BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.

Background and purpose: High-dose chemotherapy followed by autologous stem cell transplantation (auto-HSCT) is considered as the ifrst line treatment for patients with relapse/refractory lymphoma after conventional chemotherapy. However, most of these patients still relapse the second time. The purpose of this study was to analyze the efifcacy of the consolidation chemotherapy after autologous stem cell transplantation (HSCT) refractory/relapse lymphoma in high risk. Methods:A total of 38 patients with relapsed/refractory lymphoma including Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) were included, who were underwent auto-HSCT in our transplan-tation department from Jan. 2010 to Dec. 2013. In treatment group, 19 patients received 2 courses of consolidation che-motherapy after auto-HSCT every 2 to 3 months, with the regimen of mini-BEAM or modiifed mini-CBV. Another 19 patients had no chemotherapy after auto-HSCT as control group. Results:The median follow-up duration was 17.2 and 7.5 months in the treatment and control group respectively. The follow-up data demonstrated prolonged progression-free survival (PFS) in the treatment group than the control group [24.7 months vs 7.8 months, P=0.029 under intend-to-treat analysis ITT;24.7 months vs 5.2 months, P=0.01 under per protocol analysis(pp)]. There is also a trend of improved overall survival (OS) in the treatment group (P=0.055, ITT). Conclusion:Consolidation chemotherapy after auto-HSCT for refractory/relapsed lymphoma patients delay the relapse and tend to improve the overall relapse rate.

Objective To evaluate the therapeutic efficacy of transcather arterial chemoembolization for hepatic carcinoma and its influence on hepatic function. Methods Patients with HCC were divided into three groups according to repeated TACE times. The therapeutic efficacy was assessed with survival rate, car-cionma size and biologic response (AFP). Hepatic function was evaluated according to some hepatic param-eters(ALT,AST,ALB,DB,TB,PT). Results All three groups responses rates were 4.3%, 23.1% and 31.6% ;srespectively,liver function in group 1 and group 2 returned to its pretreatment level(P <0.05) ,but in group 3, liver function did not return to its pretreatment level associated with more repeated TACE times. The 1-,2-and 3-year survival rates of group 1 were 27.0%, 10.8%, 5.4% ; respectively, while those of group 2 were 57.1%, 33.3%, 19.0% ; respectively, which were not significantly different from those of Team 3 ,which was 62.5%, 37.5%, 18.8%, respectively. Conclusions (1) three to four repeated times TACE was an effective palliative treatment that prolongs survival of patients with HCC,while 1 to 2 re-peated times had a limited benefit; (2) Irreversible hepatic impairment induced by TAEC could affect its therapeutic efficacy.

Objective To investigate the treatment options of acute biliary pancreatitis.Methods A retrospective review was performed in 134 consecutive patients who presented to a single tertiary care institution from 2000 to 2010 with acute biliary pancreatitis.Results Of the 134 patients,30 had a persistent (commonbile duct,CBD)stone.Following multivariate analysis,at admission CBD size on ultrasound,alkaline phosphatase(ALP),total bilirubin(TB),significantly correlated with persistent CBD stone.Receiver operator curve analysis and linear regression were applied to obtain optimal and equitable predictive values,and variables combined.Optimal values were.,CBD≥10 mm;AP≥150 U/L;and TB≥51.3μmol/L.Presence of three variables had an associated odds ratio(OR)of29.5(P<0.001)for presence of persistent CBD stone.Zero variables conferred asignificantly decreased probability of CBD stone,OR 0.07(P<0.001).Conclusions Presence of three variables significantly correlated with persistent CBD stone.Biliary evaluation by endoscopic retrograde cholangiopancreatography(ERCP)is suggested.In the absence of any positive predictive variables,cholecystectomy may be sufficient.Decisions regarding patients with one to two be made vailables should occur on a case-to-case basis.

