1.Optimization of Fermentation Conditions of Bacillus mucilaginosus Mutant 021120
Xiu-Fang HU ; Fei-Xiang YING ; Ji-Shuang CHEN ;
China Biotechnology 2006;0(09):-
Fermentation conditions for the production of spores of Bacillus mucilaginosus mutant 021120 were optimized through the single factor and orthogonal experiments.The results showed that the biomass and formation of spores were obviously affected by carbonate,followed by nitrogen.Addition of CaCO3 and enhancing air flux notably promoted the formation of spores.The optimal culture medium was composed of 2% starch,0.4% Yeast,0.1% K2HPO4,0.1% MgSO4?7H20 and 0.5 %CaCO3,pH7.5.6% of the inoculum prepared with two-stage extensive culture was inoculated into a 70L fermentor.The fermentation was carried out with 2.0~2.5 vvm of air flux at 32℃ for 38~42h,and the spores of 9.80?108 cfu ml-1 was obtained.Under these optimal fermentation conditions,the capsule was successfully controled and the formation of the spores was effectively improved,thus the satisfying bacteial product was obtained.
2.Prokaryotic Expression,Purification and DNA Binding Activity of DEK Protein's Carboxy-terminal DNA-binding Region
Ying HUA ; Hong-Gang HU ; Xiang-Lei PENG ;
China Biotechnology 2006;0(08):-
DEK protein's carboxy-terminal DNA-binding region(CBD)is a newly found DNA-binding domain of DEK,which contains several phosphorylation sites and has a close correlation with DEK protein's function in vivo and in vitro.Using prokaryotic expression system,the peptide of DEK protein's carboxy-terminal DNA-binding region(CDB)was expressed and purified.In detail,the CDB DNA fragment was constructed into pET-30a(+)vector,and E.coli BL21(DE3)competent cells were used as host cells.The fusion protein His-CBD was expressed by induction of IPTG and purified by Ni-NTA agarose.The result of SDS-PAGE showed that the molecular weight of the purified protein was about 10.7kDa.Electrophoretic mobility shift assay(EMSA)indicated that DEK-CDB prefered to bind to supercoiled form of DNA in vitro,it had similar character to the binding of whole length DEK protein with DNA.This suggested that the carboxy-terminal DNA-binding region of DEK protein might function on the binding of DEK protein to DNA partly.
4.Investigation on occupational norma hexane poisoning accident in population.
Zu-ying HU ; Jian-yong CHEN ; Ning-xiang ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(7):447-447
Accidents, Occupational
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Adolescent
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Adult
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Female
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Hexanes
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poisoning
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Humans
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Male
5.Clinical Research on Treatment of Rheumatoid Arthritis of Cold-dampness Syndromes by Combination of Wen-Jing San-Han Tong-Luo Decoction and Acupuncture
Ping ZHOU ; Ouna CHEN ; Yongmei SU ; Yanghong XIANG ; Yumei WU ; Hong HU ; Ying ZHOU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):784-789
This study was aimed to explore clinical effect on treatment of rheumatoid arthritis (RA) of cold-dampness syndrome by the combination of Wen-Jing San-Han Tong-Luo (WJSHTL) decoction and acupuncture. A total of 88 RA patients were selected and randomly divided into the control group and the treatment group. All 44 cases in the control group were orally administrated with diclofenac sodium, sulphasalazine (SASP) and methotrexate (MTX). All 44 cases in the treatment group were treated with the combination of WJSHTL decoction and acupuncture on the ba-sis of western medication the same as the control group. Each group was treated for three months. Improvements of clinical symptoms, body signs and laboratory indexes were observed. The results showed that clinical symptoms, body signs and laboratory indexes were greatly improved in the comparison before and after treatment in both groups with statistical significance (P < 0.05). Compared to the control group, the treatment group had no statistical significance in the improvement of rest pain, swelling numbers and index, tenderness numbers and index, morning stiffness time, liver and kidney function. Compared with the control group, the treatment group had more obvious advantages in the improvement of joint function, hand grip strength, erythrocyte sedimentation rate (ESR) decreasing, C-reactive protein (CRP) and rheumatoid factor (P< 0.05). It was concluded that compared with single western medication, the combi-nation of WJSHTL decoction and acupuncture with western medication had certain advantages in the improvement of joint function of RA and management of patient's disease condition.
