Biomedical subjects

Tong Qiao

Publications and source records attributed to Tong Qiao.

6 recordsLinked to original sources

PubMed2026

Impact of NR4A3 on wound healing in chronic venous ulcers and its association with the PI3K/Akt signaling pathway.

BACKGROUND: To investigate the role of NR4A3 in chronic venous ulcer (VU) wound healing and to explore its potential regulatory mechanism involving the PI3K/Akt pathway. METHODS: Differential expression and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the GSE174661 dataset. DEGs were filtered by |log2FC| > 1 and adjusted P < 0.05, with KEGG significance set at P < 0.05. NR4A3 was identified as the core gene. NR4A3 knockdown and overexpression were established in HaCaT cells to evaluate proliferation, migration, and inflammatory cytokines. TNF-α was used to mimic the inflammatory microenvironment. Western blotting assessed phosphorylation of GSK3β, mTOR, PI3K, and Akt. PI3K/Akt agonist 740Y-P and inhibitor LY294002 were used in rescue experiments. RESULTS: Bioinformatic analysis revealed that NR4A3 expression was markedly downregulated in chronic venous ulcer (VU) tissues relative to normal skin and ordinary acute wound tissues. Differentially expressed genes were significantly enriched in the PI3K/Akt signaling pathway. TNF-α stimulation significantly upregulated NR4A3 expression and increased phosphorylation of GSK3β and mTOR in HaCaT cells. In cultured HaCaT keratinocytes, NR4A3 knockdown suppressed cell proliferation and invasion, enhanced cell migration, and elevated the expression and secretion of pro-inflammatory cytokines (IL-6, IL-8, CXCL5), accompanied by reduced phosphorylation of PI3K and Akt. Conversely, NR4A3 overexpression promoted cell proliferation and invasion, restrained migration, and dampened inflammatory responses, while increasing PI3K/Akt phosphorylation. Treatment with the PI3K/Akt agonist 740Y-P partially rescued the impaired proliferation, aberrant migration, and excessive inflammation caused by NR4A3 silencing, whereas PI3K/Akt inhibitor LY294002 aggravated pathway suppression. These findings suggest that NR4A3-associated changes in keratinocyte functions and inflammatory reactions are functionally linked to PI3K/Akt pathway activity, and inflammatory stimulation activates GSK3β/mTOR signaling accompanied by compensatory NR4A3 upregulation. CONCLUSION: These findings suggest that NR4A3 is associated with keratinocyte behavior and inflammatory responses via the PI3K/Akt pathway, potentially affecting chronic VU progression and healing. Reduced NR4A3 may impair wound repair through inflammation and abnormal cell migration, while TNF-α induces compensatory NR4A3 elevation.

NR4A3↗

PubMed2006

Experimental study of recombinant eukaryotic expression vector of human eNOS in ECV304.

BACKGROUND AND PURPOSE: Gene transfer with recombinant non-viral vectors encoding vasodilator proteins, such as endothelial nitric oxide synthase (eNOS), maybe a preferential choice in gene therapy of artery restenosis following angioplasty, stent or anastomosis. However, the transfection rate of a non-viral vector, the harmful effects of eNOS transfection on endothelial cells (EC) and the control release of nitric oxide (NO) have been controversial. We designed the eukaryotic expression vector pcDNA3.1-eNOS to study the regulated expression of eNOS (in the presence of various chemical agents) and to evaluate the exogenous NO effect on EC proliferation in vitro. METHODS: The full-length human eNOS cDNA was inserted into the EcoRI cloning site of the pcDNA3.1 expression plasmid and the eNOS direction was tested by restriction enzyme digestion with XhoI to construct recombinant pcDNA3.1-eNOS. After co-transfection of pcDNA3.1-eNOS with pcDNA3.0-EGFP mediated by cationic liposomes into Human umbilical vein endothelial cells (ECV304), the transfection rate and the effect on ECV304 proliferation were calculated by fluorescence microscopy and flowcytometry. eNOS mRNA and protein were detected by reverse transcription-PCR (RT-PCR) and immunofluorescence, respectively. The eNOS activity, NO release and changes of the relevant cells growth curve were assessed after treating the transfected cells with four independent factors including ie Ca2+, L-arginine (L-Arg), Ethylene Diamine Tetraacetic Acid (EDTA) and N-nitro-L-arginine methylester (L-NAME). In addition, we examined the non-transfected cells status by isolated sodium nitroprusside (SNP) treatment. RESULT: eNOS cDNA was inserted into pcDNA3.1 in the proper direction. RT-PCR analysis showed that pcDNA3.1-eNOS transfected cells could express eNOS mRNA. The rate of eNOS transfection was 39.6 +/- 3.4%. Immunofluorescence staining displayed that subcellular localisation of eNOS was most prominent in plasma membrane and perinuclear regions of the cell. The eNOS activity of eNOS transfected cells had not increased significantly, whereas, in the presence of Ca2+, L-Arg, EDTA, and L-NAME, the eNOS activity was 96.98 +/- 13.47, 32.57 +/- 6.39, 11.63 +/- 3.02, 15.56 L 7.34 U/ml respectively and the NO level was 55.34 +/- 11.19, 9.43 +/- 4.51, 2.63 +/- 1.41, 3.73 +/- 1.65 mmol/L, respectively. Meanwhile, the growth curves of EC shifted. SNP also had obvious growth-inhibiting effects on the cells. Together, the ECV304 growth curve went downward in a NO concentration-dependent manner. CONCLUSION: Eukaryotic expression vector pcDNA3.1-eNOS was constructed successfully with the ability to express human eNOS mRNA and protein in EC effectively. The activity of eNOS in EC could be regulated by certain exogenous factors. Ca2+ was an important factor promoting NO release and excess NO had a cytotoxic effect on EC in vitro. Controlled release of NO in vivo and polygenic measurements might be considered in more clinical gene therapy studies.

