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.Correlation of IL-19 and its receptors with tissue remodeling in chronic rhinosinusitis
Xia LI ; Lihong CHANG ; Zizhen HUANG ; Zhiyuan WANG ; Xianzhen CHEN ; Jun WANG ; Gehua ZHANG
Chinese Journal of Pathophysiology 2017;33(5):919-924
AIM:To explore the differential expression of interleukin-19 (IL-19) and its receptors (IL-20R1/IL-20R2) in the samples of chronic rhinosinusitis (CRS) with or without nasal polyps (CRSwNP and CRSsNP), and to investigate the correlation of IL-19 and its receptors with tissue remodeling factors, including matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in CRS.METHODS:The polyps from CRSwNP patients (n=30), the sinus mucosa from CRSsNP patients (n=15) and the inferior turbinal mucosa of nasal septum de-viation patients (n=15) were collected.The expression of IL-19 and its receptors (IL-20R1/IL-20R2) in each group was detected by real-time RT-PCR and immunohistochemistry.The expression of tissue remodeling factors MMP-2, MMP-9 and TIMP-1 in different groups was detected by real-time RT-PCR and ELISA.RESULTS:Increased mRNA and protein expression levels of IL-19, IL-20R1, IL-20R2 and MMP-9 were observed in CRSwNP group compared with CRSsNP and control group (P<0.05), while elevated expression level of TIMP-1 was observed in CRSsNP group compared with CRSwNP and control group (P<0.05).The relative expression of MMP-2 among the 3 groups showed no obvious difference.The expression of IL-19 and its receptors showed significantly positive correlations with MMP-9 in CRSwNP (P<0.05).No significant correlation between IL-19 and/or its receptors with TIMP-1 in CRSwNP group was observed.CONCLUSION:The high expression levels of IL-19, IL-20R1 and IL-20R2 and their positive correlations with MMP-9 in CRSwNP indica-te that IL-19 and its receptors may be involved in the tissue remodeling of chronic rhinosinusitis.
3.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.
4.Incidence of Otitis Media with Effusion in Adenoid Hypertrophy with Allergic Rhinitis in Children
Huiyi DENG ; Qintai YANG ; Weihao WANG ; Xuekun HUANG ; Jiancong HUANG ; Yulian CHEN ; Peng LI ; Gehua ZHANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):101-105
Objective]In order to study the impact of adenoid hypertrophy(AH)with allergic rhinitis(AR)in the otitis media with effusion(OME)in children,as well as to discuss the risk factors on the inducement of OME in patients with AH.[Methods]The clinical materials were collected and analyzed from 205 children with AH who admitted for surgical treatment between 2013 and 2015, including medical history and signs,acoustic immittance measurement,allergy screen as well as blood routine,and to evaluate the situation of OME and AR in patients with AH. All the data were analyzed by SPSS 20.0.[Results]Among the 205 AH children,66 cases(32.20%)were accompanying with OME. The AH patients aged 3-5 years had the highest incidence of OME,which decreased with age(P=0.018). It is significant on the prevalence of the OME in patients with AH between the two groups with and without AR (P = 0.010). In the logistic model investigating the risk factors of inducing OME among patients with AH,the third-degree AH, Type-three and the accompanying with AR were significant. The third-degree and third-type AH children were 2.729 and 6.390-folds higher than others respectively (P = 0.047,P = 0.001). The incidence of OME appeared to be 1.212-folds higher among patients with AR(P = 0.010).[Conclusion]Mechanical obstruction and AR played a superimposed role in the inducement of OME among patients with AH,which was effected by multiple factors. We should pay attention to the“lateral respiratory”allergic diseases on eustachian tube and middle ear.
5.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.
