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
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
目的:探究卵巢癌(ovarian cancer,OV)浸润性CD4^(+)调节性T细胞(regulatory T cell,Treg)的脂质摄取与积累情况,及其与CD4^(+)Treg中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)和细胞毒性T淋巴细胞相关蛋白-4(cytotoxi...目的:探究卵巢癌(ovarian cancer,OV)浸润性CD4^(+)调节性T细胞(regulatory T cell,Treg)的脂质摄取与积累情况,及其与CD4^(+)Treg中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)和细胞毒性T淋巴细胞相关蛋白-4(cytotoxic Tlymphocyte associated protein 4,CTLA-4)表达的相关性。方法:采用亲脂性荧光染料BODIPY^(TM)493/503和荧光脂肪酸探针BODIPY^(TM)500/510 C1 C12分别检测OV组织中的或与不同OV细胞系(ES-2、SKOV3、CAOV3)上清液共培养的人源CD4^(+)Treg的细胞内脂质含量和脂质摄取能力;使用脂肪酸氧化抑制剂(Etomoxir)、脂肪酸合成抑制剂(C75)和脂肪酸摄取抑制剂磺基-N-琥珀酰亚胺油酸酯(sulfo-N-succinimidyl oleate,SSO)干预脂质代谢;通过流式细胞术分析CD4^(+)Treg上免疫抑制分子PD-1和CTLA-4的表达。结果:OV组织CD4^(+)Treg相比传统CD4^(+)T细胞表现出更高的脂质含量和脂质摄取能力(P均<0.01)。在体外实验中,与基础培养基相比,OV细胞培养上清可显著提升CD4^(+)Treg的胞内脂质含量与脂质摄取能力(P均<0.05),其中CAOV3来源的上清作用最为显著。此外,CAOV3上清液还能提升CD4^(+)Treg中PD-1与CTLA-4的表达(P<0.05)。CD4^(+)Treg的脂质积累随CAOV3上清浓度增加呈剂量依赖性上升(P<0.05),且其对胞外荧光脂肪酸类似物的摄取能力具有浓度依赖性(P<0.05)。脂肪酸摄取抑制剂SSO可有效逆转CAOV3上清液诱导的CD4^(+)Treg脂质积累及PD-1、CTLA-4的高表达(P<0.05);而脂肪酸氧化抑制剂Etomoxir与合成抑制剂C75则无显著影响。结论:OV微环境通过促进CD4^(+)Treg的脂质摄取,提高细胞内脂质含量,促进其免疫抑制分子PD-1和CTLA-4表达。靶向脂肪酸摄取途径可能是逆转OV中Treg介导的免疫抑制的潜在策略。展开更多
旨在制备猫PD-L1抗原并筛选单抗,探讨其在犬、猫肿瘤检测和诊断中的应用,同时制备猫源化嵌合单链抗体,为猫肿瘤的治疗提供基础。本研究通过PCR扩增猫PD-L1基因,构建原核表达重组质粒PET-MPD-L1,通过在宿主BL21(DE3)中表达制备PD-L1抗原...旨在制备猫PD-L1抗原并筛选单抗,探讨其在犬、猫肿瘤检测和诊断中的应用,同时制备猫源化嵌合单链抗体,为猫肿瘤的治疗提供基础。本研究通过PCR扩增猫PD-L1基因,构建原核表达重组质粒PET-MPD-L1,通过在宿主BL21(DE3)中表达制备PD-L1抗原,免疫小鼠后细胞融合,ELISA方法进行杂交瘤细胞筛选,3次亚克隆后建立MPD-L1杂交瘤细胞系。测定杂交瘤细胞上清、腹水效价、抗体亚类、Western blot和免疫组织化学(IHC)特性。扩增轻、重链可变区和猫IgG1 Fc基因,降落PCR将重链可变区(Variable region of the Heavy chain,VH)和轻链可变区(Variable region of the Light chain,VL)拼接成scFv,进一步拼接出scFv-fFc,构建pFast-MPD-L1-scFv和pFast-MPD-L1-scFv-fFc重组质粒,在昆虫杆状病毒表达系统中表达。结果表明,猫PD-L1在上清中大量表达且纯度较好,杂交瘤细胞2A5A2F4D7分泌的抗体属于IgG 2a,轻链为κ亚型,细胞上清、腹水抗体效价分别为1∶3200和1∶102400,Western blot结果表明,该单抗能与表达的猫PD-L1反应,IHC结果显示,该单抗可以识别犬猫肿瘤组织细胞表达的PD-L1,可以反映PD-L1在肿瘤组织中的表达情况。VH和VL与IMGT数据库中相应基因序列相符,具有4个框架区和3个高变区,猫源化嵌合单链抗体在昆虫杆状病毒表达系统中成功表达。综上,本研究制备的猫PD-L1单克隆抗体在犬猫肿瘤检测和预后中具有一定的价值,猫源化单链抗体的成功制备为其在犬猫肿瘤病治疗中的应用打下了基础。展开更多
Examining carbonate dissolution kinetics at mineral-water interface is crucial to understand numerous environmental and geochemical processes,including global carbon cycling,CO_(2)sequestration in deep geological rese...Examining carbonate dissolution kinetics at mineral-water interface is crucial to understand numerous environmental and geochemical processes,including global carbon cycling,CO_(2)sequestration in deep geological reservoirs,and trace elements release in terrestrial and aquatic environments.Here we explored the effect of circumneutral to alkaline pH solutions(pH 6-11)on dissolution kinetics of pure dolomite and Ca and Mg release stoichiometry in flow-through reactor experiments at 25±1℃.Results revealed that the dolomite dissolution rates obtained from effluent Ca and Mg concentrations(R_(Ca)and R_(Mg)in mol/cm^(2)/s)were dependent on input solution pH and HCO_(3)^(-)log activity.The pH dependence of dissolution rates showed two distinct trends,i.e.,at circumneutral pH ranging between 6 and 