Objective: To investigate the electrochemiluminescence (ECL) behavior of cloperastine hydrochloride. Methods: ECL intensity of tris (2,2′-bipyridyl) rutheniumo(II) was enhanced, the method for the determination of cloperastine hydrochloride was established using capillary electrophoresis (CE) coupled with electrochemilumolinescence (ECL) detection. Results: Under the optimum conditions, ECL intensity varied linearly with cloperastine hydrochloride concentration from 7.0 × 10-6g/mL to 1.0 × 10-4g/mL. The detection limit (S/N=3) was 8.05 × 10-7 g/mL. The relative standard deviation of the ECL intensity and the migration time for 11 consecutive iajections of 1.0 × 10-5 g/mL cloperastine hydrochloride was 2.9% and 1.5%, respectively. This method was successfully applied to cloperastine hydrochloride tablet determination. Conclusion: The method has been established, validated and applied for determination of cloperastine hydrochloride.

Objective: To investigate the electrochemiluminescence (ECL) behavior of cloperastine hydrochloride. Methods: ECL intensity of tris (2,2′-bipyridyl) rutheniumo(II) was enhanced, the method for the determination of cloperastine hydrochloride was established using capillary electrophoresis (CE) coupled with electrochemilumolinescence (ECL) detection. Results: Under the optimum conditions, ECL intensity varied linearly with cloperastine hydrochloride concentration from 7.0 × 10-6g/mL to 1.0 × 10-4g/mL. The detection limit (S/N=3) was 8.05 × 10-7 g/mL. The relative standard deviation of the ECL intensity and the migration time for 11 consecutive iajections of 1.0 × 10-5 g/mL cloperastine hydrochloride was 2.9% and 1.5%, respectively. This method was successfully applied to cloperastine hydrochloride tablet determination. Conclusion: The method has been established, validated and applied for determination of cloperastine hydrochloride.

High expression of Fightless I (FLII) is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins Importin β and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them into the Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.
Humans ; Microfilament Proteins ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Recombinant Fusion Proteins ; metabolism ; beta Karyopherins ; metabolism

Objective:To discuss the effect of curcumin on ameliorating doxorubicin(DOX)-induced cytotoxicity in the H9c2 cardiomyocytes,and to clarify its mechanism.Methods:The H9c2 cardiomyocytes were treated with DOX to establish the cardiotoxicity model.The H9c2 cells were divided into normal group,DOX group,DOX+curcumin group,and DOX+curcumin+silent information regulator 3(SIRT3)inhibitor-3(3-TYP)group.After 24 h,the morphology of the cells in various groups were observed;CCK-8 method was used to detect the viabilities of the cells in various groups;TUNEL staining was used to detect the apoptotic rates of the cells in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the activities of catalase(CAT)and superoxide dismutase(SOD)and the levels of malondialdehyde(MDA)in the cells in various groups;2,7-dichlorofluorescein diacetate(DCFH-DA)staining was used to detect the levels of reactive oxygen species(ROS)in the cells in various groups;Western blotting method was used to detect the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),nicotinamide adenine dinucleotide phosphate(NADPH)oxidase 2(NOX2),NADPH oxidase 4(NOX4),SIRT3,acetylated superoxide dismutase 2(Ac-SOD2),and superoxide dismutase 2(SOD2)proteins in the cells in various groups.Results:Compared with normal group,the H9c2 cells in DOX group exhibited swelling,the activity of the cells was decreased(P＜0.05),the CAT and SOD activities were decreased(P＜0.05),the MDA and ROS levels were increased(P＜0.05),the expression levels of NOX2,and NOX4 proteins were increased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were decreased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were increased(P＜0.05).Compared with DOX group,the H9c2 cells in DOX+curcumin group showed improved morphology,the activity of the cells was increased(P＜0.05),the CAT and SOD activities were increased(P＜0.05),the MDA and ROS levels were decreased(P＜0.05),the expression levels of NOX2,and NOX4 proteins were decreased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were increased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were decreased(P＜0.05).Compared with DOX+curcumin group,the cells in DOX+curcumin+3-TYP group exhibited swelling,the activity of the cells was decreased(P＜0.05),the apoptotic rate of the cells was significantly increased(P＜0.05),the CAT and SOD activities were decreased(P＜0.05),the MDA and ROS levels were significantly increased(P＜0.05),the expression levels of NOX2 and NOX4 proteins were increased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were decreased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were increased(P＜0.05).Conclusion:Curcumin suppresses the oxidative stress and apoptosis by activating the SIRT3/SOD2 signaling pathway,improving the cell activity and alleviating the DOX-induced cytotoxicity in the H9c2 cells.

OBJECTIVES: To explore the effect of resveratrol (Res) on Kawasaki disease (KD)-induced myocardial injury and to evaluate its effect on apoptosis and autophagy. METHODS: Forty-eight juvenile male Sprague Dawley rats were randomly divided into a control group, a Res group, a lactobacillus casei cell wall extract (LCWE)-induced Kawasaki disease group (KD group), and a LCWE-induced Kawasaki disease + Res treatment group (Res+KD group). The control group was intraperitoneally injected with saline. The Res group was intraperitoneally injected with resveratrol (100 mg/kg). The KD group was intraperitoneally injected with 0.5 mL LCWE (1 mg/mL). The Res+KD group was intraperitoneally injected with 0.5 mL LCWE (1 mg/mL) and resveratrol (100 mg/kg). After 4 weeks, the left ventricular ejection fraction (LVEF) and short axis shortening rate (LVFS) were detected by echocardiography. The apoptotic rate was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. The levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), microtubule-associated protein 1 light chain 3β (LC3B), Beclin-1, autophagy related 5 (Atg5) and sequestosome-1 (p62) were detected by Western blotting. The formation of autophagosome was observed under transmission electron microscope. RESULTS: There was no significant difference in the above-mentioned indexes between the control group and the Res group (all CONCLUSIONS Res can attenuate the KD-induced myocardial injury via inhibiting the apoptosis and autophagy.
Animals ; Apoptosis ; Autophagy ; Male ; Mucocutaneous Lymph Node Syndrome/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Resveratrol/therapeutic use* ; Stroke Volume ; Ventricular Function, Left