1,25-dihydroxyvitamin D3 (1,25(OH)2D3), affects enteric glial cells (EGCs) activity, but the mechanism is still unknown. The current study aimed to explore whether 1,25(OH)2D3 could regulate EGCs activity via butyrate pathway in a high-fat diet model. Male C57BL/6 J mice were fed with standard diet (SDD), or vitamin-D-deficient diet (VDD), or high-fat diet (HFD), or HFD plus sodium butyrate (SBR), or HFD plus 1,25(OH)2D3, or HFD plus S100B inhibitor ONO-2506 in vivo. CRL-2690 and Caco-2 cells were treated with palmitic acid (PA) and oleic acid (OA) complex, or S100B, or S100B plus butyric acid (BA) in vitro. 25(OH)D3, 1,25(OH)2D3, TNF-α and S100B concentrations were assayed by enzyme-linked immuno- sorbent assay (ELISA). Colonic mucosal permeability was measured by using FITC-dextran 4 kDa. Colonic butyrate was detected using high-performance liquid chromatography (HPLC). The results showed HFD decreased serum 25(OH)D3 and 1,25(OH)2D3 concentrations and colonic butyrate generation. 1,25(OH)2D3 supplementation raised butyrate production in the colon. 1,25(OH)2D3 and sodium butyrate supplementation inhibited EGCs to produce S100B and reduced colonic permeability to FITC-dextran. Inhibition of S100B pathway by ONO- 2506 decreased colonic hyperpermeability. In vitro experiments showed butyrate treatment not only reduced S100B and TNF-α secretion from PA/OA-treated CRL-2690 cells, but also decreased the permeability of S100B-treated Caco-2 cells. Collectively, 1,25(OH)2D3 elicited butyrate to suppress EGCs activation, which helped to prevent intestinal barrier injury.
{"title":"1,25-dihydroxyvitamin D3 regulates enteroglial bioactivity through butyric acid pathway in a high-fat diet mouse model.","authors":"Aiwen Feng, Shaosheng Su, Qian Li, Cheng Li, Yingyan Liu, Jiasheng Qiu","doi":"10.1016/j.jsbmb.2024.106655","DOIUrl":"10.1016/j.jsbmb.2024.106655","url":null,"abstract":"<p><p>1,25-dihydroxyvitamin D3 (1,25(OH)2D3), affects enteric glial cells (EGCs) activity, but the mechanism is still unknown. The current study aimed to explore whether 1,25(OH)2D3 could regulate EGCs activity via butyrate pathway in a high-fat diet model. Male C57BL/6 J mice were fed with standard diet (SDD), or vitamin-D-deficient diet (VDD), or high-fat diet (HFD), or HFD plus sodium butyrate (SBR), or HFD plus 1,25(OH)2D3, or HFD plus S100B inhibitor ONO-2506 in vivo. CRL-2690 and Caco-2 cells were treated with palmitic acid (PA) and oleic acid (OA) complex, or S100B, or S100B plus butyric acid (BA) in vitro. 25(OH)D3, 1,25(OH)2D3, TNF-α and S100B concentrations were assayed by enzyme-linked immuno- sorbent assay (ELISA). Colonic mucosal permeability was measured by using FITC-dextran 4 kDa. Colonic butyrate was detected using high-performance liquid chromatography (HPLC). The results showed HFD decreased serum 25(OH)D3 and 1,25(OH)2D3 concentrations and colonic butyrate generation. 1,25(OH)2D3 supplementation raised butyrate production in the colon. 1,25(OH)2D3 and sodium butyrate supplementation inhibited EGCs to produce S100B and reduced colonic permeability to FITC-dextran. Inhibition of S100B pathway by ONO- 2506 decreased colonic hyperpermeability. In vitro experiments showed butyrate treatment not only reduced S100B and TNF-α secretion from PA/OA-treated CRL-2690 cells, but also decreased the permeability of S100B-treated Caco-2 cells. Collectively, 1,25(OH)2D3 elicited butyrate to suppress EGCs activation, which helped to prevent intestinal barrier injury.</p>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":" ","pages":"106655"},"PeriodicalIF":2.7,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142795666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-06DOI: 10.1016/j.jsbmb.2024.106653
Xiaomeng Pei, Haolin Li, Hao Yu, Wei Wang, Dagan Mao
AMPK plays a crucial role in cellular energy metabolism and is involved in the regulation of luteal steroidogenesis by APN and its analog AdipoRon. To further explore the regulatory mechanism of AMPK in goat luteal steroidogenesis mediated by APN, cyclic and pregnant CL were utilized to assess the localization and expression of AMPK, EZH2, H3K27me3 and H3K27ac by WB and mIHC, and the interaction between AMPK and EZH2 by Co-IP. Then, isolated luteal cells were treated with APN/AdipoRon to evaluate the expression levels of AMPK, EZH2, H3K27me3 and H3K27ac. Results showed that AMPK and EZH2 were co-localized to the cytoplasm of luteal cells, and interacted as detected by Co-IP. H3K27me3 and H3K27ac were localized to the nucleus of goat luteal cells. H3K27me3 expression in late CL was significantly higher than that in early and middle CL, while the expressions of AMPK, H3K27ac and EZH2 in middle CL were significantly higher than those in early and late CL. Notably, all these proteins were expressed at similar levels between pregnancy and middle cycle, with the exception of EZH2. Following incubation with AdipoRon (25 μM) and APN (1 μg/mL) for 24 h, the expressions of AMPK and H3K27ac decreased, while H3K27me3 increased in luteal cells. Compound C (AMPK activity inhibitor) reversed the AdipoRon - induced decrease in EZH2 expression and the increase in H3K27me3 expression. The increased H3K27me3 expression and decreased steroidogenic protein (CYP11A1 and HSD3B) expression after GSK126 (EZH2 inhibitor) treatment were consistent with the effects seen after AdipoRon treatment. In conclusion, APN/AdipoRon inhibits luteal steroidogenesis by inhibiting the interaction between AMPK and EZH2, thereby promoting H3K27me3 expression.
