Modulation of Vascular Smooth Muscle Cell Multiplication, Apoptosis, and Inflammatory Damage by miR-21 in Coronary Heart Disease

This study is aimed at exploring the role and potential molecular mechanism of microRNA-21 (miR-21) in coronary heart disease (CHD). RT-qPCR analysis was conducted to detect the expression of miR-21, Sprouty 1 (SPRY1), and connexin 43 (CX43). The protein expression of SPRY1 and CX43 was measured by western blot. ELISA was performed for measuring inflammatory factors, including intercellular adhesion molecule-1 (ICAM-1) and interleukin-1 beta (IL-1β). The target relationship between miR-21 and SPRY1 was determined by dual-luciferase reporter assay. Cell multiplication and apoptosis were detected using CCK-8 assay and flow cytometry analysis, respectively. Our results indicated that miR-21, CX43, and the level of inflammatory cytokines including ICAM-1 and IL-1β were upregulated, while SPRY1 was downregulated in blood samples from CHD patients compared with the controls. Besides, miR-21 directly targeted SRPY-1. miR-21 could suppress SPRY1 expression and enhance CX43 expression in VSMCs. Moreover, miR-21 accelerated cell multiplication and attenuated cell apoptosis in VSMCs. Collectively, these findings suggested that miR-21 could effectively elevate VSMC multiplication and repress apoptosis by targeting SPRY1 in CHD, providing a potential target for therapeutic strategy of CHD.

Biodegradable 3D Printed Scaffolds of Modified Poly (Trimethylene Carbonate) Composite Materials with Poly (L-Lactic Acid) and Hydroxyapatite for Bone Regeneration

Biodegradable scaffolds based on biomedical polymeric materials have attracted wide interest in bone transplantation for clinical treatment for bone defects without a second operation. The composite materials of poly(trimethylene carbonate), poly(L-lactic acid), and hydroxyapatite (PTMC/PLA/HA and PTMC/HA) were prepared by the modification and blending of PTMC with PLA and HA, respectively. The PTMC/PLA/HA and PTMC/HA scaffolds were further prepared by additive manufacturing using the biological 3D printing method using the PTMC/PLA/HA and PTMC/HA composite materials, respectively. These scaffolds were also characterized by Fourier transform infrared spectroscopy (FT-IR), gel permeation chromatography (GPC), automatic contact-angle, scanning electronic micrographs (SEM), diffraction of X-rays (XRD), differential scanning calorimetry (DSC), and thermogravimetry (TG). Subsequently, their properties, such as mechanical, biodegradation, cell cytotoxicity, cell compatibility in vitro, and proliferation/differentiation assay in vivo, were also investigated. Experiment results indicated that PTMC/PLA/HA and PTMC/HA scaffolds possessed low toxicity, good biodegradability, and good biocompatibility and then enhanced the cell multiplication ability of osteoblast cells (MC3T3-E1). Moreover, PTMC/PLA/HA and PTMC/HA scaffolds enhanced the adhesion and proliferation of MC3T3-E1 cells and enabled the bone cell proliferation and induction of bone tissue formation. Therefore, these composite materials can be used as potential biomaterials for bone repatriation and tissue engineering.

miR-206 Inhibits Laryngeal Carcinoma Cell Multiplication, Migration, and Invasion

Laryngeal carcinoma (LC) is one of the common human cancer types. MicroRNAs (miRNAs) were reported to be the essential regulators in cancer diagnosis, treatment, and prognosis. It was reported that miR-206 expression was reduced in various neoplastic diseases. However, the role and functional mechanism of miR-206 in LC progression remain unclear. In this research, miR-206 was found to be associated with tumor-node-metastasis (TNM) staging. In addition, the area under the curve (AUC) of miR-206 was 0.902 for diagnosis of LC and 0.854 for differential diagnosis of stage I-II and stage III-IV patients. Low expression of miR-206 was associated with poor prognosis of LC patients. miR-206 expression was an independent factor affecting the prognosis of LC patients, as revealed by the Cox regression analysis. In vitro experiments demonstrated that miR-206 overexpression reduced cell multiplication, invasion, and migration and increased cell apoptosis in LC cells. Moreover, SOX9 was a target of miR-206, and miR-206 negatively regulated SOX9 expression. Collectively, miR-206 might be a promising biomarker with diagnostic and prognostic value for LC, and the miR-206/SOX9 axis might be a candidate target for LC therapy.

