目的 探讨小乌桂汤治疗类风湿关节炎(RA)的作用机制.方法 60只雄性DBA/1小鼠随机分为正常组8只及造模组52只.造模组采用尾根部及背部分3点皮内注射鸡Ⅱ型胶原蛋白乳化液建立胶原诱导性关节炎(CIA)小鼠模型.造模成功的40只小鼠随机分成模型组、甲氨蝶呤组、小乌桂汤低剂量组、小乌桂汤中剂量组、小乌桂汤高剂量组,每组8只.甲氨蝶呤组予0.1 mg/ml甲氨蝶呤药液2mg/kg灌胃,每周1次;小乌桂汤低、中、高剂量组分别予含生药19.5、39、75g/ (kg·d)小乌桂汤灌胃,每日1次;正常组与模型组给予生理盐水0.2 ml/10 g灌胃,每日1次.4周后评定关节炎指数评分,测量发病足厚度,观察踝关节组织病理并进行组织病理学评分;检测脾脏辅助性T细胞17 (Th17)、调节性T细胞(Treg)及血浆肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素17(IL-17)、白细胞介素1β(IL-1β)、白细胞介素10 (IL-10)的水平.结果 与模型组比较,甲氨蝶呤组及小乌桂汤中、高剂量组关节炎指数评分,发病足厚度,脾脏Th17细胞水平,血浆TNF-α、IL-6、IL-17、IL-1β水平,关节病理总评分,血管翳及骨破坏评分均显著降低,Treg细胞水平均显著升高,甲氨蝶呤组及小乌桂汤高剂量组Th17/Treg值、滑膜炎评分显著降低(P ＜0.05或P＜0.01).与甲氨蝶呤组比较,小乌桂汤低剂量组脾脏Th17细胞水平、Th17/Treg值、关节病理总评分、血管翳及骨破坏评分显著升高,小乌桂汤低、中剂量组Treg细胞水平显著降低(P＜0.01).与小乌桂汤低剂量组比较,小乌桂汤中、高剂量组Th17细胞水平,Th17/Treg值显著降低,Treg细胞水平显著升高,小乌桂汤高剂量组病理总评分、血管翳评分显著降低(P＜0.05或P＜0.01).与小乌桂汤中剂量组比较,小乌桂汤高剂量组Treg细胞水平显著升高(P＜0.01).结论 小乌桂汤对CIA小鼠具有抗炎及免疫调节作用,以高剂量效果最佳,其作用机制与调节Th17/Treg的平衡,抑制TNF-α、IL-6、IL-17、IL-1β炎症因子的表达相关.

OBJECTIVE:To explore the mechanism of Xiaowugui decoction (XWGD) decoction in treating rheumatoid arthritis (RA) in mice.METHODS:Healthy male DBA/1 mice were used for CIA modeling. Twenty-five CIA mice with successful modeling and similar arthritis index (AI) scores were randomized equally into model group (CIA), methotrexate (MTX) group, and low-, medium-, and high-dose XWGD groups (0.975, 1.95, and 3.9 g/mL, respectively), with another 5 normal mice as the normal control group. The mice in normal control and CIA groups were given saline once a day, those in MTX group were given 0.1 mg/mL MTX once a week, and those in XWGD groups were treated daily via garage of XWGD containing crude drugs of different doses for 28 consecutive days. The AI score and HE staining were used to evaluate the changes in the joints of the CIA mice. The effect of XWGD on Th1, Th17, MDSC, G-MDSC and M-MDSC cells were evaluated with flow cytometry.RESULTS:Treatment with MTX and different doses of XWGD significantly decreased the AI score of the mice and relieved joint inflammation as compared with the model group (P < 0.05), and a higher dose of XWGD decoction produced a stronger therapeutic effect. Compared with those in CIA model group, the mice in MTX and XWGD treatment groups showed significantly decreased percentages of Th1, Th17 and M-MDSC cells in the spleen and increased percentages of G-MDSC cells (P < 0.01), and these changes were more conspicuous with a higher dose of XWGD. Correlation analysis showed that Th1 and Th17 cells were positively correlated with M-MDSC and negatively correlated with G-MDSC cells (P < 0.01).CONCLUSIONS:XWGD can improve joint inflammation in CIA mice by increasing the percentages of G-MDSC cells and decreasing the percentages of M-MDSC, Th1 and Th17 cells, and a high dose of XWGD can produce an equivalent therapeutic effect to methotrexate but with better safety.

