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

Background: The function of CD69 expressed on T cells in chronic hepatitis B (CHB) remains unclear. We aimed to elucidate the roles of CD69 on T cells in the disease process and in antiviral therapy for CHB. Methods: We enrolled 335 treatment-naive patients with CHB and 93 patients with CHB on antiviral therapy. CD69, antiviral cytokine production by T cells, T-helper (Th) cells, and inhibitory molecules of T cells were measured using flow cytometry, and clinical-virological characteristics were examined dynamically during antiviral therapy. Results: CD69 expression on CD3+, CD4+, and CD8+ T cells was the lowest in the immune-active phase and was negatively correlated with liver transaminase activity, fibrosis features, inflammatory cytokine production by T cells, and Th-cell frequencies but positively with inhibitory molecules on T cells. CD69 expression on CD3+, CD4+, and CD8+ T cells decreased after 48 weeks of antiviral therapy, and patients with hepatitis B e antigen (HBeAg) seroconversion in week 48 showed lower CD69 expression on T cells at baseline and week 48. The area under the ROC curve of CD69 expression on T cells at baseline for predicting HBeAg seroconversion in week 48 was 0.870, the sensitivity was 0.909, and the specificity was 0.714 (P = 0.002). Conclusions CD69 negatively regulates T-cell immunity during CHB, and its expression decreases with antiviral therapy. CD69 expression predicts HBeAg seroconversion in week 48. CD69 may play an important negative role in regulating T cells and affect the efficacy of antiviral therapy.

Objective:To study the osseointegration and new bone formation of titanium implants treated by sandblasting acid etching combined with plasma spraying tantalum coating in rats.Methods:After polishing and cleaning of the pure titanium specimens(SLA),the tantalum coating was prepared by plasma spraying after sandblasting and acid etching(SLA-Ta).The surface feature of the samples was observed by SEM,EDS and contact angle meter.The samples of SLA and SLA-Ta were respectively implanted into femur condyle at each side of 18 SD rats(SLA group and SLA-Ta group),and the rats were double-fluorescent labeled after surgery.The rats were sacrificed 4 and 12 weeks after implantation respectively(n=9),the samples of implants and surrounding tissue were examined by mi-cro-CT scanning,fluorescence microscopy and histological staining,the new bone formation and osseointegration around the implant were analyzed.Results:Micro-nano Porous structureed tantalum coating was successfully prepared on the surface of SLA titanium.In vivo experiments showed that 4 and 12 weeks after implantation,the amount of new bone and osseointegration around the implant in the SLA-Ta group were significantly higher than those in the SLA group.Conclusion:The micro-nano multi-level structure formed by SLA combined with plasma sprayed tantalum can promote new bone formation,accelerate mineralization deposition,and improve the osseoin-tegration capacity of the implant.

Objective:To explore the effect of timely induction intervention on postpartum urination in primipara during vaginal delivery, so as to provide the evidence for preventing the occurrence of postpartum urinary retention and relieving the pain of primipara.Methods:This study adopted a randomized controlled trial design, and selected 400 cases of primipara who were hospitalized for vaginal delivery in the Obstetric Department of Dalian Women and Children's Medical Group Sports New Town Hospital from June 2021 to September 2022 as the study objects by convenience sampling method. They were divided into the intervention group and the control group with 200 cases each by random number table method, and the control group received routine postpartum care. Instruct active urination within 6 hours after delivery. The intervention received timely induction urination intervention. The general condition and bladder urine volume of the women in the intervention group were evaluated at 2, 4, 6 h after delivery, respectively, and personalized guidance was implemented, including the frequency of massage of the bottom of the uterus, the control of water intake, the selection of methods and timing of inducing urination, etc., and routine postpartum care was given when the women completed their first urination and had no complaints of discomfort. The first urination time, first urination volume, first bladder irritation during the first urination and the incidence of postpartum urinary retention in different periods were compared between the two groups.Results:The patients in the control group were (29.60 ± 3.20) years old, while the patients in the intervention group were (28.81 ± 3.42) years old. The first urination time in the intervention group was (6.89 ± 2.18) h, which was shorter than that in the control group (9.11 ± 3.86) h, and the difference was statistically significant ( t=-2.49, P<0.01). The first urination volume in the intervention group was (322.36 ± 120.15) ml, which was higher than that in the control group (262.93 ± 105.68) ml, and the difference was statistically significant ( t=3.39, P<0.05). The incidence of the first bladder irritation in the intervention group was 22.0%(44/200), which was lower than that in the control group 33.5%(67/200), and the difference was statistically significant ( χ2=6.60, P<0.05). The incidence of postpartum urinary retention within 24 h in the intervention group was 5.5%(11/200), which was lower than that in the control group 11.5%(23/200), and the difference was statistically significant ( χ2=4.63, P<0.05). The incidence of postpartum urinary retention within 1 week in the intervention group was 9.5%(19/200), which was lower than that in the control group 16.5%(33/200), and the difference was statistically significant ( χ2=4.33, P<0.05). There was no significant difference in the incidence of postpartum urinary retention within 24 to 72 h between the two groups ( P>0.05). Conclusions:Timely induction intervention can reduce the incidence of postpartum urinary retention, shorten the time of first urination, increase the volume of first urination and improve the comfort of first urination, which is worthy of clinical application.