6.Basic fibroblast growth factor-induced differences in gene expression of human umbilical cord blood CD34~+ and CD133~+ stem cells: Gene chip analysis
Huiping CHEN ; Qianru LI ; Jing ZHANG ; Ying DU ; Bo YANG ; Guoxi LI ; Xiang HU ; Ziming DONG
Chinese Journal of Tissue Engineering Research 2010;14(1):75-81
BACKGROUND: Further studies are needed to understand the cytobiological character, functional regulation, gene changes and expression difference of CD34~+ and CD133~+ stem cells induced by basic fibroblast growth factor (bFGF) using gene chip. OBJECTIVE: To compare the differences of gene expression and the response to bFGF of human umbilical cord CD34~+ and CD133~+ cells, and to explore gene expression changes of bFGF-induced umbilical cord CD34~+ and CD133~+ hematopotic stem cells/hemapoietic progenitor cells in vitro. METHODS: Human umbilical cord blood CD34~+ and CD133~+ cells were isolated and purified by MiniMACS immunomagnetic beads selection. The CD34~+ and CD133~+cells were cultured for 10 to 15 days in DMEM/F12 medium, supplemented with bFGF and B27. Total RNA from these cells was extracted and the genetic level of these cells was performed using Oligo GEArray(r) chip and GEArray software. Selected rate of CD34~+ and CD133~+ hematopoietic stem cells was detected using flow cytometry. CD34~+ and CD133~+ cell morphological changes were measured before and after bFGF induction. The concentration and purity of RNA were determined by agarose gel electrophoresis degeneration. Gene-chip test results were analyzed.
7.Effects of combined human thymus and cord blood hematopoietic stem/progenitor cell transplantation on cellular immunity of nude mice
Qingjie ZHAO ; Qianru LI ; Ying DU ; Qingyong ZHANG ; Guoxi LI ; Xiang HU
Chinese Journal of Tissue Engineering Research 2011;15(23):4199-4202
BACKGROUND: T cells are believed to play an important role in anti-infection, anti-tumor and immune function. However, the mechanism underlying the differentiation and development remains poorly understood. OBJECTIVE: To investigate the distribution of T cells in nude mice that are jointly transplanted human thymus and cord blood and the reconstruction of the immune function. METHODS: Thirty Balb/c nu/nu nude mice were randomly divided into two groups: an experimental group and a control group. In the experimental group, human thymus tissue was transplanted into the renal capsule of nude mice. Two weeks later, freshly isolated human cord blood CD34+ cells suspension was back perfused into the nude mice via the vein. In the control group, CD34+ cells transplantation was performed directly without thymus transplantation. After 60 days of breeding, the immune function of nude mice was detected in two groups. RESULTS AND CONCLUSION: Human thymus tissue in the renal capsule of nude mice survived and expressed CD3 and HLA-DR molecule. In the experimental group, CD3+ cells which distributed in the form of dots were observed in the mouse spleen. The proportion of CD3+, CD4+, CD8+, and CD4+CD25+ cells were significantly higher in the experimental group than in the control group. The nude mice from the experimental group rejected human gastric cancer BGC823 cells, while those from the control group did not. These findings demonstrated that combined human thymus and CD34+ cell transplantation allow nude mice to acquire T cell-mediated cellular immune function and possess the ability of anti-tumor.
8.Role of glucagon-like peptide-1 analogue liraglutide played in the proliferation of CD4+ CD25-T cells in normal people and type 1 diabetic patients in vitro
Ying HU ; Xin SU ; Lingjia LIU ; Yufei XIANG ; Qiqi YU ; Shounan YI ; Zhiguang ZHOU
Chinese Journal of Endocrinology and Metabolism 2013;(6):474-478
Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25 T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the therapy of type 1 diabetes.Methods CD4+ CD25-T cells of 10 normal people and 10 newly-onset type 1 diabetic patients were separated from peripheral blood by MACS immunomagnetic beads and stimulated by Human T-Activator CD3/CD28 Dynabeads to proliferate.CFSE labeling technique was used to evaluate the proliferation of CD4+ CD25-T cells by flow cytometry.Liraglutide of different concentrations(0,25,50,and 100 nmol/ml) was added to the proliferation system,then the proliferation of CD4+CD25-T cell was measured.Results (1) Liraglutide suppressed the proliferation of CD4+ CD25-T cells from either normal people or type 1 diahetic patients with dose-dependent manner (P < 0.05).(2) Under the different concentrationsofliraglutide,the proliferation ofCD4+CD25 T cells from diabetic patients was mueh more robust than that of normal people (P<0.01).(3) The inhibitory effects of liraglutide on CD4+ CD25-T cells proliferation in normal people and diabetic patients were similar (P>0.05).Conclusion The proliferation of CD4+ CD25 T cells in type 1 diabetic patients was more robust than normal people,which indicated cellular immune dysfunction in type 1diabetes.Liraglutide inhibits the proliferation of CD4+ CD25-T cells of type 1 diabetic patients in vitro.The immunosuppression effect of liraglutide may have potential value in the treatment of type 1 diabetes.
