Effects of Cadmium Exposure on Oxidative Stress in Atherosclerotic Rats by Downregulating TopBP1 Expression to Induce Mitochondrial DNA Damage.

Objective: To explore the mechanism of the effect of cadmium exposure on TopBP1-induced mitochondrial DNA damage in atherosclerotic rats to affect oxidative stress. Methods: 50 rats were established atherosclerotic model, and they were divided into model control group (MC group), low-dose cadmium exposure group (LD group), medium-dose cadmium exposure group (MD group), high-dose cadmium exposure group (HD group), and positive control group, with 10 rats in each group. Rats in the LD group, MD group, and HD group were intraperitoneally injected with different doses of cadmium acetate solution for intervention, rats in the PC group were intraperitoneally injected with oxidized banking solution, and those in the MC group were injected with normal saline. 10 rats were taken as the normal control group (NC group). Human umbilical vein endothelial cells were taken for cell experiments, normal saline was added as the blank control group (group A), cadmium acetate solution was added (group B), oxidized bankning solution was added (Group C), and oxidized bankning solution and cadmium acetate solution were added (Group D). Western blot and fluorescence quantitative PCR were used to detect the protein and mRNA expressions respectively. ROS, MDA, and SOD were detected by ELISA, apoptosis of endothelial cells was detected by flow cytometry, and arterial plaque damage was observed by oil red O staining. Results: The relative expressions of TopBP, Bax, and Bcl-2 proteins in rat aortic tissues in each group were significantly different (all P < .05). The relative expressions of TopBP1 and Bcl-2 proteins in the aortic tissues of rats in NC group, MC group, LD group, MD group, HD group, and PC group decreased (all P < .05), while the relative expressions of Bax protein in those groups were increased (all P < .05). Similarly, the relative expression levels of Topbp1mRNA, BaxmRNA, and Bcl-2mRNA in the aortic tissues of rats in each group were significantly different (all P < .05). There were statistically significant differences in the expression levels of ROS, MDA, SOD, and mtDNA expression levels in the aortic tissues of rats in each group. There were statistically significant differences in TopBP1, Topbp1mRNA, and mtDNA among groups (all P < .05); while the relative expression of TopBP1 and Topbp1mRNA in groups A, B, C, and D decreased (all P < .05), the expression levels of mtDNA in those group increased (all P < .05), and the apoptosis rates of endothelial cells were also increased (all P < .05). Conclusion: Cadmium exposure can down-regulate the expression of TopBP1 in atherosclerotic rats, aggravate mitochondrial DNA damage, promote oxidative stress response, and then induce the development of atherosclerosis.

Upregulation of the Four and a Half LIM Domains 1 linked with familial venous dysplasia in a familial genetic examination.

BACKGROUND: This study aimed to analyze the mutation site in a family diagnosed with venous dysplasia to identify possible pathogenic genes. METHODS: A 15-year-old female presented with lower extremity venous tortuosity aggravated by ulceration. Only the young sister exhibited similar symptoms within the immediate family of the proband. Whole genome sequencing (WGS) was used to evaluate the mutation sites and chromosome copy number variations (CNV) within the family. The possible pathogenic genes located in the region with CNVs were identified, and the expression of the possible pathogenic genes was verified via quantitative polymerase chain reaction (Q-PCR) and western blotting (WB) analysis. In-vitro models were used to verify the role of possible pathogenic genes linked with the development of venous dysplasia. RESULTS: The high-resolution karyotype analysis of the chromosomes found no abnormalities. The results of the WGS indicated that the proband and her sister shared the CNV events, including a microdeletion on chromosomes X: 13580000-1358555000 and microduplications of chromosome X: 136055000-136290000, chromosome X: 136475000-13671000. The results of the Q-PCR and WB showed that FHL1 was highly expressed in the proband and her sister, indicating that mutations of the FHL1 may have an important role in the development of vein malformations. The results of the in vitro experiments showed that FHL1 overexpression could inhibit venous development. CONCLUSION: The CNV in the Xq26 region (136054501-136288300) was found to be linked with the development of venous malformations in this family. However, further studies are required to evaluate the genetic mechanisms involved in the development of venous malformations.

