Evolutionary and phylogenetic analyses of 11 Cerasus species based on the complete chloroplast genome.

The subgenus Cerasus, one of the most important groups in the genus Prunus sensu lato, comprises over 100 species; however, the taxonomic classification and phylogenetic relationships of Cerasus remain controversial. Therefore, it is necessary to reconstruct the phylogenetic tree for known Cerasus species. Here, we report the chloroplast (cp) genome sequences of 11 Cerasus species to provide insight into evolution of the plastome. The cp genomes of the 11 Cerasus species (157,571-158,830 bp) displayed a typical quadripartite circular structure. The plastomes contain 115 unique genes, including 80 protein-coding genes, four ribosomal RNAs, and 31 transfer RNAs. Twenty genes were found to be duplicated in inverted repeats as well as at the boundary. The conserved non-coding sequences showed significant divergence compared with the coding regions. We found 12 genes and 14 intergenic regions with higher nucleotide diversity and more polymorphic sites, including matK, rps16, rbcL, rps16-trnQ, petN-psbM, and trnL-trnF. During cp plastome evolution, the codon profile has been strongly biased toward the use of A/T at the third base, and leucine and isoleucine codons appear the most frequently. We identified strong purifying selection on the rpoA, cemA, atpA, and petB genes; whereas ccsA, rps19, matK, rpoC2, ycf2 and ndhI showed a signature of possible positive selection during the course of Cerasus evolution. In addition, we further analyzed the phylogenetic relationships of these species with 57 other congenic related species.Through reconstructing the Cerasus phylogeny tree, we found that true cherry is similar to the flora of China forming a distinct group, from which P. mahaleb was separated as an independent subclade. Microcerasus was genetically closer to Amygdalus, Armeniaca, and Prunus (sensu stricto) than to members of true cherry, whereas P. japonica and P. tomentosa were most closely related to P. triloba and P. pedunculata. However, P. tianshanica formed a clade with P. cerasus, P. fruticosa, P. cerasus × P. canescens 'Gisela 6', and P. avium as a true cherry group. These results provide new insights into the plastome evolution of Cerasus, along with potential molecular markers and candidate DNA barcodes for further phylogenetic and phylogeographic analyses of Cerasus species.

Comparative transcriptome and metabolome analyses of cherry leaves spot disease caused by Alternaria alternata.

Alternaria alternata is a necrotrophic fungal pathogen with a broad host range that causes widespread and devastating disease in sweet cherry (Prunus avium). We selected a resistant cultivar (RC) and a susceptible cultivar (SC) of cherry and used a combined physiological, transcriptomic, and metabolomic approach to investigate the molecular mechanisms underlying the plant's resistance to A. alternata, of which little is known. We found that A. alternata infection stimulated the outbreak of reactive oxygen species (ROS) in cherry. The responses of the antioxidant enzymes and chitinase to disease were observed earlier in the RC than in the SC. Moreover, cell wall defense ability was stronger in the RC. Differential genes and metabolites involved in defense responses and secondary metabolism were primarily enriched in the biosynthesis of phenylpropanoids, tropane, piperidine and pyridine alkaloids, flavonoids, amino acids, and α-linolenic acid. Reprogramming the phenylpropanoid pathway and the α-linolenic acid metabolic pathway led to lignin accumulation and early induction of jasmonic acid signaling, respectively, in the RC, which consequently enhanced antifungal and ROS scavenging activity. The RC contained a high level of coumarin, and in vitro tests showed that coumarin significantly inhibited A. alternata growth and development and had antifungal effect on cherry leaves. In addition, differentially expressed genes encoding transcription factors from the MYB, NAC, WRKY, ERF, and bHLH families were highly expressed, they could be the key responsive factor in the response of cherry to infection by A. alternata. Overall, this study provides molecular clues and a multifaceted understanding of the specific response of cherry to A. alternata.

S100A9 activates NF-κB to promote TLR7 expression and inflammatory cytokine release in microglia / 神经解剖学杂志

