培养、鉴定和临床分析。结果:107例患者中白癣81例(占75.7％),脓癣24例(22.4％),黑癣2例(1.9％),未发现黄癣。107例病发分离出5种共101个菌株,其中犬小孢子菌90株(占89.1％),须癣毛癣菌5株(4.9％),红色毛癣菌3株(2.79％),断发毛癣菌2株(1.98％),紫色毛癣菌1株(0.99％)。结论:广东省中山地区头癣种类以白癣为主,脓癣次之,黑癣少见,黄癣基本消失,流行的病原菌以亲动物性的犬小孢子菌占绝对多数。发病年龄以1岁～15岁的少年儿童为主,成年人少见。     

纤维蛋白胶体外重建兔角膜上皮的实验研究

目的利用组织工程技术体外重建角膜上皮组织,为重建角膜表面提供良好的移植材料和方法。方法以等量凝血酶和纤维蛋白原作用生成的纤维蛋白胶为载体(实验组),种植兔角膜缘组织块,体外培养重建角膜上皮层,每天记录细胞生长覆盖面积,从生长特性、光镜特征、超微结构特点以及免疫细胞化学方面进行观察检测。结果角膜缘上皮细胞在纤维蛋白胶上黏附生长增殖,体外培养7d左右即形成单层上皮细胞,2周左右形成的复层上皮细胞与纤维蛋白胶构成角膜上皮组织经检测与生理状态下的角膜上皮组织相近。实验组与对照组(置盖玻片为载体)比较细胞生长覆盖面积第4天至第7天差异有显著性(P<0.05)。结论以纤维蛋白胶为载体,体外培养的角膜上皮组织片可作为眼表重建角膜上皮移植良好的来源。     

Expression of human T-cell lymphotropic virus type-1 gene products in the short-term cultured skin tissues of an adult T-cell leukemia/lymphoma patient with cutaneous manifestations.

Adult T-cell leukemia/lymphoma (ATLL) is recognized as a disease etiologically associated with human T lymphotropic virus type-1 (HTLV-1) infection, but, neither viral replication nor specific virus antigen expression have been detected on ATLL cells distributed in organs, including skin. To examine the latent expression of HTLV-1 in the cutaneous lesions of ATLL patients, we cultured the lesional skin tissues in vitro and applied immunofluorescence staining with mouse monoclonal antibodies Lt-4, GIN-14, and F10, which react with p40tax, p19 and gp21, respectively. We recognized HTLV-1 specific antigens on clustered ATLL cells only in the deeper dermis of the skin after 24 hrs cultivation of the lesional skin tissue from an ATLL patient in RPMI-1640 medium supplemented with 20% fetal calf serum. In the electron microscope, we observed HTLV-1 like particles, 80-140 nm in diameter with envelope and core structures, in the same tissue specimen. These findings suggest that HTLV-1 gene products may be expressed in the skin lesions of ATLL patients and involved in the pathogenesis of skin eruptions in cutaneous type ATLLs. To our knowledge, this is the first report that envisages the potency of intracutaneous HTLV-1 expression in vivo.     

Patch clamp studies on the kinetics and selectivity of N-methyl-d-aspartate receptor antagonism by memantine (1-amino-3,5-dimethyladamantan)

Memantine (1-amino-3,5-dimethyladamantan) was tested as an antagonist of N-methyl-d-aspartate (NMDA) receptors on cultured superior collicular and hippocampal neurones using the patch clamp technique and its actions were compared to those of Mg²⁺ ions, ketamine, dextrorphan, dextromethorphan, phencyclidine and dizocilpine (MK-801). Memantine (2–33 μM) concentration-dependently antagonized responses to NMDA 100 μM with an IC₅₀ of 2.92 ± 0.05 μM. In contrast, current responses to (S)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (l-AMPA 50–100 μM) and γ-amino butyric acid (GABA 10 μM) were unaffected by Memantine 8 μM. Memantine 8 μM caused a non-parallel shift of the NMDA concentration-response curve to the right in a manner indicative of uncompetitive open channel block. The effects of memantine were similar to ketamine in that both antagonists were weakly use- and strongly voltage-dependent. In contrast, MK-801, phencyclidine and dextrorphan showed much slower kinetics that was reflected in their marked use- and weaker voltage-dependency. The antagonistic effects of memantine were not reversed by increasing concentrations of glycine (0.1–100 μM) ruling out the possibility of an interaction of memantine with the strychnine-insensitive glycine modulatory site associated with the NMDA receptor-channel complex. Memantine (1–100 μM) also selectively antagonized responses to NMDA (40 μM) in the cortical wedge preparation with IC₅₀ of 12.9 ± 1.5 μM.     

