BACKGROUND:There are no unified methods to isolate,culture and amplify bone marrow mesenchymal stem cells(BMSCs) . Endothelial progenitor cells(EPCs) can be harvested by inducing peripheral blood or bone marrow mononuclear cells. However,the method of collecting bone marrow mononuclear cells is various,and the isolation efficiency is different. OBJECTIVE:To isolate and identify BMSCs and EPCs in vitro. DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Central Laboratory,First Affiliated Hospital,Anhui Medical University from October 2008 to February 2009. MATERIALS:Two clean healthy male Sprague Dawley rats were supplied by the Anhui Animal Center. METHODS:Rat bone marrow was extracted to harvest mononuclear cells by density gradient centrifugation. BMSCs were isolated by the differential adhesion. Following 48 hours of primary culture of BMSCs,non-adhered cells were collected and induced by EGM-2 complete medium supplemented with fetal bovine serum,vascular endothelial growth factor,fibroblast growth factor,insulin-like growth factor-1 and recombinant human epidermal growth factor. There were three groups:early adherence + common culture EPCs,twice adherence + induction culture EPCs,and rat mature aorta EPCs. Nitric oxide content was directly measured in the medium using nitrate reductase method. MAIN OUTCOME MEASURES:The following parameters were measured:in vitro culture of BMSCs and EPCs,and nitric oxide content in the medium. RESULTS:Primary culture of BMSCs was in a spindle-shape,within 24 hours the majority of cells were adherent,9-10 days up to 90% cells were confluent. Purified BMSCs were amplified and uniform,long spindle,with the passage cycle for 8 days. Flow cytometry detection showed that cells were CD34-,CD45-negative and CD105-positive. Second adherent of EPCs induced by 3 days culture showed colony formation following six days,appeared cord-like structure and microvessel-like growth at 8-10 days. At 2 weeks,the majority of cells were polygonal,colony-forming unit was interconnected,showing a typical "cobblestone"-like. At 7-10 days,cells were double stained DiI-acLDL and FITC-UEA-1. Flow cytometry showed Flk-1-positive and CD133-positive. Nitric oxide content was significantly higher in the culture medium compared with the ordinary cultured cells(P

AIM: The autologous bone marrow mesenchymal stem cell (BMSC) transplantation for treating acute myocardial infarction (AMI) via coronary artery in clinic has made remarkable and encouraging progress. The experiment aimed to study cytokine secretion in autologous BMSC transplantation for myocardial infarction dogs and its influence on angiogenesis and cardiomyocyte apoptosis. METHODS: Experiments were performed at the Animal Laboratory and the Central Laboratory, First Affiliated Hospital of Anhui Medical University between December 2005 and May 2007. 10 mL of bone marrow was sterilely aspirated from the posterosuperior iliac spine of 36 healthy mongrel dogs. BMSCs were isolated and purified by Percoll density gradient centrifugation and cultured in vitro by adherence method. After being co-cultured with 5-azacytidine for two passages, these cells were labeled by 5-bromodeoxyuridine (Brdu) for preparation. Dog models of AMI were established in a model control group and a cell transplantation group randomly. 2 mL of cell suspension of autologous BMSCs were implanted into four different regions in the acute myocardial site via topical injection in the cell transplantation group, while the same dose of DMEM was injected into the corresponding regions in the model control group. Myocardial infraction tissues were measured after transplantation. RESULTS: Flow cytometry demonstrated that the positive rate of CD34 and CD11b on third passage of BMSC cell surface was below 5%, while the positive rate of CD44 and CD105 was above 90%. In situ end labeling showed that apoptotic index of cardiomyocytes was lower in the cell transplantation group than in the model control group (P

