Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

Specialist nurses play an increasingly important role in clinical,and at the same time,they(especially enterostomal therapists/wound stoma specialist nurses)are often troubled by ethical problems when facing patients independently.Based on the"structured analysis form of clinical nursing ethics"constructed in the early stage,this paper used a case study method,took a chronic wound patient with postpartum fat liquefaction combined with thread-knot reaction as an example,conducted an in-depth analysis and reflection on the ethical problems of enterostomal therapists choosing treatment plans in clinical work,as well as proposed clinical recommendations.Base on this,this paper suggests that specialist nurses should fully consider the wishes and background characteristics of patients when nursing and treating patients,and use the four-box model of ethical analysis to analyze and solve ethical conflicts.

Objective:To explore the application effect of a multidisciplinary collaborative model led by stomatognathic therapists, utilizing a combination of platelet-rich plasma (PRP) and moist wound dressings in the comprehensive treatment of postoperative chronic wounds, and to provide references and guidance for improving the clinical practice and management level of patients with chronic wounds.Methods:This was a prospective randomized controlled trial. Eighty-eight patients with postoperative non-healing chronic wounds who visited the Chronic Wound Care Clinic of the Third Affiliated Hospital of Guangzhou Medical University from October 2018 to February 2023 were conveniently sampled. They were randomly divided into two groups using a coin toss method: the control group ( n = 41) received standard moist wound dressing therapy, while the experimental group ( n = 47) received comprehensive treatment based on PRP combined with moist wound dressings. This treatment, led by enterostoml therpist, involved the innovative nursing technique of simultaneous injection of PRP and coagulant mixture via a three-way connector and covering with ultra-thin foam dressing. The Pressure Ulcer Scale for Healing (PUSH) score, wound area healing index (WSHI), pain level during dressing change, wound healing time, and treatment satisfaction rate were observed in both groups. Results:Among 88 cases, there were 25 females and 16 males in the control group, aged 58.00 (37.00 ± 79.00) years old. There were 31 females and 16 males in the experimental group, aged 57.00 (35.00 ± 72.00) years old. The median PUSH scores of the experimental group on the 7th, 14th, and 21th day after wound treatment were 11.00, 9.00, and 7.00, respectively, which were better than those of the control group, which were 13.00, 11.00, and 9.00. The median WSHI scores were 0.35, 0.58, and 0.84 for the experimental group, respectively, which were better than those of the control group, which were 0.09, 0.30, and 0.50. The differences between the two groups were statistically significant ( Z values ranged from 4.08 to 8.20, all P<0.01). On the 7th, 14th, and 21st day of treatment, the pain scores of the experimental group were 3.00 (2.00,3.00), 2.00 (1.00, 2.00), 0.00 (0.00,1.00) points, respectively, which were lower than the 3.00 (3.00,3.50), 2.00 (2.00,3.00), and 2.00 (1.00, 2.00) points of the control group, and the differences were statistically significant ( Z values were 2.16, 3.38, 6.14, all P<0.05). The healing time in the experimental group was (37.04 ± 25.33) days, which was shorter than that in the control group, (52.88 ± 36.58) days, and the difference was statistically significant ( t = 5.53, P<0.05). The satisfaction rate in the experimental group was 97.87% (46/47), significantly higher than 87.80% (36/41) in the control group ( χ2 = 3.13, P<0.05). Conclusions:Under the leadership of stomatognathic therapists, the multidisciplinary collaborative model, employing comprehensive treatment techniques centered around PRP combined with moist wound dressings, demonstrated significant therapeutic efficacy for postoperative chronic wounds including shortening healing time, accelerating healing speed, reducing wound pain, and improving patient satisfaction, indicating high clinical application value and social benefits.

The structure of N-glycans on specific proteins can regulate innate and adaptive immunity via sensing environmental signals. Meanwhile, the structural diversity of N-glycans poses analytical challenges that limit the exploration of specific glycosylation functions. In this work, we used THP-1-derived macrophages as examples to show the vast potential of a N-glycan structural interpretation tool StrucGP in N-glycoproteomic analysis. The intact glycopeptides of macrophages were enriched and analyzed using mass spectrometry (MS)-based glycoproteomic approaches, followed by the large-scale mapping of site-specific glycan structures via StrucGP. Results revealed that bisected GlcNAc, core fucosylated, and sialylated glycans (e.g., HexNAc₄Hex₅Fuc₁Neu5Ac₁, N₄H₅F₁S₁) were increased in M1 and M2 macrophages, especially in the latter. The findings indicated that these structures may be closely related to macrophage polarization. In addition, a high level of glycosylated PD-L1 was observed in M1 macrophages, and the LacNAc moiety was detected at Asn-192 and Asn-200 of PD-L1, and Asn-200 contained Lewis epitopes. The precision structural interpretation of site-specific glycans and subsequent intervention of target glycoproteins and related glycosyltransferases are of great value for the development of new diagnostic and therapeutic approaches for different diseases.
Humans ; B7-H1 Antigen ; Glycosylation ; Polysaccharides/metabolism*

