1. Mechanism of hydroxysafflower yellow A in the regulation of vascular smooth muscle cell calcification
Yiran HAN ; Tianhua XU ; Xiaobo QIU ; Zitong SHENG ; Pengzhi WAN ; Yue LI ; Li YAO
Chinese Journal of Nephrology 2019;35(11):848-855
Objective:
To investigate the role and mechanism of Hydroxysafflor yellow A (HSYA) in the calcification of vascular smooth muscle cells (VSMC) induced by β-glycerol phosphate (β-GP).
Methods:
VSMC were cultured with 10% fetal bovine serum+1% double anti-high glucose DMEM medium at 37℃ and 5%CO2 incubator, and were subcultured according to cell growth density at 1∶4 ratio. The cells were divided into three groups: control group (NC), high-phosphate-induced calcification (HP) group, and HSYA intervention (HSYA) group. The Calcium deposition amount was measured by alizarin red staining and calcium determination kit. The expressions of ALP, RUNX2, RANKL, α-SMA and inflammation indicators TLR4, TNF-α, IL-8 were detected by Western blotting method; Western blotting was also used to detect calcification index alkaline phosphatase (ALP) and Runt-related transcription factor 2 (RUNX2). Nuclear factor kappa B receptor activating factor ligand(RANKL), α-smooth muscle actin (α-SMA), and the expressions of TLR4/NF-κB pathway and inflammatory response-related indicators Toll-like receptor 4 (TLR4), interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-α). The nuclear protein and cytoplasmic proteins were respectively extracted. The expressions of p65 in nucleus and cytoplasm, as well as the expressions of p65 and phosphorylated p65 in total proteins were detected by Western blotting method. Superoxide dismutase (SOD) and malondialdehyde (MDA) kit were used to detect the content of antioxidant enzymes and oxidation end products in cells.
Results:
Western blotting showed that the expressions of ALP, RUNX2 and RANKL in HSYA group were significantly lower than that in HP group. The expression of α-SMA was increased than that of HP group (all
2.Cordyceps sinensis alleviates β-glycerophosphate-induced vascular smooth muscle call calcification through promoting autophagy
Xiaobo QIU ; Zitong SHENG ; Yiran HAN ; Tianhua XU ; Binyao TIAN ; Fei ZHAO ; Yanqiu YU ; Li YAO
Chinese Journal of Nephrology 2019;35(2):119-126
Objective To investigate the influence mechanism of Cordyceps sinensis (CS) on β-glycerophosphate-induced vascular smooth muscle cell (VSMC) calcification.Methods The effect of CS on VSMC cell viability was detected by CCK-8.The cellular models of rat VSMC calcification were established by treating with β-glycerophosphate (β-GP,10 mmol/L);then CS (10 mg/L),autophagy inhibitor 3-methyladenine (3-MA,5 mmol/L),and AMPK inhibitor compound C (CC,10 μmol/L) were added to the cell cultures.There were a total of 5 experiment groups:VSMC cultured in normal medium (Control),VSMC treated with β-GP,VSMC treated with β-GP and CS,VSMC treated with 3-MA,β-GP and CS,and VSMC treated with CC,β-GP and CS.The calcium nodules and calcium content were examined with alizarin red S staining and the O-cresolphthaleincomplexone method,respectively.The autophagosomes within the VSMC were observed using transmission electron microscope (TEM).Immunofluorescence showed the accumulation of microtubule-associated protein 1 light chain 3 (LC3) puncta.In addition,levels of osteogenic related proteins,autophagy related proteins,and AMPK/mTOR pathway related proteins were evaluated by Western blotting.Results CS increased the number of autophagosomes and the accumulation of LC3 puncta within VSMC.It also upregulated the protein levels of LC3 Ⅱ/LC3 Ⅰ,beclin1,α-SMA,and p-AMPK;whereas,the protein levels of Runx2 and p-mTOR,as well as calcium nodules and calcium content were reduced (all P < 0.01).When the cells were pretreated with 3-MA before treating with β-GP and CS,the autophagosomes,accumulation of LC3 puncta,and protein levels of LC3 Ⅱ/LC3 Ⅰ,beclinl,and α-SMA were decreased (all P < 0.01);however,the protein level of Runx2,and the calcium nodules and calcium content were increased (all P < 0.01).Nevertheless,when the cells were pretreated with CC before giving β-GP and CS,the autophagosomes,the accumulation of LC3 puncta,and the expression levels of p-AMPK,LC3 Ⅱ/LC3 Ⅰ,beclin1,and α-SMA were significantly down-regulated (all P < 0.01);whereas,the expression levels of Runx2 and p-mTOR,as well as calcium nodules and calcium content were increased (all P < 0.01).Conclusions CS can effectively alleviate β-GP-induced VSMC calcification,which may be due to the activation of autophagy by AMPK/mTOR signaling pathway.
