菊花总黄酮含药血清对干眼细胞模型AR、NF-κB磷酸化蛋白及TGF-β1表达的影响＊

目的 观察菊花总黄酮在体外条件下对泪腺上皮细胞中雄激素受体（Androgen Receptor，AR）的调控作用及对下游的核因子κB（Nuclear Factor kappa B，NF-κB）、转化生长因子-β1（transforming growth factor-beta1，TGF-β1）的表达影响。方法 以过氧化氢（H₂O₂）诱导泪腺上皮细胞凋亡，建立细胞模型，设立无雄激素培养空白对照组、含雄激素培养对照组、无雄激素培养黄酮治疗组。以MTT法摸索含药血清最佳干预加入量；免疫印迹实验（Western Blot）以及实时荧光定量PCR法（Quantitative Real-Time PCR，qPCR）检测泪腺上皮细胞中AR、NF-κB及TGF-β1的表达情况；并应用雄激素受体阻断剂氟他胺进行AR阻断后再次检测上述指标 。结果 MTT法检测后计算得出含药血清干预细胞的最终浓度为13.2%；无雄激素培养黄酮治疗组及含雄激素培养对照组中AR、NF-κB及TGF-β1的表达增强，与空白对照组对比有显著差异（P<0.01）；无雄激素培养黄酮治疗组中NF-κB表达量比含雄激素培养对照组表达量低，两者间的差异有统计学意义。氟他胺及含药血清干预后，无雄激素培养黄酮治疗组及含雄激素培养对照组中AR及NF-κB的表达未见提高，无统计学意义。结论 菊花总黄酮对泪腺上皮细胞可能存在拟雄激素效应，从而使AR、NF-κB表达增加，并通过上调TGF-β1的表达，可能起到在泪腺局部的抗炎作用。     

Sex differences in the regulation of body weight

Obesity and its associated health disorders and costs are increasing. Males and females differ in terms of how and where body fat is stored, the hormones they secrete in proportion to their fat, and the way their brains respond to signals that regulate body fat. Fat accumulation in the intra-abdominal adipose depot is associated with the risk for developing cardiovascular problems, type-2 diabetes mellitus, certain cancers and other disorders. Men and postmenopausal women accumulate more fat in the intra-abdominal depot than do pre-menopausal women, and therefore have a greater risk of developing metabolic complications associated with obesity. The goal of this review is to explore what we know about sexual dimorphisms in adipose tissue accrual and deposition. Elucidating the mechanisms by which sex hormones may modulate the way in which fat is accumulated and stored is a critical area of research due to the prevalence of obesity and the metabolic syndrome, and the rapid increase in propensity for these diseases following menopause.     

邯郸市家庭护理人才状况的调查分析

随着现代医学模式的改变,社区护理已成为护理专业发展的方向.家庭护理是以家庭为单位的整体护理模式[1],是社区护理的重要组成部分,家庭护理人才状况是影响社区护理质量的重要因素.2007年7月-2007年11月,对邯郸市家庭护理人才状况、培养和需求等问题进行调查分析.现介绍如下.     

牵张成骨中植入物的临床应用特点

目的:阐述在领骨的牵张成骨过程中植入物的临床应用进展,并评价其生物相容性.方法:检索者为第一作者,分别以"颌骨,牵张成骨,治疗"和"Jaw,distraction osteogenesis,treatment"为检索词,在中国期刊全文数据库(CNKI:1989/2009)和Medline database数据库(1989/2009),采用电子检索的方式进行文献检索,共检索到56篇文章,按纳入和排除标准对文献进行筛选,共纳入20篇文章.从牵张成骨的治疗进展进行总结,对牵张成骨植入物的临床应用进展进行探讨.结果:牵张成骨植入物分为口内牵张器和口外牵张器,可进行转移盘的牵引.牵张器的选择应根据患者颅面骨具体情况而定,并符合患者的需要.牵张成骨己成为国内外口腔腔面外科及正畸处理诸多复杂牙颌面畸形、颌骨缺损等的重要手段.其与各种全身治疗、局部治疗和物理治疗等的联合应用可更有效的发挥成骨作用.目前牵张成骨植入物主要由金属材料构成.金属植入物在预防细菌滋生,保证植入物的牢固及牵引效应的发挥方面有很大优势,价格普遍昂贵.其中镍钛要忆合金丝成本较一般成品牵引器低的多.其固定装置及合金丝紧贴骨面,可完全植入组织内,且具有抗感染,可完全关闭内外伤口等优点.结论:牵张成骨是临床治疗牙颌面发育不足排齐牙齿及整复颌骨缺损畸形的行之有效的新方法. 金属材料植入物如植入小型的镍钦记忆合金丝的生物相容性较好.其他类型植入物的生物相容性有待提高.     

