BACKGROUND:At present,there are two main methods of isolating and purifying bone marrow mesenchymal stem cells(BMSCs):density gradient centrifugation and the whole bone marrow adherent method.The former has complicated procedures,and the latter has simple operation,but the purified outcomes are not ideal.OBJECTIVE:To establish a rat BMSCs isolated and cultured in vitro purification methods on the basis of the whole bone marrowadherence and isolation of BMSCs in combination of differential passage digestion method.METHODS:By whole bone marrow adherent culturing to isolate and differential digestion and passage of rat BMSCs,the speed ofMSCs in the process of digestion and passage was quicker than other bone marrow cells,as well as the characteristics of adherentspeed,instead of density gradient centrifugation to separate and purify MSCs,and their morphological characteristics wereobserved.Cell growth and proliferation of two kinds of culture method were compared with the density gradient centdfugationseparation.Alkaline phosphatase and oil red staining results were observed to verify BMSC differentiation capacity,to detect thecall surface markers,to validate immune properties and to test its purity.RESULTS AND CONCLUSION:The whole bone marrow adherent culture method isolated and differentially subcultured ratBMSCs.Flow cytometry and osteogenic and adipogenic culture results displayed cytoirnmunity property,purity and differentialcapacity,which have no significant difference compared with density gradient centrifugation.However,cell viability andreproductive activity are obviously elevated.

Objective To explore molecular mechanisms underlying the in vitro counteracting effect of curcumin on malignant melanoma.Methods Cultured A375 and C8161 human melanoma cells were cultivated in vitro,and randomly divided into several test groups and a control group to be treated with different concentrations of curcumin and dimethyl sulfoxide respectively for different durations.Then,methyl thiazolyl tetrazolium (MTT) assay,Transwell assay,flow cytometry and Western blot were performed to evaluate the effect of curcumin on the proliferation,invasion and cell cycle of,as well as expressions of AKT/mTOR signaling pathway-related proteins in A375 and C8161 cells respectively.Statistical analysis was carried out by using t test.Results MTT assay showed that the treatment with curcumin of 5-35 mg/L for 24-96 hours significantly inhibited the proliferation of both A375 and C8161 cells compared with that with dimethyl sulfoxide (all P ＜ 0.001),and the inhibitory effect was in a dose-dependent manner within the range of 5-15 mg/L for A375 cells and within the range of 5-10 mg/L for C8161 cells,and in a time-dependent manner from 0 to 48 hours for both cells.After treatment for 24 hours,the 50％ inhibitory concentration (IC50) of curcumin against A375 cells and C 8161 cells was 10 mg/L and 5 mg/L respectively.Transwell assay demonstrated that the invasion of A375 and C8161 cells was significantly suppressed by 72-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively (both P ＜ 0.001).Flow cytometry showed that the cell cycle of A375 and C8161 cells was arrested at G2/M phase after 24-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively,with significant differences in the proportion of A375 cells and C8161 cells in G2/M phase between the test group and control group (A375 cells:35.00％ ± 3.54％ vs.120.80％ ± 7.46％,P＜ 0.001;C8161 cells:19.33％ ± 4.04％ vs.85.00％ ± 9.53％,P ＜ 0.001).Western blot revealed that the expressions of AKT/mTOR signaling pathway-related proteins were decreased in A375 and C8161 cells after 24-hour treatment with 10 mg/L and 5 mg/L curcumin respectively.Conclusion Curcumin can inhibit the proliferation and invasion of A375 and C8161 cells,likely by blocking cell cycle and inhibiting activation of the AKT/mTOR signaling pathway.

Objective To observe the changes of the heart rate turbulence(HRT)in patients with isolated systolic hypertension(ISH).Methods 85 patients with ISH,80 patients with common hypertension and 58 control subjects were investigated.Turbulence onset(TO)and turbulence slope(TS)were recorded of 24 hours dynamic electrocardiogram.Results The difference of HRT in the three groups was significant(P＜0.01).The values of TO and TS was significantly higher in common hypertension group and ISH group than in control group(P＜0.05);TS was significantly lower in ISH group than in common hypertension group(P＜0.05),but the difference of TO between ISH group with common hypertension group was not obvious.Conclusion The HRT in patients with primary hypertension is significantly lower,which is more significant in patients with ISH than in patients with common hypertension.It suggests that the cardiac autonomic nerves dysfunction perhaps has an important influence on development of hypertension and its prognosis.

