The IFT80/Hedgehog Pathway Regulates the Osteogenic-adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells.

BACKGROUND: Steroid-induced avascular necrosis of the femoral head (SANFH) is a typical refractory disease that often progresses irreversibly and has a high disability rate. Numerous studies have confirmed that abnormal osteogenic-adipogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) is one of the major factors of SANFH. However, the mechanism remains to be elucidated. OBJECTIVES: This study aimed to investigate the mechanism and effect of the IFT80/Hedgehog-mediated osteogenic-adipogenic differentiation of BM-MSCs in SANFH. METHODS: Femoral head specimens of SANFH patients and femoral neck fractures (FNF) patients were collected to detect the expression of IFT80, Shh and osteogenic-adipogenic differentiation-related genes by immunohistochemistry (IHC), western blot (WB) and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR). Based on the rabbit SANFH model, the mRNA expression and protein level of IFT80 and Shh were detected by RT-qPCR and WB. After the osteogenic/adipogenic differentiation based on rabbit BM-MSCs, the IFT80, Gli1, PPAR-γ, and Runx2 expression were detected. Differences in alkaline phosphodiesterase activity, calcium nodule, quantification/distribution of lipid droplets, expression of IFT80/Hedgehog axis, and the level of osteogenic- adipogenic associated factors were determined after IFT80 overexpression. RESULTS: RT-qPCR, WB and IHC revealed that IFT80 and Shh lowly expressed in the osteoblasts and intra-trabecular osteocytes at the edge of trabeculae and in the intercellular matrix of the bone marrow lumen in the SANFH specimens. The Runx2 expression was low, while the PPAR-γ expression was high in both human specimens and animal models of SANFH, suggesting that the balance of osteogenic-adipogenic differentiation was dysregulated. Rabbit BM-MSCs with stable overexpression of IFT80 showed increased alkaline phosphatase activity after induction of osteogenic differentiation, increased calcium nodule production, and decreased adipogenesis after induction of adipogenic differentiation. CONCLUSION: There is a dysregulation of the balance of osteogenic-adipogenic differentiation in SANFH. IFT80 may inhibit adipogenic differentiation while promoting osteogenic differentiation in rabbit BM-MSCs by activating the Hedgehog pathway.

Verification of cuproptosis-related diagnostic model associated with immune infiltration in rheumatoid arthritis.

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease closely related to inflammation. Cuproptosis is a newly discovered unique type of cell death, and it has been found that it may play an essential role in the occurrence and development of RA. Therefore, we intend to explore the potential association between cuproptosis-related genes (CRGs) and RA to provide a new biomarker for the treatment and prognosis of RA. Methods: Download GSE93777 datasets from the GEO database. Variance analysis was performed on the CRGs that had been reported. Then, the random forest (RF) model and nomogram of differentially expressed CRGs were constructed, and the ROC curve was used to evaluate the accuracy of the diagnostic model. Next, RA patients were subtyped by consensus clustering, and immune infiltration was analyzed in each subgroup to confirm the correlation between CRGs and abundance of immune cells. The expression levels of CRGs were verified by qRT-PCR. Results: Eight differentially expressed CRGs (DLST, DLD, PDHB, PDHA1, ATP7A, CDKN2A, LIAS, DLAT) were screened out by differential analysis to construct an RF model. The ROC curve proved that this model had good diagnostic accuracy. Based on the above eight significant CRGs, a nomogram was built to predict effective and high-precision results. The consensus clustering method identified two CRG patterns. Most of the immune cells were enriched in cluster A, indicating that cluster A may be related to the development of RA. Finally, qRT-PCR verified the expression of eight key genes, further confirming our findings. Conclusion: The diagnosis model of RA based on the above eight CRGs has excellent diagnostic potential. Based on these, patients can be divided into two different molecular subtypes; it is expected to develop a new treatment strategy for RA.

Exosome-mediated miR-144-3p promotes ferroptosis to inhibit osteosarcoma proliferation, migration, and invasion through regulating ZEB1.

