Exosomes derived from umbilical cord mesenchymal stem cells alleviate viral myocarditis through activating AMPK/mTOR‐mediated autophagy flux pathway

Human umbilical cord mesenchymal stem cell‐derived exosomes (hucMSC‐exosomes) have been implicated as a novel therapeutic approach for tissue injury repair and regeneration, but the effects of hucMSC‐exosomes on coxsackievirus B3 (CVB3)‐induced myocarditis remain unknown. The object of the present study is to investigate whether hucMSC‐exosomes have therapeutic effects on CVB3‐induced myocarditis (VMC). HucMSC‐exosomes were identified using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and Western blot. The purified hucMSC‐exosomes tagged with PKH26 were tail intravenously injected into VMC model mice in vivo and used to administrate CVB3‐infected human cardiomyocytes (HCMs) in vitro, respectively. The effects of hucMSC‐exosomes on myocardial pathology injury, proinflammatory cytokines and cardiac function were evaluated through haematoxylin and eosin (H&E) staining, quantitative polymerase chain reaction (qPCR) and Doppler echocardiography. The anti‐apoptosis role and potential mechanism of hucMSC‐exosomes were explored using TUNEL staining, flow cytometry, immunohistochemistry, Ad‐mRFP‐GFP‐LC3 transduction and Western blot. In vivo results showed that hucMSC‐exosomes (50 μg iv) significantly alleviated myocardium injury, shrank the production of proinflammatory cytokines and improved cardiac function. Moreover, in vitro data showed that hucMSC‐exosomes (50 μg/mL) inhibited the apoptosis of CVB3‐infected HCM through increasing pAMPK/AMPK ratio and up‐regulating autophagy proteins LC3II/I, BECLIN‐1 and anti‐apoptosis protein BCL‐2 as well as decreasing pmTOR/mTOR ratio, promoting the degradation of autophagy flux protein P62 and down‐regulating apoptosis protein BAX. In conclusion, hucMSC‐exosomes could alleviate CVB3‐induced myocarditis via activating AMPK/mTOR‐mediated autophagy flux pathway to attenuate cardiomyocyte apoptosis, which will be benefit for MSC‐exosome therapy of myocarditis in the future.     

参与调控桂花花朵开放的SPL基因鉴定及表达分析

SPL(SQUAMOSA promoter binding protein like)是植物特有的基因家族,在花发育过程中具有重要调控作用。该研究以桂花全基因组数据为基础,对SPL基因家族成员的蛋白理化性质、系统进化、基因结构和不同组织的表达模式进行生物信息学分析,筛选出花组织中较高表达的基因进行实时荧光定量、亚细胞定位及酵母自激活验证,为解析SPL基因参与调控桂花花朵开放过程中的作用机制和功能提供基因资源。结果显示：(1)共鉴定出29个桂花SPL基因家族成员,它们均具有SBP结构域,且不均匀分布在15条染色体上。(2)与拟南芥构建系统进化树,聚类可分为8大亚群,同亚群的OfSPL基因具有高度相似的基因结构。(3)RNA seq数据分析表明,OfSPL9/10/17/19/21/27/28整体FPKM值较高,在花组织中具有一定的特异性;qRT PCR分析表明,OfSPL10/21在花朵开放过程中的相对表达量呈先升后降的趋势。(4)亚细胞定位和转录自激活活性分析显示,OfSPL10/21编码蛋白主要定位于细胞核上,不具有转录自激活活性。研究推测,OfSPL10/21可能参与调控桂花花色与花香物质的生物合成。     

Genome-Wide Analysis of NAC Transcription Factors and Characterization of the Cold Stress Response in Sweet Osmanthus

Plant Molecular Biology Reporter - Sweet osmanthus (Osmanthus fragrans) is an evergreen aromatic woody tree widely used in landscaping. However, O. fragrans is very sensitive to cold stress, which...     

Disruption of Ttll5/Stamp Gene (Tubulin Tyrosine Ligase-like Protein 5/SRC-1 and TIF2-associated Modulatory Protein Gene) in Male Mice Causes Sperm Malformation and Infertility

TTLL5/STAMP (tubulin tyrosine ligase-like family member 5) has multiple activities in cells. TTLL5 is one of 13 TTLLs, has polyglutamylation activity, augments the activity of p160 coactivators (SRC-1 and TIF2) in glucocorticoid receptor-regulated gene induction and repression, and displays steroid-independent growth activity with several cell types. To examine TTLL5/STAMP functions in whole animals, mice were prepared with an internal deletion that eliminated several activities of the Stamp gene. This mutation causes both reduced levels of STAMP mRNA and C-terminal truncation of STAMP protein. Homozygous targeted mutant (Stamp^tm/tm) mice appear normal except for marked decreases in male fertility associated with defects in progressive sperm motility. Abnormal axonemal structures with loss of tubulin doublets occur in most Stamp^tm/tm sperm tails in conjunction with substantial reduction in α-tubulin polyglutamylation, which closely correlates with the reduction in mutant STAMP mRNA. The axonemes in other structures appear unaffected. There is no obvious change in the organs for sperm development of WT versus Stamp^tm/tm males despite the levels of WT STAMP mRNA in testes being 20-fold higher than in any other organ examined. This defect in male fertility is unrelated to other Ttll genes or 24 genes previously identified as important for sperm function. Thus, STAMP appears to participate in a unique, tissue-selective TTLL-mediated pathway for α-tubulin polyglutamylation that is required for sperm maturation and motility and may be relevant for male fertility.     

