Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the pr...Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the primary visual cortex(V1)is altered in glaucoma.This study used DBA/2J mice as a model for spontaneous secondary glaucoma.The aim of the study was to compare the electrophysiological and histomorphological chara cteristics of neurons in the V1between 9-month-old DBA/2J mice and age-matched C57BL/6J mice.We conducted single-unit recordings in the V1 of light-anesthetized mice to measure the visually induced responses,including single-unit spiking and gamma band oscillations.The morphology of layerⅡ/Ⅲneurons was determined by neuronal nuclear antigen staining and Nissl staining of brain tissue sections.Eighty-seven neurons from eight DBA/2J mice and eighty-one neurons from eight C57BL/6J mice were examined.Compared with the C57BL/6J group,V1 neurons in the DBA/2J group exhibited weaker visual tuning and impaired spatial summation.Moreove r,fewer neuro ns were observed in the V1 of DBA/2J mice compared with C57BL/6J mice.These findings suggest that DBA/2J mice have fewer neurons in the VI compared with C57BL/6J mice,and that these neurons have impaired visual tuning.Our findings provide a better understanding of the pathological changes that occur in V1 neuron function and morphology in the DBA/2J mouse model.This study might offer some innovative perspectives regarding the treatment of glaucoma.展开更多
Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, trad...Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, traditional Chinese medicine has become a feasible alternative for certain diseases, including recurrent spontaneous abortion. However, because of the complex composition of the traditional Chinese medicine formula, its action mechanism remains unclear. Methods: We selected two isolated active ingredients (RAMP and baicalin) from the traditional Chinese medicine formula and used an abortion-prone CBA/J × DBA/2 model to simulate human RSA and compared the changes in fetal resorption rate, Treg cell percentage, and relevant cytokines before and after combination therapy. In addition, The mechanisms were preliminarily discussed using in vitro differentiation models. Results: In CBA/J × DBA/2 abortion-prone mice, the combination therapy resulted in a lower embryo resorption rate compared to that obtained with individual delivery of either RAMP or baicalin, thereby playing an embryo-protective role through the increase in Treg cells for the maintenance of maternal-fetal immune tolerance. In in vitro primary cell differentiation experiments, the concentration of Treg cells significantly increased from 11% to 17.9% after the combination therapy compared to that of the single administration group. Conclusion: the synergistic effects of RAMP and baicalin were responsible for Treg differentiation. The present study provides a solid basis for improving the applicability of traditional Chinese herbs in the treatment of recurrent spontaneous abortion.展开更多
