The vitamin D receptor in osteoblastic cells but not secreted parathyroid hormone is crucial for soft tissue calcification induced by the proresorptive activity of 1,25(OH)2D3.

The vitamin D receptor (VDR) is expressed most abundantly in osteoblasts and osteocytes (osteoblastic cells) in bone tissues and regulates bone resorption and calcium (Ca) and phosphate (P) homeostasis in association with parathyroid hormone (PTH). We previously reported that near-physiological doses of vitamin D compounds suppressed bone resorption through VDR in osteoblastic cells. We also found that supra-physiological doses of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] induced bone resorption and hypercalcemia via VDR in osteoblastic cells. Here, we report that the latter, a proresorptive dose of 1,25(OH)2D3, causes soft tissue calcification through VDR in osteoblastic cells. High concentrations of vitamin D affect multiple organs and cause soft tissue calcification, with increases in bone resorption and serum Ca levels. Such a variety of symptoms is known as hypervitaminosis D, which is caused by not only high doses of vitamin D but also impaired vitamin D metabolism and diseases that produce 1,25(OH)2D3 ectopically. To clarify the biological process hierarchy in hypervitaminosis D, a proresorptive dose of 1,25(OH)2D3 was administered to wild-type mice in which bone resorption had been suppressed by neutralizing anti-receptor activator of NF-κB ligand (RANKL) antibody. 1,25(OH)2D3 upregulated the serum Ca x P product, concomitantly induced calcification of the aorta, lungs, and kidneys, and downregulated serum PTH levels in control IgG-pretreated wild-type mice. Pretreatment of wild-type mice with anti-RANKL antibody did not affect the down-regulation of PTH levels by 1,25(OH)2D3, but inhibited the increase of the serum Ca x P product and soft tissue calcification induced by 1,25(OH)2D3. Consistent with the effects of anti-RANKL antibody, VDR ablation in osteoblastic cells also did not affect the down-regulation of PTH levels by 1,25(OH)2D3, but suppressed the 1,25(OH)2D3-induced increase of the serum Ca x P product and calcification of soft tissues. Taken together with our previous results, these findings suggest that bone resorption induced by VDR signaling in osteoblastic cells is critical for the pathogenesis of hypervitaminosis D, but PTH is not involved in hypervitaminosis D.

Orthodontic-surgical treatment for severe skeletal class II malocclusion with vertical maxillary excess and four premolars extraction: A case report.

BACKGROUND: Patient satisfaction with facial appearance at the end of orthodontic camouflage treatment is very important, especially for skeletal malocclusion. This case report highlights the importance of the treatment plan for a patient initially treated with four-premolar-extraction camouflage, despite indications for orthognathic surgery. CASE SUMMARY: A 23-year-old male sought treatment complaining about his unsatisfactory facial appearance. His maxillary first premolars and mandibular second premolars had been extracted, and a fixed appliance had been used to retract his anterior teeth for two years without improvement. He had a convex profile, a gummy smile, lip incompetence, inadequate maxillary incisor inclination, and almost a class I molar relationship. Cephalometric analysis showed severe skeletal class II malocclusion (A point-nasion-B point = 11.5°) with a retrognathic mandible (sella-nasion-B point = 75.9°), a protruded maxilla (sella-nasion-A point = 87.4°), and vertical maxillary excess (upper incisor to palatal plane = 33.2 mm). The excessive lingual inclination of the maxillary incisors (upper incisor to nasion-A point line = -5.5°) was due to previous treatment attempts to compensate for the skeletal class II malocclusion. The patient was successfully retreated with decompensating orthodontic treatment combined with orthognathic surgery. The maxillary incisors were repositioned and proclined in the alveolar bone, the overjet was increased, and a space was created for orthognathic surgery, including maxillary impaction, anterior maxillary back-setting, and bilateral sagittal split ramus osteotomy to correct his skeletal anteroposterior discrepancy. Gingival display was reduced, and lip competence was restored. In addition, the results remained stable after 2 years. The patient was satisfied with his new profile as well as with the functional malocclusion at the end of treatment. CONCLUSION: This case report provides orthodontists a good example of how to treat an adult with severe skeletal class II malocclusion with vertical maxillary excess after an unsatisfactory orthodontic camouflage treatment. Orthodontic and orthognathic treatment can significantly correct a patient's facial appearance.