BACKGROUND: Osteosarcoma (OS) is the most prevalent orthopedic malignancy with a dismal prognosis. The high iron absorption rate in OS cells of patients suggests that ferroptosis may be related to the progression of OS, but its potential molecular regulatory role is still unclear. Based on the ability to couple with exosomes for targeted delivery of signals, exosome-derived micro ribonucleic acids (miRNAs) can potentially serve as diagnostic biomarkers for OS. METHODS: We identified ferroptosis-related miRNAs and messenger ribonucleic acids(mRNAs) in OS using bioinformatics analysis and performed survival analysis. Then we measured miRNA expression levels through exosome microarray sequencing, and used RT-qPCR and IHC to verify the expression level of miR-144-3p and ZEB1. Stable gene expression cell lines were fabricated for in vitro experiments. Cell viability, migration and invasion were determined by CCK-8 and transwell experiment. Use the corresponding reagent kit to detect GSH/GSSG ratio, Fe2+ level, MDA level and ROS level, and measure the expression levels of GPX4, ACSL4 and xCT through RT-qPCR and WB. We also constructed nude mice model for in vivo experiments. Finally, the stability of the miRNA/mRNA axis was verified through functional rescue experiments. RESULTS: Low expression of miR-144-3p and high expression of ZEB1 in OS cell lines and tissues was observed. Overexpression of miR-144-3p can promote ferroptosis, reduce the survival ability of OS cells, and prevent the progression of OS. In addition, overexpression of miR-144-3p can downregulate the expression of ZEB1 in cell lines and nude mice. Knockdown of miR-144-3p has the opposite effect. The functional rescue experiment validated that miR-144-3p can regulate downstream ZEB1, and participates in the occurrence and development of OS by interfering with redox homeostasis and iron metabolism. CONCLUSIONS: MiR-144-3p can induce the occurrence of ferroptosis by negatively regulating the expression of ZEB1, thereby inhibiting the proliferation, migration, and invasion of OS cells.

Spectral optimization of trichromatic white LEDs based on age of lighting user and application scene.

The optimization of trichromatic white light emitting diodes (LEDs) spectrum for application scenes related to the age of lighting users is proposed and demonstrated. Based on the spectral transmissivity of human eyes at different ages, the visual and non-visual responses of human eyes to different wavelengths of light, we have built the blue light hazards (BLH) and circadian action factor (CAF) related to the age of the lighting user. The BLH and CAF are used to evaluate the spectral combinations of high color rendering index (CRI) white LEDs obtained from different radiation flux ratios of red, green, and blue monochrome spectrum. The best spectra of white LEDs for lighting users at different ages in work and leisure scenes are achieved due to the optimization criterion of BLH proposed by us. This research provides a solution for intelligent health lighting design applicable to light users of different ages and application scenes.

Comprehensive analysis of protein phosphatase 1 regulatory inhibitor subunit 14B, a molecule related to tumorigenesis, poor prognosis, and immune cell infiltration in lung adenocarcinoma.

OBJECTIVE: To explore the relationship between Protein Phosphatase 1 Regulatory Inhibitor Subunit 14B (PPP1R14B) and the occurrence of lung adenocarcinoma (LUAD). METHOD: PPP1R14B expression was investigated using various databases, and its molecular functions and pathways were evaluated using Gene Set Variation Analysis (GSVA) and Gene Set Enrichment Analysis (GSEA). Then, the correlation between tumor mutations and PPP1R14B expression was analyzed. Furthermore, the regulation network and expression pathway axes of PPP1R14B were constructed. The correlation analysis between PPP1R14B and immune cell infiltration was performed using deconvolution algorithm analysis and the Tumor Immune Dysfunction and Exclusion (TIDE) algorithm. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical (IHC) staining of the clinical samples were used for expression validation. RESULTS: PPP1R14B showed high expression in tumor tissue. PPP1R14B was associated with T and N stages and poor prognosis and was linked to the cell cycle, DNA repair, and low immune response. High PPP1R14B expression was associated with high tumor mutation rates. The upstream and downstream genes of PPP1R14B were identified, along with the construction of a protein-protein interaction network (PPI network) and the expression pathway axes of PPP1R14B. PPP1R14B expression was associated with poor immune cell infiltration and a negative correlation between PPP1R14B and mast cell and eosinophil infiltration. CONCLUSION: This study reveals high PPP1R14B expression in LUAD, its contribution to poor prognosis, molecular function, biological pathways, and impact on immune cell infiltration, and provides great insight into the role of PPP1R14B in LUAD tumorigenesis.

Verification of Ferroptosis Subcluster-Associated Genes Related to Osteosarcoma and Exploration of Immune Targeted Therapy.

