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.

Purpose To investigate the expression of mTOR in breast cancer, and to observe the effect of CCI-779 on proliferation and apoptosis of MDA-MB-231 cell. Methods Immunohistochemical staining was used to detect the expression of mTOR protein in breast cancer tissue and MDA-MB-231 cell. MTT method was used to show effect of CCI-779 on proliferation of MDA-MB-231 cell. Annex-inV-FITC/PI method was used to show effect of CCI-779 on apoptosis of MDA-MB-231 cell. Results 54.9% in 71 cases of breast cancer tissue could express mTOR protein, the expression was significantly higher than in 32 cases of normal tissue (21.9%), mTOR protein was also detected in MDA-MB-231 cell, CCI-779 could inhibit the proliferation of MDA-MB-231 cell, and show a dose-and time-dependent, but CCI-779 could not induce apoptosis of MDA-MB-231 with AnnexinV-FITC/PI assay. Conclusion mTOR is closely related to the formation of breast cancer, CCI-779 has strong activity against MDA-MB-231 cell, it has prospect for treatment of breast cancer in the future.

Objective To investigate the influence of high-fat diet on blood pressure and metabolism in Sprague-Dawley(SD) rats and its mechanism.Methods Forty male SD rats were randomly divided into 4 groups which were fed with a diet containing 53% calorie as fat(HF) or a normal diet(ND) for 5 or 10 weeks.Systolic blood pressure(SBP),body weight,abdominal adipose tissue,blood lipids,fasting insulin(FINS),and fasting plasma glucose(FPG) were measured after 5 and 10 weeks respectively.Results SBP of HF groups were higher than that of ND groups [HF5 vs.ND5,(105.506?4.634)mmHg vs.(100.060?4.773)mmHg,P

Objective To explore the effect of beraprost sodium (BPS) on hypoxia-induced pulmonary artery hypertension (HPH) in rats and the expression of oxygen-sensitive Kv channels in pulmonary artery smooth muscle (PASM).Methods The HPH model of rats was established by exposing rats to low-pressure and low-oxygen cabin which was auto-modulated for 8h every day.Rats in the BPS group were given an intragastric administration of BPS [300 μg/(kg · d)],while those in the control group and HPH group were given an intragastric administration of 3 ml/kg of 0.9％ saline.After 4 weeks,the mean pulmonary artery pressure (mPAP) was measured and right heart ventricle hypertrophy index (RVHI) was calculated;pulmonary artery remodeling was evaluated by HE staining;the expressions of Kv 1.2,Kv 1.5 and Kv2.1 in the pulmonary artery were detected by Real-time PCR and Western blot.Results The HPH model was successfully established in rats exposed to chronic hypoxia for 4 weeks.Compared with those in HPH group,mPAP,RVHI and pulmonary artery remodeling were decreased in BPS group [mPAP:(13.48±2.18)mmHg vs.(23.87±2.23)mmHg vs.(17.09±1.20)mmHg;RVHI:0.28±0.02 vs.0.46±0.03 vs.0.36±0.04;％ area of medial smooth muscle:35.72±6.58 vs.68.52±5.64 vs.46.58±8.43;P＜0.05],and the mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 were increased (relative protein expression level:Kv1.2,0.78±0.10 vs.0.15±0.03 vs.0.57±0.13;Kv1.5,0.61±0.10 vs.0.31±0.05 vs.0.59±0.13;Kv2.1,0.29±0.05 vs.0.10±0.02 vs.0.28±0.07;P＜0.05).Conclusion BPS can improve pulmonary arterial hypertension induced by hypoxia,and upregulate the decreased mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 in pulmonary artery.

Objective To examine the change of body weight (BW) and blood pressure (BP) in obese rats, clarify relationships between BP and BW and other factors. Methods Male Spraque-Dawley rats were fed either with normal diet (ND) or high calorie diet (HC) for 20 weeks. BW and BP of tail artery were observed biweekly and tetraweekly respectively; serum leptin and fasting insulin (FINS) were detected by enzyme-linked immunoadsordent assay (ELISA) and radioimmunoassay (RIA) respectively. Fasting plasma glucose (FPG) and free fatty acid(FFA) were measured by conventional means. Results BW, abdominal fat weight (AFW), ratio of abdominal fat weight to body weight (RF/W), systolic blood pressure (SBP), diastolic blood pressure (DBP), serum levels of leptin and FINS, FPG, FFA increased in the HD group after 20 weeks diet intervention (P＜0.05 or P＜0.01). SBP was strongly correlated with BW, leptin, FINS and FFA (P＜0.05), DBP was correlated with FFA (r=0.47, P＜0.05). In addition, leptin was positively correlated with BW, AFW, RF/W, FINS and FFA (P＜0.05 or P＜0.01). Conclusion In this study of high calorie-diet induced rats, the gain of BW is accompanied by increased BP. The obese rats have hyperleptinemia, hyperinsulinemia, hyperglycemia and dyslipidemia which may have important effects on the development of obesity-related hypertension. RF/W is the key factor in which affect serum leptin level.

