Validation of the "obturator functioning scale" for Chinese-speaking patients with obturator prostheses after cancer-related maxillectomy.

Objective: The Obturator Functioning Scale (OFS) is a scale without formal measures of validity in any language. This study aimed to translate and adapt the OFS from English to Chinese and check its reliability and validity in Chinese-speaking patients with obturator prostheses after cancer-related maxillectomy. Methods: The 15-item Chinese preversion of the OFS was completed by 133 patients in three tertiary stomatological hospitals. Of these, 41 completed it again one week after the first measurement. The patients also completed the Chinese version of the University of Washington quality of life scale (UW-QOL, Version 4). Results: Item 12 ("upper lip feels numb") was deleted to achieve a better statistical fit. The 14-item Chinese version of the OFS (OFS-Ch) demonstrated high internal consistency (Cronbach's alpha = 0.908). The test-retest reliability coefficients for most items exceeded 0.90, indicating substantial reproducibility. Confirmatory factor analysis found that the scale consisted of three correlated factors: 1) eating (four items), 2) speech (five items), and 3) other problems (five items). This explained 70.2 % of the total variance using exploratory factor analysis. The scale was significantly convergent and discriminant and could validly discriminate between patients with Brown I and IId maxillary defects. Conclusions: Our results showed that the OFS-Ch scale is a valid tool for evaluating oral dysfunction and satisfaction with appearance for patients with the obturator prosthesis and identifying those at risk of poor obturator function in clinical settings.

Kaempferol attenuates inflammation in lipopolysaccharide-induced gallbladder epithelial cells by inhibiting the MAPK/NF-κB signaling pathway.

Kaempferol (KPR), a flavonoid compound found in various plants and foods, has garnered attention for its anti-inflammatory, antioxidant, and anticancer properties. In preliminary studies, KPR can modulate several signaling pathways involved in inflammation, making it a candidate for treating cholecystitis. This study aimed to explore the effects and mechanisms of KPR on lipopolysaccharide (LPS)-induced human gallbladder epithelial cells (HGBECs). To assess the impact of KPR on HGBECs, the HGBECs were divided into control, KPR, LPS, LPS + KPR, and LPS + UDCA groups. Cell viability and cytotoxicity were evaluated by MTT assay and lactate dehydrogenase (LDH) assay, respectively, and concentrations of KPR (10-200 µM) were tested. LPS-induced inflammatory responses in HGBECs were to create an in vitro model of cholecystitis. The key inflammatory markers (IL-1ß, IL-6, and TNF-α) levels were quantified using ELISA, The modulation of the MAPK/NF-κB signaling pathway was measured by western blot using specific antibodies against pathway components (p-IκBα, IκBα, p-p65, p65, p-JNK, JNK, p-ERK, ERK, p-p38, and p38). The cell viability and LDH levels in HGBECs were not significantly affected by 50 µM KPR, thus it was selected as the optimal KPR intervention concentration. KPR increased the viability of LPS-induced HGBECs. Additionally, KPR inhibited the inflammatory factors level (IL-1ß, IL-6, and TNF-α) and protein expression (iNOS and COX-2) in LPS-induced HGBECs. Furthermore, KPR reversed LPS-induced elevation of p-IκBα/IκBα, p-p65/p65, p-JNK/JNK, p-ERK/ERK, and p-p38/p38 ratios. KPR attenuates the LPS-induced inflammatory response in HGBECs, possibly by inhibiting MAPK/NF-κB signaling.

Magnetic-acoustic actuated spinous microrobot for enhanced degradation of organic pollutants.

