1.Effect of CD137-CD137 ligand pathway on phenotypic transformation of macrophages on wound healing of diabetic foot ulcer
Heng LI ; Cuilan ZHANG ; Yuze DUAN
Chinese Journal of Diabetes 2025;33(8):610-616
Objective To investigate whether CD137-CD137 ligand(CD137L)pathway affects foot ulcer wound healing in diabetic mice by regulating phenotypic transformation of macrophages.Methods A total of 24 db/db mice were randomly divided into model(Mod),CD137 and anti-CD137 groups with 8 mice in each group,and 8 db/m mice were selected as control(Con)group.Ulcer wounds were prepared in each group.The wound healing rate was calculated at the 7,14 and 21 day in each group.The pathological changes of the wound were evaluated by HE staining.The levels of serum IL-1β,TNF-α,VEGF and basic fibroblast growth factor(bFGF)were detected by ELISA,and the expression of wound angiogenesis marker CD31 was detected by immunofluorescence staining.The expression of the gene-induced nitric oxide synthase(iNOS),IL-6 in M1-type macrophages and arginase 1(Arg1),mannose receptor(CD206)in M2-type macrophages were detected by qRT-PCR.Flow cytometry was used to detect the proportion of M1 macrophages labeled F4/80 and CD86,and the proportion of M2 macrophages labeled F4/80 and CD206 in peripheral blood of mice in each group.Results Compared with Mod group,the skin tissue structure was more complete,inflammatory cell infiltration was reduced,new hair follicle tissues were increased,the wound healing rate of CD137 group at 7,14 and 21 d,VEGF,bFGF,the relative fluorescence intensity of CD31 in wound tissue,the mRNA expression of Arg1 and CD206,and the proportion of M2-type macrophages were increased(P<0.05),IL-1β,TNF-α,the mRNA expression of iNOS and IL-6 expression,and the proportion of M1-type macrophages in peripheral blood were decreased in CD137 group(P<0.05).Conclusions CD137-CD137L pathway may play a role in promoting wound healing in DFU mice by promoting the transformation of wound macrophage phenotype M1 to M2.
2.Effect of human milk on serum bilirubin and intestinal microbiota in neonatal jaundice rats
Qian PENG ; Heng ZHANG ; Shicheng JIANG ; Keyong LUO ; Miao DUAN
Chinese Journal of Comparative Medicine 2025;35(10):39-47
Objective To investigate the effects of human milk on serum bilirubin levels and gut microbiota in neonatal rats with hyperbilirubinemia.Methods A total of 24 7-day-old specific-pathogen-free Sprague-Dawley rats were injected with bilirubin or normal saline,respectively,and human milk or formula milk was administered 24 hours later for intervention.The rats were divided randomly into four groups:human milk-normal saline group(HN),human milk-bilirubin group(HB),formula milk-normal saline group(FN),and formula milk-bilirubin group(FB).Samples were taken 72 hours later,and serum bilirubin values were detected by enzyme-linked immunosorbent assay.The intestinal microbiota were analyzed using 16S rDNA high-throughput sequencing.Results There was no significant difference in bilirubin values among the groups.Pseudomonas was negatively correlated with indirect bilirubin value(P<0.05).The composition of the intestinal microbiota differed significantly between human milk and formula milk after gastric administration,with Firmicutes(P<0.01),Enterococci(P<0.05),being the main microbiota in the HN and HB groups,and Proteobacteria(P<0.001),Escherichia Shigella(P<0.01)and Acinetobacter(P<0.01)being the main in the FN and FB groups.Conclusions Pseudomonas may be negatively associated with bilirubin,and the structure of the intestinal microbiota may differ in relation to human milk and formula feeding.
