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.

Objective To observe the repair effect and mechanism of Danhuang Powder-containing serum on high glucose-induced vascular endothelial cell injury.Methods Danhuang Powder-containing serum was prepared.Human umbilical vein endothelial cells(HUVECs)were cultured to be divided into control group,recombinant human epidermal growth factor(called"growth factor"for short)group,Danhuang Powder group,high glucose group,high glucose+growth factor group,and high glucose+Danhuang Powder group.After corresponding intervention in each group for 48 hours,the cell ultrastructure and autophagy were observed under transmission electron microscope,apoptosis was detected by flow cytometry,and the protein expression levels of vascular endothelial growth factor(VEGF),epidermal growth factor(EGF)and basic fibroblast growth factor(bFGF)in the cells were detected by Western Blot.Results(1)The intra-mitochondrial ridges in the control group were clearly visible,autophagosomes and autolysosomes were fewer;mitochondria in the high glucose group were swollen and irregular,and appeared vacuolated;and the more typical autophagy-like structures were seen in the high glucose+Danhuang Powder group.(2)Compared with the high glucose group and high glucose+growth factor group,the apoptosis rate of cells in the high glucose+Danhuang Powder group was significantly decreased(P<0.05).(3)Compared with the high glucose group and the high glucose+growth factor group,the protein expression levels of VEGF,EGF and bFGF in the cells of the high glucose+Danhuang Powder group were significantly increased(P<0.05).Conclusion Danhuang Powder-containing serum can reduce the high glucose-induced damage in HUVEC cells,and its mechanism may be related to the activation of mitochondrial autophagy,and the inhibition of apoptosis,as well as the up-regulation of the expression of VEGF,EGF and bFGF.

Improving the sense of belonging of the elderly in nursing institutions is of great significance to enhance their sense of self-worth and self-identity,and further meet the spiritual needs of the elderly in nursing institutions.This paper reviewed the concept,current situation,evaluation tools and influencing factors of the elderly's sense of belonging in nursing institutions,and proposes that there is a lack of tools to measure the sense of belonging of the elderly in nursing institutions,and the intervention strategies with caregivers as the core needs to be further explored.

The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.

