AIM: To study the effects of insulin and glucose on tissue-type plamingen activator (tPA) and its inhibitor-1 (PAI-1) secretion in cultured human endothelial cells. METHODS: Human endothelial cell line ECV-304 was cultured with glucose and/or insulin at different concentrations with or without hypoxic exposure. RESULTS: The tPA, PAI-1 secretion and ratio of tPA/PAI-1 increased in endothelial cells during hypoxia. Insulin and glucose increased the tPA and PAI-1 secretion in endothelial cells exposed to hypoxia, and increase in tPA/PAI-1 ratio was also observed at 4 h and 8 h. CONCLUSION: Hypoxia stimulates the release of tPA and PAI-1. Insulin and glucose also stimulate the tPA and PAI-1 secretion during hypoxia.

To assess the changes of sarcolemma Na+/K+ ATPase (CMNKA) and sarcoplasmic reticulum membrane Ca2+ ATPase (SERCA) activities after stem cells transplantation in heart failure. Rabbit was used as heart failure model by intravenously injecting adriamycin. Autologous bone marrow mononuclear cells (BMCs), bone marrow mesenchymal stem cells (MSCs) or skeletal myoblasts (SMs) were introduced into coronary arteies through the root of aorta when two balloons occluding just above sinus of Valsalva. After 4 weeks, left ventricular ejection fraction (LVEF)was evaluated by echocardiography, and the activities of CMNKA and SERCA were measured by colorimeter. In BMCs (n=8)and MSCs (n=8) group, LVEF were significantly improved (P < 0.05). No significant improvement were seen in SMs group (n=6) compared to sham group (n=8). The CMNKA activity in all stem cells groups was significantly increased compared to sham group (P < 0.05). Meanwhile, in comparison with sham group, the incremental tendencies of SERCA activity were seen in stem cells groups. In conclusion, stem cells transplantation could increase the activities of CMNKA and SERCA in heart failure, a possible mechanism to improve heart function.
Animals ; Doxorubicin ; Female ; Heart Failure ; chemically induced ; enzymology ; therapy ; Male ; Myocardium ; enzymology ; Rabbits ; Random Allocation ; Sarcolemma ; enzymology ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Stem Cell Transplantation

To investigate the feasibility of introcoronary cell infusion into nonischemic heart failure (HF) heart and whether different types of stem cell transplantation would affect heart function to a similar degree. Japanese white ears rabbits were used as HF models by intravenous injection adriamycin. Autologous bone marrow mononuclear cells(BMCs), bone marrow stromal cells (MSCs), skeletal myoblasts (SMs) or culture medium were infused into coronary arteries respectively by occluding the root of ascending aorta. The mortality during and 4 weeks after the procedure the mortality was 7.1% and 16.7% respectively. After 4 weeks, the ejection fraction (EF) in BMCs group had significant improvement (P < 0.05, n=8). No significant difference was seen in MSCs (n =8), SMs (n=6) and sham groups (n=8) compared with pretransplantation (P > 0.05). In sham group,the left ventricular endostolic diameter (LVED) had significant enlargement (P < 0.05), No significant difference was seen in MBCs, MSCs and SMs groups compared with pretransplantation (P > 0.05). Immunofluorescence revealed de novo expression of cardiac troponin I in BMCs and MSCs groups, cardiac troponin I was not detected in SMs group. In conclusions, intracoronary cell transplantation could provide effective cell delivery into dilated cardiomyopathy hearts and could be a useful strategy for treating CHF, BMCs cell transplantation may be the first choice in all the above cell types.
Animals ; Bone Marrow Transplantation ; Chronic Disease ; Coronary Vessels ; Heart Failure ; physiopathology ; surgery ; Infusions, Intra-Arterial ; Mesenchymal Stem Cell Transplantation ; Myoblasts, Skeletal ; transplantation ; Rabbits ; Random Allocation ; Stem Cell Transplantation ; methods ; Troponin ; metabolism ; Ventricular Function, Left ; physiology

