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.

ObjectiveTo compare female C57BL/6J and BALB/c mice models of ulcerative colitis (UC) induced by dextran sulfate sodium salt (DSS), providing a reference for selecting female animal models in studies of innovative drugs for UC treatment. MethodsA total of 48 female mice aged 6–8 weeks, including 24 C57BL/6J and 24 BALB/c mice, were divided into four groups: control group, UC-3d group (2.5% DSS drinking for 3 days), UC-7d group (2.5% DSS drinking for 7 days), and UC-withdrawal group (2.5% DSS drinking for 7 days followed by 3 days of pure water replacement), with 6 mice in each group. Body weight, fecal occult blood, and stool characteristics were recorded daily, and disease activity index (DAI) scores were calculated. At the end of DSS induction, mice were gavaged with fluorescein isothiocyanate (FITC)-labeled dextran solution before tissue collection. Four hours later, under deep anesthesia induced by inhalation of 4% isoflurane, blood samples were collected from the retro-orbital venous plexus, and euthanasia was performed by cervical dislocation. The colon was then separated immediately, and its length from the anus to the ileocecal junction was measured. Colon tissues were fixed, embedded, and sectioned, and they were evaluated using hematoxylin-eosin (HE) staining and alcian blue-periodic acid-Schiff (AB-PAS) staining to comprehensively assess the degree of colonic inflammation. The spleen was also collected and weighed, and the spleen coefficient was calculated. Serum FITC fluorescence intensity and D-lactic acid concentration were measured to evaluate intestinal permeability and barrier injury. The grooming duration, the number of buried marbles, the time spent in the center of the open field, and the total distance traveled were measured to evaluate anxiety/depression-like behaviors in mice. ResultsFecal occult blood appeared in both C57BL/6J and BALB/c mice after 3 days of ad libitum access to DSS. Compared with the control group, there was no significant change in the weight loss rate of female BALB/c mice in the UC-7d and UC-withdrawal groups (P > 0.05), but the DAI score increased (P < 0.01). Conversely, both the weight loss rate and DAI score increased for female C57BL/6J mice in the UC-7d and UC-withdrawal groups (P < 0.01). In female C57BL/6J mice, both the UC-7d and UC-withdrawal groups had higher spleen coefficients than the control group (P < 0.05). Additionally, in these groups, colon length was significantly reduced (P < 0.01), FITC fluorescence intensity in serum was markedly increased (P < 0.01), obvious inflammatory reaction areas appeared in the colon, and the goblet cells and acidic mucus layer were damaged. In contrast, in female BALB/c mice, no significant differences were observed in colon length, serum FITC fluorescence intensity, or D-lactic acid concentration between the UC-7d and UC-withdrawal groups compared with the control group (P > 0.05). Only a few inflammatory cells and damaged goblet cells were observed in the colonic mucosal layer, and no other obvious pathological changes were detected. In addition, female C57BL/6J mice in the UC-7d group showed a significantly reduced total distance in the open field and fewer buried marbles (P < 0.05), indicating depression-like behavior, as well as prolonged grooming duration (P < 0.05), indicating anxiety-like behavior. ConclusionFemale BALB/c mice are not sensitive to 2.5% DSS and do not show typical colonic pathological changes of UC. In contrast, ad libitum access to 2.5% DSS for 7 days successfully induces typical UC pathological symptoms and colonic pathological changes in female C57BL/6J mice. These changes persist for 3 days after DSS withdrawal, and the mice also exhibit symptoms of anxiety/depression. The results suggest that this model can be used for research on the pathogenesis and therapeutic drugs of UC.

