DC2.4表面分子与RelB基因表达的关系

目的 探讨DC2.4表面分子表达水平与RelB基因表达间的关系.方法 用RPMI 1640完全培养液培养DC2.4细胞系,光学显微镜观察培养细胞的形态特征;透射电镜观察DC2.4的细胞结构;流式细胞术分析DC2.4的表面标记(MHC-Ⅱ、CD86和CD40);RT-PCR检测DC2.4的RelB表达水平.结果 光镜观察DC2.4细胞表面有明显的树突状突起,形态极不规则;透射电镜显示DC2.4细胞内含许多质地均匀的脂滴,可见吞噬小泡样的结构.流式细胞术显示MHC-Ⅱ类分子和CD40呈低水平表达,而CD86分子却呈高水平表达;RT-PCR结果显示RelB呈低水平表达,与骨髓源性DC中的未成熟状态DC的RelB表达水平接近.结论 DC2.4具有强吞噬功能,其表面CD40分子和细胞内RelB基因均呈低水平表达,提示DC2.4是一种未成熟状态的细胞系.     

CD137信号对小鼠树突状细胞表面TLR4表达的调节

目的 探讨CD137（4-1BB）激发型单克隆抗体2A对小鼠未成熟树突状细胞株DC2.4表面TLR4表达的调节。方法 以流式细胞术检测不同剂量2A、LPS,或LPS预刺激后再加入2A等因素作用下,DC2.4表面的TLR4表达情况。结果 LPS下调TLR4表达,2A使DC2.4上TLR4表达上调,并拮抗LPS对TLR4的下调作用。LPS预处理后再加入2A,也拮抗LPS的下调作用。结论 CD137信号可以上调DC2.4表面TLR4的表达。     

反相高效液相色谱－紫外吸收法测定灵芝中氨基酸的含量

采用本实验室自行制备的反相高效液相色谱柱，以醋酸钠缓冲液和乙腈－水线性梯度洗脱，在室温下对常见的１６种１－氟－２，　４－二硝基苯（ＦＤＮＢ）衍生化氨基酸进行了有效的分离，用紫外吸收检测器（３６０ｎｍ）和外标定量法对灵芝蛋白中各种氨基酸的含量进行了测定。结果表明：ＦＤＮＢ衍生化氨基酸在一周内是稳定的，回收率在９６％～１０２％之间，相对标准偏差小于４％，灵芝中含有人体必需的赖氨酸、苯丙氨酸和组氨酸等组分。     

Foxp3,Galectin-9,PD-L1在小鼠DC2.4细胞的表达

目的观察CD80,CD86,Foxp3,Galectin-9,PD-L1在小鼠DC细胞的表达,了解小鼠DC2.4细胞系的免疫学特性。方法用含10%胎牛血清的RPMI-1640培养液培养小鼠DC2.4细胞,光学显微镜观察细胞形态;流式细胞术检测CD80,CD86,Foxp3,Galectin-9,PD-L1在DC2.4细胞表达。结果光学显微镜示小鼠DC2.4细胞贴壁生长,呈树突、梭形及不规则形;流式细胞术检测DC2.4细胞CD80表达为91.04%±4.90%,CD86表达为98.54%±1.96%,Foxp3无或微表达,Galectin-9表达为14.33%±2.94%,PD-L1表达为98.80%±0.63%。结论小鼠DC2.4细胞是一个高表达CD80,CD86及PD-L1,低表达Galectin-9,无或微表达Foxp3的细胞系。     