LIM Homeobox 2 Increases Adhesion-Regulating Molecule 1 Transcription to Facilitate the Pathological Progression of Oxidized Low-Density Lipoprotein-Stimulated Atherosclerotic Cell Models.

Endothelial-mesenchymal transition (EndMT) and endothelial cell apoptosis have been documented to have a role in atherosclerosis (AS) progression. To deepen knowledge in this aspect, our study investigated the effect of LIM homeobox 2 (LHX2) and adhesion-regulating molecule 1 (ADRM1) on EndMT and endothelial cell apoptosis in the oxidized low-density lipoprotein (ox-LDL) -stimulated AS cell model.Ox-LDL was utilized to treat human umbilical vein endothelial cells (HUVECs) for constructing an AS model in vitro, followed by measurement of LHX2 and ADRM1 expressions. Afterward, gain- and loss-of-function assays were performed in HUVECs, followed by detection of cell viability, invasion, migration, and apoptosis and the expression of inflammatory factors [tumor necrosis factor (TNF) -α, interleukin (IL) -1ß, and IL-6], EndMT-related proteins [CD31, vascular epithelium (VE) -cadherin, vimentin, α-smooth muscle actin (SMA), Snai1, Snai2, and Twist1], and the apoptotic protein cleaved caspase-3. Interactions between LHX2 and ADRM1 were analyzed with dual-luciferase reporter gene and chromatin immunoprecipitation assays.High levels of LHX2 and ADRM1 were observed in ox-LDL-induced HUVECs. In ox-LDL-treated HUVECs, LHX2, or ADRM1 knockdown promoted CD31 and VE-cadherin levels, viability, invasion, and migration and reduced apoptosis and the expressions of TNF-α, IL-1ß, IL-6, vimentin, α-SMA, Snai1, Snai2, Twist1, and cleaved caspase-3. Mechanistically, LHX2 bound to the ADRM1 promoter to promote ADRM1 transcription. Overexpression of ADRM1 annulled the aforementioned effects of LHX2 knockdown on ox-LDL-induced HUVECs.LHX2 facilitates the pathological progression of ox-LDL-stimulated AS cell models by increasing ADRM1 transcription.

Overexpression of tripartite motif-containing 47 (TRIM47) confers sensitivity to PARP inhibition via ubiquitylation of BRCA1 in triple negative breast cancer cells.

Triple-negative breast cancers (TNBC) frequently harbor defects in DNA double-strand break repair through homologous recombination (HR), such as BRCA1 dysfunction. However, less than 15% of TNBC patients were found to carry BRCA1 mutation, indicating that there are other mechanisms regulating BRCA1-deficient in TNBC. In the current study, we shown that overexpression of TRIM47 correlates with progression and poor prognosis in triple-negative breast cancer. Moreover, we demonstrated that TRIM47 directly interacts with BRCA1 and induces ubiquitin-ligase-mediated proteasome turnover of BRCA1, subsequently leads to a decrease of BRCA1 protein levels in TNBC. Moreover, the downstream gene expression of BRCA1, such as p53, p27, p21 was significantly reduced in the overexpression of TRIM47 cell lines but increased in TRIM47-deleted cells. Functionally, we found that overexpression of TRIM47 in TNBC cells confers an exquisite sensitivity to olaparib, an inhibitor of poly-(ADP-ribose)-polymerase (PARP), but TRIM47 inhibition significantly confers TNBC cells resistance to olaparib both in vitro and in vivo. Furthermore, we showed that overexpression of BRCA1 significant increase the olaparib resistance in TRIM47-overexpression-induced PARP inhibitions sensitivity. Taken together, our results uncover a novel mechanism for BRCA1-deficient in TNBC and targeting TRIM47/BRCA1 axis may be a promising prognostic factor and a valuable therapeutic target for TNBC.

CircSOD2 Contributes to Tumor Progression, Immune Evasion and Anti-PD-1 Resistance in Hepatocellular Carcinoma by Targeting miR-497-5p/ANXA11 Axis.

