OBJECTIVES: To fabricate an injectable composite microsphere hydrogel reinforced with silk fibroin/hyaluronic acid microspheres, achieving synergistic enhance-ment of mechanical robustness and biofunctionality. METHODS: Methacrylated hyaluronic acid (HAMA) and thiolated silk fibroin (TSF) were synthesized. Monodisperse microspheres generated via microfluidics were UV-cured (420 nm) through thiol-ene click reaction. These microspheres were embedded in a TSF/HAMA matrix to form photo-cured composites. The grafting rate of TSF and HAMA was characterized by H1-NMR; particle size distribution of microsphere hydrogels in soybean oil was observed by optical microscopy; gel point of composite microsphere hydrogels was determined by advanced extensional rheometer; microscopic morphology of microsphere hydrogels was observed by scanning electron microscopy; elemental distribution of microsphere hydrogels was detected by X-ray energy dispersive spectroscopy; tunability of composite microsphere hydrogels was observed by inverted confocal microscopy; mechanical properties of composite microsphere hydrogels were tested by compression testing; swelling ratio, degradation rate and water retention rate of composite microsphere hydrogels were measured by gravimetric method. Cytotoxicity of the composite microsphere hydrogels was determined by Calcein-AM/propidium iodide dual staining and CCK-8 assay; cell migration capability was observed by scratch assay. RESULTS: The grafting rates of HAMA and TSF was 48.03% and 17.99%, respectively. Microsphere hydrogels with particle sizes of (43.3±1.2), (78.1±3.0), and (130.8±1.9) μm were prepared. The gel time of the composite microsphere hydrogels was 48-115s. The laser confocal imaging confirmed dynamic regulation characteristics of the composite microsphere hydrogels. The compressive strength of the composite microsphere hydrogels reached 22.7 kPa and maintained structural integrity at 40% strain after 20 compression cycles. The composite microsphere hydrogels exhibited differential deswelling behaviors in simulated physiological environments, and reducing microsphere particle size could significantly enhance its stability under moist conditions. The degradation rate of the composite microsphere hydrogels was (49.1±0.9)% after 200 h, and water retention rate was maintained at 40%-60% after 96 h. Biocompatibility assays confirmed >95% cell viability and unimpaired cell migration abilities. CONCLUSIONS The TSF/HAMA composite microsphere hydrogel developed in this study has characteristics of rapid fabrication, adjustable mechanical properties, enhanced environmental stability and excellent biocom-patibility, thus providing a new material solution for tissue repair and regenerative medicine.
Fibroins/chemistry* ; Hydrogels/chemistry* ; Microspheres ; Hyaluronic Acid/chemistry* ; Humans

Sennoside A (SA), a typical prodrug, exerts its laxative effect only after its transformation into rheinanthrone catalyzed by gut microbial hydrolases and reductases. Hydrolases have been identified, but reductases remain unknown. By linking a photoreactive group to the SA scaffold, we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling (ABPP). From lysates of an active strain, Bifidobacterium pseudocatenulatum (B. pseudocatenulatum), 397 proteins were enriched and subsequently identified using mass spectrometry (MS). Among these proteins, chromate reductase/nicotinamide adenine dinucleotide (NADH) phosphate (NADPH)-dependent flavin mononucleotide (FMN) reductase/oxygen-insensitive NADPH nitroreductase (nfrA) was identified as a potent SA reductase through further bioinformatic analysis and The Universal Protein Resource (UniProt) database screening. We also determined that recombinant nfrA could reduce SA. Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.

