1.Bone marrow mesenchymal stem cell nanovesicles fusion neutrophil apoptotic bodies promote skin wound healing in diabetic mice
Zhanpeng SUN ; Sen LIU ; Ling SHI ; Kaiyuan CHEN ; Meichen SONG ; Yan WU ; Jing YU
Chinese Journal of Tissue Engineering Research 2026;30(1):34-42
BACKGROUND:Nanocell vesicles possess functions such as re-epithelialization,antioxidation,anti-inflammation,and regulation of extracellular matrix remodeling.Meanwhile,apoptotic bodies have the immunomodulatory effects.Therefore,the combination of the two to form nanofusion vesicles can synergistically promote the healing of diabetic skin wounds.OBJECTIVE:To elucidate the impact of nanofusion vesicles on skin wound healing in a diabetic murine model.METHODS:(1)Material preparation and characterization:The primary bone marrow mesenchymal stem cells of C57BL/6J neonatal mice and the neutrophil apoptotic bodies of C57BL/6J mice were isolated and extracted.The nanofusion vesicles were prepared by micro-extrusion mechanism.(2)In vitro experiment:MTT assay was used to detect the proliferative effect of different concentrations of nanofusion vesicles on NIH-3T3 cells and human umbilical vein endothelial cells.Reactive oxygen species fluorescence probe was used to detect the antioxidant effect of nano-fusion vesicles on NIH-3T3 cells treated with hydrogen peroxide(H2O2).The inhibitory effect of nanofusion vesicles on RAW 264.7 macrophage inflammation induced by lipopolyside was detected by real-time quantitative RT-qPCR.(3)In vivo experiment:36 male C57BL/6J mice were employed to develop a murine model of diabetes mellitus.Following the successful induction of diabetes,two circular full-thickness wounds,each with a diameter of 6 mm,were created on either side of the diabetic mice's spine using a skin punch.The mice were divided into three groups by random number table method.The control group was injected with 0.1 mL of phosphate buffer solution.The nanovesicle group was injected with 0.1 mL nanovesicles(25 μg/mL).The nanofusion vesicle group was injected with 0.1 mL nanofusion(25 μg/mL)vesicles.After treatment for three consecutive days,the wound healing and histomorphological changes were observed.RESULTS AND CONCLUSION:(1)In vitro experiment:nanofusion vesicles,when administered at concentrations ranging from 0 to 100 μg/mL,exhibited no toxic effects and promoted the proliferation of NIH-3T3 and HUVEC cell lines.Notably,a concentration of 25 μg/mL nanofusion vesicle significantly enhanced the proliferation of NIH-3T3 cells.Furthermore,the survival rate of human umbilical vein endothelial cells was observed to increase in correlation with escalating concentrations of nanofusion vesicles.Nanofusion vesicles had a good antioxidant effect.In comparison to the H2O2 group,the fluorescence signal indicative of reactive oxygen species was progressively diminished in both the nanovesicle group and the nanofusion vesicle group.Furthermore,nanofusion vesicles possessed anti-inflammatory capabilities,effectively mitigating the inflammatory response in macrophages triggered by lipopolysaccharide stimulation.(2)In vivo experiment:Hematoxylin-eosin and Masson's trichrome staining revealed that in comparison to the control group,both the nanovesicle group and the nanofusion vesicle group exhibited a significant increase in granulation tissue formation and collagen fiber deposition within the wounds by day 6.Notably,the nanofusion vesicle group displayed the most pronounced effects.On day 12,the wound of nanofusion vesicle group was significantly reduced,and the healing rate was significantly faster than that of other groups(P<0.01),and the effect of promoting wound healing was the most significant.Our findings demonstrated that nanofusion vesicles exhibited superior pro-cell proliferative,antioxidant,and anti-inflammatory properties,thereby exerting a beneficial effect on the promotion of skin wound healing in diabetic mouse models.