A growing interest in the development of efficient strategies for the removal of organic pollutants from polluted water is emerging. As such, artificial micro/nano machines performing excellent water purification tasks have recently attracted more research attention of scientists. Hereby a spinous Fe3O4@PPy microrobot is presented that towards an efficient organic pollutant removal by enhancing Fenton-like reaction. The microrobot is fabricated by wrapping polypyrrole (PPy) on a spiny magnetic template prepared from sunflowers pollen. Modulating the sound pressure and frequency of the ultrasonic field enables the Fe3O4@PPy microrobot to present multimode motion, such as violent eruption-like motion caused by local cavitation (ELM), march-like unific motion (MLM), and typhoon-like rotation toward the center gathered motion (TLM). This multimode motion achieves the sufficient locomotion of microrobots in three-dimensional space and effective contact with organic pollutants in polluted water. Furthermore, a 5.2-fold increase in the degradation rate of methylene blue has been realized using Fe3O4@PPy microrobots under low-concentration hydrogen peroxide conditions. Also, the magnetically controlled recovery of microrobots from water after the completion of the degradation task has been demonstrated. The magnetic-acoustic actuated spinous microrobot can be extrapolated to other catalytic microrobot, developing a new strategy for an easier implementation and recovery of microrobot in real applications of water purification.

Effect of Secondary-Phase Precipitation on Mechanical Properties and Corrosion Resistance of 00Cr27Ni7Mo5N Hyper-Duplex Stainless Steel during Solution Treatment.

In this work, the effect of secondary-phase precipitation on the microstructure, mechanical properties, and corrosion resistance of 00Cr27Ni7Mo5N hyper-duplex stainless steel (HDSS) during solution treatment was investigated. The results reveal that σ-phase precipitates at the interface between the α and γ phase when the solution treatment temperature is lower than 1070 °C. It is not only brittle, but also prone to create a Cr-depleted zone, which significantly deteriorates the mechanical properties and corrosion resistance. With the increase in the solution treatment temperature, the volume fraction of ferrite gradually increases. The yield strength and tensile strength increase slightly, but the elongation decreases. At the same time, the impact toughness shows a trend of first increasing and then decreasing. When the solution treatment temperature is higher than 1130 °C, Cr2N precipitates in the ferrite. The precipitation of Cr2N causes a decrease in the plastic toughness, but it does not deteriorate the mechanical properties as significantly as the σ phase. However, it can also cause the formation of a Cr-depleted zone that significantly decreases the corrosion resistance. There is no secondary-phase precipitation in the sample after solution treatment at 1100 °C, which shows the best mechanical properties and corrosion resistance.

Epitope-based minigene vaccine targeting fibroblast activation protein α induces specific immune responses and anti-tumor effects in 4 T1 murine breast cancer model.

Fibroblast activation protein (FAPα) is a tumor stromal antigen expressed by cancer-associated fibroblasts (CAFs) in more than 90 % of malignant epithelial carcinomas. FAPα-based immunotherapy has been reported and showed that FAPα-specific immune response can remold immune microenvironment and contribute to tumor regression. Many FAPα-based vaccines have been investigated in preclinical trials, which can elicit strong and durable cytolytic T lymphocytes (CTL) with good safety. However, epitope-based FAPα vaccines are rarely reported. To break tolerance against self-antigens, analogue epitopes with modified peptides at the anchor residues are typically used to improve epitope immunogenicity. To investigate the feasibility of a FAPα epitope-based vaccine for cancer immunotherapy in vivo, we conducted a preclinical study to identify a homologous CTL epitope of human and mouse FAPα and obtained its analogue epitope in BALB/c mice, and explored the anti-tumor activity of their minigene vaccines in 4 T1 tumor-bearing mice. By using in silico epitope prediction tools and immunogenicity assays, immunodominant epitope FAP.291 (YYFSWLTWV) and its analogue epitope FAP.291I9 (YYFSWLTWI) were identified. The FAP.291-based epitope minigene vaccine successfully stimulated CTLs targeting CAFs and exhibited anti-tumor activity in a 4 T1 murine breast cancer model. Furthermore, although the analogue epitope FAP.291I9 enhanced FAP.291-specific immune responses, improvement of anti-tumor immunity effects was not observed. Check of immunosuppressive factors revealed that the high levels of IL-10, IL-13, myeloid-derived suppressor cells and iNOS induced by FAP.291I9 increased, which considered the main cause of the failure of the analogue epitope-based vaccine. Thus, we demonstrated for the first time that the FAP.291 minigene vaccine could induce mouse CTLs and also function as a tumor regression antigen, providing the basis for future studies of FAPα epitope-based vaccines. This study may also be valuable for further improvement of the immunogenicity of analogue epitope vaccines.

