BACKGROUND:Tissue and cellimplantation entails high-quality seed cells. In order to satisfy this requirement, it is crucial to produce adequate wel-conditioned, high-purity and strong proliferation ability bone marrow-derived mesenchymal stem cells.
OBJECTIVE:To establish a simple, rapid and effective in vitro isolation and culture method of bone marrow-derived mesenchymal stem cells, and to define the biological features of bone marrow mesenchymal stem cells.
METHODS:Rat bone marrow mesenchymal stem cells were isolated from the bilateral tibial and femoral bones by the method of whole bone marrow, then purified and passaged by attachment method. The morphology and features of bone marrow mesenchymal stem cells were observed, the growth curve was drawn and the cellsurface antigen was detected by flow cytometry. The bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells isolated by the whole bone marrow adherence method grew vigorously and were highly purified. The cultured cells were spindle-shaped. The growth curve was S-shaped and the population doubling time was 29 hours. The cells stil maintained a strong proliferative capacity after they were passaged for 10 generations. The surface markers such as CD44, CD29, CD90 were positive, while CD45, CD34, CD11b were negative. At the third passage, bone marrow mesenchymal stem cells were induced to differentiate along the osteogenic, chondrogenic and adipogenic lineages, respectively. Fol owing induction, Alizarin red staining, alkaline phosphatase staining, von-kossa mineralized nodules staining, toluidine blue staining, and oil red O staining were al positive. This shows that the whole bone marrow adherence method is a simple and reliable method for the in vitro isolation, culture and proliferation of bone marrow mesenchymal stem cells. Moreover, they have multi-lineage differentiation capacity under different inducers. The third passage bone marrow mesenchymal stem cells have the highest biological activity and can act as the ideal seed cells for subsequent experiments.

OBJECTIVE:To evaluate the human body pharmacokinetics and bioequiavailability of two kinds of oral single dose of hydrochloric itraconazole capsules.METHODS:A randomized,crossover study of20healthy volunteers receiving sin-gle oral dose of200mg itraconazole was conducted with in vivo blood concentrations determined by HPLC-fluorescence de-tection.RESULTS:The pharmacokinetic parameters for the testing itraconazole and the reference itraconazole were as fol-lows,t 1/2 were(29.3?5.62)h and(29.3?5.81)h,respectively;C max were(81.4?60.0)?g/L and(77.8?45.2)?g/L,respec-tively;t max were(3.9?0.70)h and(4.2?0.70)h,respectively;AUC 0～72 were(1199.4?649.6)(?g?h)/L and(1174.3?701.9)(?g?h)/L,respectively;AUC 0～∞ were(1414.0?815.2)(?g?h)/L and(1386.1?735.8)(?g?h)/L,respectively.there were no significant differences in main pharmacokinetics parameters between2preparations,except in t max from analysis of variance and one-side&two-sides t-tests.The relative bioavailability of trial itraconazole capsule was(105.3?23.4)%.CONCLUSION:These two kinds of itraconazole capsules are bioequivalent.

Objective To describe the CT and MR imaging features of hepatic inflammatory myofibroblastic tumor(IMT).Methods The CT(n=12)and MRI(n=2)findings of pathologically proved hepatic IMT in 12 patients were retrospectively analyzed.Results All the tumors of the 12 patients were solitary and located in right hepatic lobe.Six tumors were solid and 4 tumors were solid-cystic.The other 2 tumors were periportal soft-tissue infiltration.The tumors appeared as solid or solid-cystic hypodense mass on CT images.The lesions were dark in T1-weighted sequences and slightly bright on T2-weighted sequences.There was homogeneous or inhomogeneous moderate to significant enhancement in solid portion of the tumors.Honeycomb-like enhancement was demontrated both in the peripheral part and at the intratumoral septa of the solid-cystic mass.Conclusion CT and MRI can provide helpful information for the clinical diagnosis and differential diagnosis of hepatic IMT.

Objective To study the effects of cinobufotalin capsule combined with transcatheter arterial chemoembolization in patients with hepatocellular carcinoma.Methods 68 patients with hepatocellular carcinoma from January,2013 to March,2016 were selected and divided into the combination group and chemotherapy group according to the random number table method,each with 34 cases.The combination group was treated with transcatheter arterial chemoembolization and hepatic artery infusion chemotherapy embolization.The levels of D-dimer,tumor markers,clinical efficacy and quality of life scores were observed before and after treatment in 68 patients.Results There was no significant difference in plasma fibrinogen(FIB)in combination group before and after treatment.Before treatment,the levels of FIB)was similar to chemotherapy group.After treatment,the FIB content in the chemotherapy group was significantly higher than before treatment and that in the combination group the differences were statistically significant(P<0.05).After treatment,the levels of cytokeratin 19,carcinoembryonic antigen and neuron-specific enolase in two group were significantly higher than those before treatment(P<0.05).But there was no significant difference between the two groups after treatment.The rate of disease control in the combination group was significantly higher than that in the chemotherapy group(77.5％vs.65.4％,P<0.05).Cases of diarrhea and insomnia in chemotherapy group were similar to the combination group,but the differences of cases of pain and fatigue between two group were statistically significant(P<0.05).Conclusion Cinobufotalin capsule combined with transcatheter arterial chemoembolization can improve the hypercoagulability of the patient's blood,and is beneficial to the treatment of cancer,improve the disease control rate and improve the life of patients.