Objective To screen hub genes and signaling pathways associated with acute ischemic stroke(AIS)using bioinformatics methods,identify potential biomarkers,and provide new evidence for the mechanism research of AIS.Methods The gene expression dataset GSE37587 of AIS patients and healthy controls was obtained from the public database gene expression omnibus(GEO).The differentially expressed genes(DEGs,|log2 FC|≥1.2,FDR＜0.05)were screened using the limma package.The enrichment analysis of GO/KEGG was performed with the DAVID database.The weighted correlation network analysis(WGCNA)was used to construct a gene co-expression network for screening key modules.Then,a protein-protein interaction(PPI)network was constructed based on the STRING database and Cytoscape software to identify hub genes.The dataset GSE16561 was used to validate.Meanwhile,the clinical samples from 30 AIS patients and 30 healthy controls visited Zibo First Hospital from January to May 2025 were validated by the real time fluorescence quantitative PCR(qRT-PCR).The diagnostic efficacy was evaluated using the receiver operating characteristics(ROC)curve.Results A total of 653 DEGs were identified,including 252 up-regulated and 401 down-regulated genes.They were mainly enriched in biological processes such as ribosome biogenesis,endoplasmic reticulum protein processing,and oxidative phospho-rylation,as well as signaling pathways such as viral infection-related pathways and PD-L1/PD-1 checkpoint pathways in cancer.The core genes in the light green module identified by the WGCNA analysis were significantly enriched in the pathways such as mitophagy,ribosome,and endocytosis.The hub genes such as RPL34 and DDIT3 were screened from the PPI network,and their expression levels were significantly correlated with AIS.The analysis of the ROC curve showed that the areas under the ROC curve(AUCROC)of the hub genes for the diagnosis of AIS were 0.78-0.82,which had high clinical application value.Conclusion Ribosomal proteins,endoplas-mic reticulum stress-related genes,and viral infection response pathways are key molecular events in the occurrence of AIS.The genes such as RPL34 and DDIT3 are expected to be potential biomarkers for AIS,providing experimental evidence for the development of di-agnostic markers.

Objective To establish an efficient,sensitive,and accurate ion chromatography method for the simultaneous determination of three impurity ions—bromide(Br-),nitrite(NO2-),and oxalate(C2 O42-)—in oral rehydration salts powder(Ⅲ).These impurity ions may pose potential toxic side effects on children's health;therefore,ensuring their content within safe limits is crucial for safeguarding pediatric medication safety.Methods A Metrosep A Supp 5 column(250 mm×4.0 mm,5 μm),packed with quaternary ammonium-bonded polyvinyl alcohol was used.The eluent consisted of carbonate buffer:Mobile phase A was a mixture of 3.2 mmol·L-1 sodium carbonate and 1.0 mmol·L-1 sodium bicarbonate(50∶50,V∶V),and mobile phase B was a mixture of 32 mmol·L-1 sodium carbonate and 10 mmol·L-1 sodium bicarbonate(50∶50,V∶V),with gradient elution.The column temperature was set at 35℃,and the flow rate was 0.7 mL·min-1.Limits for the three impurity ions were set according to the International Council for Harmonisation guidelines(ICH M7(R1)and ICH Q3B),and the method was validated by testing six actual samples.Results Within the concentration range of 0.02-0.50 μg·mL-1,the linear correlation coefficients(R2)for the three impurity ions were all greater than 0.999.The detection limits ranged from 1.2 to 3.4 ng·mL-1,and the spiked recovery rates were between 92.0%and 102.0%.Results from the six actual samples demonstrated that the method effectively controlled the content of impurity ions.Conclusion This study successfully established a gradient elution ion chromatography method for the simultaneous determination of three impurity ions in Oral rehydration salts powder(Ⅲ).The method is highly sensitive,specific,and accurate.