Cell Line↗

PubMed2005

Interventional endovascular treatment for Budd-Chiari syndrome with long-term follow-up.

PURPOSE: To present results of a 9 year followup of percutaneous transluminal angioplasty (PTA) and stents placement in patients with Budd-Chiari Syndrome (BCS) and to evaluate the clinical value and applicability of this approach. MATERIALS AND METHODS: 44 consecutive patients with BCS (25 male and 19 female; average age, 42.6 years; age range, 19-77 years) were treated with PTA and stents during a 9-year period. The mean duration of symptoms was 46 months. Underlying active malignancy was the cause of occlusion in 5 patients. 3 patients had a history of taking oral contraceptives. The obstructed inferior vena cava (IVC) or hepatic vein (HV) were first dilated by a percutaneous transluminal balloon, and then a self-expanding stent was placed. Clinical patency was defined as absence or improvement of symptoms. Clinical follow-up was supplemented with colour Doppler sonography, CT scan, or both. RESULTS: Technical success was achieved in 97.2% (35/36) IVC and 83% (10/12) HV PTA and stents. Significantly, the IVC pressure decreased from 2.7 kPa (SD = 0.3) to 1.5 kPa (SD = 0.4) and HV pressure dropped from 2.3 kPa (SD = 0.4) to 1.3 kPa (SD = 0.2). The symptoms and signs disappeared or were relieved after operation in most of the patients. A few serious related complications including one stent migration and one pulmonary emboli occurred and were resolved in time. Patients were followed for a mean of 44 months (range 3-102). Short- and long-term results were satisfactory except for 3 patients (9.4%, 1 IVC, 2 HV) who presented with a restenosis or thrombosis and underwent additional therapy. There were 5 deaths owing to underlying malignant disease 3-17 months after the procedures. CONCLUSION: PTA and stent placements proved a safe and effective treatment in BCS and had a good long-term outcome and should be considered in patients who have symptoms or have no adequate alternative therapy.

Adult↗

PubMed2004

Inhibitory effect of antisense vascular endothelial growth factor 165 eukaryotic expression vector on proliferation of hepatocellular carcinoma cells.