Massive Fabrication of Flexible, Form-Stable, and Self-Repairing Brine Phase Change Material Gels toward Smart Cold Chain Logistics.

Cold chain logistics plays an extremely important role in the storage and transportation of perishable products. Nowadays, phase change materials (PCMs) have been applied in emerging cold chain logistics to overcome the problems of low stability, high energy consumption, and high cost in mechanical refrigeration-based cold chain logistics. Mass production of high-performance phase change cold storage materials toward cold chain logistics is still a major challenge. Herein, self-repairing brine phase change gels (BPCMGs) massively fabricated by ionic cross-linking, covalent cross-linking, and hydrogen bond cross-linking are proposed. Brine containing 23.3% sodium chloride (NaCl) is selected as the phase change component because its phase change temperature is suitable for the cold storage demand of aquatic products. The proposed BPCMGs demonstrate superior thermophysical properties in terms of no phase separation, no supercooling, high form stability, high latent heat, high thermal conductivity, high cyclic stability, and high self-repairing rate. Meanwhile, the BPCMGs present high cost-effectiveness. Given these advantages, BPCMGs are utilized to assemble smart cold storage equipment for the storage and transportation of aquatic products. The cold storage time reaches 36.73 h for aquatic products when the stored cold energy is 364078 J. The location and temperature of the refrigerated products are monitored in real-time. The state-of-the-art BPCMGs provide diversified possibilities for the advanced smart cold chain.

Coral-like Fe-doped MoO2/C heterostructures with rich oxygen vacancies for efficient electrocatalytic N2 reduction.

Molybdenum (Mo) is one of the most important constituent elements in natural nitrogenase and theoretical calculation results show that Mo-based materials can be used as potential NRR electrocatalysts. The design of advanced catalysts with a special structure is very essential for promoting the development of electrocatalytic N2 into NH3. In this paper, Fe-doped MoO2/C heterostructured nanoparticles with rich oxygen vacancies (Vo) are designed and they exhibit highly efficient catalytic activity for artificial N2 fixation in neutral electrolytes under ambient conditions. The influence of the atomic ratio of the Fe source to the Mo source and the NaBH4 ethanol solution treatment on the structure and electrocatalytic performance are systematically investigated. The Vo-Fe-MoO2/C (1 : 50) catalyst with rich oxygen vacancies shows a satisfactory electrocatalytic N2 reduction reaction (e-NRR) activity in 0.1 M Na2SO4 with a high ammonia yield rate of 15.87 ± 0.3 µg h-1 mg-1 at -0.5 V versus the reversible hydrogen electrode (vs. the RHE) and a FE of 13.4% at -0.3 V (vs. the RHE). According to the results of DFT calculations, the active center of the electro-catalytic nitrogen reduction reaction is the molybdenum atom between the iron atom and the O vacancy. Oxygen vacancies can not only reduce the energy barrier of the RDS but also facilitate the desorption of ammonia and the first step hydrogenation of nitrogen. The doping of Fe will change the electronic state of the Mo atom in MoO2.

The APC/CTAD1-WIDE LEAF 1-NARROW LEAF 1 pathway controls leaf width in rice.

