BACKGROUND:Gradient artificial bone repair scaffolds can mimic unique anatomical features in musculoskeletal tissues,showing great potential for repairing injured musculoskeletal tissues. OBJECTIVE:To review the latest research advances in gradient artificial bone repair scaffolds for tissue engineering in the musculoskeletal system and describe their advantages and fabrication strategies. METHODS:The first author of the article searched the Web of Science and PubMed databases for articles published from 2000 to 2023 with search terms"gradient,bone regeneration,scaffold".Finally,76 papers were analyzed and summarized after the screening. RESULTS AND CONCLUSION:(1)As an important means of efficient and high-quality repair of skeletal system tissues,gradient artificial bone repair scaffolds are currently designed bionically for the natural gradient characteristics of bone tissue,bone-cartilage,and tendon-bone tissue.These scaffolds can mimic the extracellular matrix of native tissues to a certain extent in terms of structure and composition,thus promoting cell adhesion,migration,proliferation,differentiation,and regenerative recovery of damaged tissues to their native state.(2)Advanced manufacturing technology provides more possibilities for gradient artificial bone repair scaffold preparation:Gradient electrospun fiber scaffolds constructed by spatially differentiated fiber arrangement and loading of biologically active substances have been developed;gradient 3D printed scaffolds fabricated by layered stacking,graded porosity,and bio-3D printing technology;gradient hydrogel scaffolds fabricated by in-situ layered injections,simple layer-by-layer stacking,and freeze-drying method;and in addition,there are also scaffolds made by other modalities or multi-method coupling.These scaffolds have demonstrated good biocompatibility in vitro experiments,were able to accelerate tissue regeneration in small animal tests,and were observed to have significantly improved histological structure.(3)The currently developed gradient artificial bone repair scaffolds have problems such as mismatch of gradient scales,unclear material-tissue interactions,and side effects caused by degradation products,which need to be further optimized by combining the strengths of related disciplines and clinical needs in the future.

Objective: To systematically review the development and changes in mental health policies within the National Outline for Children s Development in China from 1992 to 2030, providing a reference basis for future formulation of mental health policies among children and adolescent in China. Methods: Based on the four editions of the National Outline for Children s Development in China across different periods from 1992 to 2030, word frequency analysis was used to reveal shifts in policy priorities, and an internationally recognized framework for adolescent health policy analysis was applied to conduct a textual review. Results: Word frequency analysis revealed that the term "psychological" appeared 6 times in the National Outline for Children s Development in China (2001-2010) but increased to 20 times in the National Outline for Children s Development in China (2021-2030) (abbreviated as the National Outline of 2021), while the term "health" rose from 4 times in the National Outline for Children s Development Plan in China in the 1990s to 68 times in the National Outline of 2021. The scope of mental health policy interventions expanded to encompass five key areas:health, safety, education, welfare and legal protection. Textual analysis highlighted that the policies of the National Outline for Children s Development in China were demand driven, prioritized vulnerable groups and continuously broadened their coverage, emphasizing sustainability and appropriateness, and monitoring/evaluation mechanisms. By 2023, 42.3% of primary schools and 64.8% of secondary schools employed full time mental health education teachers. However, the National Outline for Children s Development in China lacked direct evidence of children and adolescents participation in policy formulation, and publicly available mental health data disaggregated by age and gender remained limited. Conclusion Mental health policies of children and adolescents in China have evolved from nonexistence to gradual refinement, yet institutionalized channels for youth involvement in policy development and evaluation remain insufficient, and transparency in age and gender specific mental health data needs improvement.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.

