Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells

Yong-Shi Liu,Qiong Liu,Yanlong Jiang,Wentao Yang,Hai-Bin Huang,Chun-Wei Shi,Gui-Lian Yang,Chun-Feng Wang 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.4

Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN- λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.

MicroRNA-27a Inhibits Cell Migration and Invasion of Fibroblast-Like Synoviocytes by Targeting Follistatin-Like Protein 1 in Rheumatoid Arthritis

Shi, Dong-liang,Shi, Gui-rong,Xie, Jing,Du, Xu-zhao,Yang, Hao Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.8

Fibroblast-like synoviocytes (FLS) with aberrant expression of microRNA (miRNA) are critical pathogenic regulators in rheumatoid arthritis (RA). Previous studies have found that overexpression or silencing of miRNA can contribute to the development of miRNA-based therapeutics in arthritis models. In this study, we explored the effects of miR-27a on cell migration and invasion in cultured FLS from RA patients. We found that miR-27a was markedly downregulated in the serum, synovial tissue, and FLS of RA patients. Meanwhile, the expression of follistatin-like protein 1 (FSTL1) was upregulated, which suggests that FSTL1 plays a key role in RA development. The results of a Transwell assay showed that miR-27a inhibited FLS migration and invasion. However, miR-27a inhibition promoted the migration and invasion of FLS. In addition, the down-regulated expression of matrix metalloproteinases (MMP2, MMP9, and MMP13) and Rho family proteins (Rac1, Cdc42, and RhoA) was detected after treatment with miR-27a in RA-FLS by quantitative reverse transcription-PCR and western blot analysis. Then, a luciferase reporter assay validated that miR-27a targeted the 3-untranslated region (3'-UTR) of FSTL1. Moreover, miR-27a caused a significant decrease of FSTL1. In addition, the expression of TLR4 and $NF{\kappa}B$ was inhibited by miR-27a but increased by FSTL1 overexpression. In conclusion, we found that miR-27a inhibited cell migration and invasion of RA-FLS by targeting FSTL1 and restraining the $TLR4/NF{\kappa}B$ pathway.

Genome-wide identification and analysis of MIKC-type MADS-box genes expression in Chimonanthus salicifolius

Gui Fang-Fang,Jiang Ge-Ge,Bin Dong,Zhong Shi-Wei,Xiao Zheng,Qiu Fang,Wang Yi-Guang,Yang Li-Yuan,Zhao Hongbo 한국유전학회 2023 Genes & Genomics Vol.45 No.9

Background MIKC type MADS-box transcription factors are one of the largest gene families and play a pivotal role in flowering time and flower development. Chimonanthus salicifolius belongs to the family Calycanthaceae and has a unique flowering time and flowering morphology compared to other Chimonanthus species, but the research on MIKC type MADS-box gene family of C. salicifolius has not been reported. Objective Identification, comprehensive bioinformatic analysis, the expression pattern of MIKC-type MADS-box gene family from different tissues of C. salicifolius. Methods Genome-wide investigation and expression pattern under different tissues of the MIKC-type MADS-box gene family in C. salicifolius, and their phylogenetic relationships, evolutionary characteristics, gene structure, motif distribution, promoter cis-acting element were performed. Results A total of 29 MIKC-type MADS-box genes were identified from the whole genome sequencing. Interspecies synteny analysis revealed more significant collinearity between C. salicifolius and the magnoliids species compared to eudicots and monocots. MIKC-type MADS-box genes from the same subfamily share similar distribution patterns, gene structure, and expression patterns. Compared with Arabidopsis thaliana, Nymphaea colorata, and Chimonanthus praecox, the FLC genes were absent in C. salicifolius, while the AGL6 subfamily was expanded in C. salicifolius. The selectively expanded promoter (AGL6) and lack of repressor (FLC) genes may explain the earlier flowering in C. salicifolius. The loss of the AP3 homologous gene in C. salicifolius is probably the primary cause of the morphological distinction between C. salicifolius and C. praecox. The csAGL6a gene is specifically expressed in the flowering process and indicates the potential function of promoting flowering. Conclusion This study offers a genome-wide identification and expression profiling of the MIKC-types MADS-box genes in the C. salicifolius, and establishes the foundation for screening flowering development genes and understanding the potential function of the MIKC-types MADS-box genes in the C. salicifolius.

A New Kind of Secure Electronic Communication Technology VT Position Code Communication Technology and Its Implementation

Shi-Ying Zhou,Gui-He Qin,Yang Xun,Yu-Bo Jin 보안공학연구지원센터 2008 International Journal of Security and Its Applicat Vol.2 No.3

Eelectronic device technology has been monopolized by binary system for many years. The circs not only makes it impossible to break through the existing technology bottleneck, but also brings a lot of potential safety problems. Aiming at this question, a new electronic communication technology is presented in this paper. The technology quantifies the time axis and the voltage axis synchronously, uses the quantified time dot as the address of the communication, and realizes the transmission of the multi-system [1] data via transmitting the multi-steps voltage quantification. The technology solves the bottleneck problem of the speed, circuit and electromagnetism in the electronic communication, changes the binary system coding mode and communication connection form of the electronic device, reduces the transmission quantity of the redundant information, advances the security of electronic system and network, debases the complexity of the devices connection, enhances the rate of the processing and the transmission, simplifies the transformation between the difference protocols. The experimental results approve the validity and the robustness of the technology.

