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

From March 2004 to March 2005, 43 patients (49 hips) admitted to Department of Orthopedic Surgery, Fushun Central Hospital for minimally invasive total hip arthroplasty (THA) were selected, including 22 male and 21 female aged 22-79 years. All patients underwent THA with cementless prosthesis. The preoperative Harris scores were (46.2?5.3). For comparison, 35 patients (43 hips) were selected undergoing standard THA at the same period, including 22 male and 13 female, aged 31-78 years; 16 patients underwent THA with mixed prosthesis (cementless prosthesis for acetabulum; cemented prosthesis for the femur), and the others with cementless prosthesis. Their preoperative Harris scores were (43.4?4.6). The perioperative bleeding, operation time, incision length, postoperative functional recovery, and prosthetic position were compared. The THA was succeeded in all patients, and all subjects were followed up for 13.1 months averagely. The perioperative bleeding, drainage, blood transfusion and incision length in minimally invasive THA group were significantly less than standard group (P

The enzyme-inhibitor model and the sugar tolerance mouse model were used to evaluate the relationship between the inhibition rate of enzyme activity and concentration of Hippophae rhamnoides L. subsp. chinensis Rousi polysaccharide (HRP). The inhibitory patterns of enzyme and dose-dependent effects of HRP's effect on blood glucose using acarbose tablets as control were also examined. The mechanism underlying hypoglycemic effects of HRP was discussed. The results showed: in the enzyme-inhibitor model, the inhibitory activity of different concentrations of HRP (9.80, 19.60, 39.20, 78.40, 156.80 and 312.50 mg x L(-1)) on alpha-glucosaminidase (AG) inhibitory activity were 6.62%, 18.02%, 33.26%, 48.23%, 62.11%, 76.31%, 90.12%, IC50 was 31.59 mg x L(-1). The inhibitory rate of 25.00 x 10(3) mg x L(-1) acarbose tablets was only 64.87%, and IC50 was 10.75 x 10(3) mg x L(-1). In the sugar tolerance mouse model, different doses of HRP (240, 480, 960 mg x kg(-1)) tended to decrease levels of blood glucose compared with control group (acarbose tablets 375 mg x kg(-1)) at 15, 30, 60 and 120 min. It's further confirmed that HRP is a kind of competitive inhibitor of AG activity. Its inhibition rate increases with the increase of concentration in normal mice, and it subsequently improves the sugar tolerance showing the effect of reducing blood sugar.

Objective To study the clinicopathological significance of the expression of glutameta decarboxylase 65(GDA65) and protein kinase C(PKC) in the central cancer tissues, cancer edge tissues, paracancerous liver tissue and non-cancer liver tissues. Methods The expression of GDA65 and PKC were detected by immunohistochemical method in 10% neutral formalin- fixed and routinely paraffin-embedded sections in 37 hepatic cancer specimen. Results The positive rate and the score of GDA65 and PKC in the cancer tissues were significantly higher than that in the paracancer tissues or non-cancer liver tissues, but the PKC expression was no difference between the central cancer tissues and the cancer edge tissues . The expression of GDA 65 was related to the pathological types, differentiated degrees, liver cirrhosis or metastasis of hepatocarcinomas. No correlation was found between the expression of PKC and the clinicopathological features of hepatocarcinomas. Conclusions The expression of GDA65 and PKC might be closely related to the carcinogenesis of hepatocarcinoma, they might be important biological markers of hepatocarcinoma.

Objective To analyze the etiology and clinical features of drug-induced liver damage,in order to draw more attention to this problem. Methods Two hundred and fifty-six cases over a 10-year period were retrospectively analyzed. Results A variety of drugs,in-cluding traditional Chinese medicines(23.8% of our total cases)and immunosuppressive agent(11.7%)caused liver damage. The main clinical manifestations of it were fatigue,nausea,vomiting and jaundice. In 78.5% of our cases,the symptoms were relieved or completely disappeared, but there was still 6.6% mortality rate. Conclusion The clinical features of drug-induced liver damage are of no specificity,and the mortality of it is high. Liver function should he monitored when suspected are prescribed.

Objective To study the effect of expression of myoglobin which mediated by adenovirus,on ATP value of liver and the protective effect on liver ischemia reperfusion injury.Methods Adenovirus carrying CMV promoter sequences linked to the human myoglobin gene(AdCMVMyo) were transfected into rats liver. Then myoglobin, hepatic ATP levels and liver function were evaluated. Results Myoglobin expression was verified in rat livers after AdCMVMyo transfection. The ATP levels in rat livers 72 hours after AdCMVMyo transfection were significantly higher than that in control group(P

To explore the feasibility and efficacy of isolated caudate lobe resection for caudate lobe in huge hepatocellular carcinoma(10 cm or larger in diameter).
 Methods: Clinical data of 27 patients with hepatocellular carcinoma larger than 10 cm who underwent isolated caudate lobe resection from January 2001 to December 2011 were retrospectively analyzed.
 Results: All the patients successfully completed the operation. There was no postoperative death. Median operative time was 288 min, and the estimated intraoperative blood loss was 2 260 mL. Postoperative morbidity rate was 44.4%. The patients were discharged successfully after active treatment. Overall survival rates at 1, 3, and 5 years were 80.2%, 52.1%, and 27.1%, respectively.
 Conclusion: Isolated caudate lobe resection is safe and effective for caudate lobe huge hepatocellular carcinoma.
Carcinoma, Hepatocellular ; mortality ; surgery ; Hepatectomy ; Humans ; Liver Neoplasms ; mortality ; surgery ; Retrospective Studies ; Survival Analysis ; Treatment Outcome