Objective To assess whether there was strong association between antiphospholipid antibodies(APA) and coronary heart disease(CHD), to study the environmental factors of APA production and APA pathogenic mechanism in patients with CHD.Methods Blood samples from 76 patients with CHD and 30 controls were tested for anticardiolipin antibodies IgG(ACA-IgG),human cytomegalovirus IgG,IgM(HCMV-IgG,IgM) by enzyme-link immunosorbant assay(ELISA) and 6-keto-PGF1a,endothelin(ET) by radioimmunoassay(RIA).Results A total of 27 patients(35.53%) were ACA positive in 76, as compared to 2 of 30(6.67%) healthy individuals, P＜0.05. There was no difference in ACA among acute myocardial infarction(AMI,39.13%), old myocardial infarction(OMI,26.53%), unstable angina pectoris(UA,38.40%), P＞0.05. The number of ACA positive subjects was higher in HCMV infection patients with CHD than no HCMV infectious patients with CHD. There was no PGI2 and ET level difference between ACA-IgG positive and negative CHD.Conclusion There are strong association between APA and CHD. The HCMV infection may be an environmental factor of APA production in CHD patients with raised ACA. The alteration of PGI2 and ET are not the pathogenic mechanism of ACA in patients with CHD.

OBJECTIVE:To study the pharmacokinetic characteristics of Puerarin solid self-microemulsion capsules in rats in vi-vo. METHODS:The rats were randomly divided into two groups with 18 rats in each group. They were given Puerarin solid self-mi-croemulsion capsules(S-SMEDDS capsules)and commercially available Yufeng ningxin tablet,with the dose of 158.15 mg/kg puer-arin. The 0.2 ml blood samples were collected from tail vein 0,5,10,20,40,60,90,120,180,240,360,480,600 min after medication,respectively. The blood concentration of puerarin was determined by HPLC,and pharmacokinetic parameters and rela-tive bioavailability were calculated by using 3p87 software. RESULTS:The metabolism of puerarin was one-compartment model in rats. Pharmacokinetic parameters of S-SMEDDS capsules and Yufeng ningxin tablet were as follows as cmax of (1.032 0 ± 0.020 6) and(0.587 3±0.011 7)μg/ml,t1/2ke of(116.431 4±2.166 0)and(88.222 6±1.752 4)min,AUC0-t of(261.532 2±1.464 0)and (102.835 5±1.957 4)μg·min/ml. Relative bioavailability was 238.77%. CONCLUSIONS:Compared with Yufeng ningxin tablet, S-SMEDDS capsules are absorbed more completely and removed faster.

OBJECTIVE:To provide reference for rational utilization of antibiotics for special use. METHODS:Medical orders of antibiotics for special use in clinical departments of our hospital during 2013 to 2015 were selected from hospital information sys-tem. The consumption sum and its ratio,DDDs,DDC,DUI and the utilization of antibiotics for special use in clinical departments were calculated and analyzed. RESULTS:The consumption sum and its ratio of antibiotics for special use in our hospital during 2013 to 2015 both reduced year by year,decreasing from 21 872 200 yuan(48.00%)to 20 877 700 yuan(39.41%). The consump-tion sum ratio of carbapenems,cephalosporins and anti-deep fungal drugs showed descending tendency. The consumption sum ratio of anti-MRSA antibiotics changed slightly. DDDs of Meropenem for injection always took up the first place in recent 3 years,but the values were decreasing. DUI of Imipenem and cilastain sodium for injection was far less than 1,while those of Panipenem and betamipron for injection,Cefepime sodium for injection and Caspofungin acetate for injection(70 mg/injection)were far more than 1. The types of drug with DUI ranged 0.9-1.1 increased year by year,increasing from 3 types in 2013 (18.75%) to 10 types in 2015 (62.50%). Within 3 years, the utilization ratio of antibiotics for special use always took up the first place in ICU (86.64%-87.78%). CONCLUSIONS:The consumption sum and its ratio of antibiotics for special use in our hospital during 2013 to 2015 decreased year by year,and the utilization of antibiotics for special use become increasingly rational. But,there still are some problems,such as inadequate dose of Imipenem and cilastain sodium for injection,overdose of Panipenem and betamipron for injection,Cefepime sodium for injection and Caspofungin acetate for injection(70 mg/injection).

Objective To explore the correlation of helicobacter pylori(HP) infection in liver cirrhosis(LC) of different syndrome patterns with blood ammonia level.Methods Two hundred and seventeen LC patients were divided into Hp-positive group and Hp-negative group.The patients were classified into 6 syndrome patterns: liver-Qi stagnation,internal accumulation of water-dampness,stagnation of damp-heat,deficiency of liver and kidney yin,deficiency of spleen and kidney yang,and blood-stasis blocking collaterals.Parameters of blood ammonia level,hepatic function and blood coagulation function were observed in LC patients with different syndrome patterns.Results The Hp infection rate in 217 LC patients was 40.1%,the difference being insignificant in the patients with different syndrome patterns and hepatic function grading.The percentage was in a decreasing sequence in syndrome patterns of stagnation of damp-heat,internal accumulation of water-dampness,blood-stasis blocking collaterals,deficiency of liver and kidney yin,liver-Qi stagnation,deficiency of spleen and kidney yang.The difference of blood ammonia level was significant between Hp-positive group and Hp-negative group(P0.05).Conclusion Hp infection is an important factor of inducing the increase of blood ammonia level in LC.Blood ammonia level increases in LC patients after Hp infection,especially in patients with blood-stasis blocking collaterals.The hepatic function grading is corrected with blood ammonia level,the worse the hepatic function,the higher the blood ammonia level.