2.The effect of umbilical cord mesenchymal stem cells transplantation for the GD mice on regulatory B cells
Rongliang LI ; Koulan HAN ; Xiaoli DAI ; Cheng HUANG ; Weiyong LI
Chongqing Medicine 2015;(9):1168-1170
Objective To investigate the therapeutic mechanism of umbilical cord mesenchymal stem cell(UC-MSCs)trans-plantation for the graves disease(GD)mice .Methods Thirty two mice were divided into 3 groups as following :normal control group (G0) ,GD control group (G1) ,UC-MSCs group(G2) .Enzyme linked immunosorbent assay(ELISA)was used to measure the level of TSAb in blood serum and the expression of FT4 was measured by chemiluminescence .Thyroid sections were stained with hema-toxylin and eosin(HE)for histological examination .Splenocytes were stained with multicolor immunofluorescence and detected by flow cytometry to analyze the percentages of CD1dhiCD5+CD19+ regulatory B cells(Bregs) .Expressions of IL-10 and TGF-βmR-NA in spleen organization were measured by Real-time PCR .Results At 26 weeks ,the level of TSAb in blood serum in G2 was more significantly decreased than in G1(P<0 .05) ,and the level of CD19+ B in spleen in G2 was also more significantly decreased than in G1(P<0 .05) ,however ,the percentage of CD1dhiCD5+CD19+ Bregs splenocytes and the levels of IL-10 and TGF-βmRNA in spleen organization were more significantly increased than in G1(P<0 .05) .The concentration differences of TSAb in serum was negatively correlated with the percentage differences of CD1dhi CD5+ CD19+ Bregs ,however ,positively correlated with the expres-sion differences of IL-10 and TGF-βmRNA in spleen before and after transplantation .Conclusion Activation of Bregs may be one of the mechanisms of UC-MSCs therapeutic effect on GD mice .
3.Transport of geniposide and geniposide in Zhizi Bopi Decoction in MDCK cell membrane model
Jingwen HAN ; Jun LI ; Cheng HUANG ; Zhaolin CHEN ; Tingting HU
Chinese Pharmacological Bulletin 2014;(4):468-472
Aim To study the transport of geniposide and geniposide in Zhizi Bopi Decoction in MDCK cell membrane model. Methods The safety concentration of geniposide and Zhizi Bopi Decoction in MDCK cells were determined by MTT assay. Then the MDCK cell membrane model was used to investigate the transport of drugs. Firstly, the effects of time, drug concentra-tion, P-gp inhibitor and EDTA on the absorption and transport of geniposide were studied systematically. Secondly, the differences were compared between the transport of the same concentration of geniposide as single compound and that in Zhizi Bopi Decoction in MDCK cell model. The drug concentration was deter-mined by high performance liquid chromatography ( HPLC) to calculate the apparent permeability coeffi-cient (Papp). Results Geniposide transport in MDCK cell monolayer was time and concentration dependent. P-gp inhibitors had no significant effect on its transport and the transport of geniposide was enhanced by ED-TA. The absorption Papp of different concentrations of geniposide in Zhizi Bopi Decoction were ( 8. 96 ± 0. 35 ) × 10 -7 cm · s-1 , ( 8. 95 ± 0. 38 ) × 10 -7 cm · s-1 and (9. 16 ± 0. 30) × 10 -7 cm·s-1, significantly higher than the absorption Papp of geniposide as single compound(5. 85 ± 0. 44) × 10 -7 cm·s-1, (6. 88 ± 0. 38) × 10 -7 cm·s-1 and (6. 31 ± 0. 19) × 10 -7 cm ·s-1 ( P<0. 05 ) . Conclusion The transport of ge-niposide in MDCK cell membrane model is passive transport and is not affected by P-gp. Geniposide may transport via the paracellular route. The Zhizi Bopi De-coction can increase the absorption of geniposide.
4.Two family members all elevated blood lead the investigation.
Cheng-hong HAN ; Yun-long DING ; Ling HUANG ; Shi-ping DU ; Lü JI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(9):666-666
Adult
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Child
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Child, Preschool
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Female
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Humans
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Infant
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Lead
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blood
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Lead Poisoning
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blood
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epidemiology
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Middle Aged
5.Expression and Significance of A Novel Gene BC047440 in Multiplicate Malignant Tumor Tissues
Lu ZHENG ; Jing LI ; Ping LIANG ; Tong-Han YANG ; Xiao-Bing HUANG ; Shi-Cheng LIU ;
Chinese Journal of Bases and Clinics in General Surgery 2003;0(03):-
0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P
6.Studies on the Fermentation Conditions and Anti-tumor Effect of Exopolysaccharide from Rhizobium sp.N613
Yong HAN ; Xiao-Bo HUANG ; Yue-Feng DONG ; Hong-Bing CHENG ; Liang-Qi ZHAO ;
Microbiology 1992;0(05):-
The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.
