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OBJECTIVE To study the effects of the curcumin derivative bisdemethoxycurcumin (BC) promoting neuronal differentiation of neuroblastoma cells Neuro-2a (N2a) in mice and its mechanism. METHODS The effects of BC (1, 2, 4, 6, 8, 10 μmol/L) on the viability of N2a cells were detected by MTT assay to determine the concentration range of drug treatment. The control group, retinoic acid (RA) group (10 μmol/L) and BC groups (1, 2 and 4 μmol/L) were set up, and the length of neural protrusions of the differentiated cells was measured and the cell differentiation rate was calculated after 48 h and 72 h of culture. Compared with 0 min group, Western blot was used to detect the phosphorylation levels of protein kinase B (Akt), extracellular- signal regulated kinase 1/2 (ERK1/2), and p38 mitogen-activated protein kinase (p38) proteins in cells treated by 4 μmol/L BC for 5, 15, 30, 60, 120 min. After intervention with inhibitors LY294002 (LY) and PD98059 (PD), the effects of BC on Akt and ERK1/2 protein phosphorylation levels and promoting neural differentiation were further validated. RESULTS According to the MTT experiment, the BC concentrations for subsequent induction of cell differentiation were determined to be 1, 2, and 4 μmol/L. After 48 hours of differentiation, compared with the control group, the cell differentiation rate in RA group and BC 1, 2 and 4 μmol/L groups, the length of cellular neural processes wjxhhxx413@163.com in the BC 4 μmol/L group significantly increased (P<0.05 or P<0.01);after inducing differentiation of BC for 72 hours,compared with the control group, the cell differentiation rate and the length of cellular neural processes in the RA group, the cell differentiation rate in the BC 4 μmol/L group, and the length of cellular neural processes in the BC 2 μmol/L group all significantly increased (P<0.05 or P<0.01).Compared with the 0 min group, the phosphorylation levels of Akt, ERK1/2, and p38 proteins in cells of the 5, 15, 30, 60 and 120 min groups increased to varying degrees after treated by 4 μmol/L BC, and some differences were statistically significant (P<0.05 or P<0.01). After adding the inhibitor LY/PD, compared with the BC group, the phosphorylation level of ERK1/2 protein in the PD+BC group cells were significantly reduced (P<0.01), and the cell differentiation rates in the LY group, LY+BC group, PD group, and PD+BC group was significantly reduced (P<0.01). CONCLUSIONS BC promotes N2a cell differentiation mainly by increasing cell differentiation rate and neural protrusion length. The mechanism may be related to the activation of mitogen-activated protein kinase/ ERK and PI3K/Akt signaling pathways.
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The infiltration of immune cells into the hepatocellular carcinoma microenvironment is the main reason why hepatocellular carcinoma patients are prone to carcinoma recurrence and the disease are incurable. Notably, the infiltration of Treg cells is the main trigger. Dahuang Zhechong pill (DHZCP) is a traditional Chinese herbal compound successful in the treatment of hepatitis and hepatocellular carcinoma. DHZCP can heal and nourish while slowing the onset of the disease, thereby strengthening the body's immune function. It can localize tumors and ultimately achieve the goal of eliminating tumors. In this study, an orthotopic liver cancer model of mice was used to explore the mechanism of DHZCP enhancing anti-tumor immunity, which showed more Th1 cells in the peripheral blood and spleen after DHZCP treatment, while more IFN-γ was secreted to activate CD8+ T cells and Treg cell production was inhibited, thereby suppressing the growth of HCC. Finally, we also analyzed the potential components of DHZCP from the perspective of modern targets using network pharmacology methods and experimental results.
