1.Effect of influenza virus on chemokine pathways in mice lungs and intervention of Shufeng Xuanfei Jiedu formula
Qi LIU ; Jianguo WANG ; Baozhen LI ; Lu JIA ; Yu GUO ; Wanfang YANG
Chinese Critical Care Medicine 2021;33(4):405-409
Objective:To screen the differentially expressed genes on whole expression profiles of the inflammation-related cytokines in mice infected with influenza virus by the gene chip technology, and to explore the intervention effect of Shufeng Xuanfei Jiedu formula.Methods:Male ICR mice were divided into normal group (N group), influenza virus infective model group (M group), Oseltamivir control group (C group) and Shufeng Xuanfei Jiedu formula high, medium and low dose groups (SH, SM, SL groups) according to the random number table method, with 10 rats in each group. A mouse model of influenza virus pneumonia was reproduced by nasal drip of influenza virus strain FM1 (0.05 mL). In N group, 0.05 mL normal saline was used. In SH, SM and SL groups, Shufeng Xuanfei Jiedu formula was used 2 hours after intranasal infection (2 times, equal and 1/2 of the clinical treatment dose, approximately 3.8, 1.9 and 1.0 g·mL -1·d -1) for 4 days. In C group, the dosage of Oseltamivir was 2.5 g·mL -1·d -1. In N group and M group, distilled water was given (0.2 mL once a day). On the 5th day, the whole lung of mice was taken. The lung index was calculated, and the pathological sections were observed. The total RNA of lung tissue was extracted and detected after hybridization with mice whole gene expression spectrum chip to select differentially expressed genes of chemokine pathways. The expression intensity ratio of the chip probe signal in each group vs. M group was calculated, and P < 0.05 and log 2ratio > 1 were up-regulated genes, while P < 0.05 and log 2ratio < -1 were down-regulated genes. Results:Compared with the N group, the lung index in the M group was significantly higher, and pathological changes were found in lung tissue, which suggested that the model of influenza virus infection was successfully established. Compared with the M group, the lung index of mice in C, SH, SM, SL groups was significantly lower (0.96±0.14, 1.45±0.22, 1.14±0.18, 1.22±0.21 vs. 1.72±0.15, all P < 0.05), and the extent and degree of lesions were reduced, however, there was no significant difference among the groups. Gene chip analysis showed that there were more differentially expressed genes in N group vs. M group, SH group vs. M group, SM group vs. M group, SL group vs. M group. It could be used for further signal transduction pathway screening. Compared with N group, the differential gene expression of chemokine C-C ligands (CCL-3, CCL-5) and chemokine C-X-C ligands (CXCL-9, CXCL-10) in M group were significantly up-regulated [log 2 (M group/N group) were 6.64, 3.51, 5.40, 6.64, respectively]. Compared with M group, the gene expressions of CCL-3, CCL-5, CXCL-9 and CXCL-10 were significantly down-regulated in C, SH, SM and SL groups [log 2 (C group/M group) were -3.96, -2.26, -3.12, -2.40; log 2 (SH group/M group) were -5.57, -2.37, -1.57, -1.01; log 2 (SM group/M group) were -4.35, -1.47, -1.26, -1.74; log 2 (SL group/M group) were -2.86, -1.86, -1.23, -1.39, respectively]. Conclusion:Shufeng Xuanfei Jiedu formula inhibits inflammatory damage in mice after influenza virus infection by down-regulating the expressions of CCL-3, CCL-5, CXCL-9 and CXCL-10 on chemokine pathways.
2.Study of the toxicity of 1-Bromo-3-chloro-5,5-dimethylhydantoin to zebrafish.
WanFang LI ; JinFeng WEI ; HongTao JIN ; MingFang HUANG ; JingXuan ZHANG ; ChengHe LI ; ChaoJie CHEN ; Chang LIU ; AiPing WANG
Biomedical and Environmental Sciences 2011;24(4):383-390
OBJECTIVE1-Bromo-3-chloro-5,5-dimethylhydantoin (BCDMH) is a solid oxidizing biocide for water disinfection. The objective of this study was to investigate the toxic effect of BCDMH on zebrafish.
