Objective A rat model of cerebral small vessel disease(CSVD)was established by unilateral injection of a single dose of sodium laurate into the internal carotid artery.The effectiveness of the model was assessed by behavior scoring and analysis of serum-related indicators,cerebral infarction volume,cerebral microvascular density,hemodynamics,brain histopathology and the expression of blood-brain barrier(BBB)-related proteins.Methods SPF-grade male SD rats were divided randomly into a control group and a model group(n=6 per group).The model group received a single injection of 100 μL of sodium laurate(2 g/L)via the internal carotid artery,while the control group underwent the same surgical procedure but received an equal volume of saline.Neurobehavioral assessments were conducted using the Longa score and postural reflex test.Serum homocysteine(HCY)levels were measured by enzyme-linked immunosorbent assay.Cerebral infarction volume was detected by magnetic resonance imaging and changes in cerebral vascular density were observed by cerebrovascular imaging.The resistance index(RI)and perfusion index(PI)were measured by ultrasonography.Histopathological changes in brain tissue were evaluated by hematoxylin and eosin(HE)staining.Expression of the cerebral microvascular marker CD31 and tight junction proteins ZO-1 and Occludin in brain cortex tissue were detected by immunohistochemical staining.Results The Longa score,postural reflex score(P＜0.05),and cerebral infarction volume were significantly increased(P＜0.05)while the cerebral vascular density was decreased in the model group compared with the control group.Serum HCY levels,carotid RI,and PI values were all significantly increased in the model group(P＜0.05).HE staining revealed solidified neuronal nuclei and enlarged perivascular spaces in the brain cortex in the model group.Immunohistochemical staining revealed that CD31,ZO-1,and Occludin expression were significantly reduced in the brain cortex in the model group compared with the control group(P＜0.05).Conclusions A rat model of CSVD can be established rapidly and effectively by a single unilateral injection of high-concentration sodium laurate via the internal carotid artery.This model is characterized by neurobehavioral abnormalities,cerebral infarction,insufficient cerebral blood supply,reduced vascular density,and disruption of the BBB,suggesting that it may serve as an effective rat model for the study of CSVD.

Objective:To analyze the risk factors for human cytomegalovirus (HCMV) infection in pediatric recipients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Clinical data of children who underwent first allo-HSCT were retrospectively analyzed from March 2017 to November 2024. A total of 259 pediatric allo-HSCT recipients were analyzed through comparing HCMV infection group (n=115) and Non-HCMV infection group (n=144). Clinical characteristics were compared, and risk factors for HCMV infection were analyzed using univariate and multivariate logistic regression.Results:The result of univariate analysis showed that adrenoleukodystrophy (ALD), length of hospitalization, duration of antiviral therapy, and bacterial infection were significantly associated with HCMV infection in pediatric allo-HSCT recipients ( P<0.05). The result of multivariate analysis showed that ALD was an independent protective factor against HCMV infection of allo-HSCT recipients ( P<0.05) [OR=0.22, 95% CI: 0.06-0.86], while umbilical cord blood transplantation (UCBT) was an independent risk factor for HCMV infection in allo-HSCT recipients ( P<0.05) [OR=6.13, 95% CI: 1.34-28.04]. When the area under the ROC curve (AUC) for predicting post-transplant relapse based on HCMV viral load was 0.75 (95% CI: 0.55-0.94, P=0.014) and at the cutoff value of 3×10 3 copies/ml, the sensitivity and specificity for predicting relapse were 81.13% and 66.67%, respectively. Conclusions:HCMV infection in pediatric allo-HSCT recipients may lead to longer hospitalization and increased risk of relapse.

Objective:To determine the epidemiological characteristics of human bocavirus (HBoV) in children with acute respiratory infections (ARI) in Bengbu City, Anhui Province, in 2024.Methods:Nasopharyngeal swab samples were collected from 269 children with ARI in Bengbu City, Anhui Province, in 2024. Seventeen respiratory pathogens were screened using quantitative fluorescence PCR. For HBoV-positive samples, the VP1/VP2 structural gene fragments of HBoV were amplified and sequenced for genetic evolutionary analysis.Results:Among the 269 nasopharyngeal swab samples from children with ARI, the overall detection rate of respiratory pathogens was 48.33% (103/269). The top three pathogens with the highest detection rates were: Influenza A virus (FluA): 10.04% (27/269), Respiratory syncytial virus (RSV): 8.18% (22/269), Human bocavirus (HBoV): 7.43% (20/269). The age distribution of HBoV-infected children showed that the detection rate was highest in the 0-2 years age group (50%, 10/20), followed by the 3-5 years age group (25%, 5/20) and the over 6 years age group (25%, 5/20). However, there was no statistically significant difference in viral detection rates among the age groups. Genetic evolutionary analysis based on VP1/VP2 revealed that all 13 HBoV strains were of the HBoV-1 genotype.Conclusions:HBoV is one of the major pathogens causing ARI in children in Bengbu City, Anhui Province, in 2024, with HBoV-1 being the predominant genotype. Additionally, infants aged 0-2 years are the most susceptible population to HBoV infection.

Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.

Objective:To evaluate the efficacy of corticosteroids in male children with Duchenne muscular dystrophy (DMD), and provide evidence for the rational clinical use of medication.Methods:This was a multicenter medical record series study which conducted from January 15 th to March 14 th, 2025. A total of 53 male children with DMD admitted to the Department of Rehabilitation of Children′s Hospital, Zhejiang University School of Medicine, Children′s Hospital of Chongqing Medical University and Affiliated Children′s Hospital of Soochow University from 2020 to 2024 were enrolled. Clinical data, corticosteroid usage, and the follow-up data were collected. The North star ambulatory assessment (NSAA) was used as the primary efficacy indicator. Generalized estimating equations (GEE) exchangeable working matrices were used for longitudinal analysis, and the least squares mean were used to compare the change trend of the efficacy evaluation index across different medication durations. Results:The age at the initiation of corticosteroid treatment was (6.3±1.9) years. The follow-up duration was 1.2 (0.9, 2.2) years. After treatment, the raw scores and linear scores of NSAA were both significantly higher than those before treatment ((22±7) vs. (19±5) points, (60±16) vs. (53±8) points; t=3.98, 3.69; both P<0.001). The 10 meter running time and time rising from floor were both shorter than those before treatment (6 (4, 8) vs. 7 (6, 9) s, 5 (3, 6) vs. 6 (5, 9) s; Z=2.62, 3.47; both P<0.01). GEE model analysis revealed all nonlinear correlation between motor function (NSAA linear score, 10-meter running velocity, and rising from floor velocity) and the duration of corticosteroid treatment (all P<0.05). Least squares mean comparison all showed that the medication effect first increased and then decreased with duration, reaching the peak at 1.1-2.0 years after treatment (all P<0.05). Conclusions:Corticosteroids can improve the motor function in male children with DMD, with the maximum treatment effect occurring 1 to 2 years after the initiation of treatment. It is necessary to comprehensively leverage time-varying efficacy of corticosteroids to optimize individualized treatment regimens for maximal motor function benefits in children with DMD.

Objective To investigate the effects of integrating pelvic floor magnetic stimulation and medium-frequency pulsed electrotherapy with biofeedback on alleviating obstructed defecation symptoms in patients diagnosed with pelvic floor spasm syndrome.Methods A total of 133 patients diagnosed with pelvic floor spasm syndrome at the Gastrointestinal Surgery Outpatient and Inpatient Departments of the Third Hospital of Hebei Medical University between January 2017 and January 2025 were enrolled in this study.All patients underwent two sessions of conventional biofeedback therapy.According to the additional treatments they received,the patients were catego-rized into four groups:the Control group(received biofeedback therapy only);the Magnetic Stimulation group(received two additional sessions of pelvic floor magnetic stimulation);the Electrical Stimulation group(received two additional sessions of medium-frequency pulse electrical therapy);and the Magnetic-Electric Combined group(received two additional sessions of both pelvic floor magnetic stimulation and medium-frequency pulse electrical therapy).The improvement in obstructed defecation symptoms was evaluated before treatment,after treatment,and at the 3-month follow-up across all four groups.Results A total of 133 patients diagnosed with pelvic floor spasm syndrome were enrolled in this study.After a 3-month follow-up period,8 patients were lost to follow-up(6.0%).Consequently,125 patients completed the follow-up and were included in the final analysis.These patients were distributed across four groups:the control group(n=32),the magnetic stimulation group(n=30),the electrical stimulation group(n=31),and the combined magneto-electric stimulation group(n=32).Compared with biofeedback therapy alone,combination therapy led to a greater reduction in the need for manual assistance during defecation,the sensation of incomplete evacuation,and the requirement for straining during defecation,with the most pronounced improvements observed in the combined magneto-electric stimulation group.Conclusions The integration of biofeedback,pelvic floor magnetic stimulation,and medium-frequency pulse electrotherapy can significantly alleviate symptoms including straining during defecation,the sensation of incomplete bowel evacua-tion,and reduce the need for manual assistance.Moreover,this combined approach contributes to the stabilization and maintenance of therapeutic effects in the short term.

