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.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.
3.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.
4.Relationship between thyroid hormones and renal function after kidney transplantation
Juan LIU ; Wei LIU ; Daihong LI ; Lili XIE ; Qiang GAO
Chinese Journal of Organ Transplantation 2013;(1):9-12
Objective Monitoring the thyroid hormone levels in patients before and after renal transplantation to realize the relationship between thyroid hormones and renal function.Methods Fifty-seven patients were subdivided into stabled graft function group (50 cases) and delayed graft function (DGF) group (7 cases).Thirty healthy men served as control group.Serum triiodothyronine (T3),thyroxine (T4),thyroid-stimulating hormone (TSH) and serum creatinine (Scr) were determined respectably before transplantation and on different days after transplantation.Correlations between thyroid hormones and Scr at 10th day after transplantation were analyzed.Results Serum T3 and T4 levels before transplantation in the two groups were significantly lower than those in the control group (P<0.01).These four parameters levels had no significant difference between stabled graft function group and DGF group (P>0.05).In stabled graft function group:T3 level was decreased obviously by 30% in the 1st day after transplantation,which was higher than the others,elevated at the 1st week post-transplantation,reached the normal range.at the 2nd week post-operation,and higher than pre-transplantation (P < 0.01) at 3rd week; T4 revealed a decrease trend after transplantation and was elevated at 10th day post-transplantation,returned to the pre-transplantation level at 3rd week; TSH had slight decrease after operation,and had no significant difference among every period in comparing with pre-transplantation; Scr revealed a decrease trend continuously and returned to the normal level at the 7th day post-transplantation.In DGF group,Hormones levels were obviously decreased as compared with stabled graft function group; T3 reached the normal range at the 3rd week post-transplantation,whereas T4 returned to pre-transplantation levels at the 30th day; TSH had a significant decrease at the 5th day,and began to increase at the 10th day post-operation; Scr retuned to the normal range at the 30th day post-operation.The correlation coefficients between Scr and T3,T4 were 0.546 and 0.423 respectively.Conclusion There is a significant correlation between thyroid hormones and renal function (T3,T4).Monitoring the thyroid hormones,specially T3,can diagnose renal function change.
5.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.
6.Assessment of renal function in patients with renal transplantation by monitoring serum cystatin C
Daihong LI ; Wenli SONG ; Qiang GAO ; Juan LIU
Chinese Journal of Organ Transplantation 2010;31(7):425-428
Objective To evaluate the value of monitoring serum cystatin C in assessment of renal function in patients with renal transplantation.Methods Serum cystatin C, creatinine (SCr), β2-microproglobin (β2-MG) and urea nitrogen (BUN) levels were determined at different time points (pre- or post-operation) in 58 renal transplant patients.Glomerular filtrated rate (GFR) was determined by using of 99mTc-DTPA at the seventh day after operation.The correlation between GFR and the four markers was analyzed.Diagnostic characteristics and ROC curve for the four markers were obtained using a GFR cut-off of 1.5 ml/s.Intra-individual coefficients of variation (CV) for cystatin C and SCr according to different time points during post-operation monitoring and the ratio (R) between CVSCr and CVcystatin C were calculated.Results Cystatin C was decreased by 48.1 % at the first day after operation, which was higher than others.The correlation coefficients between GFR and cystatin C, SCr, β2-MG, and BUN were 0.876, 0.691, 0.589, 0.516 respectively.Diagnostic characteristics for GFR and cystatin C, SCr, β2-MG, and BUN were as follows:sensitivity (91.3 %, 87.2 %, 82.6 %, 87.0 %); specificity (80.0 %, 69.2 %, 71.4 %, 42.9 %), positive predictive value (82.0 %, 73.7 %, 74.3 %, 60.4 %); positive likelihood rate (4.81, 2.83, 2.87, 1.53).The area under the curve (AUC) for GFR and cystatin C, SCr, β2-MG, and BUN was 0.914, 0.828, 0.803, and 0.765 respectively.The CVSCr was significantly lower than CVcystatin C (P< 0.01).R was less than 1 in most patients with cystatin C<2 mg/L.In patients with cystatin C>2 mg/L, R tended value 1 with increasing concentrations of cystatin C.Conclusion Cystatin C showes the best correlation to GFR, and is superior to the other markers in accurate in differentiate mild renal impairment from moderate and severe renal impairment.When renal function has minimal change, the cystatin C level has significant change.When the renal function has mild impairment, great changes in serum cystatin C indicate the unstable renal function.
7.Analysis on predictive factors of survival to in-hospital children with post-cardiorespiratory arrest
Yi XIN ; Weina SONG ; Qing CHU ; Daihong WAN ; Aimin LI
Chinese Pediatric Emergency Medicine 2010;17(1):26-29
Objective To identify predictive factors of survival to in-hospital children with post-cardiorespiratory arrest.Methods Eighty-seven patients who had cardiorespiratory arrest(CRA)and received cardiopulmonary resuscitation(CPR)were reviewed from January 2006 to December 2008.The clinical data was analyzed to identify predictive factors of survival by Pearson X~2 test and multivariate and unconditioned Logistic regression analysis.Results Forty-three of eighty-seven cases received return of spontaneous circulation,the initial sunvival rate was 48.3%,31(35.6%)cases survived for 24 hours,19(21.8%)cases survived to hospital discharge.Primary diseases and complications,the type of arrest,tracheal intubation or not,the duration of CPR,the dose of adrenaline,the body temperature post-resuscitation for 24 hours,the glucose post resuscitation for 6 hours and complications were obviously associated with both initial and long-term survival.Logistic regression analysis revealed that primary diseases and the duration of CPR were predictive factors of both survival for 24 hours and hospital discharge,additionally,the body temperature post-resuscitation for 24 hours were predictive factor of survival to hospital discharge.Conclusion Initial and long-term survival rate of in-hospital children with post-resuscitation was both low.Primary diseases and complications,good quality CPR and management of post-resuscitation affect survival,especially primary diseases and the duration of CPR may predict initial survival and primary diseases,the duration of CPR and the body temperature post-resuscitation for 24 hours may predict long-term survival.