2.Bone marrow mesenchymal stem cell nanovesicles fusion neutrophil apoptotic bodies promote skin wound healing in diabetic mice
Zhanpeng SUN ; Sen LIU ; Ling SHI ; Kaiyuan CHEN ; Meichen SONG ; Yan WU ; Jing YU
Chinese Journal of Tissue Engineering Research 2026;30(1):34-42
BACKGROUND:Nanocell vesicles possess functions such as re-epithelialization,antioxidation,anti-inflammation,and regulation of extracellular matrix remodeling.Meanwhile,apoptotic bodies have the immunomodulatory effects.Therefore,the combination of the two to form nanofusion vesicles can synergistically promote the healing of diabetic skin wounds.OBJECTIVE:To elucidate the impact of nanofusion vesicles on skin wound healing in a diabetic murine model.METHODS:(1)Material preparation and characterization:The primary bone marrow mesenchymal stem cells of C57BL/6J neonatal mice and the neutrophil apoptotic bodies of C57BL/6J mice were isolated and extracted.The nanofusion vesicles were prepared by micro-extrusion mechanism.(2)In vitro experiment:MTT assay was used to detect the proliferative effect of different concentrations of nanofusion vesicles on NIH-3T3 cells and human umbilical vein endothelial cells.Reactive oxygen species fluorescence probe was used to detect the antioxidant effect of nano-fusion vesicles on NIH-3T3 cells treated with hydrogen peroxide(H2O2).The inhibitory effect of nanofusion vesicles on RAW 264.7 macrophage inflammation induced by lipopolyside was detected by real-time quantitative RT-qPCR.(3)In vivo experiment:36 male C57BL/6J mice were employed to develop a murine model of diabetes mellitus.Following the successful induction of diabetes,two circular full-thickness wounds,each with a diameter of 6 mm,were created on either side of the diabetic mice's spine using a skin punch.The mice were divided into three groups by random number table method.The control group was injected with 0.1 mL of phosphate buffer solution.The nanovesicle group was injected with 0.1 mL nanovesicles(25 μg/mL).The nanofusion vesicle group was injected with 0.1 mL nanofusion(25 μg/mL)vesicles.After treatment for three consecutive days,the wound healing and histomorphological changes were observed.RESULTS AND CONCLUSION:(1)In vitro experiment:nanofusion vesicles,when administered at concentrations ranging from 0 to 100 μg/mL,exhibited no toxic effects and promoted the proliferation of NIH-3T3 and HUVEC cell lines.Notably,a concentration of 25 μg/mL nanofusion vesicle significantly enhanced the proliferation of NIH-3T3 cells.Furthermore,the survival rate of human umbilical vein endothelial cells was observed to increase in correlation with escalating concentrations of nanofusion vesicles.Nanofusion vesicles had a good antioxidant effect.In comparison to the H2O2 group,the fluorescence signal indicative of reactive oxygen species was progressively diminished in both the nanovesicle group and the nanofusion vesicle group.Furthermore,nanofusion vesicles possessed anti-inflammatory capabilities,effectively mitigating the inflammatory response in macrophages triggered by lipopolysaccharide stimulation.(2)In vivo experiment:Hematoxylin-eosin and Masson's trichrome staining revealed that in comparison to the control group,both the nanovesicle group and the nanofusion vesicle group exhibited a significant increase in granulation tissue formation and collagen fiber deposition within the wounds by day 6.Notably,the nanofusion vesicle group displayed the most pronounced effects.On day 12,the wound of nanofusion vesicle group was significantly reduced,and the healing rate was significantly faster than that of other groups(P<0.01),and the effect of promoting wound healing was the most significant.Our findings demonstrated that nanofusion vesicles exhibited superior pro-cell proliferative,antioxidant,and anti-inflammatory properties,thereby exerting a beneficial effect on the promotion of skin wound healing in diabetic mouse models.
3.Status of anemia and iron deficiency among primary and secondary school students in Rural Nutrition Improvement Program areas of Guizhou Province in 2023
ZHU Shu, GUO Hua, LI Hongbo, SHI Zhu, WU Shengnan, HUANG Yiyanwen, SUN Yan, LIU Yiya
Chinese Journal of School Health 2026;47(2):178-182
:
To analyze the prevalence of anemia and iron deficiency among primary and secondary school students in Rural Nutrition Improvement Program areas of Guizhou Province in 2023, and to explore the related factors, so as to provide evidence for Rural Nutrition Improvement Program optimization.