Objective To explore the associations of dietary patterns with bone mineral density(BMD)and osteoporosis(OP)in patients with knee osteoarthritis(KOA)in south China,identify the dietary patterns that could reduce the risk of OP,and provide a reference for prevention of OP.Methods Data from 600 KOA patients who were enrolled in the Pearl River Osteoarthritis Cohort(PROC)between April 2020 and April 2024 were extracted,including general characteristics,dietary questionnaire and BMD test.Factor analysis was employed to identify dietary patterns.Scores of dietary pattern were treated as continuous or categorical variables(quartile-based groups).Multivariable linear regression was used to assess associations between scores of dietary pattern and BMD,while the relationships with OP were analysed by binary logistic regression.Results The prevalence of OP in KOA patients was 19.8%.Factor analysis extracted eight dietary patterns:seafood-nuts,livestock-poultry meats,coarse grains-fruits,wheat-eggs,fish-dairy-vegetables,legumes-fruits,rice-wheat flour,and soy-dairy.After adjusting for confounders,it was found that the pattern of coarse grains-fruits was positively correlated with the BMD of left femoral neck and lumbar spine(P<0.01)and the fish-dairy-vegetables pattern was positively correlated with the BMD of lumbar spine(P<0.01).When pattern scores were discretized into quartiles,the legumes-fruits pattern(Q3 vs.Q1)was found in association with a 68.2%reduction in the risk of OP(P<0.05).Conclusion This study has identified that the pattern of legumes-fruits may lower a risk of OP,while the patterns of coarse grains-fruits and fish-dairy-vegetables correlate with higher BMD.The diet patterns are potentially beneficial to prevention of OP.For bone health,a balanced diet rich in legumes,fruits,vegetables,coarse grains,fish,nuts and dairy is recommended.

BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide （LPS）-induced inflammation of human colorectal adenocarcinoma （Caco-2） cells by inhibiting apoptosis via activating the tumor necrosis factor （TNF） receptor superfamily member 6 （Fas）/Fas ligand （FasL） pathway. MethodsCaco-2 cells were allocated into blank， model （LPS， 10 mg·L^-1）， Shaoyaotang-containing serum （5%， 10%， 15%， 20%）， and Fas inhibitor （KR-33493， 20 mmol·L^-1） groups. Except the blank group， the other groups were stimulated with 10 mg·L^-1 LPS for 24 h for the modeling of inflammation. After successful modeling， the blank， Fas inhibitor， and model groups were treated with blank serum， and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 （CCK-8） method. The levels of interleukin （IL）-6， IL-1β， and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas， FasL， cysteinyl aspartate-specific proteinase （Caspase）-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax） were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultsCompared with the blank group， the model group presented a decrease in cell survival rate （P<0.01）. Compared with that in the model group， the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%， 15%， and 20% Shaoyaotang-containing serum groups （P<0.01）. Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group， 10%， 15%， and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group， the model group showed risen levels of IL-6， IL-1β， and TNF-α （P<0.01）， an increased apoptosis rate （P<0.01）， up-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.01）， and down-regulated protein and mRNA levels of Bcl-2 （P<0.01）. Compared with the model group， the Fas inhibitor group and the 10%， 15%， and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6， IL-1β， and TNF-α （P<0.01）， decreased apoptosis rates （P<0.01）， down-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and up-regulated protein and mRNA levels of Bcl-2 （P<0.05， P<0.01）. In addition， the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6， IL-1β， and TNF-α （P<0.05， P<0.01）， lower apoptosis rates （P<0.05， P<0.01）， lower protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and higher protein and mRNA levels of Bcl-2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells， down-regulate the protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax， and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.

ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang （SYD） on the lipopolysaccharide （LPS）-induced damage of tight junction proteins in the human colorectal adenocarcinoma （Caco-2） cell model of inflammation via the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） pathway. MethodsCaco-2 cells were grouped as follows： Blank， model （LPS， 10 mg·L^-1）， SYD-containing serum （10%， 15%， and 20%）， and inhibitor （Fasudil， 25 μmol·L^-1）. After 24 hours of intervention， the cell viability in each group was examined by the cell-counting kit 8 （CCK-8） method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 （ET-1）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of RhoA， ROCK2， claudin-5， and zonula occludens-1 （ZO-1） in cells of each group. ResultsCompared with the blank group， the model group showcased a marked reduction in the cell viability （P<0.01）， elevations in the levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， declines in both mRNA and protein levels of ZO-1 and claudin-5 （P<0.01）， and rises in mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Compared with the model group， the Shaoyaotang-containing serum （10%， 15%， and 20%） groups had enhanced cell viability （P<0.01）， lowered levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， up-regulated mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and down-regulated mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Moreover， the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， higher mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and lower mRNA and protein levels of RhoA and ROCK2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway.