Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis...Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis.However,the regulatory impact of TRPC6 on renal fibrosis remains unclear.Methods In vivo,TRPC6 knockout(TRPC6−/−)mice and age-matched 129 SvEv(WT)mice underwent unilateral renal ischemia-reperfusion(uIR)injury surgery on the left renal pedicle or sham operation.Kidneys and serum were collected on days 7,14,21,and 28 after euthanasia.In vitro,primary tubular epithelial cells(PTECs)were isolated from TRPC6−/−and WT mice,followed by treatment with transforming growth factorβ1(TGFβ1)for 72 h.The anti-fibrotic effect of TRPC6−/−and the underlying mechanisms were assessed through hematoxylin-eosin staining,Masson staining,immunostaining,qRT-PCR,and Western blotting.Results Increased TRPC6 expression was observed in uIR mice and PTECs treated with TGFβ1.TRPC6−/−alleviated renal fibrosis by reducing the expression of fibrotic markers(Col-1,α-SMA,and vimentin),as well as decreasing the apoptosis and inflammation of PTECs during fibrotic progression both in vivo and in vitro.Additionally,we found that the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/glycogen synthase kinase 3 beta(GSK3β)signaling pathway,a pivotal player in renal fibrosis,was down-regulated following TRPC6 deletion.Conclusion These results suggest that the ablation of TRPC6 may mitigate renal fibrosis by inhibiting the apoptosis and inflammation of PTECs through down-regulation of the PI3K/AKT/GSK3βpathway.Targeting TRPC6 could be a novel therapeutic strategy for preventing chronic kidney disease.展开更多
AIM:To investigate whether nintedanib can inhibit pterygium cells through the fibroblast growth factor receptor 2(FGFR2)/extracellular-signal-regulated kinase(ERK)pathway.METHODS:Human primary pterygium cells were cul...AIM:To investigate whether nintedanib can inhibit pterygium cells through the fibroblast growth factor receptor 2(FGFR2)/extracellular-signal-regulated kinase(ERK)pathway.METHODS:Human primary pterygium cells were cultured in vitro.After treatment with nintedanib,the cell morphology was observed under microscopy,the morphological changes of the nucleus were observed after DAPI staining,apoptosis was analyzed by Annexin-V FITC/PI double staining,and the changes of apoptosis-associated proteins were detected by Western blot.The binding ability of nintedanib to FGFR2 was predicted by molecular docking.Finally,by silencing FGFR2,we explored whether nintedanib inhibited FGFR2/ERK pathway.RESULTS:The results showed that nintedanib inhibited the growth of pterygium cells and caused nuclear pyknosis.The results of Annexin-VFITC/PI double staining showed that nintedanib was able to induce early and late apoptosis of pterygium cells,significantly increasing the expression of apoptosis-associated proteins Bax and cleaved-Caspase3(P<0.05),and reducing the expression of Bcl-2(P<0.05).In addition,nintedanib significantly inhibited ERK1/2 phosphorylation through FGFR2(P<0.05).After silencing the expression of FGFR2,there was no significant difference in the inhibition of ERK1/2 phosphorylation by nintedanib(P>0.05).CONCLUSION:Nintedanib induces apoptosis of pterygium cells by inhibiting FGFR2/ERK pathway.展开更多
AIM:To investigate the effects of nintedanib thermo-sensitive hydrogel(NTH)on neovascularization and related markers in corneal alkali burns of Wistar rats.METHODS:NTH was prepared by grinding,and its phase-transition...AIM:To investigate the effects of nintedanib thermo-sensitive hydrogel(NTH)on neovascularization and related markers in corneal alkali burns of Wistar rats.METHODS:NTH was prepared by grinding,and its phase-transition temperature was determined.Thirty specific-pathogen-free Wistar rats served as a model of corneal alkali burn in the right eye were randomly divided into 3 groups(n=10,each):model group treated with 0.9%saline once a day,NTH group with 0.2%nintedanib b.i.d,and dexamethasone group with dexamethasone ointment once a day.The left eye of rats served as the controls.The corneal transparency was observed under a slit-lamp microscope,and the area of neovascularization was calculated.On day 7,the rats were sacrificed,and the cornea was removed and embedded with paraffin,then stained with hematoxylin一eosin,and the expression of vascular endothelial growth factor receptor 2(VEGFR-2)and CD31 in the corneal tissues of each group was detected by immunofluorescence.RESULTS:The phase-transition temperature ofnintedanib obtained by grinding was 37℃after adding artificial tears.The results of the alkali burn model indicated that the growth rate of neovascularization in the NTH group was slower than that in the model group,and the neovascularization area was significantly smaller than that in the model group(P<0.05).Moreover,CD31 and VEGFR-2 expression levels in the NTH group were significantly lower than those in the model group.CONCLUSION:NTH becomes colloidal at body temperature,which is beneficial for releasing the drug slowly and can significantly inhibit the neovascularization of corneal induced by alkali burn in rats.展开更多
