Enhanced autophagic clearance of amyloid-βvia histone deacetylase 6-mediated V-ATPase assembly and lysosomal acidification protects against Alzheimer's disease in vitro and in vivo

Recent studies have suggested that abnormal acidification of lysosomes induces autophagic accumulation of amyloid-βin neurons,which is a key step in senile plaque formation.Therefore,resto ring normal lysosomal function and rebalancing lysosomal acidification in neurons in the brain may be a new treatment strategy for Alzheimer's disease.Microtubule acetylation/deacetylation plays a central role in lysosomal acidification.Here,we show that inhibiting the classic microtubule deacetylase histone deacetylase 6 with an histone deacetylase 6 shRNA or thehistone deacetylase 6 inhibitor valproic acid promoted lysosomal reacidification by modulating V-ATPase assembly in Alzheimer's disease.Fu rthermore,we found that treatment with valproic acid markedly enhanced autophagy.promoted clearance of amyloid-βaggregates,and ameliorated cognitive deficits in a mouse model of Alzheimer's disease.Our findings demonstrate a previously unknown neuroprotective mechanism in Alzheimer's disease,in which histone deacetylase 6 inhibition by valproic acid increases V-ATPase assembly and lysosomal acidification.

Cryptococcus splenic abscess in primary biliary cholangitis: a case report

Chronic liver diseases,including primary biliary cholangitis(PBC),are recognized as immunosuppressive conditions due to the impaired functionality of hepatic sinusoidal Kupffer cells.[1]PBC is specifically characterized by cholangitis and progressive liver pathology,which may eventually lead to cirrhosis.The compromised liver function and reduced efficacy of Kupffer cells in patients with PBC weaken their cell-mediated immune responses,increasing their susceptibility to opportunistic pathogens such as Cryptococcus.[2]Cryptococcosis is an infectious disease caused by pathogenic encapsulated yeasts of the genus Cryptococcus,which is distributed worldwide and exhibits diverse clinical presentations.[3]Most case reports on cryptococcal infections are associated with HIV infection or other conditions that compromise the immune system.[4]Case reports related to primary biliary cholangitis are rare.Therefore,this case report underscores a rare cryptococcal infection in a patient with primary biliary cholangitis,emphasizing the need to consider opportunistic infections in those with compromised liver function and immune dysregulation.

Determinants of continuous usage intention of branded apps in omni-channel retail environment: comparison between experience-oriented and transaction-oriented apps

Branded applications(apps)have become core touchpoints for improving consumer shopping experiences in omni-channel retailing,and many firms have developed different types of branded apps to provide additional value.Moreover,continuous usage intention is the key to improving enterprises’gain efficiency and consumers’brand loyalty.This study aims to reveal how branded apps achieve continuance intention from the perspective of consumer perceptions by combining the technology acceptance model and investigating the impact of differences in channel features on usage behavior between the two types of branded apps.An experiment was designed comparing transaction-and experience-oriented branded apps.A structural equation modeling technique was employed to validate the model based on the survey data of respondents from the experimental groups.The results show that the supportive role of omni-channel has a unique experience mechanism that promotes continuous usage intention.However,there are two discrepant results regarding the effect of perceived complementarity on perceived usefulness in transaction-and experience-oriented branded apps.The mediating role of perceived usefulness between perceived consistency,complementarity,ease of use and consumer satisfaction was supported in the experience-oriented apps but rejected in the transaction-oriented apps.

Non-Thermal Biological Effect of Graphene Far-Infrared Ray on Saccharomyces cerevisiae Cells

Graphene materials can emit far-infrared ray, but the biological effects of graphene far-infrared ray have not been studied. Furthermore, the non-thermal biological effect of far-infrared ray on organism has not been systematically studied independently of the thermal effect. The purpose of this study was to investigate the non-thermal biological effect of graphene far-infrared ray (gFIR) on Saccharomyces cerevisiae cells. In this work, stringent control of the cultivation conditions was carried out to ensure the stability and constancy of the culture and its temperature. Flow cytometry was used to detect the non-thermal effect of gFIR irradiation on cell membrane permeability, mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) content. Compared with the control group, cell membrane permeability of the gFIR exposure cells decreased by 4.7%, MMP increased by 16% and intracellular ROS reduced by 10.7%. The results revealed the valuable features of the non-thermal biological effect of gFIR on Saccharomyces cerevisiae cells, and the further analysis demonstrated that graphene far-infrared materials should have great application value in disease prevention and health promotion.