8,the dissolution rate decreased with increasing pH,with minimum rate at pH 8.While in the highly alkaline pH range(pH 9-11),the dolomite dissolution rate increased with an increasing pH.Irrespective of the input pH,the dolomite dissolution rates indicated a reverse relationship with HCO_(3)^(-)log activity,with the lowest dissolution rate(R Ca=3.80×10^(-12)mol/cm^(2)/s)at pH 8 where HCO_(3)^(-)log activity attained the highest value(-3.957).The lower R Ca and R Mg obtained at pH 8 compared to all the other pH could possibly be attributed to an inhibition caused by high HCO_(3)^(-)log activity in solution at this pH.Dolomite dissolution rates were non-stoichiometric at all the experimental pH values,showing higher preferential Ca over Mg release(R_(Ca)>R_(Mg))whereas an opposite trend was observed at pH 8,with R_(Ca)<R_(Mg)at the steady state.Saturation index values calculated using geochemical speciation modelling were positive for Mg-bearing minerals(brucite,dolomite,artinite)at alkaline pH of 10-11,indicating favourable conditions for their precipitation under studied conditions.This study provides insights on the significance of log ion activities of HCO_(3)^(-)and Me-OH^(+)under varying pH for elucidating the dissolution mechanism of dolomite in circumneutral to alkaline aqueous environments.展开更多
Nanofiltration(NF) technology,with its capacity for nanoscale filtration and controllable selectivity,holds significant promise in diverse applications.However,the current upper bound of permeance and selectivity of N...Nanofiltration(NF) technology,with its capacity for nanoscale filtration and controllable selectivity,holds significant promise in diverse applications.However,the current upper bound of permeance and selectivity of NF membranes is intrinsically constrained by the morphology and structure of the polyamide(PA) selective layer.This issue arises because NF membranes typically exhibit relatively smooth nodular structures,which theoretically impede efficient water transport.In this study,we enhanced the formation of nanobubbles by synergistically regulating with surfactant and low temperatures,resulting in the fabrication of PA NF membranes with a crumpled morphology.We observed that lower temperatures promote enhanced gas solubility in the aqueous phase,facilitating increased nanobubble formation through the foaming effect of surfactant sodium dodecylbenzene sulfonate(SDBS).Consequently,this resulted in the creation of PA NF membranes with more crumpled structures and superior performance,with pure water permeance reaching 36.25 ± 0.42 L m^(-2)h^(-1)bar^(-1),representing an improvement of 14.47 L m^(-2)h^(-1)bar^(-1)compared to the control group.Additionally,it maintains a high Na_(2)SO_(4) rejection rate of97.00 % ± 0.58 %.The PA NF membranes produced by eliminating nanobubbles and free interfaces exhibited a smooth structure,whereas introducing nanobubbles(through Na HCO_(3) addition,N_(2) pressurization,and ultrasonication) resulted in the formation of crumpled membranes.This emphasized that the large amount of nanobubbles generated by SDBS and low temperature in the interfacial process played a critical role in shaping crumpled PA NF membranes and enhancing membrane performance.This approach has the potential to provide valuable insights into customizing the structural design of TFC PA NF membranes,contributing to further advancements in this field.展开更多