Objective To investigate the effect of baseline Left-ventricular ejection fraction (LVEF) on the cardiac resynchronization therapy (CRT). Methods A retrospective analysis of patients with heart failure was performed. Fifty-four patients [aged (59.73 ± 11.61)years, 31 males, 23 females] who underwent CRT/CRTD were divided into 2 groups according to LVEF ( group A, LVEF≤ 35%; group B, LVEF > 35% ). NYHA class, LVEF,CRT respond rates and medical adverse event (MAE) due to worsening heart failure were detected at 3 and 12 months post-surgery. Results The score of NYHA class changing wasn′t different between these two group. Compared to group B, LVEF was markedly increased (5.08 ± 2.81)% vs (2.45 ± 1.80)%, P < 0.05;(5.38 ± 2.92)% vs (2.39 ± 3.53)%, P < 0.05)at 3 and 12 months post-surgery. Similarly, group A owned high CRT response rates(66.1% vs 33.9%, P < 0.05; 81.25% vs 47.83%, P < 0.05) at the both two time points. At 12 months post-surgery , LVED decreased and the MAE was similar in all groups. Conclusion Patients with EF≤ 35% have more increases in LVEF and own high CRT response rates post-CRT.

Objective To investigate the diagnostic value of urine neutrophil gelatinase apolipoprotein (NGAL) and kidney injury molecule 1 (KIM-1) as markers of CIN,and the effectiveness of hydration therapy in the prevention of CIN.Methods One hundred and twenty patients were randomly divided into control group and treatment group.The patients of treatment group received hydration therapy through intravenous fluid infusion.Urine samples were taken for detecting the value of albumin (mAlb),NGAL,and KIM-1 before surgery (T0),after surgery 12 h (T1),24 h (T2),48 h (T3),72 h (T4) by ELISA assay.The levels of urinary mAlb,Scr,BUN and cystatin C were detected at the same time.Results (1) The urine NGAL/Cr and KIM-1/Cr significantly increased and were more than twice the baseline value at the time of 12 h after PCI in 87 of 120 cases of the participants.There are eight cases occurred CIN (6.67％) and one case occurred in hydration treatment group (1.7％),seven cases were in control group (11.7％).The difference was statistically significant.(2) There were no significant difference in BUN,Scr,mAlb/Cr,Cys-C and GFR between two gToUps (P ＞ 0.05).(3) NGAL/Cr,KIM-1/Cr were elevated at T1 in both groups (P ＜ 0.01).In hydration treatment group,levels of NGAL/Cr and KIM-1/Cr decreased substantially to the level of T0 at T4(P ＜ 0.01),while in the control group they didn't.(4) Area under the ROC curve (AUC) of NGAL/Cr and KIM-1/Cr 12 h after PCI were 0.931 [95％ CI (0.889,0.973)] and 0.811 [95％ CI(0.736,0.886)] respectively (all P＜0.05).Conclusions NGAL and KIM-1 are sensitive and specific indicators for predicting early renal injury induced by contrast medium and can be used for early diagnosis of CIN.Hydration therapy can prevent the contrast agent-induced renal damage.

Objective To investigate influencing factors of perioperative myocardial injury in patients undergone percutaneous coronary intervention (PCI). Methods Ninty one patients with coronary heart disease (including stable angina and unstable angina) underwent PCI, the perioperative myocardial injury incidence were observed prospectively by monitoring the preoperative and postoperative high sensitivity troponin protein levels to investigate the influencing factors of perioperative myocardial injury by Logistic regression analysis. Results There were no statistically significance in perioperative myocardial injury incidence (62.5%vs 68.7%, P=0.618) and perioperative myocardial infarction incidence (29.2%vs 20.9%, P=0.411) between stable angina and unstable angina groups . These factors of perioperative myocardial injury in patients undergone PCI by Logistic regression analysis were analysed, and we found that the influencing factors were the application loading dose rosuvastatin before PCI, preoperative statin therapy more than one month, apolipoprotein A levels and total stent length. Conclusion Application loading dose of rosuvastatin before PCI, preoperative statin therapy more than one month, apolipoprotein A levels are related to perioperative myocardial injury reduction, whereas the total length of the stent is associated with an increased occurrence of perioperative myocardial injury.

