3.Effects of hypoxia on the capsule size of Cryptococcus neoformans
Xue DU ; Qingtao KONG ; Suyang HUANG ; Rui YANG ; Hong SANG
Journal of Medical Postgraduates 2015;(8):799-803
Objective Little is known about the role of low oxygen concentration in the formation of the capsule of Cryptococ -cus neoformans .This study was to investigate the effect of hypoxic condition on the capsule size of Cryptococcus neoformans . Methods We cultured 39 Cryptococcus neoformans strains in liquid Sabouraud medium and then incubated them at the normoxic (21%) and hypoxic concentration (1%), respectively.After 72 hours of incubation, we observed the capsules formed at different ox-ygen conditions under the Olympus microscope and the measured their sizes using the Axiovision software . Results Of the total number of Cryptococcus neoformans strains , 30 showed larger capsules in the normoxic than in the hypoxic condition , 17 with statisti-cally significant differences in their diameters (P<0.05), while the other 9 strains exhibited just the opposite results , 4 with statisti-cally significant differences in their diameters ( P<0.05). Conclusion Hypoxia might inhibit the growth of the Cryptococcus neoformans capsule.However, further studies are needed on its correlation with the serum or gene types of the Cryptococcus neofor-mans strain.
4.Effects of bencycloquidium bromide on the expression of MUC5 AC induced by lipopolysaccharide in cultured human nasal epithelial cells
Min YANG ; Xue LU ; Jiangju HUANG ; Rui LONG ; Juan LI
Chinese Pharmacological Bulletin 2016;32(6):783-788
Aim Toinvestigatetheeffectofbencyclo-quidium bromide(BCQB)on mucus MUC5AC expres-sion induced by lipopolysaccharide in cultured human nasalepithelialcells(HNECs).Methods Primary culture of human nasal epithelial cells (HNECs)was randomly divided into control group (C,with no treat-ment),LPS group (LPS,with LPS 1 mg · L-1 added in),BCQB low dose group(BCQBL,with LPS 1 mg· L-1 and BCQB 10 -8 mol·L-1 added in),BCQB mid-dle dose group(BCQBM,with LPS 1 mg·L-1 and BC-QB 10 -7 mol·L-1 added in),BCQB high dose group (BCQBH,with LPS 1 mg·L-1 and BCQB 10 -6 mol· L-1 added in)and ipratropium bromide group(IB,with LPS 1 mg·L-1 and IB 10 -6 mol·L-1 added in).Af-ter incubation at 37 ℃with 5% CO2 for 24 h,the ex-pression of MUC5 AC mRNA was detected with Real-time PCR and the expression of MUC5 AC protein in HNECs was detected with Western blot,while the ex-pression of MUC5 AC protein in supernatant was detec-tedwithELISAineachgroup.Results Ascompared with control group,the expression of MUC5 AC mRNA and protein increased significantly in LPS group (each P<0. 01 ).As compared with LPS group,the expres-sion of MUC5 AC mRNA and protein decreased signifi-cantly in each group of BCQB(P<0. 01,P<0. 05), and there was no statistical difference between BCQB high dose group and control group (each P>0. 05 ). Conclusion Bencycloquidiumbromidecansuppress MUC5 AC expression induced by LPS in cultured hu-man nasal epithelial cells,indicating that BCQB may be a new drug for nasal mucous hypersecretion diseases.
5.Development of Novel Microbial Lipase Resources
Zheng-Yu SHU ; Long-Yin XUE ; Rui-Feng LIN ; Shao-Li CAI ; Jian-Zhong HUANG ;
Microbiology 1992;0(05):-
Microbial lipase,one of important industrial biocatalysts,has been used widely in many industrial and agricultural fields.It is always the research focus to screen,mine and develop the microbial lipases with novel catalytic activity and high stability.This paper introduces briefly the pathways and methods to mine novel microbial lipase resources from six aspects,including extremophile,metagenome,genome database,protein engineering,immobilization,chemical modification,etc.