Objective To study the regulation effect of estrogen in expression of matrix metalloproteinase-9 (MMP-9) in the central nervous system (CNS) in mice with experimental autoimmune encephalomyelitis ( EAE).Methods The 60 mice were overiectomized and 2 weeks later EAE was induced with MOG35-55 peptide in these mice.They were divided into a treatment group and a control group.The treatment group was treated with estrogen and the control group was given PBS.Clinical symptoms in these two groups were scored and compared.HE staining was used to observe inflammation in the brain and spinal cord.The MMP-9 expression in the CNS was examined by quantitative real-time PCR and immunofluorescence staining.Results The incidence of disease was lower (treatment and control group were 8/30 and 28/30 respectively) and clinical symptoms were milder (treatment and control group were 3.23±0.83 and 1.62 ±1.00 respectively,t=3.811 and P＜0.05) in the treatment group than those in the control group.HE staining showed the decreased infiltration of inflammatory cell in the treatment group (Treatment group:inflammatory score were 0.895 ±0.206,0.752 ±0.302,0.732 ±0.183 in acute,relief and chronic phase respectively;Control group:inflammatory score were 3.472 ±0.635,2.881 ±0.662,1.891 ± 0.482 in acute,relief and chronic phase respectively.t = 8.622,6.543 and 5.027,all P ＜ 0.05).The quantitative real-time PCR and immunofluorescence staining showed that the expression of MMP9 in the CNS was decreased in the treatment group.Conclusion Estrogen may decrease MMP-9 expression in the CNS,reduce inflammation and clinical symptoms in mice with EAE.

ObjectiveTo explore the clinical effect of video-assisted thoracoscopic surgery(VATS) for chronic empyema. Methods74 patients with chronic empyema were divided into experimental group and control group,control group was given decorticationoflung,and experimental group was given VATS. ResultsThe duration of chest tube drainage, hospitalization and complications in experimental group was significantly better than that in control group,it showed better clinical effect. ConclusionFor part of patients without the serious disease change of chronic empyema, if the surgery indication could be controlled strictly, the VATS was considerable.

Objective To explore the methods of thoracoscopic intrapleural perfusion hyperthermic chemotherapy(TIPHC)on diagnosing and treating malignant pleural effusion caused by lung cancer,as well as its effect.Methods From February 1999 to March 2006,seventy patients with malignant pleural effusion caused by lung cancer were randomly divided into therapeutic group(35 cases)and control group(35 cases).Pleural biopsy and TIPHC under general anesthesia with unilateral ventilation were performed in the therapeutic group,and intrapleural injection of cisplatin was administered in the control group after drainage of pleural effusion.The effect on malignant pleural effusion,the change for the concentration of carcino-embryonic antigen(CEA),cytokeratin-19 fragments(CYFRA21-1),neuron-specific enolase(NSE)and the side effect were compared before and after the treatment.Results The therapeutic group achieved total response rate of 100%,but only 54.3% in the control group,with significant difference(P

Objective To explore the clinical characteristics,diagnosis,treatment,and prognosis of male breast cancer.Methods The clinical data of 38 cases of male breast cancer,who were treated in the two hospital during the past 10 years,were analyzed retrospectively.The diagnosis was mainly based on preoperative B-ultrasound,radiography with molybdenum target tube and fine needle aspiration.Among the 38 cases,radical mastectomy was conducted in 3 cases,modified radical mastectomy in 27 cases,simple mastectomy in 5 cases,and operation was refused in 3 cases.Postoperatively,radiotherapy was adopted in 4 cases,simple endocrinotherapy in 7 cases,radiotherapy plus chemotherapy in 9 cases,and chemotherapy plus endocrinotherapy in 8 cases.Results Median age at treatment was 58.5(53-82)years.TNM staging included stageⅠin 13 cases,stageⅡ in 19 cases,stageⅢ in 4 cases,and stageⅣ in 2 cases.Median follow-up was 73.8 months(2 months-10 years),and follow-up rate was 73.6%(28/38).During the follow-up,13 cases died including 8 cases died of local recurrence and metastasis,and 5 cases died of heart and brain disease.The 5-and 10-year overall survival rate was 71.6% and 55.3% respectively;5-and 10-year disease-specific survival of stageⅠ,Ⅱ was 92.6% and 78.9% respeoively;among 6 cases of stageⅢ,Ⅳ 4 were followed,and all 4 cases died with in 3 years.Conclusions Male breast cancer has a long course,poor prognosis and low survival rate.Combined therapy with modified radical mastectomy,as its basis,is the treatment of choice for male breast cancer.There is a certain correlation between the prognosis of male breast cancer and clinical stage of the disease.

