1.Recombinant Probiotics for Prevention of Enteric Infections
Yong LIN ; Wen YAO ; Wei-Yun ZHU ;
Microbiology 1992;0(06):-
Many microbial pathogens,including those responsible for major enteric infections,exploit oligosaccharides that are displayed on the surface of host cells as receptors for toxins and adhesins. Blocking crucial ligand receptor interactions is therefore a promising therapeutic strategy. One approach is to express molecular mimics of host receptors on the surface of harmless recombinant bacteria that can survive in the gut. These recombinant probiotics bind bacterial toxins in the gut lumen with very high avidity,thereby preventing disease.
2.Kinetic Analysis of the Inhibition of in vitro Growth of Pathogenic Escherichia coli K88 and O138 by Lactobacillus Isolates from Porcine Intestine
Yong LIN ; Wen YAO ; Wei-Yun ZHU ;
Microbiology 2008;0(10):-
Fermentation property and kinetic analysis of the inhibition of in vitro growth of pathogenic Escherichia coli K88, O138 by Lactobacillus intestinalis isolated from porcine intestine mucus were investi-gated. The results of fermentation showed that L1 result in a pH of 3.90 within 12 h and a large amount of lactic acid production, with the concentration of 104.08 mmol/L. Kinetic analysis of the antibacterial activity of L1 metabolic product toward K88, O138 showed that CFUS display strong antibacterial activity toward K88, O138; the antibacterial activity of L1 CFUS was higher than the lactic acid control samples and was mainly due to the production of lactic acid; K88, O138 had the tolerance property toward pH 4.5.
4.RESEARCH PROGRESS ON ENZYMATIC HYDROLYSIS OF NITRILE
Wen-Yong LOU ; Min-Hua ZONG ; Sen-Lin LIU ;
Microbiology 1992;0(06):-
Enzymatic hydrolysis of nitrile was performed under mild conditions and afford high efficiency and selectivity,thus being highly potential for the synthesis of optically active carboxylic acids and their derivatives.The type of nitrile-hydrolyzing enzymes and hydrolysis reactions,reaction characteristics,factors influencing the hydrolysis and the prospect for its application to industrial production were reviewed in this paper.
5.Effect of baicalein on high glucose-induced expression of extracellular matrix and transforming growth factor ?1 in proximal tubular epithelial cells
Wen SHEN ; Fuming LU ; Yong GU ; Shanyan LIN
Chinese Journal of Nephrology 1997;0(03):-
Objective To investigate the effect of baicalain on high expression of ECM and TGF-?1 in proximal tubular epithelial cells cultured in high glucose concentration. Methods LLC-PK1 cells were divided into six groups: (1)normal glucose group(NG, 5. 5 mmol/L D-glucose), (2)high glucose group(HG, 25 mmol/L D-glucose), (3) HG + PKC inhibitor(10?mol/L chelerythrine chloride), (4)HG + baicalein(50 ?mol/L), (5) HG + baicalein(100 ?mol/L), (6) HG + baicalein (200 ?mol/L) . PKC activity was detected. Expression of ColⅣ, FN and TGF-?1 was examined by in situ hybridization and immunocytochemistry(ABC) techniques. Results At concentrations of 100 ?mol/L and 200 ?mol/L, baicalein decreased membranous PKC activity in LLC-PK1 cells by 42% and 68% , respectively ( P
6.Effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.
Guo-Lin WU ; Tian-Yi LI ; Wen-Wen LU ; Guo-You YU ; Yong-Sheng FAN
China Journal of Chinese Materia Medica 2013;38(23):4148-4151
OBJECTIVETo observe the effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.
METHODThirty-two NOD mice were randomly divided into the model group, the traditional Chinese medicine group (TCM group, orally given 0.4 mL nourishing Yin, strengthening Qi and activating blood decoction as per 100 g x kg(-1) everyday), the hydroxychloroquine group (given 0.4 mL hydroxychloroquine as per 60 mg x kg(-1) everyday), the traditional Chinese medicine and western medicine group (TCM WM group, given nourishing Yin, Strengthening Qi and activating blood decoction 50 g x kg(-1) and hydroxychloroquine 60 mg x kg(-1), 0.4 mL everyday), with eight mice in each group. Eight Balb/C mice were selected as the normal control group (normal group). All of mice were killed after eight weeks, and their submaxillary glands were dissected. The expression levels of Fas/FasL were examined by immunohistochemical method, and the FasL mRNA was detected by RT-PCR.
RESULTThe expression levels of Fas/FasL in salivary glands of the model group were higher than that of other groups (P < 0.05). The expression level of FasL of the normal group was much lower than that in the hydroxychloroquine group (P < 0.05). The relative expression level of Fas mRNA in salivary glands of the model group was higher than that in other groups, but the control group was notably lower than other groups (P < 0.05). The expression level of FasL mRNA in salivary glands of the model group was higher than that in TCM and TCM WM groups (P < 0.05). But the expression level in TCM WM group was notably lower than the hydroxychloroquine group (P < 0.05).