Objective To examine the levels of serum adipunectin (ADPN)in patients with end-stage renal disease (ESRD), and identify the relationship between serum ADPN and tumor necrosis factor (TNF)-α, high sensitivity C-reactive protein(hs-CRP) and renal function. Methods Sixty ESRD patients and twenty normal healthy persons were involved in this study. The patients were divided into three groups: patients with chronic renal failure (CRF, 18 cases), patients with hemodialysis (HD,22 cases) and patients with peritoneal dialysis (PD,20 cases). The levels of serum ADPN, TNF-α and hs-CRP were detected by means of ELISA. Differences in continuous variables be-tween the three groups were evaluated. Results The levels of serum ADPN, TNF-α and hs-CRP in CRF, HD and PD patients were signifi-cantly higher than those in control group ( P<0.01 ). Serum ADPN levels in ESRD patients were positive related to serum TNF-α( r≥0.478, P<0.01 ), and negative related to GFR ( r≥-0.582, P<0.01 ). Conclusion The level of serum ADPN was markedly in-creased in patients with ESRD, and correlated with serum TNF-α level and.renal function.

Objective To investigate the levels of serum interleukin(IL)-18 in patients with chronic renal failure (CRF)and its relationship with relevant indexes. Methods Sixty patients with CRF were divided into three groups: patients with CRF (CRF group, n = 18) ,patients with bemedialysis (HD group, n = 22) and patients with peritoneal dialysis (PD group,n = 20). 20 healthy subjects were taken as controls. The levels of serum IL-18, tumor necrosis factor (TNF)-α and high sensitivity C-reactive protein (hs-CRP) were detected by means of ELISA. Serum albumin (ALB) ,total cholesterol (TC) ,triglyceride (TG) and serum creatinine (SCr) levels were detected by con-ventional means. The GFR was measured by using the abbreviated Modification of Diet in Renal Disease study (MDRD) equation. Differences in continuous variables between the three groups were evaluated. Results The lev-els of serum IL-18 were significantly increased in CRF[(497.7 ± 120.7) ng/L], HD[(538.1 ± 113.2) ng/L] and PD[(565.7 ± 122.1) ng/L] patients than those in healthy subjects[(163.9±42.2) ng/L] (P <0.01). Further-more,serum IL-18 levels in HD patients were higher than these in CRF patients (P <0.05). Serum IL-18 levels in CRF were positively related to serum TNF-α,hs-CRP and SCr(r =0. 636,0.436,0.367,P <0.01) ,and negatively related to GFR(r = -0.515 ,P <0.05). Conclusion The levels of serum IL-18 are markedly increased in patients with CRF,HD and PD,and related to the levels of TNF-α,hs-CRP and GFR,so IL-18 might be an important marker of sub-clinical inflammation in patients with chronic renal failure.

Objective To probe the clinical application of CT in the diagnosis of criss-cross heart(CCH).Methods Five patients with CCH confirmed by operation were retrospectively analyzed.Enhanced 64-slice spiral CT was performed in 2 patients and enhanced single-slice electron beam CT was performed in 3 patients.Three dimensional reconstructions were applied for the fully display of anatomic malformations,and the results were compared with that of echocardiogram and angiocardiogram with Chi-square test Results(1)Visceroatrial situs solitus,twisted and concordant atrioventricular connection,horizontally oriented ventricular septum,ventricular septum defect and pulmonary stenosis were found in all patients on CT.The ventriculoarterial connection was discordant,including double-outlet right ventricle in 4 patients and complete transposition of great arteries in 1 patient In addition,associated anomalies including persistent left superior vena cava(n=2),coarctation of the aorta(n=1)and right aortic arch with right descending aorta(n=1)were detected as well.(2)Total 33 anomalies in 5 cases were found during operation.The diagnostic accuracy of CT,angiocardiogram and echocardiogram was 93.9%(31/33),81.8%(27/33)and 54.5%(18/33)respectively.There was a significant difference between CT and echocardiogram(X~2=13.39,P＜0.01),and no significant difference between CT and angiocardiogram(X~2=1.29,P＞0.05).Conclusion CT is an excellent imaging technique for the diagnosis of CCH.