Stem cells transplantation is a promising strategy for treating myocardial infarction and/or chronic heart failure; however, with respect to nonischemic heart failure, there are some limitations inherent in the current methods of transplantation. In this study, we investigated the feasibility of a novel method, i. e. transplantation through the root of aorta when the ascending aorta occluded above the sinus aortae. Japanese white ears rabbits were used as chronic heart failure models by intravenous injection of adriamycin. Autologous bone marrow mononuclear cells (MNC) were infused into the root of aorta when the ascending aorta was occluded by a couple of balloons above the sinus aortae. After 4 weeks, ejection fraction was significantly improved in MNC group. In conclusion, we have developed a unique method for efficient and safe cell transplantation based on infusion in aorta. This method, potentially suitable for nonischemic heart failure and could be used to achieve even and global supply of cells in heart.
Animals ; Aorta ; surgery ; Doxorubicin ; Feasibility Studies ; Heart Failure ; chemically induced ; surgery ; Rabbits ; Stem Cell Transplantation ; methods

Objective To investigate whether SIS3, a specific inhibitor of Smad3 phosphorylation, can reverse the stemness of multidrug?resistant ( MDR ) hepatocellular carcinoma cells. Methods MDR HCC Huh7. 5. 1/ADM cell lines were developed by exposing parental cells to stepwise increasing concentrations of ADM. CCK?8 assay was used to determine the cellular sensitivity of various anticancer drugs. Flow cytometry ( FCM) was used to analyze the expression level of cancer stem cell marker CD133. Clone formation assay and mouse subcutaneous xenograft tumors were used to investigate the tumorigenicity in vitro and in vivo. Western blotting ( WB) was used to analyze the changes of expressions of CD133, Smad3, Bcl?2, Bax and p?Smad3 in different conditions. Results ADM treatment of HCC cells in vitro resulted in a development of subline, Huh7. 5. 1/ADM cells, with CSC phenotypes: stable MDR phenotype ( besides ADMc Huh7.5.1/ADM cells were also more resistant to some other anticancer drugs including VCR, MMC and CTX ) (IC50:0.215±0.018 vs. 0.123± 0.004, 0.145±0.009 vs. 0.014±0.002, 1.021± 0.119 vs. 0.071± 0.006, 27.007±1.606 vs. 1.919±0.032)(unit: μg/ml)(P<0.05). Huh7.5.1/ADM cells enriched the cancer stem?like cell fraction (CD133?positive subpopulation) (76.06±2.948% vs. 25.38±4.349%)(P<0.05) , had stronger tumorigenicity in vivo and colony formation ability, and activated the Smad3 activity. Inhibition of Smad3 activity by SIS3 decreased stemness of the Huh7. 5. 1/ADM cells: CD133?positive subpopulation (48.49±2.304% vs. 76.06±2.948%)(P<0.05); ADM IC50: (0.112±0.019 vs. 0.215± 0.018), VCR IC50(0.065±0.013 vs. 0.145±0.009), MMC IC50(0.749±0.121 vs. 1.021±0.119), CTX IC50 (10.576±1.248 vs. 27.007±1.606)(unit:μg/ml)(P<0.05), and decreased tumorigenicity and colony formation ability. Conclusion SIS3 as a specific inhibitor of Smad3 signal is involved in the stemness of
multidrug resistant hepatocellular carcinoma cells.