Objective To investigate the status of feeding intolerance in patients with enteral nutrition after lung transplantation and analyze its influencing factors,to provide a reference for formulating a reasonable enteral nutrition plan and improving patients'nutritional status.Methods Convenient sampling method was used to retrospectively collect the clinical data of 115 patients who received enteral nutrition support after lung transplantation and were hospitalized in the ICU of a tertiary hospital in Guangdong Province from August 2022 to November 2023.According to the occurrence of feeding intolerance during ICU hospitalization,the patients were divided into a feeding tolerance group and a feeding intolerance group.Univariate and logistic regression analysis were used to analyze the influencing factors of feeding intolerance patients with enteral nutrition after lung transplantation.Results Within 7 days of initiating enteral nutrition,a total of 63 patients developed feeding intolerance,with an incidence of 54.78％.Among them,the incidence of feeding intolerance was relatively high within 1 to 3 days after initiating enteral feeding.The clinical manifestations of feeding intolerance were diarrhea,bloating,gastric retention,vomiting/regurgitation,among which the diarrhea was the highest incidence(87.30％).Logi-stic regression analysis showed that intraoperative net balance volume(OR=0.999),intraoperative blood transfusion(OR=1.001)volume and diabetes history(OR=0.170)were independent influencing factors for feeding intolerance in patients with enteral nutrition after lung transplantation(P＜0.05).Conclusion There was a high incidence of feed-ing intolerance in patients with enteral nutrition after lung transplantation.Patients undergoing lung transplantation who have a high net intraoperative fluid balance,receive a low volume of intraoperative blood transfusions,and have a history of diabetes are at a lower risk of developing feeding intolerance when receiving postoperative enteral nutrition.When starting enteral nutrition,medical staff should dynamically evaluate the risk factors of feeding intolerance,screen high-risk patients as early as possible,and formulate reasonable enteral nutrition programs to improve the nutritional status of patients and promote their rehabilitation.

In order to explore the relationship between ubiquitin proteasome system(UPS)and au-tophagy after macrophage was infected with Brucella.On the one hand,the levels of LC3 Ⅱ、P62 and 20S proteasomes in cell supernatant and peritoneal fluid were determined respectively after RAW264.7 cells were infected and BALB/c mice were intraperitoneal inoculated with Brucella suis(B.suis).The results displayed that UPS was activated firstly,followed by autophagy after B.suis infection.On the other hand,we prepared the UPS-inhibited-cells and ALP-inhibited-cells by lacta-cystin and 3-methy ladenine(3-MA),and then the cells were infected by B.suis and the levels of LC3 Ⅱ、P62 与 20S proteasomes in cell supernatant were determined.The results showed that the ALP function was significantly improved when the effect of M

Bispecific antibodies(BsAbs),as an important recent innovation in the field of tumor immunotherapy in recent years,can simultaneously or sequentially target different antigens or two different epitopes of the same antigen.Compared with traditional monoclonal antibodies,they can produce superior therapeutic effects.This article reviews the progress in clinical applications and safety research of BsAbs in cancer therapy,revealing that they(such as blinatumomab,glofitamab,teclistamab,amivantamab,etc.)exhibit significant therapeutic efficacy against hematological malignancies,lung cancer,cervical cancer,melanoma,and other cancers.For cytokine release syndrome(CRS)induced by BsAbs,prophylactic or pre-emptive medication is commonly administered in clinical practice;for neurotoxicity and infections triggered by BsAbs,clinical practice necessitates rigorous monitoring of patients'vital signs and the provision of essential treatments.In addition,different BsAbs exhibit variations in escalation dose,infusion rate,storage duration,and equipment requirements.Therefore,strict adherence to the instructions in the drug package inserts is essential during clinical operations to ensure safety and therapeutic efficacy.In the future,more multicenter trials need to be conducted to validate the efficacy and safety of BsAbs across different tumor types and patient populations,and long-term follow-up data should be accumulated to optimize treatment cycles and dosage regimens.