Circular RNAs (circRNAs) can function as functional molecules in hepatocellular carcinoma (HCC). Herein, circRNA superoxide dismutase 2 (circSOD2) was researched in HCC progression and immune system. The real-time polymerase chain reaction (qRT-PCR) was used for quantification of circSOD2, microRNA-497-5p (miR-497-5p) and Annexin A11 (ANXA11). Cell assays were performed by 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT) and colony formation assays for proliferation, flow cytometry for apoptosis and cell cycle, wound healing assay for migration and transwell assay for migration/invasion. ANXA11 and metastatic protein levels were measured by western blot. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to analyze target binding. CD8+ T cell immunity was assessed by Immunohistochemistry (IHC) assay, and the effect of circSOD2 on programmed cell death 1 (PD-1) immune checkpoint inhibitors (anti-PD-1) therapy was evaluated by mice xenograft assay. CircSOD2 was upregulated in HCC tissues and cells. Knockdown of circSOD2 resulted in HCC cell growth inhibition, apoptosis promotion, cell cycle arrest and metastasis suppression. Mechanically, circSOD2 promoted HCC development by acting as a miR-497-5p sponge and miR-497-5p played a tumor-inhibitory role in HCC cells by targeting ANXA11. Moreover, circSOD2 induced upregulation of ANXA11 expression by interacting with miR-497-5p. Also, the promoting effects of circSOD2 on immune evasion and anti-PD-1 resistance were related to miR-497-5p/ANXA11 axis. This study elucidated the pivotal function of circSOD2 in HCC progression and immunosuppression by mediating miR-497-6p/ANXA11 axis. CircSOD2/miR-497-5p/ANXA11 axis was a novel view of circRNA research in HCC.

Hepatic portal venous gas complication associated with the thoracic endovascular aortic repair for aortic dissection: a case report and literature review.

Aortic dissection (AD) is a serious disease with a higher mortality. The thoracic endovascular aortic repair (TEVAR) is a first line regimen for aortic dissection. Hepatic portal venous gas (HPVG) is a rare disease, and its definite mechanism is unknown. This is a rare association between the aortic and HPVG. In the present report, we present a case of thoracic aortic dissection, which was the type of Standford B by the computer tomography (CT) angiography, which implicated acute abdominal pain and abdominal distention after TEVAR and immediate abdominal CT shown hepatic portal venous gas (HPVG). The patient, who was treated with conservative treatment of gastrointestinal decompressing, fluid resuscitation, electrolyte replacement, anti-infection, anti-inflammation and anticoagulation, was recovered and discharged without abnormalities. This patient has been followed up for 5 years and has not experienced any physical discomfort related to HPVG. This is the first report that the aortic dissection patient implication with HPVG after thoracic endovascular aortic repair.

Deep Learning Reconstruction Algorithm-Based MRI Image Evaluation of Edaravone in the Treatment of Lower Limb Ischemia-Reperfusion Injury.

This research aimed to evaluate the therapeutic effect of edaravone on lower limb ischemia-reperfusion injury by MRI images of graph patch-based directional curvelet transform (GPBDCT), compression reconstruction algorithm. 200 patients with lower limb ischemia-reperfusion injury after replantation of severed limb were randomly divided into the observation group (edaravone treatment) and control group (Mailuoning injection treatment), with 100 cases in each group. MRI scanning and image processing using the GPBDCT algorithm were used to evaluate the therapeutic effect of the two groups of patients. The results showed that the signal noise ratio (SNR) (22.01), relative l 2 norm error (RLNE) (0.0792), and matching degree γ (0.9997) of the compression and reconstruction algorithm based on GPBDCT were superior to those of the conventional compression and reconstruction algorithm (P < 0.05). MRI examination showed that the decrease of bleeding signal after treatment in the observation group was superior to that in the control group. The levels of superoxide dismutase (SOD) (15 ± 2.02), malondialdehyde (MDA) (2.27 ± 1.02), B cell lymphoma-2 (Bcl-2) (8.5 ± 1.02), Bcl-2-associated X (Bax) (3.7 ± 0.42), and Caspase-3 protein (35.9 ± 5.42) in the observation group before and after treatment were significantly higher than those in the control group (P < 0.05). In conclusion, the GPBDCT-based compression reconstruction algorithm has a better effect on MRI image processing, and edaravone can better remove free radicals and alleviate apoptosis.

