Objective: The efficacy of antidepressants is influenced by a combination of genetic, individual, and environmental factors. This study aimed to investigate the association between the miR-182 rs76481776 polymorphism and the response to antidepressant treatment in major depressive disorder (MDD) patients, and its underlying molecular mechanisms. Methods: This study enrolled 180 MDD patients and 180 healthy controls. The rs76481776 genotype was determined using TaqMan-based qPCR. The severity of depression and treatment response were assessed using the Hamilton Depression Rating Scale (HAMD). The expression of miR-182 and BDNF was measured using RT-qPCR. The regulatory relationship between miR-182 and BDNF was confirmed through a luciferase reporter gene assay. The correlation between miR-182 and BDNF expression was evaluated using the Pearson correlation coefficient. Results: The T allele of rs76481776 was a significant risk factor for MDD (OR = 2.182, 95% CI: 1.424−3.345, p ＜ 0.001) and was significantly associated with higher baseline scores on the HAMD. Moreover, individuals carrying the T allele (CT/TT genotype) exhibited a significantly poorer response to antidepressant treatment and a lower remission rate within 12 months compared to those with the CC genotype (p ＜ 0.05). Mechanistically, miR-182 was highly expressed in patients with MDD (p ＜ 0.05), and its expression was even higher in T allele carriers (p ＜ 0.05). miR-182 could directly target and suppress BDNF, leading to decreased BDNF expression in MDD patients, and its expression was significantly and inversely correlated with BDNF expression. Conclusion The T allele of rs76481776 diminished the therapeutic efficacy of antidepressants by up-regulating miR-182 expression and subsequently suppressing BDNF expression.

Objective:To investigate the differences in electrophysiological brain development of neonates in Tibet and Beijing.Methods:This prospective cohort study included neonates with gestational ages of 28 to 40 weeks and 6 days, without asphyxia, hypoxia, or brain injury, who were born between January 2022 and June 2024 at the Tibet Autonomous Region People's Hospital and Peking University First Hospital. The first electroencephalographic (EEG) monitoring was completed within 48 hours to 7 days after birth, which included a 4-channel amplitude-integrated EEG (aEEG) and a 12-channel continuous EEG (cEEG). Two electrophysiology experts scored the EEG results according to a rating scale, and the intraclass correlation coefficient (ICC) was used to explore the consistency between different evaluators. Preterm infants with gestational ages of 32 to 36 weeks and 6 days and post-menstrual age (PMA) less than full-term at the first EEG monitoring were re-examined with aEEG and cEEG at PMA of 37 to 40 weeks and 6 days. Infants were grouped based on PMA at the first EEG monitoring. Spearman rank correlation was used to analyze the correlations between total aEEG+cEEG scores, individual aEEG and cEEG scores, and PMA, gestational age, birth weight, and head circumference at the first EEG monitoring. Mann-Whitney U test, Kruskal-Wallis H test, and Bonferroni correction were used to compare the differences in total aEEG+cEEG scores, individual aEEG and cEEG scores between Tibet and Beijing, among adjacent PMA groups, and for premature infants at full-term PMA. Results:(1) A total of 341 neonates were included in this study, including 154 cases from Tibet (nine cases in the PMA of 28-29 weeks and 6 days group, 13 cases in the PMA of 30-31 weeks and 6 days group, 28 cases in the PMA of 32-33 weeks and 6 days group, 38 cases in the PMA of 34-36 weeks and 6 days group, and 66 cases in the PMA of 37-40 weeks and 6 days group) and 187 cases from Beijing (10 cases in the PMA of 28-29 weeks and 6 days group, 10 cases in the PMA of 30-31 weeks and 6 days group, 16 cases in the PMA of 32-33 weeks and 6 days group, 91 cases in the PMA of 34-36 weeks and 6 days group, and 60 cases in the PMA of 37-40 weeks and 6 days group). (2) Inter-rater consistency:the consistency of PMA inferred based on the total aEEC+CEEC score and actual PMA was high in two raters ( ICCrater one=0.96, ICCrater two=0.94, both P<0.01). (3) The correlation between total aEEG+cEEG score and PMA ( r=0.80) was stronger than that between the aEEG alone or cEEG scores and PMA ( r were 0.79 and 0.66, respectively). The total aEEG+cEEG score also correlated