The tetraspanin CD9 associates with transmembrane TGF-alpha and regulates TGF-alpha-induced EGF receptor activation and cell proliferation

Transforming growth factor-alpha (TGF-alpha) is a member of the EGF growth factor family. Both transmembrane TGF-alpha and the proteolytically released soluble TGF-alpha can bind to the EGF/TGF-alpha tyrosine kinase receptor (EGFR) and activate the EGFR-induced signaling pathways. We now demonstrate that transmembrane TGF-alpha physically interacts with CD9, a protein with four membrane spanning domains that is frequently coexpressed with TGF-alpha in carcinomas. This interaction was mediated through the extracellular domain of transmembrane TGF-alpha. CD9 expression strongly decreased the growth factor- and PMA- induced proteolytic conversions of transmembrane to soluble TGF-alpha and strongly enhanced the TGF- alpha-induced EGFR activation, presumably in conjunction with increased expression of transmembrane TGF-alpha. In juxtacrine assays, the CD9-induced EGFR hyperactivation by transmembrane TGF-alpha resulted in increased proliferation. In contrast, CD9 coexpression with transmembrane TGF-alpha decreased the autocrine growth stimulatory effect of TGF-alpha in epithelial cells. This decrease was associated with increased expression of the cdk inhibitor, p21(CIP1). These data reveal that the association of CD9 with transmembrane TGF-alpha regulates ligand-induced activation of the EGFR, and results in altered cell proliferation.     

建立ICU呼吸专科护理小组在机械通气病人中的作用

目的:探讨建立ICU呼吸专科护理小组在机械通气病人中的作用。方法:评估建立ICU呼吸专科护理小组前后实施呼吸机集束策略依从性的情况及对呼吸机相关性肺炎(VAP)发生率的影响。结果:建立ICU呼吸专科护理小组前后分别纳入458、487个病人,手卫生依从性由20.9%提高到65.7%(P=0.001);床头抬高30°以上依从性由24.8%提高到49.5%(P=0.001);镇静评分依从性无明显改善(P=0.06);使用无创呼吸机依从性由20.4%提高到32%(P=0.03)。同时ICU呼吸专科护理小组建立后可降低VAP发生率(24.4%vs 41.7%,P=0.004),降低VAP病人的机械通气时间和ICU住院时间,但并不能降低其死亡率。结论:建立ICU呼吸专科护理小组可提高呼吸机集束策略的依从性,可以降低VAP发生率。     

NF-κB激活gp96转录从而促进肝细胞生长、细胞周期进展和细胞转化

【目的】探讨乙肝感染中gp96上调的机制及其可能发挥的病理学机制。【方法】首先通过生物信息学、Real-time PCR、荧光报告基因和Western blot研究NF-κB激活gp96表达的机制。进一步通过在肝细胞中过表达或敲低gp96的水平,运用CCK-8法和流式检测分析gp96对肝细胞增殖、凋亡,和细胞周期的影响,通过检测肝细胞EMT发生和细胞集落形成实验,分析gp96对于HCC发生的作用。【结果】NF-κB与gp96启动子上NF-κB结合位点结合,激活gp96的表达。实验结果显示,gp96能够促进肝细胞增殖、抑制凋亡,促进细胞周期从静息期向分裂期的转化,同时促进肝细胞EMT发生和细胞集落的形成。【结论】NF-κB通过活化gp96启动子上调其表达,为HBV慢性感染上调gp96的机制提供了线索,同时提示gp96在慢性炎症引发HCC过程中发挥重要作用。     

The strain HEB01 of Fusarium sp., a new pathogen that infects brown soft scale

The objectives of the present study were to identify a fungal strain, HEB01, isolated from naturally infected brown soft scale, Coccus hesperidum L. (Hemiptera: Coccidae), and to determine whether it is an entomopathogenic fungus. Fungal culture, reinoculation test, morphological observations, and infection behaviors were investigated. Additionally, the fungal gene sequence of translation elongation factor 1-a (EF-1a) was obtained for molecular identification. The results showed that the fungal strain HEB01 belongs to the Fusarium incarnatum-equiseti species complex and is part of the family Nectriaceae (Hypocreales: Sordariomycetes). The inoculation test and observations of infection behaviors indicated that strain HEB01 is a pathogenic fungus and confirmed that it infects brown soft scale. Thus, the HEB01 strain of Fusarium incarnatum-equiseti is the first pathogen in the genus Fusarium to be isolated from a brown soft scale.     

Estrogen improves the proliferation and differentiation of hBMSCs derived from postmenopausal osteoporosis through notch signaling pathway

Estrogen deficiency is the main reason of bone loss, leading to postmenopausal osteoporosis, and estrogen replacement therapy (ERT) has been demonstrated to protect bone loss efficiently. Notch signaling controls proliferation and differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Moreover, imperfect estrogen-responsive elements (EREs) were found in the 5′-untranslated region of Notch1 and Jagged1. Thus, we examined the molecular and biological links between estrogen and the Notch signaling in postmenopausal osteoporosis in vitro. hBMSCs were obtained from healthy women and patients with postmenopausal osteoporosis. Notch signaling molecules were quantified using real-time polymerase chain reaction (real-time PCR) and Western Blot. Luciferase reporter constructs with putative EREs were transfected into hBMSCs and analyzed. hBMSCs were transduced with lentiviral vectors containing human Notch1 intracellular domain (NICD1). We also used N-[N-(3, 5-diflurophenylacetate)-l-alanyl]-(S)-phenylglycine t-butyl ester, a γ-secretase inhibitor, to suppress the Notch signaling. We found that estrogen enhanced the Notch signaling in hBMSCs by promoting the expression of Jagged1. hBMSCs cultured with estrogen resulted in the up-regulation of Notch signaling and increased proliferation and differentiation. Enhanced Notch signaling could enhance the proliferation and differentiation of hBMSCs from patients with postmenopausal osteoporosis (OP-hBMSCs). Our results demonstrated that estrogen preserved bone mass partly by activating the Notch signaling. Because long-term ERT has been associated with several side effects, the Notch signaling could be a potential target for treating postmenopausal osteoporosis.     