This study is to explore the amelioration of piperine on chronic acute combining stress rat with depression-like behavior, visceral sensitivity, and its effect on the expression of serotonin (5-HT) and synaptophysin. Forty two SD rats were divided into seven groups: blank group, model group, piperine (12.5, 25, 50 and 100 mgkg-1, ig) and imipramine (10 mgkg-1, ip) groups. The rat model of irritable bowel syndrome was established by chronic acute combining stress, and then to evaluate depression-like behavior and visceral sensitivity. The expressions of 5-HT and synaptophysin in the hippocampus and colon were determined by high performance liquid chromatography (HPLC) and Western blotting, respectively. The duration of immobility of IBS rat in the forced swimming test had been significantly increased, the sucrose consumption of IBS rat had been reduced and visceral sensitivity was obviously elevated in the IBS model group as compared with those in the normal control group (P<0.05, P<0.01). As compared with those in the normal control group, the expression of 5-HT significantly decreased, 5-HIAA/5-HT ratio significantly increased in the hippocampus of IBS model group (P<0.05), but opposite presentations were noted in the colon (P<0.05). As compared with that in the normal control group, the synaptophysin expression in the hippocampus decreased significantly but obviously increased in the colon (P<0.05). Piperine improved the behavior of IBS rats, and reversed the levels of 5-HT and 5-HIAA, and 5-HIAA/5-HT proportion in the hippocampus and colon (P<0.05); besides, they significantly reverse the synaptophysin level in the hippocampus and colon (P<0.05). The presence of depression and visceral sensitivity had been changed in IBS rats, with abnormal expression of 5-HT and synaptophysin in the brain-gut system. Piperine can ameliorate the changes of the behavior and regulation of serotonin and synaptophysin expression in IBS rat model.

Objective To observe the serum cyclophilin A(CyPA)level change in the patients with coronary heart disease (CHD)and to investigate its clinical significance.Methods 64 patients with CHD(CHD group)were divided into three groups ac-cording to the clinical types:stable angina pectoris(SAP)group,unstable angina pectoris(UAP)group and acute myocardial infarc-tion(AMI)group;which were divided into three groups according to the coronary artery lesion range:single vessel lesion group, double vessels lesions group and multi vessels lesions group;while 26 controls with normal coronary arteries were select as the con-trol group.Serum levels of CyPA and matrix metalloproteinase-9 (MMP-9 )were detected by ELISA,and C-reactive protein(CRP) concentration was detected by the immune scattering turbidimetry test.Results The CyPA,MMP-9 and CRP levels in the CHD group were significantly higher than those in the control group(P<0.01);the CyPA and CRP levels in the AMI group and the UAP group were significantly higher than those in the control group and the SAP group(P<0.05),the CyPA and CRP levels had no statistically significant difference between the SAP group and the control group(P>0.05);serum CyPA,MMP-9 and CRP levels in the single vessel lesion group,the double vessels lesion group,multi vessels lesion group were significantly higher than those in the control group(P<0.05),serum CyPA,MMP-9 and CRP levels were elevated with the increase of coronary artery lesion vessels (P<0.05).In the CHD group,serum CyPA with MMP-9 and CRP showed the significantly positive correlation(r=0.772,0.749, P<0.01).Conclusion Serum CyPA level is significantly increased in the patients with CHD,CyPA may have some relationship to CHD and the plaque stability.