6.Clinical characteristics in patients with allergic rhinitis and nonallergic rhinitis in Guangzhou
Weihao WANG ; Xuekun HUANG ; Zhuanggui CHEN ; Huiyi DENG ; Yulian CHEN ; Gehua ZHANG ; Qintai YANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2016;23(9):533-536
OBJECTIVE To obtain and compare the clinical characteristics in patients with allergic rhinitis(AR) and nonallergic rhinitis(NAR) and investigate the trends in the proportion of AR and NAR in recent 10 years in Guangzhou. METHODS 5486 patients with nasal hyper-reactivity symptoms were divided into the AR group and NAR group. Clinical data including gender, age distribution and seasonality were analyzed. RESULTS The trends in the proportion of AR and NAR during the past decade did not change significantly. Male made up the majority of AR patients and NAR patients and AR patients were significantly younger than NAR patients. Male AR patients were significantly younger than females, while there were no significant difference in the age distribution among the male NAR patients and female ones. As the age increasing, the proportion of AR and NAR patients in overall study population present opposite tendency. The main onset season in AR was summer and in NAR was winter in Guangzhou city. CONCLUSION There were significant differences between AR and NAR in age, gender and seasonality. However, the trends in the proportion of AR and NAR in recent 10 years did not change significantly in Guangzhou.
7.Distribution and treatment onintractable epistaxis in concealed sites
Qintai YANG ; Huiyi DENG ; Weihao WANG ; Xuekun HUANG ; Lei LV ; Xian LIU ; Gehua ZHANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2016;23(10):602-605
OBJECTIVE To drew the map of distribution of concealed sites in epistaxis and optimize the mode of diagnosis and treatment. METHODS We have searched 117 references on intractable/refractory/latent epistaxis treated under nasal endoscopy in the last decade, including 11 708 cases with epistaxis. There were total 11 860 cases in this study plus 152 cases in our hospital. We summed up the experiences of the optimized treatment mode performed on concealed epistaxis, which was searching the accurate bleeding areas by nasal endoscopy and performing minimally invasive radiofrequency treatment without nasal packing. RESULTS We had found that the offending arteries about epistaxis are mainly sphenopalatine artery and anterior or posterior ethmoid artery. The frequency of bleeding areas came as follows: the vault of inferior nasal meatus in 3783 cases(31.90%), the olfactory sulcus of middle turbinate in 3522 cases(29.70%), the posterior regions of middle meatus in 1349 cases(11.37%), the regions of deviation of nasal septum in 1065 cases(8.98%), the foremost regions of nasal cavity in 738 cases(6.22%), and the others or uncertain in 1403 cases(11.83%). Finally, we drew a concise map of distribution about epistaxis on the basis of the concealed bleeding areas and offending vessels. 151 of 152 patients(99.34%) could find out the bleeding sites accurately and stop the bleeding through minimally invasive radiofrequency ablation. CONCLUSION We drew a map of distribution about epistaxis in concealed areas so that it is convenient for us to seek out the bleeding sites. Rational use of nasal endoscopy to explore the sites of intractable epistaxis and minimally invasive radiofrequency are optimized mode of treatment.
8.Immune intervention effect of human-mouse chimeric antibody B7-1 against murine lupus nephritis model
Hui SHEN ; Yuqiang ZHU ; Yong KONG ; Jing WANG ; Huating ZHU ; Gehua YU ; Lei CAI ; Ying ZHU ; Zhiyao WANG ; Yuhua QIU
Chinese Journal of Immunology 2015;(9):1200-1205
Objective:On the basis of the use of chemical methods to establish mouse model of lupus nephritis and its biological identification , we investigate the reverse effect of pathological lesions of B 7-1 human-mouse chimeric antibody blockade against B7/D28 signaling pathway in mice with lupus nephritis model.Methods:Pristane was injected intraperitoneally to 6-week-old female C57BL/6J mice at dose of 0.5 ml per mouse in one go,and urine protein,ANA and renal pathological changes were detected on a monthly basis.Mice whose urine protein content reached ++and ANA fluorescence intensity reached ++were randomly devided into three groups ,five each.Antibody intervention group was sequentially injected with B 7-1-mouse chimeric antibody by orbital venous , positive control group was injected with immunosuppressant CTX , negative control group was injected with isotype control IgG.Urine protein and ANA were also detected on a monthly basis.Mice were sacrificed three months after intervention was executed.Kidney was used for H&E dying , IC detection and electric microscope observation.Results: After four-month Pristane induction , urine protein content of 80%mice reached +-+++,meanwhile,serum ANA fluorescence intensity reached ++-+++.Glomerulonephritis infiltrating cells were observed Mice with urine protein and ANA , glomerular inflammatory cell infiltration , tubular epithelial cell degeneration visible edema ,vascular congestion significantly ,fibrosis.After antibody intervention ,urine protein content in antibody intervention group gradually reduced from ++-+++to ±-+++,ANA ++-+++to +-++,and were significantly different from that in the negative control group ( P<0.01 ).Analysis of kidney H&E dying showed that antibody glomerular infiltration of inflammatory cells in the intervention group and tubular congestion and other symptoms were improved significantly.Immunofluorescence staining indicated that fluorescence intensity of IC was significantly reduced in the antibody intervention group.Electron dense deposits reduction and glomerular basement membrane uniformity were observed in antibody intervention group by electric microscope when compared with the negative control group.Conclusion:B7-1 antibodies could downregulate immune response through inhibiting B 7-1/CD28 signaling pathway , reducing the production of autoantibodies and reversing pathological damage caused by autoimmune response .