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Animales , Humanos , Ratones , Linfocitos T CD8-positivos , Carcinoma Hepatocelular/tratamiento farmacológico , Medicamentos Herbarios Chinos , Neoplasias Hepáticas/tratamiento farmacológico , Linfocitos T Reguladores , Microambiente TumoralRESUMEN
Objective: To investigate the clinical characteristics and risk factors of postoperative atrial fibrillation (POAF) in patients with critical burns. Methods: A retrospective case series study was conducted. From January 2017 to December 2021, two hundred and twenty-seven critically burned aldult patients who met the inclusion criteria were admitted to Guangzhou Red Cross Hospital of Jinan University, including 173 males and 54 females, aged 19-83 (43±14) years. The admission years of patients were collected, and the percentage of patients complicated with POAF in each year was calculated. According to whether the patients were complicated with POAF or not, they were divided into POAF group (n=17) and non-POAF group (n=210). Following data were collected in patients in POAF group, including operation methods, duration of operation, intraoperative blood loss before occurrence of POAF each time, occurrence time and times of POAF, postoperative body temperature, blood pressure, hemoglobin, blood glucose, blood lactate, sepsis, and electrolyte, and type, duration, and treatment of POAF. General data of patients in the two groups including age, gender, burn reason, total burn area, full-thickness burn area, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and sepsis-related organ failure evaluation (SOFA) scores on admission, combined with underlying diseases (hypertension, diabetes, and other types of arrhythmias), and sepsis were collected and analyzed. The mortality and factors influencing the prognosis of patients in the two groups such as mechanical ventilation time, operations times, and burn intensive care unit (BICU) length of stay were also collected and analyzed. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, chi-square test or Kruskal-Wallis H test. The multivariate logistic regression analysis was performed on the general data with statistically significant differences between the two groups, and the independent risk factors influencing the onset of POAF in 227 patients with critical burns were screened. Results: From 2017 to 2021, the percentage of critically burned patients complicated with POAF increased year by year. In POAF group, eschar debridement in limbs was the main surgical procedure prior to POAF complication, with the operation time of (3.5±1.2) h and the intraoperative blood loss volume of (365±148) mL.The POAF occurred 25 times in total in patients of POAF group, mostly within one week after the injury and within 6 hours after the operation with most of these patients having POAF only once. When POAF happened, the patients were often complicated with hypothermia, anemia, hyperglycemia, high blood lactate, sepsis, and electrolyte disturbance, and few patients had complications of hypotension. The POAF lasted (5±3) h, with all being paroxysmal atrial fibrillation, and most of POAF patients were reverted to sinus rhythm after amiodarone intervention. Most patients in the two groups suffered from flame burn, and the gender, age, and SOFA score on admission of patients in the two groups were similar (P>0.05); the APACHEⅡ score on admission, total burn area, full-thickness burn area, incidence proportion of sepsis, combined with diabetes and hypertension and other types of arrhythmias of patients in POAF group were significantly higher or larger than those in non-POAF group (t=3.47, with χ2 values of 7.44, 10.86, 12.63, 14.65, 6.49, and 7.52, respectively, P<0.05 or P<0.01). The full-thickness burn area, combined with other types of arrhythmias, and sepsis were the independent risk factors for POAF in 227 critically burned patients (with odds ratios of 4.45, 0.04, and 3.06, respectively, with 95% confidence intervals of 2.23-8.87, 0.01-0.22, and 1.77-5.30, respectively, P<0.01). Compared with those in non-POAF group, the mechanical ventilation time, BICU length of stay, number of operations, and mortality rate of patients in POAF group were significantly increased (Z=3.89, Z=2.57, t=3.41, χ2=3.72, P<0.05 or P<0.01). Conclusions: POAF is a common postoperative complication in critically burned patients, and the incidence is increasing year by year, which seriously affects the prognosis of patients. The full-thickness burn area together with other types of arrhythmias and sepsis are the high-risk factors for POAF complication in patients with critical burns.
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Femenino , Humanos , Masculino , Fibrilación Atrial/etiología , Pérdida de Sangre Quirúrgica , Hipertensión , Lactatos , Pronóstico , Estudios Retrospectivos , Factores de Riesgo , SepsisRESUMEN
There are many chemical components in the volatile oil of Dictamni Cortex. The complex network relationship of "component-target-disease" can be revealed by using the network pharmacology method, and the mechanism of the efficacy of Dictamni Cortex can be revealed. In this study, we used Swiss Target Prediction database to predict the target of action, STRING database to build protein interaction network, and Cytoscape software to build "component-target-disease" network. The results showed that the antibacterial, anti-inflammatory and antiallergic effects of Dictamni Cortex were closely related to the components of thymol methyl ether, elemenol, anethole, and the related targets of each component were cross-linked to play a multi-target pharmacodynamic role. This study laid a foundation for the study of the effective substance basis and quality control evaluation of the Dictamni Cortex, and provided a scientific basis for further revealing its mechanism.
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Dictamnus/química , Medicamentos Herbarios Chinos/farmacología , Aceites Volátiles/farmacología , Mapas de Interacción de Proteínas , Control de Calidad , Programas InformáticosRESUMEN
Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.