Hsa_circ_0000520 is Involved in Breast Cancer Progression by Targeting miR-542-3p/S1PR1 Axis

Background: CircRNAs feature prominently in breast cancer (BC) progression. This study was intended to investigate the role of hsa_circ_0000520 in BC progression.Methods: After the sample collection, quantitative real-time polymerase chain reaction (qRT-PCR) was conducted for quantifying the expressions of circ_0000520, miR-542-3p, and sphingosine-1-phosphate receptor 1 (S1PR1) mRNA. 5‐Ethynyl‐2′‐Deoxyuridine (EdU) and cell counting kit-8 (CCK-8) assays were used for measuring cell proliferation. Transwell assays were employed to detect cell migration and invasion. Western blotting was utilized for analyzing S1PR1 protein expression. Dual-luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were used to delve into the targeting relationship between circ_0000520 and miR-542-3p.Results: Circ_0000520 expression was markedly elevated in BC cell lines and tissues, and knockdown of circ_0000520 could inhibit BC cell multiplication, migration, and invasion. Circ_0000520 could target miR-542-3p to negatively regulate S1PR1 expression. S1PR1 overexpression plasmid could counteract the inhibitory effects of circ_0000520 knockdown on BC cell proliferation, migration, and invasion.Conclusion: Circ_0000520, as a cancer-promoting circRNA, participates in BC progression by regulating miR-542-3p/S1PR1 axis.

NIMG-29. ELEVATION OF GLUTAMINE AND CITRATE BY MR SPECTROSCOPY IS AN IMAGING BIOMARKER OF RAPID CELL PROLIFERATION IN GLIOMAS

Prior studies suggested glutamine metabolism in cancers may be altered to meet the high demands of nucleotide biosynthesis in cancers. We conducted 1H MR spectroscopy in 18 glioma patients and analyzed the metabolic data together with post-gadolinium MRI and cell proliferation rate (MIB-1 label index). The optimized MRS (TE 97 ms PRESS) provided well discernible signal of glutamine at 2.45 ppm and a negative-polarity signal of citrate at 2.6 ppm, without considerable overlaps with neighboring signals. The 18 patients had biopsy-proven anaplastic gliomas or glioblastomas. Concurrent elevation of glutamine and citrate was identified in 15 gliomas. These gliomas presented with enhancement in post-gadolinium MRI, indicative of broken blood-brain barrier (BBB). The 15 gliomas were grouped as subset-1 while the other 3 gliomas that had elevated citrate without elevation of glutamine were grouped as subset-2. Citrate level was significantly different between the two subsets of gliomas (1.4+/-0.5 vs. 1.6+/-0.4 mM). However, subst-1 had significantly higher choline (4.7+/-1.8 vs. 1.6+/-0.3 mM, p< 0.01) and higher MIB-1 compared with subset-2. Given that choline is a cellularity marker, a finding of high choline and high MIB-1 with elevation of glutamine and citrate suggests that the tumors have high tumor cellularity and cell multiplication competence. Of the 18 gliomas, 13 were IDH mutated with elevated 2HG. The glutamine and citrate levels in IDH-mutant gliomas were not significantly different from those in IDH wildtype gliomas. In conclusion, high-grade gliomas undergo a metabolic rearrangement of concurrent elevation of glutamine and citrate to attain malignant characters such as high cellularity, rapid cell proliferation, and BBB breakdown. IDH mutant and IDH wildtype gliomas may share a common metabolic rearrangement of glutamine-mediated citrate elevation to attain malignancy. Measurements of glutamine and citrate may serve a potential imaging biomarker for aggressive gliomas.

UBTF facilitates melanoma progression via modulating MEK1/2-ERK1/2 signalling pathways by promoting GIT1 transcription

Background UBTF is an HMGB-box DNA binding protein and a necessary Pol I/Pol II basal transcription factor. It has been found that UBTF involves in carcinogenesis and progression of a few cancers. Nevertheless, the the biological function and potential molecular mechanism of UBTF in melanoma are still not clear and need to be clarified. Methods UBTF and GIT1 expressions in melanoma specimens and cell lines were examined by quantitative real-time PCR (qRT-PCR) and Western blot. MTT and colony formation assays were used to investigate the effects of UBTF and GIT1 on melanoma cell proliferation. Cell cycle and apoptosis assays were detected by flow cytometry. Tumor formation assay was used to analyze the effect of UBTF on melanoma growth. Bioinformatics predicting, chromatin immunoprecipitation (ChIP)-qRT-PCR and reporter gene assay were fulfilled for verifing GIT1 as UBTF targeting gene. Results Here we reported that UBTF mRNA and protein expressions were upregulated in primary melanoma specimens and cell lines. UBTF overexpression facilitated melanoma cell proliferation and cell cycle progression and restrained. Silencing UBTF suppressed cell multiplication, cell cycle progression and tumor growth, and promoted apoptosis. UBTF expression was positively related with GIT1 expression in human melanoma tissues. It was verified that UBTF promoted GIT1 transcription in melanoma cells through binding to the promoter region of GIT1. Furthermore, GIT1 overexpression promoted melanoma cell growth and suppressed apoptosis. Knockdown of GIT1 inhibited cell multiplication and induced apoptosis. Overexpression of GIT1 eliminated the effects of silencing UBTF on melanoma cells. Importantly, UBTF activated MEK1/2-ERK1/2 signalling pathways by upregulating GIT1 expression. Conclusions Our study demonstrates that UBTF promotes melanoma cell proliferation and cell cycle progression by promoting GIT1 transcription, thereby activating MEK1/2-ERK1/2 signalling pathways. The findings indicate that UBTF plays a crucial function in melanoma and may be a potential therapeutic target for the treatment of this disease.

