T‐bet is essential for Th1‐mediated,but not Th17‐mediated,CNS autoimmune disease

T cells that produce both IL‐17 and IFN‐γ, and co‐express ROR‐γt and T‐bet, are often found at sites of autoimmune inflammation. However, it is unknown whether this co‐expression of T‐bet with ROR‐γt is a prerequisite for immunopathology. We show here that T‐bet is not required for the development of Th17‐driven experimental autoimmune encephalomyelitis (EAE). The disease was not impaired in T‐bet^?/? mice and was associated with low IFN‐γ production and elevated IL‐17 production among central nervous system (CNS) infiltrating CD4⁺ T cells. T‐bet^?/? Th17 cells generated in the presence of IL‐6/TGF‐β/IL‐1 and IL‐23 produced GM‐CSF and high levels of IL‐17 and induced disease upon transfer to naïve mice. Unlike their WT counterparts, these T‐bet^?/– Th17 cells did not exhibit an IL‐17→IFN‐γ switch upon reencounter with antigen in the CNS, indicating that this functional change is not critical to disease development. In contrast, T‐bet was absolutely required for the pathogenicity of myelin‐responsive Th1 cells. T‐bet‐deficient Th1 cells failed to accumulate in the CNS upon transfer, despite being able to produce GM‐CSF. Therefore, T‐bet is essential for establishing Th1‐mediated inflammation but is not required to drive IL‐23‐induced GM‐CSF production, or Th17‐mediated autoimmune inflammation.     

小鼠T-bet基因真核表达载体的构建

目的构建小鼠T—bet基因重组真核表达载体pcDNA3一T—bet,为T—bet基因治疗研究提供有效的生物表达系统。方法采用内切酶切从质粒T—bet/GFP—RV中获得约1.7 kb的小鼠T—betcDNA片段,以单酶切非定向克隆方式与真核表达质粒pcDNA3连接,构建真核表达载体pcDNA3-T-bet。结果经PCR筛选阳性重组子,双酶切鉴定T—bet基因重组方向,并测序鉴定无错配及插入移位等DNA顺序改变。结论本实验成功构建了小鼠T—bet基因真核表达载体pcDNA3-T-bet。     

加减龙胆泻肝汤治疗带下病湿热下注的临床研究

目的:探讨加减龙胆泻肝汤治疗带下病湿热下注型患者的临床疗效及其应用价值。方法:选取于2010年7月1日～2013年1月1日在我院收治的带下病湿热下注型患者80例。将患者随机分配为中药治疗组与西药对照组,每组40例患者。同时保证患者具有大致相同的既往病史。中药治疗组采用加减龙胆泻肝汤的治疗方式;而西药对照组,则根据患者阴道感染的种类选择正确的药物治疗。其中对照组中40例患者均为细菌性阴道感染,则采用甲硝唑治疗。对比两种治疗方法的临床效果,并讨论其应用价值。结果:中药治疗组显效13例,有效25例,无效2例,有效率为95.00%;西药对照组显效20例,有效15例,无效5例,有效率为87.50%。同时两组患者带下病症状均得到良好改善,差异无统计学意义(P>0.05)。结论:运用加减龙胆泻肝汤治疗带下病湿热下注型患者,与西药治疗疗效相当,临床效果显著,值得广泛应用及推广。     

Betaine feeding prevents the blood alcohol cycle in rats fed alcohol continuously for 1 month using the rat intragastric tube feeding model

Background

Blood alcohol levels (BAL) cycle up and down over a 7–8 day period when ethanol is fed continuously for one month in the intragastric tube feeding rat model (ITFRM) of alcoholic liver disease. The cycling phenomenon is due to an alternating increase and decrease in the metabolic rate. Recently, we found that S-adenosyl-methionine (SAMe) fed with alcohol prevented the BAL cycle.

Method

Using the ITFRM we fed rats betaine (2 g/kg/day) with ethanol for 1 month and recorded the daily 24 h urine ethanol level (UAL) to measure the BAL cycle. UAL is equivalent to BAL because of the constant ethanol infusion. Liver histology, steatosis and BAL were measured terminally after 1 month of treatment. Microarray analysis was done on the mRNA extracted from the liver to determine the effects of betaine and alcohol on changes in gene expression.

Results

Betaine fed with ethanol completely prevented the BAL cycle similar to SAMe. Betaine also significantly reduced the BAL compared to ethanol fed rats without betaine. This was also observed when SAMe was fed with ethanol. The mechanism involved in both cases is that SAMe is required for the conversion of epinephrine from norepinephrine by phenylethanolamine methyltransferase (PNMT). Epinephrine is 5 to 10 fold more potent than norepinephrine in increasing the metabolic rate. The increase in the metabolic rate generates NAD, permitting ADH to increase the oxidation of alcohol. NAD is the rate limiting factor in oxidation of alcohol by alcohol dehydrogenase (ADH). This explains how SAMe and betaine prevented the cycle. Microarray analysis showed that betaine feeding prevented the up regulation of a large number of genes including TLR2/4, Il-1b, Jax3, Sirt3, Fas, Ifngr1, Tgfgr2, Tnfrsf21, Lbp and Stat 3 which could explain how betaine prevented fatty liver.

Conclusion

Betaine feeding lowers the BAL and prevents the BAL cycle by increasing the metabolic rate. This increases the rate of ethanol elimination by generating NAD.