BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Objective:To establish the reference values and growth curves of skeletal muscle mass among children in the Nanjing area.Methods:A cross-sectional study was conducted with children who underwent physical examination at the Department of Child Health Care, Children′s Hospital of Nanjing Medical University from 2020 January to 2022 September. Their height, weight, body fat mass and skeletal muscle mass were measured. Body mass index, percentage of body fat mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat were calculated. The associations between skeletal muscle mass indices and physical measurements index were analyzed through the Spearman correlation test. The Mann-Kendall test was used to assess the trend for skeletal muscle mass. Generalized additive models for location, scale and shape were used to construct percentile reference values and growth curves of male and female skeletal muscle mass indices at different ages.Results:A total of 32 690 children aged 4-14 years were enrolled in this study, including 19 912 boys (60.91%). Skeletal muscle mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat of boys and girls was 11.10 (8.40, 14.90) and 10.30 (7.90, 13.20) kg, 40.36% (37.01%, 43.13%) and 39.38% (36.43%, 41.88%), 6.70 (6.07, 7.52) and 6.33 (5.79, 7.00), 2.39 (1.46, 3.47) and 2.14 (1.45, 3.00) kg/m 2, respectively. Skeletal muscle mass of both boys and girls was all positively associated with weight ( r=0.97, 0.96), body mass index ( r=0.68, 0.63) and percentage of body fat mass ( r=0.40, 0.43) (all P<0.01). The reference values and growth curves showed that the percentage of skeletal muscle mass P50 ranged from 37.75%-44.61% in boys and from 36.22%-40.55% in girls. The relative skeletal muscle mass index P50 ranged from 5.80-9.68 kg/m 2 in boys and from 5.57-7.98 kg/m 2 in girls. The ratio of skeletal muscle to body fat P50 ranged from 1.86-2.67 in boys and from 1.29-2.41 in girls. There was an increasing trend with age for both boys and girls in the growth of skeletal muscle mass ( Z=4.20, 3.75, both Ptrend<0.01), and increased slightly before 9 years of age and then increased rapidly until 14 years of age in both boys and girls. Conclusions:The skeletal muscle mass indices change with age and gender during childhood. Percentile reference values for pediatric skeletal muscle mass indices can be used to evaluate the muscular growth and development in children in the Nanjing area.

Objective:To investigate the predictive value of neck circumference on cardiometabolic risk in children.Methods:This was a cross-sectional study of natural sources. As the prediction cohort, clinical data were collected from 3 443 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from July 2021 to September 2022. As the validation cohort for external validation, clinical data were collected from 604 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from October 2022 to March 2023. Height, weight, neck circumference, waist circumference and body composition were measured in both groups, and body mass index, neck circumference to height ratio (NHtR), waist circumference to height ratio, body fat percentage and skeletal muscle percentage were calculated. Systolic blood pressure, diastolic blood pressure, fasting blood glucose, blood lipid and uric acid and other cardiovascular and metabolic risk indicators were collected in both groups. The prediction cohort was further stratified into clustered and non-clustered groups based on the clustering of cardiometabolic risk factors (CCRF). Various variables between these 2 groups were compared using the Mann-Whitney U test. Pearson correlation and binary Logistic regression were conducted to investigate the correlations between neck circumference and cardiovascular metabolic risk factors. The accuracy of NHtR in predicting the CCRF was evaluated using the area under the curve (AUC) of receiver operating characteristic (ROC). The cutoff value was determined using the Youden index. The validation cohort was then divided into groups above and below the cutoff value, and the detection rate of CCRF between the 2 groups was compared using the χ2 test for validation .Results:In the prediction cohort of 3 443 children (2 316 boys and 1 127 girls), 1 395 (40.5%) children were overweight or obese, and 1 157 (33.6%) children had CCRF. Pearson correlation analysis revealed all significant positive correlations (all P<0.01) between neck circumference and systolic blood pressure ( r=0.47, 0.39), diastolic blood pressure ( r=0.27, 0.21), uric acid ( r=0.36, 0.30), and triglycerides ( r=0.20, 0.20) after adjusting for age in both males and females. Among both males and females, neck circumference both showed significant negative correlation (both P<0.01) with high-density lipoprotein cholesterol ( r=-0.27, -0.28), and no correlation with fasting glucose levels ( r=0.03, -0.03, both P>0.05). After adjusting for gender, age, and body fat percentage, increased body mass index, neck circumference, or waist circumference increased the risks of hypertension ( OR=1.23, 1.39, 1.07, all P<0.001), hyperuricemia ( OR=1.16, 1.23, 1.05, all P<0.001), hypertriglyceridemia ( OR=1.08, 1.16, 1.02, all P<0.01), low high-density lipoprotein cholesterol ( OR=1.10, 1.27, 1.03, all P<0.01), and the CCRF ( OR=1.51, 1.73, 1.15, all P<0.01). The areas under the ROC curves of NHtR in predicting CCRF was 0.73, with sensitivity and specificity at 0.66 and 0.71, respectively. The corresponding optimal cut-off value was 0.21. Validation with 604 children confirmed that the detection of CCRF in the NHtR≥0.21 group was 3.29 times (60.5% (112/185) vs. 18.7% (79/422), χ2=107.82, P<0.01) higher compared to the NHtR <0.21 group. Conclusions:Neck circumference is associated with cardiovascular metabolic risks such as hypertension, hyperlipidemia, hyperglycemia, and hyperuricemia in children. When the NHtR is ≥0.21, there is an increased likelihood of CCRF.

