BACKGROUND: Under special conditions, bone marrow mesenchymal stem cells (BMSCs) can differentiate into osteoblasts and chondroblasts. However, MSCs are few in bone marrow. How to harvest, purity and rapidly proliferate in vitro is a foundation of application in tissue engineering technique. OBJECTIVE: To optimize, collect, purity, assess rabbit BMSCs and to observe the biological character of BMSCs. DESIGN, TIME AND SETTING: The observational study was performed at the Animal Experimental Center of Tongji Medical College from September 2005 to July 2006. MATERIALS: One female New Zealand rabbits aged 2 months were used for MSC collection and primary culture. METHODS: Bone marrow solution was purified by density gradient centrifugation and adherence screening method. Culture solution was obtained. BMSCs were incubated in phosphate buffered solution (PBS), supplemented with 2.5 g/L trypsin (3.0 mL), and placed in an incubator at 37 ℃ for two or three minutes. Cell morphology was observed using an inverted microscope. The digestion was stopped when cytoplasm recovery, long and thin cells with large intercellular space, and few round cells appeared. Subsequently, BMSCs were incubated in serum-free L-DMEM, and placed in a plastic culture flask at 1.0×108/L. MAIN OUTCOME MEASURES: MSC morphology, ultrastructura and surface marker; Proliferation of the first, third, fifth, eighth and tenth passages of BMSCs; Cell growth curve was drawn. RESULTS: BMSCs was pure following density gradient centrifugation and adherence screening method. The third and fifth passage of cells had typical whirlpool-shape. Transmission electron microscope demonstrated that round or oval MSCs possessed large nuclei, big nucleus proportion, a few cellular organ. These were low-differentiated cells. Growth curve of cultured MSCs was "S" shape. The first, third and fifth passage cells had strong reproductive capability. The eighth and tenth passage of cells had significantly reduced proliferation. Cells isolated were positive for CD44 and CD90, but negative for CD34. These were low-differentiated cells under the electron microscope. CONCLUSION: Isolated cells are MSCs, with the property of stem cells. The third and fifth passage cells are pure, with strong reproductive capability.

To observe the effects of Huanglian Jiedu Decoction (HJD), a compound traditional Chinese herbal medicine, on lipid metabolism and its related gene expressions in rats with hyperlipidemia.

Objective To explore the clinical features and feasibility of renal transplantation for end-stage renal disease (ESRD) following hematopoietic stem cell transplantation (HSCT).Method A retrospective study was done in one case of renal transplantation for ERSD following HSCT.Clinical manifestations were summarized and prognosis was described.The 22-year-old male recipient had received HLA allele matched related bone marrow transplantation from his sister in 2001 and accepted renal transplantation 14 years after HSCT because of delayed renal dysfunction.Donor was a cardiac death patient,the preoperative Panel Reactive Antibody Testing was negative and there were 1.5 HLA antigen mismatches of 6 HLA-A,B,DR antigens of donor and recipient.The recipient received immunosuppressive therapy of tacrolimus + mycophenolate mofetil + steroid after renal transplantation.Result The patient's renal function remained stable and serum creatinine level was 65 μmol/L.The outcome of the patient was fairly good during the follow-up period of short-term.Conclusion Renal transplant is a feasible alternative for patients with ESRD following HSCT.If the transplanted kidney and abbr.hematopoietic stem cells are from different donors,irnmunosuppressive treatment is essential after renal transplantation.Long-term follow-up and adjustment of immunosuppression treatment are needed to prevent and treat postoperative complications.

Objective To investigate the association between serum betatrophin levels and insulin resistance in newly diagnosed type 2 diabetes patients. Methods A total of 31 healthy subjects (control group) and 35 newly-diagnosed type 2 diabetes patients(new T2DM group) were recruited. Serum betatrophin was detected with the methods of ELISA, insulin resistance was assessed with homeostasis model(HOMA-IR). Results The levels of betatrophin, free fatty acids, HOMA-IR, fasting plasma glucose, fasting serum insulin, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were significantly higher in group B than those in group A (P<0.05). Conclusion Insulin resis-tance leads to elevated serum levels of betatrophin in patients with newly diagnosed type 2 diabetes.

