目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunogl...目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunoglobulin new antigen receptor,VNAR)基因,将VNAR构建至噬菌体展示载体中,完成鲨鱼纳米抗体天然文库的构建。扩增天然噬菌体展示文库,采用抗原固相淘选策略,通过三轮“结合-洗脱-扩增”的淘选流程完成噬菌体展示文库的富集,通过噬菌体ELISA鉴定阳性克隆,并进行抗体序列比对,以确定靶向TfR1的鲨鱼纳米抗体序列。构建鲨鱼纳米抗体重组表达载体,利用大肠杆菌原核表达系统进行鲨鱼纳米抗体的诱导表达,ELISA检测鲨鱼纳米抗体与TfR1重组蛋白的结合能力,竞争性ELISA检测鲨鱼纳米抗体与转铁蛋白(transferrin,Tf)竞争受体结合表位的情况,免疫荧光技术检测鲨鱼纳米抗体与脑微血管内皮细胞bEnd.3的结合能力,血脑屏障体外实验模型检测鲨鱼纳米抗体的跨血脑屏障转运能力。结果成功构建了库容量为2.35×10^(9) CFU的鲨鱼纳米抗体天然文库,该文库具备良好的多样性。经淘选获得2株靶向TfR1的鲨鱼纳米抗体,均具有亲和活性,其中D1具有跨血脑屏障转运功能。结论本研究成功筛选出靶向TfR1且具有跨血脑屏障转运功能的鲨鱼纳米抗体,为中枢神经系统疾病治疗提供了新的跨血脑屏障药物递送载体。展开更多
Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District Peop...Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District People's Hospital,the First Affiliated Hospital of Chengdu Medical College and Mianyang Central Hospital between August 2015 and April 2018,115 patients with ovarian chocolate cyst underwent surgery during the same period,and intraoperative lesion tissue samples of the two groups were collected and enrolled in ovarian cancer group and ovarian chocolate cyst group respectively.The expressions levels of Numb,TFR1 gene as well as ovarian cancer-related proliferation,invasion and autophagy genes in ovarian lesions were detected.Pearson test was used to evaluate the correlation of Numb and TFR1 gene expression in ovarian cancer tissues with proliferation,invasion and autophagy activity of cancer cells.Results:Numb mRNA expression in lesions of ovarian cancer group was lower than that of ovarian chocolate cyst group whereas TFR1 mRNA expression was higher than that of ovarian chocolate cyst group;proliferation genes C3G,Clusterin,FUNDC1,PIWIL4 and TNFAIP8 mRNA expression were higher than those of ovarian chocolate cyst group whereas Let-7i mRNA expression was lower than that of ovarian chocolate cyst group;invasion genes BRMS1,XAV939 and OVCA1 mRMA expression were lower than those of ovarian chocolate cyst group whereas CUL4A and MTA1 mRNA expression were higher than those of ovarian chocolate cyst group;autophagy genes FTY720 and Beclin1 mRNA expression were lower than those of ovarian chocolate cyst group whereas NAC-1 mRNA expression was higher than that of ovarian chocolate cyst group.Correlation analysis showed that the Numb and TFR1 gene expression in ovarian cancer tissues were directly correlated with the proliferation,invasion and autophagy activity of cancer cells.Conclusion:The abnormal decrease of Numb gene expression and the abnormal increase of TFR1 gene expression in ovarian cancer tissues are directly correlated with the malignant biological behaviors of cancer cells.展开更多
