ObjectiveA middle cerebral artery occlusion (MCAO) mouse model is established by electrocoagulation of the middle cerebral artery. The study examines the mechanism by which exosomes (EXO) derived from human amniotic mesenchymal stem cells (hAMSCs) improve ischemic stroke and regulate neural ferroptosis-related injury. MethodsThirty-two SPF-grade male C57BL/6J mice aged 6 - 8 weeks were randomly divided into four groups (n=8 per group): sham group (Sham), model group (MCAO), MCAO plus normal saline group (MCAO+NaCl), and MCAO plus exosome group (MCAO+EXO). The mouse MCAO model was established by electrocoagulation of the middle cerebral artery. Mice in the Sham group underwent exposure of the middle cerebral artery without electrocoagulation. Twenty-four hours before MCAO induction, mice in the MCAO+EXO group received a tail vein injection of 100 μL of exosomes derived from the culture supernatant of hAMSCs at a concentration of 9.5×10¹¹ particles/mL. Mice in the MCAO+NaCl group were injected with an equal volume of normal saline via the tail vein. Twenty-four hours after model establishment, neurological deficits were evaluated using the Longa neurological deficit scoring system. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was performed to evaluate morphological changes of neurons in the ischemic brain regions. The contents of ferrous iron (Fe²⁺), malondialdehyde (MDA), total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) in the infarct core and peri-infarct regions were determined using microcolorimetric assays to evaluate differences among groups. The mRNA expression levels of ferroptosis-related factors, including nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the infarct core and peri-infarct regions were measured by real-time quantitative PCR. Protein expression levels of NRF2, SLC7A11, and GPX4 in the infarct and peri-infarct regions of each group were analyzed by Western blotting. ResultsCompared with the MCAO group, the Longa neurological deficit score was significantly reduced in the MCAO+EXO group (P<0.01). Prominent cerebral infarction was observed in the MCAO group, whereas the infarct volume ratio was markedly decreased in the MCAO+EXO group compared with the MCAO group (P<0.001). Histopathological analysis revealed that mice in the MCAO group exhibited obvious neuronal damage, including cytoplasmic vacuolar degeneration, nuclear pyknosis and fragmentation, unclear nuclear structure, and disorganized neuronal arrangement, compared with the Sham group. In contrast, neurons in the MCAO+EXO group showed relatively preserved morphology, with intact cellular structures and large, regular nuclei located centrally within the cells. Biochemical analysis demonstrated that Fe²⁺ and MDA levels in the infarct core and peri-infarct regions were significantly increased in the MCAO group compared with the Sham group (P<0.001). These levels were significantly reduced in the MCAO+EXO group compared with the MCAO group (P<0.01). In addition, total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) levels were markedly decreased in the MCAO group relative to the Sham group (P<0.01). Compared with the MCAO group, the MCAO+EXO group exhibited significantly increased levels of total GSH and GSH (P<0.001), while no significant change was observed in GSSG levels (P>0.05). Furthermore, both mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were significantly downregulated in the MCAO group compared with the Sham group (P<0.01, P<0.001). In contrast, both mRNA and protein expression levels of NRF2, SLC7A11, and GPX4 were significantly upregulated in the MCAO+EXO group compared with the MCAO group (P<0.05). ConclusionIn the mouse MCAO model, tail vein injection of exosomes derived from hAMSCs can improve motor function, reduce infarct area, protect neuronal cell morphology, and reduce the degree of nerve injury. Exosomes may exert a protective effect by activating the NRF2/SLC7A11/GPX4 pathway and reducing ferroptosis in neuronal cells of MCAO model mice.

OBJECTIVE To investigate the in vitro inhibitory mechanism of allicin against carbapenem-resistant Klebsiella pneumoniae(CRKP).METHODS The standard microbroth dilution method was employed to determine the minimum inhibitory concentration(MIC)of allicin against the experimental CRKP.Growth curve experiments were conducted to verify the relationship between allicin concentration and antibacterial action.RNA sequencing(RNA-seq)was performed on allicin-treated CRKP and a normal control group to explore the molecular mecha-nism of allicin inhibiting CRKP growth.RESULTS The MIC of allicin against CRKP was 140 mg/ml.A total of 1 775 differentially expressed genes were screened from the normal control group and the allicin-treated group,with 845 genes upregulated and 930 genes downregulated.The expression of bacterial phosphotransferase system(PTS)genes showed significant differences between the control group and the treated group(P＜0.05),and the expression levels of EIIA-EIID genes decreased(P＜0.05).CONCLUSION Based on the analysis of RNA-seq re-sults,it is found that allicin may inhibits the growth of CRKP by disrupting its sugar transport system,thereby affecting its biofilm formation.

