Isolation and functional analysis of a 24-residue linear alpha-helical antimicrobial peptide from Korean blackish cicada, Cryptotympana dubia (Homoptera)

A new antimicrobial peptide, cryptonin, was isolated and characterized from the adult Korean blackish cicada, Cryptotympana dubia. It consists of 24 amino acid residues and has a molecular weight of 2,704 Da on mass spectroscopy. The predicted alpha-helical structure analysis and increased helix percent in 40% trifloroethanol of cryptonin suggests that it belongs to the typical linear alpha-helix forming peptide. Binding of the biotin-labeled cryptonin at the surface of E. coli cells and increased influx of propidium iodide in E. coli after cryptonin treatment indicates that it kills microbial cells by binding bacterial cell surfaces and disrupting the cell permeability. Cryptonin showed strong antibacterial (MIC 1.56-25 microg/ml) and antifungal (MIC 3.12-50 microg/ml) activities against tested bacteria and fungi including two antibiotic-resistant bacterial strains; methicilin-resistant S. aureus and vancomycin-resistant Enterococci (MIC 25 microg/ml, each).     

探讨模拟冷空气降温过程对健康大鼠和高血压大鼠凝血功能的影响

目的:通过模拟冷空气温度变化过程给予健康大鼠和高血压大鼠冷刺激,以此探讨冷空气过程对机体凝血功能的影响。方法:收集张掖市2011年3月一次典型冷空气过程数据,利用气象环境模拟箱模拟其温度变化过程。将24只雄性健康大鼠和24只雄性高血压大鼠分别随机分成最低温组(Tmin组)、Tmin对照组、复温组(Tr组)和Tr对照组。将Tmin组和Tr组大鼠放入气候箱中暴露冷空气温度变化过程。在Tmin和Tr时点分别停止Tmin组大鼠和Tr组大鼠冷空气暴露,并采血以测定其凝血功能指标—凝血四项。结果:与对照组相比,健康大鼠和高血压大鼠活化部分凝血酶原时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)在降温后与对照组相比没有显著差异(P>0.05),但血浆纤维蛋白原(Fbg)含量在高血压大鼠和健康大鼠Tmin组明显高于其对照组(P<0.01)。温度恢复后,其在健康组含量仍然高于其对照组(P<0.05),而在高血压大鼠中没有差异(P>0.05)。同Fbg,反应Fbg的纤维蛋白原时间(Fbg-time)在健康大鼠Tmin和Tr组中短于对照组(P<0.01,),而在高血压大鼠中仅在Tmin组短于对照组(P<0.01)。高血压大鼠血中的Fbg含量和Fbgt明显高于和短于健康大鼠(P<0.01)。结论:①冷空气降温过程能增加机体血中Fbg含量,使凝血功能增强,可能增加心血管疾病危险性;②冷空气刺激对健康大鼠凝血功能影响强于高血压大鼠。     

镉诱导金属硫蛋白在华溪蟹组织中的表达

采用体外暴露染毒法,研究了不同浓度与时间条件下,镉诱导河南华溪蟹(Sinopotamon henanense)金属硫蛋白(metallothionein,MT)在肝胰腺、肌肉、鳃和卵巢中的表达差异。镉浓度分别为0、14·5mg/L、29mg/L和58mg/L;处理时间依次为1d、3d和5d。利用镉血红蛋白饱和法和火焰原子吸收分光光度法(AAS)测定MT的蛋白含量。结果显示,用不同的染毒浓度和处理时间,镉在组织中诱导产生MT的含量有较大差异,其中肝胰腺MT的诱导量最大,变化规律也最明显;肌肉中也有较大量MT的表达;而鳃和卵巢MT的诱导量均较低。此外,本文分析了镉的浓度与时间梯度对诱导MT表达的影响与毒性效应机制。结论:组织不同,染毒浓度及时间不同,镉诱导MT的表达也不同,具有一定的组织差异性和规律性。     

利用癌功能类从癌基因组突变谱中识别癌基因

从大规模癌样本基因突变扫查数据中识别癌基因具有重要的意义. 一些重要功能的改变对于癌的发生发展是必需的, 因此将它们定义为癌功能类, 并从GO(Gene Ontology)中选择一组显著富集已知癌基因的细致功能类来代表它们. 为了评价以癌相关功能类作为特征识别癌基因的效果, 将已知的蛋白激酶癌基因定义为阳性金标准, 而将其他的蛋白激酶基因定义为阴性金标准. 结果表明, 与利用选择压力作为特征的方法比较, 利用癌相关功能类作为特征的方法可以更有效地识别癌基因. 进一步结合癌相关功能类与基因非同义突变个数可以产生更可靠的预测结果. 最后, 将46个注释到癌相关功能类并且其非同义突变个数至少为3的蛋白激酶基因预测为癌基因, 预测精确率达到0.42.     