背景研究表明青光眼的眼压增高与转化生长因子-β（TGF-β）促进小梁网细胞外基质的堆积以及黏附分子CD44导致房水排出阻力增加有关。传统中药青光安颗粒剂是中医临床上常用的降跟压药物，其是否通过小梁网通路发挥作用尚不清楚。目的...背景研究表明青光眼的眼压增高与转化生长因子-β（TGF-β）促进小梁网细胞外基质的堆积以及黏附分子CD44导致房水排出阻力增加有关。传统中药青光安颗粒剂是中医临床上常用的降跟压药物，其是否通过小梁网通路发挥作用尚不清楚。目的研究青光安颗粒剂对自发性青光眼模型DBA／2J小鼠房水动力学的影响及其作用机制。方法选取lO只眼压正常的3月龄雌性DBA／2J小鼠作为对照组，采用计算机随机数字分配法将20只9月龄自发性眼压升高的DBA／2J小鼠随机分为高眼压组和青光安组，青光安组小鼠采用2．5g／kg复方青光安颗粒剂灌胃，每天2次，连续15d，对照组和高眼压组小鼠以相同剂量生理盐水灌胃。采用经前房注入／抽吸系统进行眼压测量，分别以2．5、5．0μl／min的速率继续灌注，计算房水流畅系数（c）值和房水流出阻力（R）值。将3月龄DBA／2J小鼠60只按计算机随机数字分配法分为高剂量青光安组、中剂量青光安组、低剂量青光安组和空白对照组，每组各15只，分别用25．00、12．50和6．25g／kg青光安颗粒剂灌胃，空白对照组用等容量生理盐水灌胃，给药后7d麻醉条件下提取各组小鼠含青光安药物血清和空白对照动物血清，然后收集各组小鼠含小梁网巩膜组织进行培养，采用纤连蛋白（FN）、层黏连蛋白（LN）和神经元特异性烯醇化酶（NSE）免疫组织化学染色鉴定小梁网细胞。终质量浓度为0、5、10、20、50和100ng／ml的TGF-β处理小梁网细胞24h，采用MTT比色法检测各组细胞活性；用不同质量浓度含药血清培养经20ng／mlTGF-β处理的小梁细胞，分别于培养后24、48和72h采用ELISA法检测细胞上清液中TGF-β2受体质量浓度，采用Westernblot法检测各组小梁网细胞中CD44蛋白的相对表达量。结果对照组、高眼压组和青光安组小鼠在2．5td／min和5．0μl／min灌注速率下眼压明显不同，高眼压组小鼠眼压值明显高于青光安组和对照组，青光安组小鼠c值较高眼压组明显降低（1．08±O．36％2．35±1．34），R值明显增加（1．05±O．47 vs 0．64±0．55），差异均有统计学意义（均P〈O．01）。原代培养的细胞呈长梭形，FN、LN和NSE均呈阳性表达。5、10和20ng／mlTGF．p处理组细胞活力值明显低于0ng／mlTGF-β处理组细胞，差异均有统计学意义（均P〈0．05）。与空白对照组比较，TGF．母组小梁网细胞上清液中TGF-β2受体质量浓度和细胞中CD44蛋白相对表达量均明显升高，差异均有统计学意义（均P〈O．01）；细胞培养后24、48和72hTGF-β高剂量含药血清组细胞上清液中TGF-β2受体质量浓度和细胞中CD44相对表达量均明显低于TGF-β组和TGF-B＋低剂量含药血清组，差异均有统计学意义（均P〈0．01）。结论青光安颗粒剂通过影响房水动力学因素达到降低青光眼眼压的作用。青光安含药血清能明显降低体外培养的鼠小梁网细胞中TGF-β2受体和CD44的表达。展开更多
基金supported by the STI 2030-Major Projects 2022ZD0208500(to DY)the National Natural Science Foundation of China,Nos.82072011(to YX),82121003(to DY),82271120(to YS)+2 种基金Sichuan Science and Technology Program,No.2022ZYD0066(to YS)a grant from Chinese Academy of Medical Science,No.2019-12M-5-032(to YS)the Fundamental Research Funds for the Central Universities,No.ZYGX2021YGLH219(to KC)。
文摘Glaucoma is a leading cause of irreve rsible blindness wo rldwide,and previous studies have shown that,in addition to affecting the eyes,it also causes abnormalities in the brain.However,it is not yet clear how the primary visual cortex(V1)is altered in glaucoma.This study used DBA/2J mice as a model for spontaneous secondary glaucoma.The aim of the study was to compare the electrophysiological and histomorphological chara cteristics of neurons in the V1between 9-month-old DBA/2J mice and age-matched C57BL/6J mice.We conducted single-unit recordings in the V1 of light-anesthetized mice to measure the visually induced responses,including single-unit spiking and gamma band oscillations.The morphology of layerⅡ/Ⅲneurons was determined by neuronal nuclear antigen staining and Nissl staining of brain tissue sections.Eighty-seven neurons from eight DBA/2J mice and eighty-one neurons from eight C57BL/6J mice were examined.Compared with the C57BL/6J group,V1 neurons in the DBA/2J group exhibited weaker visual tuning and impaired spatial summation.Moreove r,fewer neuro ns were observed in the V1 of DBA/2J mice compared with C57BL/6J mice.These findings suggest that DBA/2J mice have fewer neurons in the VI compared with C57BL/6J mice,and that these neurons have impaired visual tuning.Our findings provide a better understanding of the pathological changes that occur in V1 neuron function and morphology in the DBA/2J mouse model.This study might offer some innovative perspectives regarding the treatment of glaucoma.
基金the National Natural Science Foundation of China(81973221)National Natural Science Foundation for Young Scientists of China(81603647)+2 种基金the Women and Children Health Talent Project of Jiangsu Province(FRC201785)the Chinese Clinical Medicine Innovation Center of Obstetrics,Gynecology,and Reproduction in Jiangsu Province(ZX202102)the Women and Children Health Science Foundation of Jiangsu Province(F202206).