with gestational age at birth ( r=0.74), birth weight ( r=0.69), and head circumference at first EEG monitoring ( r=0.69) (all P<0.01). (4) Regardless of whether in Tibet or Beijing, the total aEEG+cEEG score increased sequentially in the PMA of 30- 31 weeks and 6 days, 32-33 weeks and 6 days, 34-36 weeks and 6 days, and 37-40 weeks and 6 days groups; the cEEG score increased sequentially in the PMA of 32-33 weeks and 6 days group, 34-36 weeks and 6 days group, and 37-40 weeks and 6 days groups; the aEEG score in the PMA 32- 33 weeks and 6 days group was higher than that in the 30-31 weeks and 6 days group, and the score in the PMA 37-40 weeks and 6 days group was higher than that in the 34-36 weeks and 6 days group (Bonferroni correction, all P<0.05). (5) At PMA of 34-36 weeks and 6 days, the total aEEG+cEEG score [25 points (22-26 points) vs. 26 points (24-28 points), Z=－2.62, P=0.009] and cEEG score [12 points (12-14 points) vs. 15 points (13-16 points), Z=－4.77, P<0.001] of newborns in Tibet were lower than those in Beijing, while the aEEG score was higher than those in Beijing [12 points (10-13 points) vs. 11 points (10-12 points), Z=2.17, P=0.030]; at PMA of 37-40 weeks and 6 days, the cEEG score of newborns in Tibet was lower than those in Beijing [16 points (15-17 points) vs. 17 points (15-18 points), Z=－2.27, P=0.023]. (6) The total aEEG+cEEG score of preterm infants born at 32 to 33 weeks and 6 days in Tibet was lower at PMA full-term compared to those in Beijing [27 points (26-28 points) vs. 29 points (28 -30 points), Z=－2.94], and also lower compared to the total aEEG+cEEG score of full-term gestational age newborns in Tibet during their first EEG monitoring [29 points (27-30 points)] (both P<0.05). Conclusions:In the high-altitude hypobaric hypoxic environment, the electroencephalographic development of newborns, especially premature infants, maybe lag behind of plain areas. The combined use of aEEG+cEEG may provide a better evaluation of neonatal brain development than using cEEG or aEEG alone.

Objective:To compare the efficacy of body posture combined with pharmacotherapy and pharmacotherapy alone in the treatment of chronic subdural hematoma(CSDH).Methods:Firstly, retrospective case series study was conducted. Thirty cases of CSDH that had received body posture combined with pharmacotherapy at Department of Neurosurgery, Huashan Hospital Affiliated to Fudan University from December 2016 to October 2020 were studied retrospectively. Twenty-seven patients were male, and 3 patients were female. The age of patients ( M(IQR)) was 66(16) years (range:28 to 84). Nineteen patients had unilateral hematoma, and 11 patients had bilateral hematoma. All patients received pharmacotherapy and body posture therapy that was to raise their lower limbs 20 to 30 cm with leg lift pad and get abdominal compressed with customized abdominal belt in supine position. Patients were required to maintain the body posture as much as possible, with the maximum to 16 to 18 hours per day. Patients with unilateral hematoma should tilt the head to the affected side and avoid tilting it to the opposite side. For patients with bilateral hematoma, there was no need for head lateralization. Patient were treated with oral dexamethasone and atorvastatin simultaneously. The preliminary efficacy of body posture combined with pharmacotherapy was determined by hematoma improvement rate which was analyzed by Clopper-Pearson method. Then, the multi-center, prospective, randomized controlled trial had carried out in 9 medical centers from August 2020 to November 2021. The stratified block randomization method was adopted. Patients were randomized in a ratio of 1∶1 to either receive pharmacotherapy alone(the control group) or body posture combined with pharmacotherapy(the experiment group) for 3 months and followed up for 6 months. Effective treatment was defined as complete absorption of hematoma, or the hematoma volume decreased by more than 10 ml and Markwalder grading scale score had improved by more than 1 point compared to the baseline. The efficacy rate and surgery conversion rate at 3 months and recurrence at 6 months were observed. Comparison between groups was performed with paired sample t test, Mann-Whitney U test, χ2 test, corrected χ2 test, or Fisher exact probability method. Logistic regression was used to compare the effective rate and operation rate between the two groups. Results:In