mTOR and differential activation of mitochondria orchestrate neutrophil chemotaxis

Neutrophils use chemotaxis to locate invading bacteria. Adenosine triphosphate (ATP) release and autocrine purinergic signaling via P2Y2 receptors at the front and A2a receptors at the back of cells regulate chemotaxis. Here, we examined the intracellular mechanisms that control these opposing signaling mechanisms. We found that mitochondria deliver ATP that stimulates P2Y2 receptors in response to chemotactic cues, and that P2Y2 receptors promote mTOR signaling, which augments mitochondrial activity near the front of cells. Blocking mTOR signaling with rapamycin or PP242 or mitochondrial ATP production (e.g., with CCCP) reduced mitochondrial Ca²⁺ uptake and membrane potential, and impaired cellular ATP release and neutrophil chemotaxis. Autocrine stimulation of A2a receptors causes cyclic adenosine monophosphate accumulation at the back of cells, which inhibits mTOR signaling and mitochondrial activity, resulting in uropod retraction. We conclude that mitochondrial, purinergic, and mTOR signaling regulates neutrophil chemotaxis and may be a pharmacological target in inflammatory diseases.     

副干酪乳杆菌HD1.7群体感应行为

【目的】Paracin1.7是从副干酪乳杆菌(Lactobacillus paracasei)HD1.7发酵液中提取的一种细菌素,本文主要研究菌株HD1.7在发酵过程中调控Paracin1.7代谢的群体感应机制。【方法】利用杯碟法检测不同生长条件下菌株HD1.7培养液的抑菌活性,通过调整培养基营养成分的多寡,控制培养液中细胞密度。【结果】菌株HD1.7的抑菌活性与其细胞密度密切相关,只有当细胞密度达到一定的阈值(OD600为0.8,菌体干重为0.331 1 g/L)时,菌株才能表现抑菌活性;以发酵上清液作为信号分子,当添加不同浓度信号分子至低于阈值浓度培养液后,菌株抑菌活性受到不同程度的影响,并且在去除信号分子后,菌株的抑菌活性明显降低。【结论】细菌素Paracin1.7是存在于HD1.7发酵液中的特殊的群体感应信号分子,可进行自我诱导。细菌素Paracin1.7的抑菌活性受到HD1.7群体感应系统的调控。     

The Effect of Sulfur on the Composition of Arbuscular Mycorrhizal Fungal Communities During the Pod-Setting Stage of Different Soybean Cultivars

This study sought to investigate the effect of sulfur levels on changes in the fungal community composition of arbuscular mycorrhizae (AM) at the pod-setting stage and the relationship between the amount of applied sulfur and AM fungal diversity in different soybean cultivars. The objective of the research was to determine the optimal sulfur application level for different soybean cultivars and to improve soybean yield and quality from the perspective of AM fungal diversity. Three soybean cultivars, Heinong 44, Heinong 48, and Heinong 37, were selected as study materials. In addition to 0.033?g each of N, P₂O₅ and K₂O per kg of soil, 0, 0.02, 0.04, or 0.06?g of elemental sulfur was applied to each kg of soil for the four treatment groups, S1, S2, S3, and S4, respectively. The AM fungal community structure was analyzed in the soil and root of different soybean cultivars using the PCR-DGGE technology. The results indicated a significant effect of sulfur on the AM fungal community structure in the roots and rhizospheric soil of different soybean cultivars. The three soybean cultivars in group S2 exhibited the highest diversity in AM fungus. Significant changes in the dominant fungal species were observed in the DGGE fingerprints of each sample, and Glomus, Funneliformis, Rhizophagus, and Claroideoglomus fungi were the dominant species of AM fungus in the roots and soil of soybean. The application of an appropriate amount of sulfur improved the diversity of AM fungi in roots and rhizospheric soil of different soybean cultivars.     