{"title":"APN/AdipoRon regulates luteal steroidogenesis through AMPK/EZH2/H3K27me3 in goats.","authors":"Xiaomeng Pei, Haolin Li, Hao Yu, Wei Wang, Dagan Mao","doi":"10.1016/j.jsbmb.2024.106653","DOIUrl":"10.1016/j.jsbmb.2024.106653","url":null,"abstract":"<p><p>AMPK plays a crucial role in cellular energy metabolism and is involved in the regulation of luteal steroidogenesis by APN and its analog AdipoRon. To further explore the regulatory mechanism of AMPK in goat luteal steroidogenesis mediated by APN, cyclic and pregnant CL were utilized to assess the localization and expression of AMPK, EZH2, H3K27me3 and H3K27ac by WB and mIHC, and the interaction between AMPK and EZH2 by Co-IP. Then, isolated luteal cells were treated with APN/AdipoRon to evaluate the expression levels of AMPK, EZH2, H3K27me3 and H3K27ac. Results showed that AMPK and EZH2 were co-localized to the cytoplasm of luteal cells, and interacted as detected by Co-IP. H3K27me3 and H3K27ac were localized to the nucleus of goat luteal cells. H3K27me3 expression in late CL was significantly higher than that in early and middle CL, while the expressions of AMPK, H3K27ac and EZH2 in middle CL were significantly higher than those in early and late CL. Notably, all these proteins were expressed at similar levels between pregnancy and middle cycle, with the exception of EZH2. Following incubation with AdipoRon (25 μM) and APN (1 μg/mL) for 24 h, the expressions of AMPK and H3K27ac decreased, while H3K27me3 increased in luteal cells. Compound C (AMPK activity inhibitor) reversed the AdipoRon - induced decrease in EZH2 expression and the increase in H3K27me3 expression. The increased H3K27me3 expression and decreased steroidogenic protein (CYP11A1 and HSD3B) expression after GSK126 (EZH2 inhibitor) treatment were consistent with the effects seen after AdipoRon treatment. In conclusion, APN/AdipoRon inhibits luteal steroidogenesis by inhibiting the interaction between AMPK and EZH2, thereby promoting H3K27me3 expression.</p>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":" ","pages":"106653"},"PeriodicalIF":2.7,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142795687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-05DOI: 10.1016/j.jsbmb.2024.106654
Ana Raquel Pereira da Silva, Maria do Socorro Costa, Nara Juliana Santos Araújo, Thiago Sampaio de Freitas, Cícera Laura Roque Paulo, Maria Anésia Sousa de Alencar, José Maria Barbosa-Filho, Jacqueline Cosmo Andrade-Pinheiro, Henrique Douglas Melo Coutinho
Biofilms are complex microbial structures that have a significant impact on human health, industry and the environment. These complex structures represent one of the main mechanisms of microbial resistance, and their development constitutes a serious health problem. Therefore, the aim of this study was to verify the potential for inhibition and eradication of bacterial biofilm by salosodine, which is a steroidal alkaloid sapogenin found in plants of the Solanum genus. The antibiotics gentamicin, norfloxacin, ampicillin and the antiseptic agent chlorhexidine gluconate were used as positive controls to compare the results. Solasodin showed significant results in inhibiting the formation of Enterococcus faecalis and Staphylococcus aureus biofilms at the two concentrations tested. And when comparing the effect of solasodine for the two concentrations and the effect of the antibiotic gentamicin, it was found that sapogenin showed a better percentage in inhibiting E. faecalis biofilm formation. And against Pseudomonas aeruginosa, solasodine only inhibited biofilm formation at the highest concentration compared to the control. In the biofilm eradication results, solasodine showed a significant reduction in the biomass of the S. aureus biofilm, and when compared with the percentage reduction of the antibiotics, solasodine showed a relevant result for both concentrations. Only at the lowest concentration did solasodine show a reduction in P. aeruginosa biofilm biomass, a reduction close to that of chlorhexidine gluconate. In terms of activity, solasodine has been shown to have the potential to inhibit biofilm formation. However, further tests are needed to investigate the mechanisms of action of this sapogenin on the bacterial biofilms tested.
