细胞因子诱导的自然杀伤细胞培养体系的评价

目的:评价采用γ-干扰素(IFN-7)、白介素-2(IL-2)、抗 CD3 单抗 3 因子对外周血来源的 CIK 细胞培养的效果.方珐:采集17例实体肿瘤患者的外周血,分离单个核细胞培养,于培养第 1 天加入重组人IFN-Y,第2天加入抗人 CD3 单抗、重组人IL-2,隔天半量换培养液并添加人重组IL-2以维持其浓度...     

蒙药“通拉嘎-5”对脂肪细胞葡萄糖摄取的影响及其机制研究

目的探讨"通拉嘎-5"对3T3-L1脂肪细胞葡萄糖摄取的影响及其作用机制。方法将3T3-L1前脂肪细胞诱导分化为脂肪细胞后,"通拉嘎-5"作用于3T3-L1脂肪细胞24 h,用3H标记的2-脱氧-葡萄糖([3H]2-DG)示踪检测胰岛素介导的3T3-L1脂肪细胞摄取葡萄糖的情况;"通拉嘎-5"作用3T3-L1脂肪细胞48 h后,用Real-time PCR检测PPARγ和LXRα的基因表达,用Western blot检测PPARγ和LXRα的蛋白表达。结果 "通拉嘎-5"能促进胰岛素介导的3T3-L1脂肪细胞的葡萄糖摄取(比阴性对照组高1.915倍,P<0.05);能明显上调3T3-L1脂肪细胞和LXRα基因(比阴性对照组高2.895倍,P<0.01)和蛋白(比阴性对照组高1.977倍,P<0.01)表达;而对PPARγ基因和蛋白表达无明显的影响。结论 "通拉嘎-5"可能通过提高3T3-L1脂肪细胞LXRα的基因和蛋白表达,促进胰岛素介导的葡萄糖摄取功能,改善胰岛素抵抗,可能为"通拉嘎-5"治疗非酒精性脂肪肝(NAFLD)的作用机制之一。     

Troglitazone Induced Apoptosis via PPARγ Activated POX-induced ROS Formation in HT29 Cells

Objective In order to investigate the potential mechanisms in troglitazone-induced apoptosis in HT29 cells,the effects of PPARγ and POX-induced ROS were explored.Methods [3-(4,5)-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay,Annexin V and PI staining using FACS,plasmid transfection,ROS formation detected by DCFH staining,RNA interference,RT-PCR & RT-QPCR,and Western blotting analyses were employed to investigate the apoptotic effect of troglitazone and the potential role of PPARγ pathway...     

北五味子配方颗粒提取工艺的正交设计试验优选

目的：优选北五味子配方颗粒的最佳提取工艺。方法：采用正交设计安排试验,以提取率、五味子甲素（DSSD）、五味子乙素（γ－SSD）的含量作为考察指标,考察用水量、提取时间、提取次数对五味子配方颗粒提取工艺条件的影响,优选出合理的提取工艺。结果：在既定的HPLC测定方法下,北五味子配方颗粒中DSSD及γ-SSD含量分别在0．265。1．590珥g和0．638～3．188μg范围内,进样量与峰面积的线性关系良好（r=1．0000和r=0．9999）,平均回收率分别为101．12％和100．38％,RSD值分别为1．75％和1．10％。影响北五味子配方颗粒提取率、DSSD、γ-SSD含量的主要因素依次为提取次数、提取时间、用水量,最佳工艺条件为：加水量为10倍,提取3次,每次2h,测得DSSD平均含量为0．024％,γ-SSD平均含量为0．049％,平均提取率为36．12％。结论：该提取工艺具有提取率稳定性好、五味子甲素及五味子乙素得率高、操作简单且成本较低的特点,可为五味子配方颗粒的提取工艺提供参考。     

Increased ratio of calcineurin immunoreactive neurons in the caudate nucleus of patients with schizophrenia