序贯培养对小白鼠胚胎体外发育的影响

目的:探索序贯培养技术对小白鼠胚胎体外发育的影响。方法:将 2 -细胞期的小鼠胚胎随机分为序贯培养组与传统培养组进行培养 ,观察分析两组的 8-细胞期胚胎形成率、桑椹期胚胎形成率及囊胚形成率。结果:序贯培养组 8-细胞期胚胎形成率、桑椹期胚胎形成率及囊胚形成率分别为 70 .5 %、6 8.2 %、6 5 .9% ,传统培养组分别为 4 5 .7%、4 5 .7%、34.3%。两组比较 ,序贯培养组高于传统培养组 ,差异有统计学意义 (P <0 .0 5 )。结论 :采用适当的培养液进行序贯培养 ,更适合小白鼠胚胎的体外发育     

人皮肤角质形成细胞的分离与原代无血清培养

目的 :建立一种体外人皮肤角质形成细胞分离与原代培养的方法。方法 :采用两步消化法对皮肤进行消化 ,获取角质形成细胞进行体外无血清培养基培养 ,进行细胞形态学观察、免疫组织化学、透射电镜鉴定及生长曲线的绘制与分析。结果 :该方法可获取较多高纯度的角质形成细胞 ,且在体外可快速稳定增殖。结论 :两步消化法和体外无血清培养是一种理想的皮肤角质形成细胞分离培养的方法     

酸性环境介导皮质神经元损伤及阻断酸敏感离子通道1α的神经保护作用

目的 探讨脑梗死模型中酸性环境能否介导皮质神经元损伤及阻断酸敏感离子通道1α（acidsensing ion channels1α，ASIC1α）的神经保护作用。方法 制作缺氧细胞模型、脑梗死动物模型，检测ASIC1α阻断剂对乳酸脱氢酸（lactate dehydrogenase，LDH）释放率、大鼠行为学、脑梗死体积以及ASIC1蛋白表达的影响。结果 酸性环境可以损伤皮质神经元，加重缺氧对细胞的毒性作用，经侧脑室注射ASIC1α阻断剂可以改善脑梗死大鼠的行为学改变，梗死体积明显减小（P〈0.05）；随梗死时间延长，缺血半暗带ASIC1表达增加，在缺血后24h表达量较明显。结论 酸性环境导致皮质神经元损伤，其损伤机制与缺血后神经元ASIC1α开放以及表达增加有关；阻断ASIC1α具有神经保护作用。     

Oxidative modification of high density lipoprotein induced by cultured human arterial smooth muscle cells

Objective: To observe the oxidative modification of high density lipoprotein (HDL) induced by culturedhuman arterial smooth muscle cells (SMCs). Methods: HDL cocultured with SMCs at 37℃ in 48 h was subjected,and native HDL (N-HDL) served as control. Oxidative modification of HDL was identified by using agarose gel electro-phoresis. Absorbances of conjugated diene (CD) and lipid hydroperoxide (LOOH) were measured with ultravioletspectrophotometry at 234 and 560 nm respectively, and fluorescence intensity of thiobarbuturic acid reaction substance     

人外周血单核细胞和脐血CD34+细胞来源的树突细胞比较

目的比较从人外周血单核细胞及脐血CD34+细胞诱导树突细胞(DC)的方法及其特性.方法用淋巴细胞分离液分离外周血单个核细胞,贴壁法获得单核细胞;经GM-CSF及IL-4诱导获得DC.磁性活化细胞分选系统(MACS)分选脐血CD34+细胞,以GM-CSF、IL-4、TNF-α、SCF、FL、TPO等细胞因子诱导获得DC.用电镜、普通显微镜观察DC形态;流式细胞仪分析DC表型;同种混合淋巴细胞反应(allo-MLR)观察DC刺激T淋巴细胞增殖的能力.结果外周血单核细胞在因子GM-CSF、IL-4、TNF-α,脐血CD34+细胞在GM-CSF、IL-4、TNF-α、SCF、FL、TPO等因子作用下,可生成DC,并表达相应的DC分化抗原CD14、CD80、CD83、CD86、HLA-DR.脐血CD34+细胞具有较强的扩增能力,经2周诱导,体系细胞数可增加173.8士26.7倍,两种来源的DC均能刺激同种异体淋巴细胞的增殖反应.结论外周血单核细胞与脐血CD34+细胞,在体外均可诱导成DC,并具有相应的DC分化抗原和功能.