Objective To explore the effective substances that specifically bind to RSV-F protein in the water extract of Forsythia suspensa,and verify its effect.Methods Surface plasmon resonance(SPR)affinity fishing technology was used to enrich,regenerate and recover the active components that specifically bind to RSV-F protein in the water extract of Forsythia suspensa.The recovered samples were analyzed and identified by UHPLC-MS/MS,and the active components that stably bind to F protein were screened by molecular docking,molecular dynamics simulation and SPR affinity detection,and then returned to animal experiments for pharmacodynamics verification.BALB/c mice were randomly divided into normal group,model group,positive control group,protocatechuic acid group and verbascoside group.In the experimental group,RSV was dripped into the nose to make models.Each drug group began to be given intragastric intervention on the day of modeling,and samples were taken after 4 days.HE staining was used to observe the pathological changes of lung tissue,and ELISA was used to detect the changes of inflammatory factors in lung tissue.Results A variety of active components in the aqueous extract of Forsythia suspensa could bind to RSV-F protein,among which the results of verbascoside and protocatechuic acid were more stable.In vivo pharmacodynamics experiments showed that the lung tissue of the model group had obvious pathological changes and the level of inflammatory factors increased,and the pathological state was improved after the intervention of verbascoside and protocatechuic acid compared with the normal group.Conclusion Forsythia suspensa can interfere with the expression of RSV-F protein through many active components such as verbascoside and protocatechuic acid,which may be the material basis of its anti-RSV.

Objective:To compare the efficacy and safety of crisaborole ointment 2% versus pimecrolimus cream 1% in the treatment of mild to moderate atopic dermatitis in children aged 2 years or older.Methods:A multicenter, randomized, open-label, controlled clinical trial was conducted. A total of 120 pediatric patients aged 2 - 17 years with mild to moderate atopic dermatitis were enrolled from departments of dermatology of 8 hospitals in China between March 2022 and February 2023. The participants were randomly assigned in a 1∶1 ratio to the crisaborole group and the pimecrolimus group, and received the treatment with crisaborole ointment 2% and pimecrolimus cream 1% respectively, twice a day for 4 weeks. Visits were scheduled at baseline/on day 1, as well as on days 8, 15, and 29. The primary efficacy outcome was the percentage of patients achieving the Investigator's Static Global Assessment (ISGA) success (defined as clear [0] or almost clear [1] on the ISGA scale, combined with ≥ 2‐grade improvement from baseline) on day 29. The secondary efficacy outcomes included changes in the Eczema Area and Severity Index (EASI) total scores from baseline to day 29, percentages of patients achieving ISGA improvement (defined as clear [0] or almost clear [1] on the ISGA scale), as well as changes in the Peak Pruritus Numerical Rating Scale (NRS) scores, Dermatology Life Quality Index (DLQI) /Infants' Dermatology Life Quality Index (IDLQI) /Children's Dermatology Life Quality Index (CDLQI) scores, and in the Dermatitis Family Impact (DFI) scores. Drug safety was evaluated according to the incidence of adverse events. Categorical data were compared using the chi-square test. Since measurement data did not follow a normal distribution, the rank sum test was used for comparisons of measurement data between groups.Results:A total of 106 children with mild to moderate atopic dermatitis were included in the per-protocol analysis set, with 52 in the crisaborole group (26 males and 26 females) and 54 in the pimecrolimus group (27 males and 27 females). There were no significant differences in age, disease duration, ISGA and EASI scores at baseline between the two groups (all P > 0.05). On day 29, 22 patients (42.31%) in the crisaborole group and 25 (46.30%) in the pimecrolimus group achieved ISGA success, with no significant difference between the two groups ( χ2 = 0.17, P = 0.68) ; 35 patients (67.31%) in the crisaborole group and 45 (83.33%) in the pimecrolimus group achieved ISGA improvement, also with no significant difference between the two groups ( χ2 = 3.68, P = 0.06) ; additionally, there were no significant differences in the EASI, pruritus NRS, DLQI/IDLQI/CDLQI, or DFI scores between the two groups (all P > 0.05). Adverse reactions to the two topical agents were mainly local reactions such as mild to moderate pain, itching, or worsening of itching, and no obvious systemic adverse reactions occurred. The incidence of drug-related adverse reactions was 46.15% (24 cases) in the crisaborole group and 37.04% (20 cases) in the pimecrolimus group, with no significant difference between the two groups ( χ2 = 0.91, P = 0.34) . Conclusion:The efficacy of crisaborole ointment 2% was comparable to that of pimecrolimus cream 1% in the treatment of mild to moderate atopic dermatitis in children aged ≥ 2 years, and it yielded early and rapid improvement in the quality of life of patients and their families, with good safety and tolerability profiles.