6.Protein Engineering of Microbial Lipases
Rui-Feng LIN ; Zheng-Yu SHU ; Long-Yin XUE ; Huan JIANG ; Jian-Zhong HUANG ;
China Biotechnology 2006;0(09):-
Microbial lipases are important industrial biocatalysts with the character of stereoselectivity,site selectivity and high catalytic activity with few side effects.They have been used widely in many industrial and agricultural fields.The technology of protein engineering has been successfully applied to improve the activity and stability of microbial lipases,which will raise the competitive capacity of microbial lipase preparations and enlarge theirs application fields.The strategies,the problems and the prospects of protein engineering technology which have been applied to modify the microbial lipases was surveied.
7.Polyclonal antibody specific for hypervariable region of VP1 capsid protein of Taura syndrome virus (TSV) prepartion and characteristic analysis
Xinxin HUANG ; Chengang YUAN ; Xue NING ; Ming GU ; Qiang CAI ; Rui LIU ; Chengping LU
Chinese Journal of Immunology 2015;(2):221-224,229
Objective:Preparation and immune characteristic analysis of polyclonal antibody against hypervariable region protein of Taura syndrome virus major capsid protein VP 1 as a reference for studies on immunological diagnosis reagent.Methods:The recombinant vector pET-VP1 was transformed into E.coli BL21 for protein expression.Immunizing a New Zealand rabbit with purified VP1 protein,the titer of anti-VP1 serum was determined by Agar diffusion test and ELISA.Monoclonal phage specific binding to the purified VP1 protein was used for competitive inhibition test.Results: The VP1 protein was soluble and high expression in E.coli BL21.The biological activity titer of anti-VP1 serum reached 1∶26 ,1∶217 determined by Agar diffusion test and ELISA respectively.A litter binding activity of antiserum and VP 1 protein could be blocked by monoclonal phage , but would not affect the final positive result.Conclusion:High titer antibody Preparation of the VP 1 hypervariable region protein.The binding activity of the polyclonal antibody with VP1 protein was not affected by the mutations of VP 1 protein in minority areas ,so the antiserum could be used as immu-nological detection diagnosis agent.
8.Study on Interaction Mechanism between 3 Kinds of Flavonoids Compounds and Human Serum Albumin by Fluorescence Spectrometry
Rui LAN ; Xiaobao GONG ; Ligua HUANG ; Zhu CHEN ; Xue ZENG ; Baoshun ZHANG
China Pharmacy 2016;27(22):3054-3057
OBJECTIVE:To study the interaction mechanism between flavonoids and human serum albumin (HSA),and to compare the effects of different B-ring substitutions(hydroxyl,methoxyl group)of flavonoids on macromolecular receptor. METH-ODS:The interaction regularity between three flavonoids with different B-ring substitutions(quercetin,hesperetin,methyl hespere-tin) and HSA was studied with fluorescence spectroscopy,the fluorescence quenching types between 3 flavonoids and HSA were determined and analyzed,and the velated binding constant,binding site and thermodynamic parameters were calculated. RE-SULTS:The quenching constants (Ksv) and binding constants (KA) were decreased with the increase of temperatures. The number of binding site(n)was approximately equal to one,and the thermodynamic parameters ΔH<0,ΔS>0,the binding interaction of these compounds with macromolecules was influenced because of the difference of the B-ring substituents. CONCLUSIONS:The quenching mechanism between three flavonoids and HSA was static quenching;the number of binding site was one;the interaction force of the three compounds with HSA was mainly static electricity,and hydroxyl group in the B-ring was likely the major active group and exerted stronger binding force than methoxyl group to connect with macromolecules.