新疆维吾尔族妇女宫颈癌组织中乳头瘤病毒16型E6基因的克隆和序列分析

为了分析新疆南部地区维吾尔族妇女宫颈癌组织中HPV16型E6基因结构特点，从中国新疆南部地区维吾尔族妇女宫颈癌活检组织标本中提取DNA，以宫颈癌活检组织标本DNA为模板进行PCR扩增，获得HPV16 E6基因，将其克隆到pUCm-T载体上，并对其进行基因全序列分析.PCR检测结果显示宫颈癌组织中HPV16 E6阳性率为82.35%（14/17）；测序结果显示，新疆株HPV16 E6基因全长456 bp，大小与德国标准株一致.E6基因的第247位碱基发生T→G突变，并由此引起所编码的氨基酸亦发生改变.上述结果表明，中国新疆南部地区维吾尔族妇女宫颈癌患者组织中HPV16 E6的基因结构与德国标准株HPV16 E6基因之间存在差异.     

MRI Features in a Canine Model of Ischemic Stroke: Correlation between Lesion Volume and Neurobehavioral Status during the Subacute Stage

The purpose of this study was to evaluate the diagnostic value of magnetic resonance imaging (MRI) and assess the correlation between the volume of the ischemic lesion and neurobehavioral status during the subacute stage of ischemic stroke. Ischemic stroke was induced in 6 healthy laboratory beagles through permanent occlusion of the middle cerebral artery (MCAO). T2-weighted and fluid-attenuated inversion recovery (FLAIR) imaging, diffusion-weighted imaging (DWI), measurement of the apparent diffusion coefficient (ADC) ratio, and neurobehavioral evaluation were performed 3 times serially by using a 1.5-T MR system: before and 3 and 10 d after MCAO. Ischemic lesions demonstrated T2 hyperintensity, FLAIR hyperintensity, and DWI hyperintensity. The ADC ratio was decreased initially but then was increased at 10 d after MCAO. Ischemic lesion volumes on T2-weighted and FLAIR imaging were not significantly different from those on DWI. The lesion volume and neurobehavioral score showed strong correlation. Our results suggest that conventional MRI may be a reliable diagnostic tool during the subacute stage of canine ischemic stroke.Abbreviations: ADC, apparent diffusion coefficient; DWI, diffusion-weighted imaging; FLAIR, fluid-attenuated inversion recovery; MCAO, middle cerebral artery occlusion; MRI, magnetic resonance imaging; PWI, perfusion-weighted imagingIn human medicine, stroke is a leading cause of adult mortality and neurologic disability worldwide.¹ Strokes previously were thought to be uncommon in small animals, but the true prevalence is unknown.⁴ These events are now recognized more frequently in dogs because of increased use of magnetic resonance imaging (MRI).^5,14,17Because the infusion of thrombolytic agents, such as urokinase or tissue plasminogen activator, within 3 to 6 h of the onset symptoms is effective in restoring blood flow and improving stroke outcome in humans,¹⁹ the detection of early ischemic changes is now thought to be necessary to improve patient outcome. Computed tomography and conventional MRI are not sufficiently sensitive to predict the presence and extent of ischemic damage during the acute stage after a stroke.^12,20 Therefore several MRI sequences, such as fluid-attenuated inversion recovery (FLAIR), diffusion-weighted imaging (DWI), perfusion-weighted imaging (PWI), and MR angiography, have been developed for early diagnosis and subsequent follow-up of ischemic stroke.³ High-field magnetic strengths (at least 1.5 T) are necessary to perform these sequences.In contrast to the situation in humans, ischemic stroke in many dogs is diagnosed during the subacute stage—24 h to 6 wk after the vascular insult—due to the time lag between the onset of clinical signs to referral and to the lack of standard diagnostic protocols for ischemic stroke in dogs. In most reports of strokes in dogs, the median interval between the onset of neurologic dysfunction and performance of an MRI was more than 2 d.^5,14,17 Whereas DWI has marked sensitivity to very early ischemic changes in the brain, T2-weighted and FLAIR images gradually become more hyperintense later (that is, during the first 24 h after the insult).³ Therefore, hyperintensity on T2-weighted and FLAIR images is believed to be representative of mature lesions.¹⁵ In light of these findings, we hypothesized that conventional MR sequences, such as T2-weighted and FLAIR imaging as well as DWI would be used for the diagnosis of the subacute stage of ischemic stroke in dogs.The purpose of this study was to evaluate the diagnostic value of MRI and assess the correlation between the volume of ischemic lesions and neurobehavioral status during the subacute stage of ischemic stroke in dogs. We therefore investigated the lesion volume of T2-weighted and FLAIR images compared with that on DWI images. Furthermore, we assessed the relationship between the apparent diffusion coefficient (ADC) of the ischemic lesions and the neurobehavioral status of the dogs.     

