The healing of diabetic wounds poses a significant healthcare burden due to persistent inflammation,M1 macrophage aggregation,and high glucose levels in the microenvironment.Previous studies have demonstrated that imm...The healing of diabetic wounds poses a significant healthcare burden due to persistent inflammation,M1 macrophage aggregation,and high glucose levels in the microenvironment.Previous studies have demonstrated that immunomodulatory hydrogel dressings can facilitate diabetic wound healing.However,current immunomodulatory hydrogels require costly and complex treatments such as cell therapy and cytokines.Herein,a hierarchical hydrogel dressing with continuous biochemical gradient based on glycyrrhizic acid(GA) was constructed to modulate immunomodulatory processes in diabetic wounds.The hydrogels present many desirable features,such as tunable mechanical properties,broad antibacterial ability,outstanding conductive,transparent,and self-adhesive properties.The resultant hydrogel can promote diabetic wound healing by preventing bacterial infection,promoting macrophage polarization,improving the inflammatory microenvironment,and inducing angiogenesis and neurogenesis.Furthermore,electrical stimulation(ES) can further promote the healing of chronic diabetic wounds,providing valuable guidance for relevant clinical practice.展开更多
Intervertebral disc herniation(IVDH)is a common manifestation of intervertebral disc degeneration(IVDD)characterized by inflammation that results in the rupture of the annulus fibrosus(AF)and her-niation of the nucleu...Intervertebral disc herniation(IVDH)is a common manifestation of intervertebral disc degeneration(IVDD)characterized by inflammation that results in the rupture of the annulus fibrosus(AF)and her-niation of the nucleus pulposus(NP).While current clinical research primarily focuses on regulating the degenerative NP,the crucial role of the AF in maintaining the mechanical stability and metabolic balance of the intervertebral disc(IVD)has been overlooked.Resolving immunoregulation and AF repair is im-perative to effectively prevent recurrent herniation.Therefore,this study introduces a bioactive sealant(OD/GM/QCS-sEVs),which combines gelatin methacryloyl(GM)and oxidized dextran(OD)with quater-nized chitosan(QCS)and incorporates small extracellular vesicles(sEVs).The developed sealant possesses injectability,self-healing capabilities,tissue adhesiveness,and mechanical stability,with an average ad-hesive strength of 109.63 kPa.In vitro experiments demonstrate that OD/GM/QCS-sEVs effectively seal AF defects while preserving mechanical properties comparable to those of a normal IVD.Additionally,the sealant releases sEVs through a pH-responsive mechanism,thereby modulating macrophage polarization to the M2 phenotype via the NF-κB signaling pathway.This mechanism facilitates immunoregulation and anti-inflammatory effects,and promotes stem cell differentiation into fibrocartilage.Animal experiments confirm the ability of OD/GM/QCS-sEVs to seal defects,prevent proteoglycan loss,inhibit IVDD develop-ment,and promote AF regeneration.Overall,OD/GM/QCS-sEVs hold promise as an innovative bioactive sealant for recurrent herniation by resolving immunoregulation and AF regeneration.展开更多
【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少...【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少PCV2感染造成的经济损失。【方法】利用实验室构建的重组表达宿主菌pET32a-PD1/Rosetta(DE)进行诱导表达、纯化,获得猪可溶性PD-1蛋白(soluble PD-1,sPD-1)的诱导表达条件;制备大量具有活性的猪sPD-1蛋白,体外作用于PCV2感染的猪外周血单个核细胞(peripheral blood mononuclear cell,PBMCs),分别通过CCK-8和流式细胞术检测PBMCs的增殖情况,间接免疫荧光法和RT-qPCR检测猪sPD-1蛋白对PCV2病毒载量的影响,RT-qPCR和ELISA检测猪sPD-1蛋白对免疫相关细胞因子IL-2、IL-12、IL-21、IL-17A和IFN-γ等的转录水平和分泌水平的影响。【结果】通过诱导表达、纯化获得了高纯度、高活性的重组猪sPD-1蛋白。与未使用猪sPD-1蛋白处理的PCV2感染PBMCs组相比,10μg·mL−1猪sPD-1蛋白处理的猪PCV2病毒载量下降至1000 copies·μL−1以下;CCK-8检测结果显示猪sPD-1蛋白处理组细胞增殖指数显著提高(P<0.05);流式细胞术检测发现猪sPD-1蛋白组的平均荧光强度为(68.60±10.14)%,相对于PCV2病毒组(28.70±3.18)%显著增强(P<0.05);反转录荧光定量PCR和ELISA检测,发现猪sPD-1蛋白处理组的细胞因子IL-2、IL-12和IFN-γ的转录水平和分泌水平显著升高(P<0.05)。【结论】猪sPD-1蛋白可以通过阻断PD-1/PD-Ls通路降低PCV2的病毒载量,促进PBMCs的增殖,增强PBMCs细胞因子的转录水平和分泌水平免疫反应。因此,猪sPD-1蛋白在体外可以增强PCV2感染PBMCs的免疫反应,进一步为病毒性疾病的防控提供理论依据。展开更多