Objective:To investigate the effect of S100 calcium-binding protein A9(S100A9)activation of nuclear factor kappa-B(NF-κB)on the upregulation of toll-like receptor 7(TLR7)expression and the release of inflammatory factors in microglia,as well as its underlying mechanism.Methods:The viability of BV2 microglia was assessed using CCK-8 kit.Transcriptome sequencing was employed to compare differential genes(DEGs)and identify target genes from the pool of differentially expressed genes.This analysis was complemented by GO analysis,KEGG enrichment analysis and the STRING database.The expression of TLR7 mRNA was verified by real time RT-PCR.The expressions of CD68 and CD206 were detected using immunofluorescence.The expressions of CD68,CD206,TLR7,p65,and p-p65 were detected using Western Blot.The level of interleukin 6(IL-6)and tumor necrosis factor alpha(TNF-α)were verified by ELISA.Results:Moderate concentrations of S100A9 had no inhibitory effect on microglial viability.Compared to the control group,the experimental group showed a significant increase in the expression level of CD68 pro-tein,while the CD206 protein was decreased.This suggests that S100A9 promotes the activation of BV2 microglia into pro-inflammatory types.TAK-242,an inhibitor of toll-like receptor 4(TLR4),significantly inhibited the expression levels of TNF-α and IL-6 after S100A9 stimulated BV2 cells.Activation of the TLR4/NF-κB pathway promoted the ex-pression of TLR7 protein.Conclusion:The moderate concentration of S100A9 can promote the polarization of microglia towards a proinflammatory direction.It also promotes the expression of TLR7 and the release of various inflammatory factors,including TNF-α and IL-6,through the activation of the TLR4/NF-κB pathway.This activation has an obvious proinflammatory effect.

Wavelet Element Modelling for Inviscid Fluid-Solid Coupling Problem based on Partitioned Approach.

To provide a simple numerical formulation based on fixed grids, a wavelet element method for fluid-solid modelling is introduced in this work. Compared with the classical wavelet finite element method, the presented method can potentially handle more complex shapes. Considering the differences between the solid and fluid regions, a damping-like interface based on wavelet elements is designed, in order to ensure consistency between the two parts. The inner regions are constructed with the same wavelet function in space. In the time and spatial domains, a partitioned approach based on Jacobi iteration is combined with the pseudo-parallel calculation method. Numerical convergence analyses show that the method can serve as an alternative choice for fluid-solid coupling modelling.

Tormentic acid confers protection against oxidative stress injury in rats with Parkinson's disease by targeting the Wnt/ß-catenin signaling pathway.

Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease (AD). Studies have shown that oxidative stress (OS) may contribute to the cascade of reactions leading to the degeneration of dopaminergic neurons in the brain. The present study investigated the protective effect of tormentic acid (TMA) on OS-induced injury in rat model of PD, and the underlying mechanism. Evaluation of learning and memorizing ability was done using Morris water maze (MWM) test. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx), and level of malondialdehyde (MDA) in substantia nigra were determined using enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expressions of ß-catenin, GSK-3ß, and GSK-3ß-Ser9 were determined using real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting. The effect of TA on cell viability and proliferation was determined in vitro on rat adrenal pheochromocytoma (PC12) cell line using MTT assay. The results showed that the escape latency of rats in negative control group was significantly higher than that in normal control group (p < 0.05). However, treatment with TMA significantly and time-dependently reduced the escape latency of the rats (p < 0.05). The extent of apoptosis was significantly  reduced after treatment with TMA (p < 0.05). Besides, treatment with TMA significantly increased the viability of brain cells (p < 0.05). The activities of SOD and GPx were significantly lower in negative control group than in normal control group, but were significantly increased after treatment with TMA (p < 0.05). The level of MDA was significantly higher in negative control group than in normal control group, but was significantly reduced after treatment with TMA (p < 0.05). The results of qRT-PCR and Western blotting showed that treatment with TMA significantly activated Wnt/ß-catenin signaling pathway (p < 0.05). TMA treatment significantly reversed the effect of 6-hydroxydopamine on the expression levels of these proteins and their mRNAs (p < 0.05). These results suggest that TMA confers protection against OS-induced injury in rats with PD by targeting the Wnt/ß-catenin signaling pathway.

Comparison of Diosgenin Content in Wild Dioscorea zingiberensis from Different Habitats / 中国药房

OBJECTIVE：To establish a method for the content determination of diosgenin in Dioscorea zingiberensis，and to compare the contents of diosgenin in wild D. zingiberensis from different habitats. METHODS：HPLC method was adopted. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of acetonitrile-water （50 ∶ 50，V/V） at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Established method was used to determine the contents of diosgenin in wild D. zingiberensis from different habitats （Shaanxi Shangluo，Shaanxi Ankang，Henan Neixiang，Yunnan Xuanwei，Sichuan Deyang，Hubei Shiyan，Hunan Zhangjiajie，Hunan Changde）. The differences of diosgenin content were compared. RESULTS：The linear range of diosgenin were 4.16-165.6 μg/mL （r＝     0.999 9）. RSDs of precision，reproducible and stability tests were all lower than 2％ （n＝5 or n＝6）. The average recovery of diosgenin was 101.18％ （RSD＝1.27％，n＝6）. The content of diosgenin in wild D. zingiberensis from different habitats were in descending order，i.e. Sichuan Deyang （1 405.36 μg/g）＞Shaanxi Shangluo （1 201.79 μg/g）＞Hunan Zhangjiajie （1 035.18 μg/g）＞Shaanxi Ankang （632.64 μg/g）＞Hunan Changde （598.64 μg/g）＞Yunnan Xuanwei （425.34 μg/g）＞Henan Neixiang （350.13         μg/g）＞Hubei Shiyan （338.39 μg/g）. CONCLUSIONS：Established method is simple，accurate and suitable for the content determination of diosgenin in D. zingiberensis. The contents of diosgenin in wild D. zingiberensis from different habitats are different significantly，and the content of diosgenin in the samples from Sichuan Deyang is the highest.