Objective To assess the impact of HLA-A,HLA-B,HLA-DRB1 matching on the prognosis of hematopoietic stem cell transplantation from unrelated donors.Methods A total of 81 patients with hematological malignancies including leukemia,myelodysplastic syndrome(MDS)and lymphoma who underwent hematopoietic stem cell transplantation from unrelated donors from 2007 to 2012 in our department were included in this retrospective analysis.Patients were classified into HLA match group(n=53)and HLA mismatch group(n=28)according to the HLA high-resolution matching.The overall survival (OS),treatmentrelated mortality(TRM),relapse rate(RR),graft-versus-host disease(GVHD)incidence were analyzed.Results The 81 patients were analyzed with a median follow-up of 11.9 months(0.3 to 57.4 months).The OS (66.0％vs 46.4％,P=0.031)and TRM(17.0％vs 42.9％,P=0.017)were significantly different between the HLA match group and HLA mismatch group,while the RR had no significant difference(14.3％vs 32.1％,P=0.111).Multivariate analysis showed HLA matching was an independent prognostic factor of TRM,but not OS.There's no significant difference of aGVHD(22.9％vs 40.9％,P=0.122)and cGVHD (40.0％vs 46.7％,P=0.655)incidence between the two groups,but the incidence of severe aGVHD in HLA match group were much lower(4.2％vs 25.0％,P=0.005)than HLA mismatch group.Conclusion the high-resolution matching of HLA-A,-B,DRB1 affect OS,TRM and the incidence of severe aGVHD in unrelated hematopoietic stem cell transplantation,but not affect RR,the incidence of aGVHD and cGVHD.

Objective To observe the influence of Qingwen Baidu decoction (QBD) on serum procalcitonin (PCT) and C-reactive protein (CRP) levels in septic rats and study the mechanism of heat-clearing and detoxifying method for treatment of sepsis. Methods Fifty male Sprague-Dawley (SD) rats were randomly divided into control group (n=5), model group (n=25), and heat-clearing and detoxifying experimental group (experimental group, n=20). The septic model was reproduced by intra-peritoneal injection of lipopolysaccharide (LPS, 5 mg/kg). In control group, an equal volume of normal saline was given. After modeling for 2 hours, the heat-clearing and detoxifying experimental group received QBD the first time (composition of the decoction: Gypsum Fibrosum Recens 30 g, Rehmanniae Radix 10 g, Bubali Cornu 15 g, Coptidis Rhizoma 4 g, Gardeniae Fructus 5 g, Platycodonis Radix 5 g, Scutellariae Radix 5 g, Anemarrhenae Rhizoma 5 g, Paeoniae Radix Rubra 5 g, Scrophulariae Radix 5 g, Forsythiae Fructus 5 g, Glycyrrhizae Radix 5 g, Moutan Cortex 5 g, Lophatheri Herba 5 g) by gavage (0.01 mL/g); the rest administration time was 08:00 to 09:00, once a day. The rats in model group were given an equal volume of warm water by gavage. At different time points after modeling, the blood of 5 rats in control group, model group, and experimental group was collected from the abdominal aorta. The serum PCT and CRP levels were tested by the enzyme linked immunosorbent assay (ELISA), and the pathological changes in lung and intestinal tissue were observed under a light microscope. Results Compared with the control group, the PCT level of the model group after modeling for 2 hours was significantly increased (ng/L:332.32±22.85 vs. 70.46±3.18, P<0.01), this situation continued until 72 hours after modeling. CRP level in the comparison between the control and model groups did not have statistical significant difference at 2, 8, and 48 hours after modeling (all P > 0.05). The level of CRP in model group was lower than that of control group at 24 hours and 72 hours after modeling (μg/L:281.34±32.81, 237.84±41.42 vs. 350.09±56.67, P<0.05 and P<0.01). There were no statistically significant differences in PCT levels between model group and experimental group after modeling for 8 hours and 24 hours (both P > 0.05). The PCT level of experimental group was significantly lower than that of model group beginning from 48 hours after modeling (ng/L: 321.57±28.00 vs. 358.12±10.14, P < 0.05), and this situation continued until 72 hours after modeling (ng/L: 269.50±49.10 vs. 347.69±26.90, P <0.05). The CRP level of experimental group was significantly lower than that of model group beginning from 8 hours after modeling (μg/L:232.73±13.29 vs. 335.35±53.78, P<0.05), this statistical significant difference between the two groups persisted until 72 hours after modeling (μg/L:177.31±6.70 vs. 237.84±41.42, P<0.05). Compared to those in the model group, the lung tissue inflammatory cell infiltration, the intestinal mucosal inflammation and interstitial edema were milder in the experimental group. Conclusion Heat-clearing and detoxifying therapy can effectively reduce the serum PCT and CRP levels of septic rats induced by LPS, and it can alleviate the infiltration of inflammatory cells in lung tissues so as to play a role in protection of tissue organ.