Leaf morphology is one of the most important features of the ideal plant architecture. However, the genetic and molecular mechanisms controlling this feature in crops remain largely unknown. Here, we characterized the rice (Oryza sativa) wide leaf 1 (wl1) mutant, which has wider leaves than the wild-type due to more vascular bundles and greater distance between small vascular bundles. WL1 encodes a Cys-2/His-2-type zinc finger protein that interacts with Tillering and Dwarf 1 (TAD1), a co-activator of the anaphase-promoting complex/cyclosome (APC/C) (a multi-subunit E3 ligase). The APC/CTAD1 complex degrades WL1 via the ubiquitin-26S proteasome degradation pathway. Loss-of-function of TAD1 resulted in plants with narrow leaves due to reduced vascular bundle numbers and distance between the small vascular bundles. Interestingly, we found that WL1 negatively regulated the expression of a narrow leaf gene, NARROW LEAF 1 (NAL1), by recruiting the co-repressor TOPLESS-RELATED PROTEIN and directly binding to the NAL1 regulatory region to inhibit its expression by reducing the chromatin histone acetylation. Furthermore, biochemical and genetic analyses revealed that TAD1, WL1, and NAL1 operated in a common pathway to control the leaf width. Our study establishes an important framework for understanding the APC/CTAD1-WL1-NAL1 pathway-mediated control of leaf width in rice, and provides insights for improving crop plant architecture.

Research on the Evaluation of Reformation and Revolution System for Universities Based on Neural Network.

Accurately evaluating the working conditions of college revolution and reformation system personnel is currently a hot issue in the field of revolution and reformation system research. Based on the neural network architecture, this paper constructs a model for the system and obtains relevant quality evaluation indicators through simulation experiment analysis. According to the quality evaluation index framework of the revolution and reformation system, the experimental platform selects 7 universities in the region, 6 of which are samples, and 1 (S University) is the research target, and the scores of each index are calculated using MATLAB auxiliary software. The simulation process starts from the characteristics of the actual neural network model; selects 17 evaluation impact indicators, including 3 basic indicators and 14 technical indicators based on historical data; and uses the factor recursive method to improve the neural network and establish an evaluation model. Then use the collected specific data to train a BP neural network with a structure of 21 × 11 × 1, continuously adjust the weights and thresholds of the neural network until the standard error requirements are met, finally verify the scientificity of the evaluation model, and compare the actual output value with the expected output value for comparison. The experimental results show that the model input data redundancy rate is reduced to 0.136, and the network training time delay is reduced to 413 ms, which improves the computing power of the network model of the reformation and revolution system in colleges and universities. The use of neural network data to reduce dimensionality effectively promotes the reformation and revolution system in universities.

The Cancers-Specific Survival of Metastatic Pulmonary Carcinoids and Sites of Distant Metastasis: A Population-Based Study.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths and pulmonary carcinoids (PCs) account for almost 2% of all pulmonary malignancies. However, few published articles have reported prognosis and related factors of pulmonary carcinoid patients. MATERIAL AND METHOD: The Surveillance, Epidemiology, and End Results (SEER) database was used to collect data of patients diagnosed with metastatic PCs from 2010 to 2016. The prognosis and survival of these patients were compared by employing Cox proportional hazards and the Kaplan-Meier survival analysis. RESULTS: A total of 1763 patients were analyzed. The liver (668, 25.6%) was shown to be the most common metastatic site in the isolated organ metastasis cohort, followed by the lung (636, 24.4%), bone (562, 21.6%), and brain (460, 17.6%). Among the patients, the tumor metastasized to a single distant site included the liver, bone, lung, and brain. Cancer-specific survival (CSS) in metastatic PCs is determined by the site of metastasis and the total number of such sites. Pulmonary carcinoid patients with isolated liver metastasis manifested more favorable survival rates in comparison to patients having isolated metastasis in the lung, brain, or bone. The median CSS was 45, 7, 6, 5 months (P = 0.011). The number of distant metastatic sites and the location of distant metastasis were found to be independent risk factors for CSS. For patients with distant isolated metastasis, liver metastasis (P < 0.0001) had better CSS in comparison to those with bone metastasis. When compared to patients whose carcinoids had metastasized to the bones, patients with a brain (P = 0.273) or lung (P = 0.483) metastasis had the same CSS. CONCLUSION: Cancer-specific survival in metastatic PCs depends on the site of metastasis and the total number of such locations. PC patients with isolated liver metastasis manifested more favorable survival in comparison to patients with isolated metastasis in the lung, brain, or bone.

In-situ ZnO template preparation of coal tar pitch-based porous carbon-sheet microsphere for supercapacitor.