Heat stroke(HS)is a severe medical emergency characterized by coagulation and high mortality due to organ injury.This study identifies a novel mechanism in which platelet ferroptosis,driven by transferrin receptor 1(T...Heat stroke(HS)is a severe medical emergency characterized by coagulation and high mortality due to organ injury.This study identifies a novel mechanism in which platelet ferroptosis,driven by transferrin receptor 1(Tfr1)palmitoylation,significantly contributes to liver injury in HS.Our findings reveal a strong inverse correlation between platelet count and organ damage,especially liver injury,as well as mortality rates.Using murine models,we demonstrate that inhibiting Tfr1-mediated ferroptosis in platelets mitigates thrombocytopenia and decreases Interleukin-1β(IL-1β)secretion,thereby improving liver function and survival outcomes.This research highlights Tfr1 palmitoylation as a critical factor in iron transport within platelets,with the palmitoylation inhibitor 2-bromopalmitate(2BP)effectively reducing total iron,Fe2+,lipid ROS,4-hydroxynonenal(4-HNE),and cell cytotoxicity under heat stress.These results suggest that targeting Tfr1 palmitoylation-dependent ferroptosis in platelets offers a novel therapeutic strategy for treating HS-induced thrombocytopenia and liver injury.展开更多
Mink enteritis virus(MEV),a member of the Protoparvovirus genus within the Parvovirinae subfamily,causes severe hemorrhagic enteritis and leukopenia in minks,thus representing a serious economic threat to mink farming...Mink enteritis virus(MEV),a member of the Protoparvovirus genus within the Parvovirinae subfamily,causes severe hemorrhagic enteritis and leukopenia in minks,thus representing a serious economic threat to mink farming.MEV,feline panleukopenia virus(FPV),and canine parvovirus(CPV)are closely related Protoparvovirus members;CPV and FPV infect hosts via binding to transferrin receptor 1(TfR1).However,the structural mechanism of MEV entry remains poorly understood.This study used cryo-electron microscopy(cryo-EM)and structural bioinformatics to determine the structure of MEV virus-like particles at 2.5 A resolution.The analysis revealed that MEV shares a highly conserved capsid architecture with CPV and FPV,particularly in theβ-barrel core and three-fold protrusions.We predict that MEV utilizes mink TfR1 for host entry,similar to CPV and FPV.Structural superposition and sequence alignment identified key residues in the MEV VP2 protein that are critical for mink TfR1 interaction.These findings provide structural and evolutionary insights into MEV host tropism,offering valuable information for efforts to mitigate the zoonotic risks of mammalian parvoviruses.展开更多
文摘目的开发靶向转铁蛋白受体1(transferrin receptor 1,TfR1)且具有跨血脑屏障转运功能的鲨鱼纳米抗体。方法提取未经靶抗原免疫的条纹斑竹鲨外周血单个核细胞总RNA,通过PCR技术扩增获得鲨鱼抗体的重链可变区(variable domain of immunoglobulin new antigen receptor,VNAR)基因,将VNAR构建至噬菌体展示载体中,完成鲨鱼纳米抗体天然文库的构建。扩增天然噬菌体展示文库,采用抗原固相淘选策略,通过三轮“结合-洗脱-扩增”的淘选流程完成噬菌体展示文库的富集,通过噬菌体ELISA鉴定阳性克隆,并进行抗体序列比对,以确定靶向TfR1的鲨鱼纳米抗体序列。构建鲨鱼纳米抗体重组表达载体,利用大肠杆菌原核表达系统进行鲨鱼纳米抗体的诱导表达,ELISA检测鲨鱼纳米抗体与TfR1重组蛋白的结合能力,竞争性ELISA检测鲨鱼纳米抗体与转铁蛋白(transferrin,Tf)竞争受体结合表位的情况,免疫荧光技术检测鲨鱼纳米抗体与脑微血管内皮细胞bEnd.3的结合能力,血脑屏障体外实验模型检测鲨鱼纳米抗体的跨血脑屏障转运能力。结果成功构建了库容量为2.35×10^(9) CFU的鲨鱼纳米抗体天然文库,该文库具备良好的多样性。经淘选获得2株靶向TfR1的鲨鱼纳米抗体,均具有亲和活性,其中D1具有跨血脑屏障转运功能。结论本研究成功筛选出靶向TfR1且具有跨血脑屏障转运功能的鲨鱼纳米抗体,为中枢神经系统疾病治疗提供了新的跨血脑屏障药物递送载体。
基金Research Projects of Health and Family Planning Commission of Sichuan Province.Project No:110530.
