1.Effect of cyclosporine A on autophagy-lysosomal pathway in tubular epithelial cells
Ning AN ; Zhihang LI ; Chen YANG ; Qingjun PAN ; Huafeng LIU
Chinese Journal of Nephrology 2017;33(4):284-289
Objective To investigate the effect of cyclosporine A (CsA) on autophagylysosomal pathway in tubular epithelial cells.Methods Human renal tubular epithelial cell line (HK-2 cell) was treated with different concentrations (3,5 and 10 μmol/L) of CsA for 24 h.Then the viability and apoptosis of cells were measured by MTT assay or AnnexinV-PI staining followed by flow cytometry analysis,respectively.Autophagy-related protein LC3-Ⅱ and p62 were detected by immunofluorescence assay.Autophagic flux was analyzed in HK-2 cells transfected with a tandem mRFP-GFP fluorescent-tagged LC3 (ffLC3) plasmid by laser confocal microscope.The lysosomal degradation was evaluated by DQ-ovalbumin staining followed by flow cytometry analysis.Results The viability of HK-2 cells was significantly decreased with CsA stimulation when compared with control group (P < 0.01),but the number of apoptotic cells was markedly increased by CsA treatment (P < 0.05).Compared with the control group,different doses of CsA dramatically increased the expressions of LC3-Ⅱ (P < 0.01) and p62 (P < 0.05) in HK-2 cells.Moreover,HK-2 cells treated with CsA displayed a significant increase in autophagosomes but a marked decrease in autolysosomes.In HK-2 cells,exposured to CsA caused a decrease in lysosomal degradation by DQ-ovalbumin staining when compared with control group (P < 0.01).Conclusion Blockade of autophagy via disrupting lysosome degradation may represent a novel mechanism of CsA-induced tubular epithelial cells injury.
2.The protective mechanism of TRPV4 channel inhibitor on blood-brain barrier damage after traumatic brain injury in rats
Fanhao KONG ; Hongyang ZHANG ; Wei ZHANG ; Mengze TANG ; Yingqiao WANG ; Xiang LI ; Xiaohui DING ; Zhihang YANG ; Hui XIE
Journal of Shenyang Medical College 2024;26(2):175-178
Objective:To investigate the protective mechanism of TRPV4 channel inhibitor on blood-brain barrier(BBB)damage after traumatic brain injury(TBI).Methods:The TBI rat model was established.TRPV4 channel inhibitor HC067047 or PKC-δ inhibitor Rottlerin was used to detect changes in BBB permeability,neurological function score,and the expression of microvascular endothelial tight junction proteins ZO-1 and ZO-2 in brain injury areas after TBI.Results:Compared with the Sham group,BBB permeability significantly increased,brain neurological function score significantly decreased,and the expression of ZO-1 and ZO-2 significantly decreased in TBI group(P<0.05).Compared with the TBI group,after administration of HC067047 or Rottlerin,changes in BBB permeability,brain neurological function score,the expression of ZO-1 and ZO-2 were partially reversed(P<0.05).Conclusions:TBI-induced BBB injury may be mediated by TRPV4 channel regulating PKC-δ signaling pathway to affect the expression of tight junction proteins ZO-1 and ZO-2.Inhibition of TRPV4 channel function or PKC-δ signal molecule can partially alleviate BBB damage induced by TBI.This study may provide new ideas for the treatment of clinical TBI.
3.Chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells induced by growth differentiation factor 5
Feifei LI ; Buyu WANG ; Zhihang YANG ; Xiaoyu DONG ; Jiang DENG
Chinese Journal of Tissue Engineering Research 2024;28(13):1976-1982
BACKGROUND:Growth differentiation factor 5 is a member of the transforming growth factor superfamily and one of the earliest markers of joint development.Growth differentiation factor 5 has an important role in cartilage repair. OBJECTIVE:To explore the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured.CCK-8 assay was used to detect the effect of different mass concentrations of growth differentiation factor 5 on the proliferation activity of bone marrow mesenchymal stem cells.RT-PCR was utilized to detect the expression of genes related to chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different mass concentrations of growth differentiation factor 5.To further investigate the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells,we added inhibitor XAV-939 and activator Laduviglusib of Wnt/β-catenin signaling pathways to induce cell culture for 14 days.RT-PCR and western blot assay were performed to detect the expression of cartilage-related genes and Wnt/β-catenin signaling pathway proteins. RESULTS AND CONCLUSION:(1)CCK-8 results showed no significant effect of growth differentiation factor 5 on the proliferation of bone marrow mesenchymal stem cells.(2)Growth differentiation factor 5 promoted the expression of cartilage-related genes type Ⅱ collagen,aggrecan and Sox9,among which growth differentiation factor 5 induced a significant upregulation of cartilage-related genes in the 50 ng/mL group.(3)Addition of Laduviglusib,an activator of Wnt/β-catenin signaling pathway,upregulated Sox9,β-catenin and type Ⅱ collagen expression(P<0.05).Addition of XAV939,an inhibitor of Wnt/β-catenin signaling pathway,down-regulated Sox9,β-catenin and type Ⅱ collagen expression(P<0.05).(4)Taken together,growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells may be associated with the activation of the Wnt/β-catenin signaling pathway.
