DNA methylation heterogeneity attributable to a complex tumor immune microenvironment prompts prognostic risk in glioma.

Gliomas are malignant tumours of the human nervous system with different World Health Organization (WHO) classifications, glioblastoma (GBM) with higher grade and are more malignant than lower-grade glioma (LGG). To dissect how the DNA methylation heterogeneity in gliomas is influenced by the complex cellular composition of the tumour immune microenvironment, we first compared the DNA methylation profiles of purified human immune cells and bulk glioma tissue, stratifying three tumour immune microenvironmental subtypes for GBM and LGG samples from The Cancer Genome Atlas (TCGA). We found that more intermediate methylation sites were enriched in glioma tumour tissues, and used the Proportion of sites with Intermediate Methylation (PIM) to compare intertumoral DNA methylation heterogeneity. A larger PIM score reflected stronger DNA methylation heterogeneity. Enhanced DNA methylation heterogeneity was associated with stronger immune cell infiltration, better survival rates, and slower tumour progression in glioma patients. We then created a Cell-type-associated DNA Methylation Heterogeneity Contribution (CMHC) score to explore the impact of different immune cell types on heterogeneous CpG site (CpGct) in glioma tissues. We identified eight prognosis-related CpGct to construct a risk score: the Cell-type-associated DNA Methylation Heterogeneity Risk (CMHR) score. CMHR was positively correlated with cytotoxic T-lymphocyte infiltration (CTL), and showed better predictive performance for IDH status (AUC = 0.96) and glioma histological phenotype (AUC = 0.81). Furthermore, DNA methylation alterations of eight CpGct might be related to drug treatments of gliomas. In conclusion, we indicated that DNA methylation heterogeneity is associated with a complex tumour immune microenvironment, glioma phenotype, and patient's prognosis.

Exploring the tumor microenvironment: Chemokine-related genes and immunotherapy/chemotherapy response in clear-cell renal cell carcinoma.

BACKGROUND: The treatment of clear-cell renal cell carcinoma (ccRCC) remains challenge. Chemokines laid impact on the proliferation and metastasis of cancer cells. The objective was to identify the chemokine-related genes and construct a prognostic model for ccRCC. METHODS: Bulk transcriptomic data (n = 531), single-cell RNA sequencing (scRNA-seq) dataset GSE159115, and other validation cohorts were acquired from the Cancer Genome Atlas Program (TCGA) and GEO databases. All clustering analysis was conducted by Seurat R package. Gene set enrichment analysis (GSEA), immune infiltration analysis, single nucleotide variations (SNV) analysis, and predictive response analysis of immunotherapy/chemotherapy were conducted. 786-O and A498 cell lines were cultured and applied into CCK-8, Western blot, and RT-qPCR kits. RESULTS: Univariate Cox analysis was used to screen out chemokine-related genes related to survival. ZIC2, SMIM24, COL7A1, IGF2BP3, ITPKA, ADAMTS14, CYP3A7, and AURKB were identified and applied for the construction of the prognostic model. High-risk group had a poorer prognosis than the low-risk group in each dataset. Memory CD8+ T cells, macrophages, and memory B cells were higher in the high-risk group, while the content of basophils was higher in the low-risk group. Bortezomib_1191, Dactinomycin_1911, Docetaxel_1007, and Daporinad_1248 were more sensitive to high-risk groups than low-risk groups. Moreover, we found that IGF2BP3 significantly elevated in both 786-O and A498 cell lines resistance to sunitinib. Knockdown of IGF2BP3 markedly reduced ccRCC cell migration and viability. CONCLUSION: Our study has yielded a novel prognostic model of chemokine-related genes based on comprehensive transcriptional atlas of ccRCC patients, shedding light on the significant impact of the tumor microenvironment on biology and immunotherapy response of ccRCC. We identified IGF2BP3 as a pivotal regulator in regulating ccRCC resistance to sunitinib.

Differential expression of ASIP transcripts reveals genetic mechanism underpinning black-tail independence from body plumage in yellow-bodied chickens.