Background: Despite tremendous advances in treating osteosarcoma (OS), the survival rates of patients have failed to improve dramatically over the past decades. Ferroptosis, a newly discovered iron-dependent type of regulated cell death, is implicated in tumors, and its features in OS remain unascertained. We designed to determine the involvement of ferroptosis subcluster-related modular genes in OS progression and prognosis. Methods: The OS-related datasets retrieved from GEO and TARGET database were clustered for identifying molecular subclusters with different ferroptosis-related genes (FRGs) expression patterns. Weighted gene coexpression network analysis (WGCNA) was applied to identify modular genes from FRG subclusters. The least absolute shrinkage and selection operator (LASSO) algorithm and multivariable Cox regression analysis were adopted to develop the prognostic model. Potential mechanisms of development and prognosis in OS were explored by gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis (GSEA). Then, a comprehensive analysis was conducted for immune checkpoint markers and assessment of predictive power to drug response. The protein expression levels of the three ferroptosis subcluster-related modular genes were verified by immunohistochemistry. Results: Two independent subclusters presenting diverse expression profiles of FRGs were obtained, with significantly different survival states. Ferroptosis subcluster-related modular genes were screened with WGCNA, and the GESA results showed that ferroptosis subcluster-related modular genes could affect the cellular energy metabolism, thus influencing the development and prognosis of osteosarcoma. A prognostic model was established by incorporating three ferroptosis subcluster-related modular genes (LRRC1, ACO2, and CTNNBIP1) and a nomogram by integrating clinical features, and they were evaluated for the predictive power on OS prognosis. The 20 immune checkpoint-related genes confirmed the insensitivity to tumor immunotherapy in high-risk patients. IC50s of Axitinib and Cytarabine suggested a higher sensitivity to the targeted drug. Finally, the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry were consistent with bioinformatics analysis. Conclusion: Ferroptosis are closely associated with the OS prognosis. The risk-scoring model incorporating three ferroptosis subcluster-related modular genes has shown outstanding advantages in predicting patient prognosis.

Health-related quality of life (HRQoL) associated with echinococcosis patients in Tibetan communities in Shiqu County, China: a case-control study.

PURPOSE: Echinococcosis is a chronic neglected zoonotic disease with high endemic in western China. The aim of the study was to evaluate the Health-related Quality of Life (HRQoL) in patients with echinococcosis in Tibetan communities for investigating the physical and mental health challenges faced by the echinococcosis patients and meeting community health needs. METHODS: The HRQoL was measured with 36-Item Short Form Health Survey (SF-36) questionnaire in the case-control study. Analysis of variance (ANOVA) method was used to analyze the data. RESULTS: 50 echinococcosis patients were collected in study group and divided into three subgroups by the duration of staying on albendazole tablets. 52 healthy individuals were collected in control group. In physical quality of life, the duration of staying on albendazole tables influenced the PF (F = 6.661, P < 0.001), RP (F = 3.130, P = 0.029), and GH (F = 4.105, P = 0.008). In mental quality of life, the duration of staying on albendazole tables influenced the SF (F = 10.764, P < 0.001) and RE (F = 5.573, P = 0.001). The income level was the important confounder in PF (F = 11.515, P = 0.001), GH (F = 10.315, P = 0.002), VT (F = 5.984, P = 0.016), and MH (F = 5.565, P = 0.020). CONCLUSIONS: The HRQoL in people with echinococcosis is reduced in comparison with the healthy individuals. It is necessary to shorten the duration of treatment and adopt the special methods adapting the local culture to improve the health-related quality of life in Tibetan communities. Meanwhile, the economic improvement is the foundation.

Genetic polymorphism of the plasminogen activator inhibitor-1 is associated with an increased risk of endometrial cancer.

BACKGROUND AND OBJECTIVES: To investigate the association of uPA system genes, including uPA, uPA receptor (uPAR), and plasminogen activator inhibitor (PAI)-1 gene polymorphisms, with risk of endometrial cancer. METHODS: In the present case control study, we enrolled a total of 134 patients with endometrial cancer confirmed by histopathology and 302 unrelated healthy individuals. Genetic polymorphisms of uPA system genes, including uPA rs4065 C/T SNP, uPAR rs344781 T/C SNP, and PAI-1 rs1799889 4G/5G SNP were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotyping analysis. RESULTS: Frequency of PAI-1 rs1799889 4G/4G genotype and 4G allele differed significantly between patients with endometrial cancer (36.6% and 61.6%, respectively) and healthy individuals (25.5% and 52.2%, respectively). Individuals with PAI-1 rs1799889 4G/4G genotype were at higher risk of endometrial cancer (OR = 2.26; 95% CI: 1.20-4.27). Stratification analysis showed individuals with PAI-1 rs1799889 4G/4G genotype were at elevated risk for endometrioid type (OR = 2.49; 95% CI 1.27-4.88), low stage (stages I-II) endometrial cancer (OR = 2.34; 95% CI 1.21-4.52). However, no significant differences in uPA C/T SNP, uPAR T/C SNP genotypes were observed between endometrial carcinoma cases and controls. CONCLUSIONS: Individuals with PAI-1 rs1799889 4G/4G genotype were at significantly higher risk of endometrial cancer in this study.

Antiobesity and hypolipidemic effects of polyphenol-rich longan (Dimocarpus longans Lour.) flower water extract in hypercaloric-dietary rats.

