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.

BACKGROUND:Mangiferin is a biphenylpyridone compound extracted from mango leaves,bark and roots.Previous studies have shown that mangiferin can exert anti-systemic inflammatory effects through the activation of transcription factors such as NF-κB and JAK/STAT. OBJECTIVE:To investigate the effects and mechanisms of mangiferin on proliferation,migration and inflammatory factor release of rheumatoid arthritis fibroblast-like synovial cells(RA-FLS). METHODS:RA-FLS were divided into blank group,R848(TLR7/8 agonists)stimulated group,mangiferin low-,medium-,high-dose groups(2,4 and 8 μg/mL)and positive control group(Cu-CPT8,TLR8 pathway inhibitor).The cytotoxic effect of different mass concentrations of mangiferin was detected using cell counting kit-8 method and the final cellular dosing mass concentration was screened.The proliferation ability of RA-FLS was detected by cell clone formation assay,the migration ability of RA-FLS was detected by scratch assay and Transwell migration assay,and the expression of interleukin 1β,interleukin 6 and tumor necrosis factor α mRNA in RA-FLS was detected by qRT-PCR. RESULTS AND CONCLUSION:Compared with the blank group,the viability of RA-FLS was inhibited after treatment with mangiferin at 2-10 μg/mL,but there was no significant difference among groups(P>0.05),indicating that the toxic effect on RA-FLS was minimal.Compared with the R848-stimulated group,mangiferin decreased the number of cell clones,the scratch healing rate and the number of migrating cells in all dosing groups(P<0.01);and the expression of interleukin 1β,interleukin 6 and tumor necrosis factor α mRNA was also reduced in the mangostin medium-and high-dose groups(P<0.01).Compared with the R848-stimulated group,the number of cell clones,the scratch healing rate and the number of migrating cells as well as the expression levels of interleukin 6 and tumor necrosis factor α mRNA were significantly reduced in the positive control group(P<0.05,P<0.01).But there was no significant difference in the expression level of interleukin 1β.To conclude,mangiferin may exert its anti-rheumatoid arthritis effects through the TLR7/8 signaling pathway by inhibiting RA-FLS proliferation,migration,and inflammatory factor release.

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.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.

Objective:Using intrapartum ultrasound monitoring to observe and analyse the impact of upright sit-ting delivery on fetal head descent and maternal and infant outcomes during the second stage of labor.Methods:This study used a prospective randomized controlled study,including 110 pregnant women with full-term and com-plete opening of the cervix who were able to undergo vaginal delivery at Beijing Obstetrics and Gynecology Hospi-tal,Capital Medical University from February to September 2023.They were randomly divided into a control group(n=56)and an upright group(n=54)by simple randomization.The control group was delivered in routine supine or semi-supine positions,and the upright group was delivered in upright positions.The parturients in both groups were examined by color Doppler ultrasound at the beginning of the second stage of labor when the cervix was full-y opened to guide delivery and 30 minutes after guidance of the second stage of labor.Abdominal and perineal two-dimensional ultrasound was used to detect fetal head angle of progression(AOP),fetal head perineal dis-tance(HPD),and fetal orientation.At the same time,the time of the second stage of labor and delivery outcome were recorded.The changes in AOP,angle and position of HPD,delivery duration,and maternal and fetal out-comes between the two groups were analyzed.Results:At the beginning of the second stage of labor,there was no significant difference in AOP,HPD,and the rate of non-occipital anterior position of fetal position between the two groups(P＞0.05).After 30 minutes of guiding delivery,compared with the control group,the AOP angle in-creased,the HPD distance shortened,and the proportion of non-occipital anterior fetal position was reduced in the upright position group,and all the differences were statistically significant(P＜0.05).At the same time,compared with the control group,the spontaneous delivery rate,the duration of the second stage of labor,forceps delivery rate,lateral perineal incision rate,perineal edema rate,and oxytocin use rate were significantly decreased in the upright position group,and all the differences were statistically significant(P＜0.05).All newborns were born a-live,and the 1-minute Apgar score was 10 for all newborns in both groups.There was no significant difference in birth weight between the two groups(P＞0.05).Conclusions:When the parturient delivers in an upright sitting position in the second stage of labor,the AOP increases,the HPD shortens,and the time of the second stage of labor is accelerated to increase the natural delivery rate,reduce medical intervention,and reduce maternal and in-fant complications during delivery.