1.Lipid synthesis by an acidic acid tolerant Rhodotorula glutinis.
Zhangnan LIN ; Hongjuan LIU ; Jian'an ZHANG ; Gehua WANG
Chinese Journal of Biotechnology 2016;32(3):339-346
Acetic acid, as a main by-product generated in the pretreatment process of lignocellulose hydrolysis, significantly affects cell growth and lipid synthesis of oleaginous microorganisms. Therefore, we studied the tolerance of Rhodotorula glutinis to acetic acid and its lipid synthesis from substrate containing acetic acid. In the mixed sugar medium containing 6 g/L glucose and 44 g/L xylose, and supplemented with acetic acid, the cell growth was not:inhibited when the acetic acid concentration was below 10 g/L. Compared with the control, the biomass, lipid concentration and lipid content of R. glutinis increased 21.5%, 171% and 122% respectively when acetic acid concentration was 10 g/L. Furthermore, R. glutinis could accumulate lipid with acetate as the sole carbon source. Lipid concentration and lipid yield reached 3.20 g/L and 13% respectively with the initial acetic acid concentration of 25 g/L. The lipid composition was analyzed by gas chromatograph. The main composition of lipid produced with acetic acid was palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid, including 40.9% saturated fatty acids and 59.1% unsaturated fatty acids. The lipid composition was similar to that of plant oil, indicating that lipid from oleaginous yeast R. glutinis had potential as the feedstock of biodiesel production. These results demonstrated that a certain concentration of acetic acid need not to be removed in the detoxification process when using lignocelluloses hydrolysate to produce microbial lipid by R. glutinis.
Acetic Acid
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Biofuels
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Biomass
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Culture Media
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Fatty Acids
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Hydrolysis
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Industrial Microbiology
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Lignin
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chemistry
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Linoleic Acid
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Lipids
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biosynthesis
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Oleic Acid
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Rhodotorula
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metabolism
2.Aeroallergen Specific IgE in Serum of Patients with Allergic Rhinitis
Qintai YANG ; Xuekun HUANG ; Yulian CHEN ; Gehua ZHANG ; Yuan LI
Journal of Sun Yat-sen University(Medical Sciences) 2009;30(4):446-449
[Objective] To study the aeroallergens in the patients with allergic rhinitis in Guangzhou.[Methods] Using the German screen allergens quantitative detection system to determinate serum aeroallergen specific IgE and total IgE levels of 625 cases with allergic rhinitis diagnosed in The Third Hospital of SUN Yat-sen University.The samples were stratified on two age groups:the juvenile and the adult.Difference of serum aeroallergen specific IgE of the two groups was investigated.[Results]Aeroallergen specific IgE antibody in the serum of 625 patients with allergic rhinitis were positive.Total IgE in serum was positive in 413 cases,accounting for 66.1%.The positive rate of total IgE was less than that of aeroallergen specific IgE.The positive rate of aeroallergen specific IgE in order were mite mixture (84.32%),cockroach (19.04%),cypress (15.20%),cat-dog epithelia (13.12%),tree mixture (7.20%),ragweed (5.12%),humulus lupulus (1.76%),mugwort (1.76%),mould mixture (1.44%).In the two groups,the positive rate of aeroallergen specific IgE in order were similar.There were significantly higher sensitivity and positive rate of mite mixture in the juvenile group than the adult group (P < 0.01).[Conclusion] The most important aeroallergens are mite mixture,cockroach,cypress,cat-dog epithelia,which could be referenced as Guangzhou patients .The juvenile is more sensitive to mite than the adult.
3.Compliance to sublingual immunotherapy in patients with allergic rhinitis in Guangzhou city
Xuekun HUANG ; Xifu WU ; Qintai YANG ; Yulian CHEN ; Gehua ZHANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2015;(12):616-618
ABSTRACT]OBJECTIVETo study the compliance to sublingual immunotherapy(SLIT) in patients with allergic rhinitis(AR) in Guangzhou city.METHODSFrom January 2014 to May 2014, 202 patients with AR received SLIT were followed up by telephone. According to age, the patients were divided into group A(age<14 years) and group B(age≥14 years). The compliance to SLIT was analyzed and the reasons of poor compliance were investigated.RESULTSAmong 202 patients, only 93 cases(46.04%) were successfully followed up by telephone, 109 cases(53.96%) were lost to visit. Among the 93 cases of successful follow-up, the good compliance rate was 29.03%(n=27), the poor compliance rate was 70.97%(n=66). compliance to SLIT was not affected by age and gender(P>0.05). Main reasons for poor compliance included poor efficacy (48.48%), insufficient education about SLIT (16.67%), inconvenience (15.15%), and adverse reactions(10.61%).CONCLUSION In Guangzhou city, lost follow-up rate in AR patients receiving SLIT is high. Compliance to SLIT is relatively low and improvements shall be made.