Excessive degradation of the cartilage articular extracellular matrix (ECM) in chondrocytes has been considered as an important pathological characteristics of OA. In the present study, we demonstrate that the G protein-coupled receptor GPR39 is expressed on SW1353 chondrocytes and is significantly downregulated in response to advanced glycation end products (AGEs). Our findings show that agonism of GPR39 exerts significant protective effects against AGE-induced degradation of articular extracellular matrix. Agonism of GPR39 rescued degradation of type II collagen by decreasing expression of the collagen-degrading enzymes matrix metalloproteinase (MMP)-3 and MMP-13. Additionally, agonism of GPR39 rescued AGE-induced suppression of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2. Agonism of GPR39 prevented degradation of aggrecan by downregulating AGE-induced expression of a disintegrin and metalloproteinase with type I thrombospondin motif (ADAMTS)-4 and ADAMTS-5. Finally, we demonstrate that the effects of GPR39 are mediated through the p38 mitogen activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) cellular signaling pathway. Taken together, our findings show for the first time that targeted therapies involving GPR39 may provide a novel approach for the prevention and treatment of osteoarthritis.

“以学生为中心”及“以病人为中心”的教学导向是目前国内外医学教育教学模式改革的新趋势.中医内科学临床见习是以培养学生的中医临床思维与技能为主要目的课程,本项目遵循临床见习的教学规律,整合中医内科学国家精品资源共享课网络教学平台、临床见习CTPF教学模式及网络即时通讯信息平台等多种教学模式,构建中医内科学临床见习路径,从课前资源整合、课中组织实施、课后持续教育及效果反馈等四部份持续调动学生主观能动性,提高了教学效果.

This study aimed to investigate the expression of miR-146a,miR-155,Ets-1 and IRAK1 in the peripheral blood mononuclear cell(PBMC) of patients with rheumatoid arthritis(RA) and their correlation with the disease activity.PBMC were separated from 44 RA patients and 22 healthy individuals.The levels of miR-146a,miR-155,Ets-1 and IRAK1 were measured using quantitative real-time polymerase chain reaction (qRT-PCR),and their correlation with the clinical and laboratory parameters of RA were analyzed.The relative expression levels of miR-146a,miR-155 were higher in PBMC of RA patients than in normal control group(P＜0.05),and were significantly higher in the active RA patients group than in the low disease activity group(P＜0.05).There was no difference in the levels of miR-146a and miR-155 between the low disease activity group and normal control group(P＞0.05).There was no significant difference in expression level of Ets-1 between the patient group and normal control group(P＜0.05) in a comprehensive (total) comparison.However,the expression level of Ets-1 was lower in high disease activity group than that in the low disease activity group and normal control group(P＜0.05),while there was no difference between the low disease activity group and normal control group (P＞0.05).Compared to the control group,the expression of IRAK1 in PBMC was significantly lower in the high disease activity group and low disease activity group (P＜0.05).However,there was no difference between the two activity groups (P＞0.05).The expression levels of miR-155 was positively correlated to ESR(r=0.319,P=0.042),and negatively correlated to Hb(r=-0.386,P=0.017).The expression levels of Ets-1 were negatively correlated to CRP(r=-0.408,P=0.007).The expression levels of IRAK1 were negatively correlated toRF,DAS28,total complement (r=-0.513,P=0.001;r=-0.332,P =0.029;r=0.490,P=0.015).In conclusion,the expression levels of miR 146a and miR-155 were up-regulated and the expression levels of Ets-1 and IRAK1 were down-regulated in patients with RA with high disease activity.Considering the limitation of sample size in this study,future study with larger sample size is needed to confirm the relationship between miR-146a,miR-155,Ets-1,IRAK1 and RA.Furthermore,studies focusing on the related mechanisms of these molecules in RA are worth to be done.

