[Objective]To systematically isolate and purify the polysaccharide from the mycelium of Streptomyces rochei D74(SRP),elucidate its fine structure,and evaluate the effect of the purified polysaccharide fraction on the ...[Objective]To systematically isolate and purify the polysaccharide from the mycelium of Streptomyces rochei D74(SRP),elucidate its fine structure,and evaluate the effect of the purified polysaccharide fraction on the growth of Salvia miltiorrhiza hairy roots and the biosynthesis of tanshinones,along with the underlying mechanism.[Methods]The crude polysaccharide was extracted using hot water,which was followed by ethanol precipitation and deproteinization via the Sevag method.Further purification was performed using DEAE-52 anionexchange chromatography and Sephadex G-100 gel filtration chromatography.The physicochemical properties and structural features of the main active fraction,SRP-W-2,were systematically characterized by Fourier transform infrared spectroscopy(FTIR),high performance liquid chromatography-mass spectrometry(HPLC-MS),and nuclear magnetic resonance(NMR).The effects of SRP-W-2 on hairy root growth and the biosynthesis of tanshinones were assessed by measuring biomass,tanshinone content,and the expression levels of key biosynthetic genes.[Results]SRP-W-2 was obtained with a yield of 2.41%.It was primarily composed of glucose and galactose at a molar ratio of 12.53:1.Structural analysis revealed that the backbone of SRP-W-2 consisted of→4)-α-D-Glcp-(1→and→4)-α-D-Galp-(1→residues,with branching points at→4,6)-α-D-Glcp-(1→and→4,6)-α-D-Galp-(1→.The side chain was identified asα-D-Glcp-(1→4)-α-DGlcp-(1→.Bioactivity assays demonstrated that SRP-W-2 significantly enhanced both the biomass of S.miltiorrhiza hairy roots and the accumulation of tanshinones.After 15 d of treatment with 50 mg/L SRP-W-2,the dry weight of the hairy roots increased by 37.52%.Meanwhile,the content of cryptotanshinone(CT),dihydrotanshinone I(DT-I),tanshinone I(T-I),and tanshinone IIA(TIIA)was increased by 19.0-fold,6.4-fold,2.8-fold,and 4.8-fold,respectively.Gene expression analysis further indicated that SRP-W-2 up-regulated key genes involved in the tanshinone biosynthetic pathway,including HMGR,DXS,DXR,and GGPPS.[Conclusion]The polysaccharide fraction SRP-W-2 from S.rochei D74 simultaneously promoted the growth of S.miltiorrhiza hairy roots and the biosynthesis of tanshinones,demonstrating its potential as an effective elicitor.This study provided a new strategy for the utilization and development of S.miltiorrhiza resources.展开更多