3.Effect of CD137-CD137 ligand pathway on phenotypic transformation of macrophages on wound healing of diabetic foot ulcer
Heng LI ; Cuilan ZHANG ; Yuze DUAN
Chinese Journal of Diabetes 2025;33(8):610-616
Objective To investigate whether CD137-CD137 ligand(CD137L)pathway affects foot ulcer wound healing in diabetic mice by regulating phenotypic transformation of macrophages.Methods A total of 24 db/db mice were randomly divided into model(Mod),CD137 and anti-CD137 groups with 8 mice in each group,and 8 db/m mice were selected as control(Con)group.Ulcer wounds were prepared in each group.The wound healing rate was calculated at the 7,14 and 21 day in each group.The pathological changes of the wound were evaluated by HE staining.The levels of serum IL-1β,TNF-α,VEGF and basic fibroblast growth factor(bFGF)were detected by ELISA,and the expression of wound angiogenesis marker CD31 was detected by immunofluorescence staining.The expression of the gene-induced nitric oxide synthase(iNOS),IL-6 in M1-type macrophages and arginase 1(Arg1),mannose receptor(CD206)in M2-type macrophages were detected by qRT-PCR.Flow cytometry was used to detect the proportion of M1 macrophages labeled F4/80 and CD86,and the proportion of M2 macrophages labeled F4/80 and CD206 in peripheral blood of mice in each group.Results Compared with Mod group,the skin tissue structure was more complete,inflammatory cell infiltration was reduced,new hair follicle tissues were increased,the wound healing rate of CD137 group at 7,14 and 21 d,VEGF,bFGF,the relative fluorescence intensity of CD31 in wound tissue,the mRNA expression of Arg1 and CD206,and the proportion of M2-type macrophages were increased(P<0.05),IL-1β,TNF-α,the mRNA expression of iNOS and IL-6 expression,and the proportion of M1-type macrophages in peripheral blood were decreased in CD137 group(P<0.05).Conclusions CD137-CD137L pathway may play a role in promoting wound healing in DFU mice by promoting the transformation of wound macrophage phenotype M1 to M2.
4.Quality inspection of ultrasound soft tissue cutting hemostatic equipment
Jing HUANG ; Qi-di SUN ; Ao-wen DUAN ; Li XU ; Heng-yu LONG ; Hai-jiang ZHU ; He-hua ZHANG
Chinese Medical Equipment Journal 2025;46(10):49-53
Objective To carry out quality inspection of the ultrasound soft tissue cutting hemostatic equipment to ensure its safety and effectiveness.Methods Five brands of ultrasound soft tissue cutting hemostatic equipment were selected and noted as test equipment A,test equipment B,test equipment C,test equipment D and test equipment E,which underwent quality inspection in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current based on YY/T 0644-2008 Ultrasonics-surgical systems—Measurement and declaration of the basic output characteristics,YY/T 1750-2020 Ultrasonic surgical equipmetn for soft tissue excision and hemostasia and GB 9706.1-2020 Medical electrical equipment—Part 1:General requirements for basic safety and essential performance.Results The test data of the five brands in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current met the technical requirements of YY/T 0644-2008,YY/T 1750-2020,GB 9706.1-2020.Conclusion The quality inspection of the ultrasound soft tissue cutting hemostatic equipment contributes to enhancing the accuracy and stability of the equipment and decreasing the risk during its clinical application.[Chinese Medical Equipment Journal,2025,46(10):49-53]
5.Quality inspection of ultrasound soft tissue cutting hemostatic equipment
Jing HUANG ; Qi-di SUN ; Ao-wen DUAN ; Li XU ; Heng-yu LONG ; Hai-jiang ZHU ; He-hua ZHANG
Chinese Medical Equipment Journal 2025;46(10):49-53
Objective To carry out quality inspection of the ultrasound soft tissue cutting hemostatic equipment to ensure its safety and effectiveness.Methods Five brands of ultrasound soft tissue cutting hemostatic equipment were selected and noted as test equipment A,test equipment B,test equipment C,test equipment D and test equipment E,which underwent quality inspection in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current based on YY/T 0644-2008 Ultrasonics-surgical systems—Measurement and declaration of the basic output characteristics,YY/T 1750-2020 Ultrasonic surgical equipmetn for soft tissue excision and hemostasia and GB 9706.1-2020 Medical electrical equipment—Part 1:General requirements for basic safety and essential performance.Results The test data of the five brands in terms of tip main amplitude,tip lateral amplitude,tip vibration frequency,excitation frequency,static electrical power and contact current met the technical requirements of YY/T 0644-2008,YY/T 1750-2020,GB 9706.1-2020.Conclusion The quality inspection of the ultrasound soft tissue cutting hemostatic equipment contributes to enhancing the accuracy and stability of the equipment and decreasing the risk during its clinical application.[Chinese Medical Equipment Journal,2025,46(10):49-53]