后循环梗死的临床表现分析

目的 了解后循环梗死常见临床症状、体征及特征性表现.方法 回顾性分析2004-2006年在我院神经内科住院的急性后循环梗死且资料完整的216例资料.所有患者在入院时均接受头颅MRI检查,证实在后循环供血区存在责任病灶.总结患者的临床症状和体征,分析后循环临床表现的特征及与病灶部位的关系.结果 患者常见的症状为偏侧肢体无力(81.9%)、口齿含糊(46.3%)、头晕(33.8%)、偏侧麻木(31.0%);最常见的体征足偏侧运动障碍(81.9%)、中枢性面舌瘫(61.1%)、构音障碍(46.3%)、偏侧感觉障碍(31.0%)及共济失调(30.1%);交叉性瘫痪发生率低(2.8%),单独的眩晕极少(1.4%);延髓麻痹、意识障碍、视力障碍及记忆障碍等症状有助于定位;经典的脑干综合征具有定位意义.结论 后循环梗夕匕临床表现复杂多变,特征性临床表现和临床综合征有助于后循环梗死的诊断.     

Structural basis for p50RhoGAP BCH domain–mediated regulation of Rho inactivation

Spatiotemporal regulation of signaling cascades is crucial for various biological pathways, under the control of a range of scaffolding proteins. The BNIP-2 and Cdc42GAP Homology (BCH) domain is a highly conserved module that targets small GTPases and their regulators. Proteins bearing BCH domains are key for driving cell elongation, retraction, membrane protrusion, and other aspects of active morphogenesis during cell migration, myoblast differentiation, and neuritogenesis. We previously showed that the BCH domain of p50RhoGAP (ARHGAP1) sequesters RhoA from inactivation by its adjacent GAP domain; however, the underlying molecular mechanism for RhoA inactivation by p50RhoGAP remains unknown. Here, we report the crystal structure of the BCH domain of p50RhoGAP Schizosaccharomyces pombe and model the human p50RhoGAP BCH domain to understand its regulatory function using in vitro and cell line studies. We show that the BCH domain adopts an intertwined dimeric structure with asymmetric monomers and harbors a unique RhoA-binding loop and a lipid-binding pocket that anchors prenylated RhoA. Interestingly, the β5-strand of the BCH domain is involved in an intermolecular β-sheet, which is crucial for inhibition of the adjacent GAP domain. A destabilizing mutation in the β5-strand triggers the release of the GAP domain from autoinhibition. This renders p50RhoGAP active, thereby leading to RhoA inactivation and increased self-association of p50RhoGAP molecules via their BCH domains. Our results offer key insight into the concerted spatiotemporal regulation of Rho activity by BCH domain–containing proteins.