Genome-Wide Analyses of Radioresistance-Associated miRNA Expression Profile in Nasopharyngeal Carcinoma Using Next Generation Deep Sequencing

Background

Rapidly growing evidence suggests that microRNAs (miRNAs) are involved in a wide range of cancer malignant behaviours including radioresistance. Therefore, the present study was designed to investigate miRNA expression patterns associated with radioresistance in NPC.

Methods

The differential expression profiles of miRNAs and mRNAs associated with NPC radioresistance were constructed. The predicted target mRNAs of miRNAs and their enriched signaling pathways were analyzed via biological informatical algorithms. Finally, partial miRNAs and pathways-correlated target mRNAs were validated in two NPC radioreisitant cell models.

Results

50 known and 9 novel miRNAs with significant difference were identified, and their target mRNAs were narrowed down to 53 nasopharyngeal-/NPC-specific mRNAs. Subsequent KEGG analyses demonstrated that the 53 mRNAs were enriched in 37 signaling pathways. Further qRT-PCR assays confirmed 3 down-regulated miRNAs (miR-324-3p, miR-93-3p and miR-4501), 3 up-regulated miRNAs (miR-371a-5p, miR-34c-5p and miR-1323) and 2 novel miRNAs. Additionally, corresponding alterations of pathways-correlated target mRNAs were observed including 5 up-regulated mRNAs (ICAM1, WNT2B, MYC, HLA-F and TGF-β1) and 3 down-regulated mRNAs (CDH1, PTENP1 and HSP90AA1).

Conclusions

Our study provides an overview of miRNA expression profile and the interactions between miRNA and their target mRNAs, which will deepen our understanding of the important roles of miRNAs in NPC radioresistance.     

2000-2015年中国PM2.5浓度时空分布特征及其城乡差异

近40年来,中国快速经济发展引发较为严重的大气污染,PM_2.5是一种重要的空气污染物,掌握其时空分布规律是对其进行防治的重要前提。基于遥感反演出的PM_2.5浓度数据集,研究了中国2000-2015年PM_2.5浓度的时空分布特征,并基于界定的1376个城镇城区及对应乡村的边界分析了每年PM_2.5浓度值的城乡差异,用线性趋势分析法计算城镇PM_2.5浓度的年际变化速率及显著性。结果表明,研究期内,PM_2.5浓度高于35 μg/m³的面积比例由18.58%增加至32.03%,低于15 μg/m³的面积从43.92%减少到25.12%。PM_2.5污染最严重的地区分布在塔里木盆地、河北南部、河南北部和山东西部。从2000年到2015年,中国绝大多数城镇PM_2.5浓度显著增加,尤其是在东北平原、太行山以东的河北省西南部、燕山以南的北京天津及河北唐山、鲁中南山地丘陵及周围平原地区、华北平原江苏省北部。PM_2.5城乡差异在河北省、山西省两条东北-西南向S形条带区域、浙江省-福建省条带及天山北部绿洲区域较大。研究对PM_2.5高浓度区域、PM_2.5浓度增长较快区域以及城区PM_2.5浓度对乡村影响较大区域进行图示,为中国进一步控制雾霾污染提供一定科学依据。     

沙质草地3种优势植物叶片光合生理对增温和降水减少的响应北大核心CSCD

以沙质草地优势物种猪毛蒿、胡枝子和糙隐子草为研究对象,利用开顶式生长室(OTC)模拟增温,研究降水减少20%、40%和60%与增温的交互作用对3种典型植物叶片光合气体交换特征及叶绿素荧光特征的影响,以揭示沙质草地3种优势植物对气候变化的响应规律。结果显示:(1)与自然温度相比,OTC模拟增温增加了猪毛蒿C_(i),显著降低了胡枝子G_(s)、P_(n)和T_(r)、糙隐子草G_(s)和P_(n)、猪毛蒿WUE和L_(s),也显著降低了猪毛蒿和胡枝子F_(v)/F_(m)和F_(v)/F_(o)。(2)无论增温与否,随着降水减少幅度的增加,猪毛蒿G_(s)和P_(n)呈下降趋势,且中度以上的干旱胁迫下(降水减少>40%)胡枝子和糙隐子草P_(n)显著低于对照。(3)在自然温度条件下,轻度干旱胁迫时(降水减少20%)猪毛蒿T_(r)显著低于对照,重度干旱胁迫时(降水减少60%)其WUE、F_(v)/F_(m)和F_(v)/F_(o)显著低于对照;重度干旱胁迫时,胡枝子C_(i)显著高于对照,差异幅度达10.7%,L_(s)显著低于对照,轻度干旱胁迫时(降水减少20%)其F_(v)/F_(m)和F_(v)/F_(o)显著高于中度以上的干旱胁迫;中度以上的干旱胁迫下糙隐子草T_(r)和G_(s)显著低于对照,重度干旱胁迫时,其C_(i)、F_(v)/F_(m)和F_(v)/F_(o)显著低于对照,WUE和L_(s)显著高于对照。(4)增温与降水减少交互作用下,所有处理猪毛蒿C_(i)均高于对照,差异幅度分别达4.5%,6.0%和8.4%;胡枝子T_(r)均显著低于对照,差异幅度达57.8%;重度干旱胁迫时猪毛蒿L_(s)和WUE显著低于对照,糙隐子草F_(v)/F_(m)和F_(v)/F_(o)随降水减少而降低,中度以上的干旱胁迫时其值显著低于对照。(5)相关性分析表明,3个优势物种的P_(n)与F_(v)/F_(m)和F_(v)/F_(o)均呈显著正相关关系,其中猪毛蒿和糙隐子草的P_(n)—F_(v)/F_(m)和P_(n)—F_(v)/F_(o)斜率明显高于胡枝子。研究表明,气候变暖会在一定程度上加剧降水减少对沙质草地3种群落优势物种光合作用的抑制。     