AIM:To introduce and evaluate the clinical efficacy of a new technique,the use of viscoelastic substances(VS)to close leaking sclerotomy in 23G microincision vitrectomy,and to observe its effect on the visual acuity a...AIM:To introduce and evaluate the clinical efficacy of a new technique,the use of viscoelastic substances(VS)to close leaking sclerotomy in 23G microincision vitrectomy,and to observe its effect on the visual acuity and intraocular pressure(IOP)of patients.METHODS:Patients who underwent 23G vitrectomy in Ningbo Eye Hospital before the use of VS technique(June 2019 to September 2020)and after the use of VS technique(October 2020 to December 2021)were selected as the subjects of this study.The above cases underwent operation by the same surgeon and were retrospectively analyzed.VS technique was used as the alternative to suturing,in which a small amount of VS was injected at the leaking sclerotomy and then gently massaged to confirm leaking sclerotomy closure.RESULTS:A total of 174 eyes were covered in the study,including 84 eyes in the control group(before the use of VS technique)and 90 eyes in the VS technique group.The number of eyes that needed to be sutured decreased considerably from 42.9%in the control group to 3.3%in the VS technique group,and the proportion of subconjunctival hemorrhage at 1-2d after surgery decreased remarkably from 35.7%in the control group to 2.2%in the VS technique group.No substantial differences in the incidence of mean IOP and low IOP were found between 1-2 and 3-20d after surgery in the VS technique group.No major complications associated with VS technique were identified during the study.CONCLUSION:In 23G microincision vitrectomy,VS technique is a safe,simple,and effective method to close leaking sclerotomy.展开更多
基金supported by grants from the National Natural Science Foundation of China(Nos.31171087,30970662,and 82100715)the Henan Medical Science and Technology Research Program(Joint Construction,No.LHGJ20190246).
文摘Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis.However,the regulatory impact of TRPC6 on renal fibrosis remains unclear.Methods In vivo,TRPC6 knockout(TRPC6−/−)mice and age-matched 129 SvEv(WT)mice underwent unilateral renal ischemia-reperfusion(uIR)injury surgery on the left renal pedicle or sham operation.Kidneys and serum were collected on days 7,14,21,and 28 after euthanasia.In vitro,primary tubular epithelial cells(PTECs)were isolated from TRPC6−/−and WT mice,followed by treatment with transforming growth factorβ1(TGFβ1)for 72 h.The anti-fibrotic effect of TRPC6−/−and the underlying mechanisms were assessed through hematoxylin-eosin staining,Masson staining,immunostaining,qRT-PCR,and Western blotting.Results Increased TRPC6 expression was observed in uIR mice and PTECs treated with TGFβ1.TRPC6−/−alleviated renal fibrosis by reducing the expression of fibrotic markers(Col-1,α-SMA,and vimentin),as well as decreasing the apoptosis and inflammation of PTECs during fibrotic progression both in vivo and in vitro.Additionally,we found that the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/glycogen synthase kinase 3 beta(GSK3β)signaling pathway,a pivotal player in renal fibrosis,was down-regulated following TRPC6 deletion.Conclusion These results suggest that the ablation of TRPC6 may mitigate renal fibrosis by inhibiting the apoptosis and inflammation of PTECs through down-regulation of the PI3K/AKT/GSK3βpathway.Targeting TRPC6 could be a novel therapeutic strategy for preventing chronic kidney disease.
文摘AIM:To investigate whether nintedanib can inhibit pterygium cells through the fibroblast growth factor receptor 2(FGFR2)/extracellular-signal-regulated kinase(ERK)pathway.METHODS:Human primary pterygium cells were cultured in vitro.After treatment with nintedanib,the cell morphology was observed under microscopy,the morphological changes of the nucleus were observed after DAPI staining,apoptosis was analyzed by Annexin-V FITC/PI double staining,and the changes of apoptosis-associated proteins were detected by Western blot.The binding ability of nintedanib to FGFR2 was predicted by molecular docking.Finally,by silencing FGFR2,we explored whether nintedanib inhibited FGFR2/ERK pathway.RESULTS:The results showed that nintedanib inhibited the growth of pterygium cells and caused nuclear pyknosis.The results of Annexin-VFITC/PI double staining showed that nintedanib was able to induce early and late apoptosis of pterygium cells,significantly increasing the expression of apoptosis-associated proteins Bax and cleaved-Caspase3(P<0.05),and reducing the expression of Bcl-2(P<0.05).In addition,nintedanib significantly inhibited ERK1/2 phosphorylation through FGFR2(P<0.05).After silencing the expression of FGFR2,there was no significant difference in the inhibition of ERK1/2 phosphorylation by nintedanib(P>0.05).CONCLUSION:Nintedanib induces apoptosis of pterygium cells by inhibiting FGFR2/ERK pathway.
