Silencing of lncRNA LOC105376794 promotes migration, invasion, and Gefitinib resistance of lung adenocarcinoma cells with EGFR 19Del mutation by ATF4/CHOP axis and ERK phosphorylation.

Epidermal growth factor receptor (EGFR) gene exon 19 in-frame deletion (19del) and exon 21 L858R point mutation (21L858R mutation) are prevalent mutations in lung adenocarcinoma. Lung adenocarcinoma patients with 19del presented with a better prognosis than the 21L858R mutation under the same epidermal growth factor receptor tyrosine kinase inhibitor treatment. Our study aimed to uncover the expression of long non-coding RNA LOC105376794 between 19del and 21L858R mutation, and explore the mechanism that regulates cells' biological behavior and gefitinib sensitivity in lung adenocarcinoma cells with 19del. Transcriptome sequencing was conducted to identify differentially expressed lncRNAs between EGFR 19del and 21L858R mutation in serum through the DNBSEQ Platform. Protein-protein interaction network and Kyoto Encyclopedia of Genes and Genomes pathway were conducted to analyze the relationship between lncRNAs and mRNAs through STRING and Dr. TOM. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to measure the expression of lncRNA LOC105376794 in serum and cells. Loss-of-function experiments were used to validate the biological function and gefitinib sensitivity of LOC105376794 in lung adenocarcinoma cells. Protein levels were detected by western blotting. Through transcriptome resequencing and RT-qPCR, we found the expression levels of LOC105376794 in serum were increased in the 19del group compared with the 21L858R mutation group. Inhibition of LOC105376794 promoted proliferation, migration and invasion, and reduced apoptosis of HCC827 and PC-9 cells. The low expression of LOC105376794 reduced gefitinib sensitivity in PC-9 cells. Mechanistically, we found that the knockdown of LOC105376794 suppressed activating transcription factor 4 (ATF4)/C/EBP homologous protein (CHOP) signaling pathway and facilitated the expression of extracellular signal-regulated kinase 1/2 (ERK) phosphorylation. LOC105376794 altered cell biological behavior and gefitinib sensitivity of lung adenocarcinoma cells with 19del through the ATF4/CHOP signaling pathway and the expression of ERK phosphorylation. The results further illustrated the fact that lung adenocarcinoma patients with 19del presented with a more favorable clinical outcome and provided a theoretical basis for treatment strategy for lung adenocarcinoma patients with 19del.

Secretin-dependent signals in the ventromedial hypothalamus regulate energy metabolism and bone homeostasis in mice.

Secretin, though originally discovered as a gut-derived hormone, is recently found to be abundantly expressed in the ventromedial hypothalamus, from which the central neural system controls satiety, energy metabolism, and bone homeostasis. However, the functional significance of secretin in the ventromedial hypothalamus remains unclear. Here we show that the loss of ventromedial hypothalamus-derived secretin leads to osteopenia in male and female mice, which is primarily induced by diminished cAMP response element-binding protein phosphorylation and upregulation in peripheral sympathetic activity. Moreover, the ventromedial hypothalamus-secretin inhibition also contributes to hyperphagia, dysregulated lipogenesis, and impaired thermogenesis, resulting in obesity in male and female mice. Conversely, overexpression of secretin in the ventromedial hypothalamus promotes bone mass accrual in mice of both sexes. Collectively, our findings identify an unappreciated secretin signaling in the central neural system for the regulation of energy and bone metabolism, which may serve as a new target for the clinical management of obesity and osteoporosis.

The Influence of Storage in Saline or Irradiation by Ultraviolet on Surface Hydrophilicity of Implant and Osseointegration: An Experimental Study in Rabbits.

Storage in aqueous solution or ultraviolet (UV) irradiation can retain or regain the hydrophilicity of titanium implant surface. In this study, 3 types of commercial titanium implants were used: ZBL (ZDI Bone Level), CEL (C-tech Esthetic Line), and modSLA (Straumann SLActive). ZBL and CEL implants were treated with UV irradiation for 4 hours. Surface characterization of the 4 groups (ZBL, ZBL-UV, CEL-UV, and modSLA) was evaluated by scanning electron microscopy and contact angle measurements. The in vivo bone response was evaluated by removal torque (RTQ) tests and histomorphometric analysis at 3, 6, and 12 weeks postimplantation. A total of 144 implants and 36 rabbits were used for experiments according to a previously established randomization sequence. The ZBL-UV, CEL-UV, and modSLA groups were hydrophilic, and nanostructures were observed on the modSLA implant surface. ModSLA achieved better RTQ value than ZBL at 12 weeks (P < .05). For histomorphometric analysis, ZBL-UV and CEL-UV implants showed higher bone area values in the cancellous bone zone at 6 weeks than did modSLA and ZBL implants (P < .05). In the cortical bone zone, all groups showed comparable bone-to-implant contact at all healing time points (P > .05). Both storage in saline and UV irradiation could retain or provoke hydrophilic surfaces and improve osseointegration. Compared with storage in saline, UV irradiation displayed slight advantages in promoting new bone formation in cancellous bone zone at an early stage.