6.IDENTIFICATION AND COMPOSITION OF MOSQUITO SPECIES IN EMEIFENG NATURE RESERVE,FUJIAN PROVINCE,CHINA
Jing-Wen XIE ; Ming WANG ; Yu-Tong DU ; Gang WANG ; Zi-Ge YIN ; Jing-Hong CAI ; Qing LIU ; Heng-Duan ZHANG ; Tong-Yan ZHAO ; En-Jiong HUANG
Acta Parasitologica et Medica Entomologica Sinica 2025;32(2):112-118
Objective In this preliminary survey,we sought to determine the composition of mosquito species inhabiting the Emeifeng Nature Reserve,Fujian Province,China.Methods Mosquito larvae were collected by straw and spoon trapping,and adult mosquitoes were collected by lamp trapping at selected breeding sites in the reserve.The specimens were initially identified based on morphology,with subsequent verification using molecular biology methods.Results A total of 34 mosquito species in 13 genera were collected,among which,there were 4 species of Anopheles(Genus Anopheles Meigen,1818),2 species of Lutzia(Genus Lutzia Theobald,1903),15 species of Culex(Genus Culex Linnaeus,1758),4 species of Stegomyia(Genus Stegomyia Theobald,1901),and single species of Hulecoeteomyia(Genus Hulecoeteomyia Theobald,1904),Luius(Genus Luius Reinert,Harbach et Kitching,2008),Aedes(Genus Aedes Meigen,1818),Downsiomyia(Genus Downsiomyia Vargas,1950),Collessius(Genus Collessius Reinert,Harbach et kitching,2006),Uranotaenia(Genus Uranotaenia Lynch 1891),Armigeres(Genus Armigeres Theobald,1901),Toxorhynchites(Genus Toxorhynchites Theobald,1901),and pestle mosquito(Genus Tripteroides Giles,1904).Conclusions The species composition of mosquitoes sampled in the Emeifeng Nature Reserve will provide a basis for further research on mosquito vectors and contribute to measures for local mosquito control.
7.Influence of serum alpha-fetoprotein and CA19-9 on the prognosis of patients with combined hepatocellular-cholangiocarcinoma after radical surgery
Hongjian ZHANG ; Xiaohui DUAN ; Heng ZHANG ; Jianpei XUYANG ; Yuhao QING ; Zicheng WANG ; Xianhai MAO
Chinese Journal of Hepatobiliary Surgery 2025;31(2):87-91
Objective:To analyze the impact of serum alpha-fetoprotein (AFP) and carbohydrate antigen 19-9 (CA19-9) on the prognosis of patients with combined hepatocellular-cholangiocarcinoma (CHC) after radical surgery.Methods:Clinical data of 100 patients diagnosed with CHC by postoperative pathology in Hunan Provincial People's Hospital from January 2009 to January 2019 were retrospectively analyzed, including 73 males and 27 females, aged (52.0±10.9) years. Univariate and multivariate Cox regression analysis were conducted to try to find out the effects of AFP and CA19-9 on postoperative disease-free survival (DFS) of patients with CHC. Subgroup analysis was also performed to analyze the DFS among patients with different levels of AFP and CA19-9. Kaplan-Meier method and log-rank test were used to plot and compare the survival curves between groups.Results:Univariate analysis showed that levels of AFP, CA19-9, alanine transaminase, aspartate transaminase, American Joint Committee on Cancer (AJCC) TNM staging, portal vein invasion, tumor number are associated with postoperative DFS in CHC patients (all P<0.05). The multivariate Cox analysis showed that AFP≥20.6 μg/L ( HR=6.686, 95% CI: 1.985-9.582), CA19-9≥35.2 U/L ( HR=5.790, 95% CI: 1.197-8.855), AJCC tumor TNM staging stage Ⅱ( HR=6.384, 95% CI: 2.069-11.532), and portal vein invasion ( HR=2.384, 95% CI: 1.100-10.125) were risk factors for a shortened DFS in CHC patients after surgery (all P<0.05). Subgroup analysis showed that patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L ( n=14) had a lower DFS compared to patients with AFP<20.6 μg/L and CA19-9≥35.2 U/L ( n=32), and patients with AFP≥20.6 μg/L and CA19-9<35.2 U/L ( n=20) ( χ2=6.23, 4.98, P=0.014, 0.043). Conclusion:AFP and CA19-9 are risk factors for DFS in CHC patients. Patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L showed a worse prognosis.