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Animales , Ratones , Ácido Acético , Cromatografía Líquida de Alta Presión , Curcuma , Química , Medicamentos Herbarios Chinos , Química , Farmacología , Espectrometría de Masas , Rizoma , Química , Trombosis , QuimioterapiaRESUMEN
Objective To understand the health literacy of diabetes prevention and control among community residents in Chongqing City, and to provide reference for development of health education and health promotion. Methods In July 2015, a random sampling method was used to select samples of residents aged 18 years and above in Chongqing. The health literacy questionnaire of urban and rural residents was used to investigate. The logistic regression model was used to analyze the factors affecting the health literacy of diabetes. Results The effective 892 respondents were collected from 1000 residents, and the effective rate was 89.20%. There were 440 men and 452 women; the average age was 61.71±12.29; there were 176 residents having diabetes prevention and health literacy, accounting for 19.73%. Multivariate logistic regression analysis showed that female (OR=1.679, 95%CI: 1.028-2.743), culture degree (ORjunior =3.891, 95%CI: 1.658-9.173; ORsecondary and high scho l =2.959, 95%CI:1.374-6.369; ORcol ege and above=5.319, 95% CI: 2.433-11.628) and the self assessment of health (ORgeneral=2.892, 95% CI:1.338-6.251) were higher in the diabetes prevention health literacy. The health education methods include medical staff on-site explanation, community lectures, watching video and propaganda columns were more popular. Conclusion The level of health literacy of diabetes was generally low in Chongqing. The community should focus on diabetes health education activities for men and low educated population. The community should carry out more activities about diabetes for residents, which helps to reduce or delay the occurrence of diabetes in the population.
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Objective: To investigate the effect of brain derived neurotrophic factor (BDNF) on mesenchymal stem cells (MSC) inhibiting follicular helper T cells (Tfh cells). Methods: The contents of indoleamine 2,3-dioxygenase (IDO), IL-10, TGF-β and IL-21 in MSC culture supernatant were detected by ELISA; The peripheral blood of healthy volunteers were collected, and lymphocyte in peripheral blood was separated by human lymphocyte separation solution; Co-cultures of MSC and lymphocyte were performed by Transwell chamber, and the proportion of CD4(+)CXCR5(+) Tfh cells and their subtypes were detected by flow cytometry. Results: ①The concentrations of IL-10, TGF-β, and IDO in the supernatant of BDNF group (BDNF-stimulated MSC) were higher than those of the control ones (adding PBS with the same volume) [IL-10: (42.1±4.4) ng/ml vs (19.3±2.1) ng/ml, t=4.761, P=0.009; TGF-β: (13.9±1.7) ng/ml vs (5.3±0.6) ng/ml, t=5.129, P=0.008; IDO: (441.3±56.9) ng/ml vs (226.7±37.6) ng/ml, t=3.130, P=0.035]; ②The comparisons between BDNF (co-culture of lymphocyte and BDNF-stimulated MSC) and MSC groups (co-culture of lymphocyte and MSC) were detailed as of follows: the proportion of CD4(+) CXCR5(+)Tfh cells were lower [(3.37±0.21)% vs (6.51±0.27)%, t=9.353, P<0.001], the proportion of CD4(+) CXCR5(+)CXCR3(+) CCR6(-) Tfh cells were higher [(41.14±2.04)% vs (26.72±2.57)%, t=4.383, P=0.012], CD4(+)CXCR5(+)CXCR3(-)CCR6(-) Tfh2 cells and CD4(+)CXCR5(+)CXCR3(-)CCR6(+) Tfh17 cells were lower [Tfh2: (30.16±5.38)% vs (43.26±4.11)%, t=4.426, P=0.012; Tfh17: (15.61±1.52)% vs (22.32±0.72)%, t=4.202, P=0.014], the proportion of CD4(+)CXCR5(+)Foxp3(+) Tfr cells were higher [(4.95±0.22)% vs (2.32±0.16)%, t=10.241, P<0.001], the concentration of IL-21 in the lymphocyte supernatant was lower [(0.28±0.03) ng/ml vs (0.85±0.08) ng/ml, t=6.675, P=0.003]. Conclusion: BDNF could enhance the inhibitory effect of MSC on Tfh cells through inhibiting the increasing of Tfh cells and the differentiations of Tfh2 and Tfh17 cells.