Methods:
In September 2023, a stratified random cluster sampling strategy was used to select 40 rural compulsory education schools with rural nutrition improvement program in five counties of Guizhou Province. School level questionnaire was employed to collect information of basic characteristics and school meal implementation. A total of 7 826 primary and secondary school students aged 6-16 underwent anthropometry and hemoglobin (Hb) determination; serum ferritin (SF) was additionally measured in a random subsample of 1 795 pupils. Students in Grade 3 and above also completed a questionnaire covering demographic characteristics, dietary behaviours and nutrition knowledge. Group comparisons were conducted by Chi square test or Fisher s exact test, and multivariable Logistic regression models were constructed to identify factors associated with anemia and iron deficiency.
Results:
The overall Hb level was (133.21±12.95)g/L, with an anemia prevalence of 7.17%. The overall SF level was (69.58±59.01)μg/L, with an iron deficiency prevalence of 2.73%. Multivariable analysis showed that stunting ( OR =1.88), school menus without nutrient calculation ( OR =1.61) and absence of menu planning software in the current semester ( OR =2.34) independently increased anemia risk, whereas obesity reduced it ( OR =0.54) (all P <0.05). Girls ( OR =4.16) and Grades 7-9 ( OR =5.93) increased iron deficiency risk (both P <0.05). Compared with rarely eating fresh vegetables, students with consuming <3 kinds per day ( OR =0.08) or exactly 3 kinds per day ( OR =0.06) had lower iron deficiency risks (both P <0.05).
Conclusions
Anemia and iron deficiency are prevalent among primary and secondary school students in Guizhou. Targeted intervention measures should be implemented for key populations to enhance the effectiveness of nutrition improvement program.
4.Analysis of completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer : a national multicenter real-world study
Kexuan LI ; Tixian XIAO ; Xiaodong WANG ; Bin WU ; Guole LIN ; Yuchen GUO ; Ming QU ; Si WU ; Xiaodong YANG ; Yinshengbo′er BAO ; Baohua WANG ; Fan ZHANG ; Xiangwang YU ; Beizhan NIU ; Junyang LU ; Lai XU ; Guannan ZHANG ; Zhen SUN ; Guoyou ZHANG ; Yan SHI ; Hong JIANG ; Yongjing TIAN ; Yongxiang LI ; Hongwei YAO ; Jun XUE ; Quan WANG ; Lie YANG ; Qian LIU ; Yi XIAO
Chinese Journal of Digestive Surgery 2025;24(1):113-119
Objective:To investigate the completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients in the national multicenter real-world database.Methods:The prospective real-world study was conducted. The clinicopathological data of 1 074 patients who underwent surgical treatment for mid and low rectal cancer in 47 national medical institutions, including Peking Union Medical College Hospital et al, from May 12,2023 to May 11,2024 were collected. Observation indicators: (1) clinical characteristics of patients with mid and low rectal cancer; (2) initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer; (3) initial imaging evaluation of patients with mid and low rectal cancer; (4) imaging evaluation after neoadjuvant therapy for patients with mid and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3). Count data were described as absoluter numbers and/or percentages. Results:(1) Clinical characteristics of patients with mid and low rectal cancer. Of the 1 074 patients, there were 713 males and 361 females, aged 63(56,70)years. The body mass index of 1 074 patients was 24(21,26)kg/m 2.For American Society of Anesthesiologists classification, there were 147 cases of stage Ⅰ, 641 cases of stage Ⅱ, 157 cases of stage Ⅲ, 2 cases of stage Ⅳ, and there were 127 cases missing data. (2) Initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer. Of the 1 074 patients, there were 787 cases (73.28%) undergoing complete colonoscopy, and there were only 197 cases (18.34%) undergoing immunohistochemical evaluation of all four mismatch repair proteins. (3) Initial imaging evaluation of patients with mid and low rectal cancer. Of the 1 074 patients, there were 842(78.40%) patients completing magnetic resonance imaging (MRI) or ultrasound evaluation, and there were 914(85.10%) patients completing chest, abdomen, and pelvis enhanced computed tomography (CT) evaluation. In the 149 patients completing rectal ultrasound evaluation, there were 122 cases (81.88%) comple-ting T staging evaluation, and there were 81 cases (54.36%) completing N staging evaluation. In the 808 patients completing rectal MRI evaluation, there were 708 cases (87.62%) completing T staging evaluation, and there were 590 cases (73.02%) completing N staging evaluation. (4) Imaging evalua-tion after neoadjuvant therapy for patients with mid and low rectal cancer. Of the 388 patients with neoadjuvant therapy, there were 332 patients (85.57%) completing MRI or ultrasound evaluation, and there were 327 patients (84.28%) completing chest, abdomen, and pelvis enhanced CT evalua-tion. In the 70 patients completing rectal ultrasound evaluation, there were 65 cases (92.86%) com-pleting T staging evaluation, and there were 49 cases (70.00%) completing N staging evaluation. In the 327 patients completing rectal MRI evaluation, there were 246 cases (75.23%) completing T staging, and there were 228 cases (69.72%) completing N staging evaluation. Conclusion:The com-pletion rate of tumor imaging evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients on a national scale is relatively good.
