BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

Objective To investigate the inhibitory effect of antimicrobial peptide WK-13-3D on the proliferation of triple negative breast cancer MDA-MB-231 cells and its potential mechanism.Methods The effect of antimicrobial peptide WK-13-3D at concentrations of 0,10,15,20,25,30,35,and 40 μmol/L on MDA-MB-231 cells proliferation was assessed using the CCK-8 assay.A pull-down assay was conducted to identify interacting proteins of antimicrobial peptide WK-13-3D with MDA-MB-231 cells.MDA-MB-231 cells were obtained and divided into the following groups:(1)control group and treatment groups with 10 and 20 μmol/L antimicrobial peptide WK-13-3D.Apoptosis was evaluated using flow cytometry and Western blotting was conducted to detect the expression change of heavy chain binding protein(BiP),protein kinase R-like endoplasmic reticulum kinase(PERK),eukaryotic translation initiation factor 2α(eIF2α),phosphorylated eIF2α(p-eIF2α),and Bax proteins within the cells.(2)Control group(transfected with no-load plasmid),si-BiP-592 group(transfected with si-BiP-592 interference plasmid)and si-BiP-592+WK-13-3D group(co-treated with si-BiP-592 interference plasmid and 10 μmol/L antimicrobial peptide WK-13-3D).Western blotting was used to detect the expression changes of BiP,PERK,eIF2α,p-eIF2α and Bax proteins.Twelve BALB/c mice were randomly divided into PBS group(n=4),taxol(TAX)group(n=4)and WK-13-3D group(n=4).All mice were subcutaneously injected with MDA-MB-231 cells to establish a triple-negative breast cancer transplant tumor model.WK-13-3D group received local injections of antimicrobial peptide WK-13-3D[200 mg/(kg·d)],TAX group was administered TAX intraperitoneally at the same dose[200 mg/(kg·d)],and PBS group was injected with an equivalent volume of PBS.Two weeks post-injection,the mice were killed,and the tumor weight and volume were measured and photographed.Immunohistochemistry staining was performed to evaluate the expressions of BiP and Ki-67 proteins in the tumor tissues.Results CCK-8 assay showed a gradual decrease in MDA-MB-231 cell survival rates with increasing concentrations of WK-13-3D,with an inhibitory concentration 50(IC50)of 19.82 μmol/L.The pull-down assay identified 268 interacting proteins of antimicrobial peptides and MDA-MB-231 cells,mainly including heavy-chain binding protein(BiP),heat shock protein 90 beta family member 1(HSP90B1),valerin-containing protein(VCP),heat shock cognate 71 kD protein(HSPA8).Compared with control group,treatment with 10 and 20 μmol/L antimicrobial peptide WK-13-3D significantly increased the apoptosis rate of MDA-MB-231 cells(P<0.05 or P<0.01),decreased BiP protein expression(P<0.05),and increased the expression levels of PERK,p-eIF2α,and Bax(P<0.05 or P<0.01),with no significant change in eIF2α protein expression(P>0.05).Compared with control group,si-BiP-592 group showed BiP protein expression significantly decreased(P<0.05),and the expression of PERK,p-eIF2α,and Bax proteins was significantly increased(P<0.05),with no significant change in eIF2α protein expression(P>0.05);Compared with si-BiP-592 group,si-BiP-592+WK-13-3D group showed a decrease in BiP protein expression(P<0.05)and an increase in PERK,p-eIF2α,and Bax protein expression(P<0.05 or P<0.01),with no significant change in eIF2α protein expression(P>0.05).Tumor volumes in mice treated with antimicrobial peptide WK-13-3D and TAX were significantly smaller than those in PBS group(P<0.05),and the immunohistochemical staining showed that the proportion of Ki-67 and BiP positive cells in tumor tissues of WK-13-3D treated mice was significantly lower than that in PBS group(P<0.01).Conclusion Antimicrobial peptide WK-13-3D could inhibit the proliferation of MDA-MB-231 cells and its mechanism may involve the activation of endoplasmic reticulum stress and the induction of cell apoptosis.