HnRNPL as a key factor in spermatogenesis: Lesson from functional proteomic studies of azoospermia patients with sertoli cell only syndrome

Sertoli cell only syndrome (SCOS) is one of the main causes leading to the abnormal spermatogenesis. However, the mechanisms for abnormal spermatogenesis in SCOS are still unclear. Here, we analyzed the clinical testis samples of SCOS patients by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF/TOF MS) to find the key factors contributing to SCOS. Thirteen differential proteins were identified in clinical testis samples between normal spermatogenesis group and SCOS group. Interestingly, in these differential proteins, Heterogeneous nuclear ribonucleoprotein L(HnRNPL) was suggested as a key regulator involved in apoptosis, death and growth of spermatogenic cells by String and Pubgene bioinformatic programs. Down-regulated HnRNPL in testis samples of SCOS patients was further confirmed by immunohistochemical staining and western blotting. Moreover, in vitro and in vivo experiments demonstrated that knockdown of HnRNPL led to inhibited proliferation, increased apoptosis of spermatogenic cell but decreased apoptosis of sertoli cells. Expression of carcinoembryonic antigen-related cell adhesion molecule 1 in GC-1 cells or expression of inducible nitric oxide synthases in TM4 sertoli cells, was found to be regulated by HnRNPL. Our study first shows HnRNPL as a key factor involved in the spermatogenesis by functional proteomic studies of azoospermia patients with sertoli cell only syndrome. This article is part of a Special Issue entitled: Proteomics: The clinical link.     

Etiologic diagnosis of lower respiratory tract bacterial infections using sputum samples and quantitative loop-mediated isothermal amplification

Etiologic diagnoses of lower respiratory tract infections (LRTI) have been relying primarily on bacterial cultures that often fail to return useful results in time. Although DNA-based assays are more sensitive than bacterial cultures in detecting pathogens, the molecular results are often inconsistent and challenged by doubts on false positives, such as those due to system- and environment-derived contaminations. Here we report a nationwide cohort study on 2986 suspected LRTI patients across P. R. China. We compared the performance of a DNA-based assay qLAMP (quantitative Loop-mediated isothermal AMPlification) with that of standard bacterial cultures in detecting a panel of eight common respiratory bacterial pathogens from sputum samples. Our qLAMP assay detects the panel of pathogens in 1047(69.28%) patients from 1533 qualified patients at the end. We found that the bacterial titer quantified based on qLAMP is a predictor of probability that the bacterium in the sample can be detected in culture assay. The relatedness of the two assays fits a logistic regression curve. We used a piecewise linear function to define breakpoints where latent pathogen abruptly change its competitive relationship with others in the panel. These breakpoints, where pathogens start to propagate abnormally, are used as cutoffs to eliminate the influence of contaminations from normal flora. With help of the cutoffs derived from statistical analysis, we are able to identify causative pathogens in 750 (48.92%) patients from qualified patients. In conclusion, qLAMP is a reliable method in quantifying bacterial titer. Despite the fact that there are always latent bacteria contaminated in sputum samples, we can identify causative pathogens based on cutoffs derived from statistical analysis of competitive relationship. TRIAL REGISTRATION: ClinicalTrials.gov NCT00567827.     

Insulators, long-range interactions, and genome function

Insulators are DNA-protein complexes that can mediate interactions in cis or trans between different regions of the genome. Although originally defined on the basis of their ability to block enhancer-promoter communication or to serve as barriers against the spreading of heterochromatin in reporter systems, recent information suggests that their function is more nuanced and depends on the nature of the sequences brought together by contacts between specific insulator sites. Here we provide an overview of new evidence that has uncovered a wide range of functions for these sequences in addition to their two classical roles.     

心血管危重症继发交感风暴37例临床分析

目的:探讨心血管危重症继发交感风暴的病因分布特点和治疗方法。方法:回顾性分析我院2001年-2011年37例心血管危重症继发交感风暴临床资料。结果:37例心血管危重症继发交感风暴患者,其中男性22例,女性15例,年龄55-82岁,平均68.25岁。病因分布为:心肌梗死29例,心衰3例,扩张性心肌病2例,电解质紊乱2例,Brugada综合征1例。治疗方法包括β受体阻滞剂、胺碘酮、利多卡因、电除颤等。其中,17例因反复发作室颤,经抗心律失常药物和电除颤治疗无效死亡,余20例病情得到控制。结论:心血管危重症继发交感风暴临床病因多样,病情凶险,应用β受体阻滞剂及抗心律失常药物,同时积极针对病因及诱因治疗,可以改善临床症状和预后。     

