BACKGROUND:Cord tissues,which immune system is relatively non-development with low expression of MHC,rely on the protection of the mother's immune system.Accordingly,studies underlying biological characteristics of umbilical cord mesenchymal stem cells (MSCs) arouse more and more attention.OBJECTIVE:To isolate,separate,and culture MSCs from the tissues of umbilical cord,and to explore the ability of multi-directional differentiation.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2006 to May 2007.MATERIALS:The umbilical cord of healthy neonate with 37-40 weeks gestational age was obtained from delivery room of the Second West China Hospital,Sichuan University.METHODS:Pieces of umbilical cord were dissected along its long axis,vessel was pulled away,then cord was tied together with a suture creating "loops",then it was placed into collagenase solution,after 6-8 hours,suspended cells obtained from the loops suspension were centrifuged and cultured with adherent method,pass,aged when single cell-derived colonies were appeared.Finally,the 5th generation of cells was induced differentiation towards osteoblasts and lipoblasts.MAIN OUTCOME MEASURES:Morphology,growth,surface antigen expression,as well as the multiple differentiation potential of umbilical cord MSCs.RESULTS:The adherent cells obtained from umbilical cord after removal vessels showed short rod-like or fusiform shape,which can be proliferated and form single cell-derived colonies.Cells isolated from the umbilical cord high expressed integrin markers (CD29,CD51,CD71) but not hematopoietic lineage markers (CD34,CD45 and HLA-DR).These colonies can differentiate into osteoblasts that produce mineralized matrices,stained by alizarin red and alkaline phosphatase,and differentiate adipocytes that accumulate lipid vacuoles and are demonstrated by morphology,stained by oil red.CONCLUSION:Umbilical cord tissue contains a high number of MSC-like elements forming colonies that may be successfully expanded in culture.The proliferation colonies present matrix ceil immunophenotypes,and can differentiate into osteobiasts and adipocytes.

ObjectiveThe purpose of this research was to investigate functional connectivity of 16-channel electroencephalograph(EEG) in δ frequency band based on Granger causality analysis.MethodsThe experimental data was recorded at a sampling rate of 200 Hz from temporal lobe epilepsy (TLE) patients(6 left and 3 right TLE,and 9 normals as control group.Ten of EEG segments of 20 s length for three different states:epileptiform discharges (ED) state in interictal durations,non-ED state for TLE patients,and control state for the normal.The δ band components (1～4 Hz) were filtered from EEGs.The functional connection values Iδ between two EEG δ components were calculated separately by Granger causality analysis.The two EEG components were from inferoposterior temporal lobe (left:T5,right:T6) to frontal lobe (Fp1,Fp2,F3,F4,and parietal lobe (C3,C4) for three states.ResultsThe Iδ values for ED state was 0.1323±0.0329～0.1670±0.028 9,which was significantly higher than that of non-ED state (0.0300±0.0130～0.0420±0.0072) (P＜0.05).The Iδ values for the control group (0.0153±0.0028～0.0193±0.0057) was much lower than that of ED state (P＜0.01),and no obvious distinctions were observed compared with non-ED state at P=0.05 level.ConclusionThere is a stronger connection of EEG' s δ bands from the inferoposterior temporal lobe to frontal and parietal lobe for the ED state,and the over-discharges transmission is from inferoposterior temporal lobe to other brain regions.There is a weaker connection from the top temporal lobe to frontal and parietal lobe for non-ED state and control group,and the onset zones is not inferoposterior temporal lobe.