Objective To investigate the expression of Toll-like receptor 2(TLR2)and Toll-like receptor 4(TLR4)in peripheral blood mononuclear cells(PBMC)in children with recurrent respiratory tract infection(RRTI)and its relationship with T helper cell 1(Th1)/T helper cell 2(Th2)immune response.Methods A total of 65 children diagnosed with RRTI who admitted to the hospital from December 2020 to December 2022 were enrolled in the study as the RRTI group,and 45 healthy children who underwent physical examination in the hospital during the same period were enrolled as the control group.The relative expression levels of TLR2 and TLR4 mRNA in PBMCs were detected by real-time fluorescence quantitative PCR(qPCR).The expres-sion rates of TLR2 and TLR4 protein in PBMCs were detected by flow cytometry.The levels of Th1 cytokine interferon-γ(IFN-γ),Th2 cytokine interleukin-4(IL-4)and their ratio(IFN-γ/IL-4)in plasma were detected by enzyme-linked immunosorbent assay(ELISA).Pearson correlation analysis was used to analyze the corre-lation between TLR2,TLR4 protein expression rates and plasma IFN-γ,IL-4 levels.Results The RRTI group had significantly higher plasma level of Th2 cytokine IL-4 than the control group,significantly lower plasma level of Th1 cytokine IFN-y than the control group,and significantly lower ratio of IFN-γ/IL-4 than the con-trol group,the differences were all statistically significant(P＜0.05).The relative expression levels of TLR2 and TLR4 mRNA and protein expression rates in PBMC of children in the RRTI group were higher than those in the control group,and the differences were statistically significant(P＜0.05).Pearson correlation analysis showed that the protein expression rates of TLR2 and TLR4 in PBMC of children with RRTI were both nega-tively correlated with both plasma IFN-γ levels and IFN-γ/IL-4(P＜0.05)and positively correlated with plasma IL-4 levels(P＜0.05).Conclusion The expression of TLR2 and TLR4 in PBMC and plasma Th1/Th2 cytokines in children with RRTI may be involved in the occurrence and development of the disease.Ex-cessive activation of TLR2 and TLR4 may weaken Th1 function and enhance Th2 function.

Objective:Laboratory evaluation of the relationship between cerebrospinal fluid and plasma indicators and intrathecal immunoglobulin G(IgG) synthesis in patients with neurological diseases, and establishment of a new diagnostic method for intrathecal IgG synthesis.Methods:This study retrospectively analyzed the content of IgG in cerebrospinal fluid samples and blood albumin in blood samples, and other test results of 410 patients with neurological diseases who visited Beijing Tiantan Hospital from 2019 to 2022. According to the results of oligoclonal bands in cerebrospinal fluid, patients were divided into intrathecal IgG synthesis group and non-intrathecal IgG synthesis group. The Mann Whitney U test was used for inter group comparison, and a bilateral test with P<0.05 indicates a statistically significant difference. Include indicators with differences between groups in logistic regression analysis, construct a predictive model, and compare it with the established quantitative formula IgG index. Results:There were significant differences in 10 indicators, including cerebrospinal fluid leukocyte count and 24-hour intrathecal IgG synthesis rate, between the intrathecal IgG synthesis group and the non-intrathecal IgG synthesis group, with P<0.05. The area under the curve (AUC) of intrathecal IgG synthesis was higher than the IgG index (AUC=0.920, 0.809, Z=31.178, P<0.001), the sensitivity was higher than the IgG index (0.825, 0.618), and the specificity was lower than the IgG index (0.876, 0.908). Conclusion:The combination of 10 indicators such as cerebrospinal fluid white blood cell count and 24-hour intrathecal IgG synthesis rate can improve the diagnostic efficacy and sensitivity of intrathecal IgG synthesis.

Objective: To explore and validate the potential targets of Paeoniae Radix Alba (P. Radix, Bai Shao) in protecting against chemical liver injury through network pharmacology, molecular docking technology, and in vitro cell experiments. Methods: Network pharmacology was used to identify the common potential targets of P. Radix and chemical liver injury. Molecular docking was used to fit the components, which were subsequently verified in vitro. A cell model of hepatic fibrosis was established by activating hepatic stellate cell (HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1. The cells were exposed to different concentrations of total glucosides of paeony (TGP), the active substance of P. Radix, and then evaluated using the cell counting kit-8 assay, enzyme-linked immunosorbent assay, and western blot. Results: Analysis through network pharmacology revealed 13 key compounds of P. Radix, and the potential targets for preventing chemical liver injury were IL-6, AKT serine/threonine kinase 1, jun proto-oncogene, heat shock protein 90 alpha family class A member 1 (HSP90AA1), peroxisome proliferator activated receptor gamma (PPARG), PTGS2, and CASP3. Gene Ontology (GO) enrichment analysis indicated the involvement of response to drugs, membrane rafts, and peptide binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation. Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1, PTGS2, PPARG, and CASP3. Different concentrations of TGP can inhibit the expression of COL-Ⅰ, COL-Ⅲ, IL-6, TNF-α, IL-1β, HSP-90α, and PTGS2 while increasing the expression of PPAR-γ and CASP3 in activated HSC-LX2 cells. Conclusion P. Radix primarily can regulate targets such as HSP90AA1, PTGS2, PPARG, CASP3. TGP, the main active compound of P. Radix, protects against chemical liver injury by reducing the inflammatory response, activating apoptotic proteins, and promoting the apoptosis of activated HSCs.

