1.Inhibitory effects of different concentrations of auranofin on M1 macrophage function and its therapeutic potential in diabetic wound healing
Hongfei PAN ; Zhenbing ZHUANG ; Baiyun XU ; Zhangyang YANG ; Kairui LIN ; Bingqing ZHAN ; Jinghan LAN ; Heng GAO ; Nanbo ZHANG ; Jiayu LIN
Chinese Journal of Tissue Engineering Research 2026;30(6):1390-1397
BACKGROUND:During diabetic wound healing,the sustained activation of M1 macrophages exacerbates the inflammatory response and hinders wound repair.Auranofin,an anti-inflammatory drug,has not been clearly studied for its effects on M1 macrophages and its potential role in diabetic wound healing.OBJECTIVE:To investigate the effects of different concentrations of auranofin on the biological function of M1 macrophages and evaluate its potential application in diabetic wound healing.METHODS:RAW264.7 and THP-1 cells were used as research models.M1 polarization was induced using different concentrations of interferon-γ and lipopolysaccharide.M1 macrophages were treated with 1 and 2 μmol/L auranofin.Cell counting kit-8 assay was used to evaluate the effect of auranofin on cell viability.Quantitative real-time PCR was performed to detect mRNA expression of interleukin-1β,interleukin-6,and tumor necrosis factor-α.ELISA was employed to measure the levels of interleukin-1β,interleukin-6,and tumor necrosis factor-α in the supernatant.Western blot analysis was used to assess the expression of nuclear factor-κB(p65),phosphorylated mitogen-activated protein kinases(MAPK),and total MAPK proteins.Additionally,6-8-week-old male C57BL/6J and db/db diabetic mice were used for wound healing experiments,with the mice divided into C57 control,db/db control and auranofin treatment groups,each containing six animals.Dorsal skin defect modeling and treatment with intraperitoneal injection of auranofin were performed to observe wound healing in mice.RESULTS AND CONCLUSION:(1)Cell experiments showed that co-treatment with interferon-y(10 ng/mL)and lipopolysaccharide(100 ng/mL)significantly induced M1 polarization in RAW264.7 and THP-1 cells,resulting in increased mRNA expression of interleukin-1β,interleukin-6,and tumor necrosis factor-α.Treatment with auranofin(1 and 2 μmol/L)reduced the mRNA expression of these inflammatory factors in the cells and inhibited the secretion of inflammatory factors in the cell supernatant.(2)Auranofin treatment significantly suppressed the activation of nuclear factor-κB(p65)and phosphorylated MAPK signaling pathways.(3)Animal experiments showed that auranofin promoted wound healing in db/db diabetic mice,suggesting that auranofin has strong anti-inflammatory effects and may facilitate the healing of wounds in diabetic mice.
2.Inhibitory effects of different concentrations of auranofin on M1 macrophage function and its therapeutic potential in diabetic wound healing
Hongfei PAN ; Zhenbing ZHUANG ; Baiyun XU ; Zhangyang YANG ; Kairui LIN ; Bingqing ZHAN ; Jinghan LAN ; Heng GAO ; Nanbo ZHANG ; Jiayu LIN
Chinese Journal of Tissue Engineering Research 2026;30(6):1390-1397
BACKGROUND:During diabetic wound healing,the sustained activation of M1 macrophages exacerbates the inflammatory response and hinders wound repair.Auranofin,an anti-inflammatory drug,has not been clearly studied for its effects on M1 macrophages and its potential role in diabetic wound healing.OBJECTIVE:To investigate the effects of different concentrations of auranofin on the biological function of M1 macrophages and evaluate its potential application in diabetic wound healing.METHODS:RAW264.7 and THP-1 cells were used as research models.M1 polarization was induced using different concentrations of interferon-γ and lipopolysaccharide.M1 macrophages were treated with 1 and 2 μmol/L auranofin.Cell counting kit-8 assay was used to evaluate the effect of auranofin on cell viability.Quantitative real-time PCR was performed to detect mRNA expression of interleukin-1β,interleukin-6,and tumor necrosis factor-α.ELISA was employed to measure the levels of interleukin-1β,interleukin-6,and tumor necrosis factor-α in the supernatant.Western blot analysis was used to assess the expression of nuclear factor-κB(p65),phosphorylated mitogen-activated protein kinases(MAPK),and total MAPK proteins.Additionally,6-8-week-old male C57BL/6J and db/db diabetic mice were used for wound healing experiments,with the mice divided into C57 control,db/db control and auranofin treatment groups,each containing six animals.Dorsal skin defect modeling and treatment with intraperitoneal injection of auranofin were performed to observe wound healing in mice.RESULTS AND CONCLUSION:(1)Cell experiments showed that co-treatment with interferon-y(10 ng/mL)and lipopolysaccharide(100 ng/mL)significantly induced M1 polarization in RAW264.7 and THP-1 cells,resulting in increased mRNA expression of interleukin-1β,interleukin-6,and tumor necrosis factor-α.Treatment with auranofin(1 and 2 μmol/L)reduced the mRNA expression of these inflammatory factors in the cells and inhibited the secretion of inflammatory factors in the cell supernatant.(2)Auranofin treatment significantly suppressed the activation of nuclear factor-κB(p65)and phosphorylated MAPK signaling pathways.(3)Animal experiments showed that auranofin promoted wound healing in db/db diabetic mice,suggesting that auranofin has strong anti-inflammatory effects and may facilitate the healing of wounds in diabetic mice.