9.Establishment of a model of granuloma infected by Trichophyton rubrum in BalB/C mice
Suyang HUANG ; Qingtao KONG ; Yongnian SHEN ; Xue DU ; Rui YANG ; Hong SANG ; Weida LIU
Journal of Medical Postgraduates 2016;29(5):485-489
Objective Trichophyton rubrum strains can cause superficial infection and also deep tissue infection, but relevant animal model has not been reported yet.The aim of this study was to construct an animal model of granuloma infected by T.rubrum. Methods Three T.rubrum strains isolated from clinical granuloma tissues, 2 T.rubrum strains isolated from tinea infection and a standard strain of ATCCMYA4438 were selected.Corticosteroids were given to the Balb/C mice before and after the injection of the T. rubrum and mucin suspension and the mice model of granuloma was established.Direct microcopy, culture and histopathologic method were adopt to verify the infection effects. Results The mice inoculated with the T.rubrum granuloma strains with mucin suspension were examined after 21 days in the condition of applying appropriate dose of glucocorticoids.Direct microscopic examination showed the slender mycelium, fungal culture showed the growth of colony and histopathology showed excessive keratinization of foot tissue, formation of granuloma in the dermis with inflammatory cell infiltration of neutro-philic granulocyte and lymphocytes.However, the mice inoculated with the T.Rubrum tinea strains with mucin suspension showed the negative result. Conclusion The rubrum granuloma mice model can be es-tablished using the clinical isolates of T.rubrum granuloma strains with the mucin and glucocorticoids interventions.
10.Clinical features of acute retinal pigment epitheltis
Xue, YAO ; Zhi, LI ; Rui-Juan, WANG ; Jun, MEI ; Lin-Li, WANG ; Xian-Jin, HUANG
International Eye Science 2015;(1):155-158
Abstract?AlM:To analyze of the clinical features of acute retinal pigment epitheltis ( ARPE) .?METHODS: The clinical data of 36 ARPE patients ( 40 eyes) attending this center from January 2008 to January 2014 were reviewed retrospectively. Of them, 21 patients (58.3%) were male (male :female=1:0. 71). The mean age was 40. 92±7. 13 years old (range:17~60y). The mean best-corrected visual acuity (BCVA) was 0. 50±0. 26 with a range of 0. 3 ~ 1. 0. Thirty-two patients were unilateral cases. All the patients were examined for BCVA, funds photography, fluorescein fundus angiography ( FFA ) , optical coherence tomography ( OCT) . FFA was shown as three types: type ▏ to multiple black light or grape variety fluorescent spot; Type II for l lesions visible fluorescence leakage; Type Ⅲ lesions with choroid neovascularization ( CNV ) . OCT was the following three forms: multiple RPE lesions layer reflection intermittent, proliferation ( type ▏); pigment epithelial detachment with limitations neural epithelium ( typeII);types l and ll with CNV ( type Ⅲ) .?RESULTS: Ocular fundus showed that the lesions were multiple dark-gray spots with a dark circumscribed area at the macular or nearby in all 40 eyes. FFA showed:21 eyes were type ▏, 17 eyes were type II and 2 eyes were typeⅢ, BCVA between type ▏ and type II was statistically significant (P<0. 05), the same was between type 芋. BCVA between different cases in the same type and between type II, Ⅲ, was no statistical difference ( P>0. 05). OCT showed 21 eyes wwere type ▏, 17 eyes were type II and type Ⅲ 2 eyes. BCVA average between type▏ andIIwas statistically significant (P<0. 05). The mean BCVA was no statistically significant difference between type II and Ⅲ patients (P>0. 05).?CONCLUSlON:ARPE fundus demonstrated the multiple dark gray discrete lesions, the degree of visual impairment related with the presence of pigment epithelial barrier and lesion location. OCT and FFA characterized three types. FFA is shown asblack light orgrape variety fluorescent spot, and is the basis of diagnosis. OCT can display the lesions organization form of each layer clearly. lt plays a more and more important role in the diagnosis and differential diagnosis of ARPE.