the respective study, 30 patients completed follow-up 13 to 353 days after treatment. At the last follow-up, the incidence of almost complete absorption or significantly absorption of hematoma (hematoma volume was significantly reduced accompanied by symptom improvement) was 93.3%. The 95% CI for the incidence that analyzed by the Clopper-Pearson method was 77.9% to 99.2%. One hundred and six patients were enrolled in the multicenter study. Fifty-five patients underwent body posture combined with pharmacotherapy. The age was 74(17) years (range:26 to 92). Thirty-nine patients were males and 16 were females. Fifty-one patients underwent pharmacotherapy alone. The age was 69(12) years (range:48 to 84). Thirty-seven patients were males and 14 were females. The length of body posture recorded in diary card was (15.7±2.3) hours(range:7.6 to 19.3 hours). The efficacy rate in the body posture combined with pharmacotherapy group and pharmacotherapy alone group were 83.6% (46/55) and 56.9% (29/51), respectively at 3 months. The result of the logistic regression analysis showed that the efficacy of body posture combined with pharmacotherapy group was better than that of pharmacotherapy alone group ( OR=3.88,95% CI:1.57 to 9.58, P=0.003). Surgery rate in the body posture combined with pharmacotherapy group and pharmacotherapy alone group were 5.5% (3/55) and 21.6% (11/51) respectively. The result of Logistic regression showed that the pharmacotherapy alone group was more likely to be converted to surgery ( OR=0.21,95% CI:0.05 to 0.80, P=0.023). At the 6 months, no recurrence of cases was found in the body posture combined with pharmacotherapy group. However, the recurrence rate of pharmacotherapy alone group was 6.3% (3/48), there was no significant difference between the two groups ( P>0.05). Conclusion:The effect of body posture combined with pharmacotherapy for chronic subdural hematoma is better than that of pharmacotherapy alone.

Objective:To compare the preliminary efficacy and adverse events of chemoradiotherapy (CRT) with or without immunotherapy in bladder preservation therapy for localized muscle-invasive bladder cancer (MIBC) confined to the pelvis.Methods:Clinical data of 60 patients with MIBC who received CRT with or without immunotherapy for bladder preservation at the Cancer Hospital, Chinese Academy of Medical Sciences from January 2016 to June 2024 were retrospectively analyzed. Patients were divided into CRT plus immunotherapy group and CRT-alone group. Survival outcomes, bladder function preservation, recurrence and metastasis, as well as early and late radiation toxicities were evaluated. The Mann-Whitney U test was used for between-group comparisons. Overall survival (OS), progression-free survival (PFS), local recurrence-free survival (LRFS), and distant metastasis-free survival (DMFS) were estimated by the Kaplan-Meier method, and survival rates were compared by the log-rank test. Results:In the CRT plus immunotherapy group ( n=23), the median follow-up was 20 months. The median OS and median PFS were not reached. The 2-year OS, PFS, LRFS, and DMFS rates were 95.7%, 70.7%, 70.7%, and 92.9%, respectively, and 22 patients (96%) preserved normal bladder function. Patients with programmed death-ligand 1 (PD-L1) combined positive score (CPS) ≥1 had significantly higher 1-year PFS rate than those with CPS <1 (100% vs. 66.7%, P=0.004). In the CRT-alone group ( n=37), the median follow-up was 37 months, with median OS and PFS of 68 and 19 months, respectively. The 2-year OS, PFS, LRFS, and DMFS rates were 92.0%, 41.1%, 60.9% and 81.5%, respectively, and 33 patients (89%) preserved normal bladder function. Compared with the CRT-alone group, the CRT plus immunotherapy group showed a significant improvement in PFS ( χ2=4.38, P=0.036), while no significant differences were observed in OS, LRFS, or DMFS (all P>0.05). The incidence of acute hematologic toxicity in the CRT plus immunotherapy group and CRT-alone group were 52% (12/23), 27% (10/37) respectively, and late genitourinary toxicity was 22% (5/23), 8% (3/37), respectively, with no significant differences in overall acute or late toxicities (all P>0.05). Conclusions:For localized MIBC, bladder preservation with CRT combined with immunotherapy significantly improves PFS compared with CRT alone, while maintaining comparable safety. The PD-L1 status may serve as a favorable predictor for immunotherapy efficacy.