5.Extraction process optimization,component analysis and biological activity evaluation for total polyphenols from Conioselinum vaginatu
Jun-long WANG ; Hui-jie YAN ; Yong-gang LIN ; Zi-wei LI ; Wen-pan SHI ; Sheng-qi JIANG ; Bin WU ; Qin-ze GU
Chinese Traditional Patent Medicine 2025;47(5):1449-1455
AIM To optimize the extraction process for total polyphenols from Conioselinum vaginatu(Spreng.)Thell.,make component analysis,and evaluate their anti-oxidant,hypoglycemic activities.METHODS The effects of ultrasound,enzymatic hydrolysis,acid hydrolysis,alcohol extraction and hydrolysis processes on the extraction quantity of total polyphenols were investigated,respectively.With extraction temperature,extraction time,ethanol concetration and liquid-solid ratio as influencing factors,extraction quantity of total polyphenols as an evaluation index,the extraction process was optimized by response surface method.HPLC was adopted in the identification of polyphenolic composition and determination of their contents.Subsequently,total polyphenols' scavenging capacities on DPPH,ABTS,OH free radicals,total reducing power and inhibitory capacity on α-glucosidase were determined.RESULTS The highest extraction quantity of total polyphenols was observable when extraction process was employed.The optimal conditions were determined to be 62 ℃ for extraction temperature,54 min for extraction time,69%for ethanol concentration,and 50∶1 for liquid-solid ratio,the extraction quantity of total polyphenols was(9.51±0.2)mg GAE/g.Seven constituents existed in C.vaginatu,among which ferulic acid demonstrated the highest content,followed by that of myricetin,while D-tryptophan content was the lowest.At the concentration of 7.61 mg/L,total polyphenols displayed the scavenging rates on DPPH,ABTS,OH free radicals of 80.70%,85.97%,28.60%,total reducing power of 0.22,and inhibition rate on α-glucosidase of 77.23%,respectively.CONCLUSION This stable and reliable method can be used for the extraction of total polyphenols from C.vaginatum with strong anti-oxidant,hypoglycemic activities.
6.Survival analysis of female breast cancer in Shanghai:a population-based study from 2002 to 2017
Chunxiao WU ; Yi PANG ; Kai GU ; Jiaying YAN ; Chunfang WANG ; Yongmei XIANG ; Yan SHI
China Oncology 2025;35(3):291-297
Background and purpose:The Shanghai Municipal Center for Disease Control and Prevention provides annual updates on cancer statistics in Shanghai.Breast cancer is one of the common malignant tumors among women.In recent years,the incidence of female breast cancer was increasing,while its trend of mortality showed declining.This study aimed to investigate the survival rates of new female breast cancer cases in Shanghai from 2002 to 2017.Methods:Data of new cases and deaths of female breast cancer patients with follow-up information from 2002 to 2017 were obtained from the Population-based Cancer Registry and Vital Statistics System of Shanghai Municipal Center for Disease Control and Prevention.Numbers,proportions,and survival rates were stratified by year of diagnosis,age,histological type and stage at diagnosis for analysis.The 5-year observed survival rates were calculated based on the life table method.The probabilities of surviving from 0 to 99 years were estimated with the Elandt-Johnson model,and then cumulative expected survival rates were calculated using the Ederer Ⅱ method.Finally,the 5-year relative survival rates were calculated.The annual percent change(APC)of survival rates was estimated by Joinpoint Regression Program.Results:A total of 73 600 new female breast cancer cases were diagnosed from 2002 to 2017 in Shanghai.Among them,67 681 cases were morphological verification,accounting for 91.96%.By December 31,2022,23 745(32.26%)cases had died,and 19 466(26.45%)cases had died of cancer.A total of 68 332(92.84%)cases,who were either dead or followed for over 5 years,were considered to have complete follow-up.The remaining 5 268(7.16%)cases were lost to follow-up.73 538(99.92%)cases were included in the observed cohort for survival analysis.The number of observed cases nearly doubled from 3330 in 2002 to 6095 in 2017.The 5-year observed survival rate changed from 78.77%in 2002 to 84.55%in 