CONCLUSIONThe nourishing Yin, strengthening Qi and activating blood decoction could down-regulate the expression level of Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression, and had a better efficacy after being combined with hydroxychloroquine. The nourishing Yin, strengthening Qi and activating blood decoction might treat the Sjogren's Syndrome by reducing apoptosis which is regulated by Fas/FasL
Animals ; Fas Ligand Protein ; genetics ; Female ; Gene Expression Regulation ; Medicine, Chinese Traditional ; methods ; Mice ; Mice, Inbred NOD ; Qi ; RNA, Messenger ; genetics ; metabolism ; Salivary Glands ; metabolism ; Sjogren's Syndrome ; blood ; genetics ; therapy ; Yin-Yang ; fas Receptor ; genetics
7.Construction,Identification and Expression of Recombinant Eukaryotic Vector pCAG-IRES-SHIP-GFP on Porliferation of Leukemia Cell Line K562
Lin YANG ; Jian-Min LUO ; Xiao-Jun LIU ; Zhi-Yong CHENG ; Shu-Peng WEN ; Xiao-Yang YANG ; Xue-Wen WU ;
China Biotechnology 2006;0(06):-
The aim was to construct and identify the mammalian expression vector of pCAG-IRES-SHIP-GFP and to detect whether it could express in human acute leukemia cell line K562.The cDNA fragment of SHIP obtained by RT-PCR was inserted into pCAG-IRES-GFP.The recombinant plasmid was confirmed by restriction enzyme digesiton,PCR and DNA sequecing.pCAG-IRES-SHIP-GFP was transfected into K562 cells with lipofectamine 2000.The expression of SHIP was determined by GFP fluorescence and Western blot analysis.FQ-PCR was used to quantitate SHIP mRNA.The expression of p-Akt,Akt were determined by Western blot.PI were tested by flow cytometry and MTT to verify whether exogenous SHIP could inhibit proliferation of K562 cells.The results showed that the correct constrution of the recombinant plasmid pCAG-IRES-SHIP-GFP has been shown by restriction enzyme digestion,PCR and DNA sequencing.pCAG-IRES-SHIP-GFP could express SHIP protein in K562 cells.The K562 cells viability after transfected with SHIP gene droped down.Western blot analysis showed that phospha-Akt308 and Akt473 were reduced to 38.7% and 68% respectively.It was concluded that the vector of pCAG-IRES-SHIP-GFP has been successfully constructed and it can be expressed in K562 cells.The expression of exogenous SHIP gene can lead to apoptosis of K562 cells by down-regulating the p-Akt expression.What found here might be one of the mechanisms involved in the pathogenesis of leukemia.
8.Minocycline protects dopaminergic neurons in lipopolysaccharide.induced model of Parkinson' s disease
Qin-Yong YE ; Hai-Hua YANG ; Ping-Yi XU ; Zhuo-Lin LIU ; Hao-Wen XU ; Wei-Wen ZHU ; An-Mu XIE
Chinese Journal of Neurology 2001;0(02):-
Objective To further investigated the effect of minocycline on the inhibition of microglial activation and subsequent protection of nigral DA neuron.Methods 20 rats injected with LPS in the substantia nigra (SN) were randomly divided into two groups (LPS group and LPS+Minocycline group).The behavior was observed on the 7~(th) d and 14~(th) d.The immunohistoehemistry,in situ hybridization and Western-blot were used to detect the levels of positive neuron,mRNA,protein of TH and OX-42. Results The slightly rotational behavior was observed in LPS+Minoeyeline group.The majority of mieroglias were activated in the two groups.Some microglia in the SNpc remained ramified in LPS+ Minocycline group.The numbers of hypertophie microglia in LPS+Minoeyeline group were less than that in LPS group.Western-blot showed that the protein of OX-42 in two LPS groups was higher than in normal group(P
9.Genetic analysis of hantaviruses carried by Rattus norvegicus collected from Henan and Neimenggu provinces, China
Wen WANG ; Xian-Dan LIN ; Wen-Ping GUO ; Ming-Hui LI ; Yong-Zhen ZHANG
Chinese Journal of Epidemiology 2010;31(9):1030-1034
Objective Genetic analysis was performed to infer the relationship between hantaviruses carried by Rattus norvegicus from Henan and Neimenggu provinces and the other known hantavirus and the vaccine strain. Methods Total RNA was extracted from lung tissues with Trizol reagent. The complete M and S segment sequences of strains NM133 and Q12 were amplified by RT-PCR. The purified DNA fragments were directly subjected to sequencing, and then to sequence analysis and phylogenetic analysis. Results The complete S segment sequences of strains NM133 and Q12 were found to be 1770 nt and 1772 nt in length respectively, with one open reading frame encoding 429 amino acids. The complete M segment sequences of both two strains are 3654 nucleotide in length encoding a protein of 1133 amino acids. The two strains shared a high degree of homology with most of known Seoul virus (SEOV) but quite different from Hantaan virus and other hantaviruses. Furthermore, the nucleoprotein and glycoprotein of the two strains had the congruent structure with the vaccine strain Z37. On the S- and M-phylogenetic trees, both strains (NM133 and Q12) were grouped into the first cluster of SEOV, and were more closely related to the strains, such as: Hb8610, R22, HB55, L99, and K24-e7. Conclusion Both strains (NM133 and Q12) belonged to SEOV, and sharing a high degree of homology and similar secondary structure with strains including the vaccine strains Z37, our data suggested that the present vaccine used in China could effectively prevent HFRS caused by SEOV.