Objective To investigate whether SIS3, a specific inhibitor of Smad3 phosphorylation, can reverse the stemness of multidrug?resistant ( MDR ) hepatocellular carcinoma cells. Methods MDR HCC Huh7. 5. 1/ADM cell lines were developed by exposing parental cells to stepwise increasing concentrations of ADM. CCK?8 assay was used to determine the cellular sensitivity of various anticancer drugs. Flow cytometry ( FCM) was used to analyze the expression level of cancer stem cell marker CD133. Clone formation assay and mouse subcutaneous xenograft tumors were used to investigate the tumorigenicity in vitro and in vivo. Western blotting ( WB) was used to analyze the changes of expressions of CD133, Smad3, Bcl?2, Bax and p?Smad3 in different conditions. Results ADM treatment of HCC cells in vitro resulted in a development of subline, Huh7. 5. 1/ADM cells, with CSC phenotypes: stable MDR phenotype ( besides ADMc Huh7.5.1/ADM cells were also more resistant to some other anticancer drugs including VCR, MMC and CTX ) (IC50:0.215±0.018 vs. 0.123± 0.004, 0.145±0.009 vs. 0.014±0.002, 1.021± 0.119 vs. 0.071± 0.006, 27.007±1.606 vs. 1.919±0.032)(unit: μg/ml)(P<0.05). Huh7.5.1/ADM cells enriched the cancer stem?like cell fraction (CD133?positive subpopulation) (76.06±2.948% vs. 25.38±4.349%)(P<0.05) , had stronger tumorigenicity in vivo and colony formation ability, and activated the Smad3 activity. Inhibition of Smad3 activity by SIS3 decreased stemness of the Huh7. 5. 1/ADM cells: CD133?positive subpopulation (48.49±2.304% vs. 76.06±2.948%)(P<0.05); ADM IC50: (0.112±0.019 vs. 0.215± 0.018), VCR IC50(0.065±0.013 vs. 0.145±0.009), MMC IC50(0.749±0.121 vs. 1.021±0.119), CTX IC50 (10.576±1.248 vs. 27.007±1.606)(unit:μg/ml)(P<0.05), and decreased tumorigenicity and colony formation ability. Conclusion SIS3 as a specific inhibitor of Smad3 signal is involved in the stemness of
multidrug resistant hepatocellular carcinoma cells.

Objective:To evaluate direct and indirect costs for schizophrenia,major depressive disorder(MDD)and bipolar disorder,and to compare their differences of cost composition,and to explore the drivers of the total costs.Methods:A total of 3 175 inpatients with schizophrenia,MDD,and bipolar disorder were recruited.In-patient's self-report total direct of medical costs outpatient and inpatient,out-of-pocket costs,and direct non-medical costs were regarded as direct costs.Productivity loss and other loss caused by damaging properties were defined as indirect costs.The perspectives of this study included individual and societal levels.Multivariate regression analysis was applied for detecting the factors influencing disease costs.Results:The total cost of schizophrenia was higher than those of MDD and bipolar disorder at individual and societal levels.The indirect costs of three mental disorders were higher than the direct costs,and the indirect cost ratio of bipolar disorder was higher than those of schizophre-nia and MDD.Age,gender,working condition and marital status(P＜0.05)were the important drivers of total costs.Conclusion:The economic burden of the three mental disorders is relatively heavy.Schizophrenia has heaviest disease burden,and the productivity loss due to mental disorders is the driving force of the soaring disease cost

Objective:To compare demographic characteristics,clinical characteristics,therapeutic characteris-tics and physiological indicators of patients with bipolar Ⅰ disorder and bipolar Ⅱ disorder.Methods:A total of 381 patients with bipolar disorder(BD)diagnosed by the Diagnostic and Statistical Manual of Mental Disorders 5 th Edi-tion(DSM-5)were selected,including 302 patients with BD-Ⅰ(79.27％),74 patients with BD-Ⅱ(19.42％)and 5 patients with other specific and related disorders(1.31％).Demographic and clinical characteristics were collected with self-designed clinical information questionnaire.Multivariate logistic regression and multivariate linear regres-sion analysis were used for analysis.Results:Compared with patients with BD-Ⅱ,patients with BD-Ⅰ had more risk to have psychotic features(OR=5.75,95％CI:2.82-11.76),longer disease duration,and more repeated transcra-nial magnetic therapy(OR=3.09,95％CI:1.02-9.35),higher uric acid,total cholesterol and high-density lipo-protein.BD-Ⅰ in Han nationality was more common(OR=11.50,95％CI:1.76-75.30),and had lower education level(OR=10.22,95％CI:1.16-89.77),and less family history of psychosis(OR=2.34,95％CI:1.01-5.42).Conclusion:There are significant differences between BD-Ⅰ and BD-Ⅱ in demographic and clinical charac-teristics,treatment status,and physiological indicators,which could provide clues for exploring the pathogenesis of bipolar disorder.

OBJECTIVETo investigate whether SIS3, a specific inhibitor of Smad3 phosphorylation, can reverse the stemness of multidrug-resistant(MDR) hepatocellular carcinoma cells.