Objective:To investigate the clinical efficacy of different surgical approaches for repairing eyelid coloboma.Methods:Patients with the divided nevus of eyelid treated at Hypospadias Plastic Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2005 to January 2022 were included. The surgeries were categorized into 4 types. (1) Direct excision and suture. (2) Local skin grafts: covering the defect with split- or full-thickness skin grafts according to the size of the defect on the upper and lower eyelids. (3) Combined skin grafts with orbicularis oculi myocutaneous flap: grafting skin flaps for defects on the upper eyelid and near the lower eyelid, and temporal area skin flaps based on the same side orbicularis oculi muscle pedicle for lower eyelid defects. (4) Temporal area expanded flap based on the orbicularis oculi muscle combined with skin grafts: the surgery was divided into two stages, the first stage involves the placement of an expander in the temporal area of the affected side, and the second stage involves the removal of the expander, excision of eyelid lesion tissue and formation of an island-shaped skin flap with the orbital part of the orbicularis oculi muscle pedicle as the pedicle, which was rotated 180° to cover the lower eyelid defect. Defects near the upper and lower eyelid margins were still covered with skin grafts. Follow-up was conducted through outpatient visits, telephone calls, and WeChat messaging to assess facial appearance postoperatively. SPSS 22.0 statistical software was used for analysis, and the measurement data were expressed as Mean±SD, the count data were expressed as percentage, and the comparison of preoperative and postoperative was calculated by aesthetic and functional status of facial soft-tissue deformities (A&F scores) within the group was performed by paired t-test, the difference was considered statistically significant at P<0.05. Results:A total of 34 patients were included; average age was 17.7±15.3 years. The size of the lesions varied from the smallest nevus measuring 0.3 cm × 0.2 cm and the largest measuring 14.0 cm × 14.0 cm. Direct excision with suturing was performed in 6 cases, among which 2 cases received simultaneous double eyelid surgery, the postoperative A&F score (4.54 ± 1.32) was higher than the preoperative (3.28 ± 0.98) score, the difference was not statistically significant ( P>0.05). The skin graft was performed in 10 cases, the postoperative A&F score (5.13 ± 1.59) was higher than the preoperative (2.25 ± 1.59), the difference was not statistically significant ( P>0.05). The skin graft combined with using orbicularis oculi muscle skin flap was performed in 7 cases, the postoperative A&F score of our patients (5.54 ±1.46) was significantly higher than the preoperative (2.18 ±1.61 ), the difference was statistically significant ( P<0.05). The skin graft combined with the expanded temporal area flap based on the orbicularis oculi muscle was performed in 11 cases, the A&F score of our patients after repair (4.95 ±0.60) was improved compared with the preoperative (2.18±1.48) score, and the difference was not statistically significant ( P>0.05). Twenty-eight patients were followed up for 8 to 81 months while 6 cases lost due to change of contact information, among whom three developed secondary ectropion of the lower eyelid, three experienced recurrence of nevi in the incision and grafting area, two exhibited significant pigmentation in the grafting area, and one developed secondary ptosis of the upper eyelid, while the rest of the patients were satisfied with the repair results. Conclusion:Surgical excision is the only effective treatment for eyelid cleft nevi with different clinical presentations. Covering defects on the upper and lower eyelids with different tissue transplantation method can disperse the entire nevus, resulting in more stable long-term repair effects and a more natural appearance. The combination of an orbicularis muscle flap and a free skin graft provides more stable result in the plastic surgery of medium to large eyelid split nevi. Free skin grafting of the upper and lower lids near the margins, and flap grafting of the lower lids are more consistent with the physiological state of the upper and lower lids, and the combination of different tissue grafting method can make the appearance of the face more reasonable.

Indocyanine green ( ICG ) is a water-soluble biological fluorescent contrast agent with infrared absorption and emission characteristics. Indocyanine green angiography ( ICGA ) takes advantage of this biological property. After ICG is injected into the vascular system, the blood flow dynamic map is captured by high-speed photography or real-time photography, which can realize the visualization of vascular structure and tissue blood supply. Currently, ICGA is the only technique for intraoperative quantitative evaluation of flap blood perfusion, and it is widely used in plastic surgery and microsurgery. According to the relevant literature reports at home and abroad in recent years, this review introduced the working principle and method of ICGA and its research progress in evaluating the blood perfusion of pedicled skin flaps. It is considered that this technique still has some limitations in methodology and accuracy of result, and it is still necessary to combine the clinical experience of doctors in the evaluation of tissue perfusion of pedicled skin flaps.