Purpose To investigate the clinical,imaging,pathological,and genetic characteristics of neuroepithelial tumors with EWSR1 translocation.Methods The clinicopatho-logical data of 6 patients with EWSR1 translocation in neuroepi-thelial tumors were collected,routine HE and immunohistochem-ical staining were performed,the information of high-throughput sequencing was summarized,and the relevant literature was re-viewed.Results The median age of the 6 patients was 11.5 years(ranging from 1.9 to 17 years),including 1 male and 5 females.The tumors located in temporal lobe,frontal lobe,pari-etal lobe,suprasellar region,or lateral ventricle.The clinical manifestations mainly started with seizures.Brain MRI showed abnormal signal focus in the cerebral hemisphere near the cortex in 4 cases,and ventricle/periventricular regions in 2 cases,with an almost clear boundary in 5 cases.Microscopically,the histo-logical changes were diverse,including low-grade gliomas/gli-oneuronal tumors in 3 cases,high-grade gliomas in 2 cases,and glioneuronal tumor with high-grade feature in 1 case.Immuno-histochemically,tumor cells expressed GFAP,S-100,Syn,and Olig2 partially.2 cases exhibited slightly positive of NeuN and 1 case exhibited little dot-like staining of EMA.Next generation sequencing revealed EWSR1 rearrangement in all 6 cases,with chaperone genes including PATZ1 in 5 cases,and PLAGL1 in 1 case.3 cases were treated with chemotherapy after surgery,and no recurrence or progression was found during follow-up.Con-clusion The neuroepithelial tumors with the fusion of EWSR1 and non-ETS commonly occur in the cerebral hemisphere of teenagers and children.Most of the boundaries lesion are still clear,the histomorphological spectrum is diverse,and the bio-logical behavior is presented as a low to moderate malignancy,which provides the possibility for expanding the molecular classi-fication of CNS neuroepithelial tumor.

Objective:To investigate the efficacy of Mirabegron plus microecologics for the treatment of overactive bladder(OAB)in elderly women.Methods:In this prospective study, 104 patients diagnosed with OAB in the urology department of our hospital between February 2020 and December 2021 were recruited.The participants were randomly divided into two groups, with 52 in each.Group I was treated with Mirabegron alone(50 mg, qd)and Group Ⅱ was treated with Mirabegron(50 mg, qd)plus probiotics(3.5 g, bid)for 12 weeks.The efficacy was evaluated based on results before and after treatment, using measurements including the daily frequencies of urinary urgency, nocturia, daytime urination, urge incontinence and bladder capacity.The overactive bladder symptom score(OABSS), the quality-of-life score, and the number of lower urinary tract infections during treatment were also collected.Results at different treatment stages(week 4 and week 12)were compared within each group and between the two groups by Dunnett's test.Results:The symptoms of OAB in both groups were significantly improved after 12 weeks' treatment.In the group(Group Ⅱ)receiving the combination of two drugs, results from four measurements, the frequency of 24-hour urination(6.6 ± 0.7 vs.7.1 ± 1.2), the frequency of 24-hour urinary urgency(0.6 ± 0.6 vs.1.1 ± 0.7), the frequency of daily nocturia(0.8 ± 0.7 vs.1.3 ± 0.6)and the quality-of-life score(1.2 ± 0.7 vs.2.3 ± 0.8), were all significantly more favorable than in the group(Group I)treated with Mirabegron( P<0.05).The incidence of lower urinary tract infections and constipation was significantly reduced in the group with drug combination treatment. Conclusions:The efficacy of Mirabegron combined with microecologics for the treatment of OAB is better than that of Mirabegron alone, and the incidence of adverse events such as infections and constipation is also lower.

