1.Effect of CXCR4-overexpressing bone marrow-derived mesenchymal stem cells on the repair of the co-cultured hypoxia/re-oxygenation renal tubular epithelial cells and its possible mechanism
Nanmei LIU ; Changlin MEI ; Jinyuan ZHANG ; Jun TIAN ; Jin CHENG
Chinese Journal of Nephrology 2013;29(11):830-836
Objective CXCR4-overexpressing bone marrow-derived mesenchymal stem cells (CXCR4-BMSC) were constructed and co-cultured with hypoxia/re-oxygenation pretreated renal tubular epithelial cells (HR-RTEC).Repair of HR-RTEC was detected and the possible mechanism was also discussed.Methods CXCR4-BMSC (CXCR4-BMSC/eGFP,eGFP as the tracer gene) and null-BMSC (BMSC/eGFP) were obtained by gene transfection technique,and the level of CXCR4 in the transfected cells was detected.RTEC was cultured under hypoxia/re-oxygenation condition for 12 h,respectively,to obtain HR-RTEC,which was used to simulate AKI in vitro.BMSC and HR-RTEC were co-cultured for 12 h,and the proportion of apoptotic cells among the HR-RTEC was assayed by immunofluorescence technique.Western blot was used to test the protein levels of cleaved Caspase-3 and Bcl-2.The number of migrating BMSC was also assayed.After culturing with the HR-RTEC culture supernatant,the expression of cytokeratin 18 (CK18) in BMSC was tested by immunofluorescence staining.Cytokines including bone morphogenetic protein-7 (BMP-7),hepatic growth factor (HGF) and interleukin-10 (IL-10) in the BMSC culture supernatant were detected by ELISA method.Results Expression of CXCR4 was enhanced in CXCR4-BMSC.Proportions of the apoptotic cells among HR-RTEC after being co-cultured with BMSC,CXCR4-BMSC and null-BMSC were all decreased,especially in the C/H group.The decreased cleaved Caspase-3 and enhanced Bcl-2 were also observed in HR-RTEC.The number of migrating CXCR4-BMSC was the highest.Proportions of CK18+ cells in BMSC,CXCR4-BMSC and null-BMSC were all low and showed no difference.However,CXCR4 overexpression in BMSC stimulated secretions of BMP-7,HGF and IL-10.Conclusions CXCR4-overexpressing BMSC has more repair effect on the co-cultured HR-RTEC,the enhanced migration ability and secretion ability of CXCR4-BMSC are the possible mechanisms.
2.Celastrol down-regulates expressions of MIF and MMP-9 in the aorta of ApoE gene knockout mice
Jinping LI ; Jun CHENG ; Zhuo TIAN ; Houyuan HU
Journal of Third Military Medical University 2003;0(09):-
Objective To investigate the effects of celastrol on the expressions of macrophage migration inhibitory factor(MIF)and matrix metalloproteinase-9(MMP-9)in the aorta of apoE gene knockout mice with earlier atherosclerosis.Methods Eight-week-old ApoE gene knockout male mice were divided randomly into control group and celastrol treatment group(n=6 in each group).The mice in celastrol group were given.celastrol(2 mg?kg-1?d-1)by intraperitoneal injection for 4 weeks;and the mice in control group were only given equivalent amount of dimethyl sulfoxide(DMSO).HE staining of root aorta were used to observe the histomorphological changes and measure the size of plaque in ApoE-/-mice.The expressions of MIF and MMP-9 were detected by immunological histochemical method.Results The area of lipid plaque in the mice treated with celastrol was significantly smaller than that of the control(P
3.Effects of liver X receptor agonist on the expressions of C-reactive protein and CD40 ligand and contents of smooth muscle cells in the aorta of ApoE gene knockout mice
Jun CHENG ; Jinping LI ; Zhuo TIAN ; Houyuan HU
Journal of Third Military Medical University 2003;0(13):-
Objective To investigate the effects of liver X receptor agonist on the expressions of C-reactive protein and CD40 ligand and smooth muscle cell ?-actin in the aorta of ApoE gene knockout mice with earlier atherosclerosis. Methods Male ApoE gene knockout mice (8-week old) were divided randomly into control group and T0901317 treatment group (n=6 in each group). The mice in T0901317 group were administered intraperitoneally with T0901317 at the dose of 20 mg?kg-1?d-1 for 4 weeks. Mice in the control group were only given equivalent amount of dimethyl sulfoxide (DMSO). The expressions of C-reactive protein and CD40 ligand and smooth muscle cell ?-actin were detected by immunological histochemical method. Results The expressions of C-reactive protein and CD40 ligand in the atherosclerotic plaque in the aortic wall were significantly lower in T0901317 group as compared with those in the DMSO control group (P0.05). Conclusion Liver X receptor agonist may reduce the formation of atherosclerotic lesions by inhibiting the inflammation and the expressions of C-reactive protein and CD40 ligand in the aorta of ApoE gene knockout mice.