2017 dynamically,showed a low increasing trend with an average rate of 0.50%per year(APC=0.50%,t=8.75,P<0.001).The 5-year relative survival rate also increased from 83.46%to 89.24%slowly,with an average rate of 0.47%(APC=0.47%,t=9.80,P<0.001).The overall 5-year observation survival rate of female cancer was 83.24%(82.96%-83.52%),and the 5-year relative survival rate was 87.58%(87.29%-87.87%)in Shanghai from 2002 to 2017.It was increasing over time,decreasing with aging and advanced stage at diagnosis continuously.There was no significant difference in the 5-year relative survival rates between the groups aged 15 to 64(P>0.05).The group with an unknown stage had the highest number of cases,followed by the stage Ⅱ group,and then the stage Ⅰ group.The 5-year relative survival rate of cases with stage Ⅰ disease reached 99.10%(98.78%-99.42%),but these cases only accounted for 25.51%of the total.The 5-year relative survival rate of cases with stage Ⅳ disease was 52.54%(50.98%-54.11%),and these cases accounted for 6.13%of the total.The 5-year relative survival rate of cases with s unknown stage was 82.04%(81.42%-82.65%),and these cases accounted for 31.05%of the total.Conclusion:The diagnostic levels and survival rates of female breast cancer in Shanghai were relatively high and continue to improve.However,the proportions of cases with unknown histological type and unknown stage remain relatively high,and the proportion of stage Ⅰ cases is not very large.The survival rates of stage Ⅳ cases are relatively low.This study provides evidence for further research,prevention and control efforts for female breast cancer.
7.Relationship between Serum IGFBP-3,IGF-1 and Cardiac Function and Short-Term Prognosis in Patients with Dilated Cardiomyopathy Complicated with Heart Failure
Li-juan YUAN ; Shi-yuan XIE ; Yan ZHOU ; Fa-mao LI ; Yi-wu ZHENG
Progress in Modern Biomedicine 2025;25(9):1576-1583
Objective:To observe the relationship between serum insulin-like growth factor binding protein-3(IGFBP-3),insulin-like growth factor-1(IGF-1)and cardiac function and short-term prognosis in patients with dilated cardiomyopathy(DCM)complicated with heart failure(HF).Methods:102 patients with DCM complicated with HF were selected as study group and they were divided into good prognosis group and poor prognosis group based on prognosis,and 100 patients with simple DCM during the same period were selected as the control group.The cardiac function indicators[left ventricular end systolic volume(LVESV),left ventricular ejection fraction(LVEF),left ventricular end diastolic volume(LVEDV)]and serum IGFBP-3 and IGF-1 levels between the control group and the study group were compared.The correlation between cardiac function and IGFBP-3,IGF-1 was analyzed by pearson analysis.The serum IGFBP-3 and IGF-1 leveles between good prognosis group and poor prognosis group were compared,the correlation between prognosis and IGFBP-3,IGF-1 was analyzed by spearman.Receiver operating characteristic(ROC)curves were used to analyze the predictive value of serum levels of IGFBP-3 and IGF-1 alone and in combination for the short-term prognosis of DCM complicated with HF patients.Results:Compared with control group,study group had higher LVEDV,LVESV,IGFBP-3,IGF-1,and lower LVEF(P<0.05).Pearson analysis results showed that,IGFBP-3 and IGF-1 were positively correlated with LVEDV and LVESV,while which was negatively correlated with LVEF(P<0.05).Compared with good prognosis group,poor prognosis group had higher IGFBP-3,IGF-1(P<0.05).Spearman correlation analysis results showed that,the levels of IGFBP-3 and IGF-1 were positively correlated with the poor prognosis of DCM complicated with HF patients(P<0.05).The area under the curve(AUC)of serum IGFBP-3,IGF-1 levels,and their combined use for short-term prognosis of DCM complicated with HF patients were 0.715,0.749,and 0.831,respectively.Conclusion:Serum IGFBP-3 and IGF-1 levels are elevated in patients with DCM complicated with HF,and the changes in these two indicators are associated with poor cardiac function and prognosis in patients.The combined detection of IGFBP-3 and IGF-1 levels has a high predictive value for the short-term prognosis of DCM combined with HF.