Identification of Key Pathways and Genes Downstream of Insulin-Like Growth Factor 1 in Thyroid Carcinoma.

Introduction: Thyroid carcinoma (THCA) is the most common endocrine tumor worldwide. Insulin-like growth factor 1 (IGF1) is a polypeptide hormone with a high degree of structural similarity to human proinsulin. Materials and Methods: We used online data to investigate the expression pattern of IGF1 in THCA. We also identified gene signatures and pathways downstream of IGF1 to elucidate potential mechanisms. Further analysis identified critical hub genes and pathways using the STRING database and the DAVID online tool. We analyzed gene expression and the prognostic quality of hub genes using the GEPIA online tool and the Human Protein Atlas website. Results: IGF1 expression was found to be downregulated in THCA tissues, both those in the TCGA database and in tissues recovered during surgery at our hospital. All samples from the THCA-TCGA dataset were divided into IGF1 high- and low-expression groups. We identified 1014 differentially expressed genes (DEGs) between the two groups. Functional enrichment analysis showed that DEGs were mainly concentrated in immune response, cell adhesion, and cancer-related pathways. We then identified the top hub genes and performed a prognostic analysis to identify the most critical genes. The expression levels of FN1, MMP9, and CD40LG correlated with disease prognosis. Finally, we confirmed the expression levels of these genes using the HPA website. Conclusion: Our results identified potential key genes and pathways downstream of IGF1 in THCA, providing insight into the mechanisms underlying the development of THCA.

CircLATS2 Regulates miR-520a-3p/E2F7/p-VEGFR2 Signaling Pathway to Promote Hepatocellular Carcinoma Progression and Angiogenesis.

Objective: To investigate the effect of circLATS2 on the progression and angiogenesis of hepatocellular carcinoma and its molecular mechanism. Methods: The expression of circLATS2 in hepatocellular carcinoma was detected by qRT-PCR. The StarBase database was used to predict the potential miRNA, and the combination of the above was cytological verified by luciferase reporter gene assay and RNA pull down. The potential target genes of miRNA were predicted by TargetScan, verified by the above experiments, and the influence of circLATS2 on its expression was determined. The biological function of circLATS2 was investigated by in vitro and in vivo experiments. The effects of miRNA and target genes on the malignant behavior of HCC cells were determined by the reverse experiment. Results: circLATS2 was highly expressed in HCC and was positively correlated with tumor size and tumor stage. miR-520a-3p was sponged by circLATS2 and was low expressed in HCC tissues. As the target gene of miR-520a-3p, the expression level of E2F7 is affected by circLATS2. In vitro experiments showed that circLATS2 knockdown inhibited the proliferation, clone formation, migration, and invasion ability of hepatocellular carcinoma cells. In vivo knockdown of circLATS2 inhibits the proliferation of HCC cells, while overexpression of circLATS2 promotes the proliferation of HCC cells. Overexpression of miR-520a-3p and E2F7 knockdown reversed the role of circLATS2 in promoting malignant behavior of HCC cells and affected phosphorylation of VEGFR2. Conclusion: CircLATS2 promotes the progression of HCC by regulating miR-520a-3p/E2F7/P-VEGFR2 signaling pathway.

Radiographic assessment of deproteinized bovine bone mineral (DBBM) and collagen-stabilized DBBM for transalveolar sinus floor elevation: A 2-year retrospective cohort study.