IL-17及其相关细胞因子在腹型过敏性紫癜患儿外周血和肠黏膜中的变化

背景肠黏膜屏障的损伤、免疫系统紊乱可能是腹型过敏性紫癜(henoch-schonlein purpura, HSP)的重要发病机制之一,然而免疫因子与肠黏膜屏障损伤的关系尚不清楚.目的探讨外周血、肠黏膜白细胞介素-17(interleukin-17,IL-17)及其相关细胞因子的变化在腹型HSP发病机制中的作用,以及了解肠道菌群失调与腹型HSP发病机制的关系.方法选取26例急性期腹型HSP患儿为观察组, 16例健康体检儿童为健康对照组,收集两组儿童外周血、粪便标本;选取20例接受胃镜检查的急性期腹型HSP患儿为病例组,8例胃镜检查无明显黏膜病变的儿童为对照组,收集两组儿童十二指肠黏膜组织.采用流式细胞术检测外周血单个核细胞中辅助性T细胞-17(Th17)、细胞毒性T细胞-17(Tc17)细胞比例;ELISA检测血浆IL-17、IL-6、IL-23、γ-干扰素(IFN-γ)细胞因子含量;HE染色观察十二指肠黏膜活检组织病理改变;RT-PCR及免疫组化法检测十二指肠黏膜IL-17、IL-6、IL-23、IFN-γ mRNA及蛋白表达水平;细菌16S rDNA法检测肠道粪便中双歧杆菌、大肠杆菌的数量.结果观察组Th17与Tc17细胞比例均较健康对照组显著增高(P<0.05).观察组血浆IL-17、IL-6、IL-23水平均较健康对照组显著增高(P<0.05).病例组十二指肠黏膜组织病理改变主要为非特异性慢性炎症,部分可见血管炎性病变.病例组十二指肠黏膜IL-17、IL-6、IFN-γ mRNA及蛋白表达水平较对照组显著增高(P <0.05).观察组双歧杆菌数量、双歧杆菌/大肠杆菌比值较健康对照组显著下降(P<0.05).结论 IL-17及其相关细胞因子参与了腹型HSP患儿全身及肠黏膜局部的炎症反应,肠道菌群失调可能促进IL-17介导的炎症反应过程.与腹型HSP的发病及肠黏膜屏障损伤有关.

智能化无人机植保作业关键技术及研究进展

搭载高性能传感器和施药装备的农业植保无人机系统是精准农业领域具有代表性的智能装备之一。本研究首先从前端田间作业环境动态感知技术出发,阐述了无人机光谱成像遥感、多传感器融合的SLAM实时环境建模等技术在无人机植保作业方面的应用情况;然后对精准施药过程建模与优化控制有关的前沿技术进行了分析,包括旋翼下方风场结构演化及雾滴沉积过程仿真建模、多区域全覆盖条件下的智能作业路径规划、精准变量施药控制等;最后论述了作业效果评估与过程监管相关技术的发展现状,包括施药作业质量评价方法、基于云平台数据管理的全过程可视化监管等。在总结现有技术发展现状基础上,对未来智能化无人机植保关键技术发展趋势进行了预测,阐明了光谱图像获取与计算智能的深度学习识别聚类、基于高精度雾滴谱和风场模型预测的精准变量施药作业路径规划、基于传感器实时数据的作业质量评估和作业监管等新技术手段,将在遥感信息反演、药液飘移抑制、作业效率优化、施药过程管控等方面带来革命性的进步,使植保作业数据化、透明化,全过程可观可控,推动农业生产管理从机械化向智能化和智慧化迈进。

Microbubble-enhanced ultrasound exposure improves gene transfer in vascular endothelial cells

AIM： To explore the effects of ultrasound exposure combined with microbubble contrast agent （SonoVue） on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluorescent protein （pEGFP） transfer into human umbilical vein endothelial cells （HUVECs）. METHODS： HUVECs with fluorescein isothiocyanatedextran （FD500） and HUVECs with pEGFP were exposed to continuous wave （1.9 MHz, 80.0 mW/cm^2） for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytornetry, respectively. RESULTS： The percentage of FDS00-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone （24.0%± 5.5% vs 66.6% ± 4.1%, P 〈 0.001）. Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue （16.1% ± 1.9% vs 1.5% ± 0.2%, P 〈 0.001）. No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue （94.1% ± 2.3% vs 91.1% ± 4.1% ）. CONCLUSION： The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non- invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors.