Objective:To establish 30-day of age transcutaneous bilirubin (TcB) reference curves for healthy neonates, and to investigate regional variations in bilirubin dynamics.Methods:A multicenter retrospective cohort study was conducted. A total of 220 950 healthy neonates born at a gestational age of 35-<42 weeks, with a birth weight ≥2 000 g, who did not receive phototherapy within 60 h after birth were recruited. All of them underwent remote TcB monitoring using the Bilibaby remote jaundice monitoring system between August 1 st, 2020 and December 31 st, 2024 in 426 hospitals. TcB data were collected within the period from birth to 30-day of age. The P40, P75, and P95 of TcB values were calculated, and dynamic TcB curves for 30-day of age were constructed. Patterns of bilirubin change, rates of change, and transition outcomes were described. Regional comparisons between South and North were conducted using linear mixed-effects models for TcB trajectories and Pearson′s chi-square test for outcome differences. Results:A total of 220 950 neonates were included, of whom 101 711 (46.03%) were female. Gestational age at birth was (38.75±1.12) weeks, and birth weight was (3 272±417) g. TcB levels increased rapidly within 3-day of age, peaked at 4-6-day of age, with peak values at P40, P75, and P95 of 200.6, 239.7 and 275.4 μmol/L (11.8, 14.1 and 16.2 mg/dl), respectively. TcB levels gradually declined thereafter and stabilized after 13-day of age, with values at P40, P75, and P95 fluctuating between 147.9-159.8, 190.4-200.6, and 231.2-239.7 μmol/L (8.7-9.4, 11.2-11.8, 13.6-14.1 mg/dl), respectively. Notably, among neonates categorized as low-or low-intermediate-risk within 3-day of age, 6 700 (12.76%) progressed to intermediate-high or high risk between 4 and 30 days of age. Before 13-day of age, TcB levels in the southern regions were consistently higher than those in the northern regions ( P=0.039); from 14 to 30 days of age, the overall TcB levels had no statistically difference, but the temporal changes in TcB still showed regional differences (degrees of freedom=3, all interaction P<0.05). Among neonates classified as low-or low-intermediate risk within 3-day of age, 25 326 were from southern regions, of whom 4 254 (16.80%) progressed to intermediate-high or high risk between 4 and 30 days of age. In northern regions, 27 193 neonates were classified as low-or low-intermediate risk within 3-day of age, among whom 2 446 (8.99%) progressed to intermediate-high or high risk. The risk progression between the 2 regions had statistically difference ( χ2=716.49, P<0.001). Conclusions:A TcB percentile curve for neonates within 30-day of age was established, revealing that both the overall TcB level and its temporal trend were higher in southern than in northern newborns. These findings provide baseline data to support continuous management of neonatal jaundice.

Objective To explore the clinical value of inflammatory prognostic index combined with dual-source CT perfusion imaging in predicting early response of hepatocellular carcinoma(HCC)after transcatheter hepatic artery chemoembolization(TACE).Methods A total of 25 patients with HCC,who met the inclusion criteria and received initial TACE at the First Affiliated Hospital of Dali University of China from November 2022 to November 2023,were prospectively collected.CT perfusion scan was performed before TACE as well as in 30-40 days after TACE,and blood routine and blood biochemical data were collected.The modified Response Evaluation Criteria in Solid Tumors(mRECIST)was used to evaluate postoperative enhanced CT manifestations.Patients obtaining complete remission(CR)or partial remission(PR)were classified as effective group(n=14),and patients obtaining stable disease(SD)or progression disease(PD)were classified as ineffective group(n=11).The differences in CT perfusion imaging parameters and inflammatory parameters between the two groups were analyzed,and the predictive values of preoperative CT perfusion imaging parameters and inflammatory parameters for postoperative early response of HCC were evaluated.The cutoff value was taken at the maximum Youden index.Univariate analysis and multivariate analysis were used to analyze the effect of CT perfusion imaging parameters,inflammatory parameters and clinical features on the prognosis.The nomogram prediction model was constructed by using R software.Results The post-TACE arterial liver perfusion(ALP),hepatic perfusion index(HPI),blood flow(BF)and blood volume(BV)were significantly lower than their pre-TACE values(all P＜0.05).Afer TACE portal vein perfusion(PVP)was obviously higher than that before operation(P＜0.05).No statistically significant differences in the mean transit time(MTT),flow extraction product(FED),C-reactive protein(CRP),albumin(ALB),neutrophil/lymphocyte ratio(NLR)and inflammatory prognosis index(IPI)existed between the pre-TACE values and post-TACE values(all P＞0.05).The pre-TACE ALP,BF and FED in the effective group were significantly higher than those in the ineffective group,while the pre-TACE CRP and IPI in the effective group were remarkably lower than those in the ineffective group(P＜0.05).There were no statistically