METHODSThe developmental toxicity of BCDMH on zebrafish embryos and the dose-effect relationship was determined. The effect of BCDMH exposure on histopathology and tissue antioxidant activity of adult zebrafish were observed over time.
RESULTSExposure to 4 mg/L BCDMH post-fertilization was sufficient to induce a number of developmental malformations, such as edema, axial malformations, and reductions in heart rate and hatching rate. The no observable effects concentration of BCDMH on zebrafish embryo was 0.5 mg/L. After 96 h exposure, the 50% lethal concentration (95% confidence interval (CI)) of BCDMH on zebrafish embryo was 8.10 mg/L (6.15-11.16 mg/L). The 50% inhibitory concentration (95% CI) of BCDMH on hatching rate was 7.37 mg/L (6.33-8.35 mg/L). Histopathology showed two types of responses induced by BCDMH, defensive and compensatory. The extreme responses were marked hyperplasia of the gill epithelium with lamellar fusion and epidermal peeling. The histopathologic changes in the gills after 10 days exposure were accompanied by significantly higher catalase activity and lipid peroxidation.
CONCLUSIONThese results have important implications for studies on the toxicity and use of BCDMH and its analogs.
Animals ; Antioxidants ; metabolism ; Disinfectants ; toxicity ; Dose-Response Relationship, Drug ; Embryo, Nonmammalian ; drug effects ; Hydantoins ; toxicity ; Time Factors ; Water ; chemistry ; Water Pollutants, Chemical ; toxicity ; Zebrafish
3.Study on a novel Rho kinase inhibitor WAR5 for treating EAE
Zhibin DING ; Hui ZHANG ; Xingwang YANG ; Haifei ZHANG ; Jiezhong YU ; Yanhua LI ; Chunyun LIU ; Wanfang YANG ; Junlian LI ; Qianjin FENG ; Yongfei ZHAO ; Baoguo XIAO ; Cungen MA
Chinese Journal of Pathophysiology 2014;(9):1610-1615
AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor WAR 5 on the experimental autoimmune encephalomyelitis (EAE) and its possible mechanism.METHODS: Female C57BL/6 mice were randomly divided into EAE group and WAR5 group.EAE model was induced by the application of MOG 35-55 peptide.WAR5 was in-jected intraperitoneally every other day from post-immunization (PI) day 3 to PI day 27.The clinical score and body weight were recorded every other day .On PI day 28, the animals were sacrificed and spinal cords were obtained for HE and mye-lin staining .The splenocytes were isolated and the expression of CD 16/32 and CD206 were analyzed by flow cytometry . The protein extracts from the brains and spinal cords were collected for the measurement of inducible nitric oxide synthase ( iNOS) by Western blotting .RESULTS:The administration of WAR 5 delayed the onset of EAE and attenuated the clini-cal symptoms .The results of the pathological examination revealed that WAR 5 inhibited the infiltration of inflammatory cells and improved myelination in spinal cords , accompanied with the poralization of M 1 macrophages to M2 phenotype in the spleen.WAR5 inhibited the expression of iNOS in the central nervous system , especially in the spinal cords .CON-CLUSION:The therapeutic effect of WAR5 on EAE may be related to the shift of M1 macrophages to M2 phenotype and inhibition of inflammation in the central nerve system .
4.Effect of microRNA-181b-5p on the proliferation and invasion of cutaneous melanoma cells and its mechanisms
Li XIA ; Linhong YANG ; Li XU ; Wenguo SUN ; Liang YU ; Wanfang ZHAI ; Dongxia WANG ; Xiaowan KUANG
Chinese Journal of Dermatology 2022;55(7):588-595
Objective:To explore whether microRNA (miRNA) -181b-5p inhibits the proliferation and invasion of cutaneous melanoma cells by targeting pleckstrin (PLEK) .Methods:Bioinformatics methods were used to analyze cutaneous melanoma-associated core genes; dual-luciferase reporter assay was performed to verify the targeted interaction between miRNA-181b-5p and PLEK. Oligo RNA and small interfering RNA (siRNA) were used to regulate the expression of miRNA-181b-5p and PLEK in A375 cells respectively in this experiment, and A375 cells were divided into the following groups in detail: mimic negative control group, miRNA-181b-5p mimic group, inhibitor negative control group, miRNA-181b-5p inhibitor group, PLEK siRNA group, siRNA negative control group, miRNA-181b-5p inhibitor + control siRNA co-transfection group and miRNA-181b-5p inhibitor + PLEK siRNA3 co-transfection group. After 48-hour treatment, qPCR was performed to determine the mRNA expression of miRNA-181b-5p and PLEK in A375 cells, Western blot analysis to determine the PLEK protein expression, and Transwell assay to assess the invasive ability of A375 cells; after additional 24-96 hours of culture, cell counting kit-8 (CCK8) assay was conducted to assess the proliferative ability of A375 cells.Results:PLEK was the core gene for cutaneous melanoma. PLEK expression in the cutaneous melanoma in situ tissues was significantly higher than that in the paracancerous tissues ( P = 0.031) , but lower than that in the metastatic tissues ( P = 0.001) . Compared with human epidermal melanocytes HEMa-LP, the mRNA and protein expression of PLEK significantly increased in A375 cells (mRNA: 3.884 ± 0.156 vs. 0.997 ± 0.010, t = 18.48, P < 0.001; protein: 2.840 ± 0.301 vs. 1.029 ± 0.094, t = 5.47, P = 0.005) , but the miRNA-181b-5p expression significantly decreased in A375 cells (0.333 ± 0.042 vs. 0.967 ± 0.069, t = 7.83, P = 0.001) . Dual-luciferase reporter assay showed targeted binding of miRNA-181b-5p to PLEK. Compared with the mimic negative control group, the miRNA-181b-5p mimic group showed significantly decreased survival rate of A375 cells (48 hours: t = 7.96, P = 0.015; 72 hours: t = 7.50, P = 0.002; 96 hours: t = 7.96, P = 0.001) , and significantly decreased invasive ability of A375 cells ( t = 5.07, P = 0.007) ; on the contrary, the survival rate and invasive ability of A375 cells were significantly higher in the miRNA-181b-5p inhibitor group than in the inhibitor negative control group (survival rate: 24 hours, t =5.38, P = 0.013; 48 hours, t = 5.36, P = 0.013; 72 hours, t =7.63, P = 0.005; 96 hours, t = 5.99, P = 0.004; invasive ability: t = 7.24, P = 0.002) ; compared with the siRNA negative control group, the proliferative and invasive ability of A375 cells significantly decreased in the PLEK siRNA group (proliferative ability: 48, 72, 96 hours, P = 0.015, 0.011, 0.001, respectively; invasive ability: t = 4.93, P = 0.008) ; compared with the miRNA-181b-5p inhibitor + control siRNA co-transfection group, the miRNA-181b-5p inhibitor + PLEK siRNA co-transfection group showed significantly decreased proliferation rate and invasive ability of A375 cells (proliferation rate: 24, 48, 72, 96 hours, P = 0.042, 0.042, 0.037, 0.017, respectively; invasive ability: t = 8.52, P = 0.001) . Conclusion:miRNA-181b-5p can inhibit the proliferation and invasion of cutaneous melanoma A375 cells, likely by down-regulating the PLEK expression.
5.Analysis of articles published in Chinese Journal of Pediatrics from 2005 to 2014.