Magnetic-Driven Hydrogel Microrobots Selectively Enhance Synthetic Lethality in MTAP-Deleted Osteosarcoma.

Background: Drugs based on synthetic lethality have advantages such as inhibiting tumor growth and affecting normal tissue in vivo. However, specific targets for osteosarcoma have not been acknowledged yet. In this study, a non-targeted but controllable drug delivery system has been applied to selectively enhance synthetic lethality in osteosarcoma in vitro, using the magnetic-driven hydrogel microrobots. Methods: In this study, EPZ015666, a PRMT5 inhibitor, was selected as the synthetic lethality drug. Then, the drug was carried by hydrogel microrobots containing Fe3O4. Morphological characteristics of the microrobots were detected using electron microscopy. In vitro drug effect was detected by the CCK-8 assay kit, Western blotting, etc. Swimming of microrobots was observed by a timing microscope. Selective inhibition was verified by cultured tumors in an increasing magnetic field. Results: Genomic mutation of MTAP deletion occurred commonly in pan-cancer in the TCGA database (nearly 10.00%) and in osteosarcoma in the TARGET database (23.86%). HOS and its derivatives, 143B and HOS/MNNG, were detected by MTAP deletion according to the CCLE database and RT-PCR. EPZ015666, the PRMT5 inhibitor, could reduce the SDMA modification and inhibition of tumor growth of 143B and HOS/MNNG. The hydrogel microrobot drug delivery system was synthesized, and the drug was stained by rhodamine. The microrobots were powered actively by a magnetic field. A simulation of the selected inhibition of microrobots was performed and lower cell viability of tumor cells was detected by adding a high dose of microrobots. Conclusion: Our magnetic-driven drug delivery system could carry synthetic lethality drugs. Meanwhile, the selective inhibition of this system could be easily controlled by programming the strength of the magnetic field.

Glutamine deficiency promotes recurrence and metastasis in colorectal cancer through enhancing epithelial-mesenchymal transition.

BACKGROUND: Glutamine is the most abundant amino acid in the body and plays a vital role in colorectal cancer (CRC) cell metabolism. However, limited studies have investigated the clinical and prognostic significance of preoperative serum glutamine levels in patients with colorectal cancer, and the underlying mechanism has not been explored. METHODS: A total of 121 newly diagnosed CRC patients between 2012 and 2016 were enrolled in this study. Serum glutamine levels were detected, and their associations with clinicopathological characteristics, systemic inflammation markers, carcinoembryonic antigen (CEA) and prognosis were analysed. In addition, the effect of glutamine depletion on recurrence and metastasis was examined in SW480 and DLD1 human CRC cell lines, and epithelial-mesenchymal transition (EMT)-related markers were detected to reveal the possible mechanism. RESULTS: A decreased preoperative serum level of glutamine was associated with a higher T-class and lymph node metastasis (P < 0.05). A higher serum level of glutamine correlated with a lower CEA level (r = - 0.25, P = 0.02). Low glutamine levels were correlated with shorter overall survival (OS) and disease-free survival (DFS). Multivariate Cox regression analysis showed that serum glutamine was an independent prognostic factor for DFS (P = 0.018), and a nomogram predicting the probability of 1-, 3- and 5-year DFS after radical surgery was built. In addition, glutamine deficiency promoted the migration and invasion of CRC cells. E-cadherin, a vital marker of EMT, was decreased, and EMT transcription factors, including zeb1and zeb2, were upregulated in this process. CONCLUSIONS: This study elucidated that preoperative serum glutamine is an independent prognostic biomarker to predict CRC progression and suggested that glutamine deprivation might promote migration and invasion in CRC cells by inducing the EMT process.