9.The impact of obstructive sleep apnea-hypopnea syndrome on early renal damage among patients with type 2 diabetes mellitus
Yang WU ; Xiaohong JIANG ; Fei HUA ; Ying TANG ; Long WANG ; Shoukui XIANG ; Jinfeng HU ; Kun WANG
Chinese Journal of Endocrinology and Metabolism 2016;32(3):217-219
[Summary] The aim of this study was to examine the association between obstructive sleep apnea-hypopnea syndrome ( OSAHS ) and microalbuminuria in type 2 diabetes mellitus patients. We found that severe OSAHS significantly increases the risk of early renal damage in type 2 diabetes mellitus patients with HbA1C<7% ( lowest oxygen saturation:OR=2. 41, 95% CI 1. 19-8. 08; apnea hyponea index: OR=2. 91, 95% CI 1. 50-9. 11), suggesting that OSAHS may increase the risk for early renal damage in type 2 diabetes mellitus, especially in those with successful control of glucose.
10.High-frequency oscillatory ventilation in children with hematologic neoplasms and other causes induced acute hypoxic respiratory failure
Long XIANG ; Jian ZHANG ; Hong REN ; Juan QIAN ; Biru LI ; Ying WANG ; Xiaowei HU
Chinese Pediatric Emergency Medicine 2014;21(8):508-512,516
Objective To evaluate the significance of high-frequency oscillatory ventilation(HFOV) used in acute hypoxic respiratory failure(AHRF) children,failing to conventional ventilation.Methods This was a retrospective study of AHRF children ventilated by HFOV from January 2011 to September,2013.All patients were initially treated by conventional mechanical ventilation (CMV),and changed to be treated by HFOV if the patient met to one of the following criteria after the CMV parameters of PIP > 30 mmH2O(1cmH2O =0.098 kPa) or PEEP > 10 cmH2O with FiO2 100% ∶ (1) SpO2 < 90% or PaO2 < 60 mmHg (1 mmHg =0.133 kPa) ; (2) severe respiratory acidosis (PaCO2 > 80 mmHg) ; (3) serious air leakage (mediastinal emphysema or pneumothorax).The following parameters were recorded:patient's gender,age,living PICU time,CMV ventilation time,HFOV ventilation time.We reviewed ventilation parameter settings (MAP,△P,F,FiO2),oxygenation index(PaO2/FiO2,OI),arterial blood gas,heart rate,blood pressure at different time points including late CMV(H0),2 h after HFOV(H2),6 h after HFOV(H6),12 h after HFOV(H12),24 h after HFOV (H24) and 48 h after HFOV (H48),respectively.Various indexes at different time points were compared between survival group and death group,oncology group and no-oncology group.Results PaO2 at H2 compared with H0 had significant improvement[76.9(61.9 ~ 128.0) mmHg vs 50.1 (49.5 ~68.0) mmHg,P =0.006] . PaO2/FiO2 at H2,H48 had significant improvement compared with those at H0,H24 [94.9(66.8 ~ 138.9) mmHg vs 68.0(49.5 ~86.8) mmHg,P=0.039; 135.0(77.6~240.0) mmHg vs 90.7 (54.6 ~161.7) mmHg,P =0.023)].All children's systolic pressure,diastolic blood pressure,heart rate at various time points had no difference (P >0.05).Compared to death group(n =14),PaO2/FiO2,OI at H6,H12,H24,H48 in survival group (n =9) had significant improvement(P < 0.05).Compared to oncology group (n =10),OI at H2,H6 in no-oncology group(n =10) had significant improvement [(19.2 (13.9 ~ 26.6) vs 33.8 (19.7 ~ 48.3),P =0.049 ; 16.0(8.4 ~27.1) vs 28.9(20.9 ~38.9),P =0.027)],and mean airway pressure between two groups at H2,H6,H12 had significant improvement(P < 0.05).Mortality had no significant differcence between two groups (4/10 vs 10/13,P =0.086).Conelusion HFOV used in children with AHRF which had failed with CMV ventilation can improve the patient's PaO2 and OI.Heart rate and blood pressure are stable during HFOV treatment.Oncology group patients needed higher initial MAP to improve oxygenation than no-oncology group patients when changed to HFOV treatment,but the mortality showed no difference between two groups.