8.Study on the role of serum cytokine in the patients of HBV recurrence after liver transplantation.
Hongli SONG ; Zhongyang SHEN ; Daihong LI ; Al ET ;
Chinese Journal of Practical Internal Medicine 2006;0(S2):-
Objective To evaluate the role of cytokine levels for prediction of HBV recurrence after liver transplanta- tion.Methods 34 cases of liver transplantation from June 2002 to December 2003 in our hospital were observed.The measurements of cytokine in serum by ELISA method were taken on the 7 days before operation and recurrence after oper- ation and the change rules were studied.Results The serum TNF-?,IL-10和 INF-? levels gradually increased in HBV non-recurrence group,but which in recurrence group had no markedly change and always skept on normal levels after operation.In addition,the serum TNF-?和 IL-10 levels after operation were significantly higher in nonrecur- rence group than in recurrence group.The serum IL-2 levels in recurrence group always kept on higher levels without marked changes after operation.Conclusion TNF-? and IL-10 are better laboratory index in the prediction of HBV recurrence after liver transplantation and which are noninvaded and cheap.
9.Effect of intravenous immunoglobulin on immunity and outcome for sepsis in children
Yi XIN ; Daihong WAN ; Qing CHU ; Qi ZHAO ; Xingjuan GAO ; Aimin LI
Chinese Pediatric Emergency Medicine 2011;18(6):497-499
Objective To explore the effect of intravenous immunoglobulin (IVIG) on immunity and outcome for sepsis in children.Methods Eighty-four children who met the diagnosis of sepsis were included in study and divided into treatment group (36 cases) and control group (48 cases ).The patients in teatment group were administered IVIG with the dose of 1 g/kg.Peripheral venous blood samples of patients in both groups were collected before (0 h),24 h,72 h and 5 d after administration to detect the numbers of immunocyte including CD3 +,CD4 +,CD56 +,CD19 +,CD8 +cells by flow cytometry and the levels of cytokines including tumor necrosis factor (TNF)-at,interleukin (IL)-10,IL-1 7 by enzyme linked immunosorbent assay.The numbers of immunocyte and levels of cytokines and TNF-a/IL-10 were compared and the mortality at 28 days was assessed between two groups.Results The numbers of CD3 +,CD4 +,CD56 +,CD19 +cells and the levels of TNF-a,IL-17 and TNF-α/IL-10 of patients in teatment group were significantly decreased than those in control group at 24 h,72 h and 5 d afte administration ( P <0.05 ) and showed downtrend.However,the level of IL-10 increased significantly (P < 0.05 ) and showed uptrend in treatment group.The number of CD8+ cells had no change.No difference of mortality was observed between two groups (27.7%,10/36 vs 16.6%,8/48,x2 =1.50,P =0.169,OR =1.92,95% CI:0.671 ~5.510).Conclusion IVIG can suppress the immunity of children with sepsis and has no survival benefit.
10.Clinical application of Cylex ImmuKnow assay in patients with renal dysfunction after liver transplantation
Weiping ZHENG ; Daihong LI ; Yihe LIU ; Hong ZHENG ; Yonglin DENG ; Cheng PAN ; Zhongyang SHEN
Chinese Journal of Organ Transplantation 2013;34(11):661-665
Objective To assess the clinical application of Cylex ImmuKnow assay in patients with renal dysfunction after liver transplantation for individualized immunosuppressive therapy.Method Twenty adult patients undergoing liver transplant between January 2009 and December 2011 received regular ImmuKnow assay monitoring combined with determination of serum tacrolimus trough concentration to guide immunosuppressive regimens,all of whom showed sustained renal dysfunction 6 months after transplant with normal and stable liver function.Clinical data were collected to observe the changes of renal function in those patients after treatment.Results The recipients were followedup for 15-54 months,received ImmuKnow assay 61 times and the results fluctuated 33-943 μg/L [median 282 μg/L,interquartile range (IQR) 267 μg/L].After ImmuKnow monitoring,serum creatinine level in patients was decreased significantly from median 151.8 μmol/L with IQR 44.9 μmol/L to median 114.9 μmol/L with IQR 35.3 μmol/L (Z =-3.845,P =0.000),and estimated glomerular filtration rate (eGFR) was increased significantly from median 0.746 mL/s with IQR 0.025 mL/s to median 1.005 mL/s with IQR 0.454 mL/s (Z =-3.771,P =0.000).ImmuKnow results showed a linear correlation with the white blood cell count in patients (Spearman correlation coefficient r =0.429,P =0.001),but no linear correlation with the patients' age,primary disease before transplantation,postoperative time,serum tacrolimus trough concentration,lymphocyte count,CD3+ T lymphocyte count,CD4+ T lymphocyte count or CD4+/CD8+ T lymphocyte ratio (P> 0.05).Conclusion Cylex IrmmuKnow assay can be applied in patients with renal dysfunction after liver transplantation for individualized immunosuppressive therapy monitoring,which is of certain clinical value.