Testing the role of genetic variation of the MC4R gene in Chinese population in antipsychotic-induced metabolic disturbance

Antipsychotic-induced metabolic disturbance(AIMD) is a common adverse effect of antipsychotics with genetics partly underpinning variation in susceptibility among schizophrenia patients. Melanocortin4 receptor(MC4 R) gene, one of the candidate genes for AIMD, has been under-studied in the Chinese patients. We conducted a pharmacogenetic study in a large cohort of Chinese patients with schizophrenia. In this study, we investigated the genetic variation of MC4 R in Chinese population by genotyping two SNPs(rs489693 and rs17782313) in 1,991 Chinese patients and examined association of these variants with the metabolic effects that were often observed to be related to AIMD. Metabolic measures, including body mass index(BMI), waist circumference(WC), glucose, triglyceride, high-density lipoprotein(HDL), and low-density lipoprotein(LDL) levels were assessed at baseline and after 6-week antipsychotic treatment. We found that interaction of SNP×medication status(drug-na?ve/medicated) was significantly associated with BMI, WC, and HDL change %, respectively. Both SNPs were significantly associated with baseline BMI and WC in the medicated group. Moderate association of rs489693 with WC, Triglyceride, and HDL change % were observed in the whole sample. In the drug-na?ve group, we found recessive effects of rs489693 on BMI gain more than 7%, WC and Triglyceride change %, with AA incurring more metabolic adverse effects. In conclusion, the association between rs489693 and the metabolic measures is ubiquitous but moderate. Rs17782313 is less involved in AIMD. Two SNPs confer risk of AIMD to patients treated with different antipsychotics in a similar way.     

番茄Sly-MIR167的抗冷性研究

以番茄为材料,采用Northern杂交技术,分析番茄MIR167(Sly-MIR167)在低温胁迫下的表达模式,以明确Sly-MIR167在冷胁迫下的分子调控机制,为基因工程在改良番茄品种中的实际应用提供依据。结果显示:(1)25℃下Sly-MIR167在番茄根、茎、花瓣、果实、叶片都有表达,4℃低温胁迫下的表达量均增加,表明Sly-MIR167表达受低温诱导。(2)采用农杆菌侵染构建表达载体并转化番茄获得转基因植株,冷胁迫实验结果显示:转基因植株在冷胁迫处理的生长状况明显优于对照;另外,冷胁迫下2个转基因株系(T2-5和T2-19)的最大光化学效率、叶绿素含量下降幅度明显低于野生型;脯氨酸含量高于野生型;MDA含量低于野生型,表明Sly-MIR167能够提高番茄对冷胁迫的耐受性。(3)通过miRU在线软件预测Sly-MIR167的靶基因为NF-YA1、NF-YA2,利用RT-PCR技术分析其表达下调,证明它们被MIR167负调控。     

Tracking miRNA precursor metabolic products and processing sites through completely analyzing high-throughput sequencing data

The small non-coding important regulatory molecules, microRNAs (miRNAs), have been widely and deeply studied especially combining high-throughput sequencing technologies. Here, we attempted to track detailed miRNA precursor metabolic products and gain further insight into pre-miRNA processing by completely analyzing high-throughput sequencing data. Highly expressed miRNA precursors could be entirely covered by various short RNAs and small RNA fragments with a hierarchical distribution. miRNAs and some miRNA* regions were detected quite abundant short RNAs as expected, while other regions of precursors were found shorter RNAs or small fragments with fewer sequence counts. Furthermore, we developed a method to analyze relative expression levels of special RNA classes according to divergence of 5′ and 3′ ends, respectively. Generally, there were several quite abundant RNA classes from a given miRNA locus, which suggested dominant cleavage sites of Drosha and Dicer during pre-miRNA processing. Compared with 3′ end, dominant cleavage site in 5′ end always focused on a specific position, which ensured conservation of the identity of miRNA (5′-seed sequence, nucleotides 2–8). Overall, a comprehensive analysis of sequencing data can be used to track pre-miRNA metabolic products and mechanism of pre-miRNA processing and metabolism.     

miR-134 inhibits epithelial to mesenchymal transition by targeting FOXM1 in non-small cell lung cancer cells

Recent studies have implied that miRNAs act as crucial modulators for epithelial-to-mesenchymal transition (EMT). We found that miR-134 expression correlated with invasive potential and EMT phenotype of NSCLC cells. Functional assays demonstrated that miR-134 inhibited EMT in NSCLC cells. In addition, we showed that Forkhead Box M1 (FOXM1) is a direct target of miR-134. Knockdown of FOXM1 reversed EMT resembling that of miR-134 overexpression. We further found that FOXM1 was involved in TGF-β1-induced EMT in A549 cells. These findings suggest that miR-134 acts as a novel EMT suppressor in NSCLC cells.     

Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells

Accumulating evidence has shown that dysfunctional mitochondria can be selectively removed by mitophagy. Dysregulation of mitophagy is implicated in the development of neurodegenerative disease and metabolic disorders. How individual mitochondria are recognized for removal and how this process is regulated remain poorly understood. Here we report that FUNDC1, an integral mitochondrial outer-membrane protein, is a receptor for hypoxia-induced mitophagy. FUNDC1 interacted with LC3 through its typical LC3-binding motif Y(18)xxL(21), and mutation of the LC3-interaction region impaired its interaction with LC3 and the subsequent induction of mitophagy. Knockdown of endogenous FUNDC1 significantly prevented hypoxia-induced mitophagy, which could be reversed by the expression of wild-type FUNDC1, but not LC3-interaction-deficient FUNDC1 mutants. Mechanistic studies further revealed that hypoxia induced dephosphorylation of FUNDC1 and enhanced its interaction with LC3 for selective mitophagy. Our findings thus offer insights into mitochondrial quality control in mammalian cells.     

A regulatory effect of IL-21 on T follicular helper-like cell and B cell in rheumatoid arthritis

Introduction

Interleukin (IL)-21 is a member of type I cytokine family. Recent studies indicate that IL-21 can promote T follicular helper (Tfh) cell differentiation and survival, a specialized T cell subset which provides help for B cell. It can also regulate the activation, proliferation and differentiation of human B cell and immunoglobulin (Ig) production as well as isotype switching of plasma cell. Rheumatoid arthritis (RA) is characterized by auto-antibodies overproduction such as rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) antibody, suggesting a pivotal role of Tfh cell and B cell in the pathogenesis of RA. This study aimed to investigate whether IL-21 had a regulatory effect on Tfh cell and B cell in RA.

Methods

Serum IL-21 concentrations were measured by ELISA. The correlations between serum IL-21 levels and clinical features of RA patients were analyzed by Spearman''s rank test. The percentages of Tfh-like cells, IL-21 receptor (R) expression on Tfh-like cells and B cells in peripheral blood (PB) were analyzed by flow cytometry. Peripheral blood mononuclear cells (PBMC) were stimulated by rIL-21 (100 ng/ml) in the presence or absence of anti-CD40 and/or anti-IgM, and changes of IL-21R, activation-associated surface markers (CD25, CD69 and CD40), the proliferation, apoptosis and differentiation of B cells were analyzed by flow cytometry. Production of IgG and IgM in the culture supernatants was determined by ELISA.

Results

The results showed that the serum IL-21 levels in RA patients were significantly higher than that of healthy controls (HC). IL-21 concentrations were positively correlated with 28-joint count disease activity score (DAS28) and anti-CCP antibody in RA patients with high IL-21 levels. Furthermore, the frequencies of peripheral CXCR5⁺PD-1⁺CD4⁺ Tfh-like cells markedly increased in RA patients and the percentages of Tfh-like cells were positively correlated with DAS28 and anti-CCP antibody levels. Moreover, elevated IL-21 levels were also correlated with the frequencies of Tfh-like cells. IL-21R expression on both Tfh-like cells and B cells were significantly enhanced in RA patients. In cultures vitro, exogenous IL-21 upregulated IL-21R expression and activation-associated surface markers on B cells and promoted more B cell proliferation in RA than in HC. This IL-21-mediated effect could be reversed by IL-21R-specific neutralizing antibody. Importantly, IL-21 promoted more differentiation of B cell into plasmablast and higher levels of IgG and IgM production in RA than in HC.

Conclusions

Increased serum IL-21 levels in RA patients correlate with DAS28, anti-CCP antibody and frequencies of Tfh-like cells. IL-21 supports B cell activation, proliferation and antibody secretion via IL-21R pathway. Thus, IL-21 may be involved in the pathogenesis of RA and antagonizing IL-21 could be a novel strategy for the therapy of RA.     

中国西南干旱河谷植物群落的数量分类和排序分析

中国西南干旱河谷植被是我国西南横断山区特有的植被类型, 目前关于西南干旱河谷植被还没有整体性的群落类型划分研究。根据对甘肃、四川、云南三省九条主要河流的干旱河谷段野外调查和文献来源的1,339个植物群落样方数据, 采用自适应仿射传播聚类方法, 对我国西南干旱河谷的植物群落进行数量分类, 并采用典范对应分析方法进行排序分析。结果表明: (1)调查样方的植物群落分为7个植被型(稀树草原、肉质灌丛、常绿阔叶灌丛、暖性落叶阔叶灌丛、常绿硬叶林、落叶阔叶林和暖性针叶林), 24个群系, 31个群丛类型。暖性落叶阔叶灌丛是本植被区的代表性植被类型; 分布最广的群系为鞍叶羊蹄甲灌丛(Form. Bauhinia brachycarpa, 样方比例50.9%)、黄茅灌草丛(Form. Heteropogon contortus, 样方比例11.9%)、孔颖草灌草丛(Form. Bothriochloa pertusa, 样方比例5.6%)、黄荆灌丛(Form. Vitex negundo, 样方比例4.2%)、知风草灌草丛(Form. Eragrostis ferruginea, 样方比例3.8%)、车桑子灌丛(Form. Dodonaea viscosa, 样方比例3.4%)、云南松疏林(Form. Pinus yunnanensis, 样方比例3.3%)。(2)冬季低温和降水的季节性是限制干旱河谷植物群落分布的主要气候因子。稀树草原、肉质灌丛是典型的干热河谷植被类型; 暖性落叶阔叶灌丛、常绿硬叶林、常绿阔叶灌丛是干暖河谷植被的优势类型; 暖性针叶林、落叶阔叶林则主要在干温河谷环境占优势。     