目的探讨CMTM6、PD-L1在子宫内膜异位症(endometriosis,EM)中的表达及意义。方法收集50例EM患者腹腔镜下手术切除的卵巢异位子宫内膜组织、刮除的在位子宫内膜组织(实验组)以及30例非EM患者刮除的在位子宫内膜组织(对照组),采用免疫组...目的探讨CMTM6、PD-L1在子宫内膜异位症(endometriosis,EM)中的表达及意义。方法收集50例EM患者腹腔镜下手术切除的卵巢异位子宫内膜组织、刮除的在位子宫内膜组织(实验组)以及30例非EM患者刮除的在位子宫内膜组织(对照组),采用免疫组化法检测CMTM6、PD-L1的表达水平。收集实验组相关临床病理资料,包括患者年龄、体重指数(body mass index,BMI)、病灶大小、血清CA-125浓度及rAFS评分等,采用Spearman系数法分析CMTM6、PD-L1的表达与上述病理参数的相关性。培养人EM上皮细胞系12Z,采用Western blot及免疫蛋白共沉淀法检测CMTM6、PD-L1的表达水平及相互作用。结果CMTM6、PD-L1在EM患者中高表达,EM患者异位、在位子宫内膜中的表达水平高于非EM患者在位子宫内膜组织(P<0.05),CMTM6、PD-L1表达共同定位于异位、在位子宫内膜组织上皮细胞质、细胞膜中,且两种蛋白表达呈正相关(P<0.05)。患者年龄、BMI、病灶大小、血清CA-125浓度及rAFS评分与CMTM6、PD-L1的表达呈正相关(P<0.01)。CMTM6、PD-L1蛋白在12Z中表达,且CMTM6和PD-L能够相互结合。结论CMTM6、PD-L1在EM患者中高表达,CMTM6、PDL1的表达水平与EM病变严重程度相关,CMTM6、PD-L1在EM中能相互结合共同参与EM的发生、发展,CMTM6、PD-L1或可成为EM的潜在生物学标志物,为EM的临床诊治提供新靶点。展开更多
目的探索酮还原酶家族1成员C3(aldo-keto reductase family 1 member C3,AKR1C3)对乳腺癌恶性细胞生物学行为的干预作用及对程序性细胞死亡蛋白/程序性死亡-配体1(programmed cell death protein1/programmed death-ligand1,PD-1/PD-L)...目的探索酮还原酶家族1成员C3(aldo-keto reductase family 1 member C3,AKR1C3)对乳腺癌恶性细胞生物学行为的干预作用及对程序性细胞死亡蛋白/程序性死亡-配体1(programmed cell death protein1/programmed death-ligand1,PD-1/PD-L)通路的影响。方法把MCF-7人乳腺癌细胞中NC组和AKR1C3组分别转染空质粒和AKR1C3质粒,采用MTT法检测转染后24 h、48 h、72 h细胞活力;采用流式细胞技术测定各组细胞的存活率以及早期、晚期凋亡比例;通过Transwell实验对各组细胞的迁移和侵袭能力进行检测;通过Western blot检测各组细胞PD-1、PD-L1、蛋白激酶B(protein kinase b,AKT)蛋白表达水平。使用C57BL/6小鼠构建荷瘤模型,将采用人乳腺癌MCF-7细胞转染NC质粒和AKR1C3质粒进行细胞荷瘤,每3 d测量瘤体积,持续21 d,绘制两组小鼠肿瘤生长曲线,并于实验终点测量肿瘤质量。结果相较于NC组,AKR1C3组细胞活力降低(P<0.05),并且具有时间依赖效应(P<0.05),迁移和侵袭能力降低(P<0.05),早期凋亡和晚期凋亡比例升高(P<0.05),PD-1、PD-L1、AKT蛋白表达水平降低(P<0.05)。动物实验表明,AKR1C3组小鼠肿瘤体积降低,肿瘤质量下降(P<0.05)。结论AKR1C3可以抑制人乳腺癌细胞恶性生物学行为,抑制PD-1/PDL1信号通路蛋白表达。展开更多
目的约70%接受程序性细胞死亡蛋白1(Programmed Cell Death Protein-1,PD-1)及其配体(Programmed Cell Death Ligand 1,PD-L1)抑制剂治疗的患者会出现免疫相关不良事件(Immune-Related Adverse Events,irAEs)。然而,PD-1/PD-L1抑制剂导...目的约70%接受程序性细胞死亡蛋白1(Programmed Cell Death Protein-1,PD-1)及其配体(Programmed Cell Death Ligand 1,PD-L1)抑制剂治疗的患者会出现免疫相关不良事件(Immune-Related Adverse Events,irAEs)。然而,PD-1/PD-L1抑制剂导致irAEs的机制仍不清楚。方法本研究中,我们从公开的全基因组关联研究(Genome-Wide Association Studies,GWAS)中收集了涉及欧洲血统个体的数据。采用双样本孟德尔随机化(Two-Sample Mendelian Randomization,TSMR)方法探讨了PD-1/PD-L1与免疫相关不良事件之间的因果关系,以及731种免疫细胞、1400种代谢物和41种炎症细胞因子的介导效应。此外,我们还通过敏感性分析来验证结果的异质性和多效性,以加强结果的稳健性。结果我们确定了PD-1/PD-L1与免疫相关不良事件之间的九种因果关系,其中我们发现在常见的irAEs中,PD-1/PD-L1与胃肠道疾病和内分泌腺疾病的水平增加相关,而在其他irAEs中,呼吸系统疾病、精神疾病和免疫系统疾病与PD-1/PD-L1相关。在反向分析中,我们发现抑郁症与PD-L1之间存在显著的因果关系。此外,我们还发现PD-1可通过N-甲基羟脯氨酸水平影响过敏性鼻炎,而PD-L1可通过3-甲基己二酸水平和羟基棕榈酰鞘磷脂(d18:1/16:0(OH))水平影响哮喘。结论MR分析揭示了PD-1/PD-L1与免疫相关不良事件之间的双向因果关系,而这种关系可能是由代谢物介导的。展开更多
