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

OBJECTIVE: Myocardial inflammation during myocardial infarction (MI) could be inhibited by regulating arachidonic acid (AA) metabolism. Recent studies demonstrated that Sini Decoction (SND) was identified to be an effective prescription for treating heart failure (HF) caused by MI. But the anti-inflammatory mechanism of SND remained unclear. The work was designed to investigate the anti-inflammatory mechanism of SND through the AA metabolism pathway in vitro and in vivo experiments. METHODS: An inflammatory injury model of H9c2 cells was established by lipopolysaccharide (LPS)-stimulated macrophage-conditioned medium (CM). The MI model was built by the ligation of left anterior descending (LAD) branch of coronary artery in rat. Meanwhile, the rats were divided into five groups: sham group, MI group, MI + Celecoxib group, MI + low-dose SND group (SND-L) and MI + high-dose SND group (SND-H). Cardiac function, histopathological changes and serum cytokines were examined four weeks later. Western blot analysis was conducted to verify the key enzymes levels in the AA metabolic pathway, including phospholipase A2 (PLA2), cyclooxygenases (COXs) and lipoxygenases (LOXs). RESULTS: These in vivo results demonstrated that SND could improve the cardiac function and pathological changes of rats with MI, and regulate the key inflammatory molecules in the AA metabolism pathway, including sPLA2, COX-1, COX-2, 5-LOX and 15-LOX. In vitro, SND could decrease the release of pro-inflammatory cytokines including TNF-α and IL-6 and inhibit cell apoptosis in CM-induced H9c2 cells. Moreover, SND could protect H9c2 cells from the damage of CM by regulating nuclear factor kappa-B (NF-κB) signal pathway and the expression of COX-2. CONCLUSION SND may be a drug candidate for anti-inflammatory treatment during MI by regulating the multiple targets in the AA metabolism pathway.

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective: To explore the moderating role of estradiol in the relationship between parenting styles and preschool children's behavioral problems, so as to provide a theoretical basis for improving the development of human s emotional health development in early life stage. Methods: During September to November in 2022, 354 children aged 3-6 years and their parents from two kindergartens in Bengbu City were chosen by using stratified cluster sampling method for the questionnaire survey. The Parenting Style Scale and the Child Behavior Checklist (CBCL) were used to collect information on parenting style and child behavioral problems. Salivary estradiol of children was collected and tested. Independent samples t test was applied to compare the scores of the scale for parental up bringing and children s behavioral problems, and Pearson correlation analysis was conducted to explore the relationship among parental upbringing, estradiol and children s behavioral problems. Results: Parents doting, laissez faire, autocratic, and inconsistent parenting styles were positive associated with child behavior problems( r =0.14-0.70); fathers democratic parenting style was negatively associated with child behavior problems( r =-0.14，-0.22，-0.21，-0.17，-0.27，-0.20); mothers democratic parenting styles was negatively correlated with scores on all five dimensions of child behavior problems except the withdrawal dimension ( r =-0.14，-0.12，-0.13，-0.21，-0.12)( P <0.05). Estradiol levels had significant moderating effects on maternal doting parenting style and children s withdrawal ( β =0.68) as well as social problems ( β =-1.00), also moderating laissez faire parenting styles and children s withdrawal problems ( β =0.75)( P <0.05). For children with low levels of estradiol, withdrawal problem scores were negatively associated with mother s doting parenting style and positively associated with laissez faire parenting style, and socialization problem scores were associated with mother s doting parenting style; for children with high levels of estradiol, withdrawal problem scores were positively associated with mother s doting parenting style, and socialization problem scores were associated with mother s doting parenting style ( t=2.84, 6.24, 3.16 , 2.37, 4.49, P <0.05). Conclusions Parenting styles are strongly associated with child behavioral problems; estradiol levels play a moderating role in mothers doting, laissez faire parenting styles and children s withdrawal problems and social problems.Parents should adopt more positive parenting styles and focus on the role of estradiol levels in maternal education to reduce the occurrence of behavioral problems in children.