Objective To explore the relationship between osteopontin (OPN),its downstream product thrombin-cleaved osteopontin (NTOPN) and radiographic changes in osteoarthritis (OA).Methods The levels of OPN and NTOPN in serum and synovial fluid were tested by enzyme-linked immuno sorbent assay (ELISA) kit in 22 OA patients and 21 health controls.The correlation between the above parameters and clinical data such as radiology changes of OA patients was analyzed.Results The serum level of OPN in the OA group was positively correlated with C reactive protein (CRP) (r =0.574,P =0.005).The serum level of OPN in the OA group was slightly lower than that in the healthy control group [(1.8 ± 0.5) μg/L vs (1.7 ± 1.1) μg/L,P ＞ 0.05].Similarly,the serum level of NTOPN in the OA group was significantly higher than that in control group [(21.1 ± 11.7) μg/L vs (14.7 ± 3.6) μg/L,P ＜ 0.05].Meanwhile,serum NTOPN level in female patients was significantly higher than that in male patients [(23.2 ± 11.80) μg/L vs (11.1 ± 4.9) μg/L,P ＜ 0.05].In synovial fluid of OA group,the level of OPN was (7.75 ± 2.33) μg/L,NTOPN was (0.91 ± 0.49) μg/L and the ratio of NTOPN/OPN was 0.12 ± 0.08.The level of OPN of synovial fluid in the OA group was positively correlated with CRP (r =0.574,P =0.005).NTOPN-to-OPN ratio of synovial fluid in the OA group was positively correlated with Kellgren-Lawrence grade (r =0.535,P =0.022).Conclusions The serum level of OPN is correlated with severity of inflammation in the OA patients.Meanwhile,NTOPN-to-OPN ratio of synovial fluid is correlated with OA severity.As a downstream bioactive product of OPN,NTOPN may play an important role in OA pathogenesis.Whether these parameters can be used as predictors of joint lesion severity and inflammatory status is worth further study.

Objective To study the impacts of medicine and pharmacy separation reform on medical insurance fund and medical care seeking behavior of such patients.Methods The insurance data of patients was collected from year 201 1 to year 2014 at the hospital of the author.The differences of hospitals' revenue from insurance,the use of insurance fund and medical care seeking behavior of patients,before and after the reform were analyzed.Results The results showed that the effect of medicine and pharmacy separation reform in achieving translation;the proportions of drug and per-patient drug expenses were reduced.However,the demand of insurance fund was increased,and the fund used in outpatients began to exceed that used in inpatients after the reform.Totally,the choices of outpatients on doctor's technical titles were influenced by the pricing,as more outpatients tended to choose the intermediate title doctors,and the proportion was increasing.The differences of the choices on the outpatient in superior departments were very small,while those in non-dominant departments were very big,the same as the total patients.Conclusion There are both advantages and disadvantages in impacts of the reform on medical insurance fund conducted in some hospital in Beijing.The advantages included that the hospitals'revenue was not decreased from insurance patients,the use of insurance on paying drugs were reduced by management of proper medication, and out-of-pocket burden of the insurance patients was reduced.Yet the disadvantages contained that because of cheaper drugs and favorable insurance policy on the reform,the outpatients preferred higher level hospitals to buy drugs only,and the reform might lead to the increase of the requirement of insurance fund.

Objective To evaluate reliability and validity the Chinese-version of chronic pain acceptance questionnaire-8(CPAQ-8).Methods The reliability and validity of CPAQ-8 was tested in 223 old patients with chronic pain.Result The Chinese-version of CPAQ-8 was (23.79±-5.23),AE was (12.84±4.96),PW was(10.97 ±2.90).Cronbach' s of CPAQ-8 was 0.853,and Guttmann Split-Half Coefficient was 0.843.Two factors were extracted by factor analysis through exploratory factor analysis,whose cumulative contribution was 58.217％.While confirmatory factor analysis showed probability level of the modified model was P=0.322,with indexes as follows: x2/df =1.124,GFI =0.979,AGFI =0,956,TLI =0.069,RMSEA =0.024,NFI =0.960,TLI =0.992,CFI =0.995.Conclusion Reliability and validity of CPAQ-8 were both well.

Objective To analyze the differences of adverse drug reactions between children and adults caused by Tanreqing injection,and provide reasonable application of Tanreqing injection.Methods Retrospective research method were adopted to screen out the adverse drug reaction reports caused by Tanreqing injection from drug ADR reports of Drug ADR Monitoring Center affiliated to the Second People's Hospital of Gansu Province from January 2010 to October 2012.Figures of children and adults who used Tanreqing injection were analyzed separately.Results A total of 603 children used Tanreqing injection,among which 5 children appeared ADR (0.83％).A total of 4395 adult used of Tanreqing injection,among which 4 appeared in ADR (0.09％).There was statistical difference between the two groups (x2 =16.07,P＜ 0.05) The incidence rate of ADR in males was more than females either in the adults or in children.Hypersensitivity was the most common ADR.Conclusion ADR of Tanreqing injection in Children was significantly higher than adults.Tanreqing injection should be used carefully and rationally.