8.Clinical Efficacy and Effects on Gastrointestinal Dynamics and Inflammatory Factors Levels of Combined Treatment with Shugan Qingwei Decoction Combined with Electromagnetic Stimulation for Reflux Esophagitis
Shi-wang CHEN ; Yan-long LI ; Zheng-quan WU
Progress in Modern Biomedicine 2025;25(9):1510-1516
Objective:To observe the clinical efficacy and Effects on gastrointestinal dynamics and inflammatory factors levels of combined treatment with electromagnetic stimulation and Shugan Qingwei Tang for reflux esophagitis(RE).Methods:142 cases of liver and stomach depression heat type RE patients who were treated in our hospital from April 2021 to May 2023 were selected.The patients were divided into control group(conventional drug therapy combined with electromagnetic stimulation)and study group(Shugan Qingwei decoction was added to the control group)by random number table method,with 71 cases each.The curative effect,gastrointestinal dynamics indexes,inflammatory factors and adverse reactions were observed between the two groups.Results:Compared with control group after treatment,the clinical total effective rate,serum gastrin(GAS),and motilin(MTL)were higher,while serum tumor necrosis factor-α(TNF-α),c-reactive protein(CRP),interleukin-1β(IL-1β),and interleukin-6(IL-6)were lower in the study group(P<0.05).There were no obvious adverse reactions between both groups.Conclusion:The combination therapy of Shugan Qingwei Tang and electromagnetic stimulation can improve the clinical treatment effect,improve gastrointestinal function,reduce the level of inflammatory factors,and has good safety for RE patients.
9.MXene nanoparticles Ti3C2Tx and photothermal effect promote wound healing in diabetic mice
Meiyun LI ; Sen LIU ; Kaiyuan CHEN ; Ling SHI ; Meichen SONG ; Jiahong CAO ; Yan WU ; Jing YU
Chinese Journal of Tissue Engineering Research 2025;29(28):6052-6060
BACKGROUND:MXene nanoparticles,due to their unique hydrophilicity,biocompatibility,and antibacterial properties,are widely used in wound,tumor,nerve repair,and cardiovascular treatments.However,it is still unclear what effect MXene nanoparticles have on diabetic wound healing.OBJECTIVE:To investigate the in vitro antioxidant,anti-inflammatory and photothermal antibacterial properties of MXene nanoparticles Ti3C2Tx as well as their effect on wound repair in diabetic mice.METHODS:(1)In vitro experiments:The cytotoxicity of Ti3C2Tx nanoparticles on mouse fibroblasts(NIH-3T3)at various concentrations was evaluated using the methyl thiazolyl tetrazolium(MTT)assay.NIH-3T3 cells were exposed to H2O2,and the MTT assay was used to detect the protective effects of different mass concentrations of Ti3C2Tx on NIH-3T3 cells.NIH-3T3 cells were exposed to H2O2,and the effect of Ti3C2Tx(20 μg/mL)on the generation of reactive oxygen species in NIH-3T3 cells was analyzed under illumination(or no illumination)treatment.RAW264.7 macrophages were divided into three groups:control group,lipopolysaccharide group,and lipopolysaccharide+Ti3C2Tx group.Real-time quantitative PCR was used to detect the expression of specific genes(CD86,interleukin 6,CD206,arginase 1)in the cells.Escherichia coli(or Staphylococcus aureus)were divided into three groups:control group,Ti3C2Tx group,and Ti3C2Tx illumination group.The bacterial survival rate was calculated by plate colony counting method.(2)In vivo experiments:Streptozotocin was administered intraperitoneally to ICR mice to induce a diabetic condition.After successful modeling,a full-thickness skin defect wound was created on the back of the mice using a circular punch.The experiment was divided into three groups:control group(n=6),Ti3C2Tx group(n=6),and Ti3C2Tx illumination group(n=6).The wound healing was observed,and CD31 and CD206 immunohistochemical staining of wound tissue was performed on day 7 after intervention.Hematoxylin-eosin staining and Masson staining of wound tissue were performed on days 7 and 14 after intervention.Ti3C2Tx solution was injected subcutaneously into ICR mice.After illumination(or non-illumination)exposure,the toxic effects of Ti3C2Tx on mice were analyzed by blood biochemical detection.RESULTS AND CONCLUSION:(1)In vitro experiments:Ti3C2Tx showed no cytotoxicity on NIH-3T3 cells at mass concentrations