Objective:To evaluate the effect of penile dorsal extension and bandaging after concealed penis surgery.Methods:In this study, 80 children who underwent concealed penile correction were randomly divided into a dorsal extension bandaging group (experimental group) and a traditional bandaging group (control group) from Xuzhou Children’s Hospital Affiliated to Xuzhou Medical University during September 2016 to September 2019. The control group was applied with traditional sleeve-type bandaging, and the test group was treated with penile dorsal extension bandaging. A total of 75 children was summarized in this study. Among them, there were 38 children in the experimental group, with a mean age of 64 months. Thirty-seven children were in the control group, with a mean age of 70 months. The incidence of complications during hospitalization, the number of calls to medical staff within the first 24 hours after surgery, the pain score of the child during dressing removal, and the time taken for dressing removal were compared and recorded. Measurement data were analyzed by t-tests and enumeration data were analyzed by chi-square tests between groups. All data were analyzed using software SPSS 17.0. Results:The incidence of complications during hospitalization was 5.26%(2/38) and 10.81%(4/37) in the experimental and control groups, respectively. This difference was not statistically significant( χ2=0.784, P>0.05). In the dorsal extension bandaging group and the traditional bandaging group, the average number of calls to medical staff 24 hours after surgery was (0.87 ± 0.91) and (1.54 ± 1.02) times, the difference was statistically significant ( t=2.996, P=0.003); as for the highest pain scores of the children when the dressing was removed were 5.21 ± 1.19 and 7.24 ± 1.20, the difference was statistically significant ( t=5.697, P< 0.001); the time taken to remove the dressing was (3.21 ± 1.24) min, (7.56 ± 1.88) min, and the difference was statistically significant ( t=11.917, P<0.001). Conclusions:The penile dorsal extension bandaging method after concealed penis surgery can reduce the number of calls to medical staff, reduce the time of dressing removal and the degree of pain in children.

Objective:To reveal the mechanism of Mahuang Lianqiao Chixiaodou decoction and its disassembled formula for improving the skin barrier function in a mouse model of atopic dermatitis (AD).Methods:Sixty specific-pathogen free male BALB/c mice were randomly divided into the control group,model group,whole formula group (WF),exterior-releasing formula group (ERF),interior-clearing for-mula group (ICF),and positive control group (PC).A mouse model of AD was established using the semi-antigen 2,4-dinitrofluorobenzene induction method.The lesion scores,transepidermal water loss and pH,and skin histopathology of mice in each group were observed.The expressions of filaggrin,loricrin,and involucrin were detected by the streptavidin peroxidase immunohistochemical method and western blotting,and their mRNA expressions were detected by quantitative polymerase chain reaction.Results:Mice in the WF,ERF,ICF,and PC groups showed reduced skin lesion performance,improved histopathology,decreased skin lesion score,transepidermal water loss and pH,and upregulated ex-pressions of proteins including filaggrin,loricrin,and involucrin,and their mRNAs.The most obvious regulatory effect was observed in the WF group,followed by the ICF,ERF,and PC groups,accordingly.Conclusions:Mahuang Lianqiao Chixiaodou decoction and its disassembled formula can improve the skin barrier function in a mouse model of AD by upregulating filaggrin,loricrin,and involucrin,and their mRNA expressions,and the most optimal effect was noted in the WF group,followed by the ICF and ERF groups,which suggests that the effect of clearing heat and resolving dampness in improving the skin barrier function of AD is more obvious and is one of the key treatments for AD.

Objective:To evaluate the effect of penile dorsal extension and bandaging after concealed penis surgery.Methods:In this study, 80 children who underwent concealed penile correction were randomly divided into a dorsal extension bandaging group (experimental group) and a traditional bandaging group (control group) from Xuzhou Children’s Hospital Affiliated to Xuzhou Medical University during September 2016 to September 2019. The control group was applied with traditional sleeve-type bandaging, and the test group was treated with penile dorsal extension bandaging. A total of 75 children was summarized in this study. Among them, there were 38 children in the experimental group, with a mean age of 64 months. Thirty-seven children were in the control group, with a mean age of 70 months. The incidence of complications during hospitalization, the number of calls to medical staff within the first 24 hours after surgery, the pain score of the child during dressing removal, and the time taken for dressing removal were compared and recorded. Measurement data were analyzed by t-tests and enumeration data were analyzed by chi-square tests between groups. All data were analyzed using software SPSS 17.0. Results:The incidence of complications during hospitalization was 5.26%(2/38) and 10.81%(4/37) in the experimental and control groups, respectively. This difference was not statistically significant( χ2=0.784, P>0.05). In the dorsal extension bandaging group and the traditional bandaging group, the average number of calls to medical staff 24 hours after surgery was (0.87 ± 0.91) and (1.54 ± 1.02) times, the difference was statistically significant ( t=2.996, P=0.003); as for the highest pain scores of the children when the dressing was removed were 5.21 ± 1.19 and 7.24 ± 1.20, the difference was statistically significant ( t=5.697, P< 0.001); the time taken to remove the dressing was (3.21 ± 1.24) min, (7.56 ± 1.88) min, and the difference was statistically significant ( t=11.917, P<0.001). Conclusions:The penile dorsal extension bandaging method after concealed penis surgery can reduce the number of calls to medical staff, reduce the time of dressing removal and the degree of pain in children.