Role of Circular RNAs in the Pathogenesis of Malignant Melanoma.

Malignant melanoma is one of the most aggressive types of skin cancer. Thus, efficient diagnosis and treatment methods are crucial for advanced melanoma. Circular RNAs (circRNAs) have been regarded as a 'splicing noise' in the past decades. However, several circRNAs have been recently reported to be differentially expressed in melanoma, and the cell or tissue-specific expression makes these suitable candidate diagnostic or therapeutic biomarkers. In addition, emerging studies have confirmed that circular RNAs play pivotal roles in the proliferation, invasion, metastasis, and migration of malignant melanoma. However, specific pathogenic mechanisms between melanoma and circRNAs remain unclear. In the present study, it was summarized that circRNAs are associated with the pathogenesis of melanoma, including hsa_circ_0083444, hsa_circ_0005320, hsa_circ_0067531, hsa_circ_0084043, hsa_circ_0000082, hsa_circ_0016418, hsa_circ_0085533 and hsa_circ_0025039, hsa_circ_0001946, hsa_circ_0002770, hsa_circ_0079593, hsa_circ_0027247, hsa_circ_0017247, hsa_circ_0020710. These can provide potential diagnosis, treatment, and prognostication biomarkers for advanced melanoma in clinical applications.

Radioprotective effects and mechanism of HL-003 on radiation-induced salivary gland damage in mice.

Ionizing radiation (IR) can cause damage to the structure and function of salivary glands. Our research group independently synthesized the ROS scavenger, HL-003. The aim of this study was to explore the protective effects and underlying mechanisms of HL-003 on radiation-induced salivary gland injury. Salivary flow rate measurement, H&E staining, immunohistochemistry, FRAP, TUNEL, and western blotting were used to evaluate the radioprotective effect on salivary glands. The results showed that HL-003 protected the salivary secretion function by protecting the AQP-5 protein, on the salivary epithelial cell membrane, from IR damage. HL-003 reduced oxidative stress in the salivary gland by regulating the expression of ROS-related proteins NOX4, SOD2, and 8-OHdG. Furthermore, HL-003 downregulated the expression of p-p53, Bax, caspase 3, and caspase 9, and upregulated the expression of Bcl-2, suggesting that it could inhibit the activation of p53 to reduce cell apoptosis. In conclusion, HL-003 is an effective radioprotector that prevents damage of the radiation-induced salivary gland.

Preclinical Safety and Biodistribution in Mice Following Single-Dose Intramuscular Inoculation of Tumor DNA Vaccine by Electroporation.

The safety, biodistribution, and pharmacokinetics of any new therapeutic tumor DNA vaccine must be evaluated in preclinical studies. We previously developed the DNA vaccine (CpDV-IL2-sPD1/MUC1 and survivin), which showed excellent antitumor effects in a variety of tumor models. In this study, we demonstrate the safety and biodistribution after immunization with naked DNA vaccine (10 mg/kg) by electroporation in a mice model. All mice reached the end of the study with good body conditions. By established and validated QPCR method, we found high-copy plasmid DNA at the injection site (muscle) on day 1 in all eight animals, followed by a downward trend. By day 49, a small amount of plasmid DNA was still detectable, but only in one mouse. On reproductive safety, no plasmids existed in the ovary at any time point. Also, only two of the 16 testis samples could detect a very small amount of DNA on days 7 and 14. The most important thing was that plasmids were cleared from almost all organs (heart, liver, spleen, lung, kidney, stomach, blood, thymus, intestine) on day 49. In summary, the results of our experiments demonstrate that the DNA vaccine delivered by electroporation was shown to be safe and merits further development for cancer treatment.

B16 Membrane-Coated Vesicles for Combined Photodynamic Therapy and Immunotherapy Shift Immune Microenvironment of Melanoma.