BACKGROUND: Theoretically, collagen-stabilized deproteinized bovine bone mineral (DBBM-C) has better operability compared with DBBM. DBBM-C avoids dispersing during the transalveolar sinus floor elevation (TSFE) because of its block shape. PURPOSE: To evaluate radiological changes of using DBBM-C in TSFE. MATERIALS AND METHODS: Patients who received TSFE using DBBM (Bio-Oss®) or DBBM-C (Bio-Oss® collagen) with simultaneous implantation were recruited. Graft bone height apically (aGH), endo-sinus bone gain (ESBG), and crest bone level (CBL) were assessed through panoramic radiograph and cone beam computed tomography (CBCT). RESULTS: A total of 138 patients (138 implants) were retrospectively enrolled. After 2 years of implantation, the incidence of postoperative complications was 4.2% (95% CI: 0.9%-11.7%) and 4.5% (95% CI: 0.9%-12.7%) for DBBM and DBBM-C groups, respectively. Measured in panoramic radiograph, ΔaGH of DBBM-C (1.8 mm, SD: 1.4, 95% CI: 1.2-2.4, P = 0.044) group was significantly higher than that of DBBM (1.2 mm, SD: 1.4, 95% CI: 0.7-1.7) after 24 months. No significant differences for ΔCBL were noted during the entire observation period. Measured through CBCT, ESBG was 5.0 (SD: 1.8, 95% CI: 4.1-5.8) mm in DBBM group and 4.6 (SD: 1.6, 95% CI: 3.9-5.3) mm in DBBM-C group 24 months after implantation. The aGH value of DBBM-C group was significantly higher compared with DBBM in CBCT (OR = 1.4, 95% CI: 1.1-1.9, P = 0.020). CONCLUSIONS: DBBM-C could achieve similar bone generation as DBBM in TSFE. Both materials could maintain aGH, ESBG, and CBL relatively stable 2 years after implantation.

The Association of ERCC1 and ERCC5 Polymorphisms with Lung Cancer Risk in Han Chinese.

Background: Polymorphisms in DNA damage repair genes are important determinants for cancer susceptibility, clinical phenotype diversity, and therapy. However, their relationship with lung cancer remains unclear. This study aimed to investigate the role of DNA damage repair gene polymorphisms in the risk of lung cancer. Methods: The matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectroscopy-based genotyping system was used to genotype 601 individuals (200 lung cancer patients and 401 age- and sex-matched healthy controls) for polymorphisms in excision repair cross-complementing group 1 (ERCC1) and ERCC5 genes. Results: The ERCC5 rs4771436 GG genotype, recessive model (GG vs. GT+TT), and the ERCC5 rs1047768 recessive model (CC vs. CT+TT) were associated with significantly increased risks of lung cancer (P=0.029, P=0.014, and P=0.044, respectively), especially in men and individuals aged 60 years or younger. Conclusion: ERCC5 rs4771436 and rs1047768 genotypes were associated with an increased risk of lung cancer, suggesting that polymorphisms in DNA repair genes are significantly related to the risk of lung cancer, and play an important role in the occurrence of lung cancer.

Identification of PGC-related ncRNAs and their relationship with the clinicopathological features of Gastric Cancer.

Pepsinogen C (PGC) is considered to be the final product of mature differentiated gastric mucosa. The expression level of PGC in gastric mucosa is clearly decreased upon the development of gastric cancer (GC). However, the mechanism behind PGC's down-regulation remains unclear and needs to be clarified. This study aimed to identify PGC-related ncRNAs with the potential to be PGC post-transcriptional regulators and to further explore the association between these ncRNAs and the clinicopathological parameters of GC. Bioinformatic software was used to predict miRNAs binding specifically to PGC and circRNAs binding specifically to these candidate miRNAs. Dual-luciferase reporter assay was performed to validate the completely complementary pairing of PGC and PGC-related ncRNAs. qRT-PCR was applied to determine the expression levels of PGC and PGC-related ncRNAs in GC tissue. hsa-let-7c was predicted to bind to the PGC gene, which was confirmed by dual-luciferase reporter assay. hsa_circ_0001483 and hsa_circ_0001324 were identified to bind to hsa-let-7c by bioinformatic analysis and dual-luciferase reporter assay. In addition, the hsa_circ_0001483/hsa_circ_0001324 -hsa-let-7c-PGC axis was confirmed in tissue by qRT-PCR. The expression level of hsa_circ_0001483 was correlated with peritumoral inflammatory cell infiltration and lymphatic metastasis. hsa_circ_0001483, hsa_circ_0001324, and let-7c were newly identified and validated as PGC-related ncRNAs and showed associations with the clinicopathological features of GC. The hsa_circ_0001483/hsa_circ_0001324-hsa-let-7c-PGC axis in GC may account for the down-regulation of PGC in GC tissue.

Biomechanical and Morphometric Evaluation of a Strontium-Incorporated Implant and Four Commercially Available Implants: A Study in Rabbits.