Objective:To investigate the value of intratumoral and peritumoral radiomics models based on 18F-FDG PET-CT in predicting epidermal growth factor receptor (EGFR) mutation status in lung adenocarcinoma and interpret peritumoral radiomics features. Methods:This study was a cross-sectional study. Patients with lung adenocarcinoma who underwent 18F-FDG PET-CT at the Third Affiliated Hospital of Soochow University between January 2018 and April 2022 were retrospectively collected and samplied into a training set (309 cases) and a test set (206 cases) in a 6∶4 ratio randomly. Radiomics features were extracted from the intratumoral and peritumoral regions of interest based on PET and CT images, respectively, and the optimal feature sets were selected. Radiomics models were established using the XGBoost algorithm, and radiomics scores (intratumoral CT label, peritumoral CT label, intratumoral PET label, peritumoral PET label) were calculated. Logistic regression analysis was used to construct a clinical model and a combined model (incorporating PET-CT intratumoral and peritumoral radiomics, clinical features, and CT semantic features). The predictive performance of the models was evaluated using receiver operating characteristic curves and the area under the curve (AUC). Unsupervised clustering, Spearman correlation analysis, and visualization methods were used for the interpretability of peritumoral radiomics features. Results:In both the training and test sets, the AUC value of CT peritumoral labels was greater than that of CT intratumoral labels for predicting EGFR mutation status in lung adenocarcinoma (training set: Z=3.84, P<0.001; test set: Z=1.99, P=0.046). In the test set, the AUC value of PET intratumoral labels (0.684) was slightly higher than that of PET peritumoral labels (0.672) for predicting EGFR mutation status, but the difference was not statistically significant ( P>0.05). The combined model had the highest AUC value for predicting EGFR mutation status of lung adenocarcinoma in both the training and test sets and was significantly better than the clinical model (training set: Z=6.52, P<0.001; test set: Z=2.31, P=0.021). Interpretability analysis revealed that CT peritumoral radiomics features were correlated with CT shape features, and there were significant differences in CT peritumoral features between different EGFR mutation statuses. Conclusions:The value of CT peritumoral labels is superior to that of CT intratumoral labels in predicting EGFR mutation status in lung adenocarcinoma. The predictive performance of the model can be improved by combining PET-CT intratumoral and peritumoral radiomics, clinical features, and CT semantic features.

