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 comparative study on population development patterns of Sogatella furcifera between tropical and subtropical areas

Yang Hu,Jia-An CHENG,Zengrong Zhu,Kong Luen Heong,Qiang Fu,Jiachun He 한국응용곤충학회 2014 Journal of Asia-Pacific Entomology Vol.17 No.4

The White-backed Planthopper (WBPH), Sogatella furcifera (Horváth (Hemiptera: Delphacidae)) has been the most serious pest threatening rice production in Asia since the late 1970s. A series of field experiments using the same research protocol was carried out to compare the effects of main environmental factors on population development of WBPH in tropical (Philippines) and subtropical (China) areas in 2010–2012 and to provide further evidences of ecological mechanisms involved that cause frequent outbreak in subtropical rice. Outcomes showed that WBPH population in subtropical area could be characterized as the higher population growth rates and higher peak densities. The average growth rate (116.60 ± 46.16) in subtropical area was significantly higher than that in tropical area (24.02± 11.25). The higher realized fecundity in subtropical area indicated that the higher growth rates and higher peak densities were mainly related to the poor natural regulating forces in subtropical area. Our results showed that resistant variety could significantly reduce the peak density in subtropical areas, but not in tropical areas. We inferred that the reason for not detecting the effect of resistant variety in tropical area was due to the dominant controlling effects provided by natural enemies,which means that the natural regulating effect in tropical areawas strong enough to disguise the effects of resistant variety. The significant interactions between plant resistance and location demonstrated that integration of natural enemies and use of plant resistance could play important roles for reducing outbreak frequency of WBPH effectively in subtropical rice.

Branched polymeric prodrug/programmed cell death 4 complexes for combinational cancer therapy

Yu‑Jing He,Lei Xing,Peng‑Fei Cui,Jia‑Liang Zhang,Jian‑Bin Qiao,Cheng‑Qiong Luo,Ge Jiang,Hu‑Lin Jiang 한국약제학회 2017 Journal of Pharmaceutical Investigation Vol.47 No.2

Here, we demonstrate a co-delivery system constructed by integrating chemotherapeutic molecules into branched polymeric prodrug which can condense nucleic acids. Demethylcantharidate (DCA) was chosen as a model drug and premodified through nucleophilic substitution reaction by its two carboxylic groups with allyl chloride. The synthesized intermediate (DCA-dially) was then used to polymerize with tris (2-aminoethyl) amine (TAEA) through progressively ammonolysis reaction. The obtained poly (DCA-alt-TAEA) (DCAT) was used to pack PDCD4 into spherical-like nanoparticles through electrostatic interaction. Gel retardation assays implied that DCAT could integrate DNA at the weight ratio of 1 and protect it from digestion by nuclease. Acid-base titration experiments showed that DCAT obtained preferable buffering capability which was beneficial for the endosomal escape of DCAT/PDCD4 complexes. Cellular tests involving gene transfection efficiency and cytotoxicity indicated that DCA and PDCD4 co-delivered by the complexes significantly and synergistically suppressed the viability of SMMC-7721 cells. These results suggest that integrating chemotherapeutic molecules into nucleic acid-packing polymeric prodrug as cationic polymer/PDCD4 complexes is a highly efficient way to co-deliver chemotherapeutic drugs and nucleic acids for cancer therapy.

MiR-214 Regulates the Human Hair Follicle Stem Cell Proliferation and Differentiation by Targeting EZH2 and Wnt/b- Catenin Signaling Way In Vitro

Ke-Tao Du,Jia-Qin Deng,Xu-Guang He,Zhao-ping Liu,Cheng Peng,Mingsheng Zhang 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.3

miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, b-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/b-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/b-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR- 214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, b-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, b-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of b-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR- 214 directly targets EZH2 and affects Wnt/b-catenin signaling. The miR-214/EZH2/b-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.