Objective To observe the clinical efficacy of Lumbago decoction, the experienced prescription of Professor Chen Bao-tian, in treating lumbar disc herniation (LDH) and in order to preliminarily analyze the advantage of traditional Chinese medicine in treating LDH.Methods A total of 60 patients with LDH who met the inclusion criteria were randomly divided into treatment group (n=30) and control group (n=30). The Lumbago decoction was orally given in the treatment group, while Celebrex was given in the control group; the course of treatment was 28 days. The two groups were compared in term of overall response rate and main symptom scores.Results Within these two groups, there was a significant difference in response rate (P<0.05), the response of treatment group was better than control group. After treated for 14 days, the total symptoms score of treatment group was better than control group (P<0.05). After treated for 28 days, the total symptoms score, release of low back pain and release of leg pain in treatment group was better than in control group (P<0.05). Conclusion Lumbago decoction has a better clinical effect compared with Celebrex in the treatment of LDH and is superior to meloxicam in benefit symptom scores. This prescription shows promise for clinical application.

OBJECTIVES:Although myeloid-derived suppressor cells (MDSCs) have been linked to T cell tolerance, their role in autoimmune rheumatoid arthritis (RA) remains elusive. Here we investigate the potential association of MDSCs with the disease pathogenesis using a preclinical model of RA and specimen collected from patients with RA.METHODS:The frequency of MDSCs in blood, lymphoid tissues, inflamed paws or synovial fluid and their association with disease severity, tissue inflammation and the levels of pathogenic T helper (Th) 17 cells were examined in arthritic mice or in patients with RA (n=35) and osteoarthritis (n=15). The MDSCs in arthritic mice were also characterised for their phenotype, inflammation status, T cell suppressive activity and their capacity of pro-Th17 cell differentiation. The involvement of MDSCs in the disease pathology and a Th17 response was examined by adoptive transfer or antibody depletion of MDSCs in arthritic mice or by coculturing mouse or human MDSCs with naïve CD4+ T cells under Th17-polarising conditions.RESULTS:MDSCs significantly expanded in arthritic mice and in patients with RA, which correlated positively with disease severity and an inflammatory Th17 response. While displaying T cell suppressive activity, MDSCs from arthritic mice produced high levels of inflammatory cytokines (eg, interleukin (IL)-1β, TNF-α). Mouse and human MDSCs promoted Th17 cell polarisation ex vivo. Transfer of MDSCs facilitated disease progression, whereas their elimination in arthritic mice ameliorated disease symptoms concomitant with reduction of IL-17A/Th17 cells.CONCLUSIONS:Our studies suggest that proinflammatory MDSCs with their capacity to drive Th17 cell differentiation may be a critical pathogenic factor in autoimmune arthritis.

Both selenium (Se) and polysaccharides from Pyracantha fortuneana (Maxim.) Li (PFPs) (P. fortuneana) have been reported to possess antioxidative and immuno-protective activities. Whether or not Se-containing polysaccharides (Se-PFPs) have synergistic effect of Se and polysaccharides on enhancing the antioxidant and immune activities remains to be determined. We previously reported that polysaccharides isolated from Se-enriched P. fortuneana (Se-PFPs) possessed hepatoprotective effects. However, it is not clear whether or not they have anti-mutagenic effects. In the present study, we compared and evaluated anti-mutagenic effects of Se-PFPs at three concentrations (1.35, 2.7 and 5.4 g/kg body weight) with those of PFPs, Se alone or Se + PFPs in mice using micronucleus assay in bone marrow and peripheral blood as well as mitomycin C-induced chromosomal aberrations in mouse testicular cells. We also elucidated the underlying mechanism. Our results demonstrated that Se-PFPs inhibited cyclophosphamide (CP)-induced micronucleus formation in both bone marrow and peripheral blood, enhanced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in mouse liver, and reduced the activity and expression of cytochrome P450 1A (CYP4501A) in mouse liver in a dose-dependent manner. In addition, we found that the anti-mutagenic potential of Se-PFPs was higher than those of PFPs, Se alone or Se + PFPs at the same level. These results suggest that the anti-mutagenic potential of Se-PFPs may be mediated through the inhibition of the activity and expression of CYP4501A. This study indicates that application of Se-PFPs may provide an alternative strategy for cancer therapy by targeting CYP1A family.