Objective To analyze the relationship between obesity indicators,muscle mass indices and hypertension in Dongxiang adults,and establish and evaluate a Nomogram model based on these indicators used to predict the risk of hypertension in this population.Methods A total of 1209 Dongxiang adults from Linxia Prefecture,Gansu Province were selected,11 obesity indicators and 5 muscle mass indicators,including neck circumference(NC),waist-to-hip ratio(WHR),BMI,ponderal index(PI),conicity index(CI),a body shape index(ABSI),body roundness index(BRI),abdominal volume index(AVI),hip index(HI),body adiposity index(BAI),and the ratio of limb fat mass to body weight(LFWR),appendicular skeletal muscle mass(ASM),appendicular skeletal muscle mass index(ASMI),appendicular skeletal muscle mass to BMI ratio(ASMBMI),skeletal muscle index(SMI),and trunk muscle mass to body weight ratio(TMWR)were measured.Logistic regression analysis was used to explore the relationship between each indicator and hypertension.Nomogram prediction model was constructed and validated by using R language.Results Among Dongxiang males,the NC,BMI,PI,WHR,CI,AVI,BRI,BAI,ASM,ASMI,and LFWR were lower in the normal and high-normal blood pressure groups compared to the hypertensive group,while SMI,ASMBMI,and TMWR were higher.Among females,similar trends were observed,with lower NC,BMI,PI,WHR,CI,AVI,BRI,BAI,and LFWR in the normal blood pressure and high-normal blood pressure groups compared to the hypertensive group,but SMI,ASMBMI,and TMWR higher than the hypertensive group(P＜0.05).Results of multivariate Logistic regression analysis indicated that age,NC,and WHR were risk factors for hypertension in Dongxiang adults,while was protective factor(P＜0.05).The area under curve(AUC)of the Nomogram model constructed based on these factors was 0.796,and the Bootstrap internal validation C-index was 0.7957,indicating good calibration of the model.Conclusion The Nomogram model constructed based on obesity and muscle mass indicators has good predictive efficiency for predicting the risk of hypertension in Dongxiang adults.

Objective To investigate influencing factors of textbook outcome(TO)in patients with pancreatic ductal adenocarcinoma undergoing minimally invasive pancreaticoduodenectomy(MIPD).Methods A retrospective analysis was conducted on the clinical data of 101 cases of pancreatic ductal adenocarcinoma treated with MIPD in our hospital from January 2020 to December 2022.According to the inclusion and exclusion criteria,89 cases were ultimately included in this study,of which 61 cases reached TO(TO group)and 28 cases did not reach TO(non-TO group).Variables with P<0.05 in univariate analysis were included in multivariate logistic regression analysis to identify independent prognostic factors of TO.Results Univariate analysis showed that there were significant differences in pancreatic duct dilation>3 mm,preoperative neutrophil lymphocyte ratio(NLR),extended hospital stay,postoperative hospitalstay,and drain fluid amylase(DFA)>1100 U/L at1-3 d aftersurgery(P<0.05).Multivariate logistic regression analysis showed that the independent prognostic factors affecting TO were:pancreatic duct dilation>3 mm(OR=7.290,95%CI:1.485-35.786,P=0.014),postoperative hospital stay(OR=0.862,95%CI:0.751-0.989,P=0.034),and the DFA on the first postoperative day>1100 U/L(OR=0.052,95%CI:0.005-0.545,P=0.014).Conclusions The outcome of TO in patients after MIPD is not related to the surgical approach(robot assisted minimally invasive pancreaticoduodenectomy or laparoscopic pancreaticoduodenectomy).Pancreatic duct dilation>3 mm,postoperative hospital stay,and DFA on the first postoperative day>1100 U/L are independent prognostic factors of TO after MIPD in patients with pancreatic ductal adenocarcinoma.

Objective:To evaluate the levels and source of cadmium exposure by dietary pathway among middle-aged and elderly people ≥40 in cadmium-contaminated areas of China.Methods:A total of 7 193 people aged 40-89 years from four typical cadmium-contaminated areas in China were selected as the study subjects. Food Frequency Questionnaire (FFQ), Total Diet Study (TDS) and a 3-day-24-hour dietary recall survey were conducted. Dietary cadmium intake and food sources through dietary pathways were assessed based on cadmium content in foods, consumption amounts and intake frequencies.Results:The mean age of the participants was 63.39±12.21 years, with 50.05% being males. The average monthly dietary cadmium intake was 7.39 μg/(kg·BW). Staple foods and vegetables were the primary sources of dietary cadmium intake, accounting for 57.51% and 32.48%, respectively. The monthly dietary cadmium intake in all surveyed regions did not exceed the Provisional Tolerable Monthly Intake (PTMI) recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA).Conclusion:The monthly dietary cadmium intake among middle-aged and elderly people in cadmium-contaminated areas of China is relatively low, with the risk remaining at an acceptable level. Staple foods and vegetables are the most significant contributors to dietary cadmium intake.