Objective To investigate the clinical application of electron-beam CT(EBCT)in the diagnosis of congenital cardiovascular diverticula. Methotis Retrospective analysis of 9 patients with congenital cardiovascular diverticula confirmed by operation and pathology was done.Of them,enhanced continuous volume scan was performed on 8 patients and enhanced single slice scan was performed on one patient with an Imatron C-150 scanner.Results The group of 9 pailents included one patient with diverticulum of the left ventricle.3 patients with diverticulum of the atria and 5 patients with diverticulum of the aorta.EBCT scan and three dimensional reconstruction could demonstrate not only the origin,size,shape,Location and adjacent structure of diverticula,but also other important complicated abnormalities such as ventrieuloarterial connection disorder,cardiac sepud defect,aortic coarctation and even dissection.Conclusion EBCT is an ideal noninvasive technique in the diagnosis of congenital cardiovascular diverticula

Objective To explore the effect of constitution nursing care by Traditional Chinese Medicine (TCM) theory on urinary salt scaling in patients with ureteral stents.Methods Thirty patients with ureteral stents after ureteroscopy were recruited in the study.The TCM constitution Scale was used to perform the TCM constitution categorization.Then constitution nursing by TCM theory was done based on the symptoms and diagnosis.The urinary salt scaling in the patients was explored. Result In the 30 patients,the incidence of urinary salt scaling were 26.7% (8/30)and no urinary tract infection occurred. Conclusion The nursing by TCM theory can relieve the clinical symptoms by indwelling ureteral stents and reduce the incidence of urinary salt scaling.

Brain metastases are common in oncologic patients. Stereotactic radiosurgery (SRS) is an effective treatment for oligo brain metastases. However, SRS is not suitable for large metastatic lesions. Fractionated stereotactic radiation therapy (FSRT) is a new option for the treatment of brain metastases. It can deliver high doses of radiation to tumor while protecting the normal tissues and organs outside the irradiation field as much as possible and maintaining good local control rate of tumors. This article reviews the dose-response relationship and adverse reactions of FSRT with different fractionated dose schemes, and the progress of FSRT combined with targeted therapy and immunotherapy.

Objective: To explore the clinical features and complications of foreign bodies in the upper gastrointestinal tract in children and to investigate the effectiveness of endoscopic management. Methods: Data of patients with foreign bodies in upper gastrointestinal tract were collected retrospectively at Endoscopy Center, the Children's Hospital, Zhejiang University School of Medicine, from January 2011 to December 2016. Clinical characteristics, the types of foreign bodies, the location and duration of foreign body impaction were summarized. The risk factors of complications and endoscopic removal failure were analyzed by using Logistic regression analysis. Results: A total of 1 334 patients (825 males and 509 females) were enrolled. The median age was 2.5 years, with a range from 0.25 to 15 years and peak age 1-3 years. Twenty patients had esophageal diseases. The most common foreign body ingested was coin (n=964, 72.3%). Foreign bodies were most commonly located in the esophagus (n=1 002, 75.1%), especially in the upper esophagus (n=857, 85.5%). The duration of foreign body impaction ranged from 3 hours to 5 years. Among 1 334 patients, 252 patients (18.9%) developed complications, including ulcers (n=101, 40.0%) and perforations(n=13, 5.2%). The success rate of endoscopic removal was 96.6% (n=1 288). By Logistic regression analysis, sharp foreign body ingestion (OR=6.893, 95%CI: 4.421-10.746) , esophageal impaction (OR=5.253, 95%CI:3.352-8.233) and foreign body impaction longer than 24 hours (OR=4.336, 95%CI:3.091-6.082) were risk factors of complications. Sharp foreign body ingestion was the risk factor of endoscopic failure (OR=5.372, 95%CI:2.773-10.406) . Conclusions Coin is the most common foreign body in upper gastrointestinal tract. Sharp foreign bodies impacted in the esophagus over 24 hours increase the risk of complications. Endoscopic removal of foreign bodies from the upper gastrointestinal tract in children has a high success rate. Sharp foreign body ingestion increases the risk of failure in endoscopic removing.