Effect of male inflorescence of Castanea mollissima on the reproductive development and lifetime of Bactrocera minax

He Zhangzhang,Gui Lianyou,Wang Fulian,Shi Yongfang,Liang Peng,Yang Xuan,Hua Dengke,Du Tainhua 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.4

Previous work reveals that newly emerged adult Chinese citrus flies, Bactrocera minax (Enderlein), emerges from the ground in the mosaic-type citrus orchard of the hilly terrain landform dispersed into adjacent forest, and the newly emerged adult flies fed on male inflorescence of the Chinese chestnut, Castanea mollissima Blume (Fagaceae). The potential impact of male inflorescence of C. mollissima on the longevity, and survival of adult females and males, the ovarian development of adult females, and the abdominal index, ovarian index, and egg length of the fourth ovarian grade of living females, as well as the flowering process of the male inflorescence of the Chinese chestnut, were studied with the three methods of rearing (normal rearing, hungry rearing, and bagging rearing). For all three rearing treatments, the longevity of all adults could live up to 38 days, there were similar time dynamics for the average survival of female and male adult flies. Of the total females, 32.1%–100% females with bagging rearing could attain the fourth ovarian grade like those in rearing and hunger rearing. The abdominal index, ovarian index, and egg length of the fourth ovarian grade of 27–39 days’ old living females with bagging rearing (it corresponds to the late flower withering period), were 75.1%, 49.8% and 89.6% of those with normal rearing, and 73.2%, 60.6% and 90.8% of those with hungry rearing, respectively. The male inflorescence of C. mollissima can satisfy the nutritional requirements of adult B. minax, enabling their survival and normal reproduction. This study can provide references for development and application of artificial feed and food attractants of adult B. minax.

MicroRNA-27a Inhibits Cell Migration and Invasion of Fibroblast-Like Synoviocytes by Targeting Follistatin-Like Protein 1 in Rheumatoid Arthritis

Dong-liang Shi,Gui-rong Shi,Jing Xie,Xu-zhao Du,Hao Yang 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.8

Fibroblast-like synoviocytes (FLS) with aberrant expres-sion of microRNA (miRNA) are critical pathogenic regula-tors in rheumatoid arthritis (RA). Previous studies have found that overexpression or silencing of miRNA can contribute to the development of miRNA-based therapeutics in arthritis models. In this study, we explored the effects of miR-27a on cell migration and invasion in cultured FLS from RA patients. We found that miR-27a was markedly downregulated in the serum, synovial tissue, and FLS of RA patients. Meanwhile, the expression of follistatin-like protein 1 (FSTL1) was upregulated, which suggests that FSTL1 plays a key role in RA development. The results of a Transwell assay showed that miR-27a inhibited FLS migration and invasion. However, miR-27a inhibition promoted the migration and invasion of FLS. In addition, the down-regulated expression of matrix metalloproteinases (MMP2, MMP9, and MMP13) and Rho family proteins (Rac1, Cdc42, and RhoA) was detected after treatment with miR-27a in RA-FLS by quantitative reverse transcription-PCR and western blot analysis. Then, a luciferase reporter assay validated that miR-27a targeted the 3-untranslated region (3-UTR) of FSTL1. Moreover, miR-27a caused a significant decrease of FSTL1. In addition, the expression of TLR4 and NFκB was inhibited by miR-27a but increased by FSTL1 overexpression. In conclusion, we found that miR-27a inhibited cell migration and invasion of RA-FLS by targeting FSTL1 and restraining the TLR4/NFκB pathway.

Improved antimicrobial effect of ginseng extract by heat transformation

Peng Xue,Yang Yao,Xiu-Shi Yang,Jia Feng,Gui-xing Ren 고려인삼학회 2017 Journal of Ginseng Research Vol.41 No.2