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Humanos , Factor Neurotrófico Derivado del Encéfalo , Diferenciación Celular , Citometría de Flujo , Células Madre Mesenquimatosas , Linfocitos T Colaboradores-InductoresRESUMEN
Notoginsenoside R1(NGR1),a critical compound in traditional herb Panax notoginseng, is a kind of estrogen receptor agonist.It is reported to exhibit anti-apoptotic,anti-oxidative and anti-inflammatory properties activity, so it is widely used for treatment of various diseases.In order to investigate the potential neuroprotective effect of NGR1 in hypoxic-ischemic brain damage(HIBD), primary cortical neurons were used in this study to establish oxygen-glucose deprivation/reoxygenation(OGD/R) injury models. They were treated with NGR1 and estrogen receptor inhibitor ICI-182780 respectively, then the neuronal survival, cell membrane integrity and apoptosis were assessed by MTT assay,lactate dehydrogenase test(LDH) and Hoechst 33342 stain respectively, while the protein expression levels of ATF6α,p-Akt,Akt,Bax and Cleaved Caspase-3 were measured by Western blotting. Results indicated that as compared with the blank control group,OGD/R could induce cell injury and apoptosis(P<0.05), reduce relative integrity of cell membrane(P<0.05), decrease protein expression of ATF6α,p-Akt(P<0.05), and increase protein expression of Bax and Cleaved Caspase-3(P<0.05) in the primary cortical cells. After NGR1 treatment, the expression levels of ATF6α,p-Akt were obviously increased, and the expression levels of Bax and Cleaved Caspase-3 and the apoptosis of neuron were decreased(P<0.05). However, these neuroprotective properties of NGR1 against ODG/R-induced cell damage could be blocked by ICI-182780. This finding indicated that NGR1 may protect the primary cortical neurons against OGD/R induced injury,and the mechanism may be associated with accelerating the activation of the ATF6/Akt signaling pathway via estrogen receptors.
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<p><b>OBJECTIVE</b>This study aimed to investigate the susceptibility of mice with streptozotocin(STZ)-induced diabetes mellitus (TIDM) to the uptake of pentavalent inorganic arsenic (iAsV) and the possible molecular mechanism.</p><p><b>METHODS</b>TIDM was induced in mice by STZ. TIDM and normal mice were treated with 15.0 mg/kg Na2HAsO4·12H2O by intragastric administration. Then, the concentrations of arsenic in various tissues were measured by atomic fluorescence spectrometry. The gene expression levels of Pit1 and Pit2 were quantified by real-time RT-PCR, and their protein levels were detected by Western blotting in mouse heart, kidney, and liver tissues.</p><p><b>RESULTS</b>The concentrations of arsenic in STZ-induced TIDM mouse tissues were higher at 2 h after intragastric administration of Na2HAsO4·12H2O. Compared with the levels in normal mice, PIT1 and PIT2, which play a role in the uptake of iAsV, were upregulated in the livers and hearts of TIDM mice. PIT1 but not PIT2 was higher in TIDM mouse kidneys. The upregulation of Pit1 and Pit2 expression could be reversed by insulin treatment.</p><p><b>CONCLUSION</b>The increased uptake of iAsV in TIDM mouse tissues may be associated with increased PIT1 and/or PIT2 expression.</p>
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Animales , Masculino , Ratones , Arsénico , Farmacocinética , Diabetes Mellitus Experimental , Metabolismo , Contaminantes Ambientales , Farmacocinética , Regulación de la Expresión Génica , Fisiología , Ratones Endogámicos ICR , Proteínas Cotransportadoras de Sodio-Fosfato de Tipo III , Genética , Metabolismo , Factor de Transcripción Pit-1 , Genética , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To study the effects of graphene quantum dots (GQDs) on hematopoietic system in rats.</p><p><b>METHODS</b>Thirty male SD rats were randomly divided into three groups (n = 10): control group, high dose group (10 mg/kg · d), low dose group (5 mg/kg · d), The rats in experimental group were intravenous injected with GQDs for 28 days and those in control group were injected with normal saline at the same volume. Routine blood and the function of liver and kidney were detected by instrument analysis. The cycle and apoptosis of bone marrow mononuclear cells (BMCs) were detected by FCM. The other three only healthy male SD rat bone marrow mononuclear cells (BMCs) were cultured by joining GQDs for 24 h, 48 h,72 h in vitro, the proliferation was assayed by CCK-8, the content of granulocyte macrophage colony stimulating factor (GM-CSF) from cultural supernatants were detected by ELISA.</p><p><b>RESULTS</b>The amount of red blood cell and concentration of hemoglobin from experimental group were increased significantly compared with those of control groups (P < 0.05), the concentration of triglyceride and high density lipoprotein were decreased. DNA synthesis period was prolonged (P < 0.01), there was no significant difference in apoptosis. BMCs were promoted proliferation clearly after using GQDs for 72 h (P < 0.05). The content of GM-CSF was increased (P < 0.01) .</p><p><b>CONCLUSION</b>GQDs may promote hematopoietic function in rats.</p>
