Objective:To study the effects and mechanisms of activation of human lung fibroblasts (MRC-5) by exosomal RNA hsa _ circ _ 0006357 (circEZH2) derived from non-small cell lung cancer.Methods:Western blot was used to detect exosome molecular markers, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and cell invasion assays to detect the effect of non-small cell lung cancer-derived exosomes on MRC-5 activation. A circRNA microarray analysis was performed in serum exosomes from patients with non-small cell lung cancer (collected at Ningbo University People's Hospital, September 2023), and levels of circEZH2 were measured in serum exosomes from non-small cell lung cancer by RT-qPCR analysis. The effects of circEZH2 on MRC-5 activation were explored using wound healing assays, Transwell assays, RT-qPCR, cellular immunofluorescence, and western blot. The regulatory effect of circEZH2 on miR-495-3p/TPD52 axis and NF-κB pathway through dual-luciferase assay, immunofluorescence, and western blot.Results:Exosomes derived from non-small cell lung cancer cells were shown to promote MRC-5 cell invasiveness, the number of cells invaded in co-culture with exosomes derived from normal human bronchial epithelial cells was (42±5), and the number of cells invaded in co-culture with exosomes derived from non-small cell lung cancer cells (SPC-A1, H1299, A549 cells) was (246±7), (89±4), (69±14), expression of markers of fibroblast activation (α-SMA, FAP), and cytokines (IL-6, IL-8, P＜0.05). CircEZH2 expression was significantly upregulated in the serum exosomes of non-small cell lung cancer ( P＜0.01). Alternatively, co-culture of exosomes derived from non-small cell lung cancer cells with MRC-5 cells promoted circEZH2 expression ( P＜0.05). Functionally, overexpression of circEZH2 promoted MRC-5 cell migration and invasion [the cell migration rates were (30.81±2.54)% and (60.29±8.34)%, respectively, and the cell invasion numbers were (48.00±13.58) and (115.00±9.50), respectively, P＜0.05]. RT-qPCR, western blot, and immunofluorescence assays demonstrated a significant increase in the expression of the pro-inflammatory genes IL-6 and IL-8 in MRC-5 cells as well as the activation markers α-SMA and FAP in fibroblasts ( P＜0.05) following the expression of circEZH2. Mechanistically, circEZH2 may function as ceRNA to regulate miR-495-3p, promote the expression of TPD52, and activate the NF-κB pathway to promote the activation of MRC-5. Conclusion:Exosomal circEZH2, derived from non-small cell lung cancer, may promote the activation of fibroblasts by activating the NF-κB pathway through the miR-495-3p/TPD52 axis.

Objective To investigate the clinical manifestations,molecular genetics and gonadal pathol-ogy characteristics of patients with disorders of sex development(DSD),and to summarize the clinical experi-ence of identifying rare diseases from common symptoms.Methods The clinical data of 416 patients with DSD diagnosed and treated in the multidisciplinary center of DSD of Guangzhou Women and Children's Medical Cen-ter from May 2018 to August 2023 were retrospectively analyzed,summarized and discussed.Results Accord-ing to chromosome karyotype,416 cases of DSD were classified into three types:92 cases(22.1％)of abnormal sex chromosome karyotype,285 cases(68.5％)of 46,XY karyotype and 39 cases(9.4％)of 46,XX karyotype.Among the 92 patients with abnormal sex chromosome karyotype,59 cases were raised as males,18 cases(30.5％)complained of short penis with hypospadias and cryptorchidism.The most common karyotype was 45,X/46,XY(58 cases,63.0％).Among the 285 patients with 46,XY karyotype,238 cases were raised as males,and 63 cases(26.5％)complained of short penis and hypospadias;47 cases were raised as females,and 13 ca-ses(27.7％)complained of inguinal mass.A total of 216 patients with 46,XY karyotype were subjected to whole exome gene detection,and 155 cases(71.8％)were found to have molecular pathogenesis with the clinical phe-notype.Among the 39 patients with 46,XX karyotype,19 cases were raised as males,and 8 cases(42.1％)com-plained of short penis and hypospadias.In the 18 cases of gonad biopsy,17 cases showed testicular tissue in go-nads.Whole exome sequencing was performed in 14 cases.NR5A1 gene heterozygous mutation,SRY gene muta-tion and SOX3 gene mutation were found in 2 cases,respectively(14.3％).Twenty cases were raised as females,and 14 cases(70.0％)complained of clitoral hypertrophy.Gonad biopsy was performed in 8 cases,with 7 cases of ovotestis(87.5％)and 1 case of NR5A1 gene heterozygous mutation(14.3％).Conclusions The etiologies of DSD are complex and diverse,and the clinical manifestations are various,which can be manifested as hypospa-dias,micropenis,cryptorchidism and other common symptoms of the urinary system.Different etiologies have dif-ferent treatment options.Therefore,chromosome karyotype,molecular genetic testing and gonadal pathology can be used to clarify the cause of disease,especially for rare diseases,improve the detection rate,reduce the rate of missed diagnosis,and ensure reasonable treatment,especially sex selection.