Three-dimension (3D) porous carbon-sheet microspheres (PCSMs) are prepared through coating coal tar pitch on basic zinc carbonate microspheres followed by in situ ZnO template carbonization and KOH activation. The as-prepared PCSMs show microsphere morphology composed of petal-like carbon nanosheets, which have large specific area (1359.88-2059.43 m2 g-1) and multiscale pores (mainly micropores and mesopores). As the supercapacitor electrodes, the 3D PCSMs present a good electrochemical performance with a large specific capacitance of 313 F g-1 at 1 A g-1 and high rate capability of 81.9% capacitance retention when increasing the current density up to 50 A g-1 in a three-electrode system. In addition, the energy density can reach up to 18.79 Wh kg-1 at a high power density of 878.4 W kg-1 for PCSMs-0.2a symmetrical supercapcitor in 1 M Na2SO4 electrolyte.

Polysaccharide-Based Lotus Seedpod Surface-Like Porous Microsphere as an Efficient Drug Carrier for Cancer Treatment.

BACKGROUND: This study aimed to evaluate the properties and functions of polysaccharide-based porous microsphere (PPM) for drug delivery, as well as its inhibitory effect on malignant tumors. MATERIALS AND METHODS: PPM was prepared using the inverse emulsion polymerization method. FT-IR measurements were conducted to measure the wavenumber of PPM. Particle size distribution was tested with a particle analyzer, and surface morphologies of PPM were observed using a scanning electron microscope (SEM). Dialysis method, Cell Counting Kit-8 (CCK-8), and cell apoptosis analysis were adopted to evaluate the drug release, cytotoxicity and biocompatibility of mitomycin-C (MMC), respectively. Finally, an in vivo study was performed in C57BL/6 mice to confirm the function of MMC-loaded PPM on tumor growth. RESULTS: FT-IR spectra proved the successful preparation of MMC-loaded PPM. PPM had an average size of 25.90 ± 0.34 µm and then increased to 30.10 ± 0.20 µm after drug loading. Under SEM, the surface morphology was lotus seedpod surface-like, with macropits on the surface and micropores in macropits. Compared with the free MMC group, MMC-loaded PPM exhibited a delayed drug release rate in a pH-dependent manner and higher cell viability. Flow cytometry results showed that the cell apoptosis in the PPM/MMC group was lower than that in the free MMC group. In vivo experiment revealed the inhibitory efficacy of MMC-loaded PPM on malignant tumors. CONCLUSION: In summary, MMC-loaded PPM exhibited favorable surface morphology, sustained drug release ability, nontoxicity and excellent biocompatibility, suggesting that PPM might be a potential drug carrier for tumor treatment.

Anionic Biopolymer Assisted Preparation of MoO2@C Heterostructure Nanoparticles with Oxygen Vacancies for Ambient Electrocatalytic Ammonia Synthesis.

Recently, Mo-based metal catalysts are widely applied in the electrocatalytic nitrogen reduction reaction (NRR) due to the lower binding energy between the Mo atom and N atom. The design of a Mo-based catalyst@carbon heterostructure and the introduction of anion vacancies are effective measures to improve their NRR performance. In this research, the cross-linked Vo-MoO2@C (Vo means oxygen vacancies) heterostructure nanoparticles with rich oxygen vacancies are first synthesized via pectin assisted hydrothermal reaction followed by calcination and treating with NaBH4 solution. Vo-MoO2@C exhibits good electrocatalytic NRR performance with an ammonia yield rate of 9.75 µg h-1 mg-1 at -0.5 V (RHE) and a Faraday efficiency (FE) of 3.24% at -0.3 V (RHE) under ambient conditions.

Biopsy pattern of invasion type to determine the surgical approach in early-stage oral squamous cell carcinoma.