A combination of transcriptome and methylation analyses reveals the role of lncRNA HOTAIRM1 in the proliferation and metastasis of breast cancer.

Background: DNA methylation status is strongly associated with the prognosis of breast invasive carcinoma (BRCA). Elucidating the mechanisms underlying DNA methylation coupled with determining its biological function is imperative to the effective development of treatment and prevention strategies for breast cancer. Methods: We retrieved transcriptome and DNA methylation profiles of BRCA patients from The Cancer Genome Atlas (TCGA) database, then applied the "limma" package in R software to identify differentially expressed genes (DEGs) and aberrantly methylated genes. Next, we used the "MethylMix" package to screen for methylation-driven genes, and performed univariate and multivariate Cox regression analyses to determine the prognostic value of the methylation-driven genes and clinical characteristics. We validated these findings in 51 breast cancer tissues alongside 51 corresponding normal tissues. Furthermore, we used cell experiments to clarify the biological function and underlying molecular mechanisms of HOTAIRM1 in vitro. Results: A total of 25 methylation-driven genes were identified in the dataset. Results from univariate and multivariate Cox regression showed that SYN2, HOTAIRM1, BCAS1, and ALDOC were significantly associated with patient prognosis. Immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) results showed that the expression levels of SYN2 and HOTAIRM1 were negatively correlated with BRCA stage, whereas those of BCAS1 and ALDOC were positively correlated with BRCA stage. Results from in vitro experiments showed that knockdown HOTAIRM1 expression promoted breast cancer cells proliferation, clone formation, and invasion. Up-regulation of HOTAIRM1 inhibited breast cancer cells proliferation, clone formation, and invasion. Conclusions: In summary, low HOTAIRM1 expression is a significant prognostic factor for the survival of BRCA patients and thus could be a potential therapeutic target for the treatment of BRCA.

A Competing Endogenous RNA Network Reveals Novel lncRNA, miRNA and mRNA Biomarkers With Diagnostic and Prognostic Value for Early Breast Cancer.

BACKGROUND: This study aims to reveal early breast cancer (BC) specific competing endogenous RNA (ceRNA) network through the expression profiles of microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and mRNAs. METHODS: Based on The Cancer Genome Atlas (TCGA), we obtained the differentially expressed mRNAs, miRNAs, and lncRNAs (DEmRNAs, DEmiRNAs and DElncRNAs) between early BC and normal samples. The lncRNA-miRNA-mRNA interaction network was constructed using Cytoscape. Functional enrichment were performed using GeneCoDis3. The expression of selected genes were validated by qRT-PCR. Based on the published dataset, we validated the result of TCGA integration analysis. The diagnostic and prognostic value of candidate genes was evaluated by ROC curve analysis and survival analysis, respectively. RESULTS: Totally, 1207 DEmRNAs, 194 DElncRNAs and 37 DEmiRNAs were obtained. Functional enrichment analysis results showed that all of DEmRNAs were enriched in pathway of cytokine-cytokine receptor interaction, PPAR signaling pathway and pathways in cancer. The DEmRNA-DEmiRNA-DElncRNA interaction network in early BC was consisted of 23 DEmiRNAs, 95 DElncRNAs and 309 DEmRNAs. Among ceRNA network, IL-6-hsa-miR-182-5p-ADAMTS9-AS1 interactions, LIFR-hsa-miR-21-5p-ADAMTS9-AS1 interactions and MMP1/MMP11-hsa-miR-145-5p-CDKN2B-AS1 interactions were speculated to involve in the development of early BC. The qRT-PCR results were consistent with our integrated analysis. Except for ADAMTS9-AS1 and CDKN2B-AS1, expression of the others results in the Gene Expression Omnibus (GEO) dataset were generally consistent with TCGA integrated analysis. The area under curve (AUC) of the ADAMTS9-AS1, CDKN2B-AS1, IL-6, MMP11, hsa-miR-145-5p and hsa-miR-182-5p were 0.947, 0.862, 0.842, 0.993, 0.960 and 0.944, and the specificity and sensitivity of the 6 biomarkers were 83.4% and 95.6%, 72.2% and 90.3%, 80.1% and 74.3%, 96.2% and 96.5%, 90.1% and 92.3%, and 88.7% and 90.4%, respectively. In addition, IL-6 had potential prognostic value for early BC. CONCLUSION: These findings may provide novel insights into the lncRNA-miRNA-mRNA network and uncover potential therapeutic targets in early BC.