Background::Cerebrovascular disease (CVD) ranks among the foremost factors responsible for mortality on a global scale. The mortality patterns of CVDs and temporal trends in China need to be well-illustrated and updated.Methods::We collected mortality data on patients with CVD from Chinese Center for Disease Control and Prevention’s Disease Surveillance Points (CDC-DSP) system. The mortality of CVD in 2020 was described by age, sex, residence, and region. The temporal trend from 2013 to 2019 was evaluated using joinpoint regression, and estimated rates of decline were extrapolated until 2030 using time series models.Results::In 2019, the age-standardized mortality in China (ASMRC) per 100,000 individuals was 113.2. The ASMRC for males (137.7/10 5) and rural areas (123.0/10 5) were both higher when stratified by gender and urban/rural residence. The central region had the highest mortality (126.5/10 5), the western region had a slightly lower mortality (123.5/10 5), and the eastern region had the lowest mortality (97.3/10 5). The age-specific mortality showed an accelerated upward trend from aged 55-59 years, with maximum mortality observed in individuals over 85 years of age. The age-standardized mortality of CVD decreased by 2.43% (95% confidence interval, 1.02-3.81%) annually from 2013 to 2019. Notably, the age-specific mortality of CVD increased from 2013 to 2019 for the age group of over 85 years. In 2020, both the absolute number of CVD cases and the crude mortality of CVD have increased compared to their values in 2019. The estimated total deaths due to CVD were estimated to reach 2.3 million in 2025 and 2.4 million in 2030. Conclusion::The heightened focus on the burden of CVD among males, rural areas, the central and western of China, and individuals aged 75 years and above has emerged as a pivotal determinant in further decreasing mortalities, consequently presenting novel challenges to strategies for disease prevention and control.

Objective:To study the effects and mechanisms of activation of human lung fibroblasts (MRC-5) by exosomal RNA hsa _ circ _ 0006357 (circEZH2) derived from non-small cell lung cancer.Methods:Western blot was used to detect exosome molecular markers, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and cell invasion assays to detect the effect of non-small cell lung cancer-derived exosomes on MRC-5 activation. A circRNA microarray analysis was performed in serum exosomes from patients with non-small cell lung cancer (collected at Ningbo University People's Hospital, September 2023), and levels of circEZH2 were measured in serum exosomes from non-small cell lung cancer by RT-qPCR analysis. The effects of circEZH2 on MRC-5 activation were explored using wound healing assays, Transwell assays, RT-qPCR, cellular immunofluorescence, and western blot. The regulatory effect of circEZH2 on miR-495-3p/TPD52 axis and NF-κB pathway through dual-luciferase assay, immunofluorescence, and western blot.Results:Exosomes derived from non-small cell lung cancer cells were shown to promote MRC-5 cell invasiveness, the number of cells invaded in co-culture with exosomes derived from normal human bronchial epithelial cells was (42±5), and the number of cells invaded in co-culture with exosomes derived from non-small cell lung cancer cells (SPC-A1, H1299, A549 cells) was (246±7), (89±4), (69±14), expression of markers of fibroblast activation (α-SMA, FAP), and cytokines (IL-6, IL-8, P＜0.05). CircEZH2 expression was significantly upregulated in the serum exosomes of non-small cell lung cancer ( P＜0.01). Alternatively, co-culture of exosomes derived from non-small cell lung cancer cells with MRC-5 cells promoted circEZH2 expression ( P＜0.05). Functionally, overexpression of circEZH2 promoted MRC-5 cell migration and invasion [the cell migration rates were (30.81±2.54)% and (60.29±8.34)%, respectively, and the cell invasion numbers were (48.00±13.58) and (115.00±9.50), respectively, P＜0.05]. RT-qPCR, western blot, and immunofluorescence assays demonstrated a significant increase in the expression of the pro-inflammatory genes IL-6 and IL-8 in MRC-5 cells as well as the activation markers α-SMA and FAP in fibroblasts ( P＜0.05) following the expression of circEZH2. Mechanistically, circEZH2 may function as ceRNA to regulate miR-495-3p, promote the expression of TPD52, and activate the NF-κB pathway to promote the activation of MRC-5. Conclusion:Exosomal circEZH2, derived from non-small cell lung cancer, may promote the activation of fibroblasts by activating the NF-κB pathway through the miR-495-3p/TPD52 axis.