Na2CO3胁迫下甜菜幼苗根际土壤环境因子的变化及其相关性

以两个甜菜品种‘KWS0143’(耐盐碱性强)和‘Beta464’(耐盐碱性较弱)为对象,设置4个Na₂CO₃浓度处理[占土壤质量的0%(CK)、0.4%、0.8%和1.2%],采用盆栽方法研究甜菜幼苗时期植株干质量、根际土壤酶活性和微生物数量的变化.结果表明: 与对照相比,0.4%处理的植株干质量明显增加,而0.8%和1.2%处理显著受到抑制,且处理间差异显著. 不同处理下甜菜幼苗根际土壤脲酶、过氧化氢酶和碱性磷酸酶活性表现出相似的变化规律,0.4%处理酶活性较对照有所增加但不显著;0.8%和1.2%处理则显著降低了酶活性,同时‘KWS0143’的土壤酶活性均高于‘Beta464’.与对照相比,0.4%处理土壤微生物群落没有显著变化;0.8%和1.2%处理的根际土壤细菌、真菌和放线菌数量显著减少,且‘KWS0143’的根际微生物数量高于‘Beta464’.两品种植株干质量、土壤酶和土壤微生物之间呈显著正相关;通径分析表明,‘KWS0143’植株干质量决定系数表现为:放线菌>细菌>过氧化氢酶>脲酶>真菌>碱性磷酸酶,‘Beta464’表现为:放线菌>过氧化氢酶>脲酶>真菌>碱性磷酸酶>细菌.     

乳腺透明细胞癌:一种罕见的乳腺癌亚型

目的探讨乳腺透明细胞癌(glycogen-rich clear cell carcinoma of breast,GRCCC)的临床病理特征和诊断要点。方法报道1例GRCCC的临床病理、免疫组织化学结果及预后,并复习相关文献。结果患者,女性,44岁,2016年7月因发现右乳无痛性包块3年余,于外院行右乳改良切除术,病检提示乳腺癌,右乳癌术后化疗后6月余,发现全身多发骨转移,后来我院会诊。镜下显示乳腺浸润性癌,细胞排列成巢索状,间质为毛细血管,肿瘤细胞具有丰富透明的胞浆,呈多边形,边界清楚,核深染,核仁明显,肿瘤伴大片坏死,可见多处淋巴管及血管内癌栓。免疫组织化学染色显示肿瘤呈ER、PR和Her-2三重阴性,CK5/6局灶阳性,GATA-3阳性,P120细胞膜阳性,E-cadherin阳性,标记血管的CD34阳性,标记淋巴管的D2-40阳性,GCDFP-15阴性。会诊结果诊断为乳腺透明细胞癌。最初的局部治疗6个月后,病人出现多发骨和远处淋巴结转移。结论乳腺富糖原的透明细胞癌是乳腺癌一种罕见的组织学亚型,通常预后较差。     

外源MeJA对镉胁迫下波斯菊生长及抗氧化系统的影响

以波斯菊种子和幼苗为试验材料,采用滤纸上发芽和叶面喷施盆栽幼苗的方法,探讨不同浓度(0、0.5、1.0、5.0、10.0和25.0μmol·L~(-1))茉莉酸甲酯(MeJA)对镉(100μmol·L~(-1) Cd~(2+))胁迫下波斯菊种子萌发、幼苗生长以及抗氧化系统的影响。结果表明,镉胁迫条件下,波斯菊的生长受到严重抑制,施加低浓度MeJA(1.0～5.0μmol·L~(-1))可以促进波斯菊种子萌发,增加波斯菊幼苗叶绿素含量,促进叶片干物质含量的积累和叶片相对含水量增加,提高叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)的活性,增加氧化型谷胱甘肽(GSH)含量和氧化型谷胱甘肽/还原型谷胱甘肽(GSH/GSSG)比值,降低游离脯氨酸(Pro)和丙二醛(MDA)含量;而高浓度的MeJA(大于10μmol·L~(-1))则表现出相反的抑制作用,影响镉胁迫下波斯菊种子萌发和幼苗的相关代谢。研究发现,适宜浓度的外源MeJA,在镉胁迫条件下可通过降低渗透调节物质的含量,诱导增强抗氧化酶活性,增加氧化型谷胱甘肽(GSH)含量,调整GSH/GSSG比例等途径促进波斯菊种子萌发和幼苗的生长,有效缓解镉胁迫对波斯菊的伤害,从而提高波斯菊幼苗的耐镉性能。