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。
文摘目的:探究卵巢癌(ovarian cancer,OV)浸润性CD4^(+)调节性T细胞(regulatory T cell,Treg)的脂质摄取与积累情况,及其与CD4^(+)Treg中程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)和细胞毒性T淋巴细胞相关蛋白-4(cytotoxic Tlymphocyte associated protein 4,CTLA-4)表达的相关性。方法:采用亲脂性荧光染料BODIPY^(TM)493/503和荧光脂肪酸探针BODIPY^(TM)500/510 C1 C12分别检测OV组织中的或与不同OV细胞系(ES-2、SKOV3、CAOV3)上清液共培养的人源CD4^(+)Treg的细胞内脂质含量和脂质摄取能力;使用脂肪酸氧化抑制剂(Etomoxir)、脂肪酸合成抑制剂(C75)和脂肪酸摄取抑制剂磺基-N-琥珀酰亚胺油酸酯(sulfo-N-succinimidyl oleate,SSO)干预脂质代谢;通过流式细胞术分析CD4^(+)Treg上免疫抑制分子PD-1和CTLA-4的表达。结果:OV组织CD4^(+)Treg相比传统CD4^(+)T细胞表现出更高的脂质含量和脂质摄取能力(P均<0.01)。在体外实验中,与基础培养基相比,OV细胞培养上清可显著提升CD4^(+)Treg的胞内脂质含量与脂质摄取能力(P均<0.05),其中CAOV3来源的上清作用最为显著。此外,CAOV3上清液还能提升CD4^(+)Treg中PD-1与CTLA-4的表达(P<0.05)。CD4^(+)Treg的脂质积累随CAOV3上清浓度增加呈剂量依赖性上升(P<0.05),且其对胞外荧光脂肪酸类似物的摄取能力具有浓度依赖性(P<0.05)。脂肪酸摄取抑制剂SSO可有效逆转CAOV3上清液诱导的CD4^(+)Treg脂质积累及PD-1、CTLA-4的高表达(P<0.05);而脂肪酸氧化抑制剂Etomoxir与合成抑制剂C75则无显著影响。结论:OV微环境通过促进CD4^(+)Treg的脂质摄取,提高细胞内脂质含量,促进其免疫抑制分子PD-1和CTLA-4表达。靶向脂肪酸摄取途径可能是逆转OV中Treg介导的免疫抑制的潜在策略。
文摘旨在制备猫PD-L1抗原并筛选单抗,探讨其在犬、猫肿瘤检测和诊断中的应用,同时制备猫源化嵌合单链抗体,为猫肿瘤的治疗提供基础。本研究通过PCR扩增猫PD-L1基因,构建原核表达重组质粒PET-MPD-L1,通过在宿主BL21(DE3)中表达制备PD-L1抗原,免疫小鼠后细胞融合,ELISA方法进行杂交瘤细胞筛选,3次亚克隆后建立MPD-L1杂交瘤细胞系。测定杂交瘤细胞上清、腹水效价、抗体亚类、Western blot和免疫组织化学(IHC)特性。扩增轻、重链可变区和猫IgG1 Fc基因,降落PCR将重链可变区(Variable region of the Heavy chain,VH)和轻链可变区(Variable region of the Light chain,VL)拼接成scFv,进一步拼接出scFv-fFc,构建pFast-MPD-L1-scFv和pFast-MPD-L1-scFv-fFc重组质粒,在昆虫杆状病毒表达系统中表达。结果表明,猫PD-L1在上清中大量表达且纯度较好,杂交瘤细胞2A5A2F4D7分泌的抗体属于IgG 2a,轻链为κ亚型,细胞上清、腹水抗体效价分别为1∶3200和1∶102400,Western blot结果表明,该单抗能与表达的猫PD-L1反应,IHC结果显示,该单抗可以识别犬猫肿瘤组织细胞表达的PD-L1,可以反映PD-L1在肿瘤组织中的表达情况。VH和VL与IMGT数据库中相应基因序列相符,具有4个框架区和3个高变区,猫源化嵌合单链抗体在昆虫杆状病毒表达系统中成功表达。综上,本研究制备的猫PD-L1单克隆抗体在犬猫肿瘤检测和预后中具有一定的价值,猫源化单链抗体的成功制备为其在犬猫肿瘤病治疗中的应用打下了基础。
基金funding enabled and organized by CAUL and its Member Institutionsby COMSTEQ-TWAS research grant 2018(18-268 RG/EAS/AS_C)。
文摘Examining carbonate dissolution kinetics at mineral-water interface is crucial to understand numerous environmental and geochemical processes,including global carbon cycling,CO_(2)sequestration in deep geological reservoirs,and trace elements release in terrestrial and aquatic environments.Here we explored the effect of circumneutral to alkaline pH solutions(pH 6-11)on dissolution kinetics of pure dolomite and Ca and Mg release stoichiometry in flow-through reactor experiments at 25±1℃.Results revealed that the dolomite dissolution rates obtained from effluent Ca and Mg concentrations(R_(Ca)and R_(Mg)in mol/cm^(2)/s)were dependent on input solution pH and HCO_(3)^(-)log activity.The pH dependence of dissolution rates showed two distinct trends,i.e.,at circumneutral pH ranging between 6 and 8,the dissolution rate decreased with increasing pH,with minimum rate at pH 8.While in the highly alkaline pH range(pH 9-11),the dolomite dissolution rate increased with