significant differences in pre-TACE PVP,HPI,MTT,BV,ALB and NLR between the effective group and the ineffective group before(all P＞0.05).In the effective group,the pre-TACE ALP,HPI,BF,BV and FED were obviously higher than their post-TACE values(all P＜0.05)preoperative PVP was significantly lower than postoperation(P＜0.05),while no statistically significant differences in MTT,CRP,ALB,NLR and IPI existed between the pre-TACE values and the post-TACE values(all P＞0.05).In the ineffective group,the pre-TACE HPI was prominently higher than the post-TACE value(P＜0.05),while no statistically significant differences in ALP,PVP,BF,BV,MTT,FED,ALB,CRP,NLR and IPI existed between the pre-TACE values and the post-TACE values(all P＞0.05).The pre-TACE ALP,BF,BV,FED,CRP and IPI had high predictive values in judging early response of HCC after TACE(all P＜0.05),the AUC values were 0.831,0.779,0.740,0.753,0.779 and 0.805 respectively,and the optimal cutoffs were 33.280 mL/100 mL min,61.860 mL/100 mL min,5.885 mL/100 mL,29.725 mL/100 mL min,30.465 mg/L,1.885 respectively.ALP combined with CRP had the highest predictive value for post-TACE early response of HCC,with an AUC of 0.968(95％CI:0.906-1.000,P＜0.05),ALP combined with IPI could significantly improve the predictive value,with an AUC of 0.961(95％CI:0.894-1.000,P＜0.05),with the sensitivity and specificity being 0.929 and 0.909 respectively.Multivariate analysis showed that pre-TACE ALP and CRP were the independent influencing factors for post-TACE early response of HCC(P＜0.05).The nomogram prediction model constructed based on the pre-TACE ALP and CRP could effectively predict the post-TACE early response of HCC,and the AUC value was 0.968(95％CI:0.908-1.000).Conclusion ALP and CRP can be used to predict the post-TACE early response of HCC,and the combination use of ALP and CRP can significantly improve the predictive value.

Objective To investigate the therapeutic effect of gabapentin combined with hemoperfusion on skin pruritus in the patients on maintenance hemodialysis(MHD).Methods A total of 72 patients with MHD complicating moderate to severe skin itching treated in Lianyungang Municipal Oriental Hospital from January 2023 to March 2024 were selected as the study subjects and divided into the observation group and control group by the random number table method,36 cases in each group.On the basis of symptomatic treat-ment,the control group was given hemodialysis 3 times a week and hemoperfusion twice a month;on the basis of the control group,the observation group orally took 1 gabapentin capsule every night,and the patients in the two groups continued to be treated for 12 weeks.The visual analogue scale(VAS)score,Pittsburgh Sleep Quality Index(PSQI)score,molecular toxin level and calcium and phosphorus metabolism were evaluated be-fore treatment,in 4,12 weeks after treatment respectively,and the occurrence of adverse reactions during treatment in the two groups was observed.Results After 4 weeks and 12 weeks of treatment,the VAS and PSQI scores of the observation group were decreased compared with those before treatment,moreover the scores of the observation group were lower than those of the control group in the same period,and the differ-ences were statistically significant(P<0.05);the VAS and PSQI scores after 4 weeks of treatment in the control group were decreased compared with those before treatment,but the difference was not statistically significant(P>0.05);and the VAS and PSQI scores after 12 weeks of treatment in the control group were decreased compared with before treatment,and the difference was statistically significant(P<0.05).After 4 weeks of treatment,the levels of Scr,BUN and β2-MG in the two groups were decreased compared with those before treatment,but the differences were not statistically significant(P>0.05);after 12 weeks of treatment,the levels of Scr,BUN and β2-MG in the two groups were decreased compared with those before treatment and after 4 weeks of treatment,and the differences were statistically significant(P<0.05),but there was no sta-tistically significant difference between the two groups(P>0.05).After 4 weeks of treatment,there was no statistically significant difference in the levels of blood Ca,P and iPTH between the two groups compared with before treatment(P>0.05);after 12 weeks of treatment,except for blood Ca,the levels of blood P and iPTH in the two groups were decreased compared with those before treatment and in 4 weeks of treatment(P<0.05),and the differences were statistically significant(P<0.05),but there was no statistical difference be-tween the two groups(P>0.05).There were no adverse reactions in the control group,and there were 3 cases of drowsiness,1 case of fatigue and 1 case of dizziness at the beginning of treatment in the observation group.The symptoms were mild without treatment and spontaneously relieved in about 1 week.Conclusion Gabap-entin combined with hemoperfusion could quickly and effectively alleviate the itchy symptoms of skin,improve the sleep quality,reduce the uremic toxin levels,and improve the calcium and phosphorus metabolism in MHD patients,with few adverse reactions,and the patients are easy to tolerate.