Yiqian SUN ; Xin CHEN ; Wei LI ; Lan JIANG ; Hua BAI
Chinese Journal of Pediatrics 2015;53(7):485-491
OBJECTIVETo explore the status of articles published in Chinese Journal of Pediatrics from 2005 to 2014.
METHODAll the articles published in Chinese Journal of Pediatrics from 2005 to 2014 were searched at Wanfang Medical Online database. The total number and citations of articles, authors, agency, single article citation, internet downloads, columns, fund and Mesh were analyzed. The end of searching period was January 2015.
RESULTFrom 2005 to 2014, 2814 articles were published in Chinese Journal of Pediatrics, 235 to 380 articles per year. A total of 1 596 articles were cited, the citation rate was 56.16%, total number of citation was 15 428. Among single article citations, of the top 20 articles, 55% (11/20) were those published in the Standard/Protocol/Guide column. Of the top 20 papers most frequently downloaded on internet, 100% were articles published in the Standard/Protocol/Guide column. During the recent 10 years, the source of the papers published in the journal covered 31 provinces, municipalities and autonomous regions. The column that published the largest number of articles was Original Article (911, 32.05%), followed by Case Report (336,11.82%) and Review (245, 8.62%). Of the total number of articles published in the journal, 747 were supported by fund, which accounted for 26%. The articles supported by national fund accounted for 8%.
CONCLUSIONArticles published in Chinese Journal of Pediatrics had high-quality and can reflect the development and research progress in pediatric medicine. It is one of the most important information resources in pediatric academic fields in China.
Bibliometrics ; China ; Pediatrics ; Periodicals as Topic ; statistics & numerical data
6.Integrated mass spectrometry imaging reveals spatial-metabolic alteration in diabetic cardiomyopathy and the intervention effects of ferulic acid
Yanhua LIU ; Xin ZHANG ; Shu YANG ; Zhi ZHOU ; Lu TIAN ; Wanfang LI ; Jinfeng WEI ; Zeper ABLIZ ; Zhonghua WANG
Journal of Pharmaceutical Analysis 2023;13(12):1496-1509
Diabetic cardiomyopathy(DCM)is a metabolic disease and a leading cause of heart failure among people with diabetes.Mass spectrometry imaging(MSI)is a versatile technique capable of combining the molecular specificity of mass spectrometry(MS)with the spatial information of imaging.In this study,we used MSI to visualize metabolites in the rat heart with high spatial resolution and sensitivity.We optimized the air flow-assisted desorption electrospray ionization(AFADESI)-MSI platform to detect a wide range of metabolites,and then used matrix-assisted laser desorption ionization(MALDI)-MSI for increasing metabolic coverage and improving localization resolution.AFADESI-MSI detected 214 and 149 metabolites in positive and negative analyses of rat heart sections,respectively,while MALDI-MSI detected 61 metabolites in negative analysis.Our study revealed the heterogenous metabolic profile of the heart in a DCM model,with over 105 region-specific changes in the levels of a wide range of metabolite classes,including carbohydrates,amino acids,nucleotides,and their derivatives,fatty acids,glycerol phospholipids,carnitines,and metal ions.The repeated oral administration of ferulic acid during 20 weeks significantly improved most of the metabolic disorders in the DCM model.Our findings provide novel insights into the molecular mechanisms underlying DCM and the potential of ferulic acid as a therapeutic agent for treating this condition.
7.The lncSIL molecule exerts a negative regulatory effect on the alveolar epithelial-mesenchymal transition induced by TGF-β1 through modulation of the EZH2/P21/CDK6 signaling pathway
Wanfang Zhang ; Lin Wang ; Pengtao Pan ; Wenxin Li ; Ruili Kang ; Ziren Zhu ; Haoqin Chen ; Xinyu Fang ; Xingcan Zhang ; Yuxin Zhang ; Yiwen Jiang ; Xinyan Li ; Benqi Yuan
Acta Universitatis Medicinalis Anhui 2024;59(4):600-604
Objective :
To investigate the role of lncSIL in transforming growth factor-β1(TGF-β1)-induced alveo- lar epithelial interstitial transformation (EMT) and its related signaling pathways .