3.Discussion on the standard of clinical genetic testing report and the consensus of gene testing industry.
Hui HUANG ; pengzhiyu@bgi.com. ; Yiping SHEN ; Weihong GU ; Wei WANG ; Yiming WANG ; Ming QI ; Jun SHEN ; Zhengqing QIU ; Shihui YU ; Zaiwei ZHOU ; Baixue CHEN ; Lei CHEN ; Yundi CHEN ; Huanhuan CUI ; Juan DU ; Yong GAO ; Yiran GUO ; Chanjuan HU ; Liang HU ; Yi HUANG ; Peipei LI ; Xiaorong LI ; Xiurong LI ; Yaping LIU ; Jie LU ; Duan MA ; Yongyi MA ; Mei PENG ; Fang SONG ; Hongye SUN ; Liang WANG ; Dawei WANG ; Jingmin WANG ; Ling WANG ; Zhengyuan WANG ; Zhinong WANG ; Jihong WU ; Jing WU ; Jian WU ; Yimin XU ; Hong YAO ; Dongsheng YANG ; Xu YANG ; Yanling YANG ; Ying ZHANG ; Yulin ZHOU ; Baosheng ZHU ; Sicong ZENG ; Zhiyu PENG ; Shangzhi HUANG
Chinese Journal of Medical Genetics 2018;35(1):1-8
The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.
4.Preliminary bioinformatics analyses of the expression and function of AGR2 in human breast cancer
QIU Yiran ; LIU Wenjia ; YU Yue ; CAO Xuchen
Chinese Journal of Cancer Biotherapy 2020;27(3):302-308
Objective: To explore the expression of AGR2 gene in breast cancer as well as to predict its relevant biological functions and molecular signaling pathways with bioinformatics tool. Methods: The expression of AGR2 in breast cancer tissues and normal tissues was analyzed in Oncomine and GEPIA databases, and the expression of AGR2 in breast cancer cell lines was evaluated in CCLE database. Meanwhile, HPA database was used to analyze the expression of AGR2 protein in normal and breast cancer tissues. Besides, the gene expression microarray data download from CCLE database was analyzed by using R software to obtain genes co-expressed withAGR2. Functional annotation ofAGR2 co-expressed genes was performed by using GO Enrichment and KEGG pathway analyses. Results: Oncomine and GEPIA databases showed that AGR2 gene was highly expressed in breast cancer tissues, and CCLE database analysis showed that AGR2 was highly expressed in all breast cancer cell lines. Immunohistochemistry results from the HPA database showed that the expression of AGR2 protein was significantly higher in breast cancer tissues compared with normal tissues. A total of 946 genes co-expressed with AGR2 in breast cancer were screened out with the R software. With the GO function Enrichment analysis, the co-expressed genes were demonstrated to be mainly involved in biological functions, such as protein localization to cell periphery, protein localization to plasma membrane, cell junction assembly, cell-substrate adhesion, and cell junction organization etc. In addition, the KEGG analysis results showed that co-expressed genes were mainly involved in the progression of gastric cancer and breast cancer, and were associated with proteoglycans in cancer, as well as proline, leucine and isoleucine degradation pathways. Conclusions:AGR2 is highly expressed in breast cancer tissues, which may be a potential oncogenic gene and a new therapeutic target of breast cancer.
5.A core epitope targeting antibody of SARS-CoV-2.
Simeng ZHAO ; Fengjiang LIU ; Shizhen QIU ; Qiaoshuai LAN ; Yiran WU ; Wei XU ; Junzi KE ; Jie YANG ; Xiaoyan LIU ; Kun WANG ; Hangtian GUO ; Shuai XIA ; Fangfang ZHANG ; Jiabei WANG ; Xiaowen HU ; Lu LU ; Shibo JIANG ; Suwen ZHAO ; Lianxin LIU ; Youhua XIE ; Xiuna YANG ; Haopeng WANG ; Guisheng ZHONG
Protein & Cell 2023;14(1):74-78