Depth of invasion (DOI) and pattern of invasion (POI) indicate tumor invasiveness of oral squamous cell carcinoma (OSCC). However, preoperative DOI evaluation is challenging, and the correlations between DOI and POI are unknown. We aimed to assess DOI and worst pattern of invasion (WPOI) in early-stage OSCC, and evaluate the preoperative predictive ability of biopsy pattern of invasion (BPOI) for WPOI and DOI. This retrospective study included n = 444 OSCC patients with pT1-2N0M0. The prognostic value of DOI, WPOI, and BPOI and the predictive prognostic option for WPOI and DOI by BPOI were assessed. WPOI (1-3 vs 4-5), but not BPOI, predicted the lowest survival rate and highest DOI. To evaluate the difference between WPOI and BPOI, we conducted a POI type-matching analysis of patients with BPOI1-4 and WPOI1-5. Based on each WPOI type, the false-prediction rates (FPR) of BPOI types 1 (n = 23), 2 (n = 89), 3 (n = 252), and 4 (n = 80) were 52.17%, 52.81%, 36.90%, and 0%, respectively. BPOI4 perfectly predict WPOI 4-5. As the false-predicted BPOI 1-2 was almost WPOI2-3 (79.7%), regardless of the existed FPR, patients with BPOI1-2 have longer survival and lower DOI than those with BPOI 4. However, this phenomenon was not observed in BPOI3, because all false-predicted BPOI3 were WPOI4-5 with a high DOI. We provide an alternative predictive prognostic option for WPOI and DOI by evaluating BPOI during OSCC surgical planning, with the recommendation of conservative treatment in patients with BPOI 1-2.

Survival nomogram for patients with metastatic siewert type II adenocarcinoma of the esophagogastric junction: a population-based study.

BACKGROUND: The aim of this study was to construct a nomogram to predict the survival of patients with metastatic Siewert Type II adenocarcinomas of the esophagogastric junction (AEG). METHODS: Patients were identified using the Surveillance, Epidemiology, and End Results (SEER) database. Cox regression analysis was performed to assess the prognostic factors. A nomogram comprising independent prognostic factors was established and evaluated using C-indexes, calibration curves, and decision curve analyses. RESULTS: In total 1616 eligible patients were enrolled. Race, age, bone metastasis, liver metastasis, lung metastasis, other metastasis sites, and distant lymph nodes metastasis were independent prognostic factors and were integrated to construct the nomogram. The nomogram had a C-index of 0.590 (95% CI: 0.569-0.611) in the training cohort and 0.569 (95% CI: 0.532-0.606) in the validation cohort. The calibration plots for the probabilities of 6-month and 1-year overall survival demonstrated there was an optimum between nomogram prediction and actual observation. CONCLUSION: We developed and validated a nomogram to predict individual prognosis for patients with metastatic Siewert Type II AEG, and the risk stratification system based on the nomogram could effectively stratify the patients into two risk subgroups, which can help clinicians accurately predict mortality risk and recommend personalized treatment modalities.

Sites of distant metastases and the cancer-specific survival of metastatic Siewert type II esophagogastric junction adenocarcinoma: a population-based study.

BACKGROUND: The effect of distant metastasis on prognosis in patients with Siewert type II adenocarcinoma of the esophagogastric junction (AEG) remains elusive. METHODS: Patients diagnosed as metastatic Siewert type II AEG were identified using the Surveillance, Epidemiology, and End Results (SEER) database. Kaplan-Meier survival analysis and a Cox proportional hazards analysis were performed to assess the effect of distant metastases sites. RESULTS: We analyzed 1616 eligible patients. Liver was the most frequent metastatic site. For patients with isolated distant metastasis, the median survival time was 8, 7, 8, 10, and 11 months for patients with liver, bone, brain, lung, and distant lymph nodal metastasis, respectively (p = 0.011). The number of metastatic sites and the site of distant metastasis were independent prognostic factors for cancer-specific survival (CSS). In patients with isolated distant metastasis, using bone metastasis as reference, lung (p = 0.011) or distant lymph node metastasis (p = 0.030) was associated with better CSS, while patients with liver (p = 0.051) or brain (p = 0.488) metastasis had similar CSS compared to patients with bone metastasis. CONCLUSION: CSS in metastatic Siewert type II AEG is dependent on the metastatic site and the number of metastatic sites.

Mechanism of epithelial­mesenchymal transition inhibited by miR­203 in non­small cell lung cancer.

