Astragaloside IV alleviates 1-deoxysphinganine-induced mitochondrial dysfunction during the progression of chronic kidney disease through p62-Nrf2 antioxidant pathway.

Introduction: Chronic kidney disease (CKD) can lead to significant elevation of 1-deoxysphingolipids (1-deoxySL). The increase of 1-deoxySL in turn can result in mitochondrial damage and oxidative stress, which can cause further progression of CKD. Methods: This study assessed the therapeutic effect of Astragaloside IV (AST) against 1-deoxySL-induced cytotoxicity in vitro and in rats with CKD. HK-2 cells were exposed to 1-deoxysphinganine (doxSA) or doxSA + AST. doxSA-induced mitochondrial dysfunction and oxidative stress were evaluated by immunostaining, real-time PCR, oxidative stress sensor, and transmission electron microscopy. The potential effects of AST on kidney damage were evaluated in a rat 5/6 nephrectomy (5/6 Nx) model of CKD. Results: The findings of in vitro experiments showed that doxSA induced mitochondrial damage, oxidative stress, and apoptosis. AST markedly reduced the level of mitochondrial reactive oxygen species, lowered apoptosis, and improved mitochondrial function. In addition, exposure to AST significantly induced the phosphorylation of p62 and the nuclear translocation of Nrf2 as well as its downstream anti-oxidant genes. p62 knock-down fully abolished Nrf2 nuclear translocation in cells after AST treatment. However, p62 knock-down did not affect TBHQ-induced Nrf2 nuclear translocation, indicating that AST can ameliorate doxSA-induced oxidative stress through modulation of p62 phosphorylation and Nrf2 nuclear translocation. Conclusion: The findings indicate that AST can activate Nrf2 antioxidant pathway in a p62 dependent manner. The anti-oxidative stress effect and the further mitochondrial protective effect of AST represent a promising therapeutic strategy for the progression of CKD.

The role of CRP and ATG9B expression in clear cell renal cell carcinoma.

The purpose of the study is to investigate the correlation between the expression of C-reactive protein (CRP) and autophagy-related 9B (ATG9B) and pathological features of clear cell renal cell carcinoma (CCRCC) patients. We also intended to explore the effects of manipulated expression of CRP and ATG9B on the apoptosis and cell cycle progression of CCRCC cell line. ATG9B expression in CCRCC tissues and adjacent renal tissues was analyzed by immunohistochemistry (IHC). Gene expression was determined at transcription and translational levels using real-time quantitative PCR (RT-qPCR) and Western blot. The association between CRP/ATG9B expression and clinical-pathological parameters including age, gender, pathological grades, TNM stage and distant metastasis of the patients was assessed by correlation analysis. siRNA and overexpression plasmids construction were used to manipulate the expression of CRP in human CCRCC cell line 786-O. Cell apoptosis and cell cycle progression were determined using flow cytometry (FCM) and Hoechst 33258 staining. CRP expression correlates with ATG9B expression. The expression of CRP and ATG9B are significantly correlated with TNM staging, distant metastasis, and survival time of CCRCC patients. A high-level of CRP indicates a poor overall survival (OS). In addition, CRP expression influences cell cycle and apoptosis of CCRCC cells. The study reveals that CRP might be a CCRCC development promoter. In addition, there is a close relationship between CRP and ATG9B in CCRCC carcinogenesis.

[Screening of Y chromosome microdeletions in infertile males with varicocele].

OBJECTIVE: To investigate the relationship between Y chromosome microdeletions and human spermatogenesis in infertile men with varicocele (VC). METHODS: We divided 174 infertile VC patients into groups A (with azoospermia, n = 47) , B (with severe oligozoospermia, n=57) and C (with mild oligozoospermia, n=70), and enlisted 28 fertile males and 26 fertile females as normal controls. We collected DNA from the peripheral blood, amplified 6 sequence tagged sites in AZFa, AZFb and AZFc using multiplex PCR technique. Then we separated and scanned the amplified products by agarose gel electrophoresis to identify microdeletions and their types in comparison with the controls. RESULTS: Y chromosome microdeletions were observed in 12.64% of the patients (22/174), 11 cases in group A and the other 11 in group B, but none in group C and the normal controls. The differences were statistically significant (P<0.05). In group A, 6 of the microdeletion cases were in the AZFc region, 1 in the AZFa region, 2 in the AZFb region and 2 in both AZFb and AZFc regions, while in group B, 8 cases were in the AZFc region, 2 in the AZFb region and 1 in both AZFb and AZFc regions. CONCLUSION: Infertility is correlated to Y chromosome microdeletions in VC patients. Y chromosome microdeletion screening should be performed for infertile VC patients, especially for those with azoospermia or severe oligozoospermia.

Preparation and study on the novel solid inclusion complex of ciprofloxacin with HP-beta-cyclodextrin.

The interaction of ciprofloxacin with HP-beta-cyclodextrin (HP-beta-CD) has been studied by several analytical techniques, including 1H nuclear magnetic resonance (NMR), 13C NMR, fluorescence spectra, infrared spectroscopy, thermal analyzer and scanning electron microscope. In this paper, solid inclusion complex of ciprofloxacin with HP-beta-CD was synthesized by the coprecipitation method. In addition, the characterization of the inclusion complex has been proved by fluorimetry, infrared, differential scanning calorimetry and one-dimensional (1D), 2D NMR. The experimental results confirmed the existence of 1:1 inclusion complex of ciprofloxacin with HP-beta-CD. The formation constant of complex was determined by fluorescence method and 1H NMR. Spacial configuration of complex has been proposed on two-dimensional NMR technique.

Preparation and study on the solid inclusion complex of sparfloxacin with HP-beta-cyclodextrin.

The interaction of sparfloxacin with HP-beta-cyclodextrin (HP-beta-CD) has been studied by several analytical techniques, including 1H NMR, fluorescence spectroscopy, infrared spectroscopy, thermal analysis and scanning electron microscopy. In this paper, solid inclusion complex of sparfloxacin with HP-beta-CD was synthesized by the coprecipitation method. In addition, the characterization of the inclusion complex has been proved by fluorimetry, infrared, differential scanning calorimetry and 1D, 2D NMR. The experimental results confirmed the existence of 1:1 inclusion complex of sparfloxacin with HP-beta-CD. The formation constant of complex was determined by the fluorescence method and 1H NMR. Spacial configuration of complex has been proposed on 2D NMR technique.