1.The role of bone morphogenetic protein-7 (BMP-7) in inducing expression of extracellular matrix and nuclear factor-?? in proximal tublar epithelial cells by high glucose level
Daihong WANG ; Li LIU ; Fahuan YUAN
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
Objective To investigate the effect of bone morphogenetic protein-7 (BMP-7) on high glucose-induced expression of fibronectin (FN) and collagen Ⅳ (Col Ⅳ) and activity of nuclear factor-?? (NF-??) in human renal tubular epithelial cells. Methods Human renal tubular epithelial cells (HKCs) in culture were divided into 5 groups: normal glucose group with 5.5 mmol/L glucose (NG), high glucose group with 25 mmol/L D-glucose (HG), HG+100 ng/ml BMP-7, NG+100 ng/ml BMP-7 and high osmolality group with 25 mmol/L mannitol (HM). Thus HKCs were respectively stimulated with high glucose, high mannitol and BMP-7. Expression of collagen Ⅳ and fibronectin was determined by immunocytochemistry and enzyme linked immunosorbent assay (ELISA) and activity of nuclear factor-?? was assessed with electrophoretic mobility shift assay (EMSA). Results Compared with normal glucose, high glucose concentration up-regulated expression of FN and Col Ⅳ and activity of NF-?? (P0.05). With addition of BMP-7 in a concentration of 100ng/ml to the high glucose medium, expression of Col Ⅳ and FN was suppressed, and the activity of nuclear factor-?? was inhibited in human tubular epithelial cells (P
2.Study on the value of the diagnosis and efficacy evaluation of c-reactive protein (CRP) test in patients clinical complications after bone marrow transplantation
Jing WANG ; Lanping XU ; Daihong LIU
Chinese Journal of Practical Internal Medicine 2001;0(06):-
Objective To study the value of c-reactive protein (CRP) level in the diagnosis and efficacy evaluation of clinical complications in patients after bone marrow transplantation.Methods Sixty-six patients with different complications after bone marrow transplantation were included and CRP levels were tested.The relationship between CRP levels and corresponding clinical complications were analyzed.Results The levels of CRP were significantly different in different complications after the bone marrow transplantation.CRP may marketly increase in the patients with bacterial infection,bacteria-associated mixed infection,ultra-acute GVHD,and ATG reaction;CRP may mildly increase in patients with bacterial infection,fungal infection,bacteria-associated mixed infection,acute/chronic GVHD;CRP did not increase in the patients with viral infection.CRP level reached the highest value in patients with most obvious clinical symptoms of bacterial infection,gradually decreased in patients after effective treatment,those restored to normal but then increased CRP always indicated new clinical complications.Conclusion CRP is a valuable mark for the early differential diagnosis and efficacy of therapy for complications in patients after bone marrow transplantation.
3.The inhibitive effect of PTEN protein on secretion of collagen Ⅳ and fibronectin by stimulation of by TGF-?_1 in rat
Yue CHENG ; Daihong WANG ; Yaoquan ZHANG
Medical Journal of Chinese People's Liberation Army 2001;0(07):-
Objective To investigate the role of PTEN protein in secretion of collagen Ⅳ and fibronectin after stimulation of transforming growth factor beta 1(TGF-?1) from renal fibroblasts of rat in vitro.Methods The cultured rat renal fibroblasts were transfected with the reconstructed adenovirus containing PTEN or adenovirus only containing green fluorescence protein(GFP).The fibroblsts were treated in four manners:control group with no added treatment,TGF-?1 group with TGF-?1 stimulation,PTEN+ TGF-?1 group with TGF-?1 stimulation after Ad-PTEN transfection,and GFP+ TGF-?1 group with TGF-?1 stimulation after Ad-GFP transfection.Invert fluorescent microscope was used to detect the GFP expression,meanwhile the PTEN mRNA was determined by RT-PCR method.36h after the transfection,TGF-?1 was added into the culture medium in a concentration of 10ng/ml.After another 24h,ELISA method was used to evaluate the level of collagen Ⅳ and fibronectin.Results The expressions of both GFP and PTEN mRNA increased obviously after the rats' renal fibroblasts were transfected with adenovirus.The secretion of collagen Ⅳ and fibronectin increased significantly in both TGF-?1 group and GFP+ TGF-?1 group compared with that in control group,and decreased markedly in PTEN+ TGF-?1 group compared with that in both TGF-?1 group and GFP+ TGF-?1 group(P
4.The clinical study of non-T-depleted human leukocyte antigen mismatched hematopoietic stem cell transplants with total body irradiation for the treatment of leukemia
Yu WANG ; Lanping XU ; Daihong LIU
Chinese Journal of Practical Internal Medicine 2006;0(20):-
20.0?109/L was day 13(11~21).All the patients got engraftment successfully and attained CR.Acute Ⅰ~Ⅱ GVHD occurred in 5 patients,no acute Ⅲ~Ⅳ GVHD occurred and extensive chronic GVHD did in 2 patients.All the patients were alive and only one patient relapsed after 3~53 months follow-up.Conclusion TBI-based conditioning regimen appears to be safe and effective for non-T-depleted HLA mismatched hemotopoietic stem cell transplants,especially for high-risk and second transplant patients.