4.Study on Polyploid Induction and Identification of Nervilia fordii
Yufeng LIN ; Qin DU ; Fengli CHENG ; Zhenhua WANG ; Jun TIAN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(3):513-518,579
Objective To investigate the polyploid induction and identification of Nerviliae fordii for harvesting the polyploid plants. Methods The materials and methods for polyploid induction of Nerviliae fordii were screened separately by comparing the induction rates of rhizomes and bulbs under natural conditions and tissue culture environment, and by comparing soaking method with agar method. The effects of colchicine concentration ( 200, 300, 400, 300 mg/L), colchicine action time ( 7, 14, 21, 28 d), DMSO concentration ( 0, 10, 20, 40 mL/L) and KT concentration ( 0, 1.0, 2.0, 4.0 mg/L) on induction rate were observed by orthogonal design method. The polyploid induction in the treated plants was identified by morphology, cytology and chromosome methods. Results After the tissue culture rhizomes were treated with 300 mg/L colchicine, 10 mL/L DMSO, and 2.0 mg/L KT by agar method for 28 days, the polyploid induction rate arrived to 50%, showing better induction effect. The morphology of polyploid plants was characterized by giantism, and the leaf length, leaf width and plant height were respectively 152.17%, 158.67%and 60.90%of those of the diploid plants. The length, width and density of stoma of leaf epidermal cell as well as the number of chloroplast in the treated plants were 138.46%, 153.00%, 59.09% and 109.09% of those of the untreated plants. The results of chromosome identification showed that the amount of the tetraploid ( about 40) was 2 times of the diploid chromosome ( about 20) in the treated plants, proving that the achieved Nerviliae fordii was a tetraploid plant. Conclusion Polyploid plants of Nerviliae Fordii have been successfully obtained, which will supply evidence for improving species, richening seed-breeding resources, and selecting of improved seeds of Nerviliae fordii.
5.CELL IMMOBILIZATION FOR THE USE OF WASTEWATER TREATMENT
Yali CAO ; Shen TIAN ; Jun ZHAO ; Cheng QIAN ; Xiu YANG ;
Microbiology 1992;0(03):-
Cell immobilization is a new biotechnology The definition, classification, and carrier selection of cell immobilization are presented in details The technique is efficiently applied to treating strength organic wastewater, nutrient and heavy metals removal of wastewater, as well as hardly biodegradated wastewater It has a widely applied prospect in wastewater treatment
6.Effect of erythropoietin on the oriented chemotaxis of bone-marrow mesenchymal stem cells under acute kidney injury microenvironment
Nanmei LIU ; Changlin MEI ; Jinyuan ZHANG ; Jun TIAN ; Jin CHENG ; Weiwei WANG
Chinese Journal of Nephrology 2013;(4):263-267
Objective To investigate the migration of bone-marrow mesenchymal stem cells (BMSCs) under acute kidney injury (AKI) microenvironment in vitro and the effect of erythropoietin (EPO) intervention,and to explore its underlying mechanism.Methods Renal tubular epithelial cells (RTECs) were cultured in hypoxia/re-oxygenation (HR) condition for 12 h,respectively,in order to establish HR-RTEC.BMSCs and RTECs were co-cultured by Transwell system and were divided into 7 groups:control group (group①,only BMSC cultured),BMSC-RTEC co-culturing group (group ②),BMSC-HR-RTEC co-culturing + EPO intervention groups (group ③to group ⑦,EPO concentration:0,1,5,10,50 IU/ml).All the groups were cultured for 48 h and the number of migrating BMSCs was detected.Western blotting was applied for the detection of SDF-1 expression in RTECs and pMAPK and MAPK levels in BMSCs.SDF-1 concentration in the RTECs culture supernatant was tested by ELISA.Results The number of BMSCs migrating to the low chamber where HR-RTECs were cultured was increased,and EPO intervention further enhanced this migration which reached the peak at the concentration of 10 IU/ml [Compared with group③,(46.67±7.37) cells vs (19.00±2.37) cells,P < 0.05].Intracellular expression level and the secreated level of SDF-1 in HR-RTECs in group③ were higher than those in RTECs of group② [0.37±0.01 vs 0.19±0.01,P < 0.05; (61.64±4.88) μg/L vs (35.26±8.78) μg/L,P < 0.05].EPO intervention increased above SDF-1 levels and reached the peak at the concentration of 10 IU/ml [group⑥ vs group③:(173.53± 14.66) μg/L vs (61.64±4.88) μg/L,P<0.05],accompanied with enhanced phosphorylation of MAPK in BMSCs.Conclusions AKI microenvironment has obvious chemotaxis effect on BMSCs,and EPO intervention can strengthen this effect.The increased SDF-1 level and enhanced phosphorylation of MAPK,the downstream signal protein of SDF-1/CXCR4 axis,are the possible mechanism for EPO performance.