Serine/arginine-rich splicing factor 7 (SRSF7), a known splicing factor, has been revealed to play oncogenic roles in multiple cancers. However, the mechanisms underlying its oncogenic roles have not been well addressed. Here, based on N⁶-methyladenosine (m⁶A) co-methylation network analysis across diverse cell lines, we find that the gene expression of SRSF7 is positively correlated with glioblastoma (GBM) cell-specific m⁶A methylation. We then indicate that SRSF7 is a novel m⁶A regulator, which specifically facilitates the m⁶A methylation near its binding sites on the mRNAs involved in cell proliferation and migration, through recruiting the methyltransferase complex. Moreover, SRSF7 promotes the proliferation and migration of GBM cells largely dependent on the presence of the m⁶A methyltransferase. The two m⁶A sites on the mRNA for PDZ-binding kinase (PBK) are regulated by SRSF7 and partially mediate the effects of SRSF7 in GBM cells through recognition by insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). Together, our discovery reveals a novel role of SRSF7 in regulating m⁶A and validates the presence and functional importance of temporal- and spatial-specific regulation of m⁶A mediated by RNA-binding proteins (RBPs).
Humans ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Glioblastoma/genetics* ; Methyltransferases/metabolism* ; RNA Splicing Factors/metabolism* ; RNA, Messenger/genetics* ; RNA-Binding Proteins/metabolism* ; Serine-Arginine Splicing Factors/metabolism* ; RNA Methylation/genetics*

Objective: To understand the status quo and influencing factors of overweight and obesity in preschool children, and to provide scientific theoretical reference for the prevention and control of overweight and obesity in preschool children in Urumqi. Methods: Stratified cluster sampling method was adopted to select 1 897 preschool children from 10 kindergartens in Urumqi from October to December in 2021 to understand the status quo of overweight and obesity of preschool children by measuring their height and weight. The influencing factors were collected by questionnaire survey, including Chinese preschoolers eating behavior questionnaire, Chinese preschooler s caregivers feeding behavior scale, 3-6 year old children s home nurture environment scale, and characteristics and influencing factors of physical activity among preschool children. Results: The prevalence of overweight and obesity in preschool children was 31.21% (592), including 19.50% (370) overweight and 11.70% (222) obesity.Childhood overweight and obesity detection rates varied significantly by age, sex, child dietary habits, father BMI, maternal BMI, and maternal pre pregnancy BMI ( χ 2=19.63,28.75,9.45,18.21,18.45,19.36, P <0.05). Multivariate Logistic regression analysis showed that gender, children s eating habits, paternal BMI, pregnancy BMI, satiety responsiveness, external eating, initiative eating, weight concerns, behavior restricted feeding, physical activity and family physical activity environment were the influencing factors of overweight and obesity in preschool children( OR =0.52,1.43,1.51,1.44,0.69,0.74,1.35,1.71,0.81,0.96,1.10, P < 0.05 ). Conclusion Overweight and obesity in preschool children are popular in Urumqi. Education, diet control and scientific exercise should be strengthened to prevent childhood overweight and obesity.