Objective:To explore the technical points and efficacy of gradual traction-unlocking closed reduction technique (GT-UCRT) for Tile C old pelvic fractures.Methods:From August 2012 to June 2021, 6 patients with Tile C old pelvic fracture were treated and followed up at Department of Orthopedics, The Fourth Medical Center, General Hospital of Chinese PLA. They were 4 males and 2 females with an age of (35.8±10.5) years. By Tile classification: 4 cases of type C1.2, 1 case of type C1.3, and 1 case of type C2; time from injury to surgery: 153.8 (64.3, 204.8) days. The 6 patients were treated with GT-UCRT. The time for gradual traction reduction, operation time, hospital stay, intraoperative blood loss and complications were recorded. The modified Matta score was used to evaluate the reduction quality of pelvic fractures, and the Majeed score was used to evaluate the pelvic function at the last follow-up.Results:The 6 patients were followed up for (40.3±22.9) months (from 12 to 72 months) after surgery. The time for gradual traction reduction was (26.7±4.6) days, operation time (119.2±4.6) minutes, hospital stay (11.5±2.9) days, and intraoperative blood loss (533.3±189.6) mL. By the modified Matta score, the pelvic reduction after surgery was rated as satisfactory in 5 cases and as unsatisfactory in 1 case. The length disparity between both lower limbs in the 6 patients was (6.9±1.6) cm before surgery and (1.0±0.4) cm immediately after surgery, showing a statistically significant difference ( t=11.135, P<0.001). One fracture healed 3 months after surgery and 5 fractures 6 months after surgery. The Majeed pelvic score at the last follow-up was (80.8±9.0) points for the 6 patients, yielding 2 excellent cases, 3 good cases and 1 fair case. Delayed wound healing was reported in 1 patient, calf intermuscular venous thrombosis in 2 cases, and emotional anxiety and sleep disorder in 1 patient. No new lumbosacral plexus injury was found in any patient. Conclusions:In the treatment of Tile C old pelvic fractures, since our self-designed GT-UCRT combines the advantages of Ilizarov technique and unlocking closed reduction technique, it can not only protect the lumbosacral plexus but also obtain satisfactory reduction of the fracture.

Objective To investigate the molecular mechanism of taurine regulating the polarization of M2 macrophages by mitophagy. Methods THP-1 cells were divided into four groups: M0 group (THP-1 cells were treated by 100 nmol/L phorbol myristate ester for 48 hours to polarize into M0), M2 group (THP-1 cells were induced to polarize into M2 macrophages by 20 ng/mL interferon-4 (IL-4) for 48 hours), M2 combined with taurine groups (added with 40 or 80 mmol/L taurine on the basis of M2 macrophages). The mRNA expression of mannose receptor C type 1(MRC-1), C-C motif chemokine ligand 22(CCL22) and dendritic cell-specific ICAM-3 grabbing non-integrin (CD209) in M2 macrophages were detected by quantitative real-time PCR. Mitochondrial and lysosome probes were used to detect the number of mitochondria and lysosomes by multifunction microplate reader and confocal laser scanning microscope. The level of mitochondrial membrane potential (MMP) was detected by JC-1 MMP assay kit. The expression of mitophagy-related proteins PTEN-induced putative kinase 1 (PINK1) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blot analysis. Results Compared with M0 group, the expression of MRC-1, CCL22, CD209 and PINK1, the number of mitochondria and the level of MMP in M2 group were significantly increased, whereas the number of lysosomes and LC3II/LC3I ratio were decreased. Compared with M2 group, the expressions of MRC-1, CCL22 and CD209, the number of mitochondria and the level of MMP in M2 combined with taurine group dropped significantly while the number of lysosomes was found increased, and the protein expression of PINK1 and LC3II/LC3I ratio were also increased. Conclusions The polarization of M2 macrophages is regulated by taurine to prevent excessive polarization via reducing the level of MMP, improving the level of mitophagy, reducing the number of mitochondria, and inhibiting the mRNA expression of polarization markers in M2 macrophages.
Mitophagy ; Taurine ; Macrophages/metabolism* ; Protein Kinases/metabolism* ; RNA, Messenger

