Objective To investigate the effects of rapamycin on the number and function of peripheral blood endothelial progenitor cells (EPCs). Methods Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7-day culture, adherent cells were treated with rapamycin in a series of final concentrations of 1.0, 2.0, 5.0?g/ml for 6, 12, 24, and 48h. EPCs were identified as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining as demonstrated under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of VEGFR-2, AC133 and CD34 with flow cytometry. EPCs proliferation and migration were assayed with MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating them on fibronectin-coated dishes, and then adherent cells were counted. In vitro vasculogenesis activity was assayed by in vitro vasculogenesis kit. Results Incubation of isolated human MNCs with rapamycin resulted in a decrease in the number of EPCs, and rapamycin also decreased EPCs proliferative, migratory, adhesive and in vitro vasculogenesis capacity in both concentration and time dependent manners. Conclusion Rapamycin decreases the number, proliferative, migratory, adhesive and in vitro vasculogenesis capacity of EPCs.

Objective To investigate the effect of high glucose on the number and proliferation, migration and adhesion of peripheral endothelial progenitor cells (EPCs). Methods Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin coated culture dishes. After 7 days of culture, several groups of attached cells were incubated with glucose in a series of concentrations (15, 25, 35, 45mmol/L) for different durations (6, 12, 24 and 48h). EPCs were characterized as adherent cells which were double positive for DiLDL uptake and lectin binding by direct fluorescent staining demonstrated under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of KDR, VEGFR 2 and AC133 with flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating MNCs on fibronectin coated dishes, and then the adherent cells were counted. Results Incubation of isolated human MNCs with high glucose concentration decreased the number of EPCs, and this effect was most prominant when glucose concentration was 45mmol/L, and incubated for 24 hours (approximately 1 fold decrease, P

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0.05).Compared with SHR group,left ventricular weight mass index decreased significantly in SHR-A group(P

Objective To investigate the real-time regulatory effects of IFN-γ, programed death ligand 2(PDL2) and janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway on the adherence, proliferation and migration of human placenta-derived mesenchymal stem cells(hPMSCs) based on a finding that IFN-γ could enhance the expression of PDL2 in hPMSCs through JAK/STAT signaling pathway.Methods hPMSCs were isolated by using enzyme digestion method and then co-cultured with IFN-γ, anti-PDL2 monoclonal antibody (anti-PDL2 McAb) and JAK inhibitor, respectively.Real-time cell analysis (RTCA) was used to detect the dynamic changes in the adherence, proliferation and migration of hPMSCs following various interventions.Results IFN-γ remarkably suppressed hPMSCs proliferation during the period from 40 hours to 80 hours after intervention and also inhibited the non-targeted migration of hPMSCs.However, hPMSCs adherence was not affected by IFN-γ.Co-culturing hPMSCs with anti-PDL2 McAb significantly enhanced hPMSCs adhesion and inhibited their non-targeted migration, but had no significant effect on hPMSCs proliferation.Furthermore, the proliferation of hPMSCs co-cultured with IFN-γ and anti-PDL2 McAb was significantly inhibited as compared with that of anti-PDL2McAb treatment group.The adhesion, migration and proliferation of hPMSCs were significantly inhibited after co-culturing them with JAK inhibitor.Conclusion IFN-γ can remarkably suppress the proliferation and migration of hPMSCs.PDL2 can enhance the migration and inhibit the adhesion of hPMSCs.JAK/STAT signaling pathway is involved in regulating the adhesion, migration and proliferation of hPMSCs.

Objective To investigate the correlations between the soluble form of B7-H3 ( sB7-H3) and the cytokines of IL-17 and IL-8 in serum samples from patients with primary hepatocellular carcino-ma ( HCC) and to evaluate their clinical values for early diagnosis of HCC.Methods Serum samples were collected from 63 patients with HCC and 50 healthy subjects.The expression of sB7-H3, IL-17 and IL-8 in serum samples were detected by ELISA.Receiver operating characteristic ( ROC) curve was generated to an-alyze the diagnostic values of sB7-H3, IL-17 and IL-8 for hepatoma.The logistic regression model was used to predict the probability of hepatoma by using sB7-H3, IL-17 and IL-8 in combination.Results The levels of sB7-H3, IL-17 and IL-8 in serum samples collected from the patients with HCC were significantly higher than those from healthy subjects.A positive correlation was found between the levels of sB7-H3 and IL-17 in serum samples from patients with HCC.No correlation was found between sB7-H3 and IL-8.A negative cor-relation was found between the levels of IL-17 and IL-8 in serum samples from patients with HCC.ROC curve analysis showed that the area under the curve (AUC) of sB7-H3, IL-17 and IL-8 were 0.832, 0.657 and 0.953, respectively, indicating the statistical significance of them for the diagnosis of HCC.The logistic regression showed that the AUC, diagnostic sensitivity and specificity of the regression model PRE in the pre-diction of HCC were 0.960, 91.30% and 94.29%, respectively, which was much better than using the three indicators alone.Conclusion The levels of sB7-H3 were positively correlated to the levels of IL-17 in serum samples from patient with HCC.The logistic regression model of combination of sB7-H3, IL-17 and IL-8 obtained in this study could be used for early clinical diagnosis of HCC in the future.