4.Antiretroviral therapy-naïve people living with HIV tend to have more severe symptoms of COVID-19.
Jinfeng SUN ; Rui JIANG ; Yueming SHAO ; Jingjing HU ; Zhihang ZHENG ; Luling WU ; Li LIU ; Junyang YANG ; Yinzhong SHEN ; Renfang ZHANG ; Tangkai QI ; Jianjun SUN ; Zhenyan WANG ; Yang TANG ; Wei SONG ; Shuibao XU ; Bihe ZHAO ; Jun CHEN
Chinese Medical Journal 2023;136(22):2753-2755
5.Double-negative T cells: a promising avenue of adoptive cell therapy in transplant oncology.
Zhihang HU ; Modan YANG ; Hao CHEN ; Chiyu HE ; Zuyuan LIN ; Xinyu YANG ; Huigang LI ; Wei SHEN ; Di LU ; Xiao XU
Journal of Zhejiang University. Science. B 2023;24(5):387-396
Tumor recurrence is one of the major life-threatening complications after liver transplantation for liver cancer. In addition to the common mechanisms underlying tumor recurrence, another unavoidable problem is that the immunosuppressive therapeutic regimen after transplantation could promote tumor recurrence and metastasis. Transplant oncology is an emerging field that addresses oncological challenges in transplantation. In this context, a comprehensive therapeutic management approach is required to balance the anti-tumor treatment and immunosuppressive status of recipients. Double-negative T cells (DNTs) are a cluster of heterogeneous cells mainly consisting of two subsets stratified by T cell receptor (TCR) type. Among them, TCRαβ+ DNTs are considered to induce immune suppression in immune-mediated diseases, while TCRγδ+ DNTs are widely recognized as tumor killers. As a composite cell therapy, healthy donor-derived DNTs can be propagated to therapeutic numbers in vitro and applied for the treatment of several malignancies without impairing normal tissues or being rejected by the host. In this work, we summarized the biological characteristics and functions of DNTs in oncology, immunology, and transplantation. Based on the multiple roles of DNTs, we propose that a new balance could be achieved in liver transplant oncology using them as an off-the-shelf adoptive cell therapy (ACT).
Humans
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T-Lymphocytes
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Immunotherapy, Adoptive
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Neoplasm Recurrence, Local
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Transplantation, Homologous
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Cell- and Tissue-Based Therapy
6.GSH-responsive SN38 dimer-loaded shape-transformable nanoparticles with iRGD for enhancing chemo-photodynamic therapy.
Congcong LIN ; Fan TONG ; Rui LIU ; Rou XIE ; Ting LEI ; Yuxiu CHEN ; Zhihang YANG ; Huile GAO ; Xiangrong YU
Acta Pharmaceutica Sinica B 2020;10(12):2348-2361
Accurate tumor targeting, deep penetration and superb retention are still the main pursuit of developing excellent nanomedicine. To achieve these requirements, a stepwise stimuli-responsive strategy was developed through co-administration tumor penetration peptide iRGD with shape-transformable and GSH-responsive SN38-dimer (d-SN38)-loaded nanoparticles (d-SN38@NPs/iRGD). Upon intravenous injection, d-SN38@NPs with high drug loading efficiency (33.92 ± 1.33%) could effectively accumulate and penetrate into the deep region of tumor sites with the assistance of iRGD. The gathered nanoparticles simultaneously transformed into nanofibers upon 650 nm laser irradiation at tumor sites so as to promote their retention in the tumor and burst release of reactive oxygen species for photodynamic therapy. The loaded d-SN38 with disulfide bond responded to the high level of GSH in tumor cytoplasm, which consequently resulted in SN38 release and excellent chemo-photodynamic effect on tumor.
7.Erratum: Author correction to "A nanocleaner specifically penetrates the blood‒brain barrier at lesions to clean toxic proteins and regulate inflammation in Alzheimer's disease" Acta Pharmaceutica Sinica B 12, (2021) 4032-4044.
Ting LEI ; Zhihang YANG ; Xue XIA ; Yuxiu CHEN ; Xiaotong YANG ; Rou XIE ; Fan TONG ; Xiaolin WANG ; Huile GAO
Acta Pharmaceutica Sinica B 2022;12(6):2965-2967
[This corrects the article DOI: 10.1016/j.apsb.2021.04.022.].
8.A nanocleaner specifically penetrates the blood‒brain barrier at lesions to clean toxic proteins and regulate inflammation in Alzheimer's disease.
Ting LEI ; Zhihang YANG ; Xue XIA ; Yuxiu CHEN ; Xiaotong YANG ; Rou XIE ; Fan TONG ; Xiaolin WANG ; Huile GAO
Acta Pharmaceutica Sinica B 2021;11(12):4032-4044
Insurmountable blood‒brain barrier (BBB) and complex pathological features are the key factors affecting the treatment of Alzheimer's disease (AD). Poor accumulation of drugs in lesion sites and undesired effectiveness of simply reducing A