Long-term hypertension causes excessive vascular remodeling and leads to adverse cardiovascular events. Balance of ubiquitination and deubiquitination has been linked to several chronic conditions, including pathological vascular remodeling. In this study, we discovered that the expression of ubiquitin-specific protease 25 (USP25) is significantly up-regulated in angiotensin II (Ang II)-challenged mouse aorta. Knockout of Usp25 augments Ang II-induced vascular injury such as fibrosis and endothelial to mesenchymal transition (EndMT). Mechanistically, we found that USP25 interacts directly with Forkhead box O3 (FOXO3) and removes the K63-linked ubiquitin chain on the K258 site of FOXO3. We also showed that this USP25-mediated deubiquitination of FOXO3 increases its binding to light chain 3 beta isoform and autophagosomic-lysosomal degradation of FOXO3. In addition, we further validated the biological function of USP25 by overexpressing USP25 in the mouse aorta with AAV9 vectors. Our studies identified FOXO3 as a new substrate of USP25 and showed that USP25 may be a potential therapeutic target for excessive vascular remodeling-associated diseases.

Doxorubicin (Dox) is an anthracycline drug widely applied in various malignancies. However, the fatal cardiotoxicity induced by Dox limits its clinical application. Post-transcriptional protein modification via ubiquitination/deubiquitination in cardiomyocytes mediates the pathophysiological process in Dox-induced cardiotoxicity (DIC). In this study, we aimed to clarify the regulatory role and mechanism of a deubiquitinating enzyme, ubiquitin-specific peptidase 13 (USP13), in DIC. RNA-seq analysis and experimental examinations identified that cardiomyocyte-derived USP13 positively correlated with DIC. Mice with cardiac-specific deletion of USP13 were subjected to Dox modeling. Adeno-associated virus serotype 9 (AAV9) carrying cTNT promoter was constructed to overexpress USP13 in mouse heart tissues. Cardiomyocyte-specific knockout of USP13 exacerbated DIC, while its overexpression mitigated DIC in mice. Mechanistically, USP13 deubiquitinates the stimulator of interferon genes (STING) and promotes the autolysosome-related degradation of STING, subsequently alleviating cardiomyocyte inflammation and death. Our study suggests that USP13 serves a cardioprotective role in DIC and indicates USP13 as a potential therapeutic target for DIC treatment.

OBJECTIVE：To establish a method for simultaneous determination of gallic acid ，protocatechuic acid ，mangiferin， homomangiferin，hyperoside and isoquercetin in Mangguo zhike tablets. METHODS ：HPLC was adopted. The determination was performed on Phenomenex Gemini C 18 column with mobile phase consisted of 0.1％ phosphoric acid water solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 258 nm. The column temperature was set at 30 ℃，and sample size was 5 μL. Gallic acid was used as the internal substance，and the correction factors of gallic acid ， protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were calculated ；the results of quantitative analysis of multi-components by single marker method （QAMS） were compared with those of external standard method （ESM）. RESULTS：The linear range of gallic acid ，protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were 45.75-183 0 μg/mL（r＝0.999 9），2.525-101.0 μg/mL（r＝0.999 9），65.33-261 3 μg/mL（r＝0.999 6），9.058-362.3 μg/mL（r＝ 0.999 9），3.885-155.4 μg/mL（r＝0.999 9），1.870-74.8 μg/mL（r＝0.999 9），respectively. The limits of quantification were 0.571， 0.643，1.053，0.854，0.830，1.500 μg/mL，respectively. The limits of detection were 0.171，0.193，0.316，0.256，0.249，0.450 μg/mL， respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％ . The average recoveries were 98.8％-101.9％（RSD＝1.3％ ，n＝6），94.3％-101.5％（RSD＝3.3％，n＝6），97.9％-100.5％（RSD＝0.9％，n＝6），98.2％- 101.6％ （RSD＝1.2％，n＝6），102.3％-106.1％（RSD＝1.3％，n＝6）， 96.6％ -99.3％（RSD＝1.0％ ，n＝6），respectively. RCFs of 73） protocate-chuic acid ， mangiferin， homomangiferin，hyperoside and isoquercetin were 0.568 3，0.500 3，0.687 6， 0.939 1 and 0.826 3，using gallic acid as internal substance . The content ranges of protocatechuic acid and other 4 com- ponents measured by QAMS were 0.197-0.440，3.262-11.250， 0.201-1.196，0.168-0.381，0.115-0.293 mg/tablet，respectively. The content ranges measured by ESM were 0.198-0.441，3.239-11.570，0.206-1.194，0.171-0.380，0.119-0.298 mg/tablet， respectively. By comparing the content determination results by QAMS and ESM ，the relative errors were －3.80％-0.74％，and there was no statistical significance （P＞0.05）. CONCLUSIONS ：The established QAMS method is accurate ，reliable and repeatable，and can be used for content determination of 6 medicinal components in Mangguo zhike tablets.

Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Humans ; Lymphoma, Follicular ; drug therapy ; Rituximab

OBJECTIVEA prospective, multicenter and non-interventional prospective study was conducted to evaluate the clinical features of rituximab combined with chemotherapy (R-Chemo) as first-line treatment on newly diagnosed Chinese patients with diffuse large B-cell lymphoma (DLBCL).

METHODSThis was a single arm, prospective, observational multicenter and phase IV clinical trial for 279 patients, who were newly diagnosed as CD20-positive DLBCL from 24 medical centers in China 2011 and 2012, no special exclusion criteria were used. All patients received rituximab based R-Chemo regimes, such as R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisolone) and other regimes as the first-line treatment. The treatment strategies were determined by physicians and patients without detailed description for treatment course, dose, interval time and examination. Clinical response and safety of all patients were investigated in 120 days after completion of last dose of rituximab.

RESULTSOf 279 patients, 258 with stage I-IV who received at least 1 cycle of rituximab treatment and completed at least one time of tumor assessment were enrolled into intention-to-treat analysis, including 148 male and 110 female. The median age of all patients was 57.2(12.8-88.4) years. ECOG performance statuses of 0 or 1 were observed in 91.1% of patients, international prognostic index levels in the low-risk and low-middle-risk groups in 76.4% of patients, the tumor diameters smaller than 7.5 cm in 69.0% of patients. All patients received 6 median cycles of R-Chemo treatment every 24.4 days. R-CHOP treatment was shown to improve the clinical response with overall response rates of 94.2%. Common adverse events included anemia, marrow failure, leukopenia, thrombocytopenia, digestive diseases, infection and liver toxicity. All adverse events are manageable.

CONCLUSIONNon-interventional clinical trial of R-Chemo remains the standard first-line treatment for newly diagnosed patients with DLBCL in real clinical practice, which is consistent with international treatment recommendations for DLBCL patients. R-Chemo can provide the clinical evidence and benefit as the first-line standard treatment for Chinese patients with DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; Male ; Middle Aged ; Prospective Studies ; Rituximab ; Treatment Outcome