Aim To study the role and mechanism of opioid receptor on IL-1?-induced fever.Methods Rats were dealed with intracerebreventricular administration of naloxone and/or IL-1?.Changes in body temperature were measured.The content of cAMP and HSP70 expression in hypothalamus were detected.Results Naloxone can limit IL-1?-induced fever and decrease the content of cAMP and HSP70 expression in hypothalamus(P

Objective:To develop HMGA1-silencing SMMC-7721 cell and investigate its impacts on apoptosis and cell cycle in tumor cells.Methods:Constructing HMGA1-silencing SMMC-7721 cell through G418 selection of RNAi plasmid transfected cells;surveying the apoptosis,prliferation and cell cycle of the tumor cells by FACS and MTT.Results:Stable HMGA1-silencing cells were obtained successfully.The apoptosis percentage in RNAi group(29.46?3.04%) was significantly higher than that in the negative control (1.96?0.76%) or control (2.04?0.70%)within each P

Objective:To establish a mouse fibroblastic cell line stably transfected with HMGA1 gene,and to use the cell line to investigate the role of HMGA1 in tumor development and progression.Methods:Eukaryotic expression vector pcDNA3.0-HMGA1 was transfected into NIH3T3 by lipofectamine-mediation.Stable transfectants were selected by G418.The expression of extraneous HMGA1 gene was analyzed by RT-PCR and DNA sequencing.Cell growth was measured through MTT and the cell cycle by FCM.Soft agar colony formation was employed to analyze the ability of anchorage-independent growth.The expression of the immune inhibition factors of VEGF and FasL mRNA was detected by RT-PCR.Results:NIH3T3 cell lines stably expressing HMGA1 gene were successfully established.Comparing with controls,the HMGA1 gene-transfected cells grew faster,acquired the ability of anchorage-independent growth and expressed the immune inhibition factors of VEGF and FasL mRNA.Conclusion:Extraneous expression of HMGA1 gene can induce malignant transformation of NIH3T3 and modulate mRNA expression of immune inhibition factors.HMGA1 gene plays a key role in tumor development,progression and immune escape.

Objective To observe the change of regulatory T cells expression in severely burned rats gut ,To investigate the effects of regulatory T cells on CD3+CD4+ /CD3+CD8+lymphocytes and its relation with gut-origin endotoxin translocation .Meth-ods Fifty SD male rats were randomly divided into normal control group(n=10)and burn model groups(n=40) .Rats were burned to achieve Ⅲ degree scalding ,and a 30% total body surface area(TBSA) burn model was made .Rats were sacrificed before(normal control group) and after 0 .5 ,1 ,2 ,4 burn hour(PBH groups) .Flow cytometry techniques were used for the detection of the expres-sions of regulatory T cells and CD3+CD4+ /CD3+CD8+ lymphocyte in intestinal lymph nodes which were separated .The dynamic turbidity method was used for detection of endotoxin levels in portal vein blood .Results The expression of regulatory T cells was negativelycorrelatedwithCD3+CD4+ /CD3+CD8+ lymphocyteratio(r= -0.827,P<0.01)inintestinallymphnodeofrats,while the regulatory T cells was positively correlated with ET levels in portal vein blood plasma .(r=0 .782 ,P<0 .01) .Conclusion The expression of the regulatory T cells in intestinal lymph node in severely burned rats was increased compared to that in normal con-trol group .Regulatory T cells suppressed the expression of intestinal T lymphocytes ,leading to gut immune inhibition .The translo-cation of intestinal endotoxin has a close relationship with regulatory T cells in severely burned rats .Regulatory T cells could have portal effects on intestinal immunity barrier .