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.

A chiral high-performance liquid chromatography method was developed for the simultaneous determination of ibuprofen enantiomers in dog plasma. It was used to study the pharmacokinetics in the Beagle dog after intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen. Ketoprofen was chosen as the internal standard. After a simple precipitation using methanol as the precipitating solvent, both analytes and IS were separated on a Kromasil 100-5CHI-TBB chiral column (250 mm x4.6 mm, 5 μm) with isocratic elution using acetonitrile - 20 mmol x L(-1) phosphate buffer (pH 3.0, containing 5% methanol) (6 : 4) as the mobile phase. The detection wavelength was 220 nm. Liner calibration curves for both of the ibuprofen enantiomers were over the concentration range from 0.5 to 50 μg x mL(-1) with a lower limit of quantification of 0.5 μg x mL(-1), the accuracies were all in standard ranges. The intra- and inter- assay precisions were all below 7%. The recovery rate was 93.1% to 100.4%. The experiments proved that the method was simple, rapid and sensitive. It can be used in the quantitative determination of ibuprofen enantiomers in dog plasma. The method was used to determine the concentration of ibuprofen enantiomers in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen (9 mg x kg(-1)) and the pharmacokinetics parameters were calculated based on the concentration-time curves. The C(max) of S-ibuprofen in Beagle dog plasma after a single intravenous administration of racemic-ibuprofen, S-ibuprofen and R-ibuprofen were 30.8 ± 4.7, 46.1 ± 5.9 and 20.0 ± 2.6 μg x mL(-1), respectively. In terms of the exposure of active ingredient, it revealed a significant difference between the administration of S-ibuprofen and the other two groups. The systematical R- to S- chiral inversion was discussed. Comparing the pharmacokinetic parameters at different doses, chiral inversion were 70.1% ± 36.6% and 76.4% ± 36.2%, respectively, after intravenous administration of racemic- and R-ibuprofen. This study provides a theoretical basis for the safety of ibuprofen formula of injection drug.

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 .

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 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.

Objective:To investigate the effect of continuous blood purification (CBP) on inflammatory factors and immune function in patients with sepsis,in order to provide reference for clinical treatment of sepsis.Methods:A total of 76 patients with severe sepsis,who were treated in ICU of Daqing Oilfield General Hospital from June 2014 to December 2016,were selected.The patients were divided into conventional group (n=36) and CBP group (n=40) according to the received treatment method.The patients in the conventional group were treated with targeted therapy and life support,while the patients in the CBP group were treated with CBP on the basis of targeted therapy.The levels of interleukin-6 (IL-6),interleukin-10 (IL-10),tumor necrosis factor-α (TNF-α) and C reactive protein (CRP) of the patients in the two groups were detected and compared before treatment and after 3d of treatment.The CD3+,CD4+,CD8+T cell and the CD4+/CD8+ ratio before treatment and after 3 d of treatment were calculated and compared between the two groups.Results:There was no significant difference in the level of serum inflammatory factors between the two groups before treatment (P＞0.05).The levels of serum inflammatory factors in the two groups were significantly decreased after 3 d of treatment,and the levels of IL-6,TNF-α and CRP in the CBP group were significantly lower than those in the conventional group (P＜0.05).There was no significant difference in immune function indexes between the conventional group and the CBP group before treatment (P＞0.05).After 3d of treatment,CD3+,CD4+ and CD4+/CD8+ ratio were significantly increased in the two groups,and the CD4+ and CD4+/CD8+ ratio of the CBP group were sig-nificantly higher than those of the conventional group (P＜0.05).Conclusion:The application of CBP in the treatment of patients with sep-sis can obviously improve the expression of inflammatory factors of patients,and can improve the immune function and promote the re-covery of immune function.