构建使用了PD-1抑制剂的肿瘤患者出现甲状腺功能障碍的风险预测模型,分析使用PD-1肿瘤抑制剂导致的甲状腺功能障碍的相关风险因素,设计监测预警系统。选取2020年—2023年广西医科大学附属肿瘤医院1225例使用PD-1抑制剂肿瘤患者的临床资...构建使用了PD-1抑制剂的肿瘤患者出现甲状腺功能障碍的风险预测模型,分析使用PD-1肿瘤抑制剂导致的甲状腺功能障碍的相关风险因素,设计监测预警系统。选取2020年—2023年广西医科大学附属肿瘤医院1225例使用PD-1抑制剂肿瘤患者的临床资料,包括人口学特征、既往史、实验室检测等63个变量。本文选取相关性前10/20/30/40/50/60个变量的4种传统机器学习模型进行性能比较。通过F1分数、灵敏度、准确率、精确率、特异性曲线下面积(Area Under the Curve,AUC)评估以上预测模型的性能,并利用Shapley加性解释(Shapley Additive Explanation,SHAP)可视化解释本文的机器学习模型。与促甲状腺激素相关性排名前10的变量依次为:羟丁酸脱氢酶、乳酸脱氢酶、淋巴细胞绝对值、天门冬氨酸转移酶、钙离子、碱性磷酸酶、谷氨酰转肽酶、单核细胞绝对值、红细胞分布宽度SD、胆碱酯酶。建立了使用PD-1抑制剂的肿瘤患者出现甲状腺功能障碍的风险预测模型,并在全局解释和局部解释的层面上分别作出模型预测结果影响的解释。展开更多
文摘[Objective]To systematically isolate and purify the polysaccharide from the mycelium of Streptomyces rochei D74(SRP),elucidate its fine structure,and evaluate the effect of the purified polysaccharide fraction on the growth of Salvia miltiorrhiza hairy roots and the biosynthesis of tanshinones,along with the underlying mechanism.[Methods]The crude polysaccharide was extracted using hot water,which was followed by ethanol precipitation and deproteinization via the Sevag method.Further purification was performed using DEAE-52 anionexchange chromatography and Sephadex G-100 gel filtration chromatography.The physicochemical properties and structural features of the main active fraction,SRP-W-2,were systematically characterized by Fourier transform infrared spectroscopy(FTIR),high performance liquid chromatography-mass spectrometry(HPLC-MS),and nuclear magnetic resonance(NMR).The effects of SRP-W-2 on hairy root growth and the biosynthesis of tanshinones were assessed by measuring biomass,tanshinone content,and the expression levels of key biosynthetic genes.[Results]SRP-W-2 was obtained with a yield of 2.41%.It was primarily composed of glucose and galactose at a molar ratio of 12.53:1.Structural analysis revealed that the backbone of SRP-W-2 consisted of→4)-α-D-Glcp-(1→and→4)-α-D-Galp-(1→residues,with branching points at→4,6)-α-D-Glcp-(1→and→4,6)-α-D-Galp-(1→.The side chain was identified asα-D-Glcp-(1→4)-α-DGlcp-(1→.Bioactivity assays demonstrated that SRP-W-2 significantly enhanced both the biomass of S.miltiorrhiza hairy roots and the accumulation of tanshinones.After 15 d of treatment with 50 mg/L SRP-W-2,the dry weight of the hairy roots increased by 37.52%.Meanwhile,the content of cryptotanshinone(CT),dihydrotanshinone I(DT-I),tanshinone I(T-I),and tanshinone IIA(TIIA)was increased by 19.0-fold,6.4-fold,2.8-fold,and 4.8-fold,respectively.Gene expression analysis further indicated that SRP-W-2 up-regulated key genes involved in the tanshinone biosynthetic pathway,including HMGR,DXS,DXR,and GGPPS.[Conclusion]The polysaccharide fraction SRP-W-2 from S.rochei D74 simultaneously promoted the growth of S.miltiorrhiza hairy roots and the biosynthesis of tanshinones,demonstrating its potential as an effective elicitor.This study provided a new strategy for the utilization and development of S.miltiorrhiza resources.
