BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.

Objective To explore the association of immune suppression with the severity of HBV-related acute-on-chronic liver failure(ACLF) patients.Methods From August 2009 to April 2010 in Shanghai Public Health Clinical Center, the peripheral blood samples and clinical data of 27 HBV-related ACLF patients (ACLF group), 28 patients wit h chronic active hepatitis B (CHB group)and 8 healthy individuals (Control group) were collected.APACHE Ⅲ score and the grade of hepatic encephalopathy were as quantitative index to evaluate the severity degree of the disease.The absolute counts of the subsets of T lymphocytes and human leukocyte antigen (HLA)-DR expression on the surface of monocyte in patients' peripheral blood were examined by flow cytometry, the proinflammatory cytokines and anti-inflammatory cytokines(IFN-γ, TNF-α, IL-2, IL-4, IL-10) in patients' plasma were detected by cytometric bead array (CBA) kit.The data was analyzed with SPSS 16.0 software.Results Compared with CHB group and control group, the level of anti-inflammatory eytokire IL-10 markedly increased ir HBV-related ACLF patients (Z= -4.279 ,U= 124, P＜0.01;Z= - 3.871, U= 9.5, P= 0.0001 ), however the level of pro-inflammatory cytokines IFN-γ、 TFN-α、IL-2 、 IL-4 in plasma were at low limit of detectable value.Meanwhile the expression quantity of HLA-DR on the peripheral blood monocytes significantly down-regulated (Z= -4.714, U= 98, P＜0.01;7= - 4.086, U = 4, P＜ 0.01), and there was negative correlation between HLA-DR expression quantity and APACHE Ⅲ score (R2 =0.2667, P=0.0167).In addition, the absolute counts of CD4+T lymphocytes in adaptive immune cells significantly decreased (Z= -4.411, U= 116, P＜0.01; Z=-3.575, U= 17, P= 0.0004).Conclusions The immune system of HBV-related ACLF patients displays immune dysfunction like monocyte function inhibition; CD4+ T lymphocytes depletion and high level of anti-inflammatory eytokines, the persistent down-regulation of the HLA-DR expression on monocyte is an indicator for the severity of disease.

Objective To study apoptosis and antigen presentation changes of monocytes in HBV-related acute-on-chronic liver failure (ACLF) patients under immune dysfunction state.Methods Peripheral blood samples of 26 HBV-related ACLF patients (ACLF group),20 active chronic hepatitis B patients (CHB group) and 18 healthy individuals (control group) were collected.The changes of apoptosis and proliferation (Ki67) in monocytes and the expression of surface markers including human leukocyte antigen (HLA)-DR and B7 molecules (CD86) of monocytes were analyzed by flow cytometry. Results The percentage of Annexin V expressed monocytes of ACLF group (64％) was significantly higher than that of CHB group (28％) and control group (20％),and the difference was statistically significant (x2 value was 11.75 and 27.23 ; both P＜0.01),which indicated that monocytes apoptosis increased.The Ki67 expression in monocytes of ACLF group was lower than that of CHB group and control group,and the difference was statistically significant (x2 value was 4.71 and 4.83; both P＜ 0.05),which indicated that activated monocytes reduced. The mean fluorescence intensity (MFI) of HLA-DR and CD86 of monocytes in ACLF group was 22.85 and 11.63,which was significantly lower than that of CHB group and control group,indicating the antigen presentation ability of monocytes injured. The percentage of Annexin Ⅴ positive monocytes in survivals (62 ％ ) was significantly higher than that of dead patients (46 ％ ) in ACLF group.Conclusion In HBV-related ACLF patients under immune dysfunction state,the apoptosis of peripheral blood monocytes increased,and the quantity of activated cells reduced,resulting in the decline of the antigen presentation ability of monocytes.

This paper analyses the data structure of DICOM standard by the applications in the Non-DICOM format fluoroscopic images converted into the DICOM format images. It puts forward a solution to integrate the Multi-Channel Electrophysiology Recorder System with the X -ray system in the cardio-catheter room.
Computer Communication Networks ; Diagnostic Imaging ; instrumentation ; methods ; Electrophysiologic Techniques, Cardiac ; instrumentation ; Fluoroscopy ; instrumentation ; Humans ; Image Processing, Computer-Assisted ; instrumentation ; methods ; standards ; Information Storage and Retrieval ; Radiology Information Systems ; standards ; Software Design ; Video Recording ; instrumentation ; methods ; standards

OBJECTIVETo study the molecular epidemiological characteristics of hantavirus seen during 2000-2003 in Qingdao region of Shandong province.