混合碳源发酵生产头孢菌素C过程优化

本文对头孢菌素C（Cephalosporin C,CPC）发酵过程中碳源补料控制策略进行了优化研究,提出了一种基于DO—Stat的混合碳源流加策略,提高了发酵整体性能。在7L发酵罐上对使用该策略和传统补油策略的头孢菌素发酵性能进行比较,结果表明,采用补加混合碳源（葡萄糖＋豆油）策略时,CPC终浓度最高,达到36．99g／L,CPC得率也从使用传统单纯补油策略时的11．39％提高到22．19％,代谢副产物去乙酰氧头孢菌素C（DAOC）的积累量少,DAOC／CPC只有0．38％,达到生产要求。     

河北省华北落叶松河北群体遗传多样性分析

以来自河北8个群体共312个华北落叶松天然个体为材料,利用10对SSR引物对群体进行扩增,所得数据用于其遗传多样性和遗传结构的研究。结果表明:10对SSR位点共检测到42个位点,平均每个位点有4.2个等位基因,位点的等位基因数(Na)为2~6个。群体平均等位基因数为3.36,Shannon指数(I)平均为0.749,观察杂合度Ho为0.405、期望杂合度He为0.423,来自河北北部的围场台子水和吉字以及兴隆雾灵山的多样性水平最高,南部的阜平群体多样性水平最低。AMOVA分子差异分析显示3%的遗传变异存在于群体间,97%的遗传变异则存在于群体内,基因分化系数Gst仅为0.028,表明华北落叶松在河北的群体分化很低。聚类结果表明河北围场两个群体为一类,蔚县、隆化和涿鹿为一类,阜平和赤城为一类,兴隆雾灵山群体距离其他较远。多样性北部群体较高,南部较低,表明华北落叶松可能的衍化方向是从北向南,印证了北部可能是起源中心的结论。     

线粒体基因组D-Loop区基因多态性与2型糖尿病的相关性

随机选取199例浙江地区2型糖尿病患者与102例正常对照, 采用聚合酶链反应(Polymerase chain re-action, PCR)、基因片段直接测序来检测线粒体基因组D-Loop区域基因变异情况, 同时分析其与主要临床指标的关系。结果显示: 线粒体基因组D-Loop区域为一高变异区, np73A-G、np263A-G、np16223C-T、np16519T-C为4个高变异位点; 发现29个未见报道的新变异位点; np193A-G、np234A-G、np16108C-T等变异与糖尿病家族史有关。这表明浙江籍汉族人线粒体基因组D-Loop区存在大量基因多态性现象, 此区域的某些变异可能与糖尿病的发生发展等具有一定相关性。     

Molecular phylogeny and taxonomic reconsideration of the subfamily Zapodinae (Rodentia: Dipodidae), with an emphasis on Chinese species

Although the subfamily Zapodinae (Rodentia, Dipodidae) contains only five species, the phylogeny and taxonomy of these species are still being disputed. First, whether Eozapus and Napaeozapus should be treated as independent genera or subgenera of Zapus has been argued for a long period. Second, the subspecific genetic differentiation of Chinese jumping mouse (Eozapus setchuanus) has not been studied in detail, neither from morphological nor molecular aspects. In this study, the phylogenetic relationship among all the five species of Zapodinae was reconstructed using DNA sequence data from the mitochondrial cytochrome b gene and the nuclear interphotoreceptor retinoid binding protein gene. Bayesian inference, maximum parsimony and maximum likelihood analyses were conducted. The results showed that two major clades could be recognized within Zapodinae. Eozapus setchuanus, is the species endemic to China, strongly formed a monophyletic clade. In the other clade, genus Zapus received significant support in all analyses to be the sister group of the genus Napaeozapus. By comparing genetic distances among these three genera, we conclude that both Eozapus and Napaeozapus should be considered as valid genera rather than subgenera of Zapus. Furthermore, we observed that the two subspecies of E. setchuanus did not form reciprocally monophyletic groups, thus the traditional taxonomy which divided E. setchuanus into two subspecies based on only one morphological character was questionable.     

高羊茅胚性悬浮细胞系的快速建立及其单细胞培养

本研究采用高羊茅(Millennium和Hundog Ⅴ)成熟种子为材料,以MS为基本培养基,通过添加2.5 mg/L CuSO4·5H2O,或提高NH4NO3浓度至2.5 g/L等措施均能明显提高愈伤组织的质量,经过3～5个月的筛选获得疏松干燥、颗粒状、生长旺盛、适合悬浮培养的Ⅱ型胚性愈伤组织.悬浮培养初期需用MSⅠ液体培养基进行一个月的启动培养,之后转用MSⅡ继代保持,约2个月左右即建立起来自高羊茅2个品种的3个悬浮细胞系.生长特性测定结果表明,悬浮培养的初始接种量以1.0～3.0 ml/40 ml为宜,生长周期内其pH值不断波动变化,最适范围在5.0～5.7之间.5～8 d为悬浮系的对数生长期,此时细胞分裂旺盛,增殖较快,是分离单细胞的最佳时期.单细胞培养方式以悬浮培养效果最好.