胃镜下重酒石酸去甲肾上腺素喷洒与注射治疗急性 非静脉曲张性上消化道出血的疗效比较

目的探讨和比较胃镜下重酒石酸去甲肾上腺素喷洒与注射治疗急性非静脉曲张性上消化道出血(ANVUGIB)的临床疗效。方法选取2015年12月-2017年11月该院收治的92例ANVUGIB患者作为临床研究对象,采用随机数字表法,将入选患者随机分为观察组47例和对照组45例。观察组患者给予胃镜下重酒石酸去甲肾上腺素注射的临床药物治疗,对照组患者则给予胃镜下重酒石酸去甲肾上腺素喷洒的临床药物治疗,并分别对两组患者的临床治疗情况、临床指标变化情况和不良反应发生情况进行比较和分析。结果与对照组患者相比,观察组患者临床治疗的显效率57.45%(27/47)和总有效率91.49%(43/47)均明显提高,而无效率8.51%(4/47)则明显降低,差异均有统计学意义(P 0.05);观察组患者止血成功率91.49%(43/47)明显提高,差异有统计学意义(P 0.05),同时再出血率6.38%(3/47)和急诊手术率2.13%(1/47)则均有所降低,但差异均无统计学意义(P0.05);观察组患者腹部不适、大便频繁、胀气和血压不稳等不良反应总发生率10.65%(5/47)明显降低,差异有统计学意义(P 0.05)。结论胃镜下重酒石酸去甲肾上腺素注射治疗ANVUGIB较喷洒给药疗效更为确切,且预后效果好,并发症少。     

血管内皮生长因子在妊娠小鼠子宫GMG细胞中的表达

目的探讨颗粒子宫腺(GMG)细胞中血管内皮生长因子(VEGF)的表达。方法取妊娠13~15 d昆明种小鼠的子宫腺细胞区,经组织块培养提纯GMG细胞,应用免疫组织化学及Western blot法显示GMG细胞中VEGF。结果组织块培养时GMG细胞生长状况好,细胞纯度高,在GMG细胞及其培养液中均有VEGF表达。结论GMG细胞表达和分泌VEGF。     

基于知识图谱构建和定性访谈法探析张忠德教授辨治间质性肺病临床特征与方药规律＊

目的 构建个性化知识图谱技术和定性访谈法结合，进一步挖掘张忠德教授辨治间质性肺疾病临证特征与用药规律。方法 采用回顾性分析，系统收集张忠德教授广东省中医院门诊2010年8月至2020年8月治疗间质性肺疾病病历，按照诊断标准、纳入标准、排除标准，严格筛选后，通过广东省中医院大数据挖掘团队中医药大数据智能处理与知识服务系统进行数据挖掘分析，并通过多元化视觉定性访谈法，将定量与定性分析有效结合。结果 共筛选出347首方，共141味药物，常用药物频次 ≥ 84次的药物有10味，其中党参、麦芽、黄芪、紫菀、白术等为核心用药，通过症状与药物推理知识地图显示，党参、炒麦芽、黄芪、大枣、太子参、山萸肉、巴戟天等为主要治疗用药；临证遣方用药知识关联分析，得知咳嗽、耳鸣、心悸、水肿、头痛、胸闷、恶寒等为多见，针对咳嗽，首选紫苏子、橘红、桂枝等温肺降气通阳之品等；频繁聚集显示，常用药对炒麦芽-炒白术、黄芪-党参、黄精-菟丝子、前胡-紫菀、炙枇杷叶-浙贝母等；聚类分析结果得到4组关系密切的聚类新药物组合；以脾为中轴，肺肾共扶为主的“平调五脏论”，分期阶梯辨治间质性肺疾病。结论 张忠德教授认为间质性肺疾病，虚实夹杂为多见，应从肺、脾、肾着手，采用平调五脏论分期阶梯辨治，用药配伍精简，以和为贵，以平为期。     

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.     

Alpha‐phellandrene‐induced DNA damage and affect DNA repair protein expression in WEHI‐3 murine leukemia cells in vitro

Although there are few reports regarding α‐phellandrene (α‐PA), a natural compound from Schinus molle L. essential oil, there is no report to show that α‐PA induced DNA damage and affected DNA repair associated protein expression. Herein, we investigated the effects of α‐PA on DNA damage and repair associated protein expression in murine leukemia cells. Flow cytometric assay was used to measure the effects of α‐PA on total cell viability and the results indicated that α‐PA induced cell death. Comet assay and 4,6‐diamidino‐2‐phenylindole dihydrochloride staining were used for measuring DNA damage and condensation, respectively, and the results indicated that α‐PA induced DNA damage and condensation in a concentration‐dependent manner. DNA gel electrophoresis was used to examine the DNA damage and the results showed that α‐PA induced DNA damage in WEHI‐3 cells. Western blotting assay was used to measure the changes of DNA damage and repair associated protein expression and the results indicated that α‐PA increased p‐p53, p‐H2A.X, 14‐3‐3‐σ, and MDC1 protein expression but inhibited the protein of p53, MGMT, DNA‐PK, and BRCA‐1. © 2014 Wiley Periodicals, Inc. Environ Toxicol 30: 1322–1330, 2015.     