3.PD-1-mediated CD4+T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis
Bin QING ; Xinxin KONG ; Dongfan YE ; Chuangye WANG ; Jian ZHANG ; Bin WANG ; Xiaoou HUANG ; Nanbo WANG ; Hang QIAN ; Zhi XU
Journal of Army Medical University 2025;47(19):2302-2314
Objective To investigate the characteristics of immune exhaustion in sepsis and analyze its association with gut microbiota translocation.Methods A total of 130 mice were randomly divided into a cecal ligation and puncture(CLP)group(n=100)and a Sham group(n=30)Mouse model of sepsis was established with CLP procedure.Flow cytometry was used to analyze the proportions of peripheral blood CD4+T and CD8+T cells and programmed cell death protein 1(PD-1)positive T cell subsets in mice.Bacterial colonization in organs such as the heart,liver and kidneys was quantified by plating homogenates of the organs.Pathological changes in immune organs were observed with HE staining.The expression and localization of CD4?,CD8?,and PD-1?cells in immune organs were detected with immunohistochemical staining,and Image J software was employed for subsequent quantification of the number of the positive cells.Results HE staining demonstrated that immune organs exhibited varying degrees of pathological damages with disease progression.Compared with the Sham mice,the CLP mice exhibited significantly increased bacterial colonization in parenchymal organs and peripheral blood(P<0.05),notably in the liver,which showed the most severe infection.In the CLP group,the proportion of CD4+T lymphocytes in peripheral blood at days 1,3,and 5 postoperatively was decreased by 56%,70.57%,and 87.42%,respectively,when compared with the Sham group(P<0.001).The proportion of CD8+T lymphocytes was decreased by 48.33%relative to the Sham group only at day 5(P<0.001).In contrast,the proportion of CD4+T cell subsets expressing PD-1 was increased to 673.08,423.08,and 600 times that of the Sham group,respectively,at the same postoperative time points(P<0.001).Immunohistochemical results showed that,in the CLP group,the proportion of CD4+T cells in the thymus,spleen,and mesenteric lymph nodes was increased to 7.65,2.66,and 3.7 times that of the Sham group,respectively,at the early-stage peak(P<0.001),and then these proportions were decreased by 82.8%(P<0.001),41.9%(P<0.01),and 60.15%(P<0.001),respectively,at the late-stage trough when compared with the early-stage peak in the corresponding organs.The proportion of CD8+positive cells was increased in the early stage and then decreased insignificantly,while the proportion of PD-1+positive cells was increased continuously,and reached 6.24,13.9,and 20.96 times that of the Sham group at the peak in the thymus,spleen,and mesenteric lymph nodes respectively(P<0.001),with their expression regions showing a rough overlap with those of CD4+cells.Conclusion During sepsis,the inflammatory response can cause severe damage to immune organs and persistent exhaustion of CD4?T lymphocytes,leading to declined defenses against infection,which may be the main causes for exacerbated gut microbiota translocation and then systemic infection.