4.Expression of Th1、Th2-typed cytokines and its significance in nasal polyps
Huabin LI ; Geng XU ; Yuan LI ; Mingqiang XIE ; Gehua ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2001;(2):51-52
Objective:To evaluate the possible role of cytokines in pathophysiology and treatment of nasal polyps.Method:The expressions of Th1-typed cytokines IFN-γ、IL-2 、IL-12 and Th2-typed cytokines IL-4、IL-5、IL-8、IL-10、IL-13 were investigated with enzyme-linked immune sorbent assay(ELISA) in 25 patients with nasal polyps.Result:There was a significant upregulation of Th2-typed cytokines IL-4、IL-5、IL-8、IL-10、IL-13 in nasal polyps compared with normal nasal mucosa, especially IL-4 and IL-5 (3.9 times and 8.8 times higher than normal mucous respectively) while the expression of Th1-typed cytokines IFN-γ、IL-2、IL-12 drcreased after treated with local glucocorticoid. The levels of Th2-typed cytokines decreased significantly and Th1-typed cytokines had no obvious change.Conclusion:The upregulation of Th2-typed cytokines such as IL-4、IL-5、IL-8、IL-10、IL-12 may play an important role in the pathophysiology of nasal polyps and Th2-typed cytokines can be viewed as a target of treatment to nasal polyps.
5.Improvement on primary culture of human nasal epithelium by enzymatical dissociation
Jihong YANG ; Gehua ZHANG ; Yan WEI ; Yuan LI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;(23):1066-1068
Objective:To highlight the key points of primary culture of human nasal epithelial cells by enzymatical dissociation for high achievement ratio,and to establish a successful primary culture model for subsequent experiments.Method:Primary culture of human nasal epithelial cells was performed with enzymatical dissociation of isolated tissue in serum-free medium. On the basis of this method,some improvements were subjected,such as stripping mucosal epithelium from adjacent connective tissue,applying DNase type Ⅰ to digesting procedure,adding trypsin directly to the collagenase solution containing digested mucosa pieces,employing uncoated culture dishes and so on. Immunofluorescence with a monoclonal anti-cytokeratin antibody 8/18 was used to confirm the epithelial nature of the cultured cells. Result:Nasal epithelial cells grew well and confluenced on the 6th to 8th day. Positive expression of cytokeratin(CK)8/18 showed the epithelial property of cultured cells.Conclusion:Primary culture model of human nasal epithelial cells can be successfully established by enzymatical dissociation. Improvements on processes of material using and enzyme digestion can gain a high achievement ratio and harvest a high purity and certain amount of reliable primary epithelial cells.
6.Differential proteomic analysis of fluconazole-sensitive Candida albicans before and after tetrandrine treatment
Laiqiang GAO ; Hong ZHANG ; Xiaoli ZHANG ; Hui GUO ; Zhiyuan WANG ; Xifu WU ; Gehua ZHANG
Chinese Journal of Dermatology 2011;44(8):560-563
Objective To study the effects of tetrandrine on the protein expression profile of C. albicans,and to screen for proteins associated with the effect of tetrandrine. Methods Fluconazole-sensitive C. albicans CA-3 was cultured with or without tetrandrine (250 mg/L) for 6 hours followed by the collection of Candida cells. Total proteins were extracted from these cells and separated by using immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis. Protein spots were detected and analyzed by Image Master 2D Platinum software, and MALDI-TOF/TOF-MS-based method was used to identify these proteins. Results The two-dimensional gel electrophoresis maps of C. albicans proteins before and after the treatment with tetrandrine were successfully obtained with high repeatability. Image analysis revealed a total of 26 differentially expressed protein spots in C. albicans treated with tetrandrine, and 7 differentially expressed proteins were identified by the mass chromatographic analysis, including 1 up-regulated protein (Pst1), and 6 down-regulated proteins (Idh1,Asc1, Rps5, Asn1, Asn1, Srb1 ). Conclusions The expressions of some proteins in fluconazole-sensitive C. albicans experience significant changes after tetrandrine treatment, and Pst1, Idh1, Asc1, Rps5, Asn1, Asn1 and Srb1 may considerab1y contribute to the effects of tetrandrine on C. albicans.