Objective:To evaluate the functional outcomes and postoperative complications associated with modified Toupet-like (mToupet-like) anastomosis following proximal gastrectomy for patients with gastric tumors.Methods:After proximal gastrectomy, barbed sutures (2-3 stitches) in the seromuscular layer were used to secure the anterior wall of the stomach at a distance of 1-2 cm from the closure line and the posterior wall of the esophagus at a distance of 5.0 cm from the closure line. The remnant stomach was then positioned posterior to the esophagus on the greater curvature side. Esophagogastric anterior wall anastomosis (manual or circular stapling) was performed at the greater curvature of the remnant stomach, 3 cm distal to the gastroesophageal fixation point. A Toupet-like folding procedure was conducted by folding the reconstructed gastric fundus and wall anteriorly from behind the esophagus and embedding the esophagus within a 270° wrap at the site of stomach-esophagus fixation.Results:Twelve patients with gastric tumors underwent proximal partial gastrectomy with mToupet-like anastomosis in the Department of Gastric Surgery at Zhejiang Cancer Hospital from January to March 2024. Among them, 10 diagnosed as upper gastric adenocarcinoma, and 2 diagnosed as gastric gastrointestinal stromal tumors. The cohort included nine male patients and three female patients, aged 46 to 77 years old, with a body mass index (BMI) ranging from 19.7 to 27.3 kg/m2. The maximum tumor diameter was less than 4 cm, and the predicted residual gastric volume exceeded one-half. Laparoscopic surgery was performed in 11 patients, while only 1 patient underwent open surgery. The mean duration of mToupet-like anastomosis was 48.3±8.7 minutes with an estimated intraoperative blood loss was 53.0±11.2 ml. All the 12 patients successfully achieved R0 resection. Among these patietns, the median postoperative hospital stay was 8.5 (7.0, 11.0) days, and the average hospitalization cost was 5.0±0.2 ten thousand yuan. No Clavien-Dindo grade II or higher complications were observed during the perioperative period. Patients were followed up for 6 to 8 months after operation, and no cases of reflux esophagitis were detected by gastroscopy, and no patient required long-term oral proton pump inhibitors.Conclusions:mToupet-like anastomosis for digestive tract reconstruction after proximal gastrectomy is a safe and feasible technique, demonstrating favorable preliminary efficacy.

Objective To investigate the expression and roles of serine/threonine and tyrosine kinases(TTK)and mouse dual microbody 2(MDM2)in cervical lesion tissue.Methods Patients(n=180)who underwent gynecological surgery at the First Affiliated Hospital of Henan University of Science and Technology from June 2022 to January 2024 were evaluated,including 50 patients in the cervical cancer group,50 patients in the high-grade lesion group,40 patients in the low-grade lesion group,and 40 patients in the non-cervical lesion(control)group.The mRNA expression and protein levels of TTK and MDM2 in each group were detected and compared by real-time quantitative PCR and immunohistochemistry,respectively.The diagnostic value of the mRNA expression levels of TTK and MDM2 in cer-vical cancer was analyzed using receiver operating characteristic curves.Results The mRNA levels of TTK and MDM2 significantly dif-fered among the four groups(P＜0.05);with increases in the lesion grade,the mRNA levels of TTK and MDM2 increased gradually and showed significant differences(P＜0.05).Immunohistochemical results showed that TTK was mainly located in the cytoplasm,whereas MDM2 was mainly located in the nucleus,with positive expression rates in cervical cancer tissues of 84.00％and 90.00％,respectively.These values were higher than those in other pathological tissues(P＜0.05).The mRNA levels of TTK and MDM2 were significantly cor-related with the degree of differentiation and lymph node metastasis(P＜0.05).The receiver operating characteristic curve showed that the areas under the curve of TTK and MDM2 mRNA for diagnosing cervical cancer were 0.803 and 0.798,respectively.Additionally,the area under the curve of the combined diagnosis was 0.912,which was higher than that observed for single detection(Z=2.230,2.553,P＜0.05).Conclusion The mRNA expression of TTK and MDM2 and their corresponding protein levels were increased in cervical lesions,demonstrating their potential as biological indicators for the clinical diagnosis of cervical cancer.

Objective:This study aimed to identify potential prognostic and biotherapeutic targets for oral squamous cell carcinoma(OSCC)by investigating the regulatory effects of M1/M2 macrophages on OSCC progression and their underlying mechanisms.Methods:The ef-fects of M1/M2 macrophages on the proliferation,migration,invasion,and apoptosis of SCC-15 cells were examined.Immunohistochemical staining was used to detect the expression of CD68 and M2 macrophage marker proteins in human OSCC tissue microarrays,and their cor-relations with clinicopathological features were analyzed.Results:M1 macrophages promoted apoptosis of SCC-15 cells while inhibiting their proliferation,migration,and invasion.In contrast,M2 macrophages exhibited the opposite effects.The infiltration density of tumor-associ-ated macrophages(TAMs)in OSCC tissues was significantly positively correlated(P<0.05)with lymph node metastasis.Conclusions:M1 mac-rophages suppress OSCC progression,whereas M2 macrophages promote it.