5.Effect of mast cell tryptase on PAR-2 and MCP-1 mRNA expression of human renal fibroblasts in culture
Daihong WANG ; Fahuan YUAN ; Yaoquan ZHANG
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective To further investigate the effect of mast cell tryptase on the pathogenesis of chronic renal fibrosis. Methods Renal interstitial fibroblasts were isolated from histologically normal renal tissue obtained from kidneys removed because of malignant renal tumor by collagenase disintegration, and cultured in vitro. The cultured fibroblasts were identified by cell shape and immunocytochemistry, and divided into groups for further experiments. The expression of monocyte chemoattractant protein-1 (MCP-1) and protease-activated receptor 2(PAR-2) mRNAs in in vitro cultured fibroblasts was assessed by means of semi-quantitive reverse transcriptase polymerase chain reaction (RT- PCR). Result Renal fibroblasts were successfully cultured. Mast cell tryptase(10~500ng/ml) promoted expression of PAR-2 mRNA and MCP-1 mRNA of human renal fibroblasts in heparin-dependent manner. Proteinase inhibitor benzamidine hydrochloride hydrate (200?mol/L) could suppress the effects of tryptase on the cultured renal fibroblasts. But TGF-? antibody did not influence the effect. Conclusion Tryptase might be involved in pathogenesis and development of renal fibrosis by PAR-2 and MCP-1.
6.Practice of continuous quality improvement in infection control of blood purification center
Yiqin WANG ; Daihong WANG ; Jianying TANG ; Hui LI ; Fahuan YUAN
Chinese Journal of Medical Education Research 2011;10(3):358-359
Infection control is important in quality management of blood purification center. Practice of Continuous Quality Improvement in infection control improves patients' living quality, and it is worth proceeding and generalization.
7.Protective effect of shenfu injection on intestinal mucosal ischemia-reperfusion injury and intestinal epithelial cells at recovery phase in rats
Daihong WANG ; Xiaofei HE ; Xiangchu LI ; Chaoxin ZHENG ; Xunchen ZHANG
Chinese Journal of Tissue Engineering Research 2006;10(47):202-205
BACKGROUND: Apoptosis plays a key role in intestinal mucosal ischemia-reperfusion injury and recovery; meanwhile, effect of shenfu injection on apoptosis of intestinal epithelial cells during intestinal mucosal ischemia-reperfusion injury should be studied further.OBJECTIVE: To investigate the relationship between the apoptosis of intestinal epithelium and characteristics of intestinal mucosal ischemia-reperfusion injury and recovery.DESIGN: Randomized controlled animal experiment.SETTING: Department of General Surgery, Xianning Central Hospital;Department of General Surgery, Renmin Hospital of Wuhan University.MATERIALS: The experiment was carried out in the Central Laboratory,Xianning Central Hospital from March to August 2005. Fifty-four healthy male SD rats weighting 200-250 g were provided by Animal Center of Medical School, Wuhan University.METHODS: The rats were divided randomly into 3 experimental groups:control group (n=6), ischemia-reperfusion group (n=24) and shenfu treatment group (n=24). ① Pentobarbital sodium solution (40 mg/kg) was administrated into the intraperitoneal cavity to induce anaesthesia. Through a midline abdominal incision, the mesenteric blood vessel of a 15-cm segment of mid-intestine was occluded for 60-minute with an atraumatic vascular forceps. The control group underwent the same procedure except for unblocking the mesenteric blood vessel. At the end of 60 minutes ischemia period the forceps was removed to allow reperfusion, the abdominal cavity was closed. ShenFu injection (8 mL/kg ·h, 20 mL/kg ·d, produced from Yaan Three-Nine Pharmaceuticals Co, No: 030302) was injected 30 minutes before occlusion in SF treatment group, same quantity of 0.9% natrii chloride was injected in control group and ischemia-regeneration group at the same time, and oxygen was inbreathed during the operation and ischemia-regeneration. ② Experimental intestinal canals were sampled for the following analysis when all groups were respectively performed sham ischemia for 1 hour, intestinal ischemia for 1 hour and reperfusion for 1, 24and 72 hours. Sections were observed in light microscope. Histological mucosal damage in each sample was evaluated as followed scoring system: 0score, normal muscosal villi and gland; 1 score, slight lesion near the tip of the villi; 2 scores, slight lesion of subepithelial gland; 3 scores, development of subepithelial (Gruenhagen) spaces near the tip of the villi with capillary congestion; 4 scores, extension of the subepithelial space with moderate epithelial lifting from the lamina propria; 5 scores, a few denuded villous tips; 6 scores, massive denuded villi; 7 scores, denuded villi with exposed lamina propria and obvious gland lesion; 8 scores, disintegrateon of the lamina