{"title":"Evaluation of inhibition and eradication of bacterial biofilm by solasodin.","authors":"Ana Raquel Pereira da Silva, Maria do Socorro Costa, Nara Juliana Santos Araújo, Thiago Sampaio de Freitas, Cícera Laura Roque Paulo, Maria Anésia Sousa de Alencar, José Maria Barbosa-Filho, Jacqueline Cosmo Andrade-Pinheiro, Henrique Douglas Melo Coutinho","doi":"10.1016/j.jsbmb.2024.106654","DOIUrl":"10.1016/j.jsbmb.2024.106654","url":null,"abstract":"<p><p>Biofilms are complex microbial structures that have a significant impact on human health, industry and the environment. These complex structures represent one of the main mechanisms of microbial resistance, and their development constitutes a serious health problem. Therefore, the aim of this study was to verify the potential for inhibition and eradication of bacterial biofilm by salosodine, which is a steroidal alkaloid sapogenin found in plants of the Solanum genus. The antibiotics gentamicin, norfloxacin, ampicillin and the antiseptic agent chlorhexidine gluconate were used as positive controls to compare the results. Solasodin showed significant results in inhibiting the formation of Enterococcus faecalis and Staphylococcus aureus biofilms at the two concentrations tested. And when comparing the effect of solasodine for the two concentrations and the effect of the antibiotic gentamicin, it was found that sapogenin showed a better percentage in inhibiting E. faecalis biofilm formation. And against Pseudomonas aeruginosa, solasodine only inhibited biofilm formation at the highest concentration compared to the control. In the biofilm eradication results, solasodine showed a significant reduction in the biomass of the S. aureus biofilm, and when compared with the percentage reduction of the antibiotics, solasodine showed a relevant result for both concentrations. Only at the lowest concentration did solasodine show a reduction in P. aeruginosa biofilm biomass, a reduction close to that of chlorhexidine gluconate. In terms of activity, solasodine has been shown to have the potential to inhibit biofilm formation. However, further tests are needed to investigate the mechanisms of action of this sapogenin on the bacterial biofilms tested.</p>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":" ","pages":"106654"},"PeriodicalIF":2.7,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142792734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-20DOI: 10.1016/j.jsbmb.2024.106641
Andrea Andress Huacachino , Anna Chung , Kim Sharp , Trevor M. Penning
Per- and polyfluoroalkyl substances (PFAS) are ubiquitous environmental pollutants that are highly stable synthetic organofluorine compounds. One congener perfluorooctanoic acid (PFOA) can be detected in nearly all humans and is recognized as an endocrine disrupting chemical (EDC). EDCs disrupt hormone synthesis and metabolism and receptor function. One mechanism of steroid hormone action is the pre-receptor regulation of ligand access to steroid hormone receptors by aldo-keto reductases. Here we report PFOA inhibition of AKR family 1 member C2 (AKR1C2), leading to dysregulation of androgen action. Spectrofluorimetric inhibitor screens identified PFOA as a competitive and tight binding inhibitor of AKR1C2, whose role is to inactivate 5α-dihydrotestosterone (5α-DHT). Further site directed mutagenesis studies along with molecular docking simulations revealed the importance of residue Valine 54 in mediating AKR1C2 inhibitor specificity. Binding site restrictions were explored by testing inhibition of other related PFAS chemicals, confirming that steric hinderance is a key factor. Furthermore, radiochromatography using HPLC and in line radiometric detection confirmed the accumulation of 5α-DHT as a result of PFOA inhibition of AKR1C2. We showed that PFOA could enhance the transactivation of AR in reporter genes assays in which 5α-DHT metabolism was blocked by AKR1C2 inhibition in HeLa cells. Taken together, these data suggest PFOA has a role in disrupting androgen action through inhibiting AKR1C2. Our work identifies an EDC function for PFOA not previously revealed.
{"title":"Specific and potent inhibition of steroid hormone pre-receptor regulator AKR1C2 by perfluorooctanoic acid: Implications for androgen metabolism","authors":"Andrea Andress Huacachino , Anna Chung , Kim Sharp , Trevor M. Penning","doi":"10.1016/j.jsbmb.2024.106641","DOIUrl":"10.1016/j.jsbmb.2024.106641","url":null,"abstract":"<div><div>Per- and polyfluoroalkyl substances (PFAS) are ubiquitous environmental pollutants that are highly stable synthetic organofluorine compounds. One congener perfluorooctanoic acid (PFOA) can be detected in nearly all humans and is recognized as an endocrine disrupting chemical (EDC). EDCs disrupt hormone synthesis and metabolism and receptor function. One mechanism of steroid hormone action is the pre-receptor regulation of ligand access to steroid hormone receptors by aldo-keto reductases. Here we report PFOA inhibition of AKR family 1 member C2 (AKR1C2), leading to dysregulation of androgen action. Spectrofluorimetric inhibitor screens identified PFOA as a competitive and tight binding inhibitor of AKR1C2, whose role is to inactivate 5α-dihydrotestosterone (5α-DHT). Further site directed mutagenesis studies along with molecular docking simulations revealed the importance of residue Valine 54 in mediating AKR1C2 inhibitor specificity. Binding site restrictions were explored by testing inhibition of other related PFAS chemicals, confirming that steric hinderance is a key factor. Furthermore, radiochromatography using HPLC and in line radiometric detection confirmed the accumulation of 5α-DHT as a result of PFOA inhibition of AKR1C2. We showed that PFOA could enhance the transactivation of AR in reporter genes assays in which 5α-DHT metabolism was blocked by AKR1C2 inhibition in HeLa cells. Taken together, these data suggest PFOA has a role in disrupting androgen action through inhibiting AKR1C2. Our work identifies an EDC function for PFOA not previously revealed.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"246 ","pages":"Article 106641"},"PeriodicalIF":2.7,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142689428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-20DOI: 10.1016/j.jsbmb.2024.106640
Sam Kafai Yahyavi , Rune Holt , Mads Joon Jorsal , Lív Bech Árting , Ebbe Eldrup , Anders Juul , Niels Jørgensen , Martin Blomberg Jensen
Background and objective
While all types of vitamin D metabolites are bound to vitamin D binding protein and albumin leaving only a small fraction in its free active form, only serum concentrations of total 25-hydroxy vitamin D (25OHD) are used to determine vitamin D status in clinical practice. This study aimed to describe the association of total 25-hydroxy vitamin D (25OHD), calculated free 25OHD, and free 25OHD% (free 25OHD × 100 %/total 25OHD) with mineral, bone, and metabolic variables and assess the impact of cholecalciferol supplementation.
