In Vitro Screening for Compounds Derived from Traditional Chinese Medicines with Antiviral Activities Against Porcine Reproductive and Respiratory Syndrome Virus

( Jia Cheng ),( Na Sun ),( Xin Zhao ),( Li Niu ),( Mei Qin Song ),( Yao Gui Sun ),( Jun Bing Jiang ),( Jian Hua Guo2 ),( Yuan Sheng Bai ),( Jun Ping He ),( Hong Quan Li ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.8

Seventeen compounds derived from traditional Chinese medicines (TCMs) were tested for their antiviral activity against porcine reproductive and respiratory syndrome virus (PRRSV) in vitro. Visualization with the cytopathologic effect (CPE) assay and the 3-(4, 5-dimethyithiazol- 2-yl)-2,5-diphenyltetrazolium bromide test were used to determine the 50% cytotoxic concentration (CC50) and 50% effective concentration (EC50) in cultured Marc-145 cells. Among the tested compounds, chlorogenic acid and scutellarin showed potential anti-PRRSV activity. The EC50 values were 270.8 ± 14.6 μg/ml and 28.21 ± 26.0 μg/ml and the selectivity indexes were >5.54 and 35.5, respectively. The time-of-addition and virucidal assay indicated that the anti-PRRSV activity of the two compounds could be due to their inhibiting the early stage of virus replication and/or inactivating the virus directly. The inhibition of the virus attachment was not observed in the adsorption inhibition assay. The inhibition ratios of chlorogenic acid and scutellarin were, respectively, 90.8% and 61.1% at the maximum non-cytotoxic concentrations. The results have provided a basis for further exploration of their antiviral properties and mechanisms in vivo. We believe that the chlorogenic acid and scutellarin have a great potential to be developed as new anti-PRRSV drugs for clinical application.

A Continuous Abnormal Speech Detection Method Based on Time Domain features Weighted

He Jun,Ji-chen Yang,Qing-hua Zhang,Guo-xi Sun,Jian-bing Xiong 보안공학연구지원센터 2014 International Journal of Control and Automation Vol.7 No.12

In this brief, a novel pathological continuous speech detection method based on time domain features weighted. First, different optimal threshold for time domain features, including zero crossing ratio, short-time energy and autocorrelation, are obtained from training speech data. Second, a difference evaluation technique is proposed, and with it, the difference of the same time domain feature selected from testing speech data and training speech data were obtained. Finally, to distinguish a given speech well, a novel weighting method based on difference evaluation for each kinds of time domain is employed, respectively. Experiments were conducted on the pathological speech database to prove the power and effectiveness of the proposed method. Results obtained shown that this method outperforms other early proposed time domain feature method, creating a more reliable technique for pathological continuous speech detection.

Effects of Wushu-Sanda Player`s Different Exercise Intensity on the Fatigue Substance, Oxidative Stress and Antioxidant Enzyme

( Guo Jian He ),( Zhi Hong Bao ),( Tian Yu Liu ),( Shuai Li ),( Jeong Ryong Chae ) 한국체육학회 2016 국제스포츠과학 학술대회 Vol.2016 No.1