The genetic foundation of chicken body plumage color has been extensively studied. However, little attention has been paid to the inheritance patterns and molecular mechanisms underlying the formation of distal feather colors (tail and wingtip). Differences in these colors are common; for example, the Chinese Huiyang Beard chicken has black tail feathers, but yellow body plumage. Here, the hybrid offspring of Huiyang Beard and White Leghorn chickens were used to study the inheritance patterns of tail-feather color. The expression levels of pigment genes in differently colored feather follicles were analyzed using quantitative real-time PCR. The results showed that genetic regulation of tail-feather color was independent of body-plumage color. The Dominant White locus inhibited eumelanin synthesis in tail feathers without affecting the formation of yellow body plumage, whereas the Silver locus had the opposite effect. The expression of agouti signaling protein (ASIP) gene class 1 transcripts was significantly lower in black tail-feather follicles than in yellow body follicles, whereas tyrosinase-related protein 1 (TYRP1) gene expression was significantly higher in black tail feathers. These differentially expressed genes were confirmed to exert an effect on eumelanin and pheomelanin formation in feathers, thus influencing the regulation of chicken tail-feather color. In conclusion, this study lays the foundation for further research on the genetic mechanisms of regional differences in feather color, contributing to a better understanding of plumage pigmentation in chickens.

TIL-Derived CAR T Cells Improve Immune Cell Infiltration and Survival in the Treatment of CD19-Humanized Mouse Colorectal Cancer.

Chimeric antigen receptor-engineered T cells (CAR Ts) targeting CD19 have shown unprecedented prognosis in treating hematological cancers. However, the lack of a tumor-specific antigen as the target and an inhospitable tumor environment limit the clinical application of CAR T in solid tumors. Tumor-infiltrating T lymphocytes (TIL) exhibit diverse T cell receptor clonality and superior tumor-homing abilities. Therefore, in our study, human CD19-target TIL CAR-Ts armed with CD3ζ and 4-1BB signaling domains were constructed. Mouse colorectal cancer CT26 cells expressing human CD19 (hCD19+-CT26) were developed to assess the anti-tumor activity of TIL CAR-T cells, both in vitro and in vivo. Compared with splenic CAR T adoptive transfer, TIL CAR-T administration showed superior tumor suppression ability in hCD19+-CT26 tumor-bearing mice. Furthermore, more T cells were found at the tumor site and had lower exhaustion-related inhibitory receptor (T cell immunoglobulin and mucin domain-containing protein 3, Tim3) expression and higher immune memory molecule (CD62L) expression. Overall, we provided an artificial tumor-specific antigen in solid tumors and demonstrated that combined CAR-expressing TIL-Ts (TIL CAR-Ts) exhibited strong anti-tumor activity, with improved T cell infiltration and immune memory. Our humanized tumor antigen presented platform of mice suggests that TIL CAR-T-based adoptive therapy could be a promising strategy for solid cancer treatment.

Betulinic Acid Attenuates Osteoarthritis via Limiting NLRP3 Inflammasome Activation to Decrease Interleukin-1ß Maturation and Secretion.

Introduction: Osteoarthritis (OA) is the most common degenerative joint disorder. Prior studies revealed that activation of NLRP3 inflammasome could promote the activation and secretion of interleukin-1ß (IL-1ß), which has an adverse effect on the progression of OA. Betulinic acid (BA) is a compound extract of birch, whether it can protect against OA and the mechanisms involved are still unknown. Materials and Methods: In vivo experiments, using gait analysis, ELISA, micro-CT, and scanning electron microscopy (SEM), histological staining, immunohistological (IHC) and immunofluorescence (IF) staining, and atomic force microscopy (AFM) to assess OA progression after intraperitoneal injection of 5 and 15 mg/kg BA in an OA mouse model. In vitro experiments, caspase-1, IL-1ß, and the N-terminal fragment of gasdermin D (GSDMD-NT) were measured in bone marrow-derived macrophages (BMDMs) by using ELISA, western blot, and immunofluorescence staining. Results: We demonstrated that OA progression can be postponed with intraperitoneal injection of 5 and 15 mg/kg BA in an OA mouse model. Specifically, BA postponed DMM-induced cartilage deterioration, alleviated subchondral bone sclerosis, and relieved synovial inflammation. In vitro studies, the activated NLRP3 inflammasome produces mature IL-1ß by facilitating the cleavage of pro-IL-1ß, and BA could inhibit the activation of NLRP3 inflammasome in BMDMs. Conclusions: Taken together, our analyses revealed that BA attenuates OA via limiting NLRP3 inflammasome activation to decrease the IL-1ß maturation and secretion.