AIM: The aim of this study was to study the changes of rabbit heart electrophysiological properties caused by increasing left ventricular afterload, and to assess the effects of streptomycin or verapamil on these changes. METHODS: The rabbit heart preparation in situ was used,and the afterload of left ventricle was increased by clipping in part the root of ascending aorta. The changes of heart electrophysiological parameters including relative refractory period (RRP),effective refractory period (ERP),monophasic action potential duration (MAPD_ 90 ) and ventricular fibrillation threshold(VFT) were observed before and after altering the afterload of left ventricle and were compared in the absence and presence of streptomycin or verapamil. RESULTS: The rising of left ventricular afterload [(72?11)mmHg] led to shortening of RRP,ERP and MAPD_ 90 ,and to descent of VFT ( P 0.05) except increasing of VFT ( P

Objective To understand Shenzhen Longgang,guangming and longhua new district four district hospital ICU pa-tients with secondary pulmonary tuberculosis merger lower respiratory infection of pathogenic bacteria distribution and drug resistance status of provide a reference for clinical diagnosis and rational use of antibiotics therapy.Methods Random selec-tion from February 2013 to October 2015 in the three district hospital ICU diagnosis of secondary pulmonary tuberculosis patients with lower respiratory infection in 593 cases of sputum specimen pathogenic bacteria culture and drug susceptibility results were retrospectively analyzed.Results 593 cases of ICU secondary pulmonary tuberculosis patients with respiratory tract infection of the communist party of China isolated 617 strains of pathogenic bacteria,fungi accounted for 49.6% (306/617),gram negative bacilli accounted for 40.4% (249/617),gram positive cocci accounted for 10.0% (62/617).Fungal in-fection main pathogens for white smooth candida yeast and candida yeast,respectively accounted for 44.2% (273/617)and
4.5% (28/617),gram negative bacillus mainlyKlebsiellaPneumoniae,Pseudomonasaeruginosa,and H.influenzae,respec-tively accounted for 16.7% (103/617),12.0% (74/617)and 7.3% (45/617),gram-positive cocci mainly for Saphylococcus aureus and Epidermisstaphylococcus and Hemolyticstaphylococci,respectively accounted for 4.5% (28/617),3.2% (20/617)and 0.9% (5/617).Pathogenicbacteria isolated from the multiple drug resistant bacteria,present different levels of resistance to commonly used antimicrobial agents.Conclusion ICU patients with secondary pulmonary tuberculosis merger of lower respiratory tract infection pathogens to fungi and gram-negative bacilli,the most commonWhite candida,Klebsiella pneumoniae,and Pseudomonasaeruginosa,and different levels of resistance to commonly used antimicrobial agents.

AIM: To investigate whether homocysteine (Hcy) has influences on endothelial progenitor cell (EPCs) number and activity from peripheral blood. METHODS: Total mononuclear cells (MNCs) were plated on fibronectin-coated culture dishes and cultured for 7 days, and then attached cells were stimulated with Hcy or vehicle control for 6 h, 12 h, 24 h and 48 h. The adhesion, proliferation, migration and in vitro vasculogenesis activity of EPCs were assayed, respectively. RESULTS: Incubation of isolated human MNCs with Hcy dose and time-dependently decreased the number of EPCs with maximum at 200 (?mol/L) for 24 hours (35.7?6.7 vs 62.5?10.6, P

Objective:To investigate the effect of ubiquitin binding enzyme 2T (UBE2T) on the radiosensitivity of lung adenocarcinoma and unravel its possible mechanism.Methods:A total of 45 patients pathologically diagnosed with different stages of lung adenocarcinoma and treated with radiotherapy in the Second Affiliated Hospital of Zunyi Medical University from March, 2019 to December, 2021 were enrolled, and the efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST1.1). All patients were divided into radiosensitive group ( n=25) and radioresistant group ( n=20). Radiosensitive group was complete remission (CR)+partial remission (PR), and radioresistant group was stable disease (SD) + progression disease (PD). Immunohistochemistry (IHC) was used to calculate the score based on the staining intensity and the number of positive cells. Chi-square test was combined to analyze the correlation between the expression level of UBE2T in paraffin specimens of lung adenocarcinoma patients and the radiosensitivity of patients. Lentivirus UBE2T-interfered (UBE2Tsh) A549 and UBE2T-overexpressed SPC-A-1 lung adenocarcinoma cells and their respective controls were constructed for irradiation and colony formation assay. The survivor fraction curve was fitted by single-hit multi-target model. The DNA double-strand break (DSB) marker γH2AX foci were detected by immunofluorescence (IF). The expression levels of UBE2T, γH 2AX and Rad51 proteins were detected by Western blot. Cell cycle and apoptosis rate of A549 were determined by flow cytometry. Binary variables were statistically analyzed by Fisher's exact probability method and measurement data were assessed by t-test. Results:High-expression level of UBE2T was correlated with the radiosensitivity of lung adenocarcinoma patients ( P<0.05). UBE2Tsh improved the radiosensitivity of A549 lung adenocarcinoma cells, and the sensitizing enhancement ratio (SER) was 1.795. UBE2T overexpression decreased the radiosensitivity of SPC-A-1 lung adenocarcinoma cells with an SER of 0.293. γH2AX foci number per cell were significantly increased in UBE2Tsh A549 cells after irradiation ( P<0.01) . Compared with the control group, the expression level of γH2AX protein was up-regulated ( P<0.01)and that of Rad51 protein was down-regulated in UBE2Tsh A549 cells after radiation ( P<0.001). Compared with the control group, the expression level of γH2AX protein was down-regulated ( P<0.05) and that of Rad51 protein was up-regulated in UBE2T overexpressed SPC-A-1 cells ( P<0.001). The proportion of UBE2Tsh A549 cells in G 2 phase was decreased ( P<0.01) and cell apoptosis was increased ( P<0.001). Conclusions:UBE2T might promote the radioresistance of lung adenocarcinoma cells by enhancing DNA DSB repair induced by radiotherapy, inducing cell cycle G 2 phase arrest, and reducing cell apoptosis.