Hinokiflavone induces apoptosis and inhibits migration of breast cancer cells via EMT signalling pathway.

Hinokiflavone is a natural product, isolated from Selaginella P. Beauv, Juniperus phoenicea and Rhus succedanea. Even though hinokiflavone was reported to possess cytotoxicity to many cancer cells, and has potential in cancer treatment, the anti-proliferation and anti-metastasis efficacy of hinokiflavone on human breast cancer cells has not a further research. In this study, we investigated the anti-cancer activity of hinokiflavone in human breast cancer cells in vitro and in vivo. Hinokiflavone exhibited a time- and dose-dependent manner apoptosis induction by upregulating expression of Bax and downregulating Bcl-2 in breast cancer cells. Furthermore, hinokiflavone significantly inhibited the migration and invasion of breast cancer cells by impairing the process of epithelial-to-mesenchymal transition. In addition, the tumour growth was distinctly inhibited by treatment of hinokiflavone in a xenograft tumour mouse model of MDA-MB-231 cells. Immunohistochemical analysis of tumour sections showed that MMP-2+ cells and Ki-67+ cells were remarkably decreased in tumour tissues of mice after treatment of hinokiflavone, indicating that hinokiflavone inhibits not only proliferation but also metastasis of breast cancer cells. Our study suggested that hinokiflavone can be a potential drug to breast cancer. SIGNIFICANCE OF THE STUDY: Hinokiflavone significantly inhibited proliferation and induced apoptosis in breast cancer cells. In addition, hinokiflavone remarkably inhibited migration and invasion of breast cancer cells via EMT signalling pathway. It is worth noting that hinokiflavone possesses anti-tumour effect in tumour mouse xenograft model of breast cancer. Overall, our results indicated that hinokiflavone may be a potential anticancer drug for breast cancer treatment.

The molecular mechanism of breast cancer cell apoptosis induction by absent in melanoma (AIM2).

OBJECTIVE: To determine the molecular mechanism by which absent in melanoma 2 (AIM2) induces breast cancer cell apoptosis. METHODS: Establish Tet-Off(TM) model system to induce AIM2 expression in great quantities, MCF-7 tTA-AIM2 cells were the experimental group; MCF-7 tTA-Luc cells were the control group. The expression and subcellular localization of AIM2 in breast cancer cell lines were determined via Western Blotting. AIM2 protein expression was determined after the addition of interferon-γ (102 U/ml). Flow cytometry was used to analyze the effects of AIM2 on the cell cycle. Apoptosis detection was performed by staining with Annexin V-FITC and propidium iodide. Apoptosis mechanism was detected via Western Blotting. The XTT assay was used to analyze the effects of AIM2 on cell growth. RESULT: This experiment established Tet-Off guidance system. This system results which promotes AIM2 gene transcription and increased AIM2 protein expression. Four days after induction, AIM2 expression was detected. AIM2 expression increased with the number of days post-induction. AIM2 is present in cytoplasm and nuclei. Interferon-γ (102 U/ml) induced AIM2 protein expression and significantly increased AIM2 expression. AIM2 expression had no significant effect on the cell cycle, With the increase of Cdk2 expression induced by days were gradually increased, and Cdk4, Cyclin E expression was no significantly difference. AIM2 expression can significantly promote the apoptosis of breast cancer cells. Increased AIM2 expression can inhibit the expression of the anti-apoptotic protein Bcl-xL, increase the expression of the apoptosis proteins Bad and Bax, and activate caspases, resulting in cleavage of the DNA repair protein PARP. The XTT assay showed that AIM2 expression slows the rate of cell growth. CONCLUSION: In this breast cancer Tet-Off(TM) system, AIM2 was expressed in the cytoplasm and nucleus, stimulated the mitochondria to promote apoptosis, and influenced cell survival and proliferation.

miR-144 regulates transforming growth factor-ß1 iduced hepatic stellate cell activation in human fibrotic liver.