Background: The incidence of halitosis has a prevalence of 22e50% throughout the world and is generally caused by anaerobic oral microorganisms, such as Fusobacterium nucleatum, Clostridium perfringens, and Porphyromonas gingivalis. Previous investigations on the structure-activity relationships of ginsenosides have led to contrasting results. Particularly, the antibacterial activity of less polar ginsenosides against halitosis-related bacteria has not been reported. Methods: Crude saponins extracted from the Panax quinquefolius leaf-stem (AGS) were treated at 130C for 3 h to obtain heat-transformed saponins (HTS). Five ginsenoside-enriched fractions (HTS-1, HTS-2, HTS-3, HTS-4, and HTS-5) and less polar ginsenosides were separated by HP-20 resin absorption and HPLC, and the antimicrobial activity and mechanism were investigated. Results: HPLC with diode-array detection analysis revealed that heat treatment induced an extensive conversion of polar ginsenosides (-Rg1/Re, -Rc, -Rb2, and -Rd) to less polar compounds (-Rg2, -Rg3, -Rg6, -F4, -Rg5, and -Rk1). The antimicrobial assays showed that HTS, HTS-3, and HTS-4 were effective at inhibiting the growth of F. nucleatum, C. perfringens, and P. gingivalis. Ginsenosides-Rg5 showed the best antimicrobial activity against the three bacteria, with the lowest values of minimum inhibitory concentration and minimum bactericidal concentration. One major reason for this result is that less polar ginsenosides can more easily damage membrane integrity. Conclusion: The results indicated that the less polar ginsenoside-enriched fraction from heat transformation can be used as an antibacterial agent to control halitosis.

Differential Impacts on Bacterial Composition and Abundance in Rhizosphere Compartments between Al-Tolerant and Al-Sensitive Soybean Genotypes in Acidic Soil

Wen Zhong-Ling,Yang Min-Kai,Fazal Aliya,Liao Yong-Hui,Cheng Lin-Run,Hua Xiao-Mei,Hu Dong-Qing,Shi Ji-Sen,Yang Rong-Wu,Lu Gui-Hua,Qi Jin-Liang,Zhi Hong,Qian Qiu-Ping,Yang Yong-Hua 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.8

In this study, two soybean genotypes, i.e., aluminum-tolerant Baxi 10 (BX10) and aluminumsensitive Bendi 2 (BD2), were used as plant materials and acidic red soil was used as growth medium. The soil layers from the inside to the outside of the root are: rhizospheric soil after washing (WRH), rhizospheric soil after brushing (BRH) and rhizospheric soil at two sides (SRH), respectively. The rhizosphere bacterial communities were analyzed by high-throughput sequencing of V4 hypervariable regions of 16S rRNA gene amplicons via Illumina MiSeq. The results of alpha diversity analysis showed that the BRH and SRH of BX10 were significantly lower in community richness than that of BD2, while the WRH exhibited no significant difference between BX10 and BD2. Among the three sampling compartments of the same soybean genotype, WRH had the lowest community richness and diversity while showing the highest coverage. Beta diversity analysis results displayed no significant difference for any compartment between the two genotypes, or among the three different sampling compartments for any same soybean genotype. However, the relative abundance of major bacterial taxa, specifically nitrogen-fixing and/or aluminum-tolerant bacteria, was significantly different in the compartments of the BRH and/or SRH at phylum and genus levels, indicating genotype-dependent variations in rhizosphere bacterial communities. Strikingly, as compared with BRH and SRH, the WRH within the same genotype (BX10 or BD2) always had an enrichment effect on rhizosphere bacteria associated with nitrogen fixation

A Newly Isolated Rhizopus microsporus var. chinensis Capable of Secreting Amyloytic Enzymes with Raw-Starch-Digesting Activity

( Yu Na Li ),( Gui Yang Shi ),( Wu Wang ),( Zheng Xiang Wang ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.2

Improved antimicrobial effect of ginseng extract by heat transformation

Xue, Peng,Yao, Yang,Yang, Xiu-shi,Feng, Jia,Ren, Gui-xing The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.2

Background: The incidence of halitosis has a prevalence of 22-50% throughout the world and is generally caused by anaerobic oral microorganisms, such as Fusobacterium nucleatum, Clostridium perfringens, and Porphyromonas gingivalis. Previous investigations on the structure-activity relationships of ginsenosides have led to contrasting results. Particularly, the antibacterial activity of less polar ginsenosides against halitosis-related bacteria has not been reported. Methods: Crude saponins extracted from the Panax quinquefolius leaf-stem (AGS) were treated at $130^{\circ}C$ for 3 h to obtain heat-transformed saponins (HTS). Five ginsenoside-enriched fractions (HTS-1, HTS-2, HTS-3, HTS-4, and HTS-5) and less polar ginsenosides were separated by HP-20 resin absorption and HPLC, and the antimicrobial activity and mechanism were investigated. Results: HPLC with diode-array detection analysis revealed that heat treatment induced an extensive conversion of polar ginsenosides (-Rg1/Re, -Rc, -Rb2, and -Rd) to less polar compounds (-Rg2, -Rg3, -Rg6, -F4, -Rg5, and -Rk1). The antimicrobial assays showed that HTS, HTS-3, and HTS-4 were effective at inhibiting the growth of F. nucleatum, C. perfringens, and P. gingivalis. Ginsenosides-Rg5 showed the best antimicrobial activity against the three bacteria, with the lowest values of minimum inhibitory concentration and minimum bactericidal concentration. One major reason for this result is that less polar ginsenosides can more easily damage membrane integrity. Conclusion: The results indicated that the less polar ginsenoside-enriched fraction from heat transformation can be used as an antibacterial agent to control halitosis.