Calcineurin (CaN) has been investigated extensively in numerous biochemical, behavioral, and genetic studies in schizophrenia because its function is closely related to dopamine-glutamate signal transduction, which is thought to be associated with pathophysiological changes in schizophrenia. Although evidence has suggested that dysfunction of CaN may be a risk factor for schizophrenia, there have been few reports focusing on the expression of CaN mRNA and CaN protein levels in the brains of schizophrenic patients. In addition, findings on CaN expression in postmortem brains from patients with schizophrenia have been inconsistent. Here, we conducted immunohistochemical examinations of several regions in postmortem brains, including the dorsolateral prefrontal cortex (DLPFC), hippocampus, caudate nucleus, and putamen, using specific antibodies, and compared the results from the brains of nine schizophrenic subjects to nine age- and sex-matched control subjects. There was no significant difference in the ratio of CaN immunoreactive (IR) neurons between schizophrenia and control groups in the DLPFC or hippocampus, and a significantly increased ratio of CaN-IR neurons was seen in the caudate nucleus in the brains from schizophrenia patients. As the striatum contains most of the brain dopamine, the results of the present study have critical implications and suggest that alterations in CaN signaling in the caudate contribute to the pathogenesis of schizophrenia. This is the first report of caudate CaN abnormalities in schizophrenia.     

Adjuvant interferon-γ treatment in two cases of refractory tuberculosis of the brain

Tuberculosis (TB) of the brain is often refractory and has the highest morbidity and mortality among the mycobacterial infections. A recent report suggests that interferon-γ may be of help since it can modulate the host inflammatory response against mycobacteria in the brain. Here, we report on a 44-year-old woman with multiple tuberculomas in the brainstem and a 40-year-old man with two large TB abscesses in the brain, both of whom had no response to anti-TB medications for 5 and 7 months, respectively, but with near-complete resolution with adjuvant interferon-γ therapy (50 μg/m(2), subcutaneously, three times per week). Our cases show that refractory brain TB in immunocompetent patients can be successfully treated with adjuvant interferon-γ therapy, without any significant side effects.     

Induction of interferon-γ contributes to Toll-like receptor 3-mediated herpes simplex virus type 1 inhibition in astrocytes

Toll-like receptor 3 (TLR3) recognizes double-stranded RNA and induces type I interferon (IFN)-mediated antiviral immunity against a number of viral infections. Type III IFN (IFN-λ) is a newly identified antiviral cytokine that has biological functions similar to those of type I IFNs. We thus investigated the role of IFN-λ in TLR3 activation-mediated inhibition of herpes simplex virus type 1 (HSV-1) in human primary astrocytes. Human astrocytes express endogenous IFN-λ1 and IFN-λ receptor complex, interleukin-28 receptor α subunit (IL-28Rα), and IL-10Rβ. The activation of TLR3 by poly-I:C treatment significantly induced the expression of IFN-λ1 and IFN-λ2/3 in astrocytes. The induction of IFN-λ contributed to TLR3 activation-mediated HSV-1 inhibition in astrocytes. Investigation of the mechanisms showed that treatment of astrocytes with specific antibody against IFN-λ receptor attenuated the anti-HSV-1 activity of poly-I:C, indicating that endogenous IFN-λ contributes to the anti-HSV-1 effect of TLR3 activation. The anti-HSV-1 effect of endogenous IFN-λ was also confirmed by the finding that recombinant IFN-λ treatment inhibited HSV-1 infection of astrocytes. These results provide direct and compelling evidence that endogenous IFN-λ participates in TLR3-mediated antiviral activity, which may have important implications in host cell innate immunity against HSV-1 infection in the CNS.     

225名处级干部部分生化体检指标检测结果分析

目的 探讨邯郸矿业集团225名矿处级体检干部中,脂肪肝组与非脂肪肝组血液部分生化检测指标间的关系,提醒其关注自身健康.方法 对邯郸矿业集团总医院2010年7～11月体检中心接诊的225名矿处级干部的三酰甘油(TG)、空腹血糖(FPG)、γ-谷氨酰转移酶(γ-GGT)、血尿酸(UA)检测指标进行回顾性调查分析,并按其是否患有脂肪肝将结果分为两组,并进行统计学分析研究.结果 脂肪肝组与非脂肪肝组的TG、FPG、γ-GGT、UA异常检出率分别是59.84%、41.73%、74.02%、15.75%和24.49%、18.37%、18.37%、1.02%,脂肪肝组各项指标异常检出率均明显高于非脂肪肝组,两者差异有统计学意义(P<0.01).并且两组TG、FPG、γ-GGT、UA的平均浓度,脂肪肝组明显高于非脂肪肝组,两者差异具有统计学意义(P<0.01).结论 高血脂、高血糖是诱发脂肪肝的重要因素,脂肪肝患者是发生肝病、痛风和心脑血管病的高发人群,领导干部健康问题不容忽视,应加强锻炼,合理膳食,积极治疗,提高身体健康水平.     