Small GTPases are molecular switches that cycle between an active GTP-bound state and an inactive GDP-bound state and are primarily involved in cytoskeletal reorganization during cell motility, morphogenesis, and cytokinesis (1, 2). These small GTPases are tightly controlled by activators and inactivators, such as guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), respectively (3, 4), which are multidomain proteins that are themselves regulated through their interactions with other proteins, lipids, secondary messengers, and/or by posttranslational modifications (5–7). Despite our understanding of the mechanisms of action of GTPases, GAPs, and GEFs, little is known about how they are further regulated by other cellular proteins in tightly controlled local environments.The BNIP-2 and Cdc42GAP Homology (BCH) domain has emerged as a highly conserved and versatile scaffold protein domain that targets small GTPases, their GEFs, and GAPs to carry out various cellular processes in a spatial, temporal, and kinetic manner (8–15). BCH domain–containing proteins are classified into a distinct functional subclass of the CRAL_TRIO/Sec14 superfamily, with ∼175 BCH domain–containing proteins (in which 14 of them are in human) present across a range of eukaryotic species (16). Some well-studied BCH domain–containing proteins include BNIP-2, BNIP-H (CAYTAXIN), BNIP-XL, BNIP-Sα, p50RhoGAP (ARHGAP1), and BPGAP1 (ARHGAP8), with evidence to show their involvement in cell elongation, retraction, membrane protrusion, and other aspects of active morphogenesis during cell migration, growth activation and suppression, myoblast differentiation, and neuritogenesis (17–21). Aside from interacting with small GTPases and their regulators, some of these proteins can also associate with other signaling proteins, such as fibroblast growth factor receptor tyrosine kinases, myogenic Cdo receptor, p38-MAP kinase, Mek2/MP1, and metabolic enzymes, such as glutaminase and ATP-citrate lyase (17–26). Despite the functional diversity and versatility of BCH domain–containing proteins, the structure of the BCH domain and its various modes of interaction remain unknown. The BCH domain resembles the Sec14 domain (from the CRAL-TRIO family) (16, 27, 28), a domain with lipid-binding characteristics, which may suggest that the BCH domain could have a similar binding strategy. However, to date, the binding and the role of lipids in BCH domain function remain inconclusive.Of the BCH domain–containing proteins, we have focused on the structure and function of p50RhoGAP. p50RhoGAP comprises an N-terminal BCH domain and a C-terminal GAP domain separated by a proline-rich region. We found that p50RhoGAP contains a noncanonical RhoA-binding motif in its BCH domain and is associated with GAP-mediated cell rounding (13). Further, we showed previously that deletion of the BCH domain dramatically enhanced the activity of the adjacent GAP domain (13); however, the full dynamics of this interaction is unclear. Previously, it has been reported that the BCH and other domains regulate GAP activity in an autoinhibited manner (18, 21, 29, 30) involving the interactions of both the BCH and GAP domains, albeit the mechanism remains to be investigated. It has also been shown that a lipid moiety on Rac1 (a Rho GTPase) is necessary for its inactivation by p50RhoGAP (29, 31), which may imply a role in lipid binding. An understanding of how the BCH domain coordinates with the GAP domain to affect the local activity of RhoA and other GTPases would offer a previously unknown insight into the multifaceted regulation of Rho GTPase inactivation.To understand the BCH domain–mediated regulation of p50RhoGAP and RhoA activities, we have determined the crystal structure of a homologous p50RhoGAP BCH domain from S. pombe for functional interrogation. We show that the BCH domain adopts an intertwined dimeric structure with asymmetric monomers and harbors a unique RhoA-interacting loop and a lipid-binding pocket. Our results show that the lipid-binding region of the BCH domain helps to anchor the prenylation tail of RhoA while the loop interacts directly with RhoA. Moreover, we show that a mutation in the β5-strand releases the autoinhibition of the GAP domain by the BCH domain. This renders the GAP domain active, leading to RhoA inactivation and the associated phenotypic effects in yeast and HeLa cells. The released BCH domain also contributes to enhanced p50RhoGAP–p50RhoGAP interaction. Our findings offer crucial insights into the regulation of Rho signaling by BCH domain–containing proteins.     

Designer broad-spectrum polyimidazolium antibiotics

For a myriad of different reasons most antimicrobial peptides (AMPs) have failed to reach clinical application. Different AMPs have different shortcomings including but not limited to toxicity issues, potency, limited spectrum of activity, or reduced activity in situ. We synthesized several cationic peptide mimics, main-chain cationic polyimidazoliums (PIMs), and discovered that, although select PIMs show little acute mammalian cell toxicity, they are potent broad-spectrum antibiotics with activity against even pan-antibiotic-resistant gram-positive and gram-negative bacteria, and mycobacteria. We selected PIM1, a particularly potent PIM, for mechanistic studies. Our experiments indicate PIM1 binds bacterial cell membranes by hydrophobic and electrostatic interactions, enters cells, and ultimately kills bacteria. Unlike cationic AMPs, such as colistin (CST), PIM1 does not permeabilize cell membranes. We show that a membrane electric potential is required for PIM1 activity. In laboratory evolution experiments with the gram-positive Staphylococcus aureus we obtained PIM1-resistant isolates most of which had menaquinone mutations, and we found that a site-directed menaquinone mutation also conferred PIM1 resistance. In similar experiments with the gram-negative pathogen Pseudomonas aeruginosa, PIM1-resistant mutants did not emerge. Although PIM1 was efficacious as a topical agent, intraperitoneal administration of PIM1 in mice showed some toxicity. We synthesized a PIM1 derivative, PIM1D, which is less hydrophobic than PIM1. PIM1D did not show evidence of toxicity but retained antibacterial activity and showed efficacy in murine sepsis infections. Our evidence indicates the PIMs have potential as candidates for development of new drugs for treatment of pan-resistant bacterial infections.