Objective To investigate the protective effects of Sulforaphane(SFN)against high glucose(HG)-induced human platelet apoptosis and elucidate the underlying mechanisms in vitro.Methods Purified platelets obtained from healthy individuals were pre-incubated with various concentrations of SFN(5,10,or 20 μmol/L)or a vehicle control(0.05%DMSO)for 40 minutes at 37℃.Subsequently,the platelets were stimulated with normal glucose(NG,5 mmol/L)or high glucose(HG,25 mmol/L)for an additional duration of 90 minutes.Flow cytometry was employed to assess platelet mitochondrial membrane potential depolarization(Δψm),exposure of phosphatidylserine(PS),and generation of total intraplatelet reactive oxygen species(ROS).Phosphorylation levels of PI3K and Akt were determined using Western blot.Results Compared to NG-treated human platelets,HG significantly induced depolarization of platelet Δψm and exposure of PS(P＜0.001).These effects of HG were markedly attenuated by various concentrations of SFN(P＜0.001).Mechanistically,SFN down-regulated the phosphorylation levels of PI3K(P＜0.01)and Akt(P＜0.05),which were increased by HG when compared to the vehicle control,and substantially reduced total intracellular ROS levels(P＜0.001).The inhibitory effects of SFN on HG-induced phosphorylation of PI3K and Akt,as well as its efficacy on total intracellular ROS generation,Δψm depolarization,and PS exposure in HG-stimulated human platelets were completely reversed by a specific agonist for PI3K,740 Y-P.Conclusions The current study demonstrates that SFN exerts a protective effect on platelet apoptosis induced by HG,potentially through the down-regulation of the PI3K/Akt-mediated pathway in human platelets in vitro.These findings suggest that SFN may hold promise for improving thrombosis in diabetes mellitus and related chronic metabolic diseases.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is characterized by diffuse alveolar injury primarily caused by an excessive inflammatory response. Regrettably, the lack of effective pharmacotherapy currently available contributes to the high mortality rate in patients with this condition. Xuebijing (XBJ), a traditional Chinese medicine recognized for its potent anti-inflammatory properties, exhibits promise as a potential therapeutic agent for ALI/ARDS. This study aimed to explore the preventive effects of XBJ on ALI and its underlying mechanism. To this end, we established an LPS-induced ALI model and treated ALI mice with XBJ. Our results demonstrated that pre-treatment with XBJ significantly alleviated lung inflammation and increased the survival rate of ALI mice by 37.5%. Moreover, XBJ substantially suppressed the production of TNF-α, IL-6, and IL-1β in the lung tissue. Subsequently, we performed a network pharmacology analysis and identified identified 109 potential target genes of XBJ that were mainly involved in multiple signaling pathways related to programmed cell death and anti-inflammatory responses. Furthermore, we found that XBJ exerted its inhibitory effect on gasdermin-E-mediated pyroptosis of lung cells by suppressing TNF-α production. Therefore, this study not only establishes the preventive efficacy of XBJ in ALI but also reveals its role in protecting alveolar epithelial cells against gasdermin-E-mediated pyroptosis by reducing TNF-α release.
Animals ; Mice ; Alveolar Epithelial Cells ; Pyroptosis ; Gasdermins ; Lipopolysaccharides/adverse effects* ; Tumor Necrosis Factor-alpha ; Acute Lung Injury/drug therapy* ; Respiratory Distress Syndrome