Introduction: Coating of nanomedicine with cell membranes has attracted increasing attention as it can boost biocompatibility and improve the efficiency of treatment. Herein, we prepared innovative tumor cell-membrane-coated vesicles based on photodynamic therapy (PDT) drug indocyanine green (ICG) and explore the effect on melanoma in vitro and in vivo. Methods: ICG was coated with B16 cell membranes (I@BM NVs) by sonication and extrusion, and the morphological characteristics of I@BM NVs were evaluated by transmission electron microscopy (TEM) and NP-tracking analysis. Homologous cellular uptake was evaluated by flow cytometry (FCM) after staining by DiD dye. Cellular cytotoxicity was evaluated by cell counting kit-8 assay and the anti-tumor effect in vitro was assessed by FCM and western blotting. The anti-tumor effect in vivo was evaluated in a B16 xenograft model in mice. The tumor micro-environment was investigated by FCM and real-time PCR. Results: The vesicles are stable and uniform in nature, and show strong homologous targeting in vivo and in vitro. The vesicles can generate reactive oxygen species to induce apoptosis of B16 cells under near-infrared irradiation. Furthermore, the I@BM NVs induce a significant anti-tumor response in vivo, and perform better with respect to both tumor growth inhibition and lifespan extension. Analysis of immunocytes in the tumor microenvironment showed significant reductions in numbers of myeloid-derived suppressor cells and tumor-associated M2 macrophages in mice in the I@BM NVs group. This was accompanied by significant increases in numbers of M1 macrophages and proliferative CD4+/CD8+ T cells. Expression levels of IFN-γ and IL-2 increased in the I@BM NVs group, while expression of TGF-ß and IL-10 decreased. Conclusion: The results show that the I@BM NVs are feasible drugs for the treatment of melanoma by inducing cell apoptosis under NIR and shifting the immunosuppressive tumor microenvironment in vivo.

Identification of oral precancerous and cancerous tissue by swept source optical coherence tomography.

BACKGROUND AND OBJECTIVES: Distinguishing cancer from precancerous lesions is critical and challenging in oral medicine. As a noninvasive method, optical coherence tomography (OCT) has the advantages of real-time, in vivo, and large-depth imaging. Texture information hidden in OCT images can provide an important auxiliary effect for improving diagnostic accuracy. The aim of this study is to explore a reliable and accurate OCT-based method for the screening and diagnosis of human oral diseases, especially oral cancer. MATERIALS AND METHODS: Fresh ex vivo oral tissues including normal mucosa, leukoplakia with epithelial hyperplasia (LEH), and oral squamous cell carcinoma (OSCC) were imaged intraoperatively by a homemade OCT system, and 58 texture features were extracted to create computational models of these tissues. A principal component analysis algorithm was employed to optimize the combination of texture feature vectors. The identification based on artificial neural network (ANN) was proposed and the sensitivity/specificity was calculated statistically to evaluate the classification performance. RESULTS: A total of 71 sites of three types of oral tissues were measured, and 5176 OCT images of three types of oral tissues were used in this study. The superior classification result based on ANN was obtained with an average accuracy of 98.17%. The sensitivity and specificity of normal mucosa, LEH, and OSCC are 98.17% / 98.38%, 93.81% / 98.54%, and 98.11% / 99.04%, respectively. CONCLUSION: It is demonstrated from the high accuracies, sensitivities, and specificities that texture-based analysis can be used to identify oral precancerous and cancerous tissue in OCT images, and it has the potential to help surgeons in diseases screening and diagnosis effectively.

Classification of oral salivary gland tumors based on texture features in optical coherence tomography images.