The aim of the present study was to investigate whether miR­203 can inhibit transforming growth factor­ß (TGF­ß)­induced epithelial­mesenchymal transition (EMT), and the migration and invasion ability of non­small cell lung cancer (NSCLC) cells by targeting SMAD3. In the present study, the expression levels of miR­203, SMAD3 mRNA and protein in NSCLC tissues were examined, as well as their corresponding paracancerous samples. The miR­203 mimics and miR­203 inhibitor were transfected into the H226 cell line. RT­qPCR was used to assess the expression levels of E­cadherin, Snail, N­cadherin and vimentin mRNA, and western blotting was performed to detect the expression levels of p­SMAD2, SMAD2, p­SMAD3, SMAD3 and SMAD4. The cell migration and invasion abilities were detected by Transwell assays. The target site of SMAD3 was predicted by the combined action between miR­203 and dual luciferase. The results revealed that the RNA levels of miR­203, compared with paracancerous tissues, were decreased in NSCLC tissues, while SMAD3 mRNA and protein levels were upregulated, and miR­203 inhibited SMAD3 expression. Induction of TGF­ß led to decreased E­cadherin mRNA levels, upregulation of Snail, N­cadherin and vimentin mRNA levels (P<0.05), and significant increase in cell migration and invasion, whereas transfection of miR­203 mimics reversed the aforementioned results (P<0.05). Conversely, miR­203 inhibitor could further aggravate the aforementioned results (P<0.05). Western blot results revealed that transfection of miR­203 mimics significantly reduced the protein expression of SMAD3 and p­SMAD3 (P<0.05). Furthermore, the results of the Dual­Luciferase assay revealed that miR­203 inhibited SMAD3 expression by interacting with specific regions of its 3'­UTR. Overall, a novel mechanism is revealed, in which, miR­203 can inhibit SMAD3 by interacting with specific regions of the 3'­UTR of SMAD3, thereby restraining TGF­ß­induced EMT progression and migration and invasion of NSCLC cells.

Clinical features and prognosis of primary tracheal small cell carcinoma: a population-based analysis.

BACKGROUND: Primary tracheal small cell carcinoma (SCC) is an uncommon malignancy; therefore, its clinical features and prognosis are still unclear. METHODS: We used the population-based Surveillance, Epidemiology, and End Results (SEER) database to elucidate the clinical features and prognosis of primary tracheal SCC. The clinical features were assessed by using the chi-square test. Overall survival (OS) was computed using the Kaplan-Meier method, and a Cox proportional hazards analysis was performed to evaluate the prognostic factors. RESULTS: From 1973 to 2015, 1,392 primary tracheal tumor cases were reported in the SEER database, 75 (5.4%) of which were SCC. Age, sex, race, extent of disease, lymph node involvement, surgery and radiation treatment were similar between patients with SCC and those with squamous cell carcinoma (SQC), but patients with SCC were more likely to receive chemotherapy (65.8% vs. 28.2%, respectively; P<0.001). The 1-, 3- and 5-year OS rates of patients with SCC were 37.8%, 12.4% and 7.1%, respectively, and the median survival duration was 10.0 months, which was much worse than that observed for patients with other histopathological types of tracheal cancer. Among patients aged 60-100 years and those with regional lymph node involvement, the OS for patients with SQC was superior to that for patients with SCC (P=0.034 and P=0.016, respectively). According to the multivariate analysis, age and lymph node involvement are independent prognostic factors of SCC. CONCLUSION: Primary tracheal SCC is a rare carcinoma with a poor prognosis. Age and lymph node involvement are independent prognostic factors of SCC.

Milk-exosome based pH/light sensitive drug system to enhance anticancer activity against oral squamous cell carcinoma.

