Investigation of gene–gene interactions between CD40 and CD40L in Polish multiple sclerosis patients

CD40–CD40L interaction is necessary for the activation of both humoral and cellular immune response and has been suggested to play a role in the pathogenesis of multiple sclerosis (MS). Therefore, we analyzed the combined influence of the CD40 and CD40L variants on MS susceptibility and progression on well-defined Polish population. Our investigation revealed that CT individuals in rs1883832 locus of CD40 possessed almost 1.5-fold higher risk for MS than CC individuals (OR = 1.44; 95%CI = 1.03–2.1; p = 0.032), while this risk for TT individuals was almost 2.5-fold higher (OR = 2.36; 95%CI = 1.19–4.78; p = 0.014).     

Inhibitors of TLR-4, NF-κB,and SAPK/JNK signaling reduce the toxic effect of lipopolysaccharide on RAW 264.7 cells

The present study was designed to examine and compare the effects of three suppressors on the cytokine response in tandem with examining: the synthesis of inducible forms of heat shock proteins; HSP72 and HSP90α; activities of NF-κB and SAPK/JNK signaling pathways; and TLR4 expression. Pre-treatment with inhibitors offers promise as protective means to lower the activity of these cascades, thereby circumventing the formation of excessive amounts of pro-inflammatory molecules. Three inhibitors of TLR4, SAPK/JNK, and NF-κB signaling, namely CLI-095, SP600125, and IKK Inhibitor XII, respectively, were added to cultured RAW 264.7 macrophages before the Escherichia coli lipopolysaccharide (LPS) application. Treatments of RAW 264.7 cells with each of the inhibitors resulted in a reduced response to LPS as was visualized by a decrease of TNF-α, IL-1, and IFN-γ production. In addition, inhibitors of the NF-κB and SAPK/JNK signaling reduced IL-6 production in LPS-treated cells, whereas the IKK inhibitor XII also decreased IL-10 production. Further, the NO production in LPS-stimulated macrophages was significantly reduced following application of CLI-095 or IKK inhibitor XII. The results also showed that the inhibitors suppressed TLR4 production and decreased phosphorylation of NF-κB and SAPK/JNK proteins, thereby preventing the activation NF-κB and SAPK/JNK signaling pathways in LPS-activated cells. In addition, the production of inducible heat shock proteins, HSP72 and HSP90-α, was reduced in LPS-stimulated RAW 264.7 cells pre-treated with inhibitors. These results suggest that inhibitors CLI-095, SP600125, and IKK inhibitor XII demonstrate potential effectiveness in the reduction of the inflammatory response by mechanisms involving both the cellular defense system and cellular signaling. In conclusion, suppressor of NF-κB cascade, IKK inhibitor XII, seems to be the most effective anti-toxic agent among studied inhibitors.     

PLAGL2对SP—C基因表达调控的作用靶点研究

目的 研究PLAGL2对SP—c基因表达调控的作用靶点。方法 定点诱变技术分别突变掉SP—C基因启动子上PLAGL2的结合位点以及PLAGL2的第6、7位锌指结构域序列并获得相应的突变体；凝胶电泳迁移实验研究PLAGL2及其突变体分别与SP—C基因启动子及其突变体之间的相互结合作用。结果 PLAGL2能与同位素标记的SP—C基因启动子片段相结合形成蛋白-DNA复合物，而两者的突变体却不能与相应的蛋白或DNA片段发生相互作用。结论 PLAGL2通过其第6、7位锌指结构与SP—C基因启动子上PLAGL2的核心和簇结合位点序列相互结合从而实现对SP—C基因的表达调控。     

Isolation of Side Population Cells and Detection of ABCG2 from SW480

Objective:Side population cells(SP cells)are a new type of stem cells.They mainly express ABCG2/BCRP1 and have the ability to eliminate DNA dye Hoechst33342.Many studies showed that side population cells were able of self-renewal,differentiation and carcinogenesis in cancers.Our investigation aimed at isolation of side population cells and ABCG2 positive subpopulation from colon cancer cell line SW480 and identification of their characteristics of cancer stem cells.Methods:side population cells and non-side population cells of colon cancer cell line SW480 were isolated with DNA dye Hoechst33342 and their cell cycles were measured by flow cytometry.Expression of ABCG2 of SW480 was measured by immunohistochemistry and immunofluorescence,and its proportion was measured by flow cytometry.Results:SW480 contained 2.29% side population cells.The fraction of side population cells decreased greatly to 0.40% by treatment with verapamil.The fraction of side population cells in S-G2M cell cycle was 16.14%,which was much lower than the fraction(34.05%)of non-side population cells in S-G2M.In SW480,ABCG2 positive cells,which proportion was 9.66%,were small,circular or oval,lack of psuedopods,similar to poor differentiation.On the contrary,the ABCG2 negative cells were large,polygonal,with many psuedopods,similar to high differentiation.Conclusion:our assay identified that side population cells did exist in SW480 and had a quiescence characteristic of stem cells.ABCG2 positive subpopulation occupied about 9.66% of SW480 and may have the ability to promote cell self-renewal and inhibit cell differentiation.Therefore,to isolate ABCG2 positive subpopulation from side population cells may be an alternative to study colorectal cancer stem cells.     