基金Supported by Zhejiang Province Traditional Chinese Medicine Science and Technology Plan Project(No.2018ZA111)Natural Science Foundation of Zhejiang Province(No.LY19H120001)+4 种基金Zhejiang Provincial Medicine and Health Science and Technology Project(No.2020KY288)Ningbo Natural Science Foundation(No.2019A610353)Ningbo Leading and Outstanding Talents Cultivation Project Selects and Supports Scientific Research Projects(No.NBLJ201801037)Ningbo Public Welfare Science and Technology Plan Project(No.2019C50059)Ningbo Yinzhou District Science and Technology Bureau Agricultural and Social Science and Technology Plan Project(Yanke[2017]No.110)。
文摘AIM:To investigate the effects of nintedanib thermo-sensitive hydrogel(NTH)on neovascularization and related markers in corneal alkali burns of Wistar rats.METHODS:NTH was prepared by grinding,and its phase-transition temperature was determined.Thirty specific-pathogen-free Wistar rats served as a model of corneal alkali burn in the right eye were randomly divided into 3 groups(n=10,each):model group treated with 0.9%saline once a day,NTH group with 0.2%nintedanib b.i.d,and dexamethasone group with dexamethasone ointment once a day.The left eye of rats served as the controls.The corneal transparency was observed under a slit-lamp microscope,and the area of neovascularization was calculated.On day 7,the rats were sacrificed,and the cornea was removed and embedded with paraffin,then stained with hematoxylin一eosin,and the expression of vascular endothelial growth factor receptor 2(VEGFR-2)and CD31 in the corneal tissues of each group was detected by immunofluorescence.RESULTS:The phase-transition temperature ofnintedanib obtained by grinding was 37℃after adding artificial tears.The results of the alkali burn model indicated that the growth rate of neovascularization in the NTH group was slower than that in the model group,and the neovascularization area was significantly smaller than that in the model group(P<0.05).Moreover,CD31 and VEGFR-2 expression levels in the NTH group were significantly lower than those in the model group.CONCLUSION:NTH becomes colloidal at body temperature,which is beneficial for releasing the drug slowly and can significantly inhibit the neovascularization of corneal induced by alkali burn in rats.
基金Supported by General Class A of the Zhejiang Medical and Health Science and Technique Plan Project in 2020(No.2020ky288)Zhejiang Provincial Science and Technique Program of Traditional Chinese Medicine(No.2021ZB268)+2 种基金Health Science and Technique Program of Zhejiang Province(No.2021PY073)Yinzhou District Agriculture and Social Development Field Science and Technique Project(No.2021AS0058No.2020AS0080)。
文摘AIM:To introduce and evaluate the clinical efficacy of a new technique,the use of viscoelastic substances(VS)to close leaking sclerotomy in 23G microincision vitrectomy,and to observe its effect on the visual acuity and intraocular pressure(IOP)of patients.METHODS:Patients who underwent 23G vitrectomy in Ningbo Eye Hospital before the use of VS technique(June 2019 to September 2020)and after the use of VS technique(October 2020 to December 2021)were selected as the subjects of this study.The above cases underwent operation by the same surgeon and were retrospectively analyzed.VS technique was used as the alternative to suturing,in which a small amount of VS was injected at the leaking sclerotomy and then gently massaged to confirm leaking sclerotomy closure.RESULTS:A total of 174 eyes were covered in the study,including 84 eyes in the control group(before the use of VS technique)and 90 eyes in the VS technique group.The number of eyes that needed to be sutured decreased considerably from 42.9%in the control group to 3.3%in the VS technique group,and the proportion of subconjunctival hemorrhage at 1-2d after surgery decreased remarkably from 35.7%in the control group to 2.2%in the VS technique group.No substantial differences in the incidence of mean IOP and low IOP were found between 1-2 and 3-20d after surgery in the VS technique group.No major complications associated with VS technique were identified during the study.CONCLUSION:In 23G microincision vitrectomy,VS technique is a safe,simple,and effective method to close leaking sclerotomy.