7.Proteomic analysis of plasma membrane from fluconazole-resistant Candida albicans
Xiaoli ZHANG ; Laiqiang GAO ; Xifu WU ; Hong ZHANG ; Gehua ZHANG ; Xingfeng YIN
Chinese Journal of Dermatology 2013;(4):262-265
Objective To identify and analyze plasma membrane proteins differentially expressed between fluconazole-sensitive and-resistant C.albicans strains.Methods Two C.albicans strains from a same parent,including the fluconazole-sensitive C.albicans strain CA-3 and fluconazole-resistant C.albicans strain CA-16,served as the subject of this study.Plasma membrane proteins were isolated from both of the C.albicans strains,and subjected to two-dimensional polyacrylamide gel electrophoresis analysis for the screening of differentially expressed proteins,which were then identified by using matrix assisted laser desorption/ionization time-of-flight mass spectrometry.The resultant data were searched against a protein database for C.albicans.Results Twentytwo proteins were identified to be differentially expressed between the fiuconazole-resistant and-sensitive C.albicans strain.Of them,6 proteins (Adh1p,Csp37p,Pgk1p,Pgk1p and 2 unnamed proteins,i.e.,gi227305312and gi53954641) were highly expressed,while 16 proteins (Aco1p,Aco1p,Hsp78p,Gut2p,Sdh12p,Ilv2p,Ndh51p,Ndh51p,Atp1p,Pda1p,Srb1p,Idh1p,Tdh1p,Cyt1p,Cox4p,Cox13p) were lowly expressed in the fluconazole-resistant C.albicans strain compared with the fluconazole-sensitive strain.Conclusion The plasma membrane proteins differentially expressed between fluconazole-sensitive and-resistant C.albicans strain are mainly implicated in energy metabolism and mitochondrial function.
8.Preparation of a functional monoclonal antibody against human 4-1BBL molecule and analysis of its biological characteristics
Xudong WANG ; Bin ZHOU ; Wenxiang LI ; Gehua YU ; Guangbo ZHANG ; Xueguang ZHANG
Chinese Journal of Immunology 1985;0(05):-
Objective:To prepare functional monoclonal antibodies against human 4-1BBL molecule and analysis of their biological characteristics.Methods:Female BALB/c mice of 6-8 weeks old were immunized with 4-1BBL transfectant (L929/4-1BBL) as immunogen.The spleen B cells of the mice were fused with sp2/0 and hybridoma cells were screened with 4-1BBL transfectant (L929/4-1BBL) by FCM.The biological characteristics of antibody were investigated by rapid isotyping analysis,karyotype analysis,competitive inhibition test etc.Furthermore,the growth of monocytes in vitro was determined by cell number counting and the cytokine concentration in the supernatants was assayed by ELISA.Results:One hybridoma cell line named 3E7 was obtained,which had the property of secreting anti-human 4-1BBL monoclonal antibody continuously and steadily.This mAb specifically recognized human 4-1BBL molecules.The experimental results manifested that mAb 3E7 could effectively enhance the growth of monocytes and the high level IL-6 and TNF-? secretion.Conclusion:One hybridoma cell line which can secret a functional mouse anti-human 4-1BBL mAb has been developed successfully.This mAb can specifically recognize human 4-1BBL and regulate growth and functions of monocytes in vitro.
9.Clinical study on postoperative analgesia following sinonasal surgery
Xiaowen ZHANG ; Yuan LI ; Gehua ZHANG ; Xian LIU ; Qintai YANG ; Jiancong HUANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(12):-
0.05), though scores of both group were significantly lower than that of group T(P
10.Correction between cytokine expressions in circulating CD4~+ CD28~-T cells and Graves'ophthalmopathy
Zhiping SUN ; Weixue ZHONG ; Heng MIAO ; Gehua YU ; Na YOU ; Xueguang ZHANG
Chinese Journal of Endocrinology and Metabolism 2010;26(1):39-40
The immune effect of CD4~+CD28~-T cells on Graves'ophthalmopathy(GO)was investigated.The expressions of interferon-γ(IFN-γ),interleukin-2(IL-2),and IL-4 in CD4~+ CD28~-T ceils were assayed by flow cytometry in GO patients,Graves'disease(GD)patients without ophthalmopathy,and healthy control subjects.The results showed that the percentage of CD4~+CD28~-T cells significantly increased in GO patients(P<0.05),with increased IFN-γ expression(P<0.05)and decreased IL-2 expression(P<0.05).These changes were closely correlated with clinical activity score(P<0.05).There were no significant differences in IL-4 expression among three groups.The resuh suggests that CIM~+ CD28~- T cells which hishly secrete IFN-γare related to the pathological lesion of GO.