Objective:To investigate the clinical efficacy of different surgical approaches for repairing eyelid coloboma.Methods:Patients with the divided nevus of eyelid treated at Hypospadias Plastic Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2005 to January 2022 were included. The surgeries were categorized into 4 types. (1) Direct excision and suture. (2) Local skin grafts: covering the defect with split- or full-thickness skin grafts according to the size of the defect on the upper and lower eyelids. (3) Combined skin grafts with orbicularis oculi myocutaneous flap: grafting skin flaps for defects on the upper eyelid and near the lower eyelid, and temporal area skin flaps based on the same side orbicularis oculi muscle pedicle for lower eyelid defects. (4) Temporal area expanded flap based on the orbicularis oculi muscle combined with skin grafts: the surgery was divided into two stages, the first stage involves the placement of an expander in the temporal area of the affected side, and the second stage involves the removal of the expander, excision of eyelid lesion tissue and formation of an island-shaped skin flap with the orbital part of the orbicularis oculi muscle pedicle as the pedicle, which was rotated 180° to cover the lower eyelid defect. Defects near the upper and lower eyelid margins were still covered with skin grafts. Follow-up was conducted through outpatient visits, telephone calls, and WeChat messaging to assess facial appearance postoperatively. SPSS 22.0 statistical software was used for analysis, and the measurement data were expressed as Mean±SD, the count data were expressed as percentage, and the comparison of preoperative and postoperative was calculated by aesthetic and functional status of facial soft-tissue deformities (A&F scores) within the group was performed by paired t-test, the difference was considered statistically significant at P<0.05. Results:A total of 34 patients were included; average age was 17.7±15.3 years. The size of the lesions varied from the smallest nevus measuring 0.3 cm × 0.2 cm and the largest measuring 14.0 cm × 14.0 cm. Direct excision with suturing was performed in 6 cases, among which 2 cases received simultaneous double eyelid surgery, the postoperative A&F score (4.54 ± 1.32) was higher than the preoperative (3.28 ± 0.98) score, the difference was not statistically significant ( P>0.05). The skin graft was performed in 10 cases, the postoperative A&F score (5.13 ± 1.59) was higher than the preoperative (2.25 ± 1.59), the difference was not statistically significant ( P>0.05). The skin graft combined with using orbicularis oculi muscle skin flap was performed in 7 cases, the postoperative A&F score of our patients (5.54 ±1.46) was significantly higher than the preoperative (2.18 ±1.61 ), the difference was statistically significant ( P<0.05). The skin graft combined with the expanded temporal area flap based on the orbicularis oculi muscle was performed in 11 cases, the A&F score of our patients after repair (4.95 ±0.60) was improved compared with the preoperative (2.18±1.48) score, and the difference was not statistically significant ( P>0.05). Twenty-eight patients were followed up for 8 to 81 months while 6 cases lost due to change of contact information, among whom three developed secondary ectropion of the lower eyelid, three experienced recurrence of nevi in the incision and grafting area, two exhibited significant pigmentation in the grafting area, and one developed secondary ptosis of the upper eyelid, while the rest of the patients were satisfied with the repair results. Conclusion:Surgical excision is the only effective treatment for eyelid cleft nevi with different clinical presentations. Covering defects on the upper and lower eyelids with different tissue transplantation method can disperse the entire nevus, resulting in more stable long-term repair effects and a more natural appearance. The combination of an orbicularis muscle flap and a free skin graft provides more stable result in the plastic surgery of medium to large eyelid split nevi. Free skin grafting of the upper and lower lids near the margins, and flap grafting of the lower lids are more consistent with the physiological state of the upper and lower lids, and the combination of different tissue grafting method can make the appearance of the face more reasonable.

Indocyanine green ( ICG ) is a water-soluble biological fluorescent contrast agent with infrared absorption and emission characteristics. Indocyanine green angiography ( ICGA ) takes advantage of this biological property. After ICG is injected into the vascular system, the blood flow dynamic map is captured by high-speed photography or real-time photography, which can realize the visualization of vascular structure and tissue blood supply. Currently, ICGA is the only technique for intraoperative quantitative evaluation of flap blood perfusion, and it is widely used in plastic surgery and microsurgery. According to the relevant literature reports at home and abroad in recent years, this review introduced the working principle and method of ICGA and its research progress in evaluating the blood perfusion of pedicled skin flaps. It is considered that this technique still has some limitations in methodology and accuracy of result, and it is still necessary to combine the clinical experience of doctors in the evaluation of tissue perfusion of pedicled skin flaps.

