BACKGROUND: Endothelial colony-forming cells (ECFCs) transplantation exerts beneficial impact on angiogenesis of impaired tissues caused by ischemia. However, whether this impact is related to the paracrine effect of ECFCs needs to be studied further.OBJECTIVE: To detect the cytokine secretion profile of human umbilical cord blood derived-ECFCs conditioned medium (ECFCs-CM) and explore the effect of ECFCs-CM on the proliferation, migration and tube formation ability of human umbilical vein endothelial cells (HUVECs).METHODS: Human cord blood derived-ECFCs were isolated, cultured in vitro, and then identified based on the previous studies. The cytokines in serum free ECFCs-CM were detected using a cytokines antibody array. HUVECs were cultured with ECFCs-CM, serum free EBM-2 as control. The proliferation, migration and tube formation abilities of HUVECs were examined by cell counting kit-8, scratch test and Matrigel assay, respectively.RESULTS AND CONCLUSION: The cultured cells demonstrated typical characteristics of ECFCs, which showed cobblestone appearance and were positive for CD34, KDR and CD144, but not CD45 or CD133, uptook Dil-acLDL and bond FITC-UEA-I, and tube-like structures were formed on Matrigel. The cytokine antibody array showed that ECFCs-CM significantly upregulated the expressions of 30 kinds of angiogenic factors. Compared with the control group,the HUVECs cultured with ECFCs-CM showed significantly improved proliferation ability at 24, 48 and 72 hours (P <0.01). The migration rate of HUVECs in the experimental group was significantly higher than that in the control group at 12 and 24 hours (P < 0.01). There were more tubular structures in the experimental group than those in the control group (P < 0.05). These results indicate that ECFCs can promote the bioactivity of mature endothelial cells through paracrine action.

Acute Disease ; Cohort Studies ; Humans ; Leukemia ; diagnosis ; Myelodysplastic Syndromes ; diagnosis ; Prognosis

Objective To investigate the correlation between the CYP2E1 enzymes and alcoholic liver disease.Methods Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)method was used to determine polymorphism of CYP2E1.A case control study of 183 subjects was carried out including 40 cases of alcoholic liver disease,40 alcohol dependent group,40 nonalcoholic liver disease and 63 controls.Results The frequency of B genotype or C2 allele gene was significantly higher in alcoholic liver disease than in control(P

OBJECTIVE:To investigate the effect of Saccharomyces boulardii adjuvant with aluminium phosphate on symp-toms and signs disappearance time,immune function and inflammatory factors in children with acute diarrhea. METHODS:Totally 110 children with acute diarrhea were chosen from our hospital during Mar. 2013-Mar. 2016,and then randomly divided into con-trol group and observation group according to random number table,with 55 cases in each group. Control group was given Alumini-um phosphate gel 2.5 g,po,qd,on the basis of conventional treatment. Observation group was additionally given S. boulardii cap-sules 0.25 g,po,bid,on the basis of control group. Both groups were treated for 3 d. Clinical efficacies,symptoms and signs(di-arrhea,abdominal pain,vomiting,abnormal stool routine)disappearance time as well as stool frequency,stool character scores, immune function indexes(CD3+,CD4+,CD8+,CD4+/CD8+),inflammatory factors(IL-6,IL-10,TNF-α)before and after treat-ment,the occurrence of ADR were compared between 2 groups. RESULTS:The total response rate of observation group (90.91%)was significantly higher than that of control group(70.91%);the symptoms and signs disappearance time was signifi-cantly shorter than control group,with statistical significance(P<0.05). Before treatment,there was no statistical significance in stool frequency,stool character scores,immune function indexes or inflammatory factor levels between 2 groups(P>0.05). After treatment,stool frequency,stool character scores and inflammatory factor levels of 2 groups were significantly lower than before, and the observation group was significantly lower than the control group;the levels of CD3+,CD4+,CD4+/CD8+ were significantly higher than before treatment and control group,and the level of CD8 + was significantly lower than before treatment and control group,with statistical significance(P<0.05). There was no significant difference in immune function indexes of control group be-fore and after treatment(P>0.05). Total incidence of dry mouth and constipation was 5.45% in control group and 9.09% in obser-vation group,without statistical significance(P>0.05). CONCLUSIONS:S. boulardii adjuvant with aluminium phosphate in the treatment of pediatric acute diarrhea can efficiently improve symptoms and signs,shorten disease duration,improve immune func-tion and reduce inflammatory reaction with good safety.