METHODSSera were collected from 64 patients with hemorrhagic fever with renal syndrome (HFRS) and viral RNA was extracted from the sera. HTN and SEO universal primers were designed as outer primers and HTN and SEO specific primers as inner primers. G1 gene region of M segment from hantavirus was amplified by using RT-nest-PCR for sequencing. The data of nucleotide sequences were analyzed by DNA star software.

RESULTSSix cases were positive by HTN specific primer of total cases (9%); 25 of 64 cases by SEO specific primer (39%); total positive rate was 48%. In general, SEO type was a prevalent type of hantavirus in Qingdao region. The variation of the nucleotide sequences among SEO viruses (nucleotide sequence divergence ranged from 0.3% approximately 8.9%) was lower than that among HTN type (nucleotide sequence divergence ranged from 2.6% approximately 11.2% ).

CONCLUSIONMajority of hantavirus found in Qingdao region belonged to SEO type and still a few strains belonged to HTN type. Most of the HTN viruses were detected in Jiaonan county.

China ; Genotype ; Hantavirus ; genetics ; Hemorrhagic Fever with Renal Syndrome ; blood ; virology ; Humans ; RNA, Viral ; blood ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

Adult ; Cytokines ; metabolism ; Female ; Hepatitis B ; complications ; Humans ; Liver Cirrhosis ; immunology ; metabolism ; pathology ; virology ; Liver Failure ; immunology ; metabolism ; pathology ; Male ; Middle Aged ; Th1 Cells ; metabolism

OBJECTIVETo compare the mid- to long-term outcomes of patients receiving isolated coronary artery bypass grafting (CABG) versus surgical ventricular restoration (SVR) plus CABG for left ventricular aneurysms.

METHODSThe clinical data were retrospectively analyzed in 205 patients with left ventricular aneurysms admitted to our hospital between January, 1997 and December, 2012, including 115 patients receiving SVR plus CABG and 90 undergoing isolated CABG. By matching preoperative echocardiographic parameters including aneurysm size, left ventricular ejection fraction (LVEF), left ventricular end-systolic volume index (LVESVI) and EuroSCORE risk factors, 32 patients receiving SVR plus CABG and another 32 with isolated CABG were enrolled in this study. The patients were compared for survival rates, major adverse cardiac or cerebrovascular events (MACCEs), left ventricular geometry and function at 1, 3 and 5 years of follow-up.

RESULTSCompared with the patients receiving isolated CABG, those receiving SVR and CABG showed greater improvements in echocardiographic parameters and NYHA functional class. The differences in the echocardiographic parameters between the two groups gradually reduced with time and became comparable at 5 years after the operation (P>0.05). No significant difference was found in the mid- to long-term survival or the incidence of MACCEs between the two groups (P>0.05).

CONCLUSIONCompared with isolated CABG, SVR plus CABG does not reduce the incidence of MACCEs or improve the mid- to long-term survival rate of patients with left ventricular aneurysm with a LVESVI <60 mL/m(2).

Aneurysm ; surgery ; Coronary Artery Bypass ; Echocardiography ; Heart Ventricles ; surgery ; Humans ; Incidence ; Retrospective Studies ; Risk Factors ; Stroke Volume ; Survival Rate ; Treatment Outcome ; Ventricular Function, Left

OBJECTIVETo evaluate the expression of TEIF protein in human tumors of soft tissue and its significance.

METHODSAnti-TEIF polyclonal antibody was generated by immunization of E.coli expressed His-TEIF protein. The expression of TEIF in 166 cases of sarcomas and 28 case benign tumors or tumor-like lesions of soft tissue arranged in tissue chip was analyzed by immunohistochemistry.

RESULTSPolyclonal antibody obtained from immunized rabbit was verified in Western blot to prove its specific reactivity with native TEIF protein. The immunohistochemical staining of TEIF showed that about 58% (97/166) of sarcomas were positive and significantly different from that of benign tumors or tumor-like lesions (11%, 3/28). The positive staining was predominantly in synovial sarcoma 94% (16/17), primitive neuroectodermal tumor (PNET) 91% (21/23), both of which were significantly higher than 43% (6/14) of dermatofibrosarcoma protuberans, 38% (6/16) of myxofibrosarcoma, 36% (8/22) of malignant peripheral nerve sheath tumor, 32% (6/19) of liposarcoma, (P < 0.05, respectively), but not higher than 75% (15/20) of malignant fibrous histiocytoma, 70% (7/10) of rhabdomyosarcoma or 64% (9/14) of leiomyosarcoma. Meanwhile, strong positive staining of TEIF (>or= 2+) was frequently observed in PNET (83%, 19/23) and synovial sarcoma (76%, 13/17). With respect to FNCLCC grading, 19 cases of grade I sarcoma TEIF was 32% (6/19) and strong positive was 11% (2/19), 44 cases of grade II sarcoma was 48% (21/44) and 32% (14/44), and 70 of grade III was 84% (59/70) and 70% (49/70). The rate of either positive or strong positive in grade III sarcoma was significantly different from that of either grade I or II (P < 0.05), but no difference between the latter two groups (P > 0.05).

