CircROR1 upregulates CCNE1 expression to promote melanoma invasion and metastasis by recruiting KAT2A.

Circular RNAs (circRNAs) play important roles in cancer occurrence and progression. To explore and elucidate the clinical significance of specific circular RNA in melanoma and its potential molecular mechanism. CircROR1 expression in melanoma cells and tissues was confirmed by qRT-PCR and ISH. qRT-PCR and Western blotting were performed to measure the levels of CCNE1, KAT2A, MMP9 and TIMP2. MTT, Transwell and wound healing assays were performed to evaluate cell proliferation, invasion and metastasis. A xenograft mouse model was established to further verify the CircROR1/CCNE1 axis in vivo. RNA pull-down and RIP assays were performed to detect the direct interaction KAT2A and CircROR1. A ChIP assay was used to investigate the enrichment of H3K9ac acetylation in the CCNE1 promoter. CircROR1 was significantly upregulated in metastatic melanoma cells and tissues, promoting proliferation, invasion and metastasis in vitro and tumour growth in vivo. CircROR1 overexpression increased CCNE1 and MMP9 protein expression and decreased TIMP2 protein expression. Functional rescue assays demonstrated that CircROR1 played a role in promoting malignant progression through CCNE1. CircROR1 specifically bound to the KAT2A protein without affecting its expression. CircROR1 overexpression increased the level of H3K9ac modification in the CCNE1 promoter region by recruiting KAT2A, thus upregulating CCNE1 expression. CircROR1 upregulates CCNE1 expression through KAT2A-mediated histone acetylation. Our research confirms the critical role of CircROR1 in melanoma invasion and metastasis, and CircROR1 could serve as a potential therapeutic target for melanoma treatment.

Study of the prediction of gamma passing rate in dosimetric verification of intensity-modulated radiotherapy using machine learning models based on plan complexity.

Objective: To predict the gamma passing rate (GPR) in dosimetric verification of intensity-modulated radiotherapy (IMRT) using three machine learning models based on plan complexity and find the best prediction model by comparing and evaluating the prediction ability of the regression and classification models of three classical algorithms: artificial neural network (ANN), support vector machine (SVM) and random forest (RF). Materials and methods: 269 clinical IMRT plans were chosen retrospectively and the GPRs of a total of 2340 fields by the 2%/2mm standard at the threshold of 10% were collected for dosimetric verification using electronic portal imaging device (EPID). Subsequently, the plan complexity feature values of each field were extracted and calculated, and a total of 6 machine learning models (classification and regression models for three algorithms) were trained to learn the relation between 21 plan complexity features and GPRs. Each model was optimized by tuning the hyperparameters and ten-fold cross validation. Finally, the GPRs predicted by the model were compared with measured values to verify the accuracy of the model, and the evaluation indicators were applied to evaluate each model to find the algorithm with the best prediction performance. Results: The RF algorithm had the optimal prediction effect on GPR, and its mean absolute error (MAE) on the test set was 1.81%, root mean squared error (RMSE) was 2.14%, and correlation coefficient (CC) was 0.72; SVM was the second and ANN was the worst. Among the classification models, the RF algorithm also had the optimal prediction performance with the highest area under the curve (AUC) value of 0.80, specificity and sensitivity of 0.80 and 0.68 respectively, followed by SVM and the worst ANN. Conclusion: All the three classic algorithms, ANN, SVM, and RF, could realize the prediction and classification of GPR. The RF model based on plan complexity had the optimal prediction performance which could save valuable time for quality control workers to improve quality control efficiency.

Dosimetric verification of stereotactic body radiotherapy treatment plan via ArcCHECK-3DVH system. / ArcCHECK-3DVH系统在体部立体定向放射治疗剂量验证中的应用.

OBJECTIVES: To study the feasibility of ArcCHECK-3DVH system in dosimetric verification for stereotactic body radiaotherapy (SBRT) with flattening filter free (FFF) model. METHODS: SBRT treatment plans for 57 patients were introduced into ArcCHECK phantom and recalculated. The calculated dose distribution of treatment planning system and the measured dose distribution of ArcCHECK phantom were compared by γ analysis. Then the 3 dimensional dose distribution of target and organs at risk was reconstructed by 3DVH software. The reconstructed dose and calculated dose with treatment planning system (TPS) were compared, and the dose volume γ pass rate and deviation of dose volume parameters to the target and organs at risk were quantitatively valuated. RESULTS: Based on the threshold criteria (3%, 3 mm, 10%), namely the deviation of measuring points between the planned value and the measured value was less than 3%, and the proportion of points with similar values in the plane or sphere with the center of the point and the radius of 3 mm was 10%, the relative and absolute dose pass rates of SBRT treatment plans in ArcCHECK system via γ analysis were greater than 95%. Based on the stricter threshold criteria (2%, 2 mm, 10%), the relative and absolute dose pass rates of SBRT treatment plan in ArcCHECK system via γ analysis were about 93%. In 3DVH dose verification, the γ pass rate of target and organs at risk was exceed 97%, and the deviations in 3DVH of the target and organs at risk were less than ±5%. CONCLUSIONS: The ArcCHECK-3DVH system in dose verification can provide more comprehensive dose distribution information to reasonably evaluate the SBRT plan, with more significance for guiding clinical treatment.