METHODSMDR HCC Huh7.5.1/ADM cell lines were developed by exposing parental cells to stepwise increasing concentrations of ADM. CCK-8 assay was used to determine the cellular sensitivity of various anticancer drugs. Flow cytometry (FCM) was used to analyze the expression level of cancer stem cell marker CD133. Clone formation assay and mouse subcutaneous xenograft tumors were used to investigate the tumorigenicity in vitro and in vivo. Western blotting (WB) was used to analyze the changes of expressions of CD133, Smad3, Bcl-2, Bax and p-Smad3 in different conditions.

RESULTSADM treatment of HCC cells in vitro resulted in a development of subline, Huh7.5.1/ADM cells, with CSC phenotypes: stable MDR phenotype (besides ADMc Huh7.5.1/ADM cells were also more resistant to some other anticancer drugs including VCR, MMC and CTX ) (IC50: 0.215 ± 0.018 vs. 0.123 ± 0.004, 0.145 ± 0.009 vs. 0.014 ± 0.002, 1.021 ± 0.119 vs. 0.071 ± 0.006, 27.007 ± 1.606 vs. 1.919 ± 0.032) (unit: µg/ml) (P<0.05). Huh7.5.1/ADM cells enriched the cancer stem-like cell fraction (CD133-positive subpopulation) (76.06 ± 2.948% vs. 25.38 ± 4.349%) (P<0.05), had stronger tumorigenicity in vivo and colony formation ability, and activated the Smad3 activity. Inhibition of Smad3 activity by SIS3 decreased stemness of the Huh7.5.1/ADM cells: CD133-positive subpopulation (48.49 ± 2.304% vs. 76.06 ± 2.948%) (P<0.05); ADM IC50: (0.112 ± 0.019 vs. 0.215 ± 0.018), VCR IC50 (0.065 ± 0.013 vs. 0.145±0.009), MMC IC₅₀ (0.749 ± 0.121 vs. 1.021 ± 0.119), CTX IC50 (10.576 ± 1.248 vs. 27.007 ± 1.606) (unit: µg/ml) (P<0.05), and decreased tumorigenicity and colony formation ability.

CONCLUSIONSIS3 as a specific inhibitor of Smad3 signal is involved in the stemness of multidrug resistant hepatocellular carcinoma cells.

AC133 Antigen ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antigens, CD ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Glycoproteins ; metabolism ; Heterografts ; Humans ; Isoquinolines ; pharmacology ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; Mice ; Neoplasm Proteins ; metabolism ; Neoplastic Stem Cells ; drug effects ; Peptides ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyridines ; pharmacology ; Pyrroles ; pharmacology ; Smad3 Protein ; antagonists & inhibitors ; metabolism ; Tumor Stem Cell Assay ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE： To screen the quality control components of Chrysanthemum morifolium based multiple component metabolism， and study its network pharmacology effect. METHODS： The water extract of C. morifolium was prepared. A total of one rats were selected， water extract of C. morifolium was perfused in jejunum segment after abdominal anesthesia； plasma sample 1 was collected by double perfusion collection. Other 3 rats were given water extract of C. morifolium intragastrically， and plasma sample 2 was collected by abdominal aorta blood collection. UPLC-MS/MS was used to analyze water extract of C. morifolium and plasma sample component， and prototype blood-entry component in water extract of C. morifolium was identified after metabolism. TCMSP and Swiss Target Prediction database were used to screen the core target of prototype blood-entry component. DAVID database was used to enrich the related pathways of core target. The quality control components were screened according to topological parameters. Cytoscape software was used to analyze pharmacological effect of quality control components of C. morifolium. RESULTS： After UPLC-MS/MS analysis， 27 compounds were identified in water extract of C. morifolium， among which there were 12 prototype blood-entry components. After network pharmacology analysis， 7 quality control components were identified， i.e. cosmosiin， apigenin-7-O-glucuronide， luteolin， tilianin， apigenin， hesperetin， acacetin. It was possible to treat cancer， cardiovascular and cerebrovascular diseases， and neurological diseases by acting on metabolic pathway， cancer related pathway， signal transduction related pathway， adipocyte lipolysis regulatory pathway， etc. CONCLUSIONS： The study screen the possible quality control components of water extract of C. morifolium. The theoretical pharmacological effect of it can be clarified through network pharmacology， which can provide a new idea for the utilization of C. morifolium.