KDM3A promotes oral squamous cell carcinoma cell proliferation and invasion via H3K9me2 demethylation-activated DCLK1.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a frequently-diagnosed malignancy with high potential for proliferation and invasion. Histone methylation is known as a crucial mechanism that regulates pathological processes in various cancers, including OSCC. OBJECTIVE: This study sought to delve into the molecular mechanism of lysine demethylase 3 A (KDM3A) in OSCC cell proliferation and invasion. METHODS: Expression levels of KDM3A, lysin-9 of di-methylated histone H3 (H3K9me2), and doublecortin-like kinase 1 (DCLK1) in cells were determined by reverse-transcription quantitative polymerase chain reaction or Western blot analysis. Cell proliferation and invasion were evaluated by cell counting kit-8, colony formation, and Transwell assays. The enrichment of KDM3A and H3K9me2 on the DCLK1 promoter was determined by chromatin immunoprecipitation assay. The functional rescue experiment was performed with DCLK1 overexpression vector and si-KDM3A in CAL-27 and SCC-9 cells. RESULTS: KDM3A was elevated in OSCC cells. KDM3A knockdown suppressed OSCC proliferation and invasion, along with increased H3K9me2 level in OSCC cells. KDM3A and H3K9me2 were enriched on the DCLK1 promoter and inhibiting H3K9me2 improved DCLK1 expression levels. DCLK1 overexpression neutralized the inhibition of KDM3A knockdown on OSCC proliferation and invasion. CONCLUSIONS: KDM3A facilitated OSCC proliferation and invasion by eliminating H3K9me2 to upregulate DCLK1 expression levels.

AGEs-RAGE axis mediates myocardial fibrosis via activation of cardiac fibroblasts induced by autophagy in heart failure.

NEW FINDINGS: What is the central question of this study? Does the advanced glycation end products (AGEs)-receptor for advanced glycation end products (RAGE) axis mediate myocardial fibrosis in heart failure? What is the main finding and its importance? The AGEs-RAGE axis is involved in the pathogenesis of myocardial fibrosis through activation of cardiac fibroblasts induced by autophagy in heart failure. By suppression of cardiac fibroblast activation, inhibition of the AGEs-RAGE axis attenuates cardiac dysfunction and myocardial fibrosis in mice with transverse aortic constriction. ABSTRACT: Heart failure is the end stage of cardiovascular disease and is a critical medical condition that poses an important therapeutic challenge for physicians owing to its high morbidity and mortality. Myocardial fibrosis is part of the remodelling process that occurs in heart failure. Many studies have shown that advanced glycation end products (AGEs) and receptor for advanced glycation end products (RAGE) are implicated in fibrosis and autophagy, but the mechanism remains unclear. In this study, we elucidated the mechanism by which the AGEs-RAGE axis mediates activation of cardiac fibroblasts (CFs) in heart failure. We used C57BL/6J wild-type (WT) mice to establish a model of heart failure by transverse aortic constriction (TAC). After 6 weeks of treatment, relevant indicators were detected. In mice subjected to TAC, AGEs were upregulated compared with sham-operated mice. Inhibition of RAGE resulted in functional cardiac protection, with reduced hypertrophy and fibrosis in mice after TAC. Of note, autophagy mediated the activation of CFs that transformed to myofibroblasts and contributed to fibrosis. In vitro, CFs were obtained from neonatal Sprague-Dawley rats and treated with AGEs, bovine serum albumin and short hairpin RNA (shRNA) for RAGE, in order to verify the results obtained in vivo. These results suggest that the AGEs-RAGE axis is involved in the pathogenesis of myocardial fibrosis in heart failure through CF activation induced by autophagy. Inhibition of the AGEs-RAGE axis attenuates cardiac dysfunction and myocardial fibrosis in mice with TAC by suppressing CF activation.

Development of gene signature and nomogram for diagnosis and prognosis of oral carcinoma.

