Advanced glycosylation end products induce nitric oxide synthase expression in C6 glioma cells: involvement of a p38 MAP kinase-dependent mechanism.

The mitogen-activated protein kinase (MAPK) pathway is believed to function as an important mediator of inducible nitric oxide synthase (iNOS) expression. In the present study, we investigated the role of the p38 MAPK signaling pathway in advanced glycosylation end products (AGEs)-induced iNOS expression in C6 glioma cells. AGEs caused a dose-dependent increase of nitrite accumulation in C6 glioma cells. The AGEs-stimulated nitrite production from C6 glioma cells was inhibited by actinomycin D, cyclohexamide, and the NO synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME), suggesting that the increase of AGEs-induced nitrite release is due to iNOS up-regulation. Consistently, treatment of C6 glioma cells with AGEs induced iNOS protein expression. AGEs-stimulated nitrite production was inhibited by pretreatment of C6 glioma cells with anti-AGEs antibodies (1:100 or 1:50). The tyrosine kinase inhibitor (genistein and tyrphostin), the Ras-farnesyl transferase inhibitor (FPT inhibitor-II), or the p38 MAPK inhibitor (SB203580) suppressed AGEs-induced iNOS expression and nitrite release from C6 glioma cells. AGEs activated p38 MAPK in C6 glioma cells, and this effect was blocked by genistein (20 microM), tyrphostin (30 microM), FPT inhibitor-II (20 microM), and SB203580 (10 microM). Taken together, our data suggest that AGEs may activate the pathways of tyrosine kinase and Ras to induce p38 MAPK activation, which in turn induces iNOS expression and NO production in C6 glioma cells.     

寡齿新银鱼同工酶及其与幼态持续的关系

采用聚丙烯酰胺垂直板状连续电泳方法,对幼态持续(neoteny)的寡齿新银鱼(Neosalanx oligodontis Chen)13种同工酶的18个位点进行研究,并进行乳酸脱氢酶热变性实验。结果表明ADH、GOT、α-GPDH、LDH、MDH、ME、POX和SOD表现出一定的幼态持续的特征,其中LDH由A、B两个位点编码,C位点不表达,是因为个体发育早期C基因尚未表达时发生幼态持续的结果。ES和IDH无幼态持续特征。α-AMY、FUM和CAT因数据不足,未进行分析。寡齿新银鱼同工酶基因表达中幼态特征的表现为研究鱼类幼态持续的产生机制和遗传基础提供线索。     

人三叶因子3在毕氏酵母中的表达及生物活性分析

利用PCR方法在人胎儿胎盘cDNA中扩增了人三叶因子3基因（hTFF3）,插入到含有AOX1启动子和α－因子信号肽序列的表达载体pPIC9K中,采用毕氏酵母表达系统对其进行了高效的分泌表达,并用G418筛选高拷贝整合转化子。2％甲醇诱导酵母表达48h后,上清经SDS－PAGE和Western印迹证明重组蛋白质以双体的形式分泌表达,占总蛋白质的45％,能被抗hTFF3抗体所识别。表达产物经S－Sepharose、Q-Sepharose离子交换及Sephacryl S-100纯化后纯度达到95％以上。体内生物学活性研究证实hTFF3可有效的防止盐酸诱导的大鼠胃溃疡。     

Correlates of Chlamydia and Gonorrhea Infection among Female Sex Workers: The Untold Story of Jiangsu,China

Objective(s)

To estimate the prevalence of sexually transmitted infections (STIs) among female sex workers (FSWs) in the Jiangsu Province, China and measure the association of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections with their potential correlates.

Design

A cross-sectional study on a representative sample of FSWs in Yangzhou and Changzhou cities of Jiangsu was conducted.

Methods

185 sex-work venues in Yangzhou and 174 in Changzhou were selected by stratified random sampling. 2972 FSWs (1108 in Yangzhou and 1864 in Changzhou), aged 15 years or more, who agreed to participate and provided blood sample for HIV and syphilis testing were interviewed in these venues. Cervical specimens from 849 randomly chosen participants were then tested for CT and NG.

Results

Proportions of young, school-educated, currently married FSWs who were living alone, migrated from other provinces and engaged in unprotected vaginal intercourse in past 3 months (UVI) were relatively high. Prevalence of HIV, syphilis, CT and NG were 0.20%, 4.88%, 14.61% and 5.42% respectively. Younger age, living alone or with persons other than partners/family members, engaging in UVI and having other STIs seemed to be associated with higher risk of CT or NG infection. Being divorced/widowed and working in middle/low-level venues were identified as additional risk factors for NG.

Conclusions

Based on a representative sample, this initial effort to identify the correlates of CT/NG infections among FSWs of Jiangsu revealed that focused interventions targeting high-risk FSWs are urgently required for controlling STI epidemics in Yangzhou and Changzhou where substantial number of STI cases were identified.     