9.Co-stimulatory signal GL50-ICOS expression in thyroid of Graves' disease and its immune pathogenetic significance
Lei CHEN ; Ting LI ; Honger YIN ; Songguang JU ; Gehua YU ; Lian JIANG ; Fengming WANG ; Ming WU ; Xueguang ZHANG
Chinese Journal of Endocrinology and Metabolism 2010;26(11):967-970
Objective To study the expression of co-stimulatory molecules, GL50-ICOS, in thyroid tissue of patients with Graves' disease (GD) and to explore their relationship with the immune pathogenesis of GD.Methods RT-PCR, Western blot, immunohistochemistry were applied to detect the expression of GL50-ICOS in thyroid of GD. Thyrocytes were cultured in the absence or presence of pro-inflammatory cytokines. The expression of GL50 on thyroid follicular cells (TFC) was further measured by flow cytometry. Results (1) In GD patients,the percentage of CD4+ CD28- T cells was significantly increased as compared with the control healthy individuals. The expression of co-stimulatory molecule ICOS was up-regulated. (2) The mRNA level of ICOS was significantly increased in GD patients than that in nontoxic goiter(NTG) patients(P<0.01). (3)Compared with NTG control group, the GL50 protein expression was much higher in thyroid tissues of GD patients (P <0.01). (4)The results of immunohistochemistry showed that GL50 expression was observed in all GD thyroid tissues, while no expression of GL50 was detected in NTG thyroid tissues(P<0. 01). (5) The expression of GL50on primary cultured thyroid follicular cells was significantly increased under the stimulatation of pro-inflammatory cytokines in vitro. Conclusion GL50-ICOS is expressed abnormally in thyroid tissue of patients with GD.
10.Concentration, distribution and expression of interleukin-5 in human nasal polyp tissues.
Rui XU ; Yuan LI ; Minqiang XIE ; Geng XU ; Gehua ZHANG ; Shufang WANG
Chinese Medical Journal 2002;115(9):1386-1389
OBJECTIVESTo study the concentration, distribution and expression of IL-5 in nasal polyp tissues and explore its significance in micro-environment differentiation of eosinophil accumulation.
METHODSThe concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers were used as control.
RESULTSIL-5 concentration in polyp tissues was significantly higher than that in turbinate mucosa (P < 0.05). There was no significant difference in the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentrations in polyp tissues were markedly higher in patients with allergic rhinitis compared with those without (P < 0.05). IL-5 concentrations had no correlation with age and sex (P > 0.05). 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of lymphocytes and neutrophils were positive for IL-5; IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05); there was no significant difference in the the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissue was significantly stronger in patients with allergic rhinitis compared with those without (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and turbinate nasal mucosa (both P > 0.05).
CONCLUSIONIL-5 is the key cytokine in eosinophilic pathologic mechanisms in nasal polyp tissues.
Adult ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; chemistry ; Female ; Humans ; Interleukin-5 ; analysis ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Nasal Mucosa ; chemistry ; Nasal Polyps ; immunology ; Turbinates ; chemistry