文摘Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, traditional Chinese medicine has become a feasible alternative for certain diseases, including recurrent spontaneous abortion. However, because of the complex composition of the traditional Chinese medicine formula, its action mechanism remains unclear. Methods: We selected two isolated active ingredients (RAMP and baicalin) from the traditional Chinese medicine formula and used an abortion-prone CBA/J × DBA/2 model to simulate human RSA and compared the changes in fetal resorption rate, Treg cell percentage, and relevant cytokines before and after combination therapy. In addition, The mechanisms were preliminarily discussed using in vitro differentiation models. Results: In CBA/J × DBA/2 abortion-prone mice, the combination therapy resulted in a lower embryo resorption rate compared to that obtained with individual delivery of either RAMP or baicalin, thereby playing an embryo-protective role through the increase in Treg cells for the maintenance of maternal-fetal immune tolerance. In in vitro primary cell differentiation experiments, the concentration of Treg cells significantly increased from 11% to 17.9% after the combination therapy compared to that of the single administration group. Conclusion: the synergistic effects of RAMP and baicalin were responsible for Treg differentiation. The present study provides a solid basis for improving the applicability of traditional Chinese herbs in the treatment of recurrent spontaneous abortion.
文摘背景研究表明青光眼的眼压增高与转化生长因子-β（TGF-β）促进小梁网细胞外基质的堆积以及黏附分子CD44导致房水排出阻力增加有关。传统中药青光安颗粒剂是中医临床上常用的降跟压药物，其是否通过小梁网通路发挥作用尚不清楚。目的研究青光安颗粒剂对自发性青光眼模型DBA／2J小鼠房水动力学的影响及其作用机制。方法选取lO只眼压正常的3月龄雌性DBA／2J小鼠作为对照组，采用计算机随机数字分配法将20只9月龄自发性眼压升高的DBA／2J小鼠随机分为高眼压组和青光安组，青光安组小鼠采用2．5g／kg复方青光安颗粒剂灌胃，每天2次，连续15d，对照组和高眼压组小鼠以相同剂量生理盐水灌胃。采用经前房注入／抽吸系统进行眼压测量，分别以2．5、5．0μl／min的速率继续灌注，计算房水流畅系数（c）值和房水流出阻力（R）值。将3月龄DBA／2J小鼠60只按计算机随机数字分配法分为高剂量青光安组、中剂量青光安组、低剂量青光安组和空白对照组，每组各15只，分别用25．00、12．50和6．25g／kg青光安颗粒剂灌胃，空白对照组用等容量生理盐水灌胃，给药后7d麻醉条件下提取各组小鼠含青光安药物血清和空白对照动物血清，然后收集各组小鼠含小梁网巩膜组织进行培养，采用纤连蛋白（FN）、层黏连蛋白（LN）和神经元特异性烯醇化酶（NSE）免疫组织化学染色鉴定小梁网细胞。终质量浓度为0、5、10、20、50和100ng／ml的TGF-β处理小梁网细胞24h，采用MTT比色法检测各组细胞活性；用不同质量浓度含药血清培养经20ng／mlTGF-β处理的小梁细胞，分别于培养后24、48和72h采用ELISA法检测细胞上清液中TGF-β2受体质量浓度，采用Westernblot法检测各组小梁网细胞中CD44蛋白的相对表达量。结果对照组、高眼压组和青光安组小鼠在2．5td／min和5．0μl／min灌注速率下眼压明显不同，高眼压组小鼠眼压值明显高于青光安组和对照组，青光安组小鼠c值较高眼压组明显降低（1．08±O．36％2．35±1．34），R值明显增加（1．05±O．47 vs 0．64±0．55），差异均有统计学意义（均P〈O．01）。原代培养的细胞呈长梭形，FN、LN和NSE均呈阳性表达。5、10和20ng／mlTGF．p处理组细胞活力值明显低于0ng／mlTGF-β处理组细胞，差异均有统计学意义（均P〈0．05）。与空白对照组比较，TGF．母组小梁网细胞上清液中TGF-β2受体质量浓度和细胞中CD44蛋白相对表达量均明显升高，差异均有统计学意义（均P〈O．01）；细胞培养后24、48和72hTGF-β高剂量含药血清组细胞上清液中TGF-β2受体质量浓度和细胞中CD44相对表达量均明显低于TGF-β组和TGF-B＋低剂量含药血清组，差异均有统计学意义（均P〈0．01）。结论青光安颗粒剂通过影响房水动力学因素达到降低青光眼眼压的作用。青光安含药血清能明显降低体外培养的鼠小梁网细胞中TGF-β2受体和CD44的表达。