Objective To evaluate the effectiveness and safety of open tension-free hernioplasty and two kind of laparoscopic hernia repair for the treatment of primary unilateral inguinal hernia. Methods Patients suffering from primary unilateral inguinal hernia were randomly divided into 3 groups to undergoopen operation with mesh-plug and patch or transabdominal preperitoneal hernioplasty (TAPP) or totally extraperitoneal hernioplasty (ZEP). Results From February 2006 to February 2009, a total of 164patients were divided into 3 groups, with 62 undergoing open tension-free mesh-plug hernia repair, 50receiving TAPP and 52 receiving TEP. Postoperative patients were then followed up for a period of(16±8)months. The average operating time for the patients in the open mesh repair group was significantly shorter compared to the other two groups (P＜0.01 ). The cost in the open mesh repair group was also significantly lower than the other two groups (P＜0.01). The pain scores in open mesh group were significantly higher than those in the other two groups (P＜0.01). The hospital stay and the recovery time were both significantly longer in the open mesh repair group (P＜0.01) compared to the other two groups. No major complications and recurrence was found in neither groups. Conclusions Open tension-free mesh-plug hernia repair, TAPP and TEP are all safe and effective for the treatment for patients with primary unilateral inguinal hernia. TAPP and TEP are superior to open tension-free mesh-plug hernia repair as these two procedures involve less postoperative pain and fast recovery.

Objective To investigate the prognostic factors for chronic heart failure and the prognostic ability of copeptin,big endothelin-1(Big ET-1)and N-terminal pro-brain natriuretic peptide (NT-proBNP)in patients with chronic heart failure. Methods To study 1 5 9 consecutive patients hospitalized for chronic heart failure.Serum concentration of copeptin,NT-proBNP,cTnI,CKMB and plasma Big ET-1 as well as left ventricular ejection fraction (LVEF)and NYHA classⅠtoⅣ on admission were measured.Cardiac events were found by patients to discharge after 360~490 days,prospectively.Results During a median follow-up period of 385 days,the endpoint of recurrence for cardiac events was reached in 65 patients with 159 heart failure.Multivarlate canonical correlation analysis shows the older and the higher NYHA classification as well as the lower LVEF in patients with heart failure.There were higher concentration of copeptin,Big ET-1 and NT-proBNP.On a Cox proportional hazards regression models analyses,age,copeptin,Big ET-1 and NT-proBNP were found to be the inde-pendent predictors of cardiac events.Risk ratio (RR)were 1.215,1.236,4.031 and 13.052,respectively.Logistic regression models analyses,copeptin,Big ET-1 and NT-proBNP were found independent predictors of death.Odd ratio (OR)were 4.003,2.477 and 1.235,respectively.Conclusion Measurement of copeptin,Big ET-1 and NT-proBNP in patients with chronic heart failure can help to identify patients at higher risk for cardiac events and patients for prognosis.

Objective: To explore the suitable load of endurance training for primary school students with different BMI levels living in 3 200 m plateau, and to provide guidance for exercise and weight control for children residing in high altitudes. Methods: The heart rate deflection point (HRDP) and HRDPspeed of 7-12 year-old students residing at high altitude were evaluated by using the site Conconi test. The serum lipid metabolism level was detected by ELISA. The correlation between HRDP and lipid metabolism was analyzed by Pearson correlation analysis. Results: The level of leptin, adiponectin, total cholesterol(TC) and triglycerides(TG) in obesity group was highest, follow by the overweight group, normal weight and under-weight group(F=3.75-24.12, P<0.05). In the same age group, hrdpspeed decreased with the increase of BMI. For children with the same BMI classification, HRDPspeed decreased with age. HRDPspeed showed an increasing trend by age and BMI. In obese, normal and emaciated students, there was a significant negative correlation between lipid metabolism and HRDP, but there was no significant correlation between lipid metabolism and HRDP in overweight students of different ages (P>0.05). Conclusion It is suggested that HRDP and lipid metabolism may be signcficantly correlated with BMI in students aged 7-12 years at different BMI levels in plateav and the above indicators can be used as an effective means to evaluate aerobic exercise endurance capacity of healthy adults at plateau.