PURPOSE: Strontium has shown a positive effect on osseointegration in experiments. This study compared surface characterization and osseointegration of a strontium-incorporated implant with four commercial implants with different surface treatments. MATERIALS AND METHODS: A strontium-oxide layer was created by hydrothermal treatment on the surface of the implant (SLA-Sr). Surface characterizations were observed using a scanning electron microscope, three-dimensional (3D) optical microscope, and x-ray energy-dispersive spectrometry. Implants of different surface treatments including resorbable blasting media (RBM), sandblasting with large grit and acid etching (SLA-1, SLA-2), sandblasting and thermal acid etching (STA), and SLA-Sr were implanted into the proximal tibiae and femoral condyles of rabbits. Biologic effects were evaluated by removal torque testing and histomorphometric analysis after 3, 6, and 12 weeks of implantation. RESULTS: Nanostructures were observed on the surface of SLA-Sr and STA. Calcium (Ca) was detected on the surface of RBM. Sr was detected on the surface of SLA-Sr. SLA-1 and STA had greater surface roughness than SLA-2, SLA-Sr, and RBM (P < .05). In vivo, SLA-Sr achieved better removal torque value (RTV) than that of RBM and SLA-2 at 3 weeks (P < .05), as well as increased bone area ratio (BA%) in cortical bone compared with RBM at 3 weeks (P < .05). STA showed higher bone-to-implant contact ratio (BIC%) in cortical bone than RBM at 3 and 6 weeks (P < .05). Compared with RBM, SLA-1 had better RTV at 6 weeks and higher BIC% in cortical bone at 12 weeks (P < .05). CONCLUSION: In vivo, compared with SLA-2 and RBM, the implant with the strontium-oxide layer displayed slight advantages in new bone formation and osseointegration in the early healing stage. In the later osseointegration stage, the results of SLA-Sr were comparable with other implants.

Cell-based immunotherapy with cytokine-induced killer (CIK) cells: From preparation and testing to clinical application.

Cell-based immunotherapy holds promise in the quest for the treatment of cancer, having potential synergy with surgery, chemotherapy and radiotherapy. As a novel approach for adoptive cell-based immunotherapy, cytokine-induced killer (CIK) cells have moved from the 'bench to bedside'. CIK cells are a heterogeneous subset of ex-vitro expanded, polyclonal T-effector cells with both natural killer (NK) and T-cell properties, which present potent non-major histocompatibility complex-restricted cytotoxicity against a variety of tumor target cells. Initial clinical studies on CIK cell therapy have provided encouraging results and revealed synergistic antitumor effects when combined with standard therapeutic procedures. At the same time, issues such as inadequate quality control and quantity of CIK cells as well as exaggerated propaganda were continuously emerging. Thus, the Ministry of Health in China stopped CIK cell therapy in May 2016, which was a major setback for the innovation of CIK cell-based immunotherapy. Thus, it is very important to modify technical criteria to develop a standardized operation procedure (SOP) and standardized system for evaluating antitumor efficacy in a safe way.

Association of Polymorphisms in three pri-miRNAs that Target Pepsinogen C with the Risk and Prognosis of Gastric Cancer.

We aimed to explore the associations of polymorphisms in three microRNAs (miRNAs) (let-7e rs8111742, miR-365b rs121224 and miR-4795 rs1002765) that target PGC with the risk and prognosis of gastric cancer/atrophic gastritis. Sequenom's MassArray was used to genotype the miRNA polymorphisms in 724 gastric cancer cases, 862 atrophic gastritis cases and 862 controls in a Chinese population. We found that let-7e rs8111742 and miR-4795 rs1002765 were associated with the risk of gastric cancer in the H. pylori-positive subgroup. MiR-365b rs121224 was associated with the risk of intestinal-type gastric cancer in the alcohol consumption subgroup. Intestinal-type gastric cancer patients at Borrmann stages III-IV who carry the miR-365b rs121224 GG genotype had better prognosis compared with those who carry the CG or CC genotypes. MiR-365b rs121224 was associated with Lauren typing and TNM staging, in which the distribution of GG genotype carriers in intestinal-type gastric cancer and the TNM stage I-II subgroup was higher than that of CG or CC genotypes, which contrasted with the distribution in diffuse-type gastric cancer or TNM III-IV groups. These findings suggested that the polymorphisms in these miRNAs might be biomarkers for gastric cancer risk and prognosis, especially for populations infected with Helicobacter pylori or who consume alcohol.