8.Effect of human milk on serum bilirubin and intestinal microbiota in neonatal jaundice rats
Qian PENG ; Heng ZHANG ; Shicheng JIANG ; Keyong LUO ; Miao DUAN
Chinese Journal of Comparative Medicine 2025;35(10):39-47
Objective To investigate the effects of human milk on serum bilirubin levels and gut microbiota in neonatal rats with hyperbilirubinemia.Methods A total of 24 7-day-old specific-pathogen-free Sprague-Dawley rats were injected with bilirubin or normal saline,respectively,and human milk or formula milk was administered 24 hours later for intervention.The rats were divided randomly into four groups:human milk-normal saline group(HN),human milk-bilirubin group(HB),formula milk-normal saline group(FN),and formula milk-bilirubin group(FB).Samples were taken 72 hours later,and serum bilirubin values were detected by enzyme-linked immunosorbent assay.The intestinal microbiota were analyzed using 16S rDNA high-throughput sequencing.Results There was no significant difference in bilirubin values among the groups.Pseudomonas was negatively correlated with indirect bilirubin value(P<0.05).The composition of the intestinal microbiota differed significantly between human milk and formula milk after gastric administration,with Firmicutes(P<0.01),Enterococci(P<0.05),being the main microbiota in the HN and HB groups,and Proteobacteria(P<0.001),Escherichia Shigella(P<0.01)and Acinetobacter(P<0.01)being the main in the FN and FB groups.Conclusions Pseudomonas may be negatively associated with bilirubin,and the structure of the intestinal microbiota may differ in relation to human milk and formula feeding.
9.Influence of serum alpha-fetoprotein and CA19-9 on the prognosis of patients with combined hepatocellular-cholangiocarcinoma after radical surgery
Hongjian ZHANG ; Xiaohui DUAN ; Heng ZHANG ; Jianpei XUYANG ; Yuhao QING ; Zicheng WANG ; Xianhai MAO
Chinese Journal of Hepatobiliary Surgery 2025;31(2):87-91
Objective:To analyze the impact of serum alpha-fetoprotein (AFP) and carbohydrate antigen 19-9 (CA19-9) on the prognosis of patients with combined hepatocellular-cholangiocarcinoma (CHC) after radical surgery.Methods:Clinical data of 100 patients diagnosed with CHC by postoperative pathology in Hunan Provincial People's Hospital from January 2009 to January 2019 were retrospectively analyzed, including 73 males and 27 females, aged (52.0±10.9) years. Univariate and multivariate Cox regression analysis were conducted to try to find out the effects of AFP and CA19-9 on postoperative disease-free survival (DFS) of patients with CHC. Subgroup analysis was also performed to analyze the DFS among patients with different levels of AFP and CA19-9. Kaplan-Meier method and log-rank test were used to plot and compare the survival curves between groups.Results:Univariate analysis showed that levels of AFP, CA19-9, alanine transaminase, aspartate transaminase, American Joint Committee on Cancer (AJCC) TNM staging, portal vein invasion, tumor number are associated with postoperative DFS in CHC patients (all P<0.05). The multivariate Cox analysis showed that AFP≥20.6 μg/L ( HR=6.686, 95% CI: 1.985-9.582), CA19-9≥35.2 U/L ( HR=5.790, 95% CI: 1.197-8.855), AJCC tumor TNM staging stage Ⅱ( HR=6.384, 95% CI: 2.069-11.532), and portal vein invasion ( HR=2.384, 95% CI: 1.100-10.125) were risk factors for a shortened DFS in CHC patients after surgery (all P<0.05). Subgroup analysis showed that patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L ( n=14) had a lower DFS compared to patients with AFP<20.6 μg/L and CA19-9≥35.2 U/L ( n=32), and patients with AFP≥20.6 μg/L and CA19-9<35.2 U/L ( n=20) ( χ2=6.23, 4.98, P=0.014, 0.043). Conclusion:AFP and CA19-9 are risk factors for DFS in CHC patients. Patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L showed a worse prognosis.
