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.

In Vitro Biological Characterization of DCUN1D5 in DNA Damage Response

Guo, Wei,Li, Guo-Jun,Xu, Hong-Bo,Xie, Jie-Shi,Shi, Tai-Ping,Zhang, Sheng-Zhong,Chen, Xiao-Hong,Huang, Zhi-Gang Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Background: Novel prognostic biomarkers or therapeutic molecular targets for laryngeal squamous cell carcinoma (LSCC) are an urgent priority. We here sought to identify multiple novel LSCC-associated genes. Methods: Using high-density microarray expression profiling, we identified multiple genes that were significantly altered between human LSCCs and paired normal tissues. Potential oncogenic functions of one such gene, DCUN1D5, were further characterized in vitro. Results: Our results demonstrated that DCUN1D5 was highly expressed in LSCCs. Overexpression of DCUN1D5 in vitro resulted in 2.7-fold increased cellular migration, 67.5% increased invasive capacity, and 2.6-fold increased proliferation. Endogenous DCUN1D5 expression was decreased in a time-dependent manner after genotoxic stress, and silencing of DCUN1D5 by siRNA decreased the number of cells in the S phase by 10.2% and increased apoptosis by 11.7%. Conclusion: Our data suggest that DCUN1D5 in vitro might have vital roles in DNA damage response, but further studies are warranted to assess its significance in vivo.

Drug Resistance Effects of Ribosomal Protein L24 Overexpression in Hepatocellular Carcinoma HepG2 Cells

Guo, Yong-Li,Kong, Qing-Sheng,Liu, Hong-Sheng,Tan, Wen-Bin Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.22

Background: The morbidity and mortality rate of liver cancer continues to rise in China and advanced cases respond poorly to chemotherapy. Ribosomal protein L24 has been reported to be a potential therapeutic target whose depletion or acetylation inhibits polysome assembly and cell growth of cancer. Materials and Methods: Total RNA of cultured amycin-resistant and susceptible HepG2 cells was isolated, and real time quantitative RT-PCR were used to indicate differences between amycin-resistant and susceptible strains of HepG2 cells. Viability assays were used to determine amycin resistance in RPL24 transfected and control vector and null-transfected HepG2 cell lines. Results: The ribosomal protein L24 transcription level was 7.7 times higher in the drug-resistant HepG2 cells as compared to susceptible cells on quantitative RT-PCR analysis. This was associated with enhanced drug resistance as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions: The ribosomal protein L24 gene may have effects on drug resistance mechanisms in hepatocellular carcinoma HepG2 cells.

Astilbin alleviates sepsis-induced acute lung injury by inhibiting the expression of macrophage inhibitory factor in rats

Hong-bo Zhang,Li-chao Sun,Li-da Zhi,Qian-kuan Wen,Zhi-wei Qi,Sheng-tao Yan,Wen Li,Guo-qiang Zhang 대한약학회 2017 Archives of Pharmacal Research Vol.40 No.10

Sepsis is a systemic inflammatory responsesyndrome caused by severe infections. Astilbin is a dihydroflavonolderivative found in many medicinal and foodplants with multiple pharmacological functions. To investigatethe effects of astilbin on sepsis-induced acute lunginjury (ALI), cecal ligation and puncture was performed onrats to establish a sepsis-induced ALI model; these ratswere then treated with astilbin at different concentrations. Lung injury scores, including lung wet/dry ratio, proteinleakage, myeloperoxidase activity, and inflammatory cellinfiltration were determined to evaluate the effects ofastilbin on sepsis-induced ALI. We found that astilbintreatment significantly attenuates sepsis-induced lunginjury and improves survival rate, lung injury scores, lungwet/dry ratio, protein leakage, myeloperoxidase activity,and inflammatory cell infiltration. Astilbin treatment alsodramatically decreased the production of inflammatorycytokines and chemokines in bronchoalveolar lavage fluid. Further, astilbin treatment inhibited the expression andproduction of macrophage inhibitory factor (MIF), whichinhibits the inflammatory response. Collectively, these datasuggest that astilbin has a protective effect against sepsisinducedALI by inhibiting MIF-mediated inflammatoryresponses. This study provides a molecular basis for astilbinas a new medical treatment for sepsis-induced ALI.