Objective:To analyze clinical features and prognosis of Mycoplasma pneumoniae-induced rash and mucositis (MIRM) . Methods:Among patients who were diagnosed with erythema multiforme/severe erythema multiforme or Stevens-Johnson syndrome at discharge from the First Affiliated Hospital, Sun Yat-sen University from November 2004 to May 2021, patients with MIRM were screened out according to diagnostic criteria for MIRM and after exclusion of other causes, and their clinical manifestations, laboratory and auxiliary examinations, treatment and prognosis were analyzed.Results:Eight patients were found to meet the MIRM diagnostic criteria, including 4 males and 4 females, with the age at onset being 15.63 ± 9.16 years (range, 4 - 30 years) . All the 8 patients had fever, and 5 of them had upper respiratory symptoms such as cough and sore throat. Oral mucosal damage occurred in all the patients, 5 of whom presented with blood crusts on the lips; eye damage occurred in 7 patients, which manifested as conjunctiva hyperemia and increased secretions. All the patients presented with skin lesions, including 5 with targetoid lesions and 4 with blisters. All the patients were serologically positive for anti- Mycoplasma pneumoniae IgM. One patient experienced recurrent upper respiratory tract infections such as dry cough, each episode was closely related to Mycoplasma pneumoniae infection, and whole exome sequencing of the peripheral blood showed heterozygous mutations in the NLRC4 and IRGM genes. Histopathological examination of skin lesions was performed in 3 patients, and the results were consistent with the diagnosis of erythema multiforme. Seven patients were treated with systemic glucocorticoids, 6 with intravenous immunoglobulin, 5 with azithromycin, and 5 with acyclovir, valacyclovir or ribavirin. After an average 2.9-year follow-up, 3 patients were cured, 1 was blind, 1 experienced recurrent dry cough, oral ulcers and rashes on the limbs, and the remaining 3 developed eye damage such as meibomian gland dysfunction, punctal stenosis and corneal epithelial damage. Conclusions:MIRM mostly occurred in children and young adults, and was mainly accompanied by prodromal symptoms such as fever, sore throat and cough. MIRM mainly manifested as obvious mucosal damage and some targetoid lesions. Most patients could recover after a single attack, and recurrent episodes may be related to mutations in autoinflammation- and infection-related genes in some patients.

Objective:To evaluate the clinical efficacy of cannulated screws with sutures in the treatment of patellar transverse fractures.Methods:A retrospective analysis was performed of the data of 70 patients with patellar transverse fracture who had been admitted to Senior Department of Orthopedics, The Fourth Medical Center of Chinese PLA General Hospital from January 2017 to March 2021. According to the construction methods for the tension band, the patients were divided into 3 groups. In group A of 21 cases subjected to fixation with cannulated screws with sutures (Fiber-Tape), there were 8 males and 13 females with a median age of 55.0 (48.0, 65.0) years; in group B of 32 cases subjected to fixation with Kirschner wire tension band, there were 15 males and 17 females with a median age of 52.5 (41.5, 63.0) years; in group C of 17 cases subjected to fixation with Cable-Pin system, there were 5 males and 12 females with a median age of 55.0 (37.0, 65.0) years. The 3 groups were compared in terms of complications, secondary surgery (removal of internal fixation), operation time, intraoperative blood loss and knee function rated by the Lysholm and B?stman scores at the last follow-up.Results:There were no significant differences in the preoperative general data between the 3 groups, showing they were comparable ( P > 0.05). There was no significant difference in the operation time, intraoperative blood transfusion or follow-up time among the 3 groups ( P > 0.05). The incidence of soft tissue irritation [4.8% (1/21)] and the secondary operation rate [4.8% (1/21)] in group A were significantly lower than those in group B [43.8% (14/32) and 37.5% (12/32)] and group C [41.2% (7/17) and 35.3% (6/17)] ( P < 0.05), but there was no statistically significant difference between group B and group C ( P > 0.05). In groups A, B and C, respectively, the Lysholm knee score was 84.0 (69.0, 88.0), 89.0 (71.5, 95.0) and 82.0 (63.0, 90.0), and the B?stman knee score 26.0 (23.0, 28.0), 26.5 (24.0, 27.5) and 26.0 (22.0, 28.0), showing no significant difference ( P > 0.05). There was no significant difference either in the incidence of other complications among the 3 groups ( P > 0.05). Conclusion:In the treatment of patellar transverse fractures, compared with the Kirschner wire tension band and Cable-Pin system, cannulated screws with sutures (Fiber-Tape) may lead to a lower incidence of soft tissue irritation and a lower rate of secondary surgery, but no significant differences in operation time, intraoperative blood loss, other complications or postoperative functional scores.