10.Mannitol inhibits the proliferation of neural stem cell by a p38 mitogen-activated protein kinase-dependent signaling pathway
Hai-Zhen DUAN ; Xin ZHOU ; Quan HU ; Meng-Long LIU ; Shu-Hong WANG ; Ji ZHANG ; Xu-Heng JIANG ; Tian-Xi ZHANG ; An-Yong YU
Chinese Journal of Traumatology 2024;27(1):42-52
Purpose::Mannitol is one of the first-line drugs for reducing cerebral edema through increasing the extracellular osmotic pressure. However, long-term administration of mannitol in the treatment of cerebral edema triggers damage to neurons and astrocytes. Given that neural stem cell (NSC) is a subpopulation of main regenerative cells in the central nervous system after injury, the effect of mannitol on NSC is still elusive. The present study aims to elucidate the role of mannitol in NSC proliferation.Methods::C57 mice were derived from the animal house of Zunyi Medical University. A total of 15 pregnant mice were employed for the purpose of isolating NSCs in this investigation. Initially, mouse primary NSCs were isolated from the embryonic cortex of mice and subsequently identified through immunofluorescence staining. In order to investigate the impact of mannitol on NSC proliferation, both cell counting kit-8 assays and neurospheres formation assays were conducted. The in vitro effects of mannitol were examined at various doses and time points. In order to elucidate the role of Aquaporin 4 (AQP4) in the suppressive effect of mannitol on NSC proliferation, various assays including reverse transcription polymerase chain reaction, western blotting, and immunocytochemistry were conducted on control and mannitol-treated groups. Additionally, the phosphorylated p38 (p-p38) was examined to explore the potential mechanism underlying the inhibitory effect of mannitol on NSC proliferation. Finally, to further confirm the involvement of the p38 mitogen-activated protein kinase-dependent (MAPK) signaling pathway in the observed inhibition of NSC proliferation by mannitol, SB203580 was employed. All data were analyzed using SPSS 20.0 software (SPSS, Inc., Chicago, IL). The statistical analysis among multiple comparisons was performed using one-way analysis of variance (ANOVA), followed by Turkey's post hoc test in case of the data following a normal distribution using a Shapiro-Wilk normality test. Comparisons between 2 groups were determined using Student's t-test, if the data exhibited a normal distribution using a Shapiro-Wilk normality test. Meanwhile, data were shown as median and interquartile range and analyzed using the Mann-Whitney U test, if the data failed the normality test. A p < 0.05 was considered as significant difference. Results::Primary NSC were isolated from the mice, and the characteristics were identified using immunostaining analysis. Thereafter, the results indicated that mannitol held the capability of inhibiting NSC proliferation in a dose-dependent and time-dependent manner using cell counting kit-8, neurospheres formation, and immunostaining of Nestin and Ki67 assays. During the process of mannitol suppressing NSC proliferation, the expression of AQP4 mRNA and protein was downregulated, while the gene expression of p-p38 was elevated by reverse transcription polymerase chain reaction, immunostaining, and western blotting assays. Subsequently, the administration of SB203580, one of the p38 MAPK signaling pathway inhibitors, partially abrogated this inhibitory effect resulting from mannitol, supporting the fact that the p38 MAPK signaling pathway participated in curbing NSC proliferation induced by mannitol.Conclusions::Mannitol inhibits NSC proliferation through downregulating AQP4, while upregulating the expression of p-p38 MAPK.

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