Purpose: The purpose of this study was designed to examine the activation of Fatigue Substance, Oxidative Stress and Antioxidant Enzyme in Wushu Sanda players according to the different exercise intensity. Methods & Materials: Subjects of this four-week-long Wushu exercise program were divided into 2 groups: high intensity group (n=8), and low intensity group (n=8). Subjects participated in this program once a day (high intensity group:60mins, low intensity group:90mins) and 5 times per week (Monday to Friday). Every subjects’ fatigue substance (LAC, LDH), oxidative stress (MDA) and antioxidant enzyme (GPx, SOD, CAT) were measured before the program, immediately after exercise, after 30mins recovery and after 60mins recovery. All these parameters were measured again four-week latter. Statistics: ANOVA, paired t-test and repeated ANOVA were carried out to analyze the changes before and after the exercises, changes in terms of fatigue substance, oxidative stress and antioxidant enzyme, respectively. Statistical significance is established at the .05 level (p<.05). Result: 1. Change of LAC and LDH Pre and Post exercises, analysis of the LAC have statistically significant changes in recovery 30 and 60minutes than immediately after within each group (p<.05). But according to the time of measurement there are no statistically significant difference (p>.05). And, after four weeks, LAC in both groups are lower than the concentration of before exercise. Especially, the low intensity exercise group are lower than the high intensity exercise group. But all variables are no statistically significant differences(p<.05). Pre and Post exercises, analysis of the LDH was statistically significant changes in immediately after and recovery 60minutes within each group(p<.05). And according to the time of measurement were only significantly difference in the rest (p<.05). What’s more, the high intensity exercise group are lower than the low intensity exercise group (p>.05). 2. Change of MDA MDA in low intensity group was significantly higher after exercise and 30mins recovery than before exercise (p<.05). In high intensity group, no significant difference was found. 3. Change of GPx, SOD, CAT After the four weeks exercise, GPx in high intensity group shows significant differences after 30mins and 60mins recovery compared with the value of pre-exercise (p<.05). No significant difference was found in low intensity group. Compared to the value of SOD pre-exercise, the SOD of post-exercise shows significant differences after 30mins and 60mins recovery in high intensity group ( p<.05) while only immediately after exercise in low intensity group (p<.01 ). In CAT, a significant difference was found in high intensity group after the four weeks exercise (p<.05). No significant difference was found in low intensity group. Conclusion: After four weeks exercise, some differences were found in fatigue substance(LAC, LDH), oxidative stress(MDA) and antioxidant enzyme(GPx, SOD, CAT) in both groups: LDH was found to be increased in high intensity group after the four-week exercise; MDA was increased in low intensity group after exercise and 30mins recovery than before exercise; compared with the value of pre-exercise, GPx and SOD in high intensity group shows significant differences after 30mins and 60mins recovery; while in low intensity group, the SOD difference only occurred immediately after exercise; In CAT, a significant difference was found in high intensity group but no significant difference was found in low intensity group.

Prognostic Significance of Overexpression of EZH2 and H3k27me3 Proteins in Gastric Cancer

He, Long-Jun,Cai, Mu-Yan,Xu, Guo-Liang,Li, Jian-Jun,Weng, Zi-Jin,Xu, Da-Zhi,Luo, Guang-Yu,Zhu, Sen-Lin,Xie, Dan Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.7

The enhancer of zeste homolog 2 (EZH2) methyl transferase and histone 3 lysine 27 (H3K27me3) protein can repress gene transcription, and their aberrant expression has been observed in various human cancers. This study determined their expression levels in gastric cancer tissues with reference to clinicopathological features and patient survival. We collected 117 gastric cancer and corresponding normal tissues for immunohistochemistry analysis. In gastric cancers, 82/117 (70.1%) were positive for EZH2 and 66/117 (56.4%) for H3K27me3 proteins in contrast to only 5.41% and 7.25% of normal gastric mucosa specimens, respectively. Kaplan-Meier survival data showed the average overall and disease-free survival of EZH2 high expression patients was 25.2 and 20.2 months, respectively, shorter than that with EZH2 low expression (40.5 and 35.9 months). The average overall survival and disease-free survival of high H3K27me3 expression patients was 23.4 and 17.4 months, shorter than without H3K27me3 expression (37.6 and 34.5 months). The average overall survival and disease-free survival of patients with both EZH2 and H3K27me3 expression was 18.8 and 12.9 months, respectively, shorter than that with either alone (34.7 and 31.2 months) or with low levels of both (43.9 and 39.9 months). Multivariate Cox regression analysis showed that H3K27me3 and EZH2 expression, tumor size differentiation and clinical stage were all independent prognostic factors for predicting patient survival. This study demonstrated that detection of both EZH2 and H3K27me3 proteins can predict poor survival of gastric cancer patients, superior to single protein detection. In addition, H3K27me3 and EZH2 protein expression could predict lymph node metastasis.