Hypersonic boundary-layer transition on a flared cone

Transition on a flared cone with zero angle of at- tack was studied in our newly established Mach 6 quiet wind tunnel （M6QT） via wall pressure measurement and flow visualization. High-frequency pressure transducers were used to measure the second-mode waves＇ amplitudes and frequencies. Using pulsed schlieren diagnostic and Rayleigh scattering technique, we got a clear evolution of the second-mode disturbances. The second-mode waves exist for a long distance, which means that the second-mode waves grow linearly in a large region. Strong Mach waves are radiated from the edge of the boundary layer. With further development, the second-mode waves reach their maximum magnitude and harmonics of the second-mode instability appear. Then the disturbances grow nonlinearly. The second modes become weak and merge with each other. Finally, the nonlinear interaction of disturbance leads to a relatively quiet zone, which further breaks down, resulting in the transition of the bound- ary layer. Our results show that transition is determined by the second mode. The quiet zone before the final breakdown is observed in flow visualization for the first time. Eventual transition requires the presence of a quiet zone generated by nonlinear interactions.

Analysis on Factors Related to Rabies Epidemic in China from 2007-2011

To analyze features of the rabies epidemic in China between 2007 and 2011, identify factors influencing the epidemic and to provide a scientific basis for further control and prevention of rabies, Descriptive epidemiological methods and statistical analysis was used on data collected from the National Disease Reporting Information System between 2007 to 2011 and the National Active Surveillance System between 2007 and 2010. Our analysis shows that while the number of human rabies cases decreased year by year, the number of districts reporting cases did not show significant change. The situations in Guangdong, Guangxi, Guizhou and Hunan provinces clearly improved over the period but they remain provinces with high-incidence, and consequently influence the epidemic situation of surrounding provinces and possibly the whole country. Summer and autumn were high-incidence seasons. Farmers, students and pre-school children represent the high-risk populations, and rates of cases in farmers increased, those for students decreased, and pre-school children remained unchanged. Provinces with active surveillance programs reported a total of 2346 individual cases, of which 88.53% were associated with canines. Postexposure prophylaxis （PEP） of rabies cases was not significantly improved, whereas PEP in post-exposure population was good. In rural regions of China, canine density was reduced somewhat, and the immunization rate increased slightly. Finally we show that while the epidemic decreased 2007 to 2011 in China, cases continued to be diffused in certain regions. Lack of standardization of PEP on rabies eases was the main reason of morbidity. The high density and low immunization of dog in rural areas and the defective situation of PEP are still continuous occurrences in China and remain a cause for concern.

Establishment and Preliminary Application of a Rapid Fluorescent Focus Inhibition Test (RFFIT) for Rabies Virus

The World Health Organization (WHO) standard assay for determining levels of the rabies virus neutralization antibody (RVNA) is the rapid fluorescent focus inhibition test (RFFIT), which is used to evaluate the immunity effect after vaccination against rabies. For RFFIT, CVS-11 was used as the challenge virus, BSR cells as the adapted cells, and WHO rabies immunoglobulin (WHO STD) as the reference serum in this study. With reference to WHO and Pasteur RFFIT procedures, a micro-RFFIT procedure adapted to our laboratory was produced, and its specificity and reproducibility were tested. We tested levels of RVNA in human serum samples after immunization with different human rabies vaccines (domestic purified Vero cell rabies vaccine (PVRV) and imported purified chick embryo cell vaccine (PCECV)) using different regimens (Zagreb regimen and Essen regimen). We analyzed the levels of RVNA, and compared the immune efficacy of domestic PVRV and imported PCECV using different immunization regimens. The results showed that the immune efficacy of domestic PVRV using the Zagreb regimen was as good as that of the imported PCECV, but virus antibodies were generated more rapidly with the Zagreb regimen than with the Essen regimen. The RFFIT procedure established in our laboratory will enhance the comprehensive detection ability of institutions involved in rabies surveillance in China.