Micro Magnetic Field Produced by Fe3O4 Nanoparticles in Bone Scaffold for Enhancing Cellular Activity.

The low cellular activity of poly-l-lactic acid (PLLA) limits its application in bone scaffold, although PLLA has advantages in terms of good biocompatibility and easy processing. In this study, superparamagnetic Fe3O4 nanoparticles were incorporated into the PLLA bone scaffold prepared by selective laser sintering (SLS) for continuously and steadily enhancing cellular activity. In the scaffold, each Fe3O4 nanoparticle was a single magnetic domain without a domain wall, providing a micro-magnetic source to generate a tiny magnetic field, thereby continuously and steadily generating magnetic stimulation to cells. The results showed that the magnetic scaffold exhibited superparamagnetism and its saturation magnetization reached a maximum value of 6.1 emu/g. It promoted the attachment, diffusion, and interaction of MG63 cells, and increased the activity of alkaline phosphatase, thus promoting the cell proliferation and differentiation. Meanwhile, the scaffold with 7% Fe3O4 presented increased compressive strength, modulus, and Vickers hardness by 63.4%, 78.9%, and 19.1% compared with the PLLA scaffold, respectively, due to the addition of Fe3O4 nanoparticles, which act as a nanoscale reinforcement in the polymer matrix. All these positive results suggested that the PLLA/Fe3O4 scaffold with good magnetic properties is of great potential for bone tissue engineering applications.

In Situ Generation of Hydroxyapatite on Biopolymer Particles for Fabrication of Bone Scaffolds Owning Bioactivity.

Hydroxyapatite (HAP) can endow a biopolymer scaffold with good bioactivity and osteoconductive ability, while the interfacial bonding is fairly weak between HAP and biopolymers. In this study, HAP was in situ generated on poly(l-lactic acid) (PLLA) particles, and then they were used to fabricate a scaffold by selective laser sintering. Detailedly, PLLA particles were first functionalized by dopamine oxide polymerization, which introduced abundance active catechol groups on the particle surface, and subsequently, the catechol groups concentrated Ca2+ ions by chelation in a simulated body fluid solution, and then, Ca2+ ions absorbed PO43- ions through electrostatic interactions for in situ nucleation of HAP. The results indicated that HAP was homogeneously generated on the PLLA particle surface, and HAP and PLLA exhibited good interfacial bonding in the HAP/PLLA scaffolds. Meanwhile, the scaffolds displayed excellent bioactivity by inducing apatite precipitation and provided a good environment for human bone mesenchymal stem cell attachment, proliferation, and osteogenic differentiation. More importantly, the ingrowth of blood vessel and the formation of new bone could be stimulated by the scaffolds in vivo, and the bone volume fraction and bone mineral density increased by 44.44 and 41.73% compared with the pure PLLA scaffolds, respectively. Serum biochemical indexes fell within the normal range, which indicated that there was no harmful effect on the normal functioning of the body after implanting the scaffold.

Bioactive Tetracalcium Phosphate Scaffolds Fabricated by Selective Laser Sintering for Bone Regeneration Applications.

Tetracalcium phosphate (TTCP), a potential biological scaffold material, has attracted increasing interest for bone regeneration applications due to its good biodegradability and biocompatibility. In this research, three-dimensional porous TTCP scaffolds were manufactured via selective laser sintering (SLS), and an in-depth and meticulous study on the influence of laser power on the microstructure and mechanical properties of TTCP scaffolds was performed. The results showed that the TTCP particles fused together and formed a solid object due to the decrease in the number of micro-pores in the scaffold as the laser power increased from 6 W to 9 W. The maximum compressive strength that the scaffold could withstand and the strength of the fracture toughness were 11.87 ± 0.64 MPa and 1.12 ± 0.1 MPa·m1/2, respectively. When the laser power increased from 9 W to 10 W, the TTCP grains grew abnormally, resulting in diminished mechanical properties. The bioactivity tests showed that the surfaces of the scaffolds were entirely covered by bone-like apatite layers after soaking in simulated body fluid (SBF) for three days, indicating that the scaffolds exhibit excellent bioactivity. Moreover, cell experiments showed that the TTCP scaffolds had good biocompatibility. This study indicated that SLS-fabricated TTCP scaffolds may be a promising candidate for bone regeneration applications.