Currently, the diagnoses of oral diseases primarily depend on the visual recognition of experienced clinicians. It has been proven that automatic recognition based on images can support clinical decision-making by extracting and analyzing objective hidden information. In recent years, optical coherence tomography (OCT) has become a powerful optical imaging technique with the advantages of high resolution and non-invasion. In our study, a dataset composed of four kinds of oral salivary gland tumors (SGTs) was obtained from a homemade swept-source OCT, including two benign and two malignant tumors. Seventy-six texture features were extracted from OCT images to create computational models of diseases. It was demonstrated that the artificial neural network (ANN) based on principal component analysis (PCA) can obtain high diagnostic sensitivity and specificity (higher than 99%) for these four kinds of tumors. The classification accuracy of each tumor is larger than 99%. In addition, the performances of two classifiers (ANN and support vector machine) were quantitatively evaluated based on SGTs. It was proven that the texture features in OCT images provided objective information to classify oral tumors.

Fast DNA Vaccination Strategy Elicits a Stronger Immune Response Dependent on CD8+CD11c+ Cell Accumulation.

Conventional DNA vaccine strategies usually employ a regimen of immunizations at 2-week or longer intervals to induce effective memory cell-dependent immune responses. Clinical cancer treatment requires a faster immunization strategy to contend with tumor progression. In this study, a novel fast immunization strategy was established, wherein a DNA vaccine was intramuscularly administered on days 0, 2, and 5 in a murine lung cancer model. Effector cells peaked 7 to 10 days after the last vaccination. Compared with traditional 2-week-interval immunization strategies, antigen-specific cytolysis and INF-γ secretion were significantly enhanced under the fast vaccination approach. As a result, the rapidly administered DNA vaccine elicited stronger and more prompt antitumor effects. The probable underlying mechanism of fast immunization was the accumulation of CD8+CD11c+ antigen-presenting cells at the injection site, which enhanced subsequent antigen presentation. In conclusion, the fast DNA vaccination strategy shortened vaccination time to 5 days and elicited a stronger antitumor immune response.

Comparative Evaluation of 4-Dimensional Computed Tomography and 4-Dimensional Magnetic Resonance Imaging to Delineate the Target of Primary Liver Cancer.

Purpose: To evaluate the feasibility of 4-dimensional magnetic resonance imaging (4DMRI) in establishing the target of primary liver cancer in comparison with 4-dimensional computed tomography (4DCT). Methods and Materials: A total of 23 patients with primary liver cancer who received radiotherapy were selected, and 4DCT and T2w-4DMRI simulations were conducted to obtain 4DCT and T2w-4DMRI simulation images. The 4DCT and T2w-4DMRI data were sorted into 10 and 8 respiratory phase bins, respectively. The liver and gross tumor volumes (GTVs) were delineated in all images using programmed clinical workflows under tumor delineation guidelines. The internal organs at risk volumes (IRVs) and internal target volumes (ITVs) were the unions of all the phase livers and GTVs, respectively. Then, the artifacts, liver volume, GTV, and motion range in 4DCT and T2w-4DMRI were compared. Results: The mean GTV volume based on 4DMRI was 136.42 ± 231.27 cm3, which was 25.04 cm3 (15.5%) less than that of 4DCT (161.46 ± 280.29 cm3). The average volume of ITV determined by 4DMRI was 166.12 ± 270.43 cm3, which was 22.44 cm3 (11.9%) less than that determined by 4DCT (188.56 ± 307.57 cm3). Liver volume and IRV in 4DMRI increased by 4.0% and 6.6%, respectively, compared with 4DCT. The difference in tumor motion by T2w-4DMRI based on the centroid was greater than that of 4DCT in the L/R, A/P, and S/I directions, and the average displacement differences were 2.6, 2.8, and 6.9 mm, respectively. The severe artifacts in 4DCT were 47.8% (11/23) greater than in 4DMRI 17.4% (4/23). Conclusions: Compared with 4DCT, T2-weighted and navigator-triggered 4DMRI produces fewer artifacts and larger motion differences in hepatic intrafraction tumors, which is a feasible technique for primary liver cancer treatment planning.

Synthesis of Conformationally Liberated Yohimbine Analogues and Evaluation of Cytotoxic Activity.