Plenty of polyphenols, i.e. phenolic acids and flavonoids, were found in longan flower water extract (LFWE) through spectrophotometric and HPLC analyses. Antiobesity and hypolipidemic effects of polyphenol-rich longan flower water extract (LFWE) were investigated in this study. Eight male rats per group were assigned randomly to one of the following dietary groups: (1) normal-caloric diet and pure water (NCD + NDW); (2) hypercaloric diet and pure water (HCD + NDW); (3) HCD and 1.25% (w/v) LFWE (HCD + 1.25% LFWE); (4) HCD and 2.5% (w/v) LFWE (HCD + 2.5% LFWE) for 9 weeks. Body weight, size of epididymal fat, serum triglyceride level and atherogenic index, and hepatic lipids were decreased (p < 0.05) in HCD rats by drinking 2.5% LFWE which may result from downregulated (p < 0.05) pancreatic lipase activity, and sterol regulatory element binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) gene expressions, as well as upregulated (p < 0.05) LDL receptor (LDLR) and peroxisome proliferator-activated-receptor-alpha (PPAR-alpha) gene expressions, and also increased (p < 0.05) fecal triglyceride excretions. Therefore, polyphenol-rich LFWE indeed characterizes antiobesity and hypolipidemic effects in vivo.

Desulphurization of coal via low temperature atmospheric alkaline oxidation.

Different from other options which usually required strict conditions, a method combining atmospheric oxidization and chemical cleaning with alkali solutions was employed to desulphur coals at temperature around 90 degrees C. The data show that 66% organic sulphur, 44% sulphide sulphur, and 15% pyrite sulphur were lost when the coal was treated in 0.25M NaOH at 90 degrees C, while the solution being aerated at the flow rate of 0.136m3h(-1). The rate increased to 73% for organic sulphur, 83% for sulphide sulphur and 84% for pyrite sulphur when the previous coal was further treated in acidic solution containing HCl at pH 1 for another hour. The mechanism of desulphurization was explored using inductively coupled plasma mass spectrometry, X-ray photoelectron spectroscopy and infrared. It was found out that the bond of -CS was broken by atmospheric oxygen in basic environment, leading to the lost of organic sulphur in coal. Scanning electron microscope data show that the physical structure of the coal was not adversely affected by the treatment and thermogravimetric analysis results prove that the pyrolysis behavior remained unchanged, indicating that the burning process of the coal would not be adversely affected. Unlike other oxidizing methods, this technique does not lower the heating value of the coal which was manifested by relevant data.

[The assessment of criminal responsibility of schizophrenic rape offenders].

OBJECTIVE: To explore potential application of Diminished Criminal Responsibility Rating Scale (DCRRS) on rape offenders. METHODS: The DCRRS was used retrospectively to study 77 cases of rape offenders. They were divided into three groups according to the degree of diminished criminal responsibility. RESULTS: There were significant differences in rating scores among three groups (mild group 21.32+/-5.56, moderate group 25.92+/-5.19, and major group 29.54+/-4.62), and 16 of 18 items showed good correlation with the total scores of the scale (r=0.256 to 0.611), 7 factors were extracted by the factor analysis and their cumulative squared loadings was 75.784%. CONCLUSION: The DCRRS is valid and reliable in a in assessment of criminal responsibility schizophrenic rape offenders.

Optimization and thermodynamic assessment of ferrite (Fe3O4) synthesis in simulated wastewater.

The effect of temperature, pH, the rate and time of oxidation, the concentration of ferrous ion in the starting suspensions as well as the amount of oxidant acting on the process of Fe(3)O(4) synthesis by Fe(OH)(2) suspensions are investigated. After 2h reaction at 90 degrees C under the oxidation of 10 g/L NH(4)NO(3), solution containing 0.25-0.35 mol/L iron(II) ion initially would yield the greatest amount of Fe(3)O(4), up to 95% Fe(3)O(4) could be formed. pH of the solution should be controlled between 9.0 and 11.0. X-ray diffraction (XRD) analysis shows that the product has spinel structure, which indicated that the product is Fe(3)O(4). Transmission electron microscopy (TEM) images show that the crystal size of ferrite is around 0.2 microm. The equilibrium composition of the synthesis reaction of Fe(3)O(4) is optimized by the minimization of the free energy of thermodynamics. It was found that the optimal condition for the synthesis of Fe(3)O(4) obtained through experiment is correspondent with that obtained through computer calculation. In the Fe(3)O(4) formation area given by Kiyama [M. Kiyama, Conditions for the formation of Fe(3)O(4) by the air oxidation of Fe(OH)(2) suspensions, Bull. Chem. Soc. Jpn. 47 (7) (1974) 1646-1650], the content of the product formed is not the same everywhere. The main factor that influences the content of the product is the amount of oxidant.

Synthesis of ferrites obtained from heavy metal solutions using wet method.

Wet method was employed to the treatment of heavy metal-contaminated wastewater, and Zn(x)Fe(3-x)O(4), Ni(x)Fe(3-x)O(4) and Cr(x)Fe(3-x)O(4) (0Cr(3+) and the influence of the three ions on sample thermostability is Zn(2+)>Ni(2+)>Cr(3+).