Objective: To discuss the correlation between aerobic exercise ability, pulmonary function and heart rate recovery ability of 7-12 year-old students under different altitudes, so as to provide evidence on the physiological changes of exercise and characteristics of altitude training among children and adolescents at different altitudes. Methods: A total of 235 students aged 7-12 years old living at 3 240 m(Hi group), 300 students at 1 290 m(SubHi), and 300 students living between 1 290 m and 3 240 m (HiSubHi) were selected to perform 20-mSRT test, respiratory function test and heart rate recovery ability evaluation after quantitative load exercise. Results: Performance on 20-mSRT, FVC, FEV1, FEV1% and V% of students aged 7-12 years at different altitudes showed an upward trend with the increase of age. After adjusting for altitudes, there were significant differences in average level of these indicators. Average level of 20-mSRT, FVC, FEV1%, FEV1% and V% in SubHi group were significantly higher than those in Hi group(P<0.05), and the HiSubHi group showed an upward trend compared with Hi group after 1-year adaptation to the sub-highland environment; Except for male FVC (r=0.33, P=0.24) and FEV1 (r=0.34, P=0.22) in HiSubHi group, and the male FEV1% (r=0.46, P=0.14) in SubHi group, all the indicators showed significant association with 20-mSRT(P<0.05); 20-mSRT was only significantly correlated with heart rate recovery ability among SubHi group (male: r=0.78, P<0.05); Female(r=0.81, P=0.01), there was no significant correlation between 20-mSRT and heart rate recovery ability in other groups(P>0.05). Conclusion The results suggest developmental characteristics of aerobic exercise ability, lung function and heart rate recovery ability among 7-12 years old students under different altitudes. The correlation among 20-mSRT, FVC, FEV1, FEV1% and V% might involve high altitude adaptation.

Objective To evaluate the predictive performance of C2 HEST score in the progression from paroxysmal to persistent atrial fibrillation(AF)in elderly patients.Methods A retrospective cohort trial was conducted on 515 patients＞75 years old with paroxysmal AF admitted to the First and Second Centers of Chinese PLA General Hospital from January 2008 to December 2009.The occurrence of persistent AF(including permanent AF)or death was regarded as the end event.According to their C2 HSET score,the end population was divided into a progressive group(76 cases,progression to persistent AF)and a non-progressive group(439 cases,not progression to persistent AF).The risk factors for persistent AF were analyzed,and the survival rate was ana-lyzed using Kaplan-Meier survival curve.ROC curve was plotted to evaluate the performance of C2 HEST score in predicting the progression to persistent AF.Results Multivariate Cox regres-sion analysis showed that COPD(HR=2.018,95％CI:1.262-3.227,P=0.003),heart failure(HR=3.505,95％CI:1.5 9 8-7.688,P=0.002)and stroke or transient ischemic attack(HR=3.663,95％CI:2.293-5.852,P=0.000)were independent risk factors for progression to persis-tent AF.Kaplan-Meier survival curve analysis indicated that the survival rate was decreased with the increment of risk stratification(P＜0.01).The median survival time of intermediate-risk patients was 124.81 months(95％CI:120.752-128.862)and that of high-risk patients was 97.04 months(95％CI:86.277-107.808).ROC curve analysis revealed that C2 HEST score(AUC value=0.649,95％CI:0.577-0.721,P＜0.01)had good predictive value in the progression of paroxysmal AF to persistent AF.Conclusion In patients over 75 years of age,C2 HEST score has a good performance in evaluating the progression from paroxysmal to persistent AF.

Objective:A mouse model of pancreatitis induced by coxsackievirus B3 (CVB3) was established. The pathological change of pancreas and the infiltration of Th17/Treg cells were observed.Methods:The BALB/c mice were inoculated intraperitoneally with CVB3 to induce acute viral pancreatitis model. Then the pathological changes of pancreas were observed by HE staining; the viral RNA load and relative expression of cytokines (IFN-γ, IL-6 and IL-17) mRNA were detected by q-PCR; the proportion of infiltrated CD45 + CD3 + T cells, CD4 + and CD8 + T cells, Th17 and Treg cells in the pancreas was determined by flow cytometry. Results:Three days after CVB3 infection, the viral RNA load in pancreas was the highest (0.96±0.18) and gradually decreased with prolongation of infection. Compared with the 3 dpi group, the viral RNA load in pancreas was decreased (0.96±0.18 vs. 0.62±0.14) at 7 dpi, but there was no statistically significant difference. In addition, the infiltration of immune cell in pancreas increased significantly after 7dpi and the pathological score >2. The percent of infiltrated Th17 cells (1.05±0.21 vs. 22.13±5.79) and Treg cells (3.11±0.78 vs. 8.25±1.30) among CD4 + T cells significantly increased after infection (P<0.05), and the Th17/Treg also increased (P<0.01). Compared with the control group, the relative mRNA expression of IFN-γ (1.05±0.23 vs. 672.6±47.67), IL-6 (1.00±0.38 vs. 68.28±4.57), and IL-17 (1.01±0.11 vs. 54.15±7.94) in pancreas increased at 7 days after CVB3 infection ( P<0.01). Conclusions:The infiltration of Th17/Treg cells and the expression of related cytokines related cytokines IL-6 and IL-17 mRNA were upregulated in pancreas, which promoted the process of CVB3-induced pancreatitis.