本研究旨在探讨鲟鱼子酱对环磷酰胺(Cyclophosphamide,CTX)诱导的免疫低下小鼠的免疫增强作用。以ICR(Institute of Cancer Research,ICR)小鼠为试验对象,腹腔注射CTX建立免疫低下模型,随后对免疫低下小鼠投喂添加低(4.2 g/kg)、中(8.4 ...本研究旨在探讨鲟鱼子酱对环磷酰胺(Cyclophosphamide,CTX)诱导的免疫低下小鼠的免疫增强作用。以ICR(Institute of Cancer Research,ICR)小鼠为试验对象,腹腔注射CTX建立免疫低下模型,随后对免疫低下小鼠投喂添加低(4.2 g/kg)、中(8.4 g/kg)、高(16.8 g/kg)剂量杂交鲟鱼子酱的鼠粮,对照组投喂不添加鱼子酱的鼠粮,为期30 d。通过H&E染色观察小鼠肠组织形态,并检测小鼠免疫器官指数、免疫细胞活力、免疫酶活和免疫因子表达水平。结果表明,与模型组相比,鱼子酱添加高剂量组能提升小鼠日采食量、体重,缓解小鼠肠绒毛损伤;高剂量鱼子酱将小鼠脾脏和胸腺器官指数、巨噬细胞吞噬能力和增殖活性显著提高了31.1%、40.1%和25.2%、48.5%(P<0.05),高剂量鱼子酱将小鼠T、B淋巴细胞增殖活性极显著提高了32.9%、50.9%(P<0.01);中剂量鱼子酱将小鼠乳酸脱氢酶、酸性磷酸酶活力显著增加了33%、13.4%(P<0.05),高剂量鱼子酱将小鼠溶菌酶含量极显著提高了53.4%(P<0.01);高剂量鱼子酱显著提升了小鼠脾脏免疫细胞因子白介素-2(Interleukin-2,IL-2)、白介素-12(Interleukin-12,IL-12)和肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)的mRNA表达水平,分别提升了86.9%、86.1%和91.5%(P<0.05),低剂量鱼子酱将白介素-6(Interleukin-6,IL-6)的mRNA表达水平显著提高了64.5%(P<0.05);此外,高剂量鱼子酱显著促进小鼠血清中IL-2、IL-12和TNF-α的分泌(P<0.05),分别上升了52.1%、30.6%和5%,中剂量鱼子酱将小鼠IL-6的分泌极显著提升了57.8%(P<0.01)。综上,鱼子酱能够减轻CTX诱导的小鼠肠道损伤,有利于肠绒毛的恢复,增强了CTX抑制的细胞免疫、体液免疫和非特异性免疫,从而增强机体的免疫调节能力。展开更多
基金supported by Natural Science Foundation of Jilin Province(No.SKL202302002)。
文摘The healing of diabetic wounds poses a significant healthcare burden due to persistent inflammation,M1 macrophage aggregation,and high glucose levels in the microenvironment.Previous studies have demonstrated that immunomodulatory hydrogel dressings can facilitate diabetic wound healing.However,current immunomodulatory hydrogels require costly and complex treatments such as cell therapy and cytokines.Herein,a hierarchical hydrogel dressing with continuous biochemical gradient based on glycyrrhizic acid(GA) was constructed to modulate immunomodulatory processes in diabetic wounds.The hydrogels present many desirable features,such as tunable mechanical properties,broad antibacterial ability,outstanding conductive,transparent,and self-adhesive properties.The resultant hydrogel can promote diabetic wound healing by preventing bacterial infection,promoting macrophage polarization,improving the inflammatory microenvironment,and inducing angiogenesis and neurogenesis.Furthermore,electrical stimulation(ES) can further promote the healing of chronic diabetic wounds,providing valuable guidance for relevant clinical practice.
基金supported by the National Natural Science Foundation of China(Grant Nos.51873069,52272276,52073103,52203164)the Zhongshan Innovation Project of high-end Scientific Research Institutions(Grant No.2020AG020)+2 种基金the Key-Area Research and Development Program of Guangdong Province(No.2020B090924004)Beijing Municipal Health Commission(Grant Nos.BMHC-2018-4,BMHC-2019-9,PXM2020026275000002)the Postdoctoral Research Foundation of China(No.2022M711183).
文摘Intervertebral disc herniation(IVDH)is a common manifestation of intervertebral disc degeneration(IVDD)characterized by inflammation that results in the rupture of the annulus fibrosus(AF)and her-niation of the nucleus pulposus(NP).While current clinical research primarily focuses on regulating the degenerative NP,the crucial role of the AF in maintaining the mechanical stability and metabolic balance of the intervertebral disc(IVD)has been overlooked.Resolving immunoregulation and AF repair is im-perative to effectively prevent recurrent herniation.Therefore,this study introduces a bioactive sealant(OD/GM/QCS-sEVs),which combines gelatin methacryloyl(GM)and oxidized dextran(OD)with quater-nized chitosan(QCS)and incorporates small extracellular vesicles(sEVs).The developed sealant possesses injectability,self-healing capabilities,tissue adhesiveness,and mechanical stability,with an average ad-hesive strength of 109.63 kPa.In vitro experiments demonstrate that OD/GM/QCS-sEVs effectively seal AF defects while preserving mechanical properties comparable to those of a normal IVD.Additionally,the sealant releases sEVs through a pH-responsive mechanism,thereby modulating macrophage polarization to the M2 phenotype via the NF-κB signaling pathway.This mechanism facilitates immunoregulation and anti-inflammatory effects,and promotes stem cell differentiation into fibrocartilage.Animal experiments confirm the ability of OD/GM/QCS-sEVs to seal defects,prevent proteoglycan loss,inhibit IVDD develop-ment,and promote AF regeneration.Overall,OD/GM/QCS-sEVs hold promise as an innovative bioactive sealant for recurrent herniation by resolving immunoregulation and AF regeneration.