7.mRNA expression change of myelin gene in hippocampus following cerebral ischemia
Ying-Zhu CHEN ; Huan BAO ; Ye TIAN ; Shi-Yao BAO ; Jun XU ; Cheng-Lin YUAN ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To study the mRNA expression of myelin basic protein (MBP) and myelin oligodendregha glyeoprotein (MOG) in hippocampus of rats following global brain ischemia.Method The four- vessel occlusion animal model in the Sprague-Dawley rats was used in this study.The mRNA expression levels of MBP and MOG in the hippocampus of rats were analyzed by reverse transcription polymerase chain reaction (RT- PCR) at day 2,4,7,14 and 28 days after global brain ischemia.There were eight rats at each time-point and sham operated group.Results The mRNA expression of both MBP and MOG in hippocampus of rats decreased at 2 days after global brain ischemia.The gene expression of myelin gene decreased significantly at 7 days and it reached to the lowest level at 28 days.Compared with sham operated group,the gene expression of MBP and MOG in hippocampus of rats decreased significantly at 7,14 and 28 days after global brain ischemia (P
8.Effects of glucagon-like peptide 2 on recovery of small intestinal epithelia from radiation injury in mice
Jun-Dong ZHU ; Yong-Ping SU ; Tian-Min CHENG
Journal of Third Military Medical University 2001;23(3):293-295
Objective To investigate the effects of glucagon-like peptide 2 (GLP-2) on recovery of small intestinal epithelia from radiation injury in mice. Methods Mice received a single 8 Gy dose of total body irradiation from 60Co gamma ray followed by treatment with GLP-2 or vehicle. DNA and protein content in small intestinal mucosa were measured, and small intestine was processed for histological examination with light microscope and scanning electron microscope. Results Small intestinal mucosal DNA and protein content, villus height, and villus number significantly decreased in irradiated mice, partial villus tips were ulcerated. GLP-2 administration caused increase in DNA and protein content, villus height, and villus number as compared with irradiated control group. Meanwhile, the villus tips were lack of ulceration. Conclusion GLP-2 can promote recovery of small intestinal epithelia from radiation injury in mice.
9.Effect of glucagon-like peptide 2 on mitogen-activated protein kinase activity in small intestinal epithelia of mice after radiation injury
Jun-Dong ZHU ; Yong-Ping SU ; Tian-Min CHENG
Journal of Third Military Medical University 2001;23(4):375-377
Objective To study the effect of glucagon-like peptide 2 (GLP-2) on mitogen-activated protein kinase (MAPK) activity in small intestinal epithelia in mice after radiation injury and its relation with the change of small intestinal epithelial proliferation. Methods Mice were given a single dose of 8 Gy of total body 60Co gamma irradiation and then divided into GLP-2 and control groups. The activity of MAPK and proliferation rate in small intestinal epithelia were measured. Results The activity of MAPK in small intestinal epithelia was higher in GLP-2-treated mice than in irradiated mice, and the proliferation rate in small intestinal epithelia significantly increased in GLP-2-treated mice. These two indices were of significantly positive correlated. Conclusion GLP-2 can promote small intestinal epithelial proliferation in irradiated mice, and this may be related to activation of MAPK in small intestinal epithelia.
10.Resection for tumors of the suprarenal infrahepatic segment of the inferior vena cava
Jianfei CHEN ; Zhilei CHENG ; Yuanhu TIAN ; Jun CAI ; Chengli MIAO ; Chenghua LUO ; Jiye ZHU
Chinese Journal of General Surgery 2016;31(3):227-229
Objective To evaluate surgical treatment for different types of tumor of the suprarenal infrahepatic segment of the inferior vena cava (IVC).Methods A retrospective analysis was made on 8cases of IVC leiomyosarcoma at the suprarenal infrahepatic IVC segment.According to the extent of tumor invasion and collateral circulations,operations included tumor resection plus inferior vena cava reconstruction or tumor resection plus right nephrectomy.Results Tumors were completly resected in all patients.Renal function recovered after operation.During a follow up of 24 mons,7 cases had no recurrence.One case with recurrence after 12 months underwent successful resection and is doing well after a follow-up of 12 months.Conclusion CT and inferior vena cavography can determine the extent of tumor invasion and collateral circulations helping select appropriate surgical aproach before the operation resulting in less renal injury.