AIM: To construct antisense VEGF(165) eukaryotic expression vector PCDNA(3)-as-VEGF(165) and to study its expression and effect on the proliferation of hepatocarcinoma SMMC-7721 cells. METHODS: VEGF(165) cDNA was inserted into polylinker sites of eukaryotic expression vector PCDNA(3) to construct PCDNA(3)-as-VEGF(165). Then the vector was transferred into human hepatocarcinoma cell strain SMMC-7721 with cation lipofectamine 2000 mediated methods to evaluate the expression of VEGF protein and the inhibitory effect on the proliferation of hepatocarcinoma SMMC-7721 cells. RESULTS: The detection indicated the presence of VEGF cDNA in normally cultured SMMC-7721 cells by PCR. VEGF mRNA expression was notably decreased in SMMC-7721 cells by RT-PCR after PCDNA(3)-as-VEGF(165) transfection. The expression of VEGF protein was dramatically inhibited (142.01+/-7.95 vs 1 625.52+/-64.46 pg/ml(-1), P<0.01) 2 days after transfection, which correlated with the dose of PCDNA(3)-as-VEGF(165)5 gene. VEGF protein was most expressed in PCDNA(3) transferred SMMC-7721 cells but few in PCDNA(3)-as-VEGF(165) transferred cells by immunohistochemical staining. The apoptotic rate of hepatocarcinoma SMMC-7721 cells was significantly promoted (17.98+/-0.86% vs 4.86+/-0.27%, P<0.01) and the survival rate was notably decreased (80.99+/-3.20% vs 93.52+/-3.93%, P<0.05) due to antisense VEGF(165) by flow cytometry (FCM). The transfection of antisense VEGF(165) gene resulted in the inhibitory effect on the proliferation of hepatocarcinoma cells by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and the death of all hepatocarcinoma cells on day 6 after transfection. CONCLUSION: It is confirmed that antisense VEGF(165) can inhibit the expression of VEGF protein, interfere with the proliferation and induce the apoptosis of hepatocarcinoma cells in our study. Antisense VEGF(165) gene therapy may play an important role in the treatment of human hepatocarcinoma.

Carcinoma, Hepatocellular↗

PubMed2004

Abdominal aorta transplantation after programmed cryopreservation.

AIM: To study the morphologic and cellular immunologic changes after homologous transplantation of the abdominal aorta in rats after programmed cryopreservation (-196 degrees). METHODS: Abdominal aorta was harvested from anesthetized Spraque Dawley (SD) rats for cryopreservation (group B) or immediate implantation (group A). The survival rates and apoptotic rates of aortic endothelial cells (ECs) were examined. The patency rates, histology and cellular immunologic changes of the abdominal aorta were examined on days 1, 3, 7, 14, 30, 60 after transplantation respectively. RESULTS: The survival rate of ECs after programmed cryopreservation was 90.1+/-1.79%, about 3.4% lower than that of uncryopreservation (93.5+/-1.96%). The apoptotic rates of ECs was increased after cryopreservation (7.15% vs 4.86%, P<0.05). The patency rate of group B was significantly higher than that of group A (91.6+/-12.9% vs 62.5+/-26.2%, P<0.01). CD4/CD8 ratio, TCR alpha beta and CD11b/CD18 ratio of group B were significantly lower than those of group A (P<0.05). Revivification of the cryopreserved abdominal aorta showed normal adventitia and intact smooth muscle cells. CONCLUSION: Cryopreservation can reduce homologous abdominal aortic antigenecity. Even if without administration of immunosuppressive agents, it is still feasible to implement homologous artery grafting in rats.

Animals↗

PubMed2003

Responses of vascular endothelial cells to angiogenic signaling are important for tumor cell survival.

Neoplastic cells overexpress several angiogenic cytokines, which stimulate neovascularization. Whether the responses of the host endothelial cells to these signaling molecules affect tumor cells during early tumorigenesis has not been investigated. We investigated pre-angiogenic tumor cell survival and angiogenesis initiation by two murine tumor lines (4T1 mammary carcinoma and B16 melanoma), which constitutively expressed GFP, in dorsal skin-fold window chambers of mice treated with extracellular domain of Tie-2 (ExTek) or bFGF. ExTek reduced tumor cell survival, retarded tumor growth, and inhibited angiogenesis onset compared with controls. bFGF increased tumor cell survival and promoted earlier angiogenesis and tumor growth. Neither bFGF nor ExTek affected cell proliferation in vitro. RT-PCR showed mRNA expression of bFGF receptor 2 (FGFR2) IIIb, which does not bind bFGF efficiently, by 4T1 cells and B16 cells express FGFR1 but not FGFR2. B16 cells expressed angiopoietin (Ang) 2, but neither cell line expresses Ang1. Both tumor lines express VEGF. These findings suggested that effects of bFGF and ExTek on tumor cell survival and angiogenesis were not due to direct action but were instead a result of paracrine factors secreted by endothelial cells. These subsequent signals from endothelial cells promote early survival and proliferation of disseminated tumor cells before onset of angiogenesis.

Angiogenesis Inducing Agents↗