Expression of thymidine kinase mediated by a novel non-viral delivery system under the control of vascular endothelial growth factor receptor 2 promoter selectively kills human umbilical vein endothelial cells

AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.

Investigation and Analysis of Rabies Viral Infection and Distribution in China in 2005-2012

Dear Editor, We report the results of a preliminary investigation of data collected between 2005 and 2012. A National Human Rabies Surveillance program was initiated in high rabies incidence regions in 2005, and subsequently carried out nationally to monitor rabies situation. Our work presents a summary of epidemiological and etiologic rabies surveillance data collected since the implementation of the program.

Comparison of RFFIT Tests with Different Standard Sera and Testing Procedures

The World Health Organization （WHO） standard assay for determining antibody level is the rapid fluorescent focus inhibition test （RFFIT） and is used to determine the degree of immunity after vaccination against rabies. To compare the difference in RFFIT results between the laboratories of The National Institute of Infectious Disease in Japan （NIID） and the Chinese Centre for Disease Control （CCDC） as well the influence of the choice of standard serum （STD） for the detection, the two laboratories detection methods were simultaneously manipulated by RFFIT. The reference serums used in NIID and the WHO standard serum used in CCDC were compared in the same RFFIT detection to determine the titer of four sera samples C1, Sl, S2 and S4 in parallel, and the titers of the detected sera samples were calculated using the standard formula for neutralizing antibody titer. No significant difference was found in RFFIT methods from the two laboratories and the RFFIT testing procedures of the two laboratories have good consistency. However, different titers were obtained with the tentative internal standard serum （TI-STD） produced by adjusting to 2.0 IU of WHO standard serum in NIID and the WHO STD. The titer determined with the TI-STD was higher than that determined with WHO STD, This difference appears to be significant and requires further investigation

Molecular Characterization of China Human Rabies Vaccine Strains

To understand the molecular characteristics of China human rabies vaccine strains, we report the full-length genome of the aG strain and present a comprehensive analysis of this strain and almost all available lyssavirus genomes （58 strains） from GenBank （as of Jan 6, 2011）. It is generally considered that the G protein plays a predominant role in determining the pathogenicity of the virus, to this end we predicted the tertiary structure of the G protein of aG strain, CTN 181 strain and wild type strain HN 10 based on the crystal structure of Vesicular stomatitis virus （VSV） G. The predicted RABV G structure has a similar topology to VSV G and the ectodomain can be divided into 4 distinct domains DI - DIV. By mapping the characterized mutations to this structure between China vaccine strains and their close street strains, we speculate that the G303（P-H） mutations of CTN181 and HN10 causing D II 3D change may be associated with the attenuated virulence in both strains. Specifically, the two signature mutations （G165P and G231P） in the aG strain are withinβsheets, suggesting that both sites are of structural importance.

Analysis of transcriptome sequencing of sciatic nerves in Sprague-Dawley rats of different ages

An aging-induced decrease in Schwann cell viability can affect regeneration following peripheral nerve injury in mammals. It is therefore necessary to investigate possible age-related changes in gene expression that may affect the biological function of peripheral nerves. Ten 1-week-old and ten 12-month-old healthy male Sprague-Dawley rats were divided into young（1 week old） and adult（12 months old） groups according to their ages. mRNA expression in the sciatic nerve was compared between young and adult rats using next-generation sequencing（NGS） and bioinformatics（n = 4/group）. The 18 groups of differentially expressed mRNA（DEmRNAs） were also tested by quantitative reverse transcription polymerase chain reaction（n = 6/group）. Results revealed that（1） compared with young rats, adult rats had 3608 groups of DEmRNAs. Of these, 2684 were groups of upregulated genes, and 924 were groups of downregulated genes. Their functions mainly involved cell viability, proliferation, differentiation, regeneration, and myelination.（2） The gene with the most obvious increase of all DEmRNAs in adult rats was Thrsp（log2 FC = 9.01, P 〈 0.05）, and the gene with the most obvious reduction was Col2 a1（log2 FC = -8.89, P 〈 0.05）.（3） Gene Ontology analysis showed that DEmRNAs were mainly concentrated in oligosaccharide binding, nucleotide-binding oligomerization domain containing one signaling pathway, and peptide-transporting ATPase activity.（4） Analysis using the Kyoto Encyclopedia of Genes and Genomes showed that, with increased age, DEmRNAs were mainly enriched in steroid biosynthesis, Staphylococcus aureus infection, and graft-versus-host disease.（5） Spearman＇s correlation coefficient method for evaluating NGS accuracy showed that the NGS results and quantitative reverse transcription polymerase chain reaction results were positively correlated（rs = 0.74, P 〈 0.05）. These findings confirm a difference in sciatic nerve gene expression between adult and young rats, suggesting that, in peripheral nerves, cells and the microenvironment change with age, thus influencing the function and repair of peripheral nerves.