Research design and methods
Secondary data from a single-center, double-blinded, randomized, placebo-controlled clinical trial (NCT01304927) in 307 infertile men. The treatment group (n = 151) initially received 300,000 IU cholecalciferol as a bolus followed by 1400 IU daily for 150 days and was compared to a placebo group (n = 156).
Results
At baseline men with free 25OHD% > 0.03 % had lower serum triglycerides (mmol/L) (0.8 vs. 1.0; p = 0.002), lower LDL (mmol/L) (2.7 vs. 3.1; p = 0.003), lower fasting blood glucose (mmol/L) (4.9 vs. 5.2; p = 0.012), and lower PTH (pmol/L) (3.8 vs. 4.6; p = 0.015) compared to men with free 25OHD% < 0.02 %. When the study population was stratified according to total 25OHD or free 25OHD, the metabolic markers and bone variables did not show any differences. Cholecalciferol supplementation increased total 25OHD after 150 days compared to placebo and the difference was highest in men with lowest vitamin D status. Cholecalciferol supplementation did not change free 25OHD%.
Conclusion
The free 25OHD% is better associated with metabolic health markers such as serum triglycerides, LDL, and fasting blood glucose, but not bone or calciotrophic markers except parathyroid hormone. The free 25OHD% is not affected by cholecalciferol supplementation.
背景和目的：虽然所有类型的维生素 D 代谢物都与维生素 D 结合蛋白和白蛋白结合，只留下一小部分以游离活性形式存在，但在临床实践中，只有血清中总 25- 羟基维生素 D (25OHD) 的浓度被用来确定维生素 D 状态。本研究旨在描述总25-羟基维生素D（25OHD）、计算出的游离25OHD和游离25OHD%（游离25OHD x100%/总25OHD）与矿物质、骨骼和代谢变量的关系，并评估补充胆钙化醇的影响：307名不育男性的单中心、双盲、随机、安慰剂对照临床试验（NCT01304927）的二次数据。治疗组（人数=151）最初接受30万IU胆钙化醇的栓剂治疗，然后在150天内每天服用1400IU，并与安慰剂组（人数=156）进行比较：基线时游离25OHD%>0.03%的男性血清甘油三酯（mmol/L）较低（0.8 vs. 1.0；p=0.002），低密度脂蛋白（mmol/L）较低（2.7 vs. 3.1；p=0.003）、较低的空腹血糖（mmol/L）（4.9 vs. 5.2；p=0.012）和较低的 PTH（pmol/L）（3.8 vs. 4.6；p=0.015）：游离 25OHD% 与血清甘油三酯、低密度脂蛋白和空腹血糖等代谢健康指标有较好的相关性，但与除甲状旁腺激素外的骨或钙营养指标无关。游离 25OHD% 不受胆钙化醇补充剂的影响。
{"title":"Influence of cholecalciferol supplementation on changes in total 25OHD, free 25OHD, and free 25OHD % in relation to calcium, bone, and glucose homeostasis in young, infertile men","authors":"Sam Kafai Yahyavi , Rune Holt , Mads Joon Jorsal , Lív Bech Árting , Ebbe Eldrup , Anders Juul , Niels Jørgensen , Martin Blomberg Jensen","doi":"10.1016/j.jsbmb.2024.106640","DOIUrl":"10.1016/j.jsbmb.2024.106640","url":null,"abstract":"<div><h3>Background and objective</h3><div>While all types of vitamin D metabolites are bound to vitamin D binding protein and albumin leaving only a small fraction in its free active form, only serum concentrations of total 25-hydroxy vitamin D (25OHD) are used to determine vitamin D status in clinical practice. This study aimed to describe the association of total 25-hydroxy vitamin D (25OHD), calculated free 25OHD, and free 25OHD% (free 25OHD × 100 %/total 25OHD) with mineral, bone, and metabolic variables and assess the impact of cholecalciferol supplementation.</div></div><div><h3>Research design and methods</h3><div>Secondary data from a single-center, double-blinded, randomized, placebo-controlled clinical trial (NCT01304927) in 307 infertile men. The treatment group (n = 151) initially received 300,000 IU cholecalciferol as a bolus followed by 1400 IU daily for 150 days and was compared to a placebo group (n = 156).</div></div><div><h3>Results</h3><div>At baseline men with free 25OHD% > 0.03 % had lower serum triglycerides (mmol/L) (0.8 vs. 1.0; p = 0.002), lower LDL (mmol/L) (2.7 vs. 3.1; p = 0.003), lower fasting blood glucose (mmol/L) (4.9 vs. 5.2; p = 0.012), and lower PTH (pmol/L) (3.8 vs. 4.6; p = 0.015) compared to men with free 25OHD% < 0.02 %. When the study population was stratified according to total 25OHD or free 25OHD, the metabolic markers and bone variables did not show any differences. Cholecalciferol supplementation increased total 25OHD after 150 days compared to placebo and the difference was highest in men with lowest vitamin D status. Cholecalciferol supplementation did not change free 25OHD%.</div></div><div><h3>Conclusion</h3><div>The free 25OHD% is better associated with metabolic health markers such as serum triglycerides, LDL, and fasting blood glucose, but not bone or calciotrophic markers except parathyroid hormone. The free 25OHD% is not affected by cholecalciferol supplementation.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"246 ","pages":"Article 106640"},"PeriodicalIF":2.7,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142693887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-19DOI: 10.1016/j.jsbmb.2024.106639
Segovia-Mendoza Mariana , Reyes-Plata Brenda , Prado-Garcia Heriberto , Lemini Cristina , Barrera David , Ángeles-López Guadalupe