Effects of Glycine Supplementation in Drinking Water on the Growth Performance, Intestinal Development, and Genes Expression in the Jejunum of Chicks.

Glycine, the most basic amino acid found in nature, is considered an essential amino acid for chicks. However, the precise understanding of high concentrations of glycine's significance in promoting the growth performance of chicks, as well as its impact on intestinal development, re-mains limited. Consequently, the objective of this study was to investigate the effects of glycine supplementation in drinking water on growth performance, intestine morphology, and development in newly hatched chicks. In this study, 200 newly born chicks were selected and pro-vided with a supplementation of 0.5%, 1%, and 2% glycine in their drinking water during their first week of life. The results revealed that glycine supplementation in drinking water could significantly increase the average daily gain of chicks from days 7 to 14. Furthermore, a significant difference was observed between the group supplemented with 1% glycine and the control group. Concurrently, this glycine supplementation increased the villus height and the ratio of the villus height to crypt depth in jejunum on both day 7 and day 14. Glycine supplementation in drinking water significantly affected the mRNA expression level of the ZO-1, GCLM, and rBAT genes in jejunum, which may have certain effects on the mucosal immune defense, cellular antioxidant stress capacity, and amino acid absorption. Overall, the findings of this study indicate that glycine supplementation in drinking water can enhance the growth performance of chicks and promote their intestine development.

Chicken LEAP2 Level Substantially Changes with Feed Intake and May Be Regulated by CDX4 in Small Intestine.

Ghrelin O-acyltransferase (GOAT), ghrelin, and GHSR have been reported to play important roles that influence feed intake in mammals. LEAP2, an endogenous antagonist of GHSR, plays an important role in the regulation of feed intake. However, chicken ghrelin has also been reported to have an inhibitory effect on feed intake. The role of the GOAT-Ghrelin-GHSR-LEAP2 axis in chicken-feed intake remains unclear. Therefore, it is necessary to systematically evaluate the changes in the tissue expression levels of these genes under different energy states. In this study, broiler chicks in different energy states were subjected to starvation and feeding, and relevant gene expression levels were measured using quantitative real-time PCR. Different energy states significantly modulated the expression levels of LEAP2 and GHSR but did not significantly affect the expression levels of GOAT and ghrelin. A high expression level of LEAP2 was detected in the liver and the whole small intestine. Compared to the fed group, the fasted chicks showed significantly reduced LEAP2 expression levels in the liver and the small intestine; 2 h after being refed, the LEAP2 expression of the fasted chicks returned to the level of the fed group. Transcription factor prediction and results of a dual luciferase assay indicated that the transcription factor CDX4 binds to the LEAP2 promoter region and positively regulates its expression. High expression levels of GHSR were detected in the hypothalamus and pituitary. Moreover, we detected GHSR highly expressed in the jejunum-this finding has not been previously reported. Thus, GHSR may regulate intestinal motility, and this aspect needs further investigation. In conclusion, this study revealed the function of chicken LEAP2 as a potential feed-intake regulator and identified the potential mechanism governing its intestine-specific expression. Our study lays the foundations for future studies on avian feed-intake regulation.

Wheel-Running Exercise Protects Ovariectomized Mice from Bone Loss via IFN-γ-Mediated Suppression of the NF-κB and MAPK Pathways.