Objective:To discuss the clinical features, diagnosis and treatment of linear scleroderma (LS).Methods:A case of LS diagnosed in the Second Hospital of Shandong University in October 20, 2020, was reported and the clinical features and pathological documentation of the disease reported in the literature were reviewed.Results:A 24-year-old woman presented cicatricial alopecia on the left frontoparietal area and facial atrophy for about 10 years. Two years before, she began to suffer ptosis and neurological complaints. Clinical features of different stages of the disease are presented. All 15 patients reported in the literature were analyzed, with a median of 22 years and a male to female ratio of 9∶6. There were 4 cases of linear scleroderma with ipsilateral drooping eyelids and lateral contraction, 3 cases of linear scleroderma with demyelinating lesions, combined with lateral contraction, 3 cases of linear scleroderma combined with lateral atrophy, and 1 case of linear scleroderma with ipsilateral facial spasm. Two cases were with the chest sclerosing spot. Two cases of linear scleroderma were with epileptic seizure and white matter demyelination lesion. Six cases were treated with hormone, 2 cases were treated with methotrexate. One case was treated with both hormone and methotrexate. One case was treated with botulinum toxin. Three cases were treated with surgical correction of eyelid ptosis. One case was treated with ultraviolet A1 radiation phototherapy and 1 case was treated with vitamin therapy.Conclusions:Patients with scleroderma may have ipsilateral facial atrophy, blepharoptosis and facial spasm. Some patients involving the nervous system may have epilepsy and myelitis. And demyelinating lesions can be seen in magnetic resonance imaging. Localized scleroderma may develop into systemic scleroderma. Therefore, it is recommended to combine immunosuppressants as soon as possible to control the development of the disease if necessary.

OBJECTIVE：To e stablish the method for simultaneous determination of 10 kinds of active components in Tibetan medicine Siwei jianghuang prescription ，and to optimize its decoction technology. METHODS ：HPLC method was adopted. Using soaking time ，the amount of added water ，decoction time and decoction times as factors ，comprehensive score of the contents of 10 kinds of components and solid extracts rate as response values ，one the basis of single factor test ，Box-Behnken response surface method was used to optimize its decoction technology. RESULTS ：The linear range of gallic acid ，corilagin，magnoflorine， ellagic acid ， hydrochloric jatrorrhizine ， hydrochloride palmatine ， hydrochloride berberine ， bisdemethoxycurcumin， demethoxycurcumin and curcumin were 0.280 6-1.683 6，0.289 6-1.737 6，0.320 8-1.924 8，0.116 0-0.696 0，0.018 9-0.113 5， 0.013 3-0.079 9，0.092 3-0.553 8，0.025 5-0.153 0，0.036 1-0.216 3，0.041 0-0.245 7 µg（all r were 0.999 9），respectively. The limits of quantitation were 0.28，14.48，3.21，11.60，1.89，4.44，0.46，0.26，0.36，0.41 ng，respectively. The limits of detection were 0.11，4.14，1.24，3.32，0.58，1.33，0.13，0.09，0.14，0.12 ng，respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％. The recoveries were 92.56％-103.69％（RSDs were 0.90％-3.81％，n＝6）. The optimal decoction technology included soaking 60 min，adding 8-fold（mL/g）water，decoction for twice ，lasting for 65 min each time. In 6 validation tests ，comprehensive scores were 3.323 2-3.422 4，and the absolute value of the relative error with the predicted value （3.437 4）was less than 2％.CONCLUSIONS：Established method is simple and repeatable ，and can be used for simultaneous determination of 10 kinds of active components in Siwei jianghuang prescription. Optimized decoction technology is stable and feasible.

Biochemistry is an important fundamental course of biology related majors, and has the characteristics of speedy development, massive information, sound theoretical basis and feasible applicability. It is difficult for students to learn well in the process of teaching. In addition, the experimental course lacks integrity, comprehensiveness and design experiments. Under the guidance of outcome-based education (OBE) concept, we established a multi-dimensional teaching reform system through theoretically and practically introducing of various teaching methods, online courses, bilingual teaching, stronger practical teaching and optimized assessment mode. The teaching reform system could effectively allow students to change from "passive learning" to "active learning" through activating learning enthusiasm, resulting in cultivated innovation ability. This system could play very important role in promoting quality of talent training of colleges and universities.
Biochemistry/education* ; China ; Humans ; Learning ; Students ; Universities/trends*