Objective: To analyze the recent postoperative and long-term postoperative complications of open-splenectomy and disconnection in patients with portal hypertension. Methods: There were 1 118 cases with portal hypertension who underwent open splenectomy and azygoportal disconnection from April 2010 to September 2015 at Department of Surgery, People′s Liberation Army 302 Hospital. Retrospective case investigation and telephone follow-up were conducted in October 2016. All patients had history of upper gastrointestinal bleeding before operation. Short-term complications after surgery were recorded including secondary laparotomy of postoperative abdominal hemostasis, severe infection, intake disorders, liver insufficiency, postoperative portal vein thrombosis and perioperative mortality. Long-term data including postoperative upper gastrointestinal rebleeding, postoperative survival rate and incidence of postoperative malignancy were recorded, too. GraphPad Prism 5 software for data survival analysis and charting. Results: Postoperative short-term complications in 1 118 patients included secondary laparotomy of postoperative abdominal hemostasis(1.8%, 21/1 118), severe infection(2.9%, 32/1 118), intake disorders(1.0%, 11/1 118), liver dysfunction (1.6%, 18/1 118), postoperative portal vein thrombosis(47.1%, 526/1 118)and perioperative mortality(0.5%, 5/1 118). After phone call following-up, 942 patients′ long-term data were completed including 1, 3, 5 years postoperative upper gastrointestinal rebleeding rate(4.4%, 12.1%, 17.2%), 1, 3, 5-year postoperative survival rate(97.0%, 93.5%, 90.3%); the incidence of postoperative malignant tumors in 1, 3 and 5 years were 1.7%, 4.4% and 6.2%. Conclusions Reasonable choosing of surgical indications and timing, proper performing the surgery process, effective conducting perioperative management of portal hypertension are directly related to the patient′s short-term prognosis after portal hypertension. Surgical intervention can reduce the rates of patients with upper gastrointestinal rebleeding, improve survival, and do not increase the incidence of malignant tumors.

Objective:To explore the therapeutic effect of low-intensity extracorporeal shock wave (Li-ESW) in model rats with chronic prostatitis and its optimal parameters.Methods:From April to August 2019, 90 healthy male SD rats aged 8 weeks were randomly divided into control group (group C, n=15), model group (group M, n=15) and shock wave treatment group (group T, n=60), which were treated with Li-ESW after being modeled successfully. The rats in group T were supine and treated with focused Li-ESW, and the probe was placed above the anatomical position of the prostate. Treatment scheme was 3 Hz of frequency, 500 pulses, and once a week for 4 weeks. Group T was divided into group T1(0.09 mJ/mm 2), T2(0.20 mJ/mm 2), T3(0.30 mJ/mm 2) and T4(0.40 mJ/mm 2) according to energy flux density, with 15 rats in each group. Before the establishment of the model, the rats in each group were tested with von Frey fiber of 2 g, 4 g and 6 g at the scrotum respectively, and the positive reaction was recorded. There was no significant difference between the three groups ( P>0.05). After one week, the rat model with CP was established by injecting 3% carrageenan into the bilateral lobes of the prostate under anesthesia. And one week after the modeling was completed, the rats in each group underwent von Frey test again to measure pain and evaluate the modeling effects. Group T was then treated once a week for a 4-week period. During the treatment, von Frey test was performed before each treatment to evaluate the treatment effect of the previous week. After the von Frey test for one, two and four weeks of treatment, 5 rats were sacrificed in each group, and the bilateral lobes of the prostate were dissected under aseptic conditions for paraffin inclusion and HE staining. Result:The positive reaction in group M and each group T was significantly more than those in group C ( P<0.001) one week after the modeling was completed, and there was no significant difference between group M and group T. The number of positive reactions in group T2 was lower than that in group M at each time point after treatment ( P<0.05), and it is also less in group T1, T3 and T4 were than that in group M after 3 weeks, 3 and 4 weeks and 2 weeks respectively( P<0.05). In group T2, the number of positive reactions began to decrease after 1 week of treatment, and reached the lowest after 4 weeks, with significant difference in pain measurement results at different time points ( P<0.05). The results of pathological examination showed that there were more inflammatory cells in prostatic stroma and disordered arrangement of epithelial cells in group M than that in group C. The pathological scores of group M, T1, T2, T3 and T4 were 8, 7, 4, 6, 9 after 1 week treatment, 8, 5, 3, 4, 7 points after 2 weeks treatment, and 7, 3, 2, 4, 7 points after 4 weeks treatment, respectively. Conclusions:Low-intensity extracorporeal shock wave treatment had a significant effect on the improvement of symptoms in model rats with chronic prostatitis. Under the scheme of frequency 3 Hz, 500 pulses, once a week for 4 weeks, the optimal parameters is 0.20 mJ/mm 2.