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Animales , Masculino , Ratas , Apoptosis , Células de la Médula Ósea , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Metabolismo , Grafito , Farmacología , Hematopoyesis , Puntos Cuánticos , Química , Ratas Sprague-DawleyRESUMEN
<p><b>OBJECTIVE</b>To screen out main molecular target promoting human neural stem cells (NSCs) of ginsenoside Rg1 by using the gene chip technology.</p><p><b>METHOD</b>First, MTT assay was adopted to screen out the optimal concentration of Rg1-promoted NSC proliferation (120 mg x L(-1)). Then, on the 7th day after the Rg1-promoted NSC proliferation, the expression of target genes was observed by the gene chip technology. The most important target gene and signal transduction pathways were screened out through the data calculations.</p><p><b>RESULT</b>On the 7th day after the Rg1-promoted NSC proliferation, obtained 440 differential genes, 266 significantly upregulated genes and 174 significantly down-regulated genes. HES1 gene, CAMP (cyclic adenosine monophosphate)-PKA (protein kinase A) and PI3K (phosphatidylinositol 3 kinase)-AKT signal transduction pathways were closely related to the NSC proliferation.</p><p><b>CONCLUSION</b>The differentially expressed genes screened out by gene chip may provide new clues for studies on molecular mechanism of ginsenoside Rg1-promoted NSCs proliferation.</p>
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Humanos , Proliferación Celular , Ginsenósidos , Farmacología , Células-Madre Neurales , Biología Celular , Metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To observe the role of tumor necrosis factor α (TNF-α) in endothelial-mesenchymal transition (EnMT), and to explore the mechanism of fibrosis disease.</p><p><b>METHODS</b>Human umbilical vein endothelial cells (HUVEC) from umbilical cord of healthy fetus were isolated by enzymatic digestion and identified by immunofluorescence assay. The third to fifth generations of cultured HUVEC in logarithmic phase were harvested and seeded in 12-well plates and 6-well plates, and they were divided into control group (ordinary culture without any stimulation), 5, 10, 25, 50, and 100 ng/mL TNF-α groups (5, 10, 25, 50, 100 ng/mL of TNF-α was respectively added into the nutrient solution) according to the random number table, with three samples in each group. After being cultured for 72 hours, the cell morphology was observed under inverted phase-contrast microscope; the expression levels of coagulation factor VIII and α smooth muscle actin (α-SMA) were detected by immunofluorescence assay, and the ratios of numbers (absorbance values) of cells with expression of both factors were calculated. The mRNA expression levels of cadherin, α-SMA, and type I collagen were detected by RT-PCR (denoted as gray value ratio). Data were processed with one-way analysis of variance and LSD test.</p><p><b>RESULTS</b>(1) The shape of primary HUVEC was round, short-spindle, or flat, and cells grew vigorously in cobblestone appearance after passages. After being subcultured for 1, 2, 3, 4, 5 passage (s), the positive rate of coagulation factor VIII of HUVEC was respectively (85.5 ± 1.8)%, (88.1 ± 5.0)%, (93.6 ± 3.7)%, (92.9 ± 4.8)%, (89.5 ± 1.1)%, and they were significantly higher than that of primary HUVEC [(81.4 ± 3.8)%, with F values all equal to 7.481, P values all below 0.05]. (2) As compared with that in control group, the appearance of cells in 5, 10, 25, 50, and 100 ng/mL TNF-α groups was gradually transformed from round, short-spindle, or flat shape to long-spindle shape with reduced intercellular junction and larger intercellular gap along with the increase in the concentration of TNF-α. (3) The ratios of numbers and the absorbance values of coagulation factor VIII and α-SMA double positive cells in control group (0.055 ± 0.015, 0.078 ± 0.017) were significantly lower than those in 5, 10, 25, 50, and 100 ng/mL TNF-α groups (0.257 ± 0.106, 0.280 ± 0.129, 0.505 ± 0.059, 0.817 ± 0.035, 0.929 ± 0.101 and 0.437 ± 0.040, 0.456 ± 0.097, 0.496 ± 0.082, 0.787 ± 0.131, 0.885 ± 0.087, with F value respectively 45.009, 50.099, P values all below 