大鼠催乳素基因真核细胞可表达性质粒的构建及应用研究

７３５ｂｐ的ＰＲＬｃＤＮＡ片段从质粒ＰＲＬ－ＳＰ６５＃１中回收后,用粘性末端连接法将其重组到真核表达载体ｐｃＤＮＡ３上,筛选出正向连接重组体ｐｃＤＮＡ３－ＰＲＬＳ和反向连接重组体ｐｃＤＮＡ３－ＰＲＬＡＳ。将重组体ｐｃＤＮＡ３－ＰＲＬｓ和空载体ｐｃＤＮＡ３分别转入ＮＩＨ３Ｔ３细胞系,用Ｇ４１８筛选出阳性细胞后与未转染的ＮＩＨ３Ｔ３细胞在加Ｅ２和不加Ｅ２的情况下,用原位杂交的方法,分别用ＰＲＬｃＤＮＡ探针和原癌基因ｃ－Ｈ－ｒａｓｃＤＮＡ探针进行检测,未转染的ＮＩＨ３Ｔ３细胞在加Ｅ２和不加Ｅ２时都几乎无催乳素基因的表达,同样,转入空载体的ＮＩＨ３Ｔ３细胞也无ＰＲＬ的表达,而转入重组体ｐｃＤＮＡ３－ＰＲＬＳ的ＮＩＨ３Ｔ３细胞则有大量的ＰＲＬ基因的表达,与对照组相比有显著差异（Ｐ＜０．０１）。正常和转入空载体的ＮＩＨ３Ｔ３细胞有一定程度的原癌基因ｃ－Ｈ－ｒａｓ的表达,当分别加入Ｅ２和转入重组体ｐｃＤＮＡ３－ＰＲＬＳ后,ＮＩＨ３Ｔ３细胞中的ｃ－Ｈ－ｒａｓ基因表达水平都显著升高（Ｐ＜０．０５）。     

药用植物灯笼草的化学成分研究

从药用植物灯笼草(Clinopvdium polycephalum)中分离鉴定了5个化合物。其中主要成分为乌索酸(ursolic acid,Ⅰ),其余4个分别鉴定为:异樱花素(isosakuranetin.Ⅱ),香蜂草甙(didymin,Ⅲ),6'-十六碳酸酯基-α-菠甾醇-3-O-β-D-葡萄糖甙(6’-Palmityl-α-spinasteryl-3-O-β-D-glucoside,Ⅳ a)和十八碳酸酯基-α-波甾醇-3-O-β-D-葡萄糖甙(6’-stearyl-α-spinasteryl-3-O-β-D-glucoside,Ⅳ b)。上述化合物在该植物中均为首次报道。     

广西桉树人工林叶面积指数的估测及其校正

叶面积指数(Leaf area index, LAI)是森林生态系统重要的结构参数,通过遥感技术可反演区域LAI,但其可靠性需要地面准确的实测数据进行验证。选取广西国有高峰林场不同林龄的桉树(Eucalyptus robusta)人工林为对象,以异速生长法(Allometry)为对照,综合利用植物冠层分析仪法(LAI-2200)、跟踪辐射和冠层结构分析仪法(TRAC)、半球摄影法(DHP)以及地基激光雷达法(TLS)等间接法估测样地的LAI,并考虑木质成分以及聚集效应影响,进行相应的校正处理,为地面快速、准确测量桉树人工林LAI提供参考。结果表明：桉树人工林的比叶面积为125.37±13.38 cm~2/g,通过Allometry获得的LAI变化范围在1.65—3.84,平均为2.73,不同林龄间的差异均显著(P<0.05),随着林龄的增加呈现先增加后减少的趋势。在未校正情况下,LAI-2200、TRAC、DHP、TLS估算的LAI存在显著差异(P<0.05)。与对照相比,LAI-2200在幼龄林和过熟林中估算误差最小,TRAC在成熟林中估算误差最小。相对于完全去除法,利用...