CONCLUSIONSTEIF protein could be detected in large part of soft tissue sarcomas, and it not only over-expressed in most of PNET, synovial sarcomas, but also correlated with histological grading.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; HeLa Cells ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Infant ; Leiomyoma ; metabolism ; pathology ; Male ; Middle Aged ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Rhabdomyosarcoma ; metabolism ; pathology ; Sarcoma ; metabolism ; pathology ; Sarcoma, Synovial ; metabolism ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; Tissue Array Analysis ; Transcription Factors ; biosynthesis ; Young Adult

OBJECTIVETo investigate the potential role of deoxyribonuclease I (DNase I) in the pathogenesis of rheumatoid arthritis (RA).

METHODSDNase I activity was measured by radial enzyme-diffusion method in serum samples from 83 RA patients and 60 healthy volunteers and in the synovial fluid (SF) from 27 RA patients and 38 patients with other inflammatory arthritis. SF cfDNA level was measured with Pico Green Kit, and the correlation among DNase I activity, cfDNA level and clinical parameters of RA patients was analyzed.

RESULTSSerum DNase I activity was significantly lower in RA patients than in the healthy control subjects (0.3065∓0.1436 vs 0.4289∓0.1976 U/mL, P<0.001), and was negatively correlated with ESR (r=-0.2862, P=0.0122), CRP (r=-0.2790, P=0.0184) and neutrophil cell counts (r=-0.287, P=0.011). SF DNase I activity was almost negative in patients with RA, ankylosing spondylitis (AS) and gouty arthritis (GA). SF cfDNA level in RA patients was significantly higher than that in patients with osteoarthritis (100.81∓142.98 vs 18.98∓31.40 µg/mL, P=0.002), but similar to that in patients with AS (45.85∓47.67 µg/mL, P=0.428) and GA (162.95∓97.49 µg/mL, P=0.132). In patients with inflammatory arthritis, SF cfDNA level was positively correlated with ESR (r=0.4106, P=0.0116) and CRP (r=0.5747, P=0.0002).

CONCLUSIONImpairment of DNase I activity may be responsible for the enhanced NETs generation and plays a role in the pathogenesis of RA.

AIMTo explore proper cryopreservative systems for hematopoietic stem cells.

METHODSPeripheral blood mononuclear cells from 20 persons were mixed with different cryopreservative agent, dimethyl suflfoxide (DMSO) or combination of DMSO and hydroxyethyl starch (HES), then cooled in -80 degrees C low temperature refrigerator (Refr) or autocontrolled programmed cryogenic system (PCS), preserved in Refr or in liquid nitrogen. GM-CFU, LTC-IC, CD34+ cells and typeran blue resistance (TBR) were assayed after different period of cryopreservation.

RESULTSThe recovery rates of CFU-GM, LTC-IC, CD34+ cells and TBR in peripheral blood mononuclear cells which were cooled and preserved in Refr with 5% DMSO-6% HES were 82.2% +/- 14.7%, 83.0% +/- 12.2%, 94.2% +/- 4.3% and 97.7% +/- 3.9% respectively, significantly higher than that in Refr with 10% DMSO (P < 0.05). When cells were cryopreservated with the same cryopreservatives, there was no significantly difference of recovery rate in group of Refr and group of Refr with PCS. Meanwhile, there was not significantly difference of recovery rate among all three groups, preserved in Refr ahead of liquid nitrogen, in Refr merely, in liquid nitrogen with PCS within one year (p > 0.05). However, the recovery rate of CFU-GM, LTC- IC, CD34+ cells and TBR decreased dramatically if cells were cooled and preserved in Refr for two years. After cells were thawed, the cell activity declined gradually at room temperature if the cryopreservatives were not removed or diluted. The cell activity of 10% DMSO group was affected more than that of 5% DMSO-6% HES group.

CONCLUSION5% DMSO-6% HES is better than 10% DMSO as cryopreservatives for hematopoietic stem cells. Refr cryopreservation is a simple and effective method if cells would be cryopreserved for less than one year. If cells would be cryopreserved for more than one year, liquid nitrogen cryopreservation should be recommended. The cryopreservatives should be diluted or removed immediately after cells were thawed.

Blood Preservation ; methods ; Cell Survival ; drug effects ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans