OBJECTIVETo Study the effect of anti-aging and its mechanism of total saponins of Wu-He Dipsacus asper on skin of mice-aging model.

METHODSForty-eight mice were randomly divided into blank control group, model group, low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group( n = 8) . The mouse model of skin aging was established by nape subcutaneous injection of 5% D-galactose (0.025 mL/(g · d)), the mouse of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group were administered with total saponins of Wu-He Dipsacus asper (50 ml/(kg · d), 100 mL/(kg · d), 200 mL/(kg · d)), the mice of the positive control group were administered with vitamin E(50 mg/(kg · d)) for 42 d. The content of hydroxyproline (HYP) and lipofuscin (LF) were measured in skin of each group mice, the activity of catalase (CAT) glutathione peroxidase ( GSH-Px) superoxide dismutase (SOD) and the content of malondi- aldehyde (MDA) were determined in serum and skin of each group mice.

RESULTSCompared with blank control group, the content of HYP decreased significantly and the content of LF increased significantly in skin, the activities of CAT, GSH-Px and SOD decreased significantly and the content of MDA increased significantly in serum and skin of model group; Compared with model group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the con- tent of MDA decreased significantly in serum and skin of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group; Compared with low-Dipsacus group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the content of MDA decreased significantly in serum and skin of high-Dipsacus group and positive control group; The activity of SOD in serum and skin had a significant positive correlation with the content of HYP, and a significant negative correlation with LF in skin.

CONCLUSIONTotal saponins of Wu-He Dipsacus asper have obvious effect of anti-agng on skin of mouse-aging model , its mechanism is closely related to oxidative damage.

Animals ; Dipsacaceae ; chemistry ; Disease Models, Animal ; Mice ; Oxidative Stress ; Saponins ; pharmacology ; Skin Aging ; drug effects

To study the antiarrhythmic effect of the newly developed alpha/beta-blocker TJ0711, a variety of animal models of arrhythmia were induced by CaCl2, ouabain and ischemia/reperfusion. Glass microelectrode technique was used to observe action potentials of right ventricular papillary muscle of guinea pig. The onset time of arrhythmia induced by CaCl2 was significantly prolonged by TJ0711 at 0.75, 1.5 and 3 mg x kg(-1) doses. TJ0711 (1.5 and 3 mg x kg(-1)) can significantly shorten the ventricular tachycardia (VT) and ventricular fibrillation (VF) duration, the incidence of VF and mortality were significantly reduced. On ischemia-reperfusion-induced arrhythmic model, TJ0711 (0.25, 0.5, 1 and 2 mg x kg(-1)) can significantly reduce the ventricular premature contraction (PVC), VT, VF incidence, mortality, arrhythmia score with a dose-dependent manner. At the same time, rats serum lactate dehydrogenase (LDH) and creatine kinase (CK) activities decreased significantly by TJ0711 (1 and 2 mg x kg(-1)). Ouabain could cause arrhythmia in guinea pigs, when TJ0711 (0.375, 0.75, 1.5 and 3 mg x kg(-1)) was given, the doses of ouabain inducing a variety of arrhythmia PVC, VT, VF, cardiac arrest (CA) were significantly increased with a dose-dependent manner. In the TJ0711 0.1-30 micromol x L(-1) concentration range, guinea pig right ventricular papillary muscle action potential RP (rest potential), APA (action potential amplitude) and V(max) (maximum velocity of depolarization) were not significantly affected. APD20, APD50 and APD90 had a shortening trend but no statistical difference with the increase of TJ0711 concentration. TJ0711 has antiarrhythmic effect on the sympathetic nerve excitement and myocardial cell high calcium animal arrhythmia model. Myocardial action potential zero phase conduction velocity and resting membrane potential were not inhibited by TJ0711. APD20, APD50 and APD90 were shortened by TJ0711 at high concentration. Its antiarrhythmic action mechanism may be besides the action of blocking beta1 receptor, may also have a strong selective blocking action on alpha1 receptor and reducing intracellular calcium concentration.
Action Potentials ; drug effects ; Adrenergic alpha-Antagonists ; administration & dosage ; pharmacology ; Adrenergic beta-Antagonists ; administration & dosage ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; administration & dosage ; pharmacology ; Arrhythmias, Cardiac ; blood ; chemically induced ; etiology ; pathology ; physiopathology ; Calcium Chloride ; Creatine Kinase ; blood ; Dose-Response Relationship, Drug ; Female ; Guinea Pigs ; Heart Ventricles ; cytology ; Lactate Dehydrogenases ; blood ; Male ; Myocardial Reperfusion Injury ; complications ; Myocytes, Cardiac ; drug effects ; physiology ; Ouabain ; Papillary Muscles ; cytology ; Phenoxypropanolamines ; administration & dosage ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Aim To investigate the combined pharmacokinetic-pharmacodynamic(PK-PD) model of irbesartan in healthy Chinese male volunteers. Methods Eighteen healthy male volunteers received 300 mg irbesartan tablet orally. The plasma drug concentration (C_p) was determined by HPLC and pharmacologic effects, including SBP, DBP and HR, were measured simultaneously. The pharmakinetic and PK-PD model parameters were calculated. Results The pharmacokinetic profiles were best fitted a two-compartment open model. There was a hysteresis loop between effects and plasma concentrations. The relationship between effects and effect compartment concentrations of drugs could be represented by the Sigmoid-E_ max model. Conclusion We successfully eatablished the PK-PD model of irbesartan in healthy male volunteers. These findings may provide a more rational basis for patient-specific dosage individualization.