Nasopharyngeal carcinoma (NPC) is the third most common malignancy with a high recurrence and metastasis rate in South China. Natural compounds extracted from traditional Chinese herbal medicines have been developed and utilized for the treatment of a variety of cancers with modest properties and slight side effects. Maackiain (MA) is a type of flavonoid that was first isolated from leguminous plants, and it has been reported to relieve various nervous system disorders and exert anti-allergic as well as anti-inflammatory effects. In this study, we demonstrated that MA inhibited proliferation, arrested cell cycle and induced apoptosis in nasopharyngeal carcinoma CNE1 and CNE2 cells in vitro and in vivo. The expression of the related proteins associated with these processes were consistent with the above effects. Moreover, transcriptome sequencing and subsequent Western blot experiments revealed that inhibition of the MAPK/Ras pathway may be responsible to the anti-tumor effect of MA on NPC cells. Therefore, the effects of MA and an activator of this pathway, tertiary butylhydroquinone (TBHQ), alone or combination, were investigated. The results showed TBHQ neutralized the inhibitory effects of MA. These data suggest that MA exerts its anti-tumor effect by inhibiting the MAPK/Ras signaling pathway and it has the potential to become a treatment for patients with NPC.
Humans ; Nasopharyngeal Carcinoma/pathology* ; Cell Line, Tumor ; Cell Proliferation ; Apoptosis ; Signal Transduction ; Nasopharyngeal Neoplasms/pathology*

Obesity is the main risk factor of cardiovascular diseases.As the indexes of obesity evaluation, body mass index(BMI) and waist circumference are the most common methods to predict metabolic risk of cardiovascular system.Recently, neck circumference, a reliable and low-cost parameter of anthropometry, could be considered as a new indicator to evaluate childhood obesity since it is closed correlation with children′s BMI and waist circumference.Moreover, studies had shown that the increase of neck circumference had strong correlation with cardiometabolic risk, such as high blood pressure, hyperlipidemia, hyperglycaemia and hyperuricemia.This indicates that it might be the index to predict metabolic risk of cardiovascular system.Understanding and illuminating the changes of neck circumference and its relationship with obesity and metabolic disorders would have significances for preventing and identifying metabolic risks of cardiovascular system in children and adolescents.

Objective To investigate the possible action mechanism of the micro ribonucleic acid-1-3p(miR-1-3p)/Annexin A2(AnxA2)molecular axis in high glucose(HG)-induced neovascularization of human retinal microvascular en-dothelial cells(HRMECs).Methods A cell injury model was established by culturing HRMECs in vitro and treating them with HG.The HRMECs were divided into the Con group(DMEM medium containing fetal bovine serum in volume fraction of 10％),HG group(cultured in 25 mmol·L-1 D-glucose),HG+miR-NC group(transfected with miR-NC),HG+miR-1-3p group(transfected with miR-1-3p mimics),HG+sh-NC group(transfected with sh-NC),HG+sh-AnxA2 group(transfect-ed with sh-AnxA2),HG+miR-1-3p+pcDNA group(transfected with miR-1-3p mimics+pcDNA),and HG+miR-1-3p+pcDNA-AnxA2 group(transfected with miR-1-3p mimics+pcDNA-AnxA2).After 48 h of transfection,cells were collected and cultured in 25 mmol·L-1 D-glucose medium for 24 h.Cell viability and number of migrating cells were detected using MTT and Transwell chamber experiments,respectively.The number of lumen formations was detected by the lumen forma-tion experiment.The dual luciferase reporter assay was adopted to detect the targeting relationship between miR-1-3p and AnxA2.Western blot was used to detect the protein levels of vascular endothelial growth factor(VEGF)and matrix metal-loproteinase-2(MMP-2).Results Compared with the Con group,the expression level of miR-1-3p in the HG group de-creased,while the levels of AnxA2 messenger ribonucleic acid(mRNA)and protein increased,with statistically significant differences(all P＜0.05).Compared with the Con group,the HG group showed an increase in cell viability,number of mi-grating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically significant differences(all P＜0.05).Compared with the HG+miR-NC group,the HG+miR-1-3p group showed a decrease in cell viability,number of migrating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically significant differences(all P＜0.05).Compared with the HG+sh-NC group,the HG+sh-AnxA2 group showed a decrease in cell viability,number of mi-grating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically significant differences(all P＜0.05).Compared with the HG+miR-1-3p+pcDNA group,the HG+miR-1-3p+pcDNA-AnxA2 group showed an increase in cell viability,number of migrating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically signifi-cant differences(all P＜0.05).Conclusion Overexpression of miR-1-3p can inhibit proliferation,migration and neovas-cularization of HRMECs by targetedly regulating AnxA2 expression.