Breast cancer (BC) is the most frequent female neoplasm worldwide. Its establishment and development have been related to inflammatory cytokine expression. Steroid hormones such as estradiol (E2) can regulate proinflammatory cytokine secretion through interaction with its nuclear receptors. However, little is known regarding the activation of its membrane estrogen receptor (GPER1) and the inflammatory cytokine environment in BC. We have studied the synthesis and biological effects of molecules analogs to E2 for hormone replacement therapy (HRT), such as pentolame. Nevertheless, its interaction with GPER1 and the modulation of inflammatory cytokines in different BC types has been barely studied and deserves deeper investigation. In this research, the role of GPER1 in the proliferation and modulation of inflammatory cytokines involved in carcinogenesis and metastatic processes in different BC cell lines was assessed by binding to various compounds. To achieve this goal, the presence of GPER1 was identified in different BC cell lines. Subsequently, cell proliferation after exposure to E2, pentolame and GPER1 agonist, G1, was subsequently determined alone or in combination with the GPER1 antagonist, G15. Finally, the pro-inflammatory cytokine secretion derived from the supernatants of BC cells exposed to the previous treatments was also assessed. Interestingly, GPER1 activation or inhibition has significant effects on the cytokine regulation associated with invasion in BC. Notably, pentolame did not induce cell proliferation or increase the proinflammatory cytokine expression compared to E2 in BC cell lines. In addition, pentolame did not induce the presence of the cell adhesion molecule PECAM-1. In contrast, E2 treatment weakly induced the expression of PECAM-1 in MCF-7 and HCC1937 cells, and G1 treatment showed this effect only in MCF-7 cells. The results suggest that GPER1 might be a significant inflammatory modulator with angiogenic-related effects in BC cells. In addition, pentolame might represent an HRT alternative in patients with BC predisposition.
乳腺癌（BC）是全球最常见的女性肿瘤。它的形成和发展与炎性细胞因子的表达有关。雌二醇（E2）等类固醇激素可通过与其核受体相互作用来调节促炎细胞因子的分泌。然而，人们对其膜雌激素受体（GPER1）的激活和 BC 炎症细胞因子环境知之甚少。我们研究了用于激素替代疗法（HRT）的 E2 分子类似物（如喷托胺）的合成和生物效应。然而，我们几乎没有研究过它与 GPER1 的相互作用以及对不同类型 BC 中炎症细胞因子的调节作用，这一点值得深入研究。在这项研究中，通过与各种化合物结合，评估了 GPER1 在不同 BC 细胞系的增殖和炎症细胞因子的调控中的作用。为实现这一目标，研究人员在不同的 BC 细胞系中发现了 GPER1 的存在。随后，单独或与 GPER1 拮抗剂 G15 结合测定了暴露于 E2、戊巴醇和 GPER1 激动剂 G1 后的细胞增殖情况。最后，还评估了暴露于先前处理的 BC 细胞上清液中的促炎细胞因子分泌情况。有趣的是，GPER1 的激活或抑制对与 BC 侵袭相关的细胞因子调控有显著影响。值得注意的是，与 E2 相比，在 BC 细胞系中，五氯苯胺不会诱导细胞增殖或增加促炎细胞因子的表达。此外，戊唑醇也不会诱导细胞粘附分子 PECAM-1 的存在。相比之下，E2 处理能微弱地诱导 MCF-7 和 HCC1937 细胞中 PECAM-1 的表达，而 G1 处理仅在 MCF-7 细胞中显示出这种效应。这些结果表明，GPER1 可能是一种重要的炎症调节剂，对 BC 细胞具有血管生成相关作用。此外，对于有 BC 易感性的患者来说，五氯苯胺可能是一种激素替代疗法。
{"title":"GPER1 activation by estrogenic compounds in the inflammatory profile of breast cancer cells","authors":"Segovia-Mendoza Mariana , Reyes-Plata Brenda , Prado-Garcia Heriberto , Lemini Cristina , Barrera David , Ángeles-López Guadalupe","doi":"10.1016/j.jsbmb.2024.106639","DOIUrl":"10.1016/j.jsbmb.2024.106639","url":null,"abstract":"<div><div>Breast cancer (BC) is the most frequent female neoplasm worldwide. Its establishment and development have been related to inflammatory cytokine expression. Steroid hormones such as estradiol (E<sub>2</sub>) can regulate proinflammatory cytokine secretion through interaction with its nuclear receptors. However, little is known regarding the activation of its membrane estrogen receptor (GPER1) and the inflammatory cytokine environment in BC. We have studied the synthesis and biological effects of molecules analogs to E<sub>2</sub> for hormone replacement therapy (HRT), such as pentolame. Nevertheless, its interaction with GPER1 and the modulation of inflammatory cytokines in different BC types has been barely studied and deserves deeper investigation. In this research, the role of GPER1 in the proliferation and modulation of inflammatory cytokines involved in carcinogenesis and metastatic processes in different BC cell lines was assessed by binding to various compounds. To achieve this goal, the presence of GPER1 was identified in different BC cell lines. Subsequently, cell proliferation after exposure to E<sub>2</sub>, pentolame and GPER1 agonist, G1, was subsequently determined alone or in combination with the GPER1 antagonist, G15. Finally, the pro-inflammatory cytokine secretion derived from the supernatants of BC cells exposed to the previous treatments was also assessed. Interestingly, GPER1 activation or inhibition has significant effects on the cytokine regulation associated with invasion in BC. Notably, pentolame did not induce cell proliferation or increase the proinflammatory cytokine expression compared to E<sub>2</sub> in BC cell lines. In addition, pentolame did not induce the presence of the cell adhesion molecule PECAM-1. In contrast, E2 treatment weakly induced the expression of PECAM-1 in MCF-7 and HCC1937 cells, and G1 treatment showed this effect only in MCF-7 cells. The results suggest that GPER1 might be a significant inflammatory modulator with angiogenic-related effects in BC cells. In addition, pentolame might represent an HRT alternative in patients with BC predisposition.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"245 ","pages":"Article 106639"},"PeriodicalIF":2.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142689423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-19DOI: 10.1016/j.jsbmb.2024.106638
Jie Xiang , Mingzhu Zhong , Qian Zhang , Yang Zhu , Peipei Pan , Huitao Li , Qianjin Fei , Rongying Ou , Ren-shan Ge , Weibing Zhang