Single Nucleotide Polymorphisms of the GnRHR Gene Associated with Reproductive Traits of Japanese Flounder (Paralichthys olivaceus)

He, Feng,Wen, Hai-Shen,Li, Ji-Fang,Yu, Da-Hui,Ma, Rui-Qin,Shi, Dan,Mu, Wei-Jie,Zhang, Yuan-Qing,Hu, Jian,Liu, Miao,Han, Wei-Guo,Zhang, Jia-Nan,Wang, Qing-Qing,Yuan, Yu-Ren,Liu, Qun Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.4

Gonadotropin-releasing hormone receptor (GnRHR) gene is expressed at the anterior pituitary gland and plays a key role in gonad development. This study aimed to investigate molecular genetic characteristics of the GnRHR gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of GnRHR gene on sex steroid level in Japanese flounder (Paralichthys olivaceus). We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the GnRHR gene in 75 individuals. We identified three SNPs in the GnRHR gene: P1 locus (C759A and C830T) in the coding region of exon2 which were both linked together and P2 locus (G984T) in the coding region of exon3, which added a new transcript factor (ADR1) and a new methylation site (CG). Only C830T of P1 leads to amino acid changes Thr266Ile. Statistical analysis showed that P1 was significantly associated with $17{\beta}$-estradiol ($E_2$) level (p<0.01) and gonadosomatic index (GSI) (p<0.05). Individuals with genotype BB of P1 had significantly higher serum $E_2$ levels (p<0.01) and GSI (p<0.05) than those of genotype AA or AB. Another SNP, P2, synonymous mutation, was significantly associated with GSI (p<0.05). Individuals with genotype AB of P2 had significantly higher GSI (p<0.05) than that of genotype AA. In addition, there was a significant association between one diplotype based on three SNPs and reproductive traits. The genetic effects for both serum $E_2$ level and GSI of diplotype D4 were super diplotypes (p<0.05). These results suggest that the SNPs in Japanese Flounder GnRHR are associated with $E_2$ level and GSI.

Characterization of Ha29, a Specific Gene for Helicoverpa armigera Single-nucleocapsid Nucleopolyhedrovirus

Guo, Zhong-Jian,An, Shi-Heng,Wang, Dun,Liu, Yan-He,Kumar, V. Shyam,Zhang, Chuan-Xi Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.3

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

Rs7574865 polymorphism of STAT4 and risk of anti-tuberculosis drug-induced hepatotoxicity in Chinese Han

Guo Chen,Wei Mao,Shou‑Quan Wu,Yu Wang,Gui‑Yi Ji,Miao‑Miao Zhang,Qian‑Qian Liu,Jian‑Qing He 한국유전학회 2017 Genes & Genomics Vol.39 No.11

Anti-tuberculosis drug-induced hepatotoxicity (ATDH) is the most frequent, and potentially fatal adverse effect in the treatment of tuberculosis (TB). The rs7574865 polymorphism in the signal transducer and activator of transcription 4 gene (STAT4) was reported to be associated with drug-induced liver injury. However, there was no study aimed to this association in Chinese patients. The aim of this study was to investigate the influence of STAT4 polymorphisms on the susceptibility to ATDH in a Chinese Han population. A total of 280 TB patients with the prescription of anti-TB therapy, of Chinese Han origin, were enrolled. They were followed up for 3 months and demographic, clinical, laboratory and therapeutic data at each visit were collected. Two single nucleotide polymorphisms (SNPs) (rs7574865 and rs7582694) of STAT4 were genotyped with the MassARRAY platform. The associations between SNPs and ATDH risk were analyzed by logistic regression adjusting for confounding factors. A total of 24 patients were diagnosed with ATDH and considered as the case group, and 33 patients were lost to follow-up; the remaining 223 subjects without ATDH were considered as the control group. There was strong linkage disequilibrium (LD) between rs7574865 and rs7582694 ( r2 = 0.928 and D′ = 1). No significant association was found between SNPs or haplotypes of STAT4 and ATDH after correction for confounding factors. This prospective study is the first to investigate the association of genetic polymorphisms of STAT4 and ATDH in Chinese individuals. There was no significant association between the rs7574865 of STAT4 and ATDH in a Chinese Han population.