0.01). (4) The expression levels of cadherin mRNA in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.70 ± 0.05, 0.63 ± 0.06, 0.60 ± 0.10, 0.45 ± 0.16, and 0.26 ± 0.14, and it was significantly lower in the latter four groups than in control group (0.83 ± 0.03, with F values all equal to 11.593, P < 0.05 or P < 0.01). The mRNA expression levels of α-SMA and collagen I in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.45 ± 0.10, 0.51 ± 0.16, 0.49 ± 0.12, 0.60 ± 0.09, 0.76 ± 0.03 and 0.38 ± 0.18, 0.45 ± 0.15, 0.52 ± 0.12, 0.66 ± 0.17, 0.76 ± 0.20, and they were significantly higher in the latter three groups than in control group (0.37 ± 0.14, 0.31 ± 0.12, with F value respectively 7.839, 2.898, P < 0.05 or P < 0.01).</p><p><b>CONCLUSIONS</b>TNF-α can obviously promote EnMT in a dose-dependent manner. EnMT may be another significant source of myofibroblasts that contributes to fibrotic tissue in scar formation.</p>
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Humanos , Diferenciación Celular , Células Cultivadas , Transición Epitelial-Mesenquimal , Células Endoteliales de la Vena Umbilical Humana , Biología Celular , Células del Estroma , Biología Celular , Factor de Necrosis Tumoral alfa , FarmacologíaRESUMEN
<p><b>OBJECTIVE</b>To observe the effects of ginsenoside Rg1 on the functional expression of human neural stem cells (hNSCs).</p><p><b>METHOD</b>The membrane electrophysiological properties and sodium and potassium ion channels in the hNSCs induced by Rg1 were analyzed using the whole-cell patch-clamp.</p><p><b>RESULT</b>On the 7th day, the neuron-like cells derived from ginsenoside Rg1 (20 mg x L(-1))-induced NSCs show: (1) The resting membrane potential: (-45.70 +/- 2.63) mV, the membrane capacitance: (26.89 +/- 1.91) pF, the membrane input impedance: (877.51 +/- 20.44) MH (P < 0.05 compared with the control group, respectively); (2) The detection rate of inward sodium current which is rapidly activated and inactivated in voltage-dependence was 50%, and its average peak value was (711.48 +/- 158.03) pA (P < 0.05 compared with the control group); (3) The outward potassium currents were composed of rapidly activated and inactivated transient outward potassium current and delayed rectifier outward potassium current, and its average peak value was (1 070.42 +/- 177.18) pA (P < 0.05 compared with the control group).</p><p><b>CONCLUSION</b>Ginsenoside Rg1 can promote the functional expression and maturity of hNSCs.</p>
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Humanos , Células Cultivadas , Expresión Génica , Ginsenósidos , Farmacología , Potenciales de la Membrana , Células-Madre Neurales , Biología Celular , Técnicas de Placa-Clamp , Extractos Vegetales , Farmacología , Canales de Potasio , Genética , Metabolismo , Canales de Sodio , Genética , MetabolismoRESUMEN
Objective To identify the candidate genes in the vicinity of a susceptibility locus (urinary albumin 1,UA-1) contributing to the development of albuminuria in type 2 diabetic KK/Ta mice. Methods Total RNA was extracted from the kidneys of KK/Ta (n=3) and BALB/c (n=2) mice at 20 weeks of age.The gene expression profile in kidney was investigated using the Affymetrix Murine Genome U74Av2 array.Competitive RT-PCR was used to confirm the differential expression of syndecan-4 which located in the vicinity of UA-1.Genome DNA was extracted from KK/Ta and BALB/c mice.DNA sequence analysis of the coding and promotor region of syndecan-4 gene was conducted. Results In the vicinity of the susceptibility locus (UA-1)contributing to the development of albuminuria in type 2 diabetic KK/Ta mice,10 candidate genes that showed differential expression were identified.Among them,the gene expression of syndecan-4in KK/Ta kidneys at 20 weeks of age was up-regulated by 26.1 times of age-matched BALB/c kidneys.Sequence analysis revealed two synonymous polymorphisms in the coding region (A93C and T216C) and three polymorphisms in the promoter region (-T263C,-T396C and -G669A) of the syndecan-4 gene.The TATA box was found at 321 bp upstream from the transcription start site,and the T263C polymorphism was located in the binding site of transcription factor Clox.Conclusions Syndecan-4 gene is mapped in the vicinity of the susceptibility locus contributing to the development of albuminuria in type 2 diabetes.The gene expression of syndecan-4 in KK/Ta kidneys is up-regulated than that in age-matched BALB/c kidneys at 20 weeks of age.Thus syndecan-4 may be one of the potential candidate genes responsible for diabetic nephropathy.Sequence differences in the promoter region influence the expression levels of syndecan-4 genes in KK/Ta kidneys.