Objective:To investigate the incidence, clinical management and influencing factors of late-onset neutropenia (LON) in children with primary nephrotic syndrome (PNS) after Rituximab (RTX) treatment.Methods:A retrospective case-control study was conducted.The clinical data of PNS children who received RTX treatment at the First Affiliated Hospital of Xiamen University from March 2020 to August 2024 and were followed up for at least 6 months were collected.The RTX regimen was a single dose of 375 mg/m 2 with a maximum dose of 500 mg, and an additional dose was administered when the reexamination showed that peripheral blood CD19 + B cells were ≥ 1% or nephrotic syndrome relapsed.The patients were divided into an LON group and a non-LON group according to the presence or absence of LON, and then a comparison was made between the groups.The cumulative incidence of LON was estimated by the Kaplan-Meier method.Univariate and multivariate Logistic regression methods were used to identify the influencing factors of LON.Receiver operating characteristic (ROC) curves were plotted to assess the value of each influencing factor for predicting LON. Results:A total of 65 PNS patients were included.The incidence of LON was 19.3%(27/140) after 140 RTX treatment courses, and the first LON appeared 95.0 (64.0, 115.0) days after RTX treatment.Forty-nine patients received repeated RTX treatment.The incidence rates of LON after the first course, the second course, and the third course of RTX were 27.7%(18/65), 10.2%(5/49), and 18.8%(3/16), respectively, with no statistically significant difference ( χ2=5.764, P>0.05). There was also no significant difference in the incidence of LON between patients taking and not taking combined immunosuppressive agents after RTX treatment [35.3%(6/17) vs.37.8%(17/45), χ2=0.033, P>0.05]. Compared with the non-LON group, the LON group had a higher incidence of infections [48.1%(13/27) vs.15.0%(17/113), χ2=14.17, P<0.01], but no serious outcomes were observed in both groups.Multivariate Logistic analysis suggested that the age at treatment with RTX was an independent risk factor for LON after RTX treatment ( OR=0.763, 95% CI: 0.592-0.982). The area under the ROC curve of the age at treatment with RTX for predicting LON was 0.767 (95% CI: 0.628-0.906), with an optimal cutoff of 6.6 years, a sensitivity of 70.6%, and a specificity of 80.0%. Conclusions:The incidence of LON in PNS children after RTX treatment may be underestimated.Children who develop LON are at a higher risk of infections, but the prognosis is favorable.The age at treatment with RTX≤6.6 years is an independent risk factor for LON in PNS children.Close monitoring of neutrophil counts should be emphasized in younger PNS patients receiving RTX therapy.