Methods :
Western blot was used to detect the effect of lncSIL silencing on the expression of E-cadherin ( E-cad) , alpha-smooth muscle actin ( α- SMA) and Collagen I (Col I) in the process of EMT induced by TGF-β1 . LncSIL interacting proteins were ana- lyzed by RNA pulldown . Western blot was used to detect the effect of overexpression or silencing of lncSIL on the expression of its target gene enhancer of zeste homolog 2 (EZH2) and its downstream factors P21 and cyclin-de- pendent kinase 6 (CDK6) . Flow cytometry was used to analyze the effect of lncSIL on cell cycle progression .
Results:
After lncSIL silencing , the expression of α-SMA and Col I increased , the expression of E-cad decreased . RNA pulldown assay showed that EZH2 was the target protein that interacted with lncSIL , and the expression of EZH2 increased after silencing lncSIL , the expression of EZH2 downstream gene P21 decreased , CDK6 increased . Flow cytometry showed that the number of cells in S phase significantly increased . When lncSIL was overexpressed , the expression of EZH2 and CDK6 was down-regulated , the expression of P21 was up-regulated , and the number of S phase cells significantly decreased .
Conclusion
LncSIL inhibits TGF-β1-induced alveolar epithelial cell mesen- chymal transition by negatively regulating EZH2/P21 /CDK6 signaling pathway to inhibit cell cycle progression .
8.Epidemiological characteristics of local outbreak of COVID-19 caused by SARS-CoV-2 Delta variant in Liwan district, Guangzhou.
WenYan LI ; ZhiCheng DU ; Ying WANG ; Xiao LIN ; Long LU ; Qiang FANG ; WanFang ZHANG ; MingWei CAI ; Lin XU ; YuanTao HAO
Chinese Journal of Epidemiology 2021;42(10):1763-1768
9.Spatial-resolved metabolomics reveals tissue-specific metabolic reprogramming in diabetic nephropathy by using mass spectrometry imaging.
Zhonghua WANG ; Wenqing FU ; Meiling HUO ; Bingshu HE ; Yaqi LIU ; Lu TIAN ; Wanfang LI ; Zhi ZHOU ; Baili WANG ; Jianzhen XIA ; Yanhua CHEN ; Jinfeng WEI ; Zeper ABLIZ
Acta Pharmaceutica Sinica B 2021;11(11):3665-3677
Detailed knowledge on tissue-specific metabolic reprogramming in diabetic nephropathy (DN) is vital for more accurate understanding the molecular pathological signature and developing novel therapeutic strategies. In the present study, a spatial-resolved metabolomics approach based on air flow-assisted desorption electrospray ionization (AFADESI) and matrix-assisted laser desorption ionization (MALDI) integrated mass spectrometry imaging (MSI) was proposed to investigate tissue-specific metabolic alterations in the kidneys of high-fat diet-fed and streptozotocin (STZ)-treated DN rats and the therapeutic effect of astragaloside IV, a potential anti-diabetic drug, against DN. As a result, a wide range of functional metabolites including sugars, amino acids, nucleotides and their derivatives, fatty acids, phospholipids, sphingolipids, glycerides, carnitine and its derivatives, vitamins, peptides, and metal ions associated with DN were identified and their unique distribution patterns in the rat kidney were visualized with high chemical specificity and high spatial resolution. These region-specific metabolic disturbances were ameliorated by repeated oral administration of astragaloside IV (100 mg/kg) for 12 weeks. This study provided more comprehensive and detailed information about the tissue-specific metabolic reprogramming and molecular pathological signature in the kidney of diabetic rats. These findings highlighted the promising potential of AFADESI and MALDI integrated MSI based metabolomics approach for application in metabolic kidney diseases.