Objective To evaluate the effect of contrast-enhanced ultrasound(CEUS) in the real-time monitoring treatment of ultrasound-guided high intensity focused ultrasound (HIFU ) for uterine adenomyosis .Methods From January 2013 to September 2013 ,a total of 67 patients who were clinically diagnosticated as uterine adenomyosis were enrolled in this study .49 patients were diffuse type and 18 patients were localized .CEUS was used before ,in ,after ultrasonic treatment to make sure lesions and evaluate the ablation effect .Results 67 patients were 100% perfused before HIFU treatment by CEUS ,the non-perfused volume ratio of 15 patients(22 .4% ) were not enough during treatment ,and more energy were given .During HIFU treatment ,uniform grey scale change was observed in 42 of 67(62 .7% ) patients and massive grey scale change was observed in 25 of 67(37 .3% ) patients .The margin of targets and the non-perfused volume were clearly detected by CEUS .The size of target andymyosis ,the volume ,the non-perfused volume ,and non-perfused volume ratio were(4 .8 ± 1 .5)cm ,(63 .1 ± 57 .2)cm3 ,(59 .1 ± 53 .4 )cm3 ,(76 .4 ± 27 .0)% .The adverse effects occurred in the treatment were not related to CEUS .Conclusion CEUS is an accurate method in assessing the treat-ment effect of HIFU and could be safely used during HIFU treatment for uterine adenomyosis .

Objective To establish a reliable approach for quantification of colony forming unit(CFU)of Mycobac-terium tuberculosis (M.tb)by measuring optical density(OD).Methods M.tb suspension H37Ra was prepared using low-power ultrasonic or glass bead beating methods,and was two-fold serially diluted,OD at 600nm (OD600)of each dilution ratio was measured respectively,OD600 and dilution curve were analyzed to determine the optimum approach for preparing bacterial suspension,linear range of OD600,as well as linear relationship between OD600 and CFU.Results OD600 was 0.1 -0.6,linear regression analysis of OD600 and dilution ratio within linear range revealed that correlation coefficient (R2 )of glass bead beating and low-power ultrasonic methods were 0.98 and 1 .00 respectively,both presented a good correlation,low-power ultrasonic method was better than glass bead beat-ing method,bacterial suspension dispersed more evenly.Linear regression analysis results of OD600 and CFU val-ues showed that the regression equation of glass bead beating method and low-power ultrasonic method were CFU=2.35×107 ×OD600+4.42×105 and CFU=3.26×107 ×OD600+6.89×105 respectively.Conclusion Low-power ultrasonic method is a good method for preparation of M.tb suspension,combined the measurement of OD600 value, it can be a reliable and rapid method for quantitative analysis of M.tb.

Objective: To clone and express the preferentially expressed antigen of melanoma (PRAME) gene in E.coli. Methods: The cDNA encoding human PRAME gene was extracted from human AML cell line HL-60 and amplified by RT-PCR, then the PRAME gene was inserted into plasmid PGEM-T-easy. After sequenced, it was cloned into the prokaryotic expression vector pGEX-4T-2 to construct a clone with high level expression and the recombinant plasmid was cloned in E.coli. Results:The protein product reached the highest level at 5 h after IPTG induction. Conclusion: Since PRAME is only expressed at low levels in a few normal tissues and encodes an antigen recognized by autologous cytotoxic T lymphocytes, while expressed at high levels in patients with leukemia, it might be a new targeting candidate for tumor immunotherapy.

Objective To examine the effect of endothelial colony-forming cells conditioned medium(ECFCs-CM)on biological function of human dermal fibroblasts(HDFs). Methods Human cord blood derived-ECFCs were isolated and identified based on the previous studies. The cytokines in ECFCs-CM were detected using a cytokines antibody array. HDFs were cultured with ECFCs-CM,using serum free EBM-2 as control. The proliferation of HDFs was examined by Cell Counting Kit-8(CCK-8)and the migration was assessed by scratch test assay. The apoptosis of HDFs was detected by flow cytometry. Results The cells isolated from human cord blood demonstrated typical characteristics of ECFCs. The cytokines antibody array indicated that ECFCs-CM contained large amounts of secreted cytokines such as PDGF-BBand EGF. Compared with the control group,the HDFs cultured with ECFCs-CM showed improved proliferation and migration ability. The number of apoptotic cells was smaller than that of the control group under the environment of serum starvation. Conclusion ECFCs-CM can promote the proliferation and migration of HDFs and inhibit the apoptosis of HDFs under the environment of serum starvation.