Novel Computer Aided Diagnostic System Using Synergic Deep Learning Technique for Early Detection of Pancreatic Cancer

Pancreatic cancer (PC) in the more extensive sense alludes to in excess of 277 distinct kinds of cancer sickness. Researchers have recognized distinctive phase of pancreatic cancers, showing that few quality transformations are engaged with cancer pathogenesis. These quality transformations lead to unusual cell multiplication. Therefore, in this study we propose a Computer Aided Diagnosis (CAD) system using Synergic Inception ResNet-V2, Deep convoluted neural network architecture for the identification of PC cases from publically Usable CT images that could extract PC graphical functionality to include clinical diagnosis before the pathogenic examination, saving valuable time for disease prevention. Simulation results using MATLAB is shown to illustrate that quite promising results have been obtained in terms of accuracy in detecting patients infected with BC. Accuracy of 99.23 per cent is reached using the proposed deep learning method, which is better than all other state-of-the-art approaches available in the literature. The calculation time was found to be less than the other current 22 second process. The proximity of the suggested approach to the True Positive values in the ROC curve suggests a result that is greater than the other methods. The comparative study with Inception ResNet-V2 is based on separate test and training data at a rate of 90 percent-10 percent, 80 percent-20 percent and 70 percent-30% respectively, which shows the robustness of the proposed research work. Experimental findings show the proposed reliability of the device relative to other detection approaches. The proposed CAD device is fully automated and has thus proved to be a promising additional diagnostic tool for frontline clinical physicians.

​Regeneration of Stem Cuttings of Pomegranate cv. Bhagwa as Influenced by PGR’s and Planting Time

Background: Pomegranate is being cultivated in the tropical and sub-tropical parts of the world for its delicious fruits. It can be propagated from seeds as well as from softwood, semi-hardwood and hardwood cuttings. The application of auxins encourages rooting in stem cutting owing to their ability to active cambium regeneration, cell division and cell multiplication. To meet the demand of pomegranate in the current situation and owing to its importance, the present study was undertaken. Methods: For regeneration of stem cuttings of pomegranate cv. Bhagwa as influenced by PGR‘s and planting time a study was conducted at the Department of Horticulture, Khalsa College, Amritsar during 2019-2020. The trial was undertaken with ten treatments comprising of IBA, PHB and NAA (500, 750 and 1000 ppm) each by quick dip method along with control planted in the first week of August and January. Result: The results of the study revealed that IBA 1000 ppm (T3) proved to be the best in terms of minimum days to first sprouting (11.06), maximum sprouting (91.10%), rooting (83.31%), number of roots per cutting (60.03), root length (11.55 cm), fresh weight of roots (1.08 g) and dry weight of roots (0.55 g). Among planting time the cuttings planted on first week of January proved to be effective in terms of sprouting (87.83%), survival (84.70%), rooting (78.81%) and number of roots (52.06) respectively.

Temporal Evolution of the Gravitaxis of Euglena gracilis from a Single Cell

Gravitaxis is one of the most important issues in the growth of microalgae in the water column; it determines how easily cells receive sunlight with a comfortable intensity that is below the damaging threshold. We quantitatively investigated and analyzed the gravitaxis and cell multiplication of Euglena gracilis using vertically placed microchambers containing a single cell. A temporal change in gravitaxis and cell multiplication was observed after transferring the cells to fresh culture medium for 9 days. We performed 29 individual experiments with 2.5 × 2.5 × 0.1 mm square microchambers and found that the cells showed positive, negative, and moderate gravitaxis in 8, 7, and 14 cases, respectively, after transferring to fresh culture medium. A common trend was observed for the temporal change in gravitaxis for the eight initially positive gravitaxis cases. The cells with initially positive gravitaxis showed a higher rate of cell multiplication than those with initially negative gravitaxis. We also discussed the gravitaxis mechanism of E. gracilis from the observed trend of gravitaxis change and swimming traces. In addition, bioconvection in a larger and thicker chamber was investigated at a millimeter scale and visualized.