This study sought to assess the therapeutic effect of celecoxib (CEL)-loaded polylactic acid-glycolic acid copolymer (PLGA) microspheres on rheumatoid arthritis in rats after intra-articular injection.The celecoxib-loaded microspheres (CEL-MS) were prepared by the O/W solvent volatilization method with PLGA as carrier.In order to investigate the therapeutic effect of CEL-MS on rheumatoid arthritis in rats after intra-articular injection, a rat model of adjuvant arthritis (AA) was constructed by complete Freund''s adjuvant, and the evaluation indicators of the therapeutic effect were rat paw swelling, arthritis index,spleen index and joint synovial histopathological examination. The results showed that the microspheres had a smooth spherical morphology with a particle size of (2.1 ± 0.3) μm and a drug loading efficiency of (20.8 ± 0.6)%.The results of the in vivo efficacy test showed that intra-articular injection of CEL-MS compared to the CEL suspension oral and the celecoxib suspension intra-articular injection in adjuvant arthritis rat model can significantly reduce joint swelling and arthritis index, thus effectively inhibiting synovial inflammation.The above results indicate that intra-articular injection of CEL-MS has a good therapeutic effect on rheumatoid arthritis in rats.

Objective:To investigate the clinical significance of fluid-attenuated inversion recovery (FLAIR) vascular hyperintensities (FVHs) in patients with chronic atherosclerotic middle cerebral artery occlusion.Methods:From July 2016 to November 2020, patients admitted to the Department of Neurology, Jiangdu People's Hospital of Yangzhou and first found chronic atherosclerotic middle cerebral artery occlusion were enrolled. The demographic, clinical and MRI imaging data were collected. According to the presence or absence of acute cerebral infarction, they were divided into the non-acute cerebral infarction group and the acute cerebral infarction group. According to the modified Rankin Scale score at 3 months after the onset, patients with acute cerebral infarction were further divided into the good outcome group (≤2) and the poor outcome group (>2). A multivariate logistic regression model was used to analyze the independent correlation between FVHs and the risk of cerebral infarction in patients with chronic atherosclerotic middle cerebral artery occlusion and the outcome in patients with cerebral infarction. Results:A total of 94 patients with chronic atherosclerotic middle cerebral artery occlusion were enrolled, including 38 with non-acute cerebral infarction (40.4%) and 56 with acute cerebral infarction (59.6%). In patients with acute cerebral infarction, 13 (23.2%) had a poor outcome, and 43 (76.8%) had a good outcome. The fibrinogen level, the proportion of patients with FVHs and the FVHs score in the cerebral infarction group were significantly higher than those in the non-cerebral infarction group (all P<0.05). Multivariate logistic regression analysis showed that the FVHs score was significantly, independently, and positively correlated with the risk of acute cerebral infarction (odds ratio 2.524, 95% confidence interval 1.400-4.552; P=0.002). The National Institutes of Health Stroke Scale score at admission, the modified Rankin Scale score at admission, and FVHs score in the poor outcome group were significantly higher than those in the good outcome group (all P<0.05). Multivariate logistic regression analysis showed that there was a significant independent negative correlation between the FVHs score and the outcome of patients with cerebral infarction (odds ratio 0.144, 95% confidence interval 0.045-0.459; P=0.001). Conclusions:FVHs suggest that the blood supply is in a state of decompensation. When FVHs are present, the risk of cerebral infarction in patients with chronic middle cerebral artery occlusion is significantly increased; the wider the range of FVHs, the higher the risk of poor outcome after cerebral infarction.