Physical exercise is recommended as a preventative approach for osteoporosis; however, the effect of physical exercise on bone mass remains controversial. Additionally, the immune regulation of physical exercise on bone mass remains unclear. To determine whether wheel-running (WR) exercise contributes to improving bone mineral density (BMD) and investigate the involved immune mechanism, ovariectomized (OVX) and sham-operated mice were treated with 8 weeks of WR exercise. The distal femurs of the mice were sequentially scanned, reconstructed, and analyzed using microcomputed tomography and related software to assess BMD and bone microarchitecture. Flow cytometry assays were applied to investigate alterations in immune cells and inflammatory cytokines. In vitro, osteoclast differentiation was conducted to determine the effect of IFN-γ on osteoclastogenesis and the underlying mechanism. As a result, trabecular parameters were decreased in the OVX mice compared with the sham group. However, WR exercise significantly improved the deterioration in the bone microarchitecture of the OVX mice with an increase of 60.00% in BMD, 55.18% in bone volume, 66.67% in trabecular number, 32.52% in trabecular thickness, and a decrease of 19.44% in trabecular separation. Similarly, WR exercise increased the proportion of CD8+ T cells from 7.26 ± 1.71% to 10.23 ± 1.35% in the spleen and from 1.62 ± 0.54% to 2.38 ± 0.43% in the bone marrow of the OVX mice (P < 0.05). The expression of IFN-γ was also increased in the OVX + WR mice compared with the OVX mice (1.65 ± 0.45% vs. 2.26 ± 0.34%, P < 0.05). In vitro studies demonstrated an inhibitory effect of IFN-γ on osteoclastogenesis in a dose- and time-dependent manner. Meanwhile, the classical NF-κB and MAPK pathways were found to be critical in IFN-γ-mediated inhibition of osteoclast differentiation. In conclusion, our study discovered that WR exercise rescued bone loss in the OVX mice in an IFN-γ-mediated immunomodulatory manner. After WR exercise, IFN-γ expression was restored by activated CD8+ T cells, consequently leading to the inhibition of osteoclastogenesis and the recovery from bone loss through the NF-κB and MAPK pathways.

Bone Mineral Density and Related Scores in Parkinson's Disease: A Systematic Review and Meta-Analysis.

BACKGROUND: Parkinson's disease (PD) is the second most common degenerative neurologic disorder in older adults, and increasing attention has been paid to bone health in PD. Although several studies have shown that patients with PD have a lower bone mineral density (BMD) than do non-PD controls, there have been no systematic reviews in recent years. METHODS: PubMed, Medline, and Web of Science were used to search relevant studies up to May 2020. BMD, BMD T score, and BMD Z score of patients with and without PD were statistically analyzed. Meta-analysis was conducted using Review Manager version 5.3. RESULTS: This meta-analysis included 17 studies comprising 10,289 individuals. In the meta-analysis, adults with PD had lower total body, total hip, total radius, lumbar spine, total femur, femur neck, right-hand, and left-hand BMD than did non-PD controls. The T score of total body BMD, total hip BMD, total radius BMD, lumbar spine BMD, L1-L4 spine BMD, total femur BMD, and femur neck BMD in adults with PD were lower than those in non-PD controls. Futhermore, the Z score of total body BMD, total hip BMD, total radius BMD, lumbar spine BMD, L1-L4 spine BMD, and femur neck BMD was lower in adults with PD than in non-PD controls. CONCLUSIONS: Patients with PD had a lower BMD, BMD T score, and BMD Z score compared with non-PD controls. Therefore, clinicians should routinely monitor BMD of patients with PD to prevent falling and fragility fractures in older adults and optimize BMD before surgical treatment of severe spinal deformity caused by PD.

Etoricoxib decreases subchondral bone mass and attenuates biomechanical properties at the early stage of osteoarthritis in a mouse model.