枳术丸煎剂与枳术汤对大鼠P物质的影响

目的探讨枳术丸与枳术汤对正常大鼠血浆P物质(SP)及肠组织SP的影响。方法正常大鼠分5组,灌胃给药7 d后采集标本,放免检测血浆及肠组织SP含量。结果枳术丸与枳术汤均能升高大鼠血浆SP及肠组织SP含量,呈一定的量-效关系,但枳术丸与枳术汤二者无显著差异。结论升高血及胃肠组织SP含量是枳术丸与枳术汤促进胃肠运动作用的机制之一。     

电浮针对原发性痛经镇痛作用的随机对照研究

目的:观察电浮针疗法对原发性痛经(PD)的镇痛作用及临床疗效。方法:171例PD患者,随机分为电浮针组、浮针组和药物组各57例,电浮针组采用电针刺激(疏波频率约60 Hz、串长2.5 s,密波频率约60 Hz、串长5 s,强度2~3 V,持续30 min)与浮针疗法相结合,远取三阴交穴治疗,浮针组采用浮针疗法取三阴交穴治疗,药物组采用口服布洛芬缓释胶囊(0.3 mg/次,2次/日)治疗,3组均连续治疗3个月经周期,治疗后随访3个月经周期统计疗效。结果:纳入171例患者,163例完成试验,脱落8例。电浮针组、浮针组与药物组总有效率(ITT/PP)分别为94.74%/96.43%、91.23%/96.30%和77.19%/83.02%;电浮针组:近期治愈40/40,显效9/9,好转5/5,无效3/2;浮针组:近期治愈38/38,显效10/10,好转4/4,无效5/2;药物组:近期治愈10/10,显效27/27,好转7/7,无效13/9。ITT和PP分析结果一致:电浮针和浮针组的总疗效以及痊愈率显著优于药物组(P<0.01),电浮针与浮针组的总疗效以及痊愈率差异无显著性意义;短时间内电浮针组镇痛作用优于浮针组(P<0.05)。结论:三阴交穴电浮针疗法对PD的镇痛作用具有高效快捷、安全无痛的特点,临床疗效显著优于口服布洛芬缓释胶囊。     

耳穴综合疗法对无先兆偏头痛患者血浆5-HT、SP、β-EP的影响

目的:观察耳穴综合疗法治疗无先兆偏头痛对血浆5-HT、SP和-βEP的影响。方法:对60例无先兆偏头痛患者随机分为两组,治疗组采用耳穴综合疗法,对照组采用尼莫地平等药物口服,在进行临床疗效评价的基础上,对治疗前后5-HT、SP和-βEP进行检测。结果:治疗组总有效率93.33%,对照组总有效率76.67%,两组比较有显著性差异(P<0.05);治疗组5-HT、SP和-βEP含量与治疗前相比有显著性差异(P<0.05),对照组5-HT、SP含量与治疗前相比亦有显著性差异(P<0.05,P<0.01),-βEP前后比较无显著性差异(P>0.05)。结论:耳穴综合疗法治疗无先兆偏头痛主要是通过降低5-HT、SP和升高-βEP达到治疗目的的。     

SP600125抑制c-Jun的激活拮抗肾缺血/再灌注诱导的细胞凋亡

目的应用JNK抑制剂SP600125研究转录因子c-Jun的激活对肾缺血/再灌注（I/R）诱导细胞凋亡的影响。方法采用双侧夹闭大鼠肾蒂缺血45 min后再灌注3 h的方法制备肾I/R动物模型。免疫组化法分析转录因子c-Jun的激活变化情况;原位末端标记法（TUNEL）检测肾小管上皮细胞凋亡情况。结果再灌注3h,p-c-Jun的免疫活性显著增强,凋亡的细胞数明显增加。SP600125明显抑制了p-c-Jun的免疫活性且明显减少肾I/R诱导的细胞凋亡。结论肾I/R诱导的细胞凋亡与c-Jun的激活有关,SP600125能明显抑制转录因子c-Jun的激活,减轻肾小管上皮细胞的凋亡。     

DEEP INSERTION OF NEEDLE AT DAHENG (SP 15) FOR TREATMENT OF 66 CASES OF UROSCHESIS

The author treated 66 cases of uroschesis by adopting deep insertion of needle atDaheng (SP 15) acupoint. Results showed that 64 cases were cured and 2 ineffective, with the totaleffective rate of 96. 15 %.