To study the antiarrhythmic effect of the newly developed alpha/beta-blocker TJ0711, a variety of animal models of arrhythmia were induced by CaCl2, ouabain and ischemia/reperfusion. Glass microelectrode technique was used to observe action potentials of right ventricular papillary muscle of guinea pig. The onset time of arrhythmia induced by CaCl2 was significantly prolonged by TJ0711 at 0.75, 1.5 and 3 mg x kg(-1) doses. TJ0711 (1.5 and 3 mg x kg(-1)) can significantly shorten the ventricular tachycardia (VT) and ventricular fibrillation (VF) duration, the incidence of VF and mortality were significantly reduced. On ischemia-reperfusion-induced arrhythmic model, TJ0711 (0.25, 0.5, 1 and 2 mg x kg(-1)) can significantly reduce the ventricular premature contraction (PVC), VT, VF incidence, mortality, arrhythmia score with a dose-dependent manner. At the same time, rats serum lactate dehydrogenase (LDH) and creatine kinase (CK) activities decreased significantly by TJ0711 (1 and 2 mg x kg(-1)). Ouabain could cause arrhythmia in guinea pigs, when TJ0711 (0.375, 0.75, 1.5 and 3 mg x kg(-1)) was given, the doses of ouabain inducing a variety of arrhythmia PVC, VT, VF, cardiac arrest (CA) were significantly increased with a dose-dependent manner. In the TJ0711 0.1-30 micromol x L(-1) concentration range, guinea pig right ventricular papillary muscle action potential RP (rest potential), APA (action potential amplitude) and V(max) (maximum velocity of depolarization) were not significantly affected. APD20, APD50 and APD90 had a shortening trend but no statistical difference with the increase of TJ0711 concentration. TJ0711 has antiarrhythmic effect on the sympathetic nerve excitement and myocardial cell high calcium animal arrhythmia model. Myocardial action potential zero phase conduction velocity and resting membrane potential were not inhibited by TJ0711. APD20, APD50 and APD90 were shortened by TJ0711 at high concentration. Its antiarrhythmic action mechanism may be besides the action of blocking beta1 receptor, may also have a strong selective blocking action on alpha1 receptor and reducing intracellular calcium concentration.

Objective To investigate the changes and roles of Fc gamma receptors (FcγR) expressed on monocytes in the immune pathogenesis of Henoch-Schonlein purpura(HSP).Methods Thirty children of HSP and 15 health controls were enrolled in this study.The expressions of FcγR Ⅰ and FcγRⅢ on monocytes were determined by flow cytometry,and real-time PCR was performed to detect the transcription levels of FcγR Ⅱ a,FcγR Ⅱ b,cytokines(IL-1 β,IL-6,TNF-α,IFN-α),chemokine(IP-10,RANTES,iNOS),and BLyS/April in monocytes isolated by microbeads.The plasma concentrations of IL-4,IL-10 and TNF-α were analyzed by enzyme-linked immunosorbent assay (ELISA).Results (1) The expressions of FcγR Ⅰ and FcγR Ⅲ on monocytes in patients with HSP were significantly up-regulated compared with healthy controls.Transcription level of FcγR Ⅱ a on monocytes in patients with acute HSP was found to be higher than that in healthy controls while the inhibitory FcγR Ⅱ] b mRNA was significant down-regulated(P＜0.05),which resulted in a higher ratio of FcR Ⅱ a/Ⅱ b in patients with acute HSP.(2) The expressions of cytokine/chemokines factor such as IL-1 β,IL-6,TNF-α,IP-10,RANTES,and iNOS in patients with HSP was detected to be higher than those in healthy controls(P＜0.05).In addition,expression levels of BLyS/April were up-regulated during acute HSP(P＜0.05),the positive correlations were observed between the FcγR Ⅱ a/FcγR Ⅱ b and the cytokine/chemokines factor in monocytes(P＜0.05).(3) Plasma concentrations of IL-4,IL-10 and TNF-α were significantly elevated during acute HSP(P＜0.05),and a negative correlation was observed between concentrations of TNF-α and the mRNA level of FcγR Ⅱb in monocytes.Conclusion The abnormal expression of the cytokines and the imbalance of stimulatory and inhibitory FcγR of monocyte in acute HSP.

Objective To investigate the correlation between deep vein thrombosis(DVT) and clinical laboratory tests in the regulation of hemostasis. Methods Endothelin-1 (ET-1), thrombomodulin (TM), P-selectin, prothrombin fragment _ 1+2 (F_ 1+2 ), thrombin-antithrombin complex (TAT), thrombus precursor protein (TpP) and D-Dimer (D-D) were measured in DVT patients (n=72) and normal individuals (n=20) with ELISA method. The sensitivity, specificity, negative predictive value (NPV), positive predictive value (PPV) and accuracy were evaluated for each of the items, and the area of the underlying receiver operating characteristic curve (AUC ROC ) was used to appraise diagnostic efficiency. Results Except ET-1, there were significant differences between the DVT group and normal control group in the other items. The AUC ROC of P-selectin, F_ 1+2 , TpP and D-D was bigger than that of ET-1, TM and TAT (P0.05). The TpP had the highest sensitivity, NPV and accuracy, and the D-D had the highest specificity and PPV. Conclusion The increase of D-D and TpP could reflect the occurrence of continuous coagulation and fibrolysis respectively. Thus the combination of measuring TpP and D-D could provide better laboratory evidence for the diagnosis of patients with DVT.

Objective To investigate the effects of the intravenous immunoglobulin (IVIG) on the expression of FcγR Ⅱ b and Toll-like receptor 4 (TLR4) in children with Kawasaki disease (KD).Methods 25 children with KD and 15 healthy controls were enrolled in this study.The levels of TLR4 expression on monocytes were assessed by flow cytometry.Real-time PCR was performed to detect the transcription levels of FcγR Ⅱ b,TLR4 signaling molecules(MyD88,TRAF-6,TAK1) and cytokines(IL-1β,IL-6,TNF-α) in monocytes.The concentrations of TNF-α in plasma was determined by enzyme-linked immunosorbent assay (ELISA).Results (1) Compared with healthy controls,the expression of TLR4 and the signaling molecules in the patients were significantly up-regulated but decreased after treatment with IVIG (P＜0.05).(2)The levels of FcγR Ⅱb expression from the patients were higher than those from healthy controls,and decreased after treatment with IVIG(P＜0.05).(3) The levels of cytokine factor expression such as IL-1β,IL-6 and TNF-α in the patients were found to be higher than those in healthy controls (P＜0.05) ; plasma concentrations of TNF-α were significantly elevated during acute KD (P＜0.05).(4)There were significantly correlations to follow between FcγR Ⅱb and the levels of cytokine,TLR4 expression in monocytes (P ＜0.05).Conclusion IVIG can up regulate FcγR Ⅱb expression and down regulate TLR4 expression which might inhibit the inflammatory response.

