Xiao-Xu-Ming decoction extract regulates differentially expressed proteins in the hippocampus after chronic cerebral hypoperfusion

Xiao-Xu-Ming decoction has been widely used to treat stroke and sequelae of stroke. We have previously shown that the active fractions of Xiao-Xu-Ming decoction attenuate cerebral ischemic injury. However, the global protein profile and signaling conduction pathways regulated by Xiao-Xu-Ming decoction are still unclear. This study established a two-vessel occlusion rat model by bilateral common carotid artery occlusion. Rats were intragastrically administered 50 or 150 mg/kg Xiao-Xu-Ming decoction for 4 consecutive weeks. Learning and memory abilities were measured with Morris water maze. Motor ability was detected with prehensile test. Coordination ability was examined using the inclined screen test. Neuronal plasticity was observed by immunofluorescent staining. Differentially expressed proteins of rat hippocampus were analyzed by label-free quantitative proteomics. Real time-polymerase chain reaction and western blot assay were used to identify the changes in proteins. Results showed that Xiao-Xu-Ming decoction dramatically alleviated learning and memory deficits, and motor and coordination dysfunction, and increased the expression of microtubule-associated protein 2. Xiao-Xu-Ming decoction extract remarkably decreased 13 upregulated proteins and increased 39 downregulated proteins. The regulated proteins were mainly involved in oxidation reduction process, intracellular signaling cascade process, and protein catabolic process. The signaling pathways were mainly involved in ubiquitin mediated proteolysis and the phosphatidylinositol signaling system. Furthermore, there was an interaction among Rab2 a, Ptpn1, Ppm1 e, Cdk18, Gorasp2, Eps15, Capza2, Syngap1 and Mt-nd1. Protein analyses confirmed the changes in expression of MTND1. The current findings provide new insights into the molecular mechanisms of Xiao-Xu-Ming decoction extract's effects on chronic cerebral hypoperfusion.

Ramulus Cinnamomi extract attenuates neuroinflammatory responses via downregulating TLR4/MyD88 signaling pathway in BV2 cells

Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.

Benign giant-cell tumor of the common bile duct:A case report

Primary giant-cell tumors rarely arise in the common bile duct. We herein report a case of primary giant-cell tumor of the common bile duct. The patient was an 81-year-old male who was diagnosed with a well-defined 1.2-cm mass projecting into the lumen of the middle common bile duct. Excision of the gallbladder and extrahepatic bile duct and a Roux-en-Y cholangiojejunostomy were performed. Histologically, the tumor had no association with carcinomas of epithelial origin and was similar to giant-cell tumors of the bone. The tumor consisted of a mixture of mononuclear and multinucleated osteoclast-like giant cells. The mononuclear cells showed no atypical features, and their nuclei were similar to those of the multinucleated giant cells. CD68 was expressed on the mononuclear and multinucleated osteoclast-like giant cells, whereas CD163 immunoreactivity was restricted to the mononuclear cells. Six months after the operation, the patient was still alive and had no recurrence. The interest of this case lies in the rarity of this entity, the difficulty of preoperative diagnosis, and this tumor’s possible confusion with other malignant tumors.

Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia

Objective:To construct rapidly a full-length cDNA library from nanogram amounts total RIMA of Giardia lamblia(G.lamblia) trophozoites stocked in RNA stabilization reagent.Methods:Total RNA of Giardia was extracted using Trizol reagent.A full-length cDNA library of G.lamblia trophozoites was constructed by a long-distance PCR(LD-PCR) method.The recombinant rate and the coverage rate of full-length clones of the library were evaluated.The inserted fragments were identified and sequenced by PCR amplification.Results:The titer of cDNA library was 3.85×10~7 pfu/mL.The length of inserted fragments ranged from 0.4 to 2.5 kb,and the recombination efficiency accounted for 100%(20/20).The coverage rate of full-length clones is high(17/20). Conclusions:The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature.The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA.

Genetic analysis of walnut cultivars from southwest China:Implications for germplasm improvement

Walnuts are highly valued for their rich nutritional profile and wide medicinal applications.This demand has led to the intensification of breeding activities in major walnut production areas such as southwest China,in order to develop more superior cultivars.With the increasing number of cultivars,accurate identification becomes fundamental to selecting the right cultivar for grafting,industrial processing or development of new cultivars.To ensure proper identification of cultivars and understand the genetic structure of wild and cultivated material,we genotyped 362 cultivated and wild individuals of walnut trees from southwest China(with two additional populations from Xinjiang,plus three cultivars from Canada,France and Belgium) using 36 polymorphic microsatellite loci.We found relatively low indices of genetic diversity(H_(O)=0.570,H_(E)=0.404,N_(A)=2.345) as well as a high level of clonality(>85% of cultivars),indicating reliance on genetically narrow sources of parental material for breeding.Our STRUCTURE and PCoA analyses generally delineated the two species,though considerable levels of introgression were also evident.More significantly,we detected a distinct genetic group of cultivated Juglans sigillata,which mainly comprised individuals of the popular ’Yangbidapao’ landrace.Finally,a core set of 18 SSR loci was selected,which was capable of identifying 32 cultivars.In a nutshell,our results call for more utilization of genetically disparate material,including wild walnut trees,as parental sources to breed for more cultivars.The data reported herein will significantly contribute towards the genetic improvement and conservation of the walnut germplasm in southwest China.