BACKGROUND: With the extensive development of minimally invasive surgery for pulmonary nodules, preoperative localization becomes more and more critical. There are some defects in traditional localization methods, so it is necessary to improve. The aim of this study was to compare and analyze the safety and effectiveness of two new methods, namely four-hook needle and memory alloy coil, in the localization of pulmonary nodules. METHODS: A retrospective analysis of 152 patients was performed. 76 cases were in four-hook needle group, and 76 cases were in memory alloy coil group. Pulmonary nodules were located before operation, and then video-assisted wedge resection was performed. The average procedure time, localization complications and nodule resection time were counted. RESULTS: The target pulmonary nodules were successfully removed in both groups. In four-hook needle group, 76 patients found localization devices, all the pulmonary nodules were successfully removed, and one case was transferred to open the chest for wedge resection of pulmonary nodules due to severe thoracic adhesion. All 76 patients in memory alloy coil group were successfully resected with pulmonary nodules, and one patient underwent compromising enlarged resection because no lesion was found after the specimen was removed during the operation. There was no significant difference in the incidence of pneumothorax and pulmonary hemorrhage, the success rate of localization and nodule wedge resection time between the two groups. The average time of localization in four-hook needle group was (13.66±3.11) min, lower than that of memory alloy coil group (15.51±3.65) min, and the difference was statistically significant (P=0.001). In memory alloy coil group, when the distance from the nodule to the pleura was ≥1.5 cm and <1.5 cm, the average localization time was (17.20±4.46) min and (14.91±3.15) min, respectively, and there was a statistical difference between the two distance (P=0.044). CONCLUSIONS Four-hook needle and memory alloy coil have good safety and effectiveness, and the localization time of four-hook needle is shorter. When using memory alloy coil, the effect of the method is better for pulmonary nodules with a distance less than 1.5 cm to pleura.
Alloys ; Humans ; Lung Neoplasms/surgery* ; Multiple Pulmonary Nodules ; Retrospective Studies ; Thoracic Surgery, Video-Assisted ; Tomography, X-Ray Computed

Objective: To investigate the relationship between SUVmax on preoperative 68Ga-PSMA PET-CT and the clinicopathological characteristics of patients treated with radical prostatectomy. Methods: The clinicopahtological data of patients evaluated with ⁶⁸Ga-PSMA PET-CT preoperatively and treated with radical prostatectomy between May 2016 and August 2019 were retrospectively reviewed. 31 patients with a mean age (63.1±4.9) and baseline PSA (72.71±173.15)ng/ml were enrolled. Their BMI mean (24.6±3.0)kg/m². Baseline testosterone of 14 patients was (4.72±1.64)ng/ml.Based on the Gleason scores related ISUP classification, all patients were classified into grade one in 5 cases, grade 2in 7 cases, grade 3 in 4 cases, grade 4 in 10 cases and grade 5 in 5 cases. The clinical classification included 6 cases in T_2a stage, 2 cases in T_2b stage, 17 cases in T_2c stage, 1 case in T_3a stage, 4 cases in T_3b stage and 1 case in T₄ stage. SUVmax was accessed by two independent professional nuclear medicine physicians. SUVmax was 12.49±9.38. SPSS 16.0 software was used to do statistic analysis. Results: The post-operative pathological results showed the surgical margin positive in 19 cases, negative in 12 cases, vascular positive in 5 cases, negative in 20 case, positive nerve invasion in 20 cases and negative in 11 cases. 2 patients were low risk, 7 patients were medium risk and 22 patients were high risk according to D′Amico classification. Based on the basis of PSA(≤10 or>10) and Gleason score(≤6 or>6), 6 patients were in group with low PSA and low Gleason score, 5 patients were low PSA and high Gleason score, 9 patients were high PSA and low Gleason score, 11 patients were high PSA and high Gleason score. SUVmax had a significant positive relationship with pathological ISUP(r=0.434, P=0.015) and SUVmax in patients with positive intravascular tumor emboli was significantly higher than those with negative intravascular tumor emboli(14.78±10.68 vs. 8.17±2.81, P=0.005). No significant correlation was found between SUVmax and baseline PSA, testosterone, pathologic T stage, surgical margin, nerve invasion, pelvic lymph node status as well as risk stratification. SUVmax could distinguish pathologic ISUP grade 5 with a maximum AUC 0.747 (P=0.033) and the sensitivity was 88.9%. The specificity was 77.3% when SUVmax≥11.34. SUVmax in patients with upgrading ISUP was significantly higher than that in patients with downgrading ISUP (16.01±5.40 vs. 4.98±2.11, P=0.007). Conclusions SUVmax measured on preoperative ⁶⁸Ga-PSMA PET-CT may have a clinical significance in predicting unfavorable pathological factors for patients treated with radical prostatectomy.