Enzymatic Synthesis of Theanine with Escherichia coli γ-glutamyltranspeptidase from a Series of γ-glutamyl Anilide Substrate Analogues

Hong-juan Zhang,Wei-guo Zhang,Zhi-yuan Wang,Yue-ping Zhan,Li-sheng Xu,Jun-zhong Liu,Qian Liu,Qing-cai Jiao 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2

In order to investigate the catalytic mechanism of Escherichia coli γ-glutamyltranspeptidase, ten para- and meta- substituted γ-glutamyl anilides were chemically prepared and employed as substrates to synthesize L-theanine to assay the activity of γ-glutamyltranspeptidase. The reaction was optimized for γ-glutamyl-p-nitroanilide. Key factors such as substrate specificity, pH, temperature, and the substrate mole ratio were all investigated. Kinetic studies of the acyl transfer reaction were described and the Hammett plot was constructed. This study indicated that the ratelimiting acylation reaction of γ-glutamyltranspeptidase can apparently be accelerated by either the electron-withdrawing or electron-donating substituents of γ-glutamyl anilides. The reaction could be catalyzed by the general acid and carboxy of Asp-433 or phenolic hydroxyl Tyr-444 may be the acid by autodock simulation for all prepared γ-glutamyl anilides.

Transgenic Rice Overexperessing a Tomato Mitochondrial Phosphate Transporter, SlMPT3;1, Promotes Phosphate Uptake and Increases Grain Yield

Xian-guo Cheng,Guo-hong Yu,Sheng-cai Huang,Rui He,Ying-Zhang Li 한국식물학회 2018 Journal of Plant Biology Vol.61 No.6

Mitochondrial phosphate transporter plays animportant regulatory role in promoting the uptake andtransport of phosphate in plants. In this study, the SlMPT3;1gene, a member of mitochondrial phosphate transporterfamily in tomato, was isolated and transformed into the riceOryza sativa L. ssp. japonica cultivar Kitaake. The SlMPT3;1 islocalized to the mitochondrial membrane and functions incompensating the phosphate uptake in yeast MB192 mutantthat is defective in phosphate transport under Pi deficiency. RT-qPCR showed that the SlMPT3;1 is expressed in all oftomato tissues, but highly accumulated in the young leavesand stems under Pi deficiency. The data demonstrated that atleast two copies of the SlMPT3;1 gene are inserted into therice genome, and the transcripts of the SlMPT3;1 mRNA arehighly accumulated in the roots of the transgenic rice. Theoverexpression of the SlMPT3;1 gene not only promotesphosphate uptake by the roots, but also increases thetranslocation of phosphate from the roots to the shoots in thetransgenic rice. The transgenic rice accumulated morechlorophyll and soluble sugar in the shoots than the wildtype under Pi deficiency. Microassay sequencing showedthat the differentially expressed genes in the transgenic riceare mainly involved in the regulations of biological processand molecular function under Pi deficiency. Further RTqPCRanalyses revealed that the differentially expressedgenes, which are involved in the regulations of the biologicalprocess, cell component, and molecular function, are upregulatedunder Pi deficiency, and exhibit similar expressiontrends to the relative expression folds of these partial differentiallyexpressed genes in the transcriptomic analyses. This studysuggests that the overexpression of the SlMPT3;1 gene promotedthe uptake and transport of phosphate in rice, thus leading to anenhanced increase in tiller number and effective panicle of perplant, and increasing grain yield under Pi deficiency.

Correlation between Non-Alcoholic Fatty Liver Disease and Visceral Adipose Tissue in Non-Obese Chinese Adults: A CT Evaluation

Ai-Hong Yu,Yang-Yang Duan-Mu,Yong Zhang,Ling Wang,Zhe Guo,Yong-Qiang Yu,Yu-Sheng Wang,Xiao-Guang Cheng 대한영상의학회 2018 Korean Journal of Radiology Vol.19 No.5