Objective:To investigate the correlation between the SUV index (SUV max of the lesion/SUV mean of the liver) in 18F-FDG PET/CT imaging and the invasiveness of early lung adenocarcinoma presenting as ground-glass nodule (GGN). Methods:From January 2012 to March 2020, 167 GGN patients (49 males, 118 females; age: (61.5±9.0) years) with early lung adenocarcinoma who underwent PET/CT imaging in Changzhou First People′s Hospital were retrospectively enrolled. The image parameters including the GGN number, location, type, edge, shape, abnormal bronchus sign, vacuole sign, pleural depression, vessel convergence sign, GGN diameter ( DGGN), solid component diameter ( Dsolid), consolidation to tumor ratio (CTR, Dsolid/ DGGN), CT values (CT value of ground-glass opacity (CT GGO), CT value of lung parenchyma (CT LP), ΔCT GGO-LP (CT GGO-CT LP)) and SUV index were analyzed. Single and multivariate logistic regressions were used to analyze the correlation between SUV index and infiltration. The generalized additive model was used for curve fitting, and the piece-wise regression model was used to further explain the nonlinearity. Results:In 189 GGNs, invasive adenocarcinoma accounted for 85.2% (161/189). Single logistic regression showed that the GGN number, type, shape, edge, abnormal bronchus sign, pleural depression, vessel convergence sign, DGGN, Dsolid, CTR, CT GGO, ΔCT GGO-LP and SUV index were related factors of infiltration (odds ratio ( OR) values: 0.396-224.083, P<0.001 or P<0.05). After fully adjusting for confounding factors, SUV index was significantly correlated with increased risk of invasion ( OR=2.162 (95% CI: 1.191-3.923), P=0.011). Curve fitting showed that the SUV index was non-linearly related to the risk of infiltration, and the risk of infiltration increased significantly only when the SUV index was greater than 0.43 ( OR=3.509 (95% CI: 1.429-8.620), P=0.006). The correlation between SUV index and infiltration had no interaction between age, vacuoles, pleural depression and CTR subgroups (all P>0.05). Conclusions:SUV index is an independent factor related to the invasiveness of early lung adenocarcinoma. The higher the SUV index, the greater the risk of invasion; but the two are not simply linearly correlated.

Objective:To analyze the postoperative paravertebral muscle degeneration and its correlations with health related quality of life (HRQL) in patients undergoing minimally invasive surgery-transforaminal lumbar interbody fusion (MIS-TLIF).Methods:The clinical data of the 50 patients were retrospectively analyzed who had undergone single-segmental MIS-TLIF at Department of Spinal Cord Surgery, The People's Hospital of Henan Province from January 2019 to December 2021. The relative volumes of lumbar posterior muscle (LM), the relative volumes of the psoas major (PM), and the rates of fatty degeneration (FD) of the fused segment and its adjacent segments were compared respectively between preoperation, 6 and 12 months postoperation. The correlations were analyzed between the HRQL scores [visual analog scale (VAS) for pain and Oswestry disability index (ODI)] and the relative LM volumes, the relative PM volumes, and the FD rates of the fused segment and its adjacent segments at 12 months postoperation.Results:Compared with the preoperative values, the relative LM volumes and the relative PM volumes of the fused segment and its adjacent segments at 6 and 12 months postoperation were significantly reduced while the FD rates significantly increased. However, the FD rate of the fused segment at 12 months postoperation (20.6% ± 6.1%) was significantly lower than that at 6 months postoperation (29.7% ± 8.2%) ( P < 0.05). The VAS score was strongly negatively or positively correlated with the relative LM volume ( r = -0.819, P < 0.001) and the FD rate ( r = 0.86, P < 0.001) of the fused segment, and moderately negatively correlated with the relative PM volume ( r = -0.435, P = 0.016). The ODI index was moderately negatively correlated with the relative LM volume ( r = -0.512, P = 0.004) and the relative PM volume ( r = -0.402, P = 0.020) of the fused segment, but moderately positively correlated with the FD rate of the fused segment ( r = 0.565, P = 0.001). There was a moderate negative correlation between the ODI index and the relative LM volume of the adjacent segments ( r = -0.478, P = 0.012). Conclusions:After MIS-TLIF, the volume of the paravertebral muscles decreases and the dorsal muscles develop fatty degeneration. The improvement of LM fatty degeneration may be observed by 12-month follow-up in the fused segment, but not in the adjacent segments. The LM volume and the FD rate of the fused segment are the most closely related to the postoperative HRQL.