Objective:To investigate the clinical efficacy and safety of a baby smoothing and special caring cream in reducing the recurrence of atopic dermatitis (AD) in infancy.Methods:A randomized controlled trial was conducted. Children with moderate AD (with overall investigator's global assessment [IGA] scores of 3 - < 4) were enrolled from Shunyi Maternal and Children′s Hospital of Beijing Children′s Hospital from April 2021 to June 2024. During the induction period, all children were topically treated with 0.1% hydrocortisone butyrate cream twice daily on the lesional skin, as well as with a baby smoothing and special caring cream at least twice daily throughout the body; at the 2-week visit, patients with an IGA score of ≤ 1 point entered the maintenance phase, while those with an IGA score of > 1 point continued the treatment for another 2 weeks; at the 4-week visit, patients with an IGA score of ≤ 1 point entered the maintenance phase, while those still with an IGA score of > 1 point were withdrawn from the study, and received conventional treatment. Patients who entered the maintenance period were randomly divided into the test group and the control group in a 1∶1 ratio using a random number table. In the test group, the hydrocortisone butyrate cream was discontinued, while the baby smoothing and special caring cream was continued twice daily for 8 consecutive weeks; in the control group, both the hydrocortisone butyrate cream and the baby smoothing and special caring cream were discontinued. IGA and Scoring AD (SCORAD) scores were assessed by clinicians at weeks 4 and 8 in the maintenance phase, while the patient-oriented eczema measure (POEM) score was evaluated weekly by patients' parents. The Kaplan-Meier survival analysis and Breslow test were used to compare recurrence rates in the two groups (the primary efficacy outcome), and a generalized estimating equation model was used to evaluate the changes in IGA, SCORAD, and POEM scores in the two groups (the secondary efficacy outcomes). Adverse reactions were monitored throughout the study to evaluate safety.Results:A total of 68 children with moderate AD aged from 3 months to 2 years were included. There were 38 females and 30 males, aged 11.72 ± 6.03 months. Fifty-two patients entered the maintenance phase; 2 were lost to follow-up, and 50 were included in the per-protocol set, with 28 in the test group and 22 in the control group. The recurrence rate during the maintenance phase was 7.14% (2/28) in the test group and 31.82% (7/22) in the control group, showing a significant difference between the two groups ( χ 2 = 5.08, P = 0.032). At weeks 4 and 8 in the maintenance phase, the IGA scores were significantly lower in the test group than in the control group (Wald χ 2 = 5.06, P = 0.024), whereas the SCORAD scores showed no significant differences between the two groups (Wald χ 2 = 2.92, P = 0.087). During weeks 1 - 8 in the maintenance phase, the POEM scores showed no significant differences between the two groups or over time (both P > 0.05), while the two groups showed different change trends in POEM scores over time (Wald χ 2interaction = 55.37, Pinteraction < 0.001). Throughout the entire study period, no adverse reactions were observed among all 68 subjects. Conclusion:With a high safety profile, the baby smoothing and special caring cream could reduce the recurrence rate during the maintenance phase, showing promise as an adjuvant therapy for the maintenance treatment of AD in infancy, and is worthy of clinical application.

OBJECTIVE To investigate the in vitro inhibitory mechanism of allicin against carbapenem-resistant Klebsiella pneumoniae(CRKP).METHODS The standard microbroth dilution method was employed to determine the minimum inhibitory concentration(MIC)of allicin against the experimental CRKP.Growth curve experiments were conducted to verify the relationship between allicin concentration and antibacterial action.RNA sequencing(RNA-seq)was performed on allicin-treated CRKP and a normal control group to explore the molecular mecha-nism of allicin inhibiting CRKP growth.RESULTS The MIC of allicin against CRKP was 140 mg/ml.A total of 1 775 differentially expressed genes were screened from the normal control group and the allicin-treated group,with 845 genes upregulated and 930 genes downregulated.The expression of bacterial phosphotransferase system(PTS)genes showed significant differences between the control group and the treated group(P＜0.05),and the expression levels of EIIA-EIID genes decreased(P＜0.05).CONCLUSION Based on the analysis of RNA-seq re-sults,it is found that allicin may inhibits the growth of CRKP by disrupting its sugar transport system,thereby affecting its biofilm formation.