Objective: To investigate the correlation between non-alcoholic fatty liver disease and visceral adipose tissue in non-obese Chinese adults using computed tomography (CT). Materials and Methods: The study included 454 subjects undergoing abdominal CT scan. Degree of CT attenuation in liver and spleen, and the degree of fat infiltration in liver were evaluated according to three indices: the attenuation value of liver parenchyma (CTLP), the attenuation ratio of liver and spleen (LSratio) and the attenuation difference between liver and spleen (LSdif). Visceral fat area (VFA) and total fat area (TFA) at L2/3 and L4/5 levels were measured, and the abdominal subcutaneous fat area (SFA) was calculated. Bivariate correlation analysis was carried out to determine the correlation among these factors. Results: In men, VFA, SFA and TFA at L2/3 and L4/5 levels showed significant differences in terms of the three indices to distinguish fatty liver from non-fatty liver (all, p < 0.001). In men, all the three indices showed negative correlation with TFA, SFA and VFA (all, p < 0.001). The negative correlation between the three indices and VFA at the L2/3 level was higher than at L4/5 level (r = -0.476 vs. r = -0.340 for CTLP, r = -0.502 vs. r = -0.413 for LSratio, r = -0.543 vs. r = -0.422 for LSdif, p < 0.001, respectively). The negative correlation between LSratio, LSdif and VFA at L2/3 and L4/5 levels was higher than SFA at the corresponding level. In women, all the three indices showed negative correlation with VFA and TFA at L2/3 and L4/5 levels, and the negative correlation between CTLP and VFA was higher at L2/3 level than at L4/5 level (r = -0.294 vs. r = -0.254, p < 0.001). Conclusion: In non-obese Chinese adults, the degree of hepatic fatty infiltration showed a strong correlation with abdominal fat on CT. VFA at L2/3 level was more closely related to fatty liver compared with VFA at L4/5 level.

Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

( Ai Sheng Xiong ),( Quan Hong Yao ),( Ri He Peng ),( Xian Li ),( Hui Qin Fan ),( Mei Jin Guo ),( Si Liang Zhang ) 생화학분자생물학회 2004 BMB Reports Vol.37 No.3

Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyII) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyII and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyII and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyIIs, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pustoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of I? pustoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that -4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and anoptimum temperature of 60℃.

Directed Evolution of Beta-galactosidase from Escherichia coil into Beta-glucuronidase

( Ai Sheng Xiong ),( Ri He Peng ),( Jing Zhuang ),( Jin Ge Liu ),( Fang Xu ),( Bin Cai ),( Zhao Kui Guo ),( Yu Shan Qiao ),( Jian Min Chen ),( Zhen Zhang ),( Quan Hong Yao ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.3

Photoluminescent Properties of Eu(III) in the Composite Heterocyclic Ligands/Crown Ether Systems

Liu, Hong Guo,Jang, Ki-Wan,Feng, Xu Sheng,Kim, Chang-Dae,Yoo, Young-Jae,Lee, Yong-Ill Korean Chemical Society 2005 Bulletin of the Korean Chemical Society Vol.26 No.12

Composite systems of $Eu(phen)_2Cl_3{(H_2O)}_2$, Eu(DN-bpy)$(phen)Cl_3{(H_2O)}_2$ and Eu(DB-bpy)$(phen)Cl_3{(H_2O)}_2$ (DNbpy: $4,4^\prime$-Dinonyl-$2,2^\prime$-dipyridyl; DB-bpy: $4,4^\prime$-Di-tert-butyl-$2,2^\prime$-dipyridyl) with crown ethers of Benzo-15-crown-5 (B15C5), Benzo-18-crown-6 (B18C6), 18-crown-6 (18C6), Dibenzo-18-crown-6 (DB18C6) and Dibenzo-24-crown-8 (DB24C8) were fabricated successfully and characterized by using photoluminescent spectroscopy and luminescent lifetime measurements. All composites formed show high luminescence mainly in red region. It was found that the heterocyclic ligands such as phen, DN-bpy and DB-bpy as well as the crown ethers have great influences on the photoluminescent properties of $Eu^{3+}$ ion. The environment around $Eu^{3+}$ ion in the composite systems changes greatly,presumably the variation of the first coordination sphere. The $Eu^{3+}$ ion occupies higher symmetrical environment and in more than one kind of symmetrical site in the composite systems studied in this work.