propria; 9 scores, haemorrhage and ulceration. ③ The Tunel method (TdT mediated biotin-dUTP nick and labeling; TdT-Frag EL DNA fragmentation detection kit) was used. Inbrief, this method allowed the identification of apoptosis nuclei in tissue samples through DNA fragment and labeling. Apoptosis Index (AI) was set as the average number of apoptosis cells in per 100 cells by observing ten high power fields of adjacent villi and crypts. ④ The mitotic phase of crypt epithelial nucleus within intestinal mucosa was observed in intestinal sections stained with haematoxylin and eosin. The number of cells with nucleus mitotic phase was counted in ten adjacent mucosal crypts, which was taken as the index of mitotic activity of intestinal mucosal epithelial cell.MAIN OUTCOME MEASURES: Intestinal mucosal histopathological changes, apoptosis of intestinal mucosal epithelial cell and mitotic activity of intestinal mucosal crypt.RESULTS: All 54 rats were involved in the final analysis. ①) Scores of histopathological changes were (0.65 ±0.35) points in 1-hour ischemia group, (3.87±0.86) points in 1-hour reperfusion group and (0.65±0.35)points in 24-hour reperfusion group; which were lower than those in ischemia-reperfusion group [(7.11±1.01), (8.05±1.34), (1.53±0.48) points; P< 0.05]. ② Indexes of apoptosis were 17.24±7.05 in 1-hour ischemia group, 24.20±9.87 in 1-hour reperfusion group, 11.49±4.71 in 24-hour reperfusion group and 6.02±2.16 in 72-hour reperfusion; which were lower than those in ischemia-reperfusion group (51.09±13.76, 54.89±15.58,23.54±9.64, 12.47±5.52; P < 0.05). Activities of mitosis were 10.37±2.03and 11.72±2.07 in 1-hour ischemia group and 1-hour reperfusion group,respectively; which was higher than those in ischemia-reperfusion group(8.24±1.69, 9.95±1.93; P < 0.05).CONCLUSION: Shenfu injection can significantly attenuate apoptosis of intestinal epithelium, increase crypt mitotic activity, and promote intestinal epithelium regeneration or repair.
8.Correlation Analysis between Ciclosporin A and Biochemical Indicator of Blood in Renal Transplantation Patients
Hong WANG ; Daihong GUO ; Yihong CHEN ; Baoxiang PEI
China Pharmacy 1991;0(05):-
OBJECTIVE:To analyse the correlation between cyclosporine A(CsA)and the biochemical indicator of renal transplantation patients.METHODS:The patients were divided into6groups according to the different time length after renal transplatataion:0y~1y,1y~2y,2y~3y,3y~4y,4y~5y and above5y;the blood samples were taken12h after oral administra?tion of CsA,the of concentration CsA in blood of each group was determined and the biochemical tests of which were taken;post-operation medication and the correlation between the blood biochemical index were statistically analyzed by Stata soft?ware.RESULTS:The dosage of CsA decreased gradually from group1to group6,and the concentration of CsA also de?creased.And the data of BUN in group1,5and6were higher than the upper limit of their standard data,the data of triglyc?eride(TG)in group1,2,3,5and6were higher than the upper limit of their stand data;there were significant differences in the data of TP、ALB、TB、DB、Ua、TC、TG、?-GT among the6groups,while the data of ALP、ALT、AST、Glu、BUN、Cr among the6groups have no notable difference.CONCLUSION:The administration of CsA after renal transplantation can change bio?chemical indicator in patients.
9.Effects of D-cresol on the proliferation and cell cycle of human umbilical vein endothelial cells
Li LI ; Lixia GUANG ; Daihong WANG ; Xiang DU ; Fahuan YUAN
Chinese Journal of Nephrology 2013;29(9):676-680
Objective To investigate the effects of p-cresol on human umbilical vein endothelial cells.Methods The effects of p-cresol on endothelial cell growth,cell cycle,cell morphological change and p21 protein were detected by the CCK-8 assay,flow cytometry assay,inverted microscope and Western blotting.Results P-cresol could inhibit the growth of human umbilical vein endothelial cells in dose-and time-dependent manners (all P < 0.05).The human umbilical vein endothelial cells treated with p-cresol became elongated processes,cloudy cytoplasm,and irregular shapes.The p-cresol stopped human umbilical vein endothelial cells at cell cycle G1 and had no effect on cell apoptosis.The p-cresol could increase protein expression of p21 in a dose dependent manner (P < 0.05).Conclusion P-cresol can increase protein expression of p21,induce cell cycle arrest at G1 stage and inhibit the proliferation of human umbilical vein endothelial cells.
10.Antidepressant effects of piperine and its neuroprotective mechanism in rats.
Yuan HU ; Hongbo LIAO ; Ping LIU ; Daihong GUO ; Yuyu WANG
Journal of Integrative Medicine 2009;7(7):667-70
To study the antidepressant effects of piperine in chronic unpredictable mild stress (CUMS) rats and to explore the underlying mechanisms in the hypothalamic-pituitary-adrenal (HPA) axis.