Low Counts of γδ T Cells in Peritumoral Liver Tissue are Related to More Frequent Recurrence in Patients with Hepatocellular Carcinoma after Curative Resection

Cai, Xiao-Yan,Wang, Jia-Xing,Yi, Yong,He, Hong-Wei,Ni, Xiao-Chun,Zhou, Jian,Cheng, Yun-Feng,Jin, Jian-Jun,Fan, Jia,Qiu, Shuang-Jian Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.2

Objectives: TCR-gamma-delta+T cells (${\gamma}{\delta}$ T cells) are non-conventional T lymphocytes that can recognize and eradicate tumor cells. Our previous studies showed that infiltration and function of ${\gamma}{\delta}$ T cells were substantially attenuated in hepatocellular carcinoma (HCC). However, their prognostic value was not clarified. Methods: The association between ${\gamma}{\delta}$ T cells and the clinical outcomes was determined by immunohistochemistry (IHC) in a HCC patient cohort (n = 342). Results:Immunohistochemistry showed decreased infiltration of ${\gamma}{\delta}$ T cells in tumoral tissues compared with paired peritumoral tissues. The counts of ${\gamma}{\delta}$ T cells in peritumoral tissues were negatively correlated with tumor size (P = 0.005). Survival analysis showed that the levels of peritumoral ${\gamma}{\delta}$T cells were related to both time to recurrence (TTR) and overall survival (OS) (P = 0.010 and P = 0.036, respectively) in univariate analysis, and related to TTR in multivariate analysis (P = 0.014, H.R. [95% CI] = 0.682 [0.502-0.927]). Furthermore, the level of peritumoral ${\gamma}{\delta}$ T cells showed independent prognostic value for TTR in Barcelona Clinic Liver Cancer (BCLC) stage A patients (P = 0.038, H.R. [95% CI] = 0.727 [0.537-0.984]). However, tumoral ${\gamma}{\delta}$ T cells did not show independent prognostic value for either TTR or OS in HCC patients. Conclusions: Low counts of ${\gamma}{\delta}$ T cells in peritumoral liver tissue are related to a higher incidence of recurrence in HCC and can predict postoperative recurrence, especially in those with early-stage HCC.

Effects of Ginsenosides Rg1 on Osteoblasts Cultured with Ti Particles

( Yu Lin ),( Yin Sheng Wu ),( Jia Cheng He ),( Yun Mei Huang ),( Yan Ping Lin ) 한국응용약물학회 2012 Biomolecules & Therapeutics(구 응용약물학회지) Vol.20 No.1

The aim of this study was to explore the role and effect of ginsenosides Rg1 on osteoblasts cultured with Ti particles. Osteoblasts from neonatal rats were cultured with particles and different doses of Rg1, the main active ingredient in ginsenosides Rg1. We found that the COX-2, PGE2, TNF-α, IL-1, and IL -6 concentrations in the medium of cells cultured with Ti particles significantly increased as compared with that of the control cells (p<0.05 or p<0.01). In addition, cells cultured with Ti particles alone exhibited the highest concentrations of these molecules. The PGE2, TNF-α, IL-1, and IL-6 levels in the medium of cells cultured with Rg1 were in between those of the control cells and the cells cultured with Ti particles alone. The IL-1ra level in the group cultured with Ti and medium-dose Rg1 was the highest followed by the cells cultured with Ti and high-dose Rg1 and those cultured with Ti and low-dose Rg1 (p<0.05). In conclusion, ginsenosides can reduce the levels of inflammatory cytokines produced by osteoblasts on induction with Ti particles and can prevent prosthesis loosening.

Recombinant-attenuated Salmonella Pullorum strain expressing the hemagglutinin-neuraminidase protein of Newcastle disease virus (NDV) protects chickens against NDV and Salmonella Pullorum challenge

Ke Ding,Ke Shang,Zu-Hua Yu,Chuan Yu,Yan-Yan Jia,Lei He,Cheng-Shui Liao,Jing Li,Chun-Jie Zhang,Yin-Ju Li,Ting-Cai Wu,Xiang-chao Cheng 대한수의학회 2018 Journal of Veterinary Science Vol.19 No.2