OBJECTIVE: The present study aimed to construct prognostic and diagnostic gene signatures and nomograms in oral carcinoma. DESIGN: Differentially expressed genes between oral carcinoma and normal samples were analyzed based on data from The Cancer Genome Atlas and Gene Expression Omnibus. Cox univariate and multivariate analyses and lasso analysis were conducted to construct the prognostic gene signature. Moreover, logit regression analysis was performed to establish the diagnostic model. Receiver operating characteristic curve, nomogram, calibration curve were utilized for validating the performance of the signature and the diagnostic model. RESULTS: A new 4-gene (ZNF114, AREG, GRB14, and DDIT4) prognostic signature was successfully constructed. Samples with higher risk score exhibited a worse overall survival than that with lower risk score in both The Cancer Genome Atlas cohort and GSE41613. Moreover, the receiver operating characteristic curve revealed a good performance of this signature in the prediction of survival. Furthermore, the signature was an independent prognosticator for oral carcinoma patients. Also, the nomogram and calibration curves showed a good clinical prediction effect. What's more, a diagnostic model including ZNF114, GRB14, and DDIT4 was constructed and presented a good performance in distinguishing oral carcinoma and normal samples. CONCLUSIONS: The present study developed a new 4-gene prognostic signature and nomogram, and constructed a 3-gene diagnostic model and nomogram to help in the diagnosis and prognosis of oral carcinoma.

Effects of Hydrothermal and Microwave Dual Treatment and Zein on the Enzymolysis of High Amylose Corn Starch.

Resistant starch (RS) type 2-high-amylose corn starch (HACS) was subjected to simultaneous hydrothermal (25% moisture content, 90 °C for 12 h) and microwave (35% moisture content, 40 W/g microwaving for 4 min) treatment and zein (at a zein to treated starch ratio of 1:5, 50 °C for 1 h) to improve its resistance to enzymolysis. Scanning electron microscopy (SEM) highlighted the aggregation and adhesion of the composite. The average particle size of the composite (27.65 µm) was exceeded that of both the HACS (12.52 µm) and the hydrothermal and microwave treated HACS (hydro-micro-HACS) (12.68 µm). The X-ray diffraction results revealed that the hydro-micro-HACS and composite remained B-type, while their crystallinity significantly decreased to 16.98% and 12.11%, respectively. The viscosity of the hydro-micro-HACS and composite at 50 °C was 25.41% and 35.36% lower than that of HACS. The differential scanning calorimetry (DSC) results demonstrated that the composite displayed a new endothermic peak at 95.79 °C, while the weight loss rate and decomposition temperature were 7.61% and 2.39% lower than HACS, respectively. The RS content in HACS, the hydro-micro-HACS, and composite was 47.12%, 57.28%, and 62.74%, respectively. In conclusion, hydrothermal and microwave treatment combined with zein provide an efficient physical strategy to enhance the RS type 2-HACS.

DNA repair genes are associated with tumor tissue differentiation and immune environment in lung adenocarcinoma: a bioinformatics analysis based on big data.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common type of lung cancer. DNA repair genes (DRGs) is important in lung cancer. The relationship between the immune environment and the expression levels of DRGs in LUAD remains unclear. The purpose of this study is to assess the relationship between DRGs and the immune environment and clinical characteristics of LUAD. METHODS: Data of 169 LUAD cases were obtained from cbioportal. The RNA-seq data came from the The Cancer Genome Atlas (TCGA) database. We collected DRGs from the Reactom database (KW0037, Reactom.org). The 302 genes expressed in each sample were analyzed by hierarchical clustering and grouped using the Gene Cluster 3.0 program. The Java Treeview program was used to generate heat maps of cluster indications and tumor staging patterns. GraphPad Prism 8 was used to draw survival curves and compare overall survival (OS). For single genes, an OS difference analysis between low and high expression populations was performed in GraphPad Prism 8. RESULTS: Matrix clustering showed no difference in the prognosis of the two clusters. The comparison of subgroups showed that Subcluster 1 (SC1) had the best prognosis, and Subcluster 2 (SC2) had the worst. There was a significant difference in tumor grades between Cluster 1 and Cluster 2 (P=0.01). There were significant differences in smoking status, histological grade and adenocarcinoma subtype among subgroups. In Subcluster 3 (SC3), the proportion of poorly differentiated cases was highest. Immunological index analysis showed that there were significant differences between Cluster 1 and Cluster 2 in interferon, macrophages, monocytes, neutrophils, natural killer (NK) cells, and T cells. Tumor purity, interferon, macrophages, monocytes, neutrophils, NK cells, T cells, translation, and proliferation all showed significant differences between subgroups. In SC2, the proliferation index increased (0.082 vs. 0.070); the protein translation index decreased (0.134 vs. 0.137); and the interferon level increased (0.099 vs. 0.097). In SC3, the proliferation index decreased (0.076 vs. 0.071); the protein translation index decreased (0.140 vs. 0.136); and the level of neutrophils increased (0.083 vs. 0.086). CONCLUSIONS: The differences of DRGs in LUAD are related to tissue differentiation and immune indicators but not to prognosis.