Objective To investigate the clinical value of tumor's longest diameter and volume in assessing pathologic response in locally advanced breast cancer treated with neoadjuvant chemotherapy(NAC).Methods A prospective study was undertaken in women undergoing NAC for locally advanced breast cancer in order to determine the ability of quantitative MRI to assess the final pathologic response. 36 cases with pathologically confirmed locally advanced breast cancer who had been undergone four courses of NAC underwent preoperative breast MRI three times during the NAC. Pathologic response was assessed according Miller & Payne grading system, of which grade 4 and 5 defined as major histological response (MHR), and grade 1 to 3 as non-major histological response (NMHR). The tumor's longest diameters and volumes in MHR were compared with those in NMHR by Mann-Whitney U test before, after the second and fourth cycle of NAC. Concordance correlation coefficient (CCC) were assessed to evaluate the agreement between the two method. Receiver operating characteristic curve (ROC) analysis was carried out to assess the clinical value of tumor size and the change rate.Results Before,after the second and the fourth cycle of NAC, the difference of tumor's longest diameters and their first change rate between MHR [(2.75±1.16) cm,(2.19±1.07) cm ,(1.58±0.75) cm and (21.70±15.09)%]respectively,and NMHR [(2.71±1.10) cm,(2.33±0.90) cm,(2.01±0.94) cm and (11.68±10.27)%] respectively were not significant(P>0.05). The second change rate of tumor's longest diameter in MHR [(39.00±15.38)% ] was significantly higher than that in NMHR[ (25.83±21.77)% ](P=0.04). Before, after the second and the fourth cycly of NAC, the differences of tumor volumes and their first change rate between MHR [ median 14.00 cm~3 ( range 2.96～83.41 cm~3 ) , median 7.31 cm~3 (range 0.05～55.35 cm~3), median 2.69 cm~3 (range 0～33.40 cm~3 ) , median 48.65% (range 33.64%～98.48%) ] and NMHR [median 4.25 cm~3 (range 4.78～106.55 cm~3), median 10.53 cm~3 (range 1.72～42.85 cm~3), median 7.56 cm~3 (range 0.68～156.58 cm~3), median 52.04% (range-35.79%～78.31%) ] were not significant. The second changing rate of tumor's volume in MHR [median 85.37% (ranged 27.52%～100%)] was significantly higher than that in NMHR [median 68.80% (ranged -289.60%～94.24%)](P=0.01). CCC was computed before and (0.82), after the second cycle (0.67) and after the fourth cycle (0.55), in all examinations pooled together (0.78).The second change rates of tumor's longest diameter and volume were equal to predict the final pathologic response, and the area under curve were 0.75 and 0.80, respectively (P=0.61). Conclusion The agreement between the tumor's longest diameters and tumor's volumes is good in all in breast carcinoma. The assessment efficacy of the change rate of tumor's longest diameter and that of volume were low.

Objective To Compare different methods of quantitative breast density measurement.Methods The study included sixty patients who underwent both mammography and breast MRI. The breast density was computed automatically on digital mammograms with R2 workstation. Two experienced radiologists read the mammograms and assessed the breast density with Wolfe and ACR classification respectively. Fuzzy C-means clustering algorithm (FCM) was used to assess breast density on MRI. Each assessment method was repeated after 2 weeks. Spearman and Pearson correlations of inter- and intrareader and intermodality were computed for density estimates. Results Inter- and intrareader correlation of Wolfe classification were 0. 74 and 0. 65, and they were 0. 74 and 0. 82 for ACR classification respectively.Correlation between Wolfe and ACR classification was 0. 77. High interreader correlation of 0. 98 and intrareader correlation of 0. 96 was observed with MR FCM measurement. And the correlation between digital mammograms and MRI was high in the assessment of breast density (r = 0. 81, P ＜ 0. 01). Conclusion High correlation of breast density estimates on digital mammograms and MRI FCM suggested the former could be used as a simple and accurate method.