TiO2-Induced In Situ Reaction in Graphene Oxide-Reinforced AZ61 Biocomposites to Enhance the Interfacial Bonding.

Graphene oxide (GO) can improve the degradation resistance of biomedical Mg alloy because of its excellent impermeability and outstanding chemical inertness. However, the weak interfacial bonding between GO and Mg matrix leads to easily detaching during degradation. In this study, in situ reaction induced by TiO2 took place in the AZ61-GO biocomposite to enhance the interfacial bonding between GO and Mg matrix. For the specific process, TiO2 was uniformly and tightly deposited onto the GO surface by hydrothermal reaction (TiO2/GO) first and then used for fabricating AZ61-TiO2/GO biocomposites by selective laser melting (SLM). Results showed that TiO2 was in situ reduced by magnesiothermic reaction during SLM process, and the reduzate Ti, on the one hand, reacted with Al in the AZ61 matrix to form TiAl2 and, on the other hand, reacted with GO to form TiC at the AZ61-GO interface. Owing to the enhanced interfacial bonding, the AZ61-TiO2/GO biocomposite showed 12.5% decrease in degradation rate and 10.1% increase in compressive strength as compared with the AZ61-GO biocomposite. Moreover, the AZ61-TiO2/GO biocomposite also showed good cytocompatibility because of the slowed degradation. These findings may provide guidance for the interfacial enhancement in GO/metal composites for biomedical applications.

[Dosimetric verification of flattening filter free model based on TrueBeam accelerator using ArcCheck system].

OBJECTIVE: To study the feasibility of ArcCheck verification system in dosimetric verification for stereotactic radiotherapy (SRT) the stereotactic radiotherapy (SRT) with flattening filter free (FFF) model.
 Methods: A total of 76 cases under SRT treatment plans were introduced into ArcCheck phantom and recalculated. Threshold criteria was set as (3%, 3 mm, 10%) or (2%, 2 mm, 10%). The calculated dose distribution and the measured dose distribution of ArcCheck phantom were compared by means of distance to agree (DTA) and Gamma analysis method respectively.
 Results: Based on the threshold criteria (3%, 3 mm, 10%), the relative and absolute mean pass rates of SRT treatment plans by DTA and Gamma analysis were greater than 95%. Based on the threshold criteria (2%, 2 mm, 10%), the relative and absolute mean pass rates of SRT treatment plan by DTA and Gamma analysis were about 90%. The dose pass rate of Gamma analysis method was slightly higher than that of DTA analysis method (P<0.001).
 Conclusion: The ArcCheck verification system is a rapid and accurate method for SRT dose verification, and discrepancies are found in different analysis methods.

[Dosimetric comparison of intensity-modulated arc radiotherapy and fixed beam dynamic intensity-modulated radiation therapy in nasopharyngeal carcinoma].

OBJECTIVE: To compare the dosimetric differences of dosiology between intensity-modulated arc radiotherapy (IMAT) and dynamic intensity-modulated radiation therapy (dIMRT) in nasopharyngeal carcinoma. METHODS: CT data from 25 patients treated in our radiotherapy center were selected randomly for this study. For each patient, the IMAT technique and the fixed beam dIMRT technique were accomplished by the simultaneously integrated boost. Dose volume histogram (DVH) data, isodose distribution, monitor units (MUs) and treatment time were compared in the two techniques. RESULTS: There was no significant difference between the IMAT and the dIMRT in dose received by 95% of target volumes (D(95)) (P>0.05). Overall, the mean dose (D(mean)), maximal dose (D(max)) and volume percentage receiving at least of 107% of the prescribed dose (V(107%)) of planning target volume (PTV) for the IMAT were increased slightly ,compared with the dlMRT (P<0.05). There were no significant differences in dosimetric indices of organs at risk (OARs) including spinal cord,optical nerves,lens and temporomandibular joints in the two techniques (P>0.05). Compared with the dlMRI, the D(max) of brain stem for the IMAT was increased slightly (P<0.05). Similar trends was observed for the D(mean) and dose received by 50% of volume (D(50)) of the left and right parotid glands (P<0.05). Healthy tissue (defined as the volume of the body minus PTV,B-P) irradiated from 800 cGy in the IMAT was higher, and that from 1200-4500 cGy was lower compared with the dlMRI (P<0.05).The average number of MUs was reduced by 62.7% per fraction, and the treatment time was on average reduced by 60.1% per fraction in the IMAT compared with the dlMRI. CONCLUSION: There is a slight difference in dosiology between the two radiotherapy techniques investigated, but they both meet the clinical requirement. Compared with the dIMRT, the IMAT delivers less irradiation to healthy tissue, uses fewer MUs and takes less time during radiotherapy for nasopharyngeal carcinoma.