OBJECTIVES: Activation of hepatic stellate cells (HSCs) into collagen producing myofibroblasts is critical for pathogenesis of liver fibrosis. Transforming growth factor-ß1 (TGF-ß1) is one of the main profibrogenic mediators for HSC transdifferentiation. Recent studies have shown effect of microRNAs (miRNAs) on regulating TGF-ß1-induced HSC activation during liver fibrosis. Here, we aimed to explore the roles of miR-144 and miR-200c in human liver fibrosis. METHODS: Expression of TGF-ß1 was detected in 42 fibrotic and 18 normal human liver tissues by quantitative real time polymerase chain reaction (qRT-PCR) and immunohistochemistry, and its correlation with α-smooth muscle actin (α-SMA) was calculated. miR-144 and miR-200c expression level in fibrotic liver tissues were also detected by qRT-PCR. The correlation of TGF-ß1 expression with miR-200c and miR-144 in the fibrotic liver was analyzed. RESULTS: The results showed that TGF-ß1 expression was much higher in fibrotic liver than that in normal liver tissues (P<0.05). TGF-ß1 protein high expressing liver fibrosis showed α-SMA positive cells in the liver parenchyma indicating activated HSCs. Expression of TGF-ß1 in fibrotic liver was significantly correlated with α-SMA expression (R=0.633, P<0.001). Furthermore, miR-144 was less expressed in liver fibrosis (P<0.05) and was significantly correlated with expression of TGF-ß1 in fibrotic liver tissues (R=-0.442, P<0.01). However, miR-200c did not show significant difference between normal and fibrotic liver (P=0.48) and correlation with TGF-ß1 expression (R=0.106, P=0.51). CONCLUSION: All the results indicate that miR-144 can be a novel regulator of TGF-ß1-induced HSC activation during liver fibrosis.

Midazolam inhibits the proliferation of human head and neck squamous carcinoma cells by downregulating p300 expression.

The objective of this study was to investigate the mechanism of midazolam in inhibiting the proliferation of hypopharyngeal squamous carcinoma cells. Cultured FaDu cancer cells were treated with different concentrations of midazolam. MTT and BrdU incorporation assays were then used to evaluate cancer cell proliferation. The mRNA and protein levels of p300, a key factor involved in the tumorigenesis of numerous cancers, were measured with RT-PCR and Western blotting, respectively. Midazolam inhibited the expression of p300 and the proliferation of FaDu cells. Additionally, knockdown of p300 resulted in increased expression of p21 and p27 and decreased expression of p-Rb while inhibiting the proliferation of FaDu cells. Midazolam inhibits the proliferation of human head and neck squamous carcinoma cells by downregulating p300. Midazolam may be useful for the treatment of hypopharyngeal squamous cancers.

A rapidly progressive giant tumor with one year growth on breast: a rare size breast mass.

Primary rapidly progressive malignant tumor on breast is a very rare disease. We report a 62-year-old postmenopausal patient with history of one year breast lump and progressive increase in a period of two months. The giant breast tumor with a rare size about 7.5 × 7.0 cm, the skin of the breast to become resembling orange peel and metastasis to multiple organs. The histological sections of core needle biopsy confirmed the diagnosis of invasive adenocarcinoma. Hence, reduction surgery for local skin in the treatment of breast cancer increased the effectiveness of the docetaxel epirubicin (TE: docetaxel 75 mg/m(2), day 1; epirubicin 75 mg/m(2), day 1) chemotherapy by improving the patient's general condition, and not continue to increase was observed during follow-up.