A modular synthetic approach to strategically unique structural analogues of the alkaloid yohimbine is reported. The overall synthetic strategy couples the transition-metal-catalyzed decarboxylative allylation of 2,2-diphenylglycinate imino esters with a scandium triflate-mediated highly endo-selective intramolecular Diels-Alder (IMDA) cycloaddition to generate a small collection of de-rigidified yohimbine analogues lacking the ethylene linkage between the indole and decahydroisoquinoline units. One compound generated in this study contains an unprecedented pentacyclic urea core and appears to demonstrate increased cytotoxicity against the gastric cancer cell line SGC-7901 in comparison to a pancreatic cancer cell line (PATU-8988) and a normal human gastric mucosal cell line (GES-1).

Prognostic Value of ctDNA Mutation in Melanoma: A Meta-Analysis.

PURPOSE: Melanoma is the most aggressive form of skin cancer. Circulating tumor DNA (ctDNA) is a diagnostic and prognostic marker of melanoma. However, whether ctDNA mutations can independently predict survival remains controversial. This meta-analysis assessed the prognostic value of the presence or change in ctDNA mutations in melanoma patients. METHODS: We identified studies from the PubMed, EMBASE, Web of Science, and Cochrane databases. We estimated the combined hazard ratios (HRs) for overall survival (OS) and progression-free survival (PFS) using either fixed-effect or random-effect models based on heterogeneity. RESULTS: Sixteen studies including 1,781 patients were included. Both baseline and posttreatment detectable ctDNA were associated with poor OS (baseline detectable vs. undetectable, pooled HR = 1.97, 95% CI = 1.64-2.36, P < 0.00001; baseline undetectable vs. detectable, pooled HR = 0.19, 95% CI = 0.11-0.36, P < 0.00001; posttreatment detectable vs. undetectable, pooled HR = 2.36, 95% CI = 1.30-4.28, P=0.005). For PFS, baseline detectable ctDNA may be associated with adverse PFS (baseline detectable vs. undetectable, pooled HR = 1.41, 95% CI = 0.84-2.37, P=0.19; baseline undetectable vs. detectable, pooled HR = 0.43, 95% CI = 0.19-0.95, P=0.04) and baseline high ctDNA and increased ctDNA were significantly associated with adverse PFS (baseline high vs. low/undetectable, pooled HR = 3.29, 95% CI = 1.73-6.25, P=0.0003; increase vs. decrease, pooled HR = 4.48, 95% CI = 2.45-8.17, P < 0.00001). The baseline BRAFV600 ctDNA mutation-positive group was significantly associated with adverse OS compared with the baseline ctDNA-negative group (pooled HR = 1.90, 95% CI = 1.58-2.29, P < 0.00001). There were no significant differences in PFS between the baseline BRAFV600 ctDNA mutation-detectable group and the undetectable group (pooled HR = 1.02, 95% CI = 0.72-1.44, P=0.92). CONCLUSION: The presence or elevation of ctDNA mutation or BRAFV600 ctDNA mutation was significantly associated with worse prognosis in melanoma patients.

Identification of tumor mutation burden-related hub genes and the underlying mechanism in melanoma.