Objective:Non-alcoholic fatty liver disease(NAFLD)is a common metabolic disorder in overweight and obese children,and its etiology and pathogenesis remain unclear,lacking effective preventive and therapeutic measures.This study aims to explore the association between whole blood copper,zinc,calcium,magnesium and iron levels and NAFLD in overweight and obese children aged 6 to 17 years,providing a scientific basis for the prevention and intervention of early NAFLD in overweight and obese children. Methods:A cross-sectional study design was used to collect relevant data from overweight and obese children who visited the Hunan Children's Hospital from January 2019 to December 2021 through questionnaire surveys.Fasting blood samples were collected from the subjects,and various indicators such as blood glucose,blood lipid,and mineral elements were detected.All children were divided into an overweight group(n=400)and a NAFLD group(n=202).The NAFLD group was divided into 2 subgroups according to the ALT level:A non-alcoholic fatty liver(NAFL)group and a non-alcoholic steatohepatitis(NASH)group.Logistic regression analysis was used to analyze the association between minerals(copper,zinc,calcium,magnesium,and iron)and NAFLD,NAFL and NASH. Results:A total of 602 subjects were included,of whom 73.6%were male,with a median age of 10(9,11)years,and a body mass index(BMI)of 24.9(22.7,27.4)kg/m2.The intergroup comparison results showed that compared with the overweight group,the NAFLD group had higher levels of age,BMI,diastolic blood pressure(DBP),systolic blood pressure(SBP),triglyceride(TG),low density lipoprotein(LDL),alanine transaminase(ALT)and aspartate aminotransferase(AST),and lower level of high density lipoprotein(HDL).The NAFL group had higher levels of age,BMI,DBP,SBP,ALT,and AST,and lower levels of HDL compared with the overweight group.The levels of age,BMI,DBP,SBP,TG,LDL,ALT,and AST of NASH were higher than those in the overweight group,while the level of HDL was lower than that in overweight group(all P<0.017).After adjusting for a variety of confounders,the OR of NAFLD for the highest quantile of iron was 1.79(95%CI 1.07 to 3.00)compared to the lowest quantile,and no significant association was observed between copper,zinc,calcium,and magnesium,and NAFLD.The subgroup analysis of NAFLD showed that the OR for the highest quantile of iron in children with NAFL was 2.21(95%CI 1.26 to 3.88),while no significant association was observed between iron level and NASH.In addition,no significant associations were observed between copper,zinc,calcium,and magnesium levels and NAFL or NASH. Conclusion:High iron level increases the risk of NAFLD(more likely NAFL)in overweight and obese children,while copper,zinc,calcium,magnesium,and other elements are not associated with the risk of NAFLD in overweight and obese children.

OBJECTIVES: The increasing costs of hospital delivery have increased the economic burden of pregnant women, and the mode of delivery is the main factor affecting the costs of hospital delivery. This study aims to explore the difference in costs between cesarean section and natural delivery, and to provide reference for controlling the increase of hospital delivery costs. METHODS: The data of inpatient delivery in the Hunan Maternal and Child Health Care Hospital from January 2016 to December 2020 were selected to compare the total inpatient costs and average daily costs of cesarean section and natural delivery. The linear trend model was used to analyze the trend change of inpatient delivery costs and the generalized linear model was used to analyze the influential factors for inpatient delivery costs. RESULTS: The average hospitalization costs of cesarean section (10 447.25 yuan) were higher than that of natural delivery (5 567.95 yuan), and the average daily costs of cesarean section (1 902.57 yuan) were higher than those of natural delivery (1 666.40 yuan). There was no significant increase or decrease in trend for cesarean section, while the average annual growth rate of the costs of natural delivery was 11.79%. The main factors affecting the hospitalization costs of cesarean section and natural delivery included age, occupation, medical insurance, route of admission, length of stay, premature delivery and complications (all P<0.05). CONCLUSIONS The total hospitalization costs and average daily costs of cesarean section are higher than those of natural delivery, but the costs of natural delivery show a faster growth trend, and the hospitalization costs of cesarean section and natural delivery should be controlled by targeted measures.
Child ; Female ; Pregnancy ; Humans ; Cesarean Section ; Hospitalization ; Hospitals ; Hospital Costs ; Inpatients ; Retrospective Studies