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is one of the common diseases in uroloandrology, which recurs easily after treatment. In recent years, the safety and efficacy of extracorporeal shock wave therapy (ESWT) for CP/CPPS has been widely demonstrated. Studies have shown satisfactory short-term (≤12 weeks) outcomes of ESWT, but lack long-term (>12 weeks) follow-up data. In addition, inconsistent indications and unexplained therapeutic mechanisms have limited the further clinical promotion of ESWT. This article summarizes the latest progress and potential mechanism of ESWT in the treatment of CP/CPPS in order to provide new insights for the standardized application of ESWT.

OBJECTIVE： To study the relationships of polymorphism of MTRR gene rs1801394 locus and SLCO1B1 gene rs11045879 locus with drug concentration of methotrexate （MTX） and high-dose MTX （HD-MTX）-induced ADR in acute lymphoblastic leukemia （ALL） children. METHODS： From Oct. 2015 to Sept. 2018， 70 ALL hospitalized children of Han nationality in Sichuan area who received HD-MTX treatment and were in consolidation chemotherapy were selected retrospectively from Sichuan People’s Hospital. The blood concentration of MTX at 48 and 72 hours after administration was measured by EMIT. The genetic typing of MTRR gene rs1801394 locus and SLCO1B1 gene rs11045879 locus were detected with real-time PCR. The relationships of the polymorphism of MTRR gene and SLCO1B1 gene with MTX blood concentration [dose-corrected concentration （c48 h/D，48 h）， the proportion of children with different concentration of MTX （≤0.1， ＞0.1 μmol/L）] and ADR （such as myelosuppression， liver function damage， gastrointestinal response， mucosal damage， rash， etc.） were analyzed. Binary Logistic regression analysis for the correlation of ADR with different influencing factors （gene polymor- phism， blood concentration of MTX， immunophenotyping， body mass index， etc.） was carried out by Wald method. RESULTS： Totally 31， 32， 7 children with MTRR gene AA， AG and GG genotype， while 23， 37， 10 children with SLCO1B1 gene TT， TC and CC genotype were detected. The distribution of each genotype in 70 children conformed to Hardy-Weinberg equilibrium （P＞0.05）. There was no significant difference in c48 h/D（48 h） of children and the proportion of children with different concentration of MTX （72 h） among difterent genotypes of MTRR and SLCO1B1 gene （P＞0.05）. There was statistical significance in the incidence of liver function injury in children with different genotypes of MTRR gene （P＜0.05）， and the AA genotype was significantly higher than the AG+GG genotype （P＜0.05）. There was no correlation of MTRR gene polymorphism with the incidence of other ADR， neither SLCO1B1 gene polymorphism with the incidence of ADR （P＞0.05）. The results of Binary Logistic regression analysis showed that liver function damage in ALL children was related to the gene polymorphism of MTRR； gastrointestinal reaction was related to whether the plasma concentration was more than 0.1 μmol/L at 72 h； mucosal damage was related to the immune type and BMI of children； the occurrence of skin allergy was correlated with body weight of children（P＜0.05）. CONCLUSIONS： Gene polymorphism of MTRR rs1801394 locus may associated with the occurrence of HD-HTX-induced liver function injury in ALL children， but its polymorphism and gene polymorphism of SLCO1B1 rs11045879 locus are not related to MTX blood concentration in ALL children.