野艾蒿挥发油对HeLa癌细胞形态与结构的影响

采用MTT法检测细胞活力,用倒置显微镜、荧光显微镜和扫描电子显微镜观察细胞形态与结构的变化,用激光共聚焦显微镜观察细胞微管的分布,从而研究了野艾蒿挥发油对HeLa人宫颈癌细胞形态与结构的影响。结果表明:(1)野艾蒿挥发油对HeLa癌细胞的增殖有明显的抑制作用,呈剂量和时间依赖性。(2)野艾蒿挥发油处理HeLa癌细胞24h后,100、200μg/mL实验组细胞体积缩小,核染色质凝集、微绒毛消失、细胞表面有泡状突起,微管解聚,呈现典型的凋亡特征;400μg/mL实验组细胞膜破裂、胞浆内含物外泄,呈明显的坏死特征。(3)野艾蒿挥发油具有抑制HeLa癌细胞增殖的作用,低、中浓度的野艾蒿挥发油诱导细胞凋亡,而高浓度的野艾蒿挥发油引起细胞坏死。     

Relationship of the lung microbiome with PD-L1 expression and immunotherapy response in lung cancer

IL-35 subunit EBI3 is up-regulated in pulmonary fibrosis tissues. In this study, we investigated the pathological role of EBI3 in pulmonary fibrosis and dissected the underlying molecular mechanism. Bleomycin-induced pulmonary fibrosis mouse model was established, and samples were performed gene expression analyses through RNAseq, qRT-PCR and Western blot. Wild type and EBI3 knockout mice were exposed to bleomycin to investigate the pathological role of IL-35, via lung function and gene expression analyses. Primary lung epithelial cells were used to dissect the regulatory mechanism of EBI3 on STAT1/STAT4 and STAT3. IL-35 was elevated in both human and mouse with pulmonary fibrosis. EBI3 knockdown aggravated the symptoms of pulmonary fibrosis in mice. EBI3 deficiency enhanced the expressions of fibrotic and extracellular matrix-associated genes. Mechanistically, IL-35 activated STAT1 and STAT4, which in turn suppressed DNA enrichment of STAT3 and inhibited the fibrosis process. IL-35 might be one of the potential therapeutic targets for bleomycin-induced pulmonary fibrosis.     

壶腹周围憩室对胆总管结石患者结石成分及胆道菌群的影响

探究壶腹周围憩室(periampullary diverticulum, PAD)对胆总管结石成分及胆道菌群的影响。

收集我院行内镜下取石治疗患者44例, 其中14例伴有PAD、30例无PAD, 使用红外光谱法和分光光度法分析结石样本中胆固醇、胆红素含量。从入组患者中分别筛选8例伴有PAD的患者为PAD组, 8例无PAD的为对照组, 收集患者胆汁样本, 提取样本中菌群DNA进行扩增并构建细菌基因文库, 经Illumina MiSeq平台高通量测序后进行生物信息学分析。最后对差异菌群与胆红素、胆固醇进行相关性分析。

2组患者对比年龄、胆管直径、结石直径和总胆红素(TBIL)等资料差异均无统计学意义(均P > 0.05)。2组患者结石中胆固醇和胆红素含量[(39.77±13.00)%vs(61.80±26.00)%, Z=-2.973, P=0.003;(27.03±13.00)%vs(12.27±16.00)%, Z=-2.999, P=0.003)差异存在统计学意义。PAD组与对照组Alpha多样性差异无统计学意义(P > 0.05), 而在物种群落组成上存在差异。卟啉单胞菌属、嗜胆菌属等10种菌属在PAD组中的丰度高于对照组, 真杆菌属、乳杆菌属较对照组降低, 差异均有统计学意义(均P < 0.05)。相较于对照组, PAD组牛磺酸和次牛磺酸的代谢、钙信号通路等5条通路差异存在统计学意义(均P < 0.05)。嗜胆菌属和卟啉单胞菌属与胆红素具有相关性(R²=0.701 3, P < 0.000 1;R²=0.286 8, P=0.032 5)。

伴有PAD的患者结石含有更高的胆红素成分。PAD的存在改变胆道菌群多样性、构成及丰度, 并可能通过菌群变化影响结石成分。

     

野生与人工养殖鲫鳃部寄生指环虫的群落结构

为了阐明养殖环境对鱼类寄生指环虫群落的影响,研究调查了湖北省梁子湖和湖北省石首市养殖池塘中鲫鳃部寄生指环虫的种类和感染情况。在梁子湖中的野生鲫共发现6种指环虫,分别为坏鳃指环虫（Dactylogyrus vastator）、中型指环虫（D. intermedius）、弧形指环虫（D. arcuatus）、弓茎指环虫（D. baueri）、美丽指环虫（D. formosus）、望外指环虫（D. inexpeatatus）;在养殖池塘中,除了望外指环虫外,其他5种指环虫均有发现。分析结果表明,来自于梁子湖野生鲫鳃部的坏鳃指环虫的平均丰度以及其他4种指环虫（不含望外指环虫）的感染率与平均丰度明显高于养殖池塘的。梁子湖中的指环虫群落平均物种丰富度、平均寄生虫数量以及Brillouin多样性指数都明显高于养殖池塘的;梁子湖野生鲫寄生指环虫的优势种是弧形指环虫（0.54）,而池塘中的是坏鳃指环虫（0.76）。在宿主密度、种群大小、光照和溶氧等影响指环虫群落的因子中,溶氧可能是最主要因子。