1.Endoplasmic reticulum stress participates in uric acid-induced phenotypic transformation of renal tubular epithelial cells
Pu WU ; Fei ZHAO ; Dan NIU ; Xinyang WANG ; Yaning HAO
Chinese Journal of Nephrology 2016;32(12):922-927
Objective To explore the effect of endoplasmic reticulum stress (ER stress) in uric acid?induced phenotypic change in renal tubular epithelial cells (HK?2). Methods (1) HK?2 cells were cultured with 0, 75, 150, 225, 300 mg/L uric acid for 24 h in vitro. (2) The cells were divided into normal control group, ER stress inhibitor 4?PBA (5 μmol/L) group, uric acid (150 mg/L) group and 4?PBA+uric acid group for 24 h. Morphological changes of HK?2 cells were observed under inverted microscope. MTT assay was used to detect the proliferation of HK?2 cells treated with 150 mg/L uric acid for 24, 48 and 72 h. The protein expressions of α?smooth muscle actin (α?SMA), vimentin, snail, glucose regulated protein 78 (GRP78) and the phosphorylation of eukaryotic initiation factor 2α(p?eIF2α) in HK?2 cells were measured by Western blotting. Results Compared with the control group, HK?2 cells in uric acid groups (150, 225, 300 mg/L) showed fibroblast?like appearance. The protein expressions of α?SMA, vimentin, snail, GRP78 and p?eIF2α in 150 mg/L and 225 mg/L uric acid groups were higher than those in the control group (all P<0.05). The proliferation of HK?2 cells in 150 mg/L uric acid group was lower than that in control group at 48 and 72 h (all P<0.01). Compared with the uric acid group, the cell morphology in 4?PBA+uric acid group was improved, and the protein expressions ofα?SMA, vimentin, snail, GRP78 and p?eIF2α were decreased (all P<0.05). Conclusions Uric acid may induce the phenotype transformation of renal tubular epithelial cell, and ER stress is involved. 4?PBA may inhibit the uric acid?induced ER stress response and phenotypic transformation, and may be beneficial in attenuating uric acid?induced renal tubular damage.
2.Protection of vitamin C on the cardiac injury induced by nano-titanium dioxide in mice
Junzhi TIAN ; Huicai GUO ; Xiaole YUE ; Yi LIU ; Yue ZHU ; Weiyu WANG ; Yaning WANG ; Rong ZHANG ; Yujie NIU
Chinese Journal of Pharmacology and Toxicology 2014;(2):227-232
OBJECTIVE To observe the protection of vitamin C on the cardiac injury induced by 50 nm titanium dioxide inmice.METHODS Kunming mice were ad mistered by ig of vitamin C 100,200 and 400 mg·kg -1 for 2 d.And then the mice were ad mistered by ig of nano-TiO2 2 g·kg -1 and vitamin C (100.0,200.0 and 400.0 mg·kg -1 )for 3 d,the interval of treatment with nano-TiO2 and vitamin C was 4 h.The mice were scarified 24 h later after the last ad ministration.Electrocardiogra m (ECG)was determinated by physiological recorder.The myocardial enzy mes activities in serum and superoxide dismutase (SOD)and glutathione peroxidase(GSH-Px)activities in serum and myocardial tissue were determinated by bioche mical method.Cometassay was used to detect the DNA da mage of the heart. Heart tissue was used for histopathological exa mination by HE staining.RESULTS Co mpared with the control,ECG showed higher S-T and T-wave a mplitude of nano-TiO2 2 g·kg -1 (P<0.05).The myocar-dial enzy mes activities significantly increased and activities of SOD and GSH-Px significantly decreased in nano-TiO2 group,compared with the control group(P <0.05).Cometassay showed that olive tail mo ment (OTM)was significantly increased after nano-TiO2 2 g·kg -1 ,compared with the control group (P<0.05).The histopathology showed ede ma of myocardial cells,myofibril disorders and increasing infla mmatory cells.Vita min C 100,200 and 400 mg·kg -1 can decrease S-T in ECG,OTM,myocardial enzy mes activities,increase the SOD and GSH-Px activities in serum and myocardial tissue;reduce myocardial hypertrophy and infla mmatory cells.CONCLUSION nano-TiO2 can induce myocardial injury inmice and vitamin C can alleviate the da mage.The mechanism may be associated with the antioxidant ability of vitamin C inmyocardial tissue.