SNP-SNP interactions of three new pri-miRNAs with the target gene PGC and multidimensional analysis of H. pylori in the gastric cancer/atrophic gastritis risk in a Chinese population.

Gastric cancer (GC) is a multistep complex disease involving multiple genes, and gene-gene interactions have a greater effect than a single gene in determining cancer susceptibility. This study aimed to explore the interaction of the let-7e rs8111742, miR-365b rs121224, and miR-4795 rs1002765 single nucleotide polymorphisms (SNPs) with SNPs of the predicted target gene PGC and Helicobacter pylori status in GC and atrophic gastritis (AG) risk. Three miRNA SNPs and seven PGC SNPs were detected in 2448 cases using the Sequenom MassArray platform. Two pairwise combinations of miRNA and PGC SNPs were associated with increased AG risk (let-7e rs8111742 - PGC rs6458238 and miR-4795 rs1002765 - PGC rs9471643). Singly, miR-365b rs121224 and PGC rs6912200 had no effect individually but in combination they demonstrated an epistatic interaction associated with AG risk. Similarly, let-7e rs8111742 and miR-4795 rs1002765 SNPs interacted with H. pylori infection to increase GC risk (rs8111742: Pinteraction = 0.024; rs1002765: Pinteraction = 0.031, respectively). A three-dimensional interaction analysis found miR-4795 rs1002765, PGC rs9471643, and H. pylori infection positively interacted to increase AG risk (Pinteraction = 0.027). Also, let-7e rs8111742, PGC rs6458238, and H. pylori infection positively interacted to increase GC risk (Pinteraction = 0.036). Furthermore, both of these three-dimensional interactions had a dosage-effect correspondence (Ptrend < 0.001) and were verified by MDR. In conclusion, the miRNAs SNPs (let-7e rs8111742 and miR-4795 rs1002765) might have more superior efficiency when combined with PGC SNPs and/or H. pylori for GC or AG risk than a single SNP on its own.

Correlation between Vascular Endothelial Growth Factor and Somatostatin Receptor with Progression and Prognosis in Gastric Cancer.

BACKGROUND/AIMS: To investigate the expression of vascular endothelial growth factor (VEGF) and somatostatin receptor (SSTR) and their clinicopathological and prognostic value in gastric cancer (GC). METHODOLOGY: The expression of VEGF and SSTR in 107 cases of GC tissue and 30 cases of gastric mucosa were detected by immunohistochemistry. Clinicopathological and prognostic association of VEGF and SSTR in GC was analyzed RESULTS: The expression of VEGF in GC (70.1%) was significantly higher than that in gastric mucosa (20.0%) The expression of SSTR in GC (62.6%) was significantly lower than that in normal tissues (93.3%). VEGF and SSTR expression were both associated with histological differentiation, depth of invasion, TNM stage, and lymph node metastasis (P < 0.05). The negative expression of VEGF or the positive expression of SSTR was correlated with better overall survival of GC patients. The Cox analysis demonstrated that the expression of VEGF and SSTR, pathological type, TNM stage, and lymph node metastasis were the independent predictors for overall survival in GC (P = 0.005, P = 0.006, P = 0.003, P = 0.002, and P = 0.001, respectively). CONCLUSIONS: The expression of VEGF and SSTR were associated with progression and prognosis of GC.

DNA purification on a lab-on-valve system incorporating a renewable microcolumn with in situ monitoring by laser-induced fluorescence.

Bead injection in a lab-on-valve (LOV) system was adopted for DNA purification via micro solid-phase extraction (SPE) with a renewable silica microcolumn packed in a channel of the LOV unit. The complex matrix components in human whole blood, including proteins, were well eliminated by choosing properly the sample loading and elution media. The DNA purification process was monitored on-line by using laser-induced fluorescence in a demountable side part of the LOV unit incorporating optical fibers. The practical applicability of the entire system was demonstrated by separation/purification of lambda-DNA in a simulated matrix and human blood genetic DNA by performing SPE, in situ monitoring of the purified products, and postcolumn PCR amplification. When DNAs in a simulated matrix (10.0 ng microl-1 lambda-DNA, 50 ng microl-1 bovine serum albumin, 1.0% Triton X-100) were processed in the present system and laser-induced fluorescence was monitored at 610 nm, an overall extraction/collection efficiency of 70% was achieved by employing identical sample loading and an elution flow rate of 0.5 microl s-1, along with a precision of 3.8% relative standard deviation. DNA separation and purification from human whole-blood samples were performed under similar conditions.