INTRODUCTION:Infection of pathogenic microorganisms is an important reason for autoimmune diseases (ADs). Interleukin-1 (IL-1) receptor-associated kinase-1 (IRAK1) is a key mediator in infection immunity, while the gene of IRAK1 is recognized as a risk gene in ADs. Three single nucleotide polymorphisms (SNPs) in IRAK1 (rs3027898, rs1059702, rs1059703) are considered to be associated with ADs risk. However, the results are conflicting. We conducted this study to get more precise estimations.METHODOLOGY:PubMed, OvidSP, and CNKI databases (published prior to August 2014) were searched, and data was extracted from eligible studies. The procedure of statistical analysis was performed using STATA 12.0 software. A random effect model or fixed effect model was chosen based on the between-study heterogeneities.RESULTS:Of the studies involved, 11 studies included 10,705 cases (9,865 controls) for rs3027898, 9 studies included 15,005 cases (14,997 controls) for rs1059702, and 7 studies included 8,115 cases (6,815 controls) for rs1059703. Overall, the results showed that there were significant associations with ADs risk in three genetic models for rs3027898 and in four genetic models for rs1059702, but in neither model for rs1059703. Moreover, in stratified analyses, different extents of associations were found in some different genetic models for all three SNPs.CONCLUSION:Our data demonstrated that these three SNPs (rs3027898, rs1059702, rs1059703) in IRAK1 were associated with ADs risk.

V-ets erythroblastosis virus E26 oncogene homolog 1 (ETS1) is recognized as a gene of risk to autoimmune diseases (ADs). Two single nucleotide polymorphisms (SNPs) in ETS1 (rs1128334 G>A and rs10893872 T>C) were considered associated with ADs risk. However, the results remain conflicting.We performed a meta-analysis to evaluate more precise estimations of any relationship. We searched PubMed, OvidSP, and Chinese National Knowledge Infrastructure databases (papers published prior to September 12, 2014) and extracted data from eligible studies. Meta-analysis was performed using the STATA 12.0 software. Random effect model or fixed effect model were chosen according to the study heterogeneities.A total of 11 studies including 7359 cases (9660 controls) for rs1128334 and 8 studies including 5419 cases (7122 controls) for rs10893872 were involved in this meta-analysis. Overall, our results showed that there were significant associations for rs1128334 with AD risk in 5 genetic models, both in pooled analysis and in systemic lupus erythematous (SLE) subgroup, and in 3 genetic models of the uveitis subgroup. Although for rs10893872, the results showed that there were significant associations in allele model both in pooled analysis and in SLE subgroup. As a conclusion, this meta-analysis demonstrated that these 2 SNPs (rs1128334 and rs10893872) in ETS1 were associated with ADs risk.

BackgroundPublished data showed that the susceptibility of autoimmune diseases (ADs) was associated with the polymorphism rs2910164 in microRNA-146a (miR-146a). However, the results remain controversial so far. Two meta-analyses published in 2013 and 2014 came to opposite conclusions. In order to derive a more precise estimation of the relationship, we performed this meta-analysis.MethodsWe searched the PubMed, OvidSP and CNKI databases (published prior to September 8th, 2014) and extracted data from eligible studies. The procedure of meta-analysis was performed by using the Stata 12.0 software. Random effect model or fixed effect model were chosen respectively, according to the between study heterogeneities.ResultsA total of 24 case-control studies, 11 more than previous meta-analysis on this topic, were involved. We took stratified analyses by different ethnicities and different types of diseases in different genetic models. In Caucasian subgroup, significant increased risks of GC genotype and GC+CC genotype with ADs susceptibility were found in heterozygote model (GC vs GG, OR = 1.38, 95% CI 1.04-1.83, p = 0.024) and dominant model (GC+CC vs GG, OR = 1.37, 95% CI 1.01-1.85, p = 0.041), respectively. Meanwhile, in other disease subgroup, significant increased risks of C allele, CC genotype and GC+CC genotype were found in allele model (C vs G, OR = 1.16, 95% CI 1.04-1.31, p = 0.010), homozygote model (CC vs GG, OR = 1.42, 95% CI 1.10-1.84, p = 0.006) and dominant model (GC+CC vs GG, OR = 1.25, 95% CI 1.04-1.51, p = 0.020), respectively.ConclusionsMiR-146a rs2910164 G>C polymorphism was associated with the susceptibility of ADs.