Objective:To study the risk factors for complications after endoscopic retrograde cholangiopancreatography (ERCP) in super-aged patients (≥80 years).Methods:Clinical data of 512 super-aged patients with pancreaticobiliary diseases who underwent 638 ERCP procedures at the Digestive Endoscopy Center, Zhongnan Hospital of Wuhan University, from July 2011 to June 2021, were studied retrospectively. Indications and results of the ERCP operations were analyzed. Multivariate logistic regression model was used to analyze the risk factors for ERCP-related complications.Results:The total success rate of ERCP cannulation in super-aged patients was 94.0% (600/638), which showed no difference compared with that of patients of <60 years old (2 433/2 557, 95.2%) or patients of 60~<80 years old (2 815/3 004, 93.7%) ( χ2=5.49, P=0.064). The overall incidence of post-ERCP complications was 15.2% (97/638), and the in-hospital mortality was 2.1% (11/512), which showed significant difference compared with patients of <60 years old (8/1 809, 0.4%) and patients of 60-<80 years old (21/2 127, 1.0%) ( χ2=13.39, P=0.002). Multivariate regression analysis showed that hypertension ( HR=1.94, 95% CI: 1.237-3.041, P=0.004), history of upper gastrointestinal reconstruction ( HR=2.28, 95% CI: 1.064-4.891, P=0.034), endoscopic sphincterectomy ( HR=1.65, 95%CI: 1.012-2.679, P=0.045), early procedure period ( HR=0.57, 95% CI: 0.352-0.923, P=0.022), operation time >30 minutes ( HR=1.74, 95% CI: 1.094-2.759, P=0.019), preoperative white blood cell count >9.5×10 9/L ( HR=2.66, 95% CI: 1.661-4.257, P<0.001) and procalcitonin ≥0.05 ng/L ( HR=2.54, 95% CI: 1.172-5.513, P=0.018) were independent risk factors for post-ERCP complications. Conclusion:ERCP is safe and effective for super-aged patients. However, much attention should be paid to post-ERCP complications of patients with hypertension, history of upper gastrointestinal reconstruction, endoscopic sphincterectomy, operation time >30 minutes, preoperative white blood cell count >9.5×10 9/L and procalcitonin ≥0.05 ng/L to avoid serious adverse events such as mortality.

Objective:To investigate the correlations of endoscopic evaluation results with laboratory indices and clinical disease activity in Crohn disease (CD) patients with different intestinal involvement.Methods:Data of 147 patients diagnosed as having CD who visited the Department of Gastroenterology, Zhongnan Hospital of Wuhan University from July 1, 2017 to June 30, 2022 were collected retrospectively. According to the involvement of intestinal segment, patients were divided into three groups: the group with isolated small intestinal involvement ( n=55), the group with both small intestinal and large intestinal involvement ( n=48), and the group with isolated large intestinal involvement ( n=44). Correlations of endoscopic evaluation (based on CDEIS) with laboratory indices and clinical disease activity (based on Harvey-Bradshaw index) were analyzed. Results:C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) could be used for the prediction of endoscopic disease activity. The areas under curve (AUC) of receiver operator characteristic (ROC) were 0.677 (0.506-0.849) and 0.744 (0.597-0.890), respectively. In terms of determing clinical disease activity, clinical Harvey-Bradshaw index was consistent with endoscopic CDEIS score in 65.3% (96/147) patients, showing a low positive correlation ( r=0.260, P<0.05). In subgroup analysis for patients with isolated small intestinal involvement, CRP showed no predictive value for clinical disease activity [AUC (95% CI): 0.617 (0.461-0.773), P=0.148], while for endoscopic activity neither CRP nor ESR showed predictive value [AUC (95% CI): 0.537 (0.146-0.929), P=0.829; AUC (95% CI): 0.571 (0.153-0.990), P=0.680]. Furthermore, for patients with isolated small intestinal involvement and both small intestinal and large intestinal involvement, no correlation was found between clinical Harvey-Bradshaw index and endoscopic CDEIS score ( r=0.222, P=0.092; r=0.142, P=0.322). Conclusion:For CD patients with small intestinal involvement, especially isolated small intestinal involvement, laboratory indices and clinical disease activity cannot accurately reflect endoscopic disease activity. Great importance should be attached to evaluation of the extent and activity of intestinal lesions by endoscopy, especially enteroscopy.