BACKGROUND: Chronic kidney disease (CKD) is associated with common pathophysiological processes, such as inflammation and fibrosis, in both the heart and the kidney. However, the underlying molecular mechanisms that drive these processes are not yet fully understood. Therefore, this study focused on the molecular mechanism of heart and kidney injury in CKD. METHODS: We generated an microRNA (miR)-26a knockout (KO) mouse model to investigate the role of miR-26a in angiotensin (Ang)-II-induced cardiac and renal injury. We performed Ang-II modeling in wild type (WT) mice and miR-26a KO mice, with six mice in each group. In addition, Ang-II-treated AC16 cells and HK2 cells were used as in vitro models of cardiac and renal injury in the context of CKD. Histological staining, immunohistochemistry, quantitative real-time polymerase chain reaction (PCR), and Western blotting were applied to study the regulation of miR-26a on Ang-II-induced cardiac and renal injury. Immunofluorescence reporter assays were used to detect downstream genes of miR-26a, and immunoprecipitation was employed to identify the interacting protein of LIM and senescent cell antigen-like domain 1 (LIMS1). We also used an adeno-associated virus (AAV) to supplement LIMS1 and explored the specific regulatory mechanism of miR-26a on Ang-II-induced cardiac and renal injury. Dunnett's multiple comparison and t -test were used to analyze the data. RESULTS: Compared with the control mice, miR-26a expression was significantly downregulated in both the kidney and the heart after Ang-II infusion. Our study identified LIMS1 as a novel target gene of miR-26a in both heart and kidney tissues. Downregulation of miR-26a activated the LIMS1/integrin-linked kinase (ILK) signaling pathway in the heart and kidney, which represents a common molecular mechanism underlying inflammation and fibrosis in heart and kidney tissues during CKD. Furthermore, knockout of miR-26a worsened inflammation and fibrosis in the heart and kidney by inhibiting the LIMS1/ILK signaling pathway; on the contrary, supplementation with exogenous miR-26a reversed all these changes. CONCLUSIONS Our findings suggest that miR-26a could be a promising therapeutic target for the treatment of cardiorenal injury in CKD. This is attributed to its ability to regulate the LIMS1/ILK signaling pathway, which represents a common molecular mechanism in both heart and kidney tissues.