Objective:To investigate the operation opportunity for vascular rings in infants and assess the impact of prenatal and postnatal integrated management strategies on treatment outcomes.Methods:A retrospective analysis was conducted on the clinical data of 75 infants with vascular rings who underwent surgical treatment at the Third Affiliated Hospital of Zhengzhou University from January 2016 to December 2023. Among them, 54 were males and 21 were females, with a median age at surgery of 1.7 (0.7-6.9) months and a median weight of 5.3 (3.5-8.0) kg. Vascular rings malformation was diagnosed by real-time three-dimensional color doppler echocardiography during pregnancy in 51 cases. Preoperatively, 28 cases presented with respiratory or digestive system-related symptoms, and 26 cases had a history of hospitalization due to related symptoms. All patients underwent preoperative cardiac CTA+ CTVE and three-dimensional reconstruction examinations, and 56 cases showed varying degrees of airway compression and stenosis on imaging. Among them, 10 patients presented with preoperative stridor and respiratory distress; fiberoptic bronchoscopy performed after anesthesia induction confirmed significant tracheal compression/stenosis. One patient was ventilator-dependent preoperatively, and bronchoscopy revealed main bronchomalacia. The cohort included: Complete vascular rings (62 cases of double aortic arch, 10 cases of right aortic arch with aberrant left subclavian artery and left-sided ductus arteriosus/ligamentum) and incomplete vascular rings (3 cases of pulmonary artery sling). Additionally, 5 cases had associated Kommerell’s diverticulum, and 12 had intracardiac malformation. All patients successfully completed surgery, and those with intracardiac malformation underwent extracorporeal circulation and primary radical surgery concurrently. Based on prenatal diagnosis and implementation of prenatal and postnatal integrated management, patients were divided into an observation group (prenatal and postnatal integrated management group) and a control group (postnatally diagnosed group). The perioperative data of the two groups were compared to analyze the surgical outcomes and the advantages of prenatal and postnatal integrated treatment.Results:All 75 patients successfully completed surgery. Preoperatively, 56 cases (74.66%) presented with varying degrees of tracheal stenosis. Except for 1 case that died after abandoning treatment and 1 case that underwent tracheal surgery due to repeated failed ventilator weaning, all other patients were successfully discharged from the hospital. The overall mortality rate was 1.33%, and the rate of tracheal surgery was 1.35%. The age and weight at surgery in the observation group were lower than those in the control group ( P<0.05), and the proportion of preoperative hospitalization history was lower in the observation group ( P<0.05). No significant differences were observed between the two groups in terms of tracheal compression and stenosis, postoperative monitoring time, operation time, ventilator time, and risk of postoperative complications ( P>0.05). Conclusion:Tracheal stenosis is a common complication in children with vascular rings. Early surgical intervention is recommended for complete vascular rings and pulmonary artery slings (as an incomplete ring). Timely prenatal diagnosis of vascular ring anomalies combined with the implementation of an integrated prenatal-postnatal management strategy can significantly reduce the risk of preoperative hospitalization due to related symptoms and may lower the risk of subsequent tracheal surgery, potentially improving long-term prognosis.

Objective:To investigate the treatment regimens, therapeutic effects, and adverse events of immune checkpoint inhibitors (ICI) in the treatment of progressive multifocal leukoencephalopathy (PML).Methods:A retrospective analysis was performed on patients with PML who received ICI treatment at the Department of Neurology of Peking Union Medical College Hospital from October 2020 to September 2024. The patients′ demographic characteristics, baseline immune status, clinical manifestations, laboratory tests, treatment regimens, and clinical outcomes were analyzed.Results:A total of 2 male patients with PML, aged 40 and 59 years, received ICI treatment. One patient had underlying combined immunodeficiency, while the other had acute myeloid leukemia subtype M2 and had previously undergone umbilical cord blood stem cell transplantation. Both patients were treated with pembrolizumab (dosage range: 2-3 mg/kg, administered every 3-4 weeks), receiving a total of 2-4 courses of treatment. In terms of therapeutic effects, both patients showed significant improvement. Regarding adverse events, 1 patient experienced immune-related adverse event (irAE) of immune-related pneumonia combined with immune-related hypophysitis.Conclusions:ICI may be an effective treatment option for PML. However, the use of ICI may be accompanied by the occurrence of irAE, necessitating close monitoring during treatment.

Objective:To explore the relationship between the dynamic alterations of immune cell subsets in a patient with autoimmune encephalitis and the clinical relapses.Methods:For a patient with multiple relapses of anti-N-methyl-D-aspartate receptor encephalitis superimposed with myelin oligodendrocyte glycoprotein antibody-associated disease who visited Peking Union Medical College Hospital on July 18, 2018, the lymphocyte subsets were monitored dynamically over a long period and the relationship between lymphocyte subsets and the relapses was summarized.Results:The 38-year old male patient experienced a total of 5 episodes (including 4 relapses) during the six-year immunotherapy and follow-up process, and responded well to first-line and maintenance immunotherapy. His relapses occurred during drug reduction. Finally, the neurological symptoms resolved after rituximab treatment. A total of 39 tests of peripheral blood lymphocyte subsets were conducted during the follow-up, and 5 peaks of elevated CD19 positive B cells, 9 peaks of elevated CD3 positive T cells and 11 peaks of elevated natural killer cells were observed. The peak period of peripheral blood CD19 positive B cells exactly coincided with clinical relapses. While the consistency rates of clinical relapse and the peaks of peripheral blood CD3 positive T cells and natural killer cells were 5/9 and 2/11, respectively.Conclusions:The dynamic alterations of CD19 positive B cells in peripheral blood are correlated with the clinical relapse of autoimmune encephalitis and can predict clinical relapse.