Parabens are widely used as preservatives in personal care products and are linked to potential disruptions in placental steroidogenesis. However, their exact impact remains unclear. This study aimed to explore the inhibition, mechanisms, structure-activity relationships (SAR) of parabens on human placental 3β-hydroxysteroid dehydrogenase type 1 (h3β-HSD1) and its rat counterpart, r3β-HSD4.3β-HSD activity was assayed in placental microsomes using pregnenolone as substrate and HPLC-MS/MS to measure progesterone and the effects of parabens on 3β-HSD was evaluated and SAR was performed. The research identified their inhibition against h3β-HSD1, with nonylparaben showing the highest potency (IC50, 4.17 µM), followed by phenylparaben, heptylparaben, hexylparaben, benzylparaben, butylparaben, propylparaben, and ethylparaben. The inhibition was characterized as mixed/noncompetitive. Additionally, these parabens inhibited progesterone secretion in human JAr cells at ≤100 µM. Similar trends were observed for r3β-HSD4. Docking simulations indicated that parabens interact with NAD+ and steroid-binding sites of both enzymes. A negative correlation between LogP, molecular weight, volume, and alcohol chain carbon with IC50 values highlighted the role of carbon chain length in determining inhibitory efficacy. The inhibitory potency of parabens on 3β-HSD is significantly influenced by their structural attributes, particularly the length of their carbon chains and LogP values.
{"title":"Effects of parabens on human and rat placental 3β-hydroxysteroid dehydrogenase isoforms: Structure activity relationship and docking analysis","authors":"Jie Xiang , Mingzhu Zhong , Qian Zhang , Yang Zhu , Peipei Pan , Huitao Li , Qianjin Fei , Rongying Ou , Ren-shan Ge , Weibing Zhang","doi":"10.1016/j.jsbmb.2024.106638","DOIUrl":"10.1016/j.jsbmb.2024.106638","url":null,"abstract":"<div><div>Parabens are widely used as preservatives in personal care products and are linked to potential disruptions in placental steroidogenesis. However, their exact impact remains unclear. This study aimed to explore the inhibition, mechanisms, structure-activity relationships (SAR) of parabens on human placental 3β-hydroxysteroid dehydrogenase type 1 (h3β-HSD1) and its rat counterpart, r3β-HSD4.3β-HSD activity was assayed in placental microsomes using pregnenolone as substrate and HPLC-MS/MS to measure progesterone and the effects of parabens on 3β-HSD was evaluated and SAR was performed. The research identified their inhibition against h3β-HSD1, with nonylparaben showing the highest potency (IC<sub>50</sub>, 4.17 µM), followed by phenylparaben, heptylparaben, hexylparaben, benzylparaben, butylparaben, propylparaben, and ethylparaben. The inhibition was characterized as mixed/noncompetitive. Additionally, these parabens inhibited progesterone secretion in human JAr cells at ≤100 µM. Similar trends were observed for r3β-HSD4. Docking simulations indicated that parabens interact with NAD<sup>+</sup> and steroid-binding sites of both enzymes. A negative correlation between LogP, molecular weight, volume, and alcohol chain carbon with IC<sub>50</sub> values highlighted the role of carbon chain length in determining inhibitory efficacy. The inhibitory potency of parabens on 3β-HSD is significantly influenced by their structural attributes, particularly the length of their carbon chains and LogP values.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"245 ","pages":"Article 106638"},"PeriodicalIF":2.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142683458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-16DOI: 10.1016/j.jsbmb.2024.106632
Zichang Gui , Wei Shi , Fangting Zhou , Yongqing Yan , Yuntian Li , Yang Xu
To date five members of estrogen receptors (ESRs) have been reported. They are grouped into two classes, the nuclear estrogen receptors are members of the nuclear receptor family which found at nuclear, cytoplasm and plasma membrane, and the membrane estrogen receptors, such as G protein-coupled estrogen receptor 1, ESR-X and Gq-coupled membrane estrogen receptor. The structure and function of estrogen receptors, and interaction between ESR and coregulators were reviewed. In canonical pathway ESRs can translocate to the nucleus, bind to the target gene promotor with or without estrogen responsive element and regulate transcription, mediating the genomic effects of estrogen. Coactivators and corepressors are recruited to activate or inhibit transcription by activated ESRs. Many coactivators and corepressors are recruited to activate or inhibit ESR mediated gene transcription via different mechanisms. ESRs also indirectly bind to the promoter via interaction with other transcription factors, tethering the transcription factors. ESRs can be phosphorylated by several kinases such as p38, extracellular-signal-regulated kinase, and activated protein kinase B, and which activates transcription without ligand binding. Non-genomic estrogen action can be manifested by the increases of cytoplasmic NO and Ca2+ through the activation of membrane ESRs. In female, ESRs signaling is crucial for folliculogenesis, oocyte growth, ovulation, oviduct and uterus. In male, ESRs signaling modulates libido, erectile function, leydig cell steroidogenesis, sertoli cell’s function, and epididymal fluid homeostatsis, supporting spermatogenesis and sperm maturation. The abnormal ESRs signaling is believed to be closely related to reproductive diseases and cancer.