Newcastle disease virus (NDV) and Salmonella Pullorum have significant damaging effects on the poultry industry, but no previous vaccinecan protect poultry effectively. In this study, a recombinant-attenuated S. Pullorum strain secreting the NDV hemagglutinin-neuraminidase(HN) protein, C79-13ΔcrpΔasd (pYA-HN), was constructed by using the suicide plasmid pREasd-mediated bacteria homologousrecombination method to form a new bivalent vaccine candidate against Newcastle disease (ND) and S. Pullorum disease (PD). The effectof this vaccine candidate was compared with those of the NDV LaSota and C79-13ΔcrpΔasd (pYA) strains. The serum hemagglutinationinhibition antibody titers, serum immunoglobulin G (IgG) antibodies, secretory IgA, and stimulation index in lymphocyte proliferation wereincreased significantly more (p < 0.01) in chickens inoculated with C79-13ΔcrpΔasd (pYA-HN) than with C79-13ΔcrpΔasd (pYA) but werenot significantly increased compared with the chickens immunized with the LaSota live vaccine (p > 0.05). Moreover, the novel strain provides60% and 80% protective efficacy against the NDV virulent strain F48E9 and the S. Pullorum virulent strain C79-13. In summary, in this study,a recombinant-attenuated S. Pullorum strain secreting NDV HN protein was constructed. The generation of the S. Pullorum C79-13ΔcrpΔasd(pYA-HN) strain provides a foundation for the development of an effective living-vector double vaccine against ND and PD.

Microscopic Mechanisms and Acoustic Emission Characteristics of Sandy Mudstone under Different Water Saturations

Xiao-ming Sun,Fu-kun Shi,Ming-qun Zhu,Jia-xu Ding,Lin-sen He,Zhi-hu Li,Tong Zhang,Cheng-yu Miao 대한토목학회 2024 KSCE Journal of Civil Engineering Vol.28 No.1

In order to study the damage mechanism of surrounding rocks under water-rock coupling, an experimental study on the microscopic law of water absorption and mechanics of sandy mudstone was carried out in the sandy mudstone stratum of Wanfu coal mine in Shandong, China. The results indicate that the strength of sandy mudstone exhibits three stages of relative softening, rapid softening, and decelerated softening. The speed of water molecules entering the microscopic pore structure was analyzed based on the percentage of water molecules in the pore structure. The acoustic emission energy release process is divided into energy accumulation stage, stable release stage and intense release stage. With the increase of water content saturation,  (Spectral width of the multifractal spectrum) tends to decrease and the distribution of events is gradually uniform. The action of water gradually expands and penetrates the cracks and pores inside the sandy mudstone, which makes f (Frequency of occurrence of the signal) change from negative to positive values.

Complete ¹H-NMR and <SUP>13</SUP>C-NMR spectral assignment of five malonyl ginsenosides from the fresh flower buds of Panax ginseng

Yu-Shuai Wang,Yin-Ping Jin,Wei Gao,Sheng-Yuan Xiao,Yu-Wei Zhang,Pei-He Zheng,Jia Wang,Jun-Xia Liu,Cheng-He Sun,Ying-Ping Wang 고려인삼학회 2016 Journal of Ginseng Research Vol.40 No.3

Background: Ginsenosides are the major effective ingredients responsible for the pharmacological effects of ginseng. Malonyl ginsenosides are natural ginsenosides that contain a malonyl group attached to a glucose unit of the corresponding neutral ginsenosides. Methods: Medium-pressure liquid chromatography and semipreparative high-performance liquid chromatography were used to isolate purified compounds and their structures determined by extensive one-dimensional- and two-dimensional nuclear magnetic resonance (NMR) experiments. Results: A new saponin, namely malonyl-ginsenoside Re, was isolated from the fresh flower buds of Panax ginseng, along with malonyl-ginsenosides Rb1, Rb2, Rc, Rd. Some assignments for previously published ¹H- and <SUP>13</SUP>C-NMR spectra were found to be inaccurate. Conclusion: This study reports the complete NMR assignment of malonyl-ginsenoside Re, Rb₁, Rb₂, Rc, and Rd for the first time.