Aim To explore the effect of hydroxysafflor yellow A(HSYA)on lipocalin 2(LCN2)-induced fer-roptosis in HT22 cells and the related mechanism.Methods Thirty male Sprague-Dawley(SD)rats were used to establish the middle cerebral artery occlu-sion/reperfusion(MCAO/R)model by the suture method.The rats were randomly divided into the Sham group,the MCAO/R group,and the MCAO/R+HSYA group.The infarct area was measured by TTC staining,and the degree of neurological deficit was evaluated by the Z-Longa scoring method.The expressions of LCN2 and 24P3R in brain tissues were detected by Western blot.LCN2 protein was added to HT-22 cells,and the cells were divided into the normal group,the LCN2 group,and the LCN2+HSYA group.The optimal con-centration of LCN2-induced neuronal ferroptosis was screened by LDH assay and Western blot,and the ex-pression levels of ferritin,FPN1,GPX4,SLC7A11,COX2,and 24P3R were detected.LCN2 was knocked down by siRNA transfection,and the expressions of GPX4 and ferritin were detected.The contents of glu-tathione(GSH),malondialdehyde(MDA),GPX4,and Fe2+were determined by colorimetry,and the expres-sion of GPX4 was detected by immunofluorescence.The binding force between HSYA and LCN2 was ana-lyzed by molecular docking technology.Results Ani-mal experiments showed that HSYA could reduce the cerebral infarction area and decrease the neurological function score of MCAO/R rats.Compared with the sham group,the levels of LCN2 and 24P3R increased in the MCAO/R group,while HSYA inhibited their ex-pressions.Cell experiments showed that the optimal concentration of LCN2 to induce ferroptosis in HT22 cells was 2 μmol·L-1.After knocking down LCN2 by siRNA transfection,compared with the LCN2 group,the expression levels of GPX4 and ferritin in the siLCN2 group increased significantly.Compared with the nor-mal group,the expressions of SLC7A11,GPX4,FPN1,ferritin,and GSH in the LCN2 group decreased signifi-cantly,while the concentration of Fe2+,and the expres-sions of MDA,COX2,and 24P3R increased.HSYA could increase the expressions of SLC7A11,GPX4,FPN1,ferritin,and GSH,reduce the contents of Fe2+and MDA,and inhibit the expressions of COX2 and 24P3R.Molecular docking showed that the binding en-ergy between HSYA and LCN2 was-8.0 kJ·mol-1.Conclusion HSYA can inhibit LCN2-induced ferrop-tosis in HT22 cells through the SLC7A11/GPX4 signa-ling pathway.

In recent years, potato scab caused by pathogenic Streptomyces spp. has become widespread globally, with increasing damage severely compromising the commercial value and storability of tubers. The pathogens are transmitted through the soil and seeds of potato, while existing control technologies have demonstrated limited efficacy in preventing the colonization and spread of pathogens, which pose a critical bottleneck in the sustainable development of the potato industry. This study systematically examines the pathogen characteristics and pathogenic mechanisms, evaluates the impacts of soil nutrients and microbial community structure on disease severity, and analyzes limitations in current chemical control, biological control, and disease-resistant variety breeding approaches. We propose an integrated control strategy of disease-resistant varieties, phosphorus fertilizer reduction, fertilizer efficiency enhancement, and phosphorus-soluble antagonistic fungicides, aiming to provide novel research perspectives for achieving effective prevention and control of potato scab.
Solanum tuberosum/microbiology* ; Plant Diseases/prevention & control* ; Streptomyces/pathogenicity* ; Disease Resistance ; Fungicides, Industrial/pharmacology* ; Fertilizers ; Soil Microbiology

Poor prognosis and high mortality rate are frequently observed in patients with relapsed/refractory acute myeloid leukemia (R/R AML), and there is no standard salvage therapy for these patients. As a method to evade apoptosis, cancer cells often upregulate anti-apoptotic proteins BCL-2 and MCL-1. Recently, venetoclax-based combination therapies have demonstrated promising prospects in treating R/R AML. However, the prevalent use of venetoclax comes with a new challenge of resistance. Upregulation of BCL-1 and/or MCL-1 is the main cause of venetoclax resistance and preemptively targeting BCL-2/BCL-XL/MCL-1 can be used to delay or forestall drug resistance. Thus, selective targeting of BCL-2 and MCL-1 is a viable treatment strategy. This review reports the latest clinical progress on targeting BCL-2 and MCL-1 in R/R AML.
Humans ; Leukemia, Myeloid, Acute/drug therapy* ; Myeloid Cell Leukemia Sequence 1 Protein ; Proto-Oncogene Proteins c-bcl-2 ; Drug Resistance, Neoplasm ; Bridged Bicyclo Compounds, Heterocyclic/therapeutic use* ; Sulfonamides/therapeutic use* ; Recurrence