Dynamic involvement of ATG5 in cellular stress responses

Autophagy maintains cell and tissue homeostasis through catabolic degradation. To better delineate the in vivo function for autophagy in adaptive responses to tissue injury, we examined the impact of compromised autophagy in mouse submandibular glands (SMGs) subjected to main excretory duct ligation. Blocking outflow from exocrine glands causes glandular atrophy by increased ductal pressure. Atg5^f/−;Aqp5-Cre mice with salivary acinar-specific knockout (KO) of autophagy essential gene Atg5 were generated. While duct ligation induced autophagy and the expression of inflammatory mediators, SMGs in Atg5^f/−;Aqp5-Cre mice, before ligation, already expressed higher levels of proinflammatory cytokine and Cdkn1a/p21 messages. Extended ligation period resulted in the caspase-3 activation and acinar cell death, which was delayed by Atg5 knockout. Moreover, expression of a set of senescence-associated secretory phenotype (SASP) factors was elevated in the post-ligated glands. Dysregulation of cell-cycle inhibitor CDKN1A/p21 and activation of senescence-associated β-galactosidase were detected in the stressed SMG duct cells. These senescence markers peaked at day 3 after ligation and partially resolved by day 7 in post-ligated SMGs of wild-type (WT) mice, but not in KO mice. The role of autophagy-related 5 (ATG5)-dependent autophagy in regulating the tempo, duration and magnitude of cellular stress responses in vivo was corroborated by in vitro studies using MEFs lacking ATG5 or autophagy-related 7 (ATG7) and autophagy inhibitors. Collectively, our results highlight the role of ATG5 in the dynamic regulation of ligation-induced cellular senescence and apoptosis, and suggest the involvement of autophagy resolution in salivary repair.Autophagy is a catabolic process that has an essential role in cellular adaptation to multiple types of stress by recycling of superfluous cellular material, safeguarding quality control in organelles, removing protein aggregates, and eliminating intracellular pathogens.¹ Conceptually, autophagy serves a pro-survival mechanism by providing sources of energy and biosynthetic building blocks during starvation, removing dysfunctional organelles and large aggregates toxic to cells to avoid unwarranted cell death. However, upon sustained stress conditions, cell death eventually takes place either by excessive autophagy or by the induction of apoptosis and/or necrosis pathways.² The ATG5, autophagy-related 5, has a pivotal role in autophagosome formation. Mouse neonates systemic deficient for ATG5 die within a day of birth,³ whereas mice depleted of Atg5 in selected tissues have abnormalities ranging from neurodegeneration⁴ and age-related cardiomyopathy⁵ to liver tumors.⁶Autophagy and senescence are two distinct, however functionally intertwined, cellular responses to stress.⁷ Cellular senescence is a state of stable growth arrest that is induced by telomere shortening, DNA-damage, oncogenes or other stresses. In general, senescence is a heterogeneous phenotype, which is characterized by a senescent-associated secretory phenotype (SASP), expression of senescence-associated β-galactosidase (SA-β-gal) and other senescent markers, and increased cell size.⁸ In culture system, inhibiting or enhancing autophagy leads to the opposite effect on premature senescence.^{9, 10, 11, 12} While premature senescence can be induced by a plethora of cell-extrinsic and cell-intrinsic stressors,¹³ little is known about the possible role of autophagy in modulating injury-induced cellular senescence in vivo. Rodent salivary duct ligation has been used as an experimental model system to study salivary gland atrophy, which often occurs in patients with Sjögren''s syndrome or receiving head and neck radiation therapy. Although autophagy induction has been implicated in the repair of rapamycin-treated, post-ligated salivary glands,^14,15 the roles played by autophagy in regulating the injury responses in submandibular glands (SMGs) have not been explored.To explore how autophagy contributes to salivary (patho)physiology, we established a transgenic mouse model deficient for ATG5 in the salivary acinar cells. Previously, we have identified a role for basal autophagy in salivary homeostatic mechanisms that restrict acinar cell size and the number of secretory granules.¹⁶ Here, we report that ligation of the major SMG excretory duct triggers the glandular atrophy and the induction of autophagy. By comparing the acute and subacute stress responses from autophagy-impaired and -competent SMGs with duct obstruction, we established the intrinsic roles of ATG5-dependent autophagy in modulating salivary inflammatory responses, stress-induced senescence and cell death, which all occur sequentially in response to tissue injury. Our results provide in vivo evidence that stress-induced autophagic response is indispensable for resolving premature senescence in duct cells of the ligated glands, whereas ATG5 deficiency leads to delayed acinar cell death.     

低温条件下不同抗寒性薰衣草内源激素的变化

为揭示薰衣草内源激素与抗寒能力的关系,以抗寒性相对较强的狭叶薰衣草和抗寒性较弱的宽叶薰衣草为试验材料,在田间自然条件下,于越冬前不同降温时期分别对叶片和根系取样,采用酶联免疫吸附法（ELISA）测定和分析了内源激素ABA、GA3、IAA和ZR的含量变化。结果表明,两种薰衣草的叶片和根系中的ABA、IAA和ZR的含量随气温的降低均表现为先升高后降低的趋势,但GA3表现为持续下降的趋势。抗寒性强的狭叶薰衣草叶片和根系中的ABA、IAA和ZR含量均高于抗寒性弱的宽叶薰衣草,而GA3小于宽叶薰衣草。对薰衣草越冬起重要作用的内源激素是ABA。     

Review of the Chinese species of the genus Varma Distant (Hemiptera,Fulgoromorpha, Tropiduchidae), with description of two new species

Two new species of Varma Distant, 1906, V. falcata Chang & Chen, sp. n. (China: Guizhou) and V. lobata Chang & Chen, sp. n. (China: Guizhou) are described and illustrated. The female genitalia of four speices including two known species are described and illustrated for the first time. The diagnostic characters of this genus are redefined. A checklist to the species of Varma in China is given. The Keys on male and female genitalia to the Chinese species of Varma are provided.