Investigation of Tumor Suppressing Function of CACNA2D3 in Esophageal Squamous Cell Carcinoma.

Background: Deletion of 3p is one of the most frequent genetic alterations in esophageal squamous cell carcinoma (ESCC), suggesting the existence of one or more tumor suppressor genes (TSGs) within these regions. In this study, one TSG, CACNA2D3 at 3p21.1, was characterized. Methods: Expression of CACNA2D3 in ESCCs was tested by quantitative real-time PCR and tissue microarray. The mechanism of CACNA2D3 downregulation was investigated by methylation-specific polymerase chain reaction (MS-PCR). The tumor suppressive function of CACNA2D3 was characterized by both in vitro and in vivo tumorigenic assays, cell migration and invasion assays. Results: CACNA2D3 was frequently downregulated in ESCCs (24/48, 50%), which was significantly associated with promoter methylation and allele loss (P<0.05). Tissue microarray result showed that downregulation of CACNA2D3 was detected in (127/224, 56.7%) ESCCs, which was significantly associated with lymph node metastasis (P = 0.01), TNM staging (P = 0.003) and poor outcome of ESCC patients (P<0.05). Functional studies demonstrated that CACNA2D3 could inhibit tumorigenicity, cell motility and induce apoptosis. Mechanism study found that CACNA2D3 could arrest cell cycle at G1/S checkpoint by increasing expressions of p21 and p53 and decreasing expression of CDK2. In addition, CACNA2D3 could upregulate intracellular free cytosolic Ca2+ and subsequently induce apoptosis. Conclusion: CACNA2D3 is a novel TSG responsible to the 3p21 deletion event and plays a critical suppressing role in the development and progression of ESCC. © 2013 Li et al.link_to_OA_fulltex

S100A4 Knockout Sensitizes Anaplastic Thyroid Carcinoma Cells Harboring BRAFV600E/Mt to Vemurafenib

Background/Aims: Anaplastic thyroid cancer (ATC), with 25% BRAFV600E mutation, is one of the most lethal human malignancies that currently has no effective therapy. Vemurafenib, a BRAFV600E inhibitor, has shown promise in clinical trials, including ATC patients, but is being hampered by the acquisition of drug resistance. Therefore, combination therapy that includes BRAFV600E inhibition and avoids resistance is a clinical need. Methods: ATC cell lines 8505C (BRAFV600E/mt), SW1736 (BRAFV600E/mt), KAT18 (BRAFV600E/wt) and Cal-62(BRAFV600E/wt) cells were used in the study. The ability of S100A knockout or /and in combination with the BRAF inhibitor vemurafenib on growth, apoptosis, invasion and apoptosis in ATC cells in vitro was demonstrated by MTT and BrdUrd incorporation assay, Annexin-V-FITC staining analyzed by flow cytometry, Transwell migration and Matrigel invasion assay. S100A4,pERK1/2, pAKT and pROCK1/2 protein was detected by western blot assay; Small molecule inhibitors of Y27632, U0126, MK-2206 and constitutively active forms of pCDNA-Myc-pERK, pCMV6-HA-Akt, pCMV-RhoA were employed, and the mechanistic studies were performed. We assessed the efficiency of in vivo combination treatment with S100A4 knockout and Vemurafenib on tumors. Results: S100A4 knockout induced apoptosis and reduced proliferation by inactivation of pAKT and pERK signals, and inhibited invasion and migration by inactivation of pAKT and RhoA/ROCK1/2 signals in 8505C or Cal-62 cells in vitro, and vice versa in SW1736 and KAT18 cells. Vemurafenib did not affect apoptosis of both 8505C and SW1736 cells, but reduced proliferation via arresting cell cycle, and promoted cell migration and invasion in vitro. Combination treatment with S100A4 knockdown and vemurafenib reduced cell proliferation, migration and invasion in vitro compared to the S100A4 knockdown or Vemurafenib alone. Vemurafenib treatment resulted in a transient inhibition of pERK expression and gradually activation of pAKT expression, but quickly recovery from ERK1/2 activation inhibition by vemurafenib treatment in 4 h for SW1736 and 8505C cells. Combined treatment completely inhibited ERK1/2 and AKT activation during 48 h. In an in vivo mouse model of SW1736 and 8505C, vemurafenib treatment alone did not significantly inhibit tumor growth in both of the tumors, but inhibited tumor growth in combined groups. Conclusion: Our results show S100A4 knockout alone inhibits ATC cells (rich endogenous S100A4) survival and invasion, regardless of the BRAFV600E status, and potentiates the effect of vemurafenib on tumor regression in vitro and in vivo. In addition, S100A4 knockout potently inhibits the recovery from ERK1/2 activation inhibition and the AKT activation following vemurafenib treatment and reversed the vemurafenib resistance. This therapeutic combination may be of benefit in patients with ATC

Association of polymicrobial interactions with dental caries development and prevention