New insights into the evolutionary history of Megacodon:Evidence from a newly discovered species

Megacodon is an ideal genus to study speciation and ecological adaptation in the Sino-Himalayan region.The genus contains two species distributed at different elevations and in two separate areas.However,studies of this genus have long been impeded by a lack of fieldwork on one of its species,Megacodon venosus.In this study,we collected specimens of two Megacodon species and found an extraordinary new species of Megacodon in Lushui county of north-west Yunnan province,which we have since named Megacodon lushuiensis.We propose new species based on both morphological and molecular evidence.The finding of this new species emphasized the importance of ecological divergence in the divergence of Megacodon stylophorus and its parapatric low-elevation Megacodon species.To identify genetic determinants that underlie adaptations to different elevations,we characterized transcriptomes of the new species M.lushuiensis,which is distributed at low elevations,and M.stylophorus,which is distributed at high elevations.Comparative transcriptome analysis identified 8926 orthogroups containing single-copy genes,and 370 orthogroups containing significantly positively selected genes.The set of positively selected genes was enriched into 25 Gene Ontology terms,including "response to water deprivation","response to osmotic stress",and "cellular response to external stimulus".Our results provide new insights into how ecological adaptation and speciation occurred in Megacodon and highlight the role of heterogeneous habitats in the speciation of plants in the Sino-Himalayan region.

Xiao-Xu-Ming decoction protects against chronic cerebral ischemia injury in rats and profile of differentially expression proteins analysis in hippocampus

OBJECTIVE Vascular dementia(VD) refers to a progressive decline in memory and cognitive function caused by chronic cerebral ischemia.2-Vessels occlusion(2-VO) has been widely used as a model of VD.Xiao-Xu-Ming decoction,a well-known traditional Chinese medicine prescrip.tion,has been widely used to treat stroke and sequelae of stroke.The present study was to investigate the mechanism of Xiao-Xu-Ming decoction(XXM) against chronic cerebral ischemia injury in rats.METHODS After XXM treatment,rats were performed a memory testing with Morris water maze and motor ability testing using prehensile test and inclined screen test.Neuronal plasticity was observed by immunofluorescent staining with MAP2 antibody.Differentially expressed proteins of rat hippocampus were analyzed by Label-free quantitative proteomics.RESULTS XXM significantly alleviated 2-VOinduced learning and memory deficits,motor ability dysfunction,and neuronal plasticity injury in rats.The mechanism might be involved in up-regulation of 39 proteins and down-regulation of 13 proteins in the hippocampus of rats after XXM treatment vs 2-VO group rats.Gene ontology and pathway analysis showed that the regulated proteins are mainly involved in oxidation reduction process,intracellular signaling cascade process,and protein catabolic process,etc.The signal pathways are mainly involved in ubiquitin mediated proteolysis and phosphatidylinositol signaling system.CONCLUSION Current findings provide new insights into the molecular mechanisms of XXM on chronic cerebral ischemia.

双跨转子的随机不确定性非参数建模与可靠性分析（英文）

目的:旋转机械由于工作环境复杂,在运行过程中会不可避免地受到各种不确定性因素的影响,从而引发转子系统的异常振动。因此,迫切需要对系统工作状态开展可靠性分析。本文将外部扰动不确定性与模型不确定性考虑在内,旨在建立转子系统运行状态的可靠性评估指标,丰富转子动力学理论体系,为工程应用提供参考。创新点:1.采用非参数法进行建模,能够将外部扰动不确定性与模型不确定性同时包含在内;2.在非参数建模基础上,结合响应谱分析法进行可靠性计算,可避免对系统先验知识的需求并降低计算过程的复杂性;3.将短周期预测理论扩展应用于可靠性分析验证。方法:1.借助非参数法建立转子系统的随机不确定性模型;2.结合响应谱分析法推导出系统可靠性指标计算式;3.采用短周期预测方法对模拟数据统计指标进行计算与验证。结论:1.本方法可用于评估大型复杂旋转机械系统的可靠性,尤其对于服役时间较长导致系统参数出现不确定性变化的情形;2.本研究结果可为大型复杂旋转机械的设计、运行和控制提供理论基础,同时也可以为其他类型机械设备的可靠性分析和预测方法提供参考。