三江并流地区干旱河谷植物物种多样性海拔梯度格局比较

在滇西北三江并流地区典型干旱河谷段, 在怒江、澜沧江和金沙江的东、西坡共设置了6条海拔梯度样带, 通过标准样地的植物群落调查, 分析各条样带植物的物种丰富度、物种更替率的海拔梯度格局, 并比较了地理和植被变量对分布格局的解释。干旱河谷植被带位于海拔3,000 m以下, 以灌丛和灌草丛为主, 其在各河谷的分布上限自西向东依次升高。植物物种丰富度的分布主要与海拔、流域、经纬度和植被带有关, 沿纬度和海拔梯度升高而显著增加的格局主要表现在草本层和灌木层, 灌木物种丰富度还呈现自西向东显著增加的趋势。怒江的灌木和草本种物种丰富度显著高于金沙江和澜沧江, 三条江的乔木种丰富度差异则不显著。森林带的样方草本物种丰富度显著低于灌草丛带样方, 并且还拥有后者没有的乔木种。不同样带的植物物种更替速率呈现了不一致的海拔梯度格局, 但均在样带海拔下部的灌草丛群落与海拔上部森林群落之间的交错带出现峰值。森林-灌草丛植被交错带在怒江样带处于海拔1,900-2,100 m处, 在澜沧江河谷位于海拔2,300-2,400 m, 在金沙江河谷位于海拔2,700-2,900 m。所有海拔样带的森林段或灌草丛段相对于同一样带不同植被段之间的物种更替程度为最小, 不仅小于同一流域不同样带相同植被段之间物种更替率的均值, 更小于所有样带相同植被段之间的更替率均值。在三条河流6条海拔样带的12个植被带段之间的物种更替变化中, 空间隔离因素可以解释34.2%, 而植被类型差异仅能解释不到0.5%。本研究结果显示了环境差异对不同植被类型物种丰富度的首要影响, 和各河流之间的空间隔离对植物群落构建和物种构成的主要作用。     

Apoptotic cell-mediated suppression of streptococcal cell wall-induced arthritis is associated with alteration of macrophage function and local regulatory T-cell increase: a potential cell-based therapy?

Introduction  

Experimental streptococcal cell wall (SCW)-induced arthritis is characterized by two successive phases of the disease. The acute phase occurs early and is associated with an inflammatory process and neutrophil infiltration into the synovium. The second chronic phase is related to effector T-cell activation and the dysregulation of macrophage function. Creation of an immunomodulatory environment has been attributed to apoptotic cells themselves, apoptotic cell uptake by phagocytes as well as a less sensibility of phagocytes capturing apoptotic bodies to activation. Therefore we evaluated the potential of apoptotic cell injection to influence the course of inflammation in SCW-induced arthritis in rats.     

CD3-specific antibody-induced immune tolerance involves transforming growth factor-beta from phagocytes digesting apoptotic T cells

Intact CD3-specific antibody transiently depletes large numbers of T cells and subsequently induces long-term immune tolerance. The underlying mechanisms for the systemic tolerance, however, remain unclear. We show here that treatment of normal mice with intact antibody to CD3 increases systemic transforming growth factor-beta (TGF-beta) produced by phagocytes exposed to apoptotic T cells. Among the phagocytes, macrophages and immature dendritic cells (iDCs) secrete TGF-beta upon ingestion of apoptotic T cells, which induces CD4+Foxp3+ regulatory T cells in culture and contributes to immune tolerance mediated by CD3-specific antibody in vivo. In accordance with these results, depletion of macrophages and iDCs not only abrogates CD3-specific antibody-mediated prevention of myelin oligodendrocyte glycoprotein-induced acute experimental autoimmune encephalomyelitis (EAE), but also reverses the therapeutic effects of antibody to CD3 on established disease in a model of relapsing-remitting EAE. Thus, CD3-specific antibody-induced immune tolerance is associated with TGF-beta production in phagocytes involved in clearing apoptotic T cells, which suggests that apoptosis is linked to active suppression in immune tolerance.