Animals ; MicroRNAs/metabolism* ; Angiotensin II/toxicity* ; Mice ; Renal Insufficiency, Chronic/chemically induced* ; Mice, Knockout ; Disease Models, Animal ; Male ; Signal Transduction/genetics* ; LIM Domain Proteins/genetics* ; Mice, Inbred C57BL ; Cell Line ; Humans

BACKGROUND: Diabetic foot is a complex condition with high incidence, recurrence, mortality, and disability rates. Current treatments for diabetic foot ulcers are often insufficient. This study was conducted to identify potential therapeutic targets for diabetic foot. METHODS: Datasets related to diabetic foot and diabetic skin were retrieved from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) were identified using R software. Enrichment analysis was conducted to screen for critical gene functions and pathways. A protein interaction network was constructed to identify node genes corresponding to key proteins. The DEGs and node genes were overlapped to pinpoint target genes. Plasma and chronic ulcer samples from diabetic and non-diabetic individuals were collected. Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assays were performed to verify the S100 calcium binding protein A9 (S100A9), inflammatory cytokine, and related pathway protein levels. Hematoxylin and eosin staining was used to measure epidermal layer thickness. RESULTS: In total, 283 common DEGs and 42 node genes in diabetic foot ulcers were identified. Forty-three genes were differentially expressed in the skin of diabetic and non-diabetic individuals. The overlapping of the most significant DEGs and node genes led to the identification of S100A9 as a target gene. The S100A9 level was significantly higher in diabetic than in non-diabetic plasma (178.40 ± 44.65 ng/mL vs. 40.84 ± 18.86 ng/mL) and in chronic ulcers, and the wound healing time correlated positively with the plasma S100A9 level. The levels of inflammatory cytokines (tumor necrosis factor-α, interleukin [IL]-1, and IL-6) and related pathway proteins (phospho-extracellular signal regulated kinase [ERK], phospho-p38, phospho-p65, and p-protein kinase B [Akt]) were also elevated. The epidermal layer was notably thinner in chronic diabetic ulcers than in non-diabetic skin (24.17 ± 25.60 μm vs. 412.00 ± 181.60 μm). CONCLUSIONS S100A9 was significantly upregulated in diabetic foot and was associated with prolonged wound healing. S100A9 may impair diabetic wound healing by disrupting local inflammatory responses and skin re-epithelialization.
Calgranulin B/therapeutic use* ; Diabetic Foot/metabolism* ; Humans ; Datasets as Topic ; Computational Biology ; Mice, Inbred C57BL ; Animals ; Mice ; Protein Interaction Maps ; Immunohistochemistry

BACKGROUND: Regulatory dendritic cell (DCreg) subset exhibits a unique capacity for inducing immune tolerance among the variety subsets of dendritic cells (DCs) within gut-associated lymphoid tissues (GALTs). Fms-like tyrosine kinase 3 ligand (FLT3L) is involved in the differentiation of DCregs and the subsequent expansion of regulatory T-cells (Tregs) mediated by DCregs, though the precise mechanism remains poorly understood. This study aimed to explore the expansion mechanism of Treg induced by DCreg and the role of FLT3L in this process. METHODS: DCregs were distinguished from other DC subsets isolated from GALTs of BALB/c mice through a mixed lymphocyte reaction assay. The functions and mechanisms by which FLT3L promoted Treg expansion via DCregs were investigated in vitro through co-culture experiments involving DCregs and either CD4 + CD25 - T-cells or CD4 + CD25 + T-cells. Additionally, an in vivo experiment was conducted using a dextran sulfate sodium (DSS)-induced colitis model in mice. RESULTS: CD103 + CD11b + DC exhibited DCreg-like functionality and was identified as DCreg for subsequent investigation. Analysis of Foxp3 + Treg percentages within a co-culture system of CD4 + CD25 - T-cells and DCregs, with or without FLT3L, demonstrated the involvement of the FLT3/FLT3L axis in driving the differentiation of precursor T-cells into Foxp3 + Tregs induced by DCregs. Cell migration and co-culture assays revealed that the FLT3/FLT3L axis enhanced DCreg migration toward Tregs via the Rho pathway. Additionally, it was observed that DCregs could promote Treg proliferation through the Notch pathway, as inhibition of Notch signaling by DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) suppressed Treg expansion within the co-culture system of DCregs and CD4 + T-cells or CD4 + CD25 + T-cells. Furthermore, the FLT3/FLT3L axis influenced JAG1 expression in DCregs, indirectly modulating Treg expansion. In vivo experiments further established that FLT3L promoted DCreg expansion and restored Treg balance in DSS-induced colitis models, thereby ameliorating colitis symptoms in mice. CONCLUSION The FLT3/FLT3L axis is integral to the maintenance of DCreg function in Treg expansion.
Animals ; T-Lymphocytes, Regulatory/immunology* ; Dendritic Cells/immunology* ; Mice ; Mice, Inbred BALB C ; Membrane Proteins/metabolism* ; Receptors, Notch/metabolism* ; Lymphoid Tissue/metabolism* ; Signal Transduction/physiology* ; Coculture Techniques ; Flow Cytometry