Background: Tumor mutation burden (TMB) has emerged as an important predictive factor for drug resistance in cancers; however, the specific mechanism underlying TMB function in melanoma remains elusive. Methods: Data on somatic mutations, RNA sequencing (RNA-seq), miRNA sequencing (miRNA-seq), and clinical characteristics for 472 melanoma patients were extracted from the TCGA cohort. RNA-seq data of melanoma cell lines were obtained from the Cancer Cell Line Encyclopedia, and sensitivity of cell lines to therapeutic agents is available in the Cancer Therapeutics Response Portal. TMB was calculated based on somatic mutation data. Differentially expressed gene analysis, weighted gene co-expression network analysis, protein-protein interaction networks, Minimal Common Oncology Data Elements, and survival analysis were leveraged to determine TMB-related hub genes. Competing endogenous RNA (ceRNA) networks were constructed to explore the molecular mechanisms underlying hub gene function. The influence of key genes on drug sensitivity was analyzed to investigate their clinical significance. Results: Elevated TMB levels were significantly correlated with improved survival outcomes. In addition, six tumor-infiltrating immune cells, including naive B cells, regulatory T cells, memory resting CD4 T cells, memory B cells, activated mast cells, and resting NK cells, were significantly overexpressed in the low-TMB group relative to the high-TMB group. Furthermore, we identified FLNC, NEXN, and TNNT3 as TMB-related hub genes, and constructed their ceRNA networks, including five miRNAs (has-miR-590-3p, has-miR-374b-5p, has-miR-3127-5p, has-miR-1913, and has-miR-1291) and 31 lncRNAs (FAM66C, MIAT, NR2F2AS1, etc.). Finally, we observed that TMB-related genes were associated with distinct therapeutic responses to AKT/mTOR pathway inhibitors. Conclusions: We identified three TMB-associated key genes, established their ceRNA networks, and investigated their influence on therapeutic responses, which could provide insights into future precision medicine.

Classification of Salivary Gland Tumors Based on Quantitative Optical Coherence Tomography.

BACKGROUND AND OBJECTIVES: Visual inspection is the primary diagnostic method for oral diseases, and its accuracy of diagnosis mainly depends on surgeons' experience. Histological examination is still the golden standard, but it is invasive and time-consuming. In order to address these issues, as a noninvasive imaging technique, optical coherence tomography (OCT) can differentiate oral tissue with advantages of real-time, in situ, and high resolution. The aim of this study is to explore optimal quantitative parameters in OCT images to distinguish different salivary gland tumors. STUDY DESIGN/MATERIALS AND METHODS: OCT images of four salivary gland tumors were obtained from 14 patients, including mucoepidermoid carcinoma (MC), adenoid cystic carcinoma (ACC), basal cell adenoma (BCA), and pleomorphic adenoma (PA). Two parameters of optical attenuation coefficient (OAC) and standard deviation (SD) along the depth of OCT signal were combined to create a computational model of classification, and sensitivity/specificity of classification was calculated statistically to evaluate their results. RESULTS: A total of 5,919 two-dimensional (2D) OCT images were used for quantitative analysis. The classification sensitivities of 89.6%, 95.0%, 89.5%, 97.8%, and specificities of 97.6%, 99.0%, 98.0%, 98.2%, respectively, were obtained for MC, ACC, BCA, and PA, with the thresholds of 3.6 mm-1 based on OAC and 0.22/0.18 based on SD. CONCLUSION: It was demonstrated that OAC and SD could be considered as important parameters in quantitative analysis of OCT images for salivary gland tissue characterization and intraoperative diagnosis. It is of great potential value in promoting the application of this method based on OCT in clinical practice. Lasers Surg. © 2020 Wiley Periodicals LLC.

Overexpression of MdARD4 Accelerates Fruit Ripening and Increases Cold Hardiness in Tomato.

Ethylene plays an important role in stress adaptation and fruit ripening. Acireductone dioxygenase (ARD) is pivotal for ethylene biosynthesis. However, the response of ARD to fruit ripening or cold stress is still unclear. In this study, we identified three members of Malus ARD family, and expression profile analysis revealed that the transcript level of MdARD4 was induced during apple fruit ripening and after apple plants were being treated with cold stress. To investigate its function in cold tolerance and fruit ripening, MdARD4 was ectopically expressed in Solanum lycopersicum cultivar 'Micro-Tom', which has been considered as an excellent model plant for the study of fruit ripening. At the cellular level, the MdARD protein expressed throughout Nicotiana benthamiana epidermal cells. Overexpression of MdARD4 in tomato demonstrated that MdARD4 regulates the ethylene and carotenoid signaling pathway, increases ethylene and carotenoid concentrations, and accelerates fruit ripening. Furthermore, MdARD4 increased the antioxidative ability and cold hardiness in tomato. To conclude, MdARD4 may potentially be used in apple breeding to accelerate fruit ripening and increase cold hardiness.