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<p><b>AIM</b>To observe the effect for the model of PD and the transplantation of NSCs after injection of TSPG into mouse in advance.</p><p><b>METHODS</b>Firstly, we divided the mouse into 5 groups. For the group of 1-4, we established the model of PD with MPTP. For the group of 5, before the establishment of the model, we injected TSPG into mouse in advance for prevention. And then, we evaluated the effect by paralysis agitans score standard and praxiology marker. Secondly, we obtained the NSCs from the 7-12 week embryo cerebral cortex. Then we transplanted NSCs which pretreated by TSPG into the striatum of the 5 groups. After 60 days, we obtained the brain section, and detected the TH by ICC to analyse the differentiation status of NSCs.</p><p><b>RESULTS</b>The prevention of TSPG could decrease the neural cells damage by MPTP, and could protect the nervous system. After we transplanted NSCs into the striatum of Parkinson' s disease mouse, we found that for the group of 5, the paralysis agitans, auto-activity and memory function had the most distinct amelioration. And the number of dopaminergic neurons increased most transparently in brain section, and the neurons contact was the most enriched with the adjacent nervous cells.</p><p><b>CONCLUSION</b>TSPG can decrease the neural cells damage and can produce a marked effect in treatment of PD by transplanting NSCs invivo.</p>
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Animales , Humanos , Masculino , Ratones , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina , Terapia Combinada , Ratones Endogámicos C57BL , Células-Madre Neurales , Trasplante , Panax , Química , Enfermedad de Parkinson Secundaria , Terapéutica , Distribución Aleatoria , Saponinas , Usos TerapéuticosRESUMEN
The expression of tumor suppressor gene p16INK4a in mouse endometrium during early pregnancy and its possible role in blastocyst implantation were investigated in the present study. Real-time fluorescent quantitative PCR (FQ-PCR) and immunohistochemistry were applied to detect p16INK4a mRNA and protein expressions in endometrium of un-pregnant and pregnant mice on day 2, 3, 4, 5, 7, respectively. In addition, p16INK4a antibody was injected into the horns of uteri in pregnant mice on day 3 and its effect during blastocyst implantation was detected in vivo. The higher expressions of p16INK4a mRNA and protein were observed in pregnant mice compared with that in un-pregnant mice, with a steady increase from day 2 to day 5 and reaching the maximal level on day 5 of pregnancy and then decreasing. p16INK4a antibody decreased the number of implanted blastocysts compared with that of saline-injected group. The results suggest that p16INK4a may be associated with apoptosis of luminal epithelial cells and decidual cells, coordinating decidualization of endometrium and invasion of trophoblastic cells. Thus, we presume that p16INK4a participates in the process of blastocyst implantation in mice.
Asunto(s)
Animales , Femenino , Ratones , Embarazo , Blastocisto , Fisiología , Inhibidor p16 de la Quinasa Dependiente de Ciclina , Fisiología , Implantación del Embrión , Endometrio , Fisiología , Inmunohistoquímica , ARN Mensajero , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The present study was aimed to investigate the expression of tumor suppressor gene PTEN (phosphatase and tensin homolog deleted on chromosome ten) in mouse endometrium during early pregnancy and its possible role during blastocyst implantation. Real-time fluorescent quantitative PCR (FQ-PCR) and immunohistochemical techniques were applied to detect PTEN mRNA and protein expressions in endometrium in un-pregnant and pregnant mice on days 1, 3, 4, 5, 7 of pregnancy, respectively. In addition, PTEN antisense oligonucleotide was injected into the horns of uterus in pregnant mice on day 3 of pregnancy and its effects on blastocyst implantation was detected in vivo. The higher expressions of PTEN mRNA and protein were observed in pregnant mice compared with that in un-pregnant mice, with a steady increasing from day 1 to 7 and reaching the maximal level on day 5 of pregnancy. PTEN antisense oligonucleotide decreased the number of implanted blastocysts compared with saline. The results suggest that PTEN might associate with apoptosis of luminal epithelial and decidual cells, coordinating decidualization of endometrium and invasion of trophoblastic cells. Thus, PTEN may participate in the process of blastocyst implantation in mice.