ranging from 5-160 μg/mL.It increased the survival rate of NIH-3T3 cells at a mass concentration of 20 μg/mL.Ti3C2Tx at 10-80 μg/mL significantly improved the survival rate of NIH-3T3 cells under H2O2 intervention.Ti3C2Tx significantly inhibited the generation of reactive oxygen species in NIH-3T3 cells under the intervention of H2O2,and illumination treatment further enhanced the effect of Ti3C2Tx on inhibiting the generation of reactive oxygen species.Ti3C2Tx effectively inhibited macrophage inflammation induced by lipopolysaccharide and promoted the transformation of cells into M2 macrophages with anti-inflammatory properties.Both Ti3C2Tx and Ti3C2Tx illumination significantly inhibited the growth of Escherichia coli and Staphylococcus aureus,and the inhibitory effect of Ti3C2Tx illumination was more significant.(2)In vivo experiments:Gross and histological analyses of the wound surface showed that both Ti3C2Tx and Ti3C2Tx illumination promoted wound healing in diabetic mice,and the promotion effect of Ti3C2Tx irradiation was more significant.Immunohistochemical staining results showed that both Ti3C2Tx and Ti3C2Tx illumination inhibited the inflammatory response in diabetic wounds and promoted angiogenesis,and the effect of Ti3C2Tx illumination was more significant.Blood biochemical test results showed that Ti3C2Tx and illumination had no obvious toxic effects on mice.(3)These results indicate that Ti3C2Tx nanoparticles efficiently promote the healing of skin wounds in a diabetic mouse model through antioxidation,anti-inflammation,and antibacterial actions via photothermal effects.
10.Angiopoietin-like protein 4 affects progression of diabetic foot by regulating fibroblast and endothelial cell functions
Qinghong SONG ; Nan WU ; Yan SHI ; Hongyu CUI ; Fei LIU ; Hanchong LIU ; Ning ZHOU ; Bin YAO
Chinese Journal of Tissue Engineering Research 2025;29(25):5396-5402
BACKGROUND:Studies have shown that vascular factors have an important impact on the occurrence of diabetic foot.OBJECTIVE:To investigate the important role of angiopoietin-like protein 4 in the formation of diabetic foot.METHODS:(1)Bioinformatics analysis was performed on the gene expression profile data of diabetic foot patients to find the key genes.The skin samples of diabetic foot patients and healthy people were collected for hematoxylin-eosin staining,immunohistochemical staining,and qRT-PCR experiments to detect the expression of angiopoietin-like protein 4.(2)The immortalized human skin fibroblast cell line and primary human umbilical vein endothelial cells were cultured.The two kinds of cells were divided into control group and exogenous angiopoietin-like protein 4 supplemented group.The migration ability and proliferation ability of fibroblasts were detected by scratch assay and CCK-8 assay.The proliferation ability of endothelial cells was detected by Ki67 assay.RESULTS AND CONCLUSION:(1)Bioinformatics analysis results showed that the down-regulation of angiopoietin-like protein 4 gene might be the key gene leading to the formation of diabetic foot.(2)Hematoxylin-eosin staining exhibited that compared with normal skin,angiopoietin-like protein 4 was weakly expressed in diabetic foot skin,and the mRNA level was decreased(P<0.01).(3)Scratch assay results demonstrated that compared with the control group,fibroblast migration ability was significantly enhanced in the angiopoietin-like protein 4 group.CCK-8 assay showed that the absorbance value of fibroblasts in the angiopoietin-like protein 4 group was higher than that of the control group at 24 and 48 hours(P<0.01,P<0.001).It is suggested that angiopoietin-like protein 4 can enhance the migration and proliferation of fibroblasts.(4)Ki67 assay results demonstrated that the number of Ki67 positive cells in the angiopoietin-like protein 4 group was significantly more than that in the control group.CCK-8 assay results showed that the absorbance value of endothelial cells in the angiopoietin-like protein 4 group was higher than that of the control group at 24 and 48 hours(P<0.05,P<0.001).(5)All findings suggest that angiopoietin-like protein 4 can enhance the proliferation of endothelial cells treated with high glucose.


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