文摘Objective:To investigate the changes of Numb and TFR1 expression in ovarian cancer tissue and their clinical pathological value.Methods:90 patients with ovarian cancer underwent radical operation in Pidu District People's Hospital,the First Affiliated Hospital of Chengdu Medical College and Mianyang Central Hospital between August 2015 and April 2018,115 patients with ovarian chocolate cyst underwent surgery during the same period,and intraoperative lesion tissue samples of the two groups were collected and enrolled in ovarian cancer group and ovarian chocolate cyst group respectively.The expressions levels of Numb,TFR1 gene as well as ovarian cancer-related proliferation,invasion and autophagy genes in ovarian lesions were detected.Pearson test was used to evaluate the correlation of Numb and TFR1 gene expression in ovarian cancer tissues with proliferation,invasion and autophagy activity of cancer cells.Results:Numb mRNA expression in lesions of ovarian cancer group was lower than that of ovarian chocolate cyst group whereas TFR1 mRNA expression was higher than that of ovarian chocolate cyst group;proliferation genes C3G,Clusterin,FUNDC1,PIWIL4 and TNFAIP8 mRNA expression were higher than those of ovarian chocolate cyst group whereas Let-7i mRNA expression was lower than that of ovarian chocolate cyst group;invasion genes BRMS1,XAV939 and OVCA1 mRMA expression were lower than those of ovarian chocolate cyst group whereas CUL4A and MTA1 mRNA expression were higher than those of ovarian chocolate cyst group;autophagy genes FTY720 and Beclin1 mRNA expression were lower than those of ovarian chocolate cyst group whereas NAC-1 mRNA expression was higher than that of ovarian chocolate cyst group.Correlation analysis showed that the Numb and TFR1 gene expression in ovarian cancer tissues were directly correlated with the proliferation,invasion and autophagy activity of cancer cells.Conclusion:The abnormal decrease of Numb gene expression and the abnormal increase of TFR1 gene expression in ovarian cancer tissues are directly correlated with the malignant biological behaviors of cancer cells.
基金funded by the National Natural Science Foundation of China(grant Nos.82471912 and 82072436,China)Guangdong Basic and Applied Basic Research Foundation(2025A1515012449,China)the Provincial Key Laboratory of Immune Regulation and Immunotherapy(2022B1212010009,China).
文摘Heat stroke(HS)is a severe medical emergency characterized by coagulation and high mortality due to organ injury.This study identifies a novel mechanism in which platelet ferroptosis,driven by transferrin receptor 1(Tfr1)palmitoylation,significantly contributes to liver injury in HS.Our findings reveal a strong inverse correlation between platelet count and organ damage,especially liver injury,as well as mortality rates.Using murine models,we demonstrate that inhibiting Tfr1-mediated ferroptosis in platelets mitigates thrombocytopenia and decreases Interleukin-1β(IL-1β)secretion,thereby improving liver function and survival outcomes.This research highlights Tfr1 palmitoylation as a critical factor in iron transport within platelets,with the palmitoylation inhibitor 2-bromopalmitate(2BP)effectively reducing total iron,Fe2+,lipid ROS,4-hydroxynonenal(4-HNE),and cell cytotoxicity under heat stress.These results suggest that targeting Tfr1 palmitoylation-dependent ferroptosis in platelets offers a novel therapeutic strategy for treating HS-induced thrombocytopenia and liver injury.
基金supported by the National Key Research and Development Program of China(Grant No.2019YFC1604602-1)the Natural Science Foundation of Hebei Province(Grant No.19226631D).
文摘Mink enteritis virus(MEV),a member of the Protoparvovirus genus within the Parvovirinae subfamily,causes severe hemorrhagic enteritis and leukopenia in minks,thus representing a serious economic threat to mink farming.MEV,feline panleukopenia virus(FPV),and canine parvovirus(CPV)are closely related Protoparvovirus members;CPV and FPV infect hosts via binding to transferrin receptor 1(TfR1).However,the structural mechanism of MEV entry remains poorly understood.This study used cryo-electron microscopy(cryo-EM)and structural bioinformatics to determine the structure of MEV virus-like particles at 2.5 A resolution.The analysis revealed that MEV shares a highly conserved capsid architecture with CPV and FPV,particularly in theβ-barrel core and three-fold protrusions.We predict that MEV utilizes mink TfR1 for host entry,similar to CPV and FPV.Structural superposition and sequence alignment identified key residues in the MEV VP2 protein that are critical for mink TfR1 interaction.These findings provide structural and evolutionary insights into MEV host tropism,offering valuable information for efforts to mitigate the zoonotic risks of mammalian parvoviruses.