AMPs and AMP mimics have attracted considerable attention as candidates for therapeutic development (1). The basic design elements include a region of charged residues, generally cationic residues, enabling interaction with bacterial cell surfaces, combined with a hydrophobic nature in AMPs (2). Unfortunately, AMPs and related polymers, in general, have one or more issues that limit their use as broad-spectrum antibiotics. Some are quite toxic to human cells, the potency of some is not adequate for human administration, others are sensitive to salt at levels present in human fluids, and some are too difficult and expensive to synthesize (3, 4). One broad-spectrum antimicrobial peptide, CST has seen increased recent use as a last resort antibiotic. CST is believed to kill bacteria by virtue of its ability to disrupt membrane integrity (5). This antibiotic requires intravenous administration and is nephrotoxic (6). The emergence of CST-resistant pathogens has also become a significant problem (7). We are unaware of any new broad-spectrum AMPs that have advanced to clinical trials.Imidazolium (IM) salts are antimicrobials (8), and there is an emerging literature on antimicrobial activity of side-chain and main-chain polyimidazolium (PIM) salts with chemical structures that are in some ways similar to those we describe. Although PIMs are potent antimicrobials, there are biocompatibility problems hindering their development, and some have somewhat limited activity spectra. As with other AMPs, there have been toxicity issues, potency issues, and delivery issues as many have large molecular masses, and there is little known about mammalian cell toxicity or mechanism of action (9–12).Here we show that members of a series of PIMs we designed and synthesized are potent broad-spectrum antibacterial compounds. We selected two for further analysis and showed they retain activity even against pan-antibiotic-resistant bacteria. Unlike CST and many other AMPs, which disrupt bacterial membranes, our model PIM is bactericidal without disrupting bacterial membranes. Our experiments provide insights about mechanism of action, the potential for the emergence of PIM resistance, and indicate PIMs are effective against a model gram-negative and a model gram-positive pathogen in murine infection models.     

成都地区心血管病高尿酸血症及相关因素—7288例人群分析

背景 过去认为中国人高尿酸血症及痛风较少，近年临床发现高尿酸血症不少见，且有增多趋势，可能与生活方式改变有关。本研究目的是观察人群中当前高尿酸血症状况及其有关因素。对象与方法 采用整群抽样，对成都市区有代表性人群进行人口统计资料，社会经济状况，体重指数（BMI＝体重kg/身高m^2)血压及某些危险因素综合调查。本研究是危险因素系列研究中有关血尿酸及其有关因素部分，对象为18－80岁，7288例长年居住成都年以上人群。结果 本研究人群平均血尿酸水平为332．5μmol/L（男性373．9μmol/L明显高于女性286．6μmol/L，P＜0．01），以≥428μmol/L为高尿酸血症。本组高尿酸血症男性19．8％，女性5．1％，总计13．2％，男性为女性危险之3．9倍，血尿酸水平受高血压影响大，随血压增加而明显增加，不同血尿酸水平时伴随的危险因素亦不同，与多数危险因素（如血压，胆固醇，甘油三酯等）明显相关。在双变量与多变量回归分析中均证实上述观察结果。结论 高尿酸目前在中国（成都）人群并不少见，且有进一步增加趋势，血尿酸常与多种危险因素伴随，支持某些专家及研究提出的尿酸可能为重要心血管病危险因素的假说，应予重视。正常，合理及异常尿酸水平，尚待在研究得证据的基础上获得共识。     

The effect of glucosamine and/or chondroitin sulfate on the progression of knee osteoarthritis: a report from the glucosamine/chondroitin arthritis intervention trial

OBJECTIVE: Osteoarthritis (OA) of the knee causes significant morbidity and current medical treatment is limited to symptom relief, while therapies able to slow structural damage remain elusive. This study was undertaken to evaluate the effect of glucosamine and chondroitin sulfate (CS), alone or in combination, as well as celecoxib and placebo on progressive loss of joint space width (JSW) in patients with knee OA. METHODS: A 24-month, double-blind, placebo-controlled study, conducted at 9 sites in the United States as part of the Glucosamine/Chondroitin Arthritis Intervention Trial (GAIT), enrolled 572 patients with knee OA who satisfied radiographic criteria (Kellgren/Lawrence [K/L] grade 2 or grade 3 changes and JSW of at least 2 mm at baseline). Patients with primarily lateral compartment narrowing at any time point were excluded. Patients who had been randomized to 1 of the 5 groups in the GAIT continued to receive glucosamine 500 mg 3 times daily, CS 400 mg 3 times daily, the combination of glucosamine and CS, celecoxib 200 mg daily, or placebo over 24 months. The minimum medial tibiofemoral JSW was measured at baseline, 12 months, and 24 months. The primary outcome measure was the mean change in JSW from baseline. RESULTS: The mean JSW loss at 2 years in knees with OA in the placebo group, adjusted for design and clinical factors, was 0.166 mm. No statistically significant difference in mean JSW loss was observed in any treatment group compared with the placebo group. Treatment effects on K/L grade 2 knees, but not on K/L grade 3 knees, showed a trend toward improvement relative to the placebo group. The power of the study was diminished by the limited sample size, variance of JSW measurement, and a smaller than expected loss in JSW. CONCLUSION: At 2 years, no treatment achieved a predefined threshold of clinically important difference in JSW loss as compared with placebo. However, knees with K/L grade 2 radiographic OA appeared to have the greatest potential for modification by these treatments.