3.Differential Diagnosis Between Subcutaneous Hemangioma and Kaposiform Hemangioendothelioma via Different Ultrasonography-Based Radiomics Models
Yaning NIU ; Yihang YU ; Yubin GONG ; Jian DONG ; Jing ZHAO ; Gang WU
Chinese Journal of Medical Imaging 2024;32(7):721-725
Purpose To identify hemangioma(HE)and Kaposiform hemangioendothelioma(KHE)by constructing two ultrasonography-based radiomics models to evaluate the application value of two models in distinguishing HE from KHE,and to compare the diagnostic efficiency of two models.Materials and Methods A total of 90 lesions of subcutaneous HE or KHE confirmed clinically or pathologically from Henan Provincial People's Hospital from August 2020 to May 2022,were retrospectively analyzed.Imaging features were extracted by using Pyradiomics and screened out by the least absolute shrinkage and selection operator algorithm.Support vector machine and random forest were used to construct the radiomics models.Then the diagnostic efficacy of different models was compared.Results Based on the selected 10 radiomics features,the area under the curve,accuracy,sensitivity,specificity,positive and negative prediction the training group and validation group of the support vector machine model were 0.902(95%CI 0.887-0.917),92.1%,85.0%,92.3%,90.9%,93.5%and 0.827(95%CI 0.787-0.856),85.2%,70.0%,94.1%,90.9%,85.0%,respectively;and those in the training group and validation group of the random forest model were 0.960(95%CI 0.938-0.983),98.4%,96.4%,97.8%,98.1%,97.2%and 0.742(95%CI 0.699-0.785),77.8%,57.1%,82.3%,79.6%,62.5%,respectively.The area under the curve between two models in the training group and validation group was statistically significant(Z=-3.306,-2.009;P<0.05).Conclusion Ultrasonography-based radiomics can distinguish HE from KHE,support vector machine model shows more stable diagnostic performance in small sample data.
4.Bone marrow mesenchymal stem cell-derived exosomal miR-30e-5p ameliorates high-glucose induced renal proximal tubular cell pyroptosis
Jia LÜ ; Yaning HAO ; Xiaopei WANG ; Wanhong LU ; Yali ZHANG ; Dan NIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(6):859-865
【Objective】 To study the effects of miR-30e-5p from bone marrow mesenchymal stem cell-derived exosomes(BMSC-exos) on high glucose (HG)-induced HK-2 cell pyroptosis and explore an alternative strategy to manage diabetic kidney disease (DKD). 【Methods】 BMSC-exos were isolated and internalized into HK-2 cells treated with HG to measure viability and cytotoxicity. The secretion of IL-1β and IL-18 was measured by ELISA. Pyroptosis was assessed by flow cytometry. The levels of miR-30e-5p, IL-1β, and IL-18 were measured. The expression of pyroptosis-associated cytokine proteins was determined. 【Results】 BMSC-exos decreased LDH, IL-1β, and IL-18 secretion and inhibited the expression of the pyroptosis-related factors (IL-1β, caspase-1, GSDMD-N, and NLRP3) in HG-induced HK-2 cells. Moreover, miR-30e-5p depletion in BMSC-exos promoted HK-2 cell pyroptosis. 【Conclusion】 BMSC-derived exosomal miR-30e-5p inhibits caspase-1-mediated pyroptosis in HG-induced HK-2 cells, which might provide a new strategy for treating DKD.
5.Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies.
Shenli ZHANG ; Qianru XU ; Jifei YANG ; Qingmei LI ; Yaning SUN ; Xueyang LI ; Yanan WANG ; Xiangxiang NIU ; Xiaotian QU ; Jinxuan CHEN ; Erqin ZHANG ; Gaiping ZHANG
Chinese Journal of Biotechnology 2022;38(5):1981-1993
The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
Animals
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Antibodies, Viral
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Chickens
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HN Protein/metabolism*
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Newcastle Disease/prevention & control*
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Newcastle disease virus/metabolism*
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Oryza/genetics*