Stroke is the second leading cause of death worldwide,and oxidative stress plays a crucial role.Celastrol exhibits strong antioxidant properties in several diseases;however,whether it can affect oxidation in cerebral ischemic-reperfusion injury(CIRI)remains unclear.This study aimed to determine whether celastrol could reduce oxidative damage during CIRI and to elucidate the underlying mechanisms.Here,we found that celastrol attenuated oxidative injury in CIRI by upregulating nuclear factor E2-related factor 2(Nrf2).Using alkynyl-tagged celastrol and liquid chromatography-tandem mass spectrometry,we showed that celastrol directly bound to neuronally expressed developmentally downregulated 4(Nedd4)and then released Nrf2 from Nedd4 in astrocytes.Nedd4 promoted the degradation of Nrf2 through K48-linked ubiquitination and thus contributed to astrocytic reactive oxygen species production in CIRI,which was significantly blocked by celastrol.Furthermore,by inhibiting oxidative stress and astrocyte activation,celastrol effectively rescued neurons from axon damage and apoptosis.Our study uncovered Nedd4 as a direct target of celastrol,and that celastrol exerts an antioxidative effect on as-trocytes by inhibiting the interaction between Nedd4 and Nrf2 and reducing Nrf2 degradation in CIRI.

Objective:To apply the multi-modal deep learning model to automatically classify the ultra-widefield fluorescein angiography (UWFA) images of diabetic retinopathy (DR).Methods:A retrospective study. From 2015 to 2020, 798 images of 297 DR patients with 399 eyes who were admitted to Eye Center of Renmin Hospital of Wuhan University and were examined by UWFA were used as the training set and test set of the model. Among them, 119, 171, and 109 eyes had no retinopathy, non-proliferative DR (NPDR), and proliferative DR (PDR), respectively. Localization and assessment of fluorescein leakage and non-perfusion regions in early and late orthotopic images of UWFA in DR-affected eyes by jointly optimizing CycleGAN and a convolutional neural network (CNN) classifier, an image-level supervised deep learning model. The abnormal images with lesions were converted into normal images with lesions removed using the improved CycleGAN, and the difference images containing the lesion areas were obtained; the difference images were classified by the CNN classifier to obtain the prediction results. A five-fold cross-test was used to evaluate the classification accuracy of the model. Quantitative analysis of the marker area displayed by the differential images was performed to observe the correlation between the ischemia index and leakage index and the severity of DR.Results:The generated fake normal image basically removed all the lesion areas while retaining the normal vascular structure; the difference images intuitively revealed the distribution of biomarkers; the heat icon showed the leakage area, and the location was basically the same as the lesion area in the original image. The results of the five-fold cross-check showed that the average classification accuracy of the model was 0.983. Further quantitative analysis of the marker area showed that the ischemia index and leakage index were significantly positively correlated with the severity of DR ( β=6.088, 10.850; P<0.001). Conclusion:The constructed multimodal joint optimization model can accurately classify NPDR and PDR and precisely locate potential biomarkers.

Objective To summarize the prenatal ultrasonographic characteristics of anomalous origin of one pulmonary artery from the aorta( AOPA ) ,and describe the diagnostic and clinical outcomes of fetal AOPA . Methods Echocardiographic characteristics of 3 fetuses with AOPA were reviewed . The ultrasonographic features were comparatively analyzed with postpartum autopsy findings . The relevant literature were reviewed and the experience of prenatal diagnosis of AOPA were summarized . Results In 3 cases with AOPA ,2 cases were the proximal type and anomalous origin of right pulmonary artery from the aorta ,of them ,one was accompanied with coarctation of the aorta , the other was accompanied with aortopulmonary window . One case was the distal type and anomalous origin of left pulmonary artery from the left innominate aorta ,it was only associated with mirror right aortic arch and right ductus arteriosus . Ultrasound characteristics were no bifurcation in distal pulmonary artery ,main trunk of pulmonary artery extends directly to one branch ,the other branch originated from the ascending aorta or left innominate artery . Three cases were confirmed by postpartum autopsy . Conclusions There are some characteristic signs on ultrasonic features of AOPA . Prenatal ultrasound has important value for diagnosis of AOPA .