Study on the construction of medical statistics teaching model for postgraduates based on the structure of KAP / 中华医学教育探索杂志

Objective To construct the teaching model of medical statistics in universities of TCM through the questionnaire of knowledge,attitude and practice structure.Methods Medical statistics KAP questionnaire survey was conducted among postgraduates using proportional stratified sampling according to the grade.It describes the present situation and calculates the knowledge loss coefficient,knowledge demand index and behavioral application intensity of medical statistics.The Chi-square test was used to analyze the differences among different grades.Results 109 questionnaires were withdrawn.The recovery was 83.84％.45 cases (41.3％) were first year postgraduate,33 cases (30.3％) were 2nd year postgraduate,and 31 cases (28.4％) were 3rd year postgraduate.At the knowledge level,the knowledge loss coefficient of the second year postgraduates was 0.635,and there was no statistical difference (P=-0.651) vs the first year postgraduate (0.638).At the attitude level,Knowledge demand index had no statistical significance difference (P=0.213)between postgraduates in grade two (96.87％) and grade three (75.00％).Most medical postgraduates held positive attitudes toward medical statistics,hoping to strengthen the training of their application ability such as medical statistics in design and research thinking (29.18％),the statistical methods selection and the interpretation of the results (31.53％) and the statistical software operation (36.07％).At the practice level,the percentage of application intensity with statistical methods of third-year postgraduates (100.00％) was higher than the second-year postgraduates (37.50％),and the second-year postgraduates higher than the first-year postgraduates (20.45％).And with the growth of year,the percentage of application intensity was gradually increasing (P=O.O03).Among them,published scientific papers accounted for 16.5％,and 58.6％ paper grade concentrated in the core journals.Conclusion Application development should be enhanced by modularization teaching with practical issues.

Clinical and Pathologic Features of Multifocal and Multicentric Breast Cancer in Chinese Women: A Retrospective Cohort Study / 한국유방암학회지

PURPOSE: This study aims to analyze the clinical-pathological characteristics of multifocal and multicentric breast cancer (MMBC) in Chinese women. METHODS: Sixty-seven cases with MMBC were randomly collected and reviewed at seven hospitals in representative districts of China during 1999 to 2008. RESULTS: The incidence of MMBC in breast cancer in China was 1.75%. Compared to those with unifocal breast cancer, women with MMBC were more likely to have larger tumor size, lymph node metastasis (59.70% vs. 45.62%) and stage III to IV (46.26% vs. 21.10%). The peak age at onset of MMBC was 40 to 49 years old and has been gradually increasing during 1999 to 2008. Most of the MMBC women were treated with surgery and adjuvant therapy. CONCLUSION: In China, the incidence of MMBC in breast cancer is significantly lower than that in Western countries. Compared to unifocal breast cancer, MMBC is biologically more aggressive. Most MMBC women underwent mastectomy, instead of breast conservation surgery.

The molecular mechanisms of As_2O_3- induced un-terminal differentiation of HL-60 cells / 国外医学(肿瘤学分册)

To approach the effect of CCAAT/enhancer binding proteins (C/EBPs) on un-terminal differentiation of HL-60 cells after treatment with Arsenic Trioxide ( As_2O_3) . Methods The changes of cell morphology were observed by Wright staining, the alteration in the cell proliferation was determined by WST1 experiment and the NBT reduction assay was used to detect the differentiation condition of cells, determination and analysis cell cycle. The expressions of C/EBP? and C/EBP? mRNA in HL-60 cells exposed to ATRA and As_2O_3 were assayed by semi-quantitative RT-PCR. Results It was found that ATRA could up- regulate the mRNA expression of C/EBP? obviously, but down-regulate the mRNA expression of C/EBP?. As_2O_3 could up-regulate the mRNA expression of C/EBP? lightly, down-regulate the expression of C/EBP?. Conclusion Both of ATRA and As_2O_3 can down-regulate the mRNA expression of C/EBP?,but there is no significant difference between these two groups,ATRA and As2O3 can up- regulate the mRNA expression of C/EBP?, with significant differences (P

The exploration and practice on the culture of innovative manpower / 中华医学教育探索杂志

Graduate students are an important force for knowledge innovation.In order to explore the cultivation of innovative talents,our university reform curriculum courses,strengthen the graduate student instructing staff,improve the quality of scientific research thesis,enhance clinical skills training and build a base for innovation and post-graduate education and so on.