[Effect of moxibustion on postpartum urodynamics and pelvic floor function in puerperal women].

OBJECTIVE: To observe the effect of moxibustion on postpartum urodynamics and recovery of pelvic floor function based on the pelvic floor muscle function training. METHODS: A total of 150 puerperal women were randomly divided into an observation group (75 cases, 15 cases dropped off) and a control group (75 cases, 15 cases dropped off). The control group was treated with pelvic floor muscle function training, twice a day. Based on the treatment in the control group, the observation group was treated with Baixiao moxibustion at Qihai (CV 6), Guanyuan (CV 4), Sanyinjiao (SP 6) and Zusanli (ST 36), twice a week. The treatment started on the 42nd day after delivery, totaling for 12 weeks. The urodynamic indexes (functional urethral length [FUL], stress leak point pressure [SLPP], maximum urethral closure pressure [MUCP], bladder compliance [BC], maximum urethral pressure [MUP], detrusor pressure at maximum urinary flow rate [Pdet Qmax]), the international consultation on incontinence questionnaire-urinary incontinence-short form (ICIQ-UI-SF) score and vaginal muscle voltage were observed before and after treatment. The prolapse rate of pelvic floor organ and normal rate of pelvic floor muscle strength of the two groups were recorded after treatment. RESULTS: Compared before treatment, the levels of FUL, MUCP, BC, Pdet Qmax and SLPP in the observation group after treatment were increased (P<0.05), while the FUL and SLPP in the control group after treatment were increased (P<0.05). After treatment, the levels of FUL, SLPP, MUCP and BC in the observation group were higher than those in the control group (P<0.05). Compared before treatment, the ICIQ-UI-SF scores in the two groups after treatment were decreased (P<0.01), and the vaginal muscle voltage was increased (P<0.01). After treatment, the ICIQ-UI-SF score in the observation group was lower than that in the control group (P<0.01), and the vaginal muscle voltage in the observation group was higher than that in the control group (P<0.01). The prolapse rate of pelvic floor organ was 6.7% (4/60) in the observation group, which was lower than 20.0% (12/60) in the control group (P<0.05). The normal rate of pelvic floor muscle strength in the observation group was 88.3% (53/60), which was higher than 65.0% (39/60) in the control group (P<0.05). CONCLUSION: The moxibustion combined with pelvic floor muscle function training could improve postpartum urodynamics and pelvic floor muscle strength.

Analysis of Molecular Mechanism of YiqiChutan Formula Regulating DLL4-Notch Signaling to Inhibit Angiogenesis in Lung Cancer.

In order to explore the specific mechanism of YiqiChutan formula (YQCTF) in inhibiting the angiogenesis of lung cancer and its relationship with delta-like ligand 4- (DLL4-) Notch signaling, 30 healthy BALB/c-nu/nu rats were selected and divided into three groups: A549 group (implanted with lung adenocarcinoma cell line A549), NCI-H460 group (implanted with human lung large-cell carcinoma cell line NCI-H460), and NCI-H446 group (implanted with human lung small cell carcinoma cell line NCI-H446) for constructing lung cancer transplanted tumor models. After modeling, the group treated with normal saline was taken as control group, 200 mg/kg of YQCTF was adopted for intervention, and the tumor volume and growth inhibition rate were compared with the vascular targeted inhibitor Sorafenib. HE staining, CD31 fluorescent antibody staining, and microelectron microscopy were adopted to observe the neovascular endothelial cells of the transplanted tumor. The expression of VEGF, HIF-1α, DLL4, and Notch-1 in the transplanted tumors in each group was detected by Western blot and RT-PCR at the protein level or mRNA level. Compared with the control group, the YQCTF-treated group had obvious inhibitory effect on lung cancer transplanted tumor and lung cancer angiogenesis. In the YQCTF-treated group, the density of angiogenesis decreased significantly and the vascular lumen structure also decreased, and the expression levels of VEGF, HIF-1α, DLL4, and Notch-1 in the YQCTF-treated group were all lower than those in the control group. YQCTF could inhibit the growth of lung cancer transplanted tumor through antiangiogenesis, and it could also reduce the amount of angiogenesis in lung cancer transplanted tumor. In addition, the generation of lumen structure was also hindered, which was realized through the VEGF signaling pathway and DLL4-Notch signaling pathway.