Objective To investigate the effects of sIL-2R in plasma on regulatory T cells (Treg) in children with Kawasaki disease ( KD ) .Methods Thirty-three children with KD and fourteen age-matched healthy children were enrolled in this study .The proportions of CD4+CD25+Foxp3+Treg cells in pe-ripheral blood and mean fluorescence intensity (MFI) of phosphorylated-STAT5 (pSTAT5) protein in CD4+CD25+T cells were analyzed by flow cytometry .The concentrations of sIL-2R, IL-2, IL-7 and IL-15 in plas-ma were measured by cytometric bead array ( CBA ) .Real-time PCR was performed to detect the gene ex-pressions of Foxp3, GITR, CTLA4, IL-2Rα, IL-2Rβand IL-2Rγat mRNA level as well as the expressions of IL-17A and ROR-γt at mRNA level in CD4+CD25-T cells.Results (1) Compared with the control group, the proportions of CD4+CD25+Foxp3+Treg cells in peripheral blood from patients with KD and the expressions of associated factors including Foxp 3, GITR and CTLA-4 at mRNA level were significantly down-regulated (P<0.05).However,the expressions of IL-17A and ROR-γt at mRNA level in Th17 cells were markedly up-regulated (P<0.05), which could be recovered to some extent after treatment with IVIG (P<0.05).(2)The expressions of pSTAT5 protein in CD4+CD25+T cells from patients with acute KD were sig-nificantly decreased (P<00.5 ), but increased with IVIG intervention (P<0.05).(3)The concentrations of sIL-2R in plasma were elevated during acute KD (P<0.05), but decreased after treatment with IVIG (P<0.05).Moreover, KD patients with coronary artery lesion ( KD-CAL+) presented a high level of sIL-2R than those without coronary artery lesion (KD-CAL-) (P<0.05), but there was no significant difference in the concentrations of IL-2, IL-7 and IL-15 in plasma between two groups (P>0.05).(4)The expressions of IL-2Rαand IL-2Rβat mRNA level in CD4+CD25+T cells from patients with acute KD were lower than those of the healthy subjects (P<0.05), but up-regulated to some extent with IVIG treatment (P<0.05).There was no significant change in the expression of IL-2Rγat mRNA level (P>0.05).The concentrations of sIL-2R in plasma were negatively correlated with the expressions of IL-2Rβand Foxp3 at mRNA level and pSTAT5 at protein level (P<0.05).The expression of pSTAT5 protein had positive correlation with the ex-pression of Foxp3 at mRNA level (P<0.05).Conclusion Aberrant IL-2/STAT5 signaling pathway media-ted by significantly increased concentration of sIL-2R in plasma might be one of the factors leading to down-regulation of Treg cells in patients with acute KD .

Objective To detect the PML-RARα fusion gene expression of APL patients with FISH and fluorescent quantitative PCR and discuss the sensitivity and specificity of two techniques. Methods The detection of the PML-RARα fusion gene expression of 75 APL patients with FISH and fluorescent quantitative PCR were carried out simultaneously. Results Eighty eight patients of primary and relapse or remission phases were examinated and total conformity rate was 96.59 %. Fourteen primary patients were detected and conformity rate was 100 %. Seventy four relapse or remission patients were detected and conformity rate was 95.95 %. Stem cell essays were detected for six times and conformity rate was 100 %. Conclusion The sensitivity of FISH and fluorescent quantitative PCR is identical for primary APL patients and FISH is more sensitive. But the sensitivity of FISH is weaker than that of fluorescent quantitative PCR for detection of residue disease.

Objective To screen the genes related with signal transducers and activators of transcription 3 (STAT3) regulating pancreatic cancer invasion and metastasis by gene chips.Methods Human pancreatic cancer cell line SW1990 stably expressing low level of Stat3 was established by lentivirus transfection,while cells transfected with mock plasmid and cells without transfection served as control groups.The differences of invasion and metastasis related genes expression among the three groups were screened by gene chips.STAT3 mRNA and protein expression was measured by real-time PCR and Western blot.Three differentially expressed genes (MMP-7,IL-1β and IgTα7) were verified.ResultsThe expression level of STAT3 mRNA was 0.391 ± 0.037 after pancreatic cancer SW1990 cell trarsfected with STAT3 targeted lentivirus,which was significantly lower than those in mock plasmid group (1.002 ± 0.015) and nontransfected group ( 1.206 ± 0.042,P ＜ 0.05 ) ; the expression level of STAT3 protein was 182.38 ± 65.32,which was significantly lower than those in mock plasmid group (223.40 ±58.40) and non-transfected group (212.33 ±53.69).Eight invasion and metastasis related genes of SW1990 lowly expressing Stat3 were upregulated,while 3 genes were down-regulated.By verification,the mRNA level of MMP-7 and IL-1β were lower than in control group transfected with mook plassmid(0.287 ± 0.115 vs 1.010 ± 0.124,t =19.45,P =0.000;0.490 ± 0.10 vs 1.002 ± 0.002,t =13.83,P =0.000),but the mRNA level of IgTα7 was not decreased (1.173 ±0.280 vs 0.998 ±0.003,t =4.236,P =0.094).Meanwhile,the protein level of MMP-7 was significantly down-regulated when Stat3 was knocked down.ConclusionsStat3 causes changes of expressions of many invasion and metastasis-related genes of SW1990,and MMP-7 may be the main target gene regulated by Stat3.