Saikosaponin a (SSa), a main constituent of the Chinese herb Bupleurum chinense DC., has been demonstrated to have antiepileptic activity. Recent studies have shown that SSa could inhibit NMDA receptor current and persistent sodium current. However, the effects of SSa on potassium (K+) currents remain unclear. In this study, we tested the effect of SSa on 4AP-induced epileptiform discharges and K+ currents in CA1 neurons of rat hippocampal slices. We found that SSa significantly inhibited epileptiform discharges frequency and duration in hippocampal CA1 neurons in the 4AP seizure model in a dose-dependent manner with an IC50 of 0.7 μM. SSa effectively increased the amplitude of ITotal and IA, significantly negative-shifted the activation curve, and positive-shifted steady-state curve of IA. However, SSa induced no significant changes in the amplitude and activation curve of IK. In addition, SSa significantly increased the amplitude of 4AP-sensitive K+ current, while there was no significant change in the amplitude of TEA-sensitive K+ current. Together, our data indicate that SSa inhibits epileptiform discharges induced by 4AP in a dose-dependent manner and that SSa exerts selectively enhancing effects on IA. These increases in IA may contribute to the anticonvulsant mechanisms of SSa.

目的 用生物信息学方法探讨TNF-α抑制剂治疗类风湿关节炎的相关基因，为探讨TNF-α抑制剂治疗类风湿关节炎的机制提供新思路。 方法 从基因芯片公共数据库Gene Expression Omnibus (GEO)中下载TNF-α抑制剂治疗类风湿关节炎的相关基因芯片数据，利用String、KEGG、Panther等在线分析软件对差异表达基因进行生物信息学分析。 结果 在60个差异表达基因所编码的蛋白中，有29个蛋白与其他蛋白存在相互作用关系，作用过程涉及多种生物学通路，与多个生物学过程和分子功能相关。 结论 TNF-α抑制剂治疗类风湿关节炎涉及多基因相互作用，与NF-κB信号通路、MAPK信号通路关系密切。

<正>研究生是我国优秀青年人才的来源,研究生的培养质量关系到我国的人才储备。对于临床医学相关专业研究生,他们可能即将成为从事医疗服务的临床医师,也可能成为从事医学相关研究的科研人员,还可能成为临床医学院校教书育人的老师;因此,临床医学相关专业的研究生培养目标中常包含"具有独立承担本学科科研、教学和临床医疗工作的能力"等方面的要求。基于目前医学高等院

OBJECTIVE To construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells. METHODS The total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies. RESULTS The libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11). CONCLUSION We have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR) to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS) analysis, as well as in conditioned medium in a secreted version for function analysis.

OBJECTIVETo construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).METHODSThe total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.RESULTSUsing 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.CONCLUSIONThe personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.

本文对青年教师如何提升中医内科学教学能力进行了分析思考及总结.青年教师需要扎实提高临床基本功、教学基本功,努力学习多种教学技巧;在中医内科学教学过程中,掌握课程的重点,并合理讲解;紧跟教学发展方向,合理运用各种多媒体教学手段,从而最终提升自身的中医内科学教学能力.

Objective To investigate the genes associated with Alzheimer's disease (AD) by bioinformatics methods to provide new thoughts for basic research and clinical treatment of AD.Methods The microarray data of AD-related genes were downloaded from the Gene Expression Omnibus (GEO) database and analyzed by bioinformatics methods using String,KEGG and Panther databases.Results Of all the 34 differentially expressed genes,25 were found to encode proteins with interactions; they referred to multiple biological pathways,biological processes and molecular functions.Conclusion The pathogenesis of AD involves multiple genes,especially genes with Ca2+ signaling,and investigations of these genes may provide valuablc insights into the mechanism of AD.

《中药学》教学中涉及的药材种类繁多,教学过程中学生易于感到枯燥无味,学习热情不高。与传统教学相比,应用计算机互动教室进行教学,有利于师生之间的及时互动交流和反馈,有利于激发学生参与热情,培养学生的中医药科学素养。同时,利用先进的技术手段帮助学生建立教学成果档案袋,能有效促进教学效果的提升。