{"title":"The role of estrogen receptors in intracellular estrogen signaling pathways, an overview","authors":"Zichang Gui , Wei Shi , Fangting Zhou , Yongqing Yan , Yuntian Li , Yang Xu","doi":"10.1016/j.jsbmb.2024.106632","DOIUrl":"10.1016/j.jsbmb.2024.106632","url":null,"abstract":"<div><div>To date five members of estrogen receptors (ESRs) have been reported. They are grouped into two classes, the nuclear estrogen receptors are members of the nuclear receptor family which found at nuclear, cytoplasm and plasma membrane, and the membrane estrogen receptors, such as G protein-coupled estrogen receptor 1, ESR-X and Gq-coupled membrane estrogen receptor. The structure and function of estrogen receptors, and interaction between ESR and coregulators were reviewed. In canonical pathway ESRs can translocate to the nucleus, bind to the target gene promotor with or without estrogen responsive element and regulate transcription, mediating the genomic effects of estrogen. Coactivators and corepressors are recruited to activate or inhibit transcription by activated ESRs. Many coactivators and corepressors are recruited to activate or inhibit ESR mediated gene transcription via different mechanisms. ESRs also indirectly bind to the promoter via interaction with other transcription factors, tethering the transcription factors. ESRs can be phosphorylated by several kinases such as p38, extracellular-signal-regulated kinase, and activated protein kinase B, and which activates transcription without ligand binding. Non-genomic estrogen action can be manifested by the increases of cytoplasmic NO and Ca<sup>2+</sup> through the activation of membrane ESRs. In female, ESRs signaling is crucial for folliculogenesis, oocyte growth, ovulation, oviduct and uterus. In male, ESRs signaling modulates libido, erectile function, leydig cell steroidogenesis, sertoli cell’s function, and epididymal fluid homeostatsis, supporting spermatogenesis and sperm maturation. The abnormal ESRs signaling is believed to be closely related to reproductive diseases and cancer.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"245 ","pages":"Article 106632"},"PeriodicalIF":2.7,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142649028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-16DOI: 10.1016/j.jsbmb.2024.106634
Yuanxin Liu , Jie Qin , Xiaorui Li , Guangzhen Wu
Oxysterols are compounds generated through oxidative reactions involving cholesterol and other steroid molecules. They play a crucial role in the tumor immune microenvironment by interacting with molecules such as the cell membrane receptor EBI2 and nuclear receptors like LXR and PXR. This interaction regulates immune cell signaling pathways, affecting proliferation, apoptosis, migration, and invasion in tumor-related processes. Activating these receptors alters the function and behavior of immune cells—such as macrophages, T cells, and dendritic cells—within the tumor microenvironment, thus promoting or inhibiting tumor development. Certain oxidized steroids can increase both the number and activation of infiltrating T cells, synergizing with anti-PD-1 to enhance anti-tumor efficacy. An in-depth study of the biological mechanisms of oxidized sterols will not only enhance our understanding of the complexity of the tumor immune microenvironment but may also reveal new therapeutic targets, providing innovative strategies for tumor immunotherapy.
{"title":"Oxysterols in tumor immune microenvironment (TIME)","authors":"Yuanxin Liu , Jie Qin , Xiaorui Li , Guangzhen Wu","doi":"10.1016/j.jsbmb.2024.106634","DOIUrl":"10.1016/j.jsbmb.2024.106634","url":null,"abstract":"<div><div>Oxysterols are compounds generated through oxidative reactions involving cholesterol and other steroid molecules. They play a crucial role in the tumor immune microenvironment by interacting with molecules such as the cell membrane receptor EBI2 and nuclear receptors like LXR and PXR. This interaction regulates immune cell signaling pathways, affecting proliferation, apoptosis, migration, and invasion in tumor-related processes. Activating these receptors alters the function and behavior of immune cells—such as macrophages, T cells, and dendritic cells—within the tumor microenvironment, thus promoting or inhibiting tumor development. Certain oxidized steroids can increase both the number and activation of infiltrating T cells, synergizing with anti-PD-1 to enhance anti-tumor efficacy. An in-depth study of the biological mechanisms of oxidized sterols will not only enhance our understanding of the complexity of the tumor immune microenvironment but may also reveal new therapeutic targets, providing innovative strategies for tumor immunotherapy.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"245 ","pages":"Article 106634"},"PeriodicalIF":2.7,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142648886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-14DOI: 10.1016/j.jsbmb.2024.106633
Kerry S. Jones , Sarah R. Meadows , Georgia Billing , Albert Koulman , Ann Prentice
Vitamin D is required for healthy growth and development, but data on human milk vitamin D content is limited. We describe a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of vitamin D metabolites in human milk, and its application in samples collected on two consecutive days from women in rural Gambia. Vitamin D compounds were extracted from 1 mL of milk by liquid-liquid extraction and derivatised with 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) prior to analysis by LC-MS/MS. The limit of quantification was 0.05 nmol/L for vitamin D2, 0.025 nmol/L for vitamin D3 and 0.1 nmol/L for 25(OH)D2 and 25(OH)D3. Within- and between-day imprecision was <12 % for all analytes except vitamin D2 (14 %).