Objective To investigate the expression and clinical significance of CD155 in oral squamous cell carci-noma(OSCC)tissues,as well as its impact on migration,proliferation and invasion.Methods Immunohisto-chemistry(IHC)was used to analyze the expression of CD155 in OSCC tissues and its correlation with clinicopath-ological features and prognosis.The transcription levels of CD155 were assessed by Western blot and RT-qPCR.Cell experiments were conducted to evaluate the effects of CD155 on OSCC cells following transfection with CD155 siRNA.Results CD155 was predominantly expressed on the cell membrane in OSCC tissues,with higher expres-sion levels compared to normal tissues(x2=50.750,P＜0.000 1).High CD155 expression was associated withⅢ+Ⅳ disease stages(x2=25.488,P=0.001),poor differentiation(x2=6.299,P=0.012),T3+T4 depth of invasion(x2=23.820,P=0.001)and lymph node metastasis(x2=7.830,P=0.005)in OSCC patients;survival analysis revealed a correlation between high CD155 expression and shorter patient survival(P＜0.05).COX regression analysis identified lymph node metastasis as an independent factor affecting the survival of OSCC patients(P＜0.05).Both CD155 protein expression and mRNA transcription levels were significantly upregulated in OSCC cells(P＜0.05).Transfection with siRNA-CD155 in OSCC cells resulted in significantly reduced prolif-eration,migration and invasion abilities compared to the control group(P＜0.05).Conclusion CD 155 is highly expressed in OSCC tissues and is associated with poor patient prognosis.Modulating CD155 expression can influ-ence the biological functions of OSCC cells,leading to and inhibition of proliferation,migration,and invasion.

Hiatt-Neu-Cooper neurodevelopmental syndrome(HINCONS),a rare neurodevelopmental disorder closely associated with a heterozygous mutation in the RALA gene on 7p14,is characterized by prominent in-tellectual disability or global developmental delay.Currently,no effective treatment options are available for this dis-ease.This article reports a case of HINCONS presenting with global developmental delay,particularly in language and motor development,along with decreased muscle tone,short stature,distinctive facial features,and congenital ventricular septal defect.No significant abnormalities were found in laboratory biochemical tests.Head MRI revealed enlarged ventricles and increased space in the subarachnoid and convexity regions.Trio-based whole exome sequen-cing testing identified a novel variant RALA gene c.475(exon4)A＞G(p.K159E)in the affected child.In this study,we performed a retrospective analysis of the clinical features of 12 reported cases of HINCONS from the litera-ture,with the aim of providing a reference for the clinical diagnosis and genetic counseling of HINCONS.

Objective:To investigate the protective effect of racanisodamine (654-2) on lung epithelial cell injury induced by X-ray in mice and unravel the underlying mechanism.Methods:Mouse alveolar epithelial cells MLE-12 were used to establish radiation-induced lung injury (RILI) model in vitro and divided into 4 groups as follows: control (no irradiation), radiation (16 Gy radiation), treatment 1 (16 Gy radiation + 2 μmol/L 654-2), treatment 2 (16 Gy radiation + 10 μmol/L 654-2), and inhibitor (16 Gy radiation + 10 μmol/L 654-2 + 2 μmol/L ML385), respectively. Cells were sampled at different time points after radiation. Cell senescence was detected with senescence-associated β-galactosidase (SA-β-Gal) staining. Cell colony-forming ability was detected to observe the recovery capability of cells after treatment. The expression levels of p21, p16, phosphorylated histone H2AX(γH2AX), nuclear factor erythroid-2 related factor 2 (Nrf2), Nrf2 Ser40 site phosphorylation (p-Nrf2), p62, heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1) proteins were measured by Western blot. Cell apoptosis and the production of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The expression levels of glutathione (GSH) and superoxide dismutase (SOD) were detected according to the manufectuer instructions. The expression levels of glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate-cysteine ligase modifier subunit (GCLM) mRNA were determined by real time reverse transcription PCR. Measurement data were expressed as Mean ±SD. Comparison between two groups was conducted by independent sample t-test, and comparison among multiple groups was performed by one-way ANOVA. Results:Compared with the radiation group, the proportion of cells with positive staining of SA-β-Gal was significantly lower and cell senescence were alleviated in the treatment 1 and 2 groups (all P<0.001). Compared with the radiation group, the expression level of γH2AX protein was significantly down-regulated ( P=0.037), cell apoptosis rate was significantly decreased ( P=0.026), the proliferation capacity of MLE-12 was enhanced ( P=0.004), GSH ( P=0.002) and SOD ( P<0.001) activity was enhanced and ROS production was declined ( P=0.001) in the treatment 2 group. The expression levels of Nrf2 and p-Nrf2 in total protein were up-regulated over the time of 654-2 intervention. Meanwhile, the expression levels of antioxidant proteins of NQO1 and HO-1 were up-regulated and that of GCLC and GCLM mRNA was also up-regulated. There were no significant differences in the number of cells with positive staining of SA-β-Gal ( P=0.145) and ROS production ( P=0.317) between the inhibitor and radiation groups after supplement of ML385, small-molecule inhibitor of Nrf2. Conclusion:654-2 can activate the Nrf2 pathway, enhance cell antioxidant capacity and inhibit cell senescence, thereby playing a protective role on radiation- induced lung injury.

Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective:To discuss the expressions of procollagen-lysine,2-oxoglutarate 5-dioxygenase 1(PLOD1)in oral squamous cell carcinoma(OSCC)tissue and OSCC cells and its effect on the biological behavior of the OSCC cells,and to clarify the potential of PLOD1 as a prognostic biomarker for OSCC.Methods:The expression level of PLOD1 mRNA in head and neck squamous cell carcinoma(HNSC)tissue and its correlation with the survival of the HNSC patients were analyzed by Tumor Immune Estimation Resource(TIMER),Gene Expression Profiling Interactive Analysis(GEPIA),and Kaplan-Meier Plotter databases.Immunohistochemistry method was used to detect the expression level of PLOD1 protein in 110 OSCC tissue and 64 adjacent tissue,and its association with clinicopathological characteristics and prognosis of the OSCC patients were analyzed;the diagnostic value of PLOD1 in OSCC was detected by area under the curve(AUC)of the receiver operating characteristic(ROC).The expression levels of PLOD1 mRNA and protein in the human normal oral epithelial HOK cells and OSCC SCC15 and CAL27 cells were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The small interfering RNA(siRNA)fragments were transfected into the SCC15 cells by liposome method and the cells were dividing into si-NC group(transfected with negative control si-NC)and si-PLOD1 group(transfected with si-PLOD1);the proliferation activities,scratch healing rates,and numbers of invasion cells in two groups were detected by CCK-8 method,wound healing assay,and Transwell chamber assay,respectively.Results:The public database analysis results showed that the expression level of PLOD1 mRNA in HNSC tissue was higher than that in adjacent tissue(P＜0.05);compared with low expression of PLOD1 group,the survival period of the HNSC patients in high expression of PLOD1 group was shorter(HR=1.41,P=0.018).The PLOD1 protein was mainly expressed in cytoplasm of the OSCC cells,and the expression intensity of PLOD1 in OSCC tissue was higher than that in adjacent tissue(P＜0.01),and it was related to T stage and TNM stage of the OSCC patients(P=0.021,P=0.004).The AUC of PLOD1 for diagnosing OSCC was 0.811,and the specificity was 63.64％and the sensitivity was 90.63％.The Kaplan-Meier survival analysis results showed that compared with low expression of PLOD1 group,the survival rate of the OSCC patients in high expression of PLOD1 group was decreased(P＜0.01).The COX regression analysis results showed that high expression of PLOD1 was an independent risk factor for the prognosis of OSCC(P=0.012).The expression levels of PLOD1 mRNA and protein in the OSCC cells were higher than those in HOK cells(P＜0.05).Compared with si-NC group,the proliferation activity and scratch healing rate of the cells in si-PLOD1 group were decreased(P＜0.05),and the number of invasion cells was decreased(P＜0.01).Conclusion:PLOD1 is highly expressed in the OSCC tissue and cells,and silencing of PLOD1 expression can inhibit the proliferation,migration,and invasion of the OSCC cells.PLOD1 may be a new therapeutic target and prognostic biomarker for OSCC.

Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective To investigate the status quo and its influencing factors of nurses'organizational silence in 3 Grade A general hospitals.Methods Convenient sampling method was used to investigate clinical nurses in 3 Grade A general hospitals in Xi'an from April to August 2023 by general data questionnaire,nurses'organizational silence questionnaire and hospital magnetic factor scale.Multiple linear regression was used to analyze the influencing factors of organizational silence.Results A total of 855 nurses completed the study.The total silence score of nurses was(56.33±8.55);The total score of hospital magnetic level was(107.63±12.85).There was a negative correlation between nurse tissue silence and hospital magnetic level(r=-0.318,P<0.01).Hospital magnetic level,age,job title and working time were the influential factors of nurses'organizational silence(all P<0.001),which together explained 62.60%of the variation.Conclusions The silence of nurses'tissue and the level of hospital magnetism are in the low-medium level.Nurses are younger in age,lower in professional title,shorter in nursing age and lower in hospital magnetism level,the higher the tissue age level is,the nursing managers can reduce the tissue silence of nurses by improving the hospital magnetism level.