4.Tubular gastric elongation surgery for high esophageal-gastric anastomosis after resection of esophageal cancer:analysis of 5 cases
Hai ZHONG ; Xiang LI ; Hongsen LIANG ; Nanbo LIU ; Yufan LIU ; Junhua ZHANG ; Xu WU
Journal of Southern Medical University 2018;38(1):123-125
Objective To summarize our experience with tubular gastric elongation surgery for management of insufficient gastric length for high esophageal-gastric anastomosis following esophageal carcinoma resection. Methods From September, 2015 to October 2016, 5 patients with esophageal cancer were treated in our department, including two with cervical esophageal cancer and 3 with thoracic esophageal cancer. The patients with cervical esophageal cancer underwent pharyngeal resection, total laryngectomy, esophageal varus extubation and gastric oropharyngeal anastomosis, and the patients with thoracic esophageal cancer underwent esophageal cancer resection with incisions on the left neck, the right chest and the median abdomen. During the surgery, the length of the stomach was found insufficient to allow routine oropharyngeal anastomosis, and tubular gastric elongation was conducted to extend the tubular stomach to enable successful completion of the surgery. Results All the patients recovered smoothly after the surgery and were discharged after 2-3 weeks. Conclusion Tubular gastric elongation surgery can be a good choice for high esophageal-gastric anastomosis after resection of esophageal cancer in cases of insufficient tubular stomach length or high tension at the anastomosis.
5.Tubular gastric elongation surgery for high esophageal-gastric anastomosis after resection of esophageal cancer:analysis of 5 cases
Hai ZHONG ; Xiang LI ; Hongsen LIANG ; Nanbo LIU ; Yufan LIU ; Junhua ZHANG ; Xu WU
Journal of Southern Medical University 2018;38(1):123-125
Objective To summarize our experience with tubular gastric elongation surgery for management of insufficient gastric length for high esophageal-gastric anastomosis following esophageal carcinoma resection. Methods From September, 2015 to October 2016, 5 patients with esophageal cancer were treated in our department, including two with cervical esophageal cancer and 3 with thoracic esophageal cancer. The patients with cervical esophageal cancer underwent pharyngeal resection, total laryngectomy, esophageal varus extubation and gastric oropharyngeal anastomosis, and the patients with thoracic esophageal cancer underwent esophageal cancer resection with incisions on the left neck, the right chest and the median abdomen. During the surgery, the length of the stomach was found insufficient to allow routine oropharyngeal anastomosis, and tubular gastric elongation was conducted to extend the tubular stomach to enable successful completion of the surgery. Results All the patients recovered smoothly after the surgery and were discharged after 2-3 weeks. Conclusion Tubular gastric elongation surgery can be a good choice for high esophageal-gastric anastomosis after resection of esophageal cancer in cases of insufficient tubular stomach length or high tension at the anastomosis.
6.Establishing the regeneration system of sunflower.
Yuanyuan WANG ; Caifeng LI ; Yifei ZHANG ; Yeting CHEN ; Liying ZHAO ; Peng YUE ; Xiangyong TENG ; Nanbo WANG
Chinese Journal of Biotechnology 2011;27(9):1379-1389
In order to establish a high efficient regeneration system of sunflower, we optimized the process of callus induction, differentiation and rooting by screening the optimum genotype, explant materials, hormone and cytokine concentration and additives. The results indicated that hybrid sunflowers were easier to regenerate than selfing ones; The best explant was four days cotyledon. The optimum induction medium was Murashige and Skoog (MS) + 2.0 mg/L 6-benzyladenine (6-BA) + 0.5 mg/L naphthaleneacetic acid (NAA) + 1.0 mg/L kinetin (KT). The maximum rate of callus induction was 100%. The optimum differentiation medium was MS +0.2 mg/L 6-BA + 0.5 mg/L NAA + 0.3 mg/L KT + 0.3 mg/L silver nitrate (AgNO3) + 0.2 g/L active carbon (AC), and the buds differentiation rate was up to 71%. The best rooting culture medium was 1/2 MS + 0.6 mg/L indolebutyric acid (IBA). The highest rooting rate was 77%. The analysis of variance showed that genotype, explants growth time, different kinds and concentration of hormone, AC concentration had a significant effect on sunflower regeneration.
Cotyledon
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drug effects
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growth & development
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Genotype
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Helianthus
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genetics
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physiology
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Hybridization, Genetic
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Plant Growth Regulators
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pharmacology
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Plant Leaves
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drug effects
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growth & development
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Plant Roots
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drug effects
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growth & development
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Regeneration
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drug effects
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physiology
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Seedlings
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drug effects
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growth & development
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Tissue Culture Techniques

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