文摘目的探讨CMTM6、PD-L1在子宫内膜异位症(endometriosis,EM)中的表达及意义。方法收集50例EM患者腹腔镜下手术切除的卵巢异位子宫内膜组织、刮除的在位子宫内膜组织(实验组)以及30例非EM患者刮除的在位子宫内膜组织(对照组),采用免疫组化法检测CMTM6、PD-L1的表达水平。收集实验组相关临床病理资料,包括患者年龄、体重指数(body mass index,BMI)、病灶大小、血清CA-125浓度及rAFS评分等,采用Spearman系数法分析CMTM6、PD-L1的表达与上述病理参数的相关性。培养人EM上皮细胞系12Z,采用Western blot及免疫蛋白共沉淀法检测CMTM6、PD-L1的表达水平及相互作用。结果CMTM6、PD-L1在EM患者中高表达,EM患者异位、在位子宫内膜中的表达水平高于非EM患者在位子宫内膜组织(P<0.05),CMTM6、PD-L1表达共同定位于异位、在位子宫内膜组织上皮细胞质、细胞膜中,且两种蛋白表达呈正相关(P<0.05)。患者年龄、BMI、病灶大小、血清CA-125浓度及rAFS评分与CMTM6、PD-L1的表达呈正相关(P<0.01)。CMTM6、PD-L1蛋白在12Z中表达,且CMTM6和PD-L能够相互结合。结论CMTM6、PD-L1在EM患者中高表达,CMTM6、PDL1的表达水平与EM病变严重程度相关,CMTM6、PD-L1在EM中能相互结合共同参与EM的发生、发展,CMTM6、PD-L1或可成为EM的潜在生物学标志物,为EM的临床诊治提供新靶点。
文摘目的约70%接受程序性细胞死亡蛋白1(Programmed Cell Death Protein-1,PD-1)及其配体(Programmed Cell Death Ligand 1,PD-L1)抑制剂治疗的患者会出现免疫相关不良事件(Immune-Related Adverse Events,irAEs)。然而,PD-1/PD-L1抑制剂导致irAEs的机制仍不清楚。方法本研究中,我们从公开的全基因组关联研究(Genome-Wide Association Studies,GWAS)中收集了涉及欧洲血统个体的数据。采用双样本孟德尔随机化(Two-Sample Mendelian Randomization,TSMR)方法探讨了PD-1/PD-L1与免疫相关不良事件之间的因果关系,以及731种免疫细胞、1400种代谢物和41种炎症细胞因子的介导效应。此外,我们还通过敏感性分析来验证结果的异质性和多效性,以加强结果的稳健性。结果我们确定了PD-1/PD-L1与免疫相关不良事件之间的九种因果关系,其中我们发现在常见的irAEs中,PD-1/PD-L1与胃肠道疾病和内分泌腺疾病的水平增加相关,而在其他irAEs中,呼吸系统疾病、精神疾病和免疫系统疾病与PD-1/PD-L1相关。在反向分析中,我们发现抑郁症与PD-L1之间存在显著的因果关系。此外,我们还发现PD-1可通过N-甲基羟脯氨酸水平影响过敏性鼻炎,而PD-L1可通过3-甲基己二酸水平和羟基棕榈酰鞘磷脂(d18:1/16:0(OH))水平影响哮喘。结论MR分析揭示了PD-1/PD-L1与免疫相关不良事件之间的双向因果关系,而这种关系可能是由代谢物介导的。
文摘构建使用了PD-1抑制剂的肿瘤患者出现甲状腺功能障碍的风险预测模型,分析使用PD-1肿瘤抑制剂导致的甲状腺功能障碍的相关风险因素,设计监测预警系统。选取2020年—2023年广西医科大学附属肿瘤医院1225例使用PD-1抑制剂肿瘤患者的临床资料,包括人口学特征、既往史、实验室检测等63个变量。本文选取相关性前10/20/30/40/50/60个变量的4种传统机器学习模型进行性能比较。通过F1分数、灵敏度、准确率、精确率、特异性曲线下面积(Area Under the Curve,AUC)评估以上预测模型的性能,并利用Shapley加性解释(Shapley Additive Explanation,SHAP)可视化解释本文的机器学习模型。与促甲状腺激素相关性排名前10的变量依次为:羟丁酸脱氢酶、乳酸脱氢酶、淋巴细胞绝对值、天门冬氨酸转移酶、钙离子、碱性磷酸酶、谷氨酰转肽酶、单核细胞绝对值、红细胞分布宽度SD、胆碱酯酶。建立了使用PD-1抑制剂的肿瘤患者出现甲状腺功能障碍的风险预测模型,并在全局解释和局部解释的层面上分别作出模型预测结果影响的解释。
