Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum)

Zhongwei Gan,Jinkui Yang,Nan Tao,Zefen Yu,Ke-Qin Zhang 한국미생물학회 2007 The journal of microbiology Vol.45 No.5

Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5'-SYGGRG-3') and nitrogen (5'-GATA-3') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor.

miR-139-5p promotes neovascularization in diabetic retinopathy by regulating the phosphatase and tensin homolog

Zhongwei Zhang,Caiping Song,Tao Wang,Lei Sun,Ling Qin,Jianghua Ju 대한약학회 2021 Archives of Pharmacal Research Vol.44 No.2

Pathological retinal neovascularization is adriver of the progression of diabetic retinopathy (DR). Thepresent study sought to identify the microRNAs (miRNAs)that are diff erentially expressed during the progression ofDR as well as to explore the specifi c regulatory mechanismof those miRNAs in retinal neovascularization. Using amicroarray data set and a diabetic mouse model, it was determinedthat miR-139-5p was signifi cantly upregulated duringthe progression of DR. The in vitro investigation revealedan elevation in the miR-139-5p level in both the high glucose(HG)-treated mouse retinal microvascular endothelialcells (mRMECs) and the HG-treated human RMECs(hRMECs). The miR-139-5p overexpression elevated cellmigration, facilitated tube formation, and increased vascularendothelial growth factor (VEGF) protein level in thehRMECs. While the angiogenic eff ect of miR-139-5p overexpressionwas halted by an anti-VEGF antibody. Meanwhile,the miR-139-5p knockdown eliminated the VEGFinducedcell migration and tube formation in the hRMECs. The phosphatase and tensin homolog (PTEN) was the targetgene of the miR-139-5p. PTEN overexpression removed theangiogenic eff ect of miR-139-5p overexpression, which ledto reduced cell migration and tube formation. In the diabeticmice, the miR-139-5p antagomir eff ectively decreased theacellular capillaries and suppressed the formation of aberrantblood vessels in the retinal tissues. Taken together, miR-139-5p promotes retinal neovascularization by repressingPTEN expression.

Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum)

Gan, Zhongwei,Yang, Jinkui,Tao, Nan,Yu, Zefen,Zhang, Ke-Qin The Microbiological Society of Korea 2007 The journal of microbiology Vol.45 No.5

Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5'-SYGGRG-3') and nitrogen (5'-GATA-3') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor.

circLETM1 upregulates KRT80 via adsorbing miR-143-3p and promotes the progression of colorectal cancer

Li Hua,Guo Junyu,Qin Zhongwei,Wei Mingwei,Guo Houji,Huang Fuda 대한독성 유전단백체 학회 2023 Molecular & cellular toxicology Vol.19 No.3

Background Emerging evidence has indicated that circular RNAs (circRNAs) play vital roles in colorectal cancer (CRC). CircRNA leucine zipper and EF-hand containing transmembrane protein 1 (circLETM1) have been found to be ectopic in CRC, but their role in CRC has not been studied yet. Objective This study aims to explore the roles of circLETM1 in CRC and the possible molecular mechanism. Result The results showed that circLETM1 was highly expressed in CRC cells, and was localized at the cytoplasm. Functional assays verifi ed that overexpression of circLETM1 could facilitate, but circLETM1 downregulation could hamper proliferation, migration and invasion abilities of CRC cells. In addition, mechanism assays found that circLETM1 absorbed and reduced microRNA (miR)-143-3p expression to promote keratin 80 (KRT80). Rescue experiments proved that circLETM1 mediated CRC proliferation, migration and invasion through KRT80. Conclusion Taken together, our results suggested that circLETM1 promotes proliferation, migration and invasion through the miR-143-3p/KRT80 axis in CRC. Therefore, targeting circLETM1 may provide a novel option for CRC treatment.

Fabrication of hydrophobic ZIFs based composite membrane with high CO2 absorption performance

Li Xu,Yu Qin,Liying Liu,Juntian Xiao,Zhongwei Ding 한국화학공학회 2021 Korean Journal of Chemical Engineering Vol.38 No.5

The membrane contactor has been considered an effective device to capture CO2, but its performance is limited by membrane wetting phenomena. In this work, the ZIF-8/ZIF-L (zeolitic imidazolate framework) based composite membrane with excellent hydrophobic stability and CO2 permeability was successfully fabricated by coating the dense layer on the porous PVDF (polyvinylidene fluoride) substrate. The dense layer was developed by evenly incorporating ZIF-8 nanocrystals and ZIF-L nanosheets into the polydimethylsiloxane (PDMS) polymer. The as-prepared membrane possessed competitive hydrophobicity with a water contact angle (WCA) of 131.8o. And after a ten-day immersion test in CO2 absorbent, the WCA only declined by 9.8%, exhibiting excellent hydrophobic stability. Besides, the CO2 permeability of the composite membrane was 61.8×10-8·mol·m-2·s-1·Pa-1 with an ideal CO2/N2 separation factor of ~20, which was effectively improved in comparison with the control membrane. In the CO2 absorption experiment, the prepared membrane achieved the competitive CO2 flux of 3.02×10-3·mol·m-2·s-1 when the liquid velocity was 0.25m·s-1 and displayed robust long-term operating stability. Finally, the AC impedance technique was applied to continuously monitor the wetting state of composite membranes, which further investigated their anti-wetting ability.

Effects of Glucagon-Like Peptide-2-Expressing Saccharomyces cerevisiae Not Different from Empty Vector

( Xi Zhong ),( Guopeng Liang ),( Lili Cao ),( Qi Qiao ),( Zhi Hu ),( Min Fu ),( Hong Bo ),( Qin Wu ),( Guanlin Liang ),( Zhongwei Zhang ),( Lin Zhou ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.10

Saccharomyces cerevisiae (S. cerevisiae) and glucagon-like peptide-2 (GLP-2) have been employed to improve the intestinal development of weaned animals. The goal of this study was to determine whether either exogenous S. cerevisiae or GLP-2 elicits major effects on fecal microbiotas and cytokine responses in weaned piglets. Ninety-six piglets weaned at 26 days were assigned to one of four groups: 1) Basal diet (Control), 2) empty vector-harboring S. cerevisiae (EV-SC), 3) GLP-2-expressing S. cerevisiae (GLP2-SC), and 4) recombinant human GLP-2 (rh-GLP2). At the start of the post-weaning period (day 0), and at day 28, fecal samples were collected to assess the bacterial communities via sequencing the V1-V2 region of the 16SrRNA gene, and piglets’ blood was also sampled to measure cytokine responses (i.e., IL-1β, TNF-α, and IFN-γ). This study revealed that, on the one hand, although S. cerevisiae supplementation did not significantly alter the growth of weaned piglets, it induced increases in the relative abundances of two core genera (Ruminococcaceae_norank and Erysipelotrichaceae_norank) and decreases in the relative abundances of two other core genera (Lachnospiraceae_norank and Clostridiale_norank) and cytokine levels (IL-1β and TNF-α) (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). On the other hand, GLP-2 supplementation had no significant influence on fecal bacterial communities and cytokine levels, but it produced better body weight and average daily gain (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). Therefore, altered fecal microbiotas and cytokine response effects in weaned piglets were due to S. cerevisiae rather than GLP-2.