Asunto(s)
Animales , Femenino , Ratones , Embarazo , Cromosomas , Implantación del Embrión , Endometrio , Metabolismo , Fosfohidrolasa PTEN , Metabolismo , Trofoblastos , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the effects of total saponins of panax ginseng (TSPG) on proliferation and differentiation of human embryonic neural stem cell (NSC) into dopaminergic neuron.</p><p><b>METHOD</b>Isolation, cultivation and identification of human embryonic NSC from cerebral cortex of 7-12 week abortus. By using flow cytometry and MTT assay, the effects of various concentration of TSPG and TSPG cooperating with cytokines( EGF, bFGF) in NSC culture media for 3 days on proliferation of human embryonic NSC has studied. By employing immunocytochemistry assay of the expression of tyrosine hydroxylase (TH), the effect of different dilution of TSPG and TSPG cooperating with IL-1 on induced differentiation of human embryonic NSC into dopaminergic neuron has researched.</p><p><b>RESULT</b>TSPG can significantly promote the proliferation of NSC. When TSPG cooperating with EGF and bFGF, the proliferation of NSC is much stronger than that of only using FGF and bFGF. TSPG also induces NSC to differentiate into dopaminergic neuron, especially when TSPG is cooperating with IL-1.</p><p><b>CONCLUSION</b>TSPG can not only obviously accelerate the proliferation of NSC, but also significantly induce differentiation of NSC into dopaminergic neuron.</p>
Asunto(s)
Humanos , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Dopamina , Metabolismo , Sinergismo Farmacológico , Células Madre Embrionarias , Biología Celular , Metabolismo , Factor de Crecimiento Epidérmico , Farmacología , Factor 2 de Crecimiento de Fibroblastos , Farmacología , Inmunohistoquímica , Interleucina-1 , Farmacología , Neuronas , Biología Celular , Metabolismo , Panax , Química , Plantas Medicinales , Química , Saponinas , Farmacología , Tirosina 3-Monooxigenasa , MetabolismoRESUMEN
To investigate the effects of total saponins of panax ginseng (TSPG) in combination with hematopoietic growth factors (HGF) on proliferation and differentiation of CD34(+) cells ex vivo, the purified CD34(+) cells from cord blood and bone marrow were expanded by various concentrations of TSPG with combination of cytokines in liquid culture systems and the expanded cell number, CD34(+) cell number, CD33(+) cell ratio, the numbers of total CFC and hematopoietic progenitor cell number were detected. The results showed that TSPG (10 - 70 microg/ml) could raise the expanded cell number, CD34(+) cell number, and the numbers of total CFC, TSPG 50 microg/ml was identified as the most potent stimulating concentration, and increased total nucleated cells to (2470.5 +/- 79.96) x 10(3), CFC to (53.96 +/- 4.286) x 100% and CD34(+) cells to (21.86 +/- 3.094) x 100%; TSPG (10 - 50 microg/ml) could raise the colony formation rate of CFU-GM, TSPG (20 microg/ml) induced the best effect on granulocytopoietic differentiation committed of CD34(+) cells. It is concluded that the optimal concentration of TSPG can promote CD34(+) cells to proliferate and differentiate by cooperating with hematopoietic growth factors.
Asunto(s)
Humanos , Antígenos CD34 , Diferenciación Celular , Proliferación Celular , Sinergismo Farmacológico , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Farmacología , Células Madre Hematopoyéticas , Biología Celular , Alergia e Inmunología , Panax , Química , Saponinas , FarmacologíaRESUMEN
The purpose of the present study was to investigate the biological mechanism for modulating granulocytopoiesis by Panax ginseng. The techniques of culture of hematopoietic progenitor cells and hematopoietic stromal cells in vitro, biological assay of hematopoietic growth factor (HGF), immunocytochemistry, in situ hybridization of nucleic acid, immunoprecipitation and Western blot were used to explore the effect of total saponins of Panax ginseng (TSPG) on the expression of human granulocyte-macrophage colony stimulating factor (GM-CSF) and granulocyte-macrophage colony stimulating factor receptor alpha (GM-CSFRalpha). The results indicated that (1) bone marrow stromal cell (BMSC), thymocyte (TC), splenocyte (SC), endothelial cells (EC), and monocyte (MO) conditioned media prepared with TSPG (50 microg/ml) could significantly enhance the proliferation of CFU-GM; (2) the expressions of GM-CSF in protein and mRNA level in BMSC, TC, SC, EC and MO induced by TSPG (50 microg/ml) were much higher than that of the control; (3) the expression of GM-CSFRalpha protein in hematopoietic cells induced by TSPG (50 microg/ml) was stronger than that of the control; (4) TSPG (50 microg/ml) could stimulate the transient tyrosine phosphorylation of GM-CSFR and Shc protein. We speculate that TSPG may directly and/or indirectly promote the stromal cells and lymphocytes to produce GM-CSF and other cytokine and induce bone marrow hematopoietic cells to express GM-CSF receptors (GM-CSFRalpha), leading to the regulation of the GM-CSFR-mediated signals transduction pathway and the proliferation of human CFU-GM.