an increasing pH.Irrespective of the input pH,the dolomite dissolution rates indicated a reverse relationship with HCO_(3)^(-)log activity,with the lowest dissolution rate(R Ca=3.80×10^(-12)mol/cm^(2)/s)at pH 8 where HCO_(3)^(-)log activity attained the highest value(-3.957).The lower R Ca and R Mg obtained at pH 8 compared to all the other pH could possibly be attributed to an inhibition caused by high HCO_(3)^(-)log activity in solution at this pH.Dolomite dissolution rates were non-stoichiometric at all the experimental pH values,showing higher preferential Ca over Mg release(R_(Ca)>R_(Mg))whereas an opposite trend was observed at pH 8,with R_(Ca)<R_(Mg)at the steady state.Saturation index values calculated using geochemical speciation modelling were positive for Mg-bearing minerals(brucite,dolomite,artinite)at alkaline pH of 10-11,indicating favourable conditions for their precipitation under studied conditions.This study provides insights on the significance of log ion activities of HCO_(3)^(-)and Me-OH^(+)under varying pH for elucidating the dissolution mechanism of dolomite in circumneutral to alkaline aqueous environments.
基金the National Natural Science Foundation of China (Nos.52430001,52470091,52200108) for the financial support。
文摘Nanofiltration(NF) technology,with its capacity for nanoscale filtration and controllable selectivity,holds significant promise in diverse applications.However,the current upper bound of permeance and selectivity of NF membranes is intrinsically constrained by the morphology and structure of the polyamide(PA) selective layer.This issue arises because NF membranes typically exhibit relatively smooth nodular structures,which theoretically impede efficient water transport.In this study,we enhanced the formation of nanobubbles by synergistically regulating with surfactant and low temperatures,resulting in the fabrication of PA NF membranes with a crumpled morphology.We observed that lower temperatures promote enhanced gas solubility in the aqueous phase,facilitating increased nanobubble formation through the foaming effect of surfactant sodium dodecylbenzene sulfonate(SDBS).Consequently,this resulted in the creation of PA NF membranes with more crumpled structures and superior performance,with pure water permeance reaching 36.25 ± 0.42 L m^(-2)h^(-1)bar^(-1),representing an improvement of 14.47 L m^(-2)h^(-1)bar^(-1)compared to the control group.Additionally,it maintains a high Na_(2)SO_(4) rejection rate of97.00 % ± 0.58 %.The PA NF membranes produced by eliminating nanobubbles and free interfaces exhibited a smooth structure,whereas introducing nanobubbles(through Na HCO_(3) addition,N_(2) pressurization,and ultrasonication) resulted in the formation of crumpled membranes.This emphasized that the large amount of nanobubbles generated by SDBS and low temperature in the interfacial process played a critical role in shaping crumpled PA NF membranes and enhancing membrane performance.This approach has the potential to provide valuable insights into customizing the structural design of TFC PA NF membranes,contributing to further advancements in this field.