From all data combined, geometric mean (-/+ 1 SD) vitamin D3 concentration was 0.94 (0.43, 1.80) nmol/L and for 25(OH)D3 0.32 (0.23, 0.42) nmol/L. The within-person (intra-individual) coefficient of variation (%CV) was 32 % and 12 % for vitamin D3 and 25(OH)D3, respectively. Between-person (inter-individual) %CVs were 89 % and 34 % for vitamin D3 and 25(OH)D3, respectively. There was no significant association between vitamin D metabolite concentrations and milk fat (creamatocrit). Mean vitamin D content of human milk as ARA averaged 42 IU/L with 25(OH)D3 responsible for around two-thirds of the biological activity. In conclusion, this work describes a reliable LC-MS/MS method for quantification of vitamin D and 25(OH)D in low volumes of human milk providing a platform for future work. This study contributes to current understanding of variability of milk vitamin D content.
健康的生长发育需要维生素 D，但有关母乳中维生素 D 含量的数据却很有限。我们介绍了一种用于分析母乳中维生素 D 代谢物的液相色谱串联质谱（LC-MS/MS）方法，以及该方法在冈比亚农村妇女连续两天采集的样本中的应用。用液液萃取法从 1 毫升牛奶中提取维生素 D 复合物，并用 4-苯基-1,2,4-三唑啉-3,5-二酮（PTAD）衍生，然后用 LC-MS/MS 进行分析。维生素 D2 的定量限为 0.05 nmol/L，维生素 D3 为 0.025 nmol/L，25(OH)D2 和 25(OH)D3 为 0.1 nmol/L。日内和日间不精确度为 2（14%）。综合所有数据，维生素 D3 浓度的几何平均数（-/+ 1SD）为 0.94 (0.43, 1.80) nmol/L，25(OH)D3 为 0.32 (0.23, 0.42) nmol/L。维生素 D3 和 25(OH)D3 的个体内变异系数 (%CV) 分别为 32% 和 12%。维生素 D3 和 25(OH)D3 的个体间变异系数分别为 89% 和 34%。维生素 D 代谢物浓度与乳脂（乳脂率）之间无明显关联。人乳中作为 ARA 的维生素 D 平均含量为 42 IU/L，其中 25(OH)D3 约占生物活性的三分之二。总之，这项工作描述了一种可靠的 LC-MS/MS 方法，用于定量检测低浓度母乳中的维生素 D 和 25(OH)D，为今后的工作提供了一个平台。这项研究有助于人们了解牛奶中维生素 D 含量的变化。
{"title":"The validation of an LC-MS/MS method for the quantification of vitamin D metabolites in human milk and their biological variability in Gambian women","authors":"Kerry S. Jones , Sarah R. Meadows , Georgia Billing , Albert Koulman , Ann Prentice","doi":"10.1016/j.jsbmb.2024.106633","DOIUrl":"10.1016/j.jsbmb.2024.106633","url":null,"abstract":"<div><div>Vitamin D is required for healthy growth and development, but data on human milk vitamin D content is limited. We describe a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of vitamin D metabolites in human milk, and its application in samples collected on two consecutive days from women in rural Gambia. Vitamin D compounds were extracted from 1 mL of milk by liquid-liquid extraction and derivatised with 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) prior to analysis by LC-MS/MS. The limit of quantification was 0.05 nmol/L for vitamin D<sub>2</sub>, 0.025 nmol/L for vitamin D<sub>3</sub> and 0.1 nmol/L for 25(OH)D<sub>2</sub> and 25(OH)D<sub>3</sub>. Within- and between-day imprecision was <12 % for all analytes except vitamin D<sub>2</sub> (14 %).</div><div>From all data combined, geometric mean (-/+ 1 SD) vitamin D<sub>3</sub> concentration was 0.94 (0.43, 1.80) nmol/L and for 25(OH)D<sub>3</sub> 0.32 (0.23, 0.42) nmol/L. The within-person (intra-individual) coefficient of variation (%CV) was 32 % and 12 % for vitamin D<sub>3</sub> and 25(OH)D<sub>3</sub>, respectively. Between-person (inter-individual) %CVs were 89 % and 34 % for vitamin D<sub>3</sub> and 25(OH)D<sub>3</sub>, respectively. There was no significant association between vitamin D metabolite concentrations and milk fat (creamatocrit). Mean vitamin D content of human milk as ARA averaged 42 IU/L with 25(OH)D<sub>3</sub> responsible for around two-thirds of the biological activity. In conclusion, this work describes a reliable LC-MS/MS method for quantification of vitamin D and 25(OH)D in low volumes of human milk providing a platform for future work. This study contributes to current understanding of variability of milk vitamin D content.</div></div>","PeriodicalId":51106,"journal":{"name":"Journal of Steroid Biochemistry and Molecular Biology","volume":"245 ","pages":"Article 106633"},"PeriodicalIF":2.7,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142640276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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