TLR8在小鼠DC2.4细胞中的表达及其作用

目的探讨应用CL075和Poly(dT)后,TLR8及其相关细胞因子IL-12、IL-10在小鼠DC2.4细胞中的表达情况。方法常规培养DC2.4细胞,光学显微镜观察未刺激组、Poly(dT)刺激组、CL075和Poly(dT)共同刺激组细胞的形态变化;Real-timePCR测定TLR8、IL-12、IL-10的mRNA表达;ELISA测定IL-12、IL-10的蛋白表达;Western blot测定TLR8的蛋白表达。结果未刺激及Poly(dT)刺激组细胞较幼稚,树突较短且少。CL075和Poly(dT)刺激组细胞活化,树突增多且伸长;Poly(dT)刺激组DC2.4细胞中TLR8、IL-12、IL-10的mRNA及蛋白表达量与阴性对照组相比均无统计学意义。CL075和Poly(dT)刺激组DC2.4细胞中TLR8、IL-12的mRNA及蛋白表达量较阴性对照组明显升高(P<0.05或P<0.01),而IL-10的mRNA及蛋白表达量则明显低于阴性对照组(P<0.01或P<0.05)。结论应用CL075和Poly(dT)后,TLR8、IL-12在小鼠DC2.4细胞中的表达水平明显升高,而IL-10的表达水平则明显降低。     

Anti-morphine antibody contributes to the development of morphine tolerance in rats

Previous studies have shown that tolerance to the antinociceptive effect of morphine develops after a prolonged exposure, but its mechanisms remain unclear. In the present study, we examined whether anti-morphine antibody produced by chronic morphine exposure would contribute to the development of morphine antinociceptive tolerance in rats. Our results showed that anti-morphine antibody was present in rats rendered tolerance to antinociception after intrathecal morphine exposure for seven consecutive days. Superfusion of anti-morphine antibody onto spinal cord slice dose-dependently produced an inward excitatory current in spinal cord dorsal horn neurons using whole-cell patch-clamp recording, which surpassed morphine-induced outward inhibiting current. Co-administration of morphine with a monoclonal antibody (2.4G₂) against Fc receptors for seven days significantly attenuated the production of anti-morphine antibody as well as the behavioral manifestation of morphine tolerance in same rats. These results indicate that anti-morphine antibody produced by morphine exposure may contribute to the development of morphine tolerance possibly through counteracting the inhibitory morphine effect on spinal cord dorsal horn neurons.     

Photodynamic Therapy Using a New Photosensitizer Gallium Porphyrin Complex in the Treatment of Experimental Pancreatic Cancer

Abstract: To determine the therapeutic effect of photodynamic therapy (PDT) using a new photosensitizer 2.4-bis (1-decyloxyethyl)-Ca-deuteroporphyI diaspartic acid (Ga -DP), 20 female Syrian golden hamsters with experimentally induced pancreatic cancer were irradiated with a pulsed Nd : YAG laser with a Q switch after administration of Ga -DP. To induce pancreatic cancer, N-nitroso-bis-(2-oxypropyI) amine (BOP) at 10 mg/kg body weight was injected subcutaneously once a week for eight weeks. Ten of the twenty animals served as a control group. The other ten received laser treatment beginning 20 weeks after BOP administration. Twenty-four hours after intravenous injection of 3 mg/kg Ga-DP, pulsed Nd : YAG laser irradiation was performed using a Q switch with 1 W of power output for 3–5 minutes at a frequency of 300 Hz. Elimination or necrosis of the entire tumor resulted with tumors 6–10 mm in diameter in 6 animals. In 3 animals which had tumors more than 12 mm in diameter, cancer cells remained at the margins of the treated area. These results were felt to not only reflect the thermal effect of laser therapy, but also the effect of PDT combined with Ga-DP. They suggest that Ga-DP could become a clinically valuable photosensitizer in combination with PDT. (Dig Endosc 1994; 6 : 28–33)     

环丙沙星中间体,2,4—二氯—5—氟苯甲酰氯的合成

目的合成环丙沙星中间体２,４－二氯－５－氟苯甲酰氯。方法 以１,３－二氯－４－氟苯为原料,经取代,氧化,经三步反应制得。结果 反应条件温和,方法简单,总收率６９％。结论 适用于工业化生产。