Investigation of the Evolutionary History of the Lyssaviruses

Dear Editor, We report the results of evolutionary history estimation of the lyssaviruses based on an analysis of the Glycoprotein (G) sequences gene using the BEAST software package. The most recent common ancestor (TMRCA) of all the lyssavirus strains was estimated to be approximately 5030 years (95% HPD 3988-6069 years), and there was a significant spread of the rabies virus throughout the world range in the last 200 years, consistent with significant time points in development and migration of human civilizations.

Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies

To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.

用焦虑-VAS评估音乐疗法对白内障手术老人焦虑的作用

目的:探讨术中音乐疗法对白内障手术老人焦虑的作用。方法:白内障手术老人患者40例,随机分为实验和对照组,每组各20例,实验组术中接受音乐疗法,对照组进行常规护理。采用焦虑-VAS分别于术前和术毕评估焦虑程度,同时记录血压、心率以及二者之积。结果:术前实验和对照组患者焦虑-VAS分别为7.2 &#177;2.0和7.3 &#177;1.9,差异无统计学意义(p >0.05)。实验组术毕焦虑-VAS为4.8 &#177;1.8,显著低于术前水平,差异有统计学意义(p 0.05);两组比较,实验组术毕焦虑-VAS显著低于对照组水平,差异有统计学意义(p 0.05),手术10分钟和术毕各参数显著增加,差异都有统计学意义(p <0.05),但实验组术中和术毕各参数增加程度小于对照组,差异无统计学意义(p <0.05)。结论:白内障手术中音乐疗法可缓解手术患者焦虑水平。

Effects of harvesting method and date on yield loss and seed quality of rapeseed

China is one of the major rapeseed production countries in the world,but harvesting mechanization was still backward,and high harvest loss was a key inhibiting factor for rapeseed production.To obtain optimum harvesting date for winter rape in the Yangtze River Valley of China,artificial simulated combine harvesting and artificial two-stage harvesting were adopted to find correlations between harvesting date,rape variety,seed oil content,protein content,grain moisture rate,rapeseed straw moisture content,rapeseed unthreshing rate,grain drop loss rate,thousand seeds weight(TSW),and harvesting economic coefficient(HEC).Analysis of variance showed that rapeseed oil content,protein content and TSW were correlated with rape variety;HEC was correlated with harvesting method and rape variety.Rape variety was the dominant factor of rapeseed oil content,protein content,TSW and HEC.Grain moisture rate,un-threshing rate,grain drop loss rate and straw moisture content were correlated with harvesting method and date.Harvesting date was the dominant factor of rapeseed moisture rate,un-threshing rate,grain drop loss rate and straw moisture rate.Single factor tests further proved that harvesting date had less impact on rapeseed oil content,protein content,TSW and HEC.It showed little correlation with rapeseed quality except grain moisture rate.The optimum harvesting date could be known by change in straw moisture content.The combined harvesting should be carried out during the grain moisture content of 15%-20%with TSW stabilized in the highest level.The twostage harvesting cutting should be carried out at the grain moisture content of 35%-40%.This research offered a reference to harvesting method and date for rape cultivated in the Yangtze River Valley.

黄单胞菌双组分信号转导系统的生物信息学分析

双组分信号转导系统是原核生物感知与响应刺激的重要代谢调节机制,广泛参与细菌的各种生理生化反应。本文采用生物信息学手段,利用多序列对比、系统进化树分析、跨膜区分析、二级结构预测等,对已完成全基因组测序的8株黄单胞菌中的双组分进行系统分类、结构分析和功能预测,初步构建了部分双组分信号转导系统的调控网络关系图,揭示了黄单胞菌致病性与双组份之间的联系,并初步阐明了黄单胞菌种间的进化关系,同时也为寻找新的药物靶标奠定了基础。