氮氧自由基R-1对人肝细胞L-02的辐射防护作用

目的 探讨氮氧自由基R-1(简称R-1)对人肝细胞L-02的辐射防护作用.方法 在L-02细胞培养液中,加入终浓度为0、0.125、0.25、0.5、1、2、4、8、16、32μmol/L的R-1,作用24、48和72 h.用MTT法测定R-1的毒性作用,以筛选合适的R-1浓度.后续实验选择0.125、0.25、0.5、1 μmol/L 4个浓度组检测其防护作用.设终浓度4 mmol/L的细胞保护剂WR2721为阳性对照组.采用60Coγ射线照射,吸收剂量为0、1、2、4、8 Gy,照射72 h后,进行MTT比色法.L-02细胞分为2组:照射前30 min加药组和照射后立即加药组.终浓度均为0.25 μmol/L,吸收剂量为4 Gy,照射后72 h进行MTT比色实验.照射后10 d,用克隆形成实验检测不同浓度的R-1对L-02细胞活力的影响.选用防护效果最佳的0.25μmol/L浓度对细胞进行预处理,分别在4 Gy照射后的24、48和72 h,用倒置显微镜观察细胞形态的变化,Hoechst 33258荧光染色法和流式细胞仪检测细胞凋亡.结果 当R-1浓度低于1 μmoL/L时,与0 μmol/L组相比,L-02细胞各时间点的吸光度(A)值无明显变化;而浓度高于2 μmol/L时,与0 μmol/L组相比,其A值随浓度的增高而下降.选用0、0.125、0.25、0.5、1μmol/L的浓度组.与照射组相比,R-1各浓度组的A值和克隆形成率明显提高,其中0.25μmol/L组的作用最明显.与照射组相比,0.25 μmol/L预处理组的L-02细胞贴壁好,折光性强,轮廓清晰,凋亡细胞和死亡细胞明显较少.结论 R-1能有效地防护60Coγ射线对L-02细胞的辐射损伤,其防护作用可能与减少细胞凋亡有关.

Abstract：

Objcetive To investigate the protective effects of the nitroxides R-1 on human liver cells exposed to ionizing radiation.Methods Human liver cells L-02 were cultured and irradiated with 60Co γ-rays at the doses of 0,1,2,4,and 8 Gy,in order to screen the proper irradiation dose.WR2721 at the terminal concentration of 4 mmol/L was used as positive control.L-02 cells irradiated with 4 Gy were added with R-1 at the terminal concentration of 0.25 μmol/L at 30 min before irradiation or immediately after irradiation.MIT method was used to screen the proper conditions for follow-up experiment 72 h later.L-02 cell culture fluid was added with R-1 at the concentrations of 0,0.125,0.25,0.5,and 1 μmol/L,respectively for 30 min before irradiation at the doses of 0,1,2,4,and 8 Gy to ealculate clone formation rate at 10 d post-irradiation.L-02 cells were cultured and divided into 4 groups:control group without any treatment.drug group pretreated by 0.25 μmol/L R-1 only,irradiation group,irradiated at 4 Gy only,and drug + irradiation group with combination of 0.25 μmol/L R-01 and 4 Gy irradiation.The inverted microscopy and Hoechst 33258 staining and flow eytometry were used to observe the apoptosis of the cells at 24,48,and 72 h later.Results Nitroxides R-1 did not inhibit the viability of L-02 cell when its concentration was less than 1 μmol/L and it inhibited the L-02 cell growth when the concentration wu higher than 2 μmoL/L.The A value and colony formation rate of different concentration of R-1 groups were all higher than those of the irradiation group,and the effect of the 0.25 μmol/L drug concentration group was the most significant.Consequently,the concentration 0.25 μmoL/L was selected for follow-up experiment.Compared with the irradiation group,the L-02 cells of the pretreatment group showed solid adherence, increased refraction,clear outline,less apoptotic and dead cells at 4 Gy post-irradiation.Conclusions Nitroxides R-1 can protect the human liver cells from 60Coγ-ray induced injury effectively.The mechanism of its protective effect may be the reduction of apoptosis.