Association of mitochondrial deoxyribonucleic acid 16189 variant (T->C transition) with metabolic syndrome in Chinese adults

OBJECTIVE: A common variant in mitochondrial DNA (mtDNA) at bp 16189 (T-->C transition) has been associated with small birth size, adulthood hyperglycemia, and insulin resistance in Caucasians. In this study, we investigated whether mtDNA 16189 variant is associated with metabolic syndrome in Chinese subjects. METHODS: Six hundred fifteen Chinese adults, aged 40 yr or older, were recruited in this study. The 16189 variant of mtDNA was detected using PCR and restriction enzyme digestion. Metabolic syndrome was diagnosed on modified National Cholesterol Education Program Adult Treatment Panel III guidelines, using body mass index (BMI) instead of waist circumference. An association study was performed with chi2 test and logistic regression analysis. RESULTS: The prevalence of the 16189 variant was higher in patients with metabolic syndrome than in those without: 44% (125 of 284) vs. 33.2% (110 of 331) (P = 0.006). The association between this 16189 variant of mtDNA and metabolic syndrome (P = 0.021) remained significant even after correcting for age and BMI. As to the individual traits, the prevalence of fasting plasma glucose of at least 110 mg/dl (> or =6.1 mmol/liter) [(51.5% (121 of 235) vs. 42.1% (160 of 380); P = 0.023], type 2 diabetes mellitus [48.1% (113 of 235) vs. 39.2% (149 of 380); P = 0.031], and hypertriglyceridemia [44.3% (104 of 235) vs. 35.8% (136 of 380); P = 0.037] were significantly higher in subjects harboring the 16189 variant of mtDNA than those with the wild type. However, the prevalence of hypertension [53.2% (125 of 235) vs. 47.6% (181 of 380); P = 0.180], BMI greater than 25 kg/m2 [48.5% (114 of 235) vs. 43.9% (167 of 380); P = 0.270], and low high-density lipoprotein cholesterol [61.3% (144 of 235) vs. 54.7% (208 of 380); P = 0.111] did not reach a significant difference between the two groups. Furthermore, there was a trend of increasing frequency of occurrence of the 16189 variant in individuals having an increasing number of components of metabolic syndrome (Ptrend < 0.005). CONCLUSION: Our data strongly suggest that mtDNA 16189 variant underlies susceptibility to metabolic syndrome in the Chinese population.     

Sealing of anodized magnesium alloy AZ31 with MgAl layered double hydroxides layers

In this work, anodized magnesium alloy AZ31 with and without boiling water sealing was pre-prepared, and then MgAl-layered double hydroxide (LDH) films were fabricated on it through hydrothermal chemical conversion of the pre-prepared anodic layer. The morphology, structure, and composition of the films were characterized by XRD, SEM, EDS, FT-IR, XPS and GDOES. It was found that the porosity of the films was reduced after in situ fabrication of the LDHs. The effects of boiling water sealing treatment on the anodized substrate were also discussed. Moreover, the polarization curve, EIS, and immersion tests showed that LDHs fabricated on the anodized substrate with boiling water sealing treatment exhibited a significant long period of protection for the substrate.

In this work anodized magnesium alloy AZ31 with and without boiling water sealing was pre-prepared, and then MgAl-layered double hydroxide (LDH) films were fabricated on it through hydrothermal chemical conversion of the pre-prepared anodic layer.     

Optimal treatment allocation for placebo‐treatment comparisons in trials with discrete‐time survival endpoints

In many randomized controlled trials, treatment groups are of equal size, but this is not necessarily the best choice. This paper provides a methodology to calculate optimal treatment allocations for longitudinal trials when we wish to compare multiple treatment groups with a placebo group, and the comparisons may have unequal importance. The focus is on trials with a survival endpoint measured in discrete time. We assume the underlying survival process is Weibull and show that values for the parameters in the Weibull distribution have an impact on the optimal treatment allocation scheme in an interesting way. Additionally, we incorporate different cost considerations at the subject and measurement levels and determine the optimal number of time periods. We also show that when many events occur at the beginning of the trial, fewer time periods are more efficient. As an application, we revisit a risperidone maintenance treatment trial in schizophrenia and use our proposed methodology to redesign it and compare merits of our optimal design. Copyright © 2015 John Wiley & Sons, Ltd.