Effects of the Maceration Enzymes on Evolution of Pyranoanthocyanins and Cinnamic Acids During the Cabernet Gernischet (Vitis vinifera L. cv.) Red Wine Making

Jian-Jun He,Fu-Liang Han,Qing-Quan Yu,Qiu-Hong Pan,Chang-Qing Duan,Guo-Li Cheng 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.3

The effects of the maceration enzymes on the evolution of 4-vinylphenols pyranoanthocyanins and cinnamic acids were investigated during the Cabernet Gernischet red wine making. The results showed that the contents of p-coumaric acid and caffeic acid in the maceration enzyme-treated wine (the treated wine) were higher than those in the maceration enzyme-untreated wine (the control wine) at the alcoholic fermentation (AF),maceration (M), and malolactic fermention (MLF) stages. Malvidin-3-O-glucoside-4-vinylphenol (Mv-glu-vp) and malvidin-3-O-(6-O-acetyl)-glucoside-4-vinylphenol (Mvacet-glu-vp) were formed at the AF and M stages in the treated wine, while they were formed at the end of MLF stage in the control wine. However, the contents of these 2kinds of 4-vinylphenol pyranoanthocyanins in the control wine were higher than those in the treated wine during aging. These results showed that the use of maceration enzymes in winemaking can promote p-coumaric acid and p-caffeic acid transformed to vinylphenols and accelerate the formation of pyranoanthocyanins.

Production of 3-hydroxypropionic acid by recombinant Klebsiella pneumoniae based on aeration and ORP controlled strategy

Jian-Guo Zhu,Xiao-Jun Ji,Jun Du,Shuang Li,Yue-Yue Ding,He Huang 한국화학공학회 2009 Korean Journal of Chemical Engineering Vol.26 No.6

A biosynthetic pathway for the production of 3-hydroxypropionic acid (3-HP) from glycerol was established in recombinant Klebsiella pneumoniae by introducing the aldehyde dehydrogenase gene from Escherichia coli. The activity of aldehyde dehydrogenase, which oxidized 3-hydroxypropionaldehyde (3-HPA) to 3-HP, was detected and 3-HP was produced by the recombinant strains. Three different oxygen supply strategies, associated with measuring the oxidoreduction potential (ORP) during the fermentation under these conditions, were adopted for higher production of 3-HP by the recombinant cells. About 0.8 g/l 3-HP and more 1,3-propanediol production by the recombinant Klebsiella pneumoniae were obtained under completely aerobic conditions. Under micro-aerobic conditions, 3-HP production could be increased to 2.2 g/l and 1,3-propanediol production was almost the same as in the original strain. Under the anaerobic conditions, 1,3-propanediol was the main product and about 1.3 g/l 3-HP was produced. Finally, 3-HP production of the recombinant strain was increased to 2.8 g/l under micro-aerobic condition with a further two-stage ORP controlled strategy.

Detection of the expression of a Bombyx mori Atypical Protein Kinase C in BmPLV-Infected Larval Midgut

( Jian Cao ),( Yuan Qing He ),( Guo Hui Li ),( Ke Ping Chen ),( Jie Kong ),( Feng Hua Wang ),( Jing Shi ),( Qin Yao ) 한국잠사학회 2011 International Journal of Industrial Entomology Vol.22 No.2

Protein kinase C (PKC) is involved in many cellular signaling pathways, it participates in many physiological processes, such as cell cycle, growth, proliferation, differentiation and apoptosis. To investigate the effect of PKC on the silkworm midgut tissue infection of Bombyx mori parvo-like virus (BmPLV), a B. mori atypical protein kinase C (BmaPKC) gene was cloned from larval midgut tissue, expressed in E. coli and purified. Additionally, the BmPLV susceptible silkworm strain and resistant silkworm strain were used to test the effect of the B. mori infection on BmPLV. The result showed that BmaPKC encodes a predicted 586 amino acid protein, which contains a C-terminal kinase domain and an N-terminal regulatory domain. The maximum expression amount of the soluble (His)6-tagged fusion protein was detected after 0.8 mmol/L IPTG was added and cultured at 21˚C. The (His) 6-tagged fusion protein revealed about 73 kDa molecular weight which confirmed by western blot and mass spectrography. Furthermore BmaPKC protein were detected at 0-72 h post-infection in BmPLVinfected larval midgut tissue, western blot showed that as time went on, the expression of BmaPKC increased gradually in susceptible strain, the expression quantity on 72 h is 5 times of 0 h. However, in resistant strain, the expression quantity is slightly lower than susceptible strain. But no significant change in resistant strain was observed as time went on. The available data suggest that BmaPKC may involve in the regulation of BmPLV proliferation.