Objective: The purpose of this study was to investigate the state of depression and anxiety and assciated factors of back to school college students during the outbreak of COVID-19, so as to provide theoretical basis for emotional counseling and psychological crisis intervention after long term school closure due to epidemic outbreak. Methods: Cluster sampling was used to select the first batch of 2 697 back to school students in a high vocational college. The survey content includes basic information questionnaire, Depression Self rating Scale (SDS) and Anxiety Self rating Scale(SAS). Results: The detection rate of depression and anxiety was 31.8% and 10.4% respectively. Multivariate Logistic regression analysis showed that the risk of depression in female college students was 1.28 times higher than that in male students. The risk of depression among college students with family loss experience caused by the outbreak of COVID-19 was 1.30 times that of those without family loss experience caused by the outbreak of COVID-19. Outgoing personality(OR=0.62), neutral (OR=0.55) and mild temper(OR=0.46), moderate frequency of physical exercise per week(1-2 times:OR=0.73, 3-4 times:OR=0.65) were protective effects for depression in college students.Low frequency of breakfast were the risk effects for depression in college. The risk of anxiety among college students who was the only child in the family was 1.61 times higher than that who was not the only child in the family. The risk of anxiety among college students with family loss experience caused by the outbreak of COVID-19 was 1.45 times higher than that college students without family loss experience caused by the outbreak of COVID-19. Neutral temper, mild temper and physical exercise 1-2 times a week were associated with lower level of anxiety in college students(OR=0.67,0.56,0.67). Breakfast skipping (OR=2.03) was associated with higher levvel of anxiety in college students. Conclusion During the outbreak of COVID-19, the first batch of back to school college students presented high level of depression. Colleges are suggested to promptly carry out effective emotional relief and psychological intervention for students, especially those whose family suffered in COVID-19, the only child in the family, with poor breakfast eating habits, as well as breakfast skipping and lack of physical exercise.

Objective:To investigate the synergistic effect of sarcopenia and osteoporosis on the occurrence of spinal osteoporotic fracture (OPF) in patients with rheumatoid arthritis (RA).Methods:A total of 389 hospitalized RA patients and 156 age and sex-matched normal subjects (control group) were recruited. Dual energy X-ray absorptiometry (DEXA) method was used to measure bone mineral density (BMD) of lumbar spine and hip, and bioelectrical impedance method was applied to determine skeletal muscle mass of limbs. X-ray examination of spin was conducted and spinal OPF was diagnosed according to semi-quality method. Student's t test was used for comparison of measurement date between the two groups, χ2 test was used for comparison of intergroup rates, and Logistic Regression(Backward LR) method was used for multivariate Regression analysis of binomial classification data. Results:BMD of all test sites in RA patients was significantly lower than that in the control group ( P<0.01). The incidence of total OP in RA group was significantly higher than that in the control group [(32.9% vs 12.8%), χ2=22.706, P<0.01]. A total of 84 patients with RA developed spinal OPF, with an incidence of 21.6% which was higher than that in the control group [(3.8%), χ2=25.439, P<0.01]. The incidence of sarcopenia in RA was 54.8%, significantly higher than that in the control group [(9.6%), χ2=93.241, P<0.01]. The incidence of sarcopenia combined with osteoporosis in RA group (28.5%) was significantly higher than that in the control group [(5.8%), χ2=118.110, P<0.01]. Comparison of the incidence of spinal OPF in RA patients among groups with different bone mass (normal bone mass, osteopenia, osteoporosis) showed that the incidence of spinal OPF among these groups was statistically different ( χ2=43.373, P<0.01), and the incidence of spinal OPF increased along with the decrease of bone mass ( χ2=43.003, P<0.01). The incidence of spinal OPF in RA patients with sarcopenia (27.2%, 58/213) was significantly higher than that in RA patients without sarcopenia [(14.8%, 26/176), χ2=8.833, P=0.003]. All participants were divided into three groups: group 1=no OP and sarcopenia, group 2=with sarcopenia or OP, group 3=both sarcopenia and OP. Difference of incidence of spine OPF in RA patients among three groups was statistically significant ( χ2=33.832, P<0.01), and the incidence of spinal OPF raised gradually in group 1 and 3, ( χ2=37.164, P<0.01). Incidences of sarcopenia, OP and spinal OPF in RA treated with glucocorticoid (GC) were higher than those in RA without GC ( P<0.05, P<0.01). Results of logistic regression showed advanced age[ OR(95% CI)=1.069(1.038, 1.101), P<0.01], usage of GC [ OR(95% CI)=3.169(1.679, 5.984), P<0.01] and sarcopenia combined with OP [ OR(95% CI)=2.113(1.430, 3.124), P<0.01] were risk factors for spinal OPF in RA patients. Conclusion:Incidences of sarcopenia, OP and spinal OPF in RA patients are higher than that in normal controls. Sarcopenia and OP have a synergistic effect on spinal OPF in RA patients.