A multimodal drug delivery system targeting the tumor microenvironment is an inspiring method for treating cancer tissues, including oral squamous cell carcinomas (OSCC). Such approaches require an efficient and safe drug carrier. Bovine milk derived exosomes are ideal because the source is adequate and have advantages of both synthetic and cell-mediated nano carriers. In the present study, we developed a pH/light sensitive drug system based on milk-exosomes for OSCC therapy. It was called exosome-doxorubicin-anthracene endoperoxide derivative (Exo@Dox-EPT1, NPs). Milk-exosomes were conjugated to doxorubicin (Dox) by a pH-cleavable bond, which can rapture under an acidic microenvironment. Besides, endoperoxides and chlorin e6 (Ce6) were also loaded and the endoperoxides undergo thermal cycloreversion and release singlet oxygen to kill cancer cells. We have also investigated the body distribution, antitumor effects, and biocompatibility of the nanoparticles. The new milk-exosome-based drug delivery system showed controlled drug-release, biocompatibility and, proved to be effective in treating OSCC.

NALP3 orchestrates cellular bioenergetics to facilitate non-small cell lung cancer cell growth.

AIMS: Previous work has reported the closely correlation between inflammation and carcinogenesis, while the role of NALP3, the key component of inflammasome activation in NSCLC remains elusive. This study was to unravel the mechanism of NALP3 on modulating NSCLC cancer cell growth. METHODS: IHC and immuno-blot were performed to analyze expression of NALP3 and indicated molecules. CCK-8 and xenograft nude mice assay were used to evaluate cell growth in vitro and in vivo. Bioenergetics assay was performed to measure OXPHOS and aerobic glycolysis. siRNA and shRNA were constructed to knockdown endogenous NALP3 and DNMT1. Co-immunoprecipitation was applied to confirm the interaction between NALP3 and DMAP1. BioProfile FLEX analyzer and Lactate Reagent Kit were used to measure relative level glucose uptake and lactate production. KEY FINDINGS: We reported NALP3 were up-regulated in NSCLC tumor tissues. NALP3 depletion suppressed cancer cell growth in vitro and in vivo. Moreover, data showed depletion of NALP3 promoted cell bioenergetics switch from aerobic glycolysis to OXPHOS. Additionally, we found NALP3 interacted with DMAP1 and alteration of NALP3 increased DNMT1 level. Subsequently, we clarified depletion of DNMT1 significantly suppressed NSCLC cell growth and orchestrated cellular metabolism which was similar to the effects of NALP3 knockdown. Finally, our data showed high NALP3 was associated with poor outcomes, and correlated with TNM stage and differentiation. SIGNIFICANCE: Current study elucidated NALP3 could promote metabolic reprogramming to regulate NSCLC cell growth and suggested that NALP3 may be considered as a novel biomarker and therapeutic target for NSCLC patients.

Gephyrin suppresses lung squamous cell carcinoma development by reducing mTOR pathway activation.

Background: The mTOR pathway is altered in a multitude of cancers, including lung cancer; however, abnormal activation in this pathway is less common in lung adenocarcinoma (LUAD) than in lung squamous cell carcinoma (LUSC). Gephyrin is a highly conserved and widely expressed ancient protein in vertebrate tissues. Its role and molecular mechanism in lung cancer development are largely unknown. Method: We analyzed the expression profile of gephyrin and overall survival rates in LUAD and LUSC. The LUSC cells (H520 and SK-MES-1) were transfected with pLV-gephyrin to establish gephyrin stable overexpression cell lines. Real-time quantitative PCR and Western blot were performed to detect the mRNA and protein levels. The cell growth and cell cycle were detected by the MTT assay and flow cytometry. Finally, a xenograft tumor model was established to determine cell tumorigenesis in vivo. Results: Our results show that gephyrin was reduced in LUAD and LUSC, and its low expression in LUSC patients indicated poor prognosis. Gephyrin overexpression suppressed LUSC cell proliferation, arrested cell cycle progression, and decreased the expression of cell-cycle related proteins such as cyclin D1, cyclin-dependent kinase-2 (CDK2), and proliferation-related protein proliferating cell nuclear antigen (PCNA). Conversely, knockdown of gephyrin promoted LUSC cell growth. Moreover, gephyrin reduced mTOR pathway activation to inhibit cyclin D1 and CDK2 translation. Mechanistically, gephyrin suppressed mTOR pathway activation by promoting mTOR degradation. Furthermore, gephyrin overexpression suppressed LUSC tumorigenesis. Conclusion: Gephyrin suppressed LUSC development by reducing mTOR pathway activation, implicating gephyrin as a potential molecular target for LUSC management.