Dental caries is a common oral disease. In many cases, disruption of the ecological balance of the oral cavity can result in the occurrence of dental caries. There are many cariogenic microbiota and factors, and their identification allows us to take corresponding prevention and control measures. With the development of microbiology, the caries-causing bacteria have evolved from the traditional single Streptococcus mutans to the discovery of oral symbiotic bacteria. Thus it is necessary to systematically organized the association of polymicrobial interactions with dental caries development. In terms of ecology, caries occurs due to an ecological imbalance of the microbiota, caused by the growth and reproduction of cariogenic microbiota due to external factors or the disruption of homeostasis by one’s own factors. To reduce the occurrence of dental caries effectively, and considering the latest scientific viewpoints, caries may be viewed from the perspective of ecology, and preventive measures can be taken; hence, this article systematically summarizes the prevention and treatment of dental caries from the aspects of ecological perspectives, in particular the ecological biofilm formation, bacterial quorum sensing, the main cariogenic microbiota, and preventive measures

Genome-wide analysis of WD40 protein family and functional characterization of BvWD40-82 in sugar beet

Sugar beet is one of the most important sugar crops in the world. It contributes greatly to the global sugar production, but salt stress negatively affects the crop yield. WD40 proteins play important roles in plant growth and response to abiotic stresses through their involvement in a variety of biological processes, such as signal transduction, histone modification, ubiquitination, and RNA processing. The WD40 protein family has been well-studied in Arabidopsis thaliana, rice and other plants, but the systematic analysis of the sugar beet WD40 proteins has not been reported. In this study, a total of 177 BvWD40 proteins were identified from the sugar beet genome, and their evolutionary characteristics, protein structure, gene structure, protein interaction network and gene ontology were systematically analyzed to understand their evolution and function. Meanwhile, the expression patterns of BvWD40s under salt stress were characterized, and a BvWD40-82 gene was hypothesized as a salt-tolerant candidate gene. Its function was further characterized using molecular and genetic methods. The result showed that BvWD40-82 enhanced salt stress tolerance in transgenic Arabidopsis seedlings by increasing the contents of osmolytes and antioxidant enzyme activities, maintaining intracellular ion homeostasis and increasing the expression of genes related to SOS and ABA pathways. The result has laid a foundation for further mechanistic study of the BvWD40 genes in sugar beet tolerance to salt stress, and it may inform biotechnological applications in improving crop stress resilience

EGFL6, a potential novel ligand of EGFR, plays roles in cancer metastasis by inducing cancer cell epithelial-mesenchymal transition in nasopharyngeal carcinoma

abstractClinical OncologyDoctoralDoctor of Philosoph

Optical Fiber Sensors and Their Applications

Several novel fiber-based sensors and technologies developed are presented here, including fiber Bragg grating (FBG) based sensors, photonic crystal fiber (PCF) based sensors, specialty fiber-based sensors and distributed fiber sensing systems

Compressive strength of fly ash based geopolymer utilizing waste completely decomposed granite

An attempt to recycle construction waste soils in southern cities of China was presented in this study, where completely decomposed granite (CDG) was utilized to replace fly ash in the production of environment-friendly low-carbon geopolymers. Five crucial parameters were identified and incorporated into the experimental design using the Taguchi method, including fine and gravel particle content of CDG, fly ash content, water-binder ratio, and curing temperature. Test results demonstrate that the newly formulated geopolymer can achieve a compressive strength of up to 14 MPa with a relatively high usage of CDG. The strength initially increases and then stabilizes with an increase in fine particle content. However, no significant relationship is observed between the compressive strength and gravel particle content. Higher curing temperatures and greater fly ash content lead to increased strength, while the strength initially decreases and then slightly increases with changes in the water-binder ratio. To examine the significance of these factors, Analysis of variance (ANOVA) were conducted and a correlation heatmap was drawn, revealing the following order of significance: curing temperature > fly ash content > water-binder ratio > fine particle content > gravel particle content. Furthermore, a robust strength prediction model was modified through multiple regression analysis. Another two additional groups of tests were performed to validate the proposed model, demonstrating the model's strong performance, particularly for curing ages beyond 14 days

Optimization of Fermentation Conditions for the Production of Angiotensin-Converting Enzyme (ACE) Inhibitory Peptides from Cow Milk by Lactobacillus bulgaricus LB6

The purpose of this research was to screen out the optimal -producing peptide conditions for cow milk fermented by Lactobacillus bulgaricus LB6. The effects of temperature, inoculation size, time and skim milk concentration on the ACE inhibition rate of fermented milk were investigated by single factor experiment, and the optimal fermentation conditions were determined by orthogonal experiment. The conditions of the single factor experiment were: Temperatures were 37° C, 39° C, 42° C, 44° C and 46° C. The inoculation amount was 1%, 3%, 5%, 7% and 9%, the time was 8h and 10h. At 12h, 14h and 16h, the concentration of skim milk was 8%, 10%, 12%, 14% and 16%, respectively. The results showed that the optimal fermentation conditions for ACE inhibitory peptide produced by Lactobacillus bulgaricus LB6 were 4% inoculation, 13h in time, 42°C in temperature and 13% in skim milk. Under this condition, the ACE inhibition rate reached 76.50% and the OD value was 0.330. The titration acidity was 116.4°T, the pH was 4.62, and the sensory evaluation was 75 scores