文摘【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少PCV2感染造成的经济损失。【方法】利用实验室构建的重组表达宿主菌pET32a-PD1/Rosetta(DE)进行诱导表达、纯化,获得猪可溶性PD-1蛋白(soluble PD-1,sPD-1)的诱导表达条件;制备大量具有活性的猪sPD-1蛋白,体外作用于PCV2感染的猪外周血单个核细胞(peripheral blood mononuclear cell,PBMCs),分别通过CCK-8和流式细胞术检测PBMCs的增殖情况,间接免疫荧光法和RT-qPCR检测猪sPD-1蛋白对PCV2病毒载量的影响,RT-qPCR和ELISA检测猪sPD-1蛋白对免疫相关细胞因子IL-2、IL-12、IL-21、IL-17A和IFN-γ等的转录水平和分泌水平的影响。【结果】通过诱导表达、纯化获得了高纯度、高活性的重组猪sPD-1蛋白。与未使用猪sPD-1蛋白处理的PCV2感染PBMCs组相比,10μg·mL−1猪sPD-1蛋白处理的猪PCV2病毒载量下降至1000 copies·μL−1以下;CCK-8检测结果显示猪sPD-1蛋白处理组细胞增殖指数显著提高(P<0.05);流式细胞术检测发现猪sPD-1蛋白组的平均荧光强度为(68.60±10.14)%,相对于PCV2病毒组(28.70±3.18)%显著增强(P<0.05);反转录荧光定量PCR和ELISA检测,发现猪sPD-1蛋白处理组的细胞因子IL-2、IL-12和IFN-γ的转录水平和分泌水平显著升高(P<0.05)。【结论】猪sPD-1蛋白可以通过阻断PD-1/PD-Ls通路降低PCV2的病毒载量,促进PBMCs的增殖,增强PBMCs细胞因子的转录水平和分泌水平免疫反应。因此,猪sPD-1蛋白在体外可以增强PCV2感染PBMCs的免疫反应,进一步为病毒性疾病的防控提供理论依据。
文摘本研究旨在探讨鲟鱼子酱对环磷酰胺(Cyclophosphamide,CTX)诱导的免疫低下小鼠的免疫增强作用。以ICR(Institute of Cancer Research,ICR)小鼠为试验对象,腹腔注射CTX建立免疫低下模型,随后对免疫低下小鼠投喂添加低(4.2 g/kg)、中(8.4 g/kg)、高(16.8 g/kg)剂量杂交鲟鱼子酱的鼠粮,对照组投喂不添加鱼子酱的鼠粮,为期30 d。通过H&E染色观察小鼠肠组织形态,并检测小鼠免疫器官指数、免疫细胞活力、免疫酶活和免疫因子表达水平。结果表明,与模型组相比,鱼子酱添加高剂量组能提升小鼠日采食量、体重,缓解小鼠肠绒毛损伤;高剂量鱼子酱将小鼠脾脏和胸腺器官指数、巨噬细胞吞噬能力和增殖活性显著提高了31.1%、40.1%和25.2%、48.5%(P<0.05),高剂量鱼子酱将小鼠T、B淋巴细胞增殖活性极显著提高了32.9%、50.9%(P<0.01);中剂量鱼子酱将小鼠乳酸脱氢酶、酸性磷酸酶活力显著增加了33%、13.4%(P<0.05),高剂量鱼子酱将小鼠溶菌酶含量极显著提高了53.4%(P<0.01);高剂量鱼子酱显著提升了小鼠脾脏免疫细胞因子白介素-2(Interleukin-2,IL-2)、白介素-12(Interleukin-12,IL-12)和肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)的mRNA表达水平,分别提升了86.9%、86.1%和91.5%(P<0.05),低剂量鱼子酱将白介素-6(Interleukin-6,IL-6)的mRNA表达水平显著提高了64.5%(P<0.05);此外,高剂量鱼子酱显著促进小鼠血清中IL-2、IL-12和TNF-α的分泌(P<0.05),分别上升了52.1%、30.6%和5%,中剂量鱼子酱将小鼠IL-6的分泌极显著提升了57.8%(P<0.01)。综上,鱼子酱能够减轻CTX诱导的小鼠肠道损伤,有利于肠绒毛的恢复,增强了CTX抑制的细胞免疫、体液免疫和非特异性免疫,从而增强机体的免疫调节能力。