Etoricoxib, a selective Cyclooxygenase-2 (COX-2) inhibitor, is commonly used in osteoarthritis (OA) for pain relief, however, little is known about the effects on subchondral bone. In the current study, OA was induced via destabilization of the medial meniscus (DMM) in C57BL/6 mice. Two days after surgery, mice were treated with different concentrations of Etoricoxib. Four weeks after treatment, micro computed tomography (Micro-CT) analysis, histological analysis, atomic force microscopy (AFM) analysis, and scanning electron microscopy (SEM) were performed to evaluate OA progression. We demonstrated that Etoricoxib inhibited osteophyte formation in the subchondral bone. However, it also reduced the bone volume fraction (BV/TV), lowered trabecular thickness (Tb.Th), and more microfractures and pores were observed in the subchondral bone. Moreover, Etoricoxib reduced the elastic modulus of subchondral bone. Exposure to Etoricoxib further increased the empty/total osteocyte ratio of the subchondral bone. Etoricoxib did not show significant improvement in articular cartilage destruction and synovial inflammation in early OA. Together, our observations suggested that although Etoricoxib can relieve OA-induced pain and inhibit osteophyte formation in the subchondral bone, it can also change the microstructures and biomechanical properties of subchondral bone, promote subchondral bone loss, and reduce subchondral bone quality in early OA mice.

Guanidine sulfate-assisted synthesis of hexagonal WO3 nanoparticles with enhanced adsorption properties.

Large surface area hexagonal phase WO3 (h-WO3) nanowires were synthesized by a hydrothermal route with the assistance of C2H12N6O4S. They were characterized by XRD, SEM, TEM, BET, FT-IR and XPS. It is shown that C2H12N6O4S not only acts as a stabilizer to facilitate the generation of a metastable hexagonal phase, but also functions as a structure directing agent to assist the construction of nanowires. The obtained h-WO3 possesses a large specific surface area and numerous adsorption functional groups such as -OH groups. These characteristics result in an excellent adsorption performance for the removal of strontium from acidic aqueous solutions. A maximum adsorption capacity of 52.93 mg g(-1) was achieved on the h-WO3 prepared in the presence of C2H12N6O4S. This value is almost two times higher than that of bare h-WO3 (no C2H12N6O4S). The effects of pH, contact time, initial Sr(2+) concentration and ion strength on Sr(2+) removal from the solution by h-WO3 were systematically investigated. The adsorption mechanism involving the combination of electrostatic attraction and ion exchange for the adsorption of Sr(2+) is proposed. Based on our results, h-WO3 with high adsorption capacity and good surface characteristics exhibits great potential for the removal of Sr(2+) from radioactive wastewater.

Labeling conditions, in vitro properties and biodistributions of various Sn-labeled complexes.

Conditions for preparing Sn-EDTMP, Sn-DTPMP, Sn-TTHMP, Sn-HEDTMP and Sn-DTPA in aqueous solution in open air environments, and their in vitro properties including adsorption on hydroxyapatite (HA) and collagen (I) and binding to bovine serum albumin (BSA) were studied using (117m)Sn and 113Sn as tracers. Biodistributions of SnO2.xH2O.yEDTMP, SnO2.xH2O.yDTPMP, SnO2.xH2O.yTTHMP, SnO2.xH2O.yHEDTMP, SnO2.xH2O.yDTPA in normal mice were also tested. Based on the above experiments, the relationship between in vitro biochemical properties and biodistributions of these SnO2.xH2O.yLigands was investigated. The results show that Sn(IV)-Ligands are prone to hydrolysis into SnO2.xH2O.yLigands in aqueous solutions in open air environments, especially when the ligand is DTPA, when the molar ratio of metal to ligand is higher than 1:200, or when the pH of the solution is higher than 10. The in vitro experiments show that all of the SnO2.xH2O.yLigands bind strongly to BSA, and the binding percentages of SnO2.xH2O.yLigands to BSA are much higher than those of the corresponding Sn(IV)-Ligands. The biodistribution data indicate that all of the SnO2.xH2O.yLigands locate mainly in bone with little uptake in liver. When the binding percentages of SnO2.xH2O.yLigands to BSA are similar, those SnO2.xH2O.yLigands with higher adsorption on HA and collagen (I) undergo lower liver uptake.