MicroRNA-488-3p inhibits proliferation and induces apoptosis by targeting ZBTB2 in esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is the eighth most prevalent cancer and the sixth leading cause for cancer-associated mortality. MicroRNAs (miRNAs) are increasingly reported to exert important regulatory functions in human cancers by regulating certain gene expression. miR-488-3p has been identified to be a tumor suppressor in multiple cancers, but its role in ESCC is yet to be investigated. The present study aimed to uncover the biological role and modulatory mechanism of miR-488-3p in ESCC. We first revealed the downregulation of miR-488-3p in ESCC tissues and cell lines. Gain-of-function assays confirmed that miR-488-3p overexpression abrogated proliferation and accelerated apoptosis. Mechanistically, we identified via bioinformatics tool and confirmed that zinc finger and BTB domain containing 2 (ZBTB2) was a target for miR-488-3p. Moreover, miR-488-3p activated the p53 pathway through suppressing ZBTB2. Finally, rescue assays proved that ZBTB2 was involved in the regulation of miR-488-3p on proliferation and apoptosis in ESCC. Additionally, we verified that miR-488-3p had alternate targets in ESCC by confirming the involvement of protein kinase, DNA-activated, catalytic subunit (PRKDC), a known target for miR-488-3p, in miR-488-3p-mediated regulation on ESCC. In sum, this study revealed that miR-488-3p inhibited proliferation and induced apoptosis by targeting ZBTB2 and activating p53 pathway in esophageal squamous cell carcinoma, providing a novel biological target for ESCC.

Integrated analysis reveals five potential ceRNA biomarkers in human lung adenocarcinoma.

BACKGROUND: Competing endogenous RNAs (ceRNAs) are a newly identified type of regulatory RNA. Accumulating evidence suggests that ceRNAs play an important role in the pathogenesis of diseases such as cancer. Thus, ceRNA dysregulation may represent an important molecular mechanism underlying cancer progression and poor prognosis. In this study, we aimed to identify ceRNAs that may serve as potential biomarkers for early diagnosis of lung adenocarcinoma (LUAD). METHODS: We performed differential gene expression analysis on TCGA-LUAD datasets to identify differentially expressed (DE) mRNAs, lncRNAs, and miRNAs at different tumor stages. Based on the ceRNA hypothesis and considering the synergistic or feedback regulation of ceRNAs, a lncRNA-miRNA-mRNA network was constructed. Functional analysis was performed using gene ontology term and KEGG pathway enrichment analysis and KOBAS 2.0 software. Transcription factor (TF) analysis was carried out to identify direct targets of the TFs associated with LUAD prognosis. Identified DE genes were validated using gene expression omnibus (GEO) datasets. RESULTS: Based on analysis of TCGA-LUAD datasets, we obtained 2,610 DE mRNAs, 915 lncRNAs, and 125 miRNAs that were common to different tumor stages (|log2(Fold change)| ≥ 1, false discovery rate < 0.01), respectively. Functional analysis showed that the aberrantly expressed mRNAs were closely related to tumor development. Survival analyses of the constructed ceRNA network modules demonstrated that five of them exhibit prognostic significance. The five ceRNA interaction modules contained one lncRNA (FENDRR), three mRNAs (EPAS1, FOXF1, and EDNRB), and four miRNAs (hsa-miR-148a, hsa-miR-195, hsa-miR-196b, and hsa-miR-301b). The aberrant expression of one lncRNA and three mRNAs was verified in the LUAD GEO dataset. Transcription factor analysis demonstrated that EPAS1 directly targeted 13 DE mRNAs. CONCLUSION: Our observations indicate that lncRNA-related ceRNAs and TFs play an important role in LUAD. The present study provides novel insights into the molecular mechanisms underlying LUAD pathogenesis. Furthermore, our study facilitates the identification of potential biomarkers for the early diagnosis and prognosis of LUAD and therapeutic targets for its treatment.