7.Effect of artesunate on OAZ gene expressions in MRL/lpr lupus mice
Rongliang LI ; Koulan HAN ; Cheng HUANG ; Xiaoli DAI ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2014;18(6):380-384,后插1
Objective To investigate the in vivo effects of artesunate (ART) treatment on OAZ gene expressions in MRL/lpr lupus mice.Methods Twenty-four 12-week-old female MRL/lpr lupus mice were randomly divided into two groups by the random number table,i.e.,the lupus control group and the ART treatment group,12 in each group.Besides,another 8 BABL/C mice were recruited as the normal control group.Peripheral blood mononuclear cells (PBMC) were collected from lupus mice before treatment as well as 4 week and 8 week after treatment,and RNA was extracted and reverse transcripted to cDNA.mRNA expression levels of OAZ and Id1-3 were measured by using real-time polymerase chain reaction (PCR),and the data between the two groups were analyzed by t test,a plurality of samples were compared with the single factor analysis of variance.Serum levels of IFN-γ,IL-4,BAFF and ANA were tested by enzyme-linked immunosorbent assay (ELISA).Relationships of the gene expression levels with levels of cytokines and ANA were analyzed.Statistical analysis were uising t test,ANOVA and LSD-t test.Results mRNA expression levels of OAZ,Id1 and Id3 gene (△Ct:12.9±0.8,12.0±0.4,10.2±0.8) in lupus mice were significantly different from 8 weeks after the treatment comparing to those in the lupus control group (△Ct:9.8±1.0,9.3± 1.1,8.1±0.8) and the normal control group (△Ct:13.9±1.2,11.4±0.7,4.7±0.8,10.3±1.0)(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),and alteration of OAZ mRNA expression levels before and after the treatment were positively correlated with changes of Id1,Id3 levels (r=0.867,0.947; P<0.05),and levels of IL-4 and ANA were significantly lower 8 weeks after the treatment than those in the lupus control group (P<0.05); while level of IFN-γwas higher than those in the lupus control group (P<0.05).Alteration of OAZ mRNA expression levels before and after the treatment were negatively correlated with changes of ANA,BAFF levels (r=-0.955,r=-0.937; P<0.05) and positively correlated with changes of Th1/Th2 levels (r=0.976,P<0.01).Conclusion The expression of genes involving in the OAZ and downstream gene are effectively reduced along with the alteration of several cytokines and ANA after ART treatment.OAZ signaling pathway can play an important role in ART treatment for SLE.
8.Isolation and identification of serum exosomes in the patients with polymyositis/dermatomyositis
Aiyan ZHANG ; Cheng CHEN ; Xiaodan WU ; Tianli REN ; Bing GU ; Hongyu HUANG ; Zhijun HAN ; Mingzhu GAO
Chinese Journal of Clinical Laboratory Science 2017;35(8):583-586
Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.
9.Preparation and Determination of Entrapment Efficiency of Puerarin Polymeric Micelles
Xiaoxing MA ; Cuiyan HAN ; Chang LIU ; Dan ZHU ; Xiaoyu SUI ; Cheng YUAN ; Haitao HUANG
China Pharmacy 2016;27(22):3122-3124
OBJECTIVE:To prepare Puerarin polymeric micelles and establish a method to determine its entrapment efficiency. METHODS:Puerarin polymeric micelles were prepared by film dispersion method. The polymeric micelles and free drug were sepa-rated by centrifugal-millipore filter filtration method. The entrapment efficiency of puerarin polymeric micelles was determined by HPLC. Diamonsil C18(2)column was used with 1% citric acid solution-methanol(65∶35)at the flow rate of 1 ml/min. The detec-tion wavelength was set at 250 nm,and column temperature was room temperature. RESULTS:The prepared polymeric micelles were spherical and spherical-like in shape with a mean particle size of 54.12 nm,polydispersity index of 0.122,Zeta potential of -13.60 mV;the linear range of puerarin was 2-10μg/ml(R2=0.999 4)with average recovery rate of 99.2%(RSD=0.9%,n=3). The re-covery rate of free drug was 95.3%(RSD=1.7%,n=3). The mean entrapment efficiency and drug-loading amount of puerarin were(35.5±2.12)% and(0.3±0.07)%,respectively(n=3). CONCLUSIONS:Film dispersion method is suitable for the prepara-tion of Puerarin polymeric micelles. Established method is convenient,accurate and reliable for the content and entrapment efficien-cy determination of Puerarin polymeric micelles.
10.Clinical study of pancreaticoenterostomy by separating and continuous suture in pancreatoduodenectomy
Gangcheng WANG ; Guangsen HAN ; Yingjun LIU ; Yong CHENG ; Yingkun REN ; Xianzhi LU ; Ping HUANG
Chinese Journal of Pancreatology 2016;16(1):19-22
Objective To investigate the effects of pancreaticoenterostomy by separating and continuous suture on pancreatoduodenectomy.Methods The clinical data of 76 patients who underwent curative pancreaticoduodenectomy from March 2002 to October 2014 in Tumor Hospital of Zhengzhou University were retrospectively analyzed.Of all the patients,43 received pancreaticoenterostomy by separating and continuous suture (study group),and the other 33 patients received BPJ anastomosis (control group).All the patients used Child reconstruction,the diagnosis of pancreatic fistula was made according to ISGPF criteria.The operation time of pancreaticoenterostomy,the incidence of hemorrhage of anastomosis and the incidence of pancreatic fistula were retrospectively analyzed.Results The age,sex,hemoglobin,albumin,total bilirubin,the incidence of co-morbidity of diabetes,extent of surgical resection between 2 groups were comparable without significant difference.In study group,the time of pancreaticoenterostomy was 11 min(8 ~ 15 min),there were 4 patients with class Ⅰ pancreatic fistula,and 1 patient with class Ⅱ pancreatic fistula.No patient developed class Ⅲ pancreatic fistula or anastomotic bleeding.In control group,the time of pancreaticoenterostomy was 16 min(12 ~25 min) which was only available for 5 patients,and no records for other patients.There were 6 patients with pancreatic fistula including 4 patients with class Ⅱ,2 patients with class Ⅲ,while no records for class Ⅰ.Four patients were found to have anastomotic bleeding.Conclusions With the pancreaticoenterostomy by separating and continuous suture method,the surgical field is fully exposed,the suture time is shortened and the incidence of anastomotic bleeding and pancreatic fistula is reduced.