Fine-needle Aspiration Washout Precipitation Specimens: An Acceptable Supplement to Genetic Mutation Detection of Thyroid Nodules.

Objectives: Thyroid nodules are common in adults, but only some of them are malignant. Ultrasound-guided fine-needle aspiration (FNA) is widely applied as a reliable and minimally invasive technique for evaluating thyroid nodules. However, the scarcity of FNA biopsy specimens poses a challenge to molecular diagnosis. This study aimed to evaluate the feasibility of FNA washout precipitation specimens as an effective supplement to the thyroid genetic test. Methods: A total of 115 patients with thyroid nodules were enrolled in our study. The BRAF V600E mutation status was detected in all FNA washout precipitation specimens and biopsy formalin-fixed paraffin-embedded (FFPE) specimens using an amplification refractory mutation system PCR (ARMS-PCR). All patients underwent cytological diagnoses; 79 patients also underwent surgery for histopathological analysis. Results: All the 115 samples were successfully analyzed using both FNA washout precipitation and biopsy FFPE specimens. The results showed that the BRAF V600E status detected in 96 FNA washout precipitation specimens were consistent with that in FNA biopsy FFPE specimens, including 41 BRAF V600E positive and 55 BRAF V600E negative, achieving a concordance rate of 84.4% (kappa = 0.689). Furthermore, the BRAF V600E mutation status using FNA washout precipitation specimens provided a 100.0% positive predictive value for diagnosing papillary thyroid carcinoma in patients with The Bethesda system for reporting thyroid cytopathology (TBSRTC) V. Besides, the BRAF V600E mutation status was positive in 90.9% (10/11) FNA washout precipitation specimens from patients with capsule invasion, achieving a higher overall sensitivity of 100.0%, compared with 57.1% of FNA washout precipitation specimens from patients without capsule invasion. Conclusion: These results suggested that FNA washout precipitation specimens might be a valuable supplementary sample type for detecting the BRAF V600E mutation in patients with thyroid nodules, especially with thyroid capsule invasion.

Inhibition of protein FAK enhances 5-FU chemosensitivity to gastric carcinoma via p53 signaling pathways.

The small molecule drug 5-fluorouracil (5-FU) is widely used in the treatment for gastric cancer (GC), however, it exerts poor efficacy and is associated with acquired and intrinsic resistance. Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, plays a key role in adhesion, migration, and proliferation of gastric carcinoma cells, suggesting that this kinase may be a promising therapeutic target. Differentially expressed FAK in GC tissue was detected by RT-qPCR and TCGA database analysis. To investigate the biological functions of FAK, loss-of-function experiments were performed. CCK-8 assay, colony formation assay, flow cytometry, dual-luciferase reporter assays, and western blot assays were conducted to determine the underlying mechanisms of FAK in 5-FU chemosensitivity in GC. FAK is overexpressed in GC patients, and positively correlated with poor prognosis. The use of shRNA interference to target FAK decreased proliferation and increased apoptosis of GC cells in vitro. Importantly, FAK silencing enhanced the therapeutic efficacy of 5-FU, leading to reduced tumor growth in vivo. We further demonstrated that FAK silencing increased 5-FU-induced caspase-3 activity, and promoted p53 transcriptional activities. Clinical data also has shown that patients with higher levels of FAK had significantly shorter overall survival (OS) and time to first progression (FP) than those with lower levels of FAK. These findings indicate that FAK plays a critical role in 5-FU chemosensitivity in GC, and the use of FAK inhibitors as an adjunct to 5-FU might be an effective strategy for patients who undergo chemotherapy.

Tannic acid inhibits NLRP3 inflammasome-mediated IL-1ß production via blocking NF-κB signaling in macrophages.

The NLRP3 inflammasome rapidly responds to many infections and stress signals and is involved in the pathogenesis of numerous inflammatory disease processes. Tannic acid plays a role in antioxidant, antifungal and antitumor activities. Here, we reported that tannic acid inhibited NLRP3 inflammasome activation by blocking NF-κB signaling to suppress IL-1ß secretion. We found that the BMDMs (bone marrow-derived macrophages cells) pre-treated with tannic acid blocked caspase-1 cleavage and inhibited IL-1ß secretion in a NLRP3-dependent manner, and suppressed NF-κB signaling activation by inhibiting NF-κB/P65 nuclear localization, suggesting that tannic acid inhibited NLRP3 inflammasome activation. These investigations revealed that tannic acid inhibited NLRP3 inflammasome activation via blocking NF-κB signaling in macrophages, providing us with evidence that tannic acid may be a potent inhibitor for NLRP3-driven diseases.

Hyphomonas beringensis sp. nov. and Hyphomonas chukchiensis sp. nov., isolated from surface seawater of the Bering Sea and Chukchi Sea.

Two Gram-negative, non-spore-forming, oval to pear shaped motile strains, designated 25B14_1(T) and BH-BN04-4(T), isolated from surface seawater from the Bering Sea and Chukchi Sea, respectively, were subjected to polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that strains 25B14_1(T) and BH-BN04-4(T) clustered together with Hyphomonas atlanticus 22II1-22F38(T) and Hyphomonas oceanitis DSM 5155(T), respectively, within genus Hyphomonas. Based on whole genome sequence analysis, the calculated DDH and ANIm values between strain 25B14_1(T) and BH-BN04-4(T) are 18.8 and 83.19% respectively. The calculated DDH values of strain 25B14_1(T) and BH-BN04-4(T) with seven type strains ranged from 18.2 to 19.9% and from 18.4 to 40.4%, respectively. The ANIm values of strain 25B14_1(T) and BH-BN04-4(T) with seven type strains ranged from 83.00 to 84.67% and from 83.14 to 90.58%, respectively. Both isolates were found to contain Q-11 as the predominant respiratory quinone. The major fatty acids of strain 25B14_1(T) were identified as C(16:0), C(17:0), C(18:1)ω7c-methyl and Summed Feature 8 (C(18:1)ω6c/ω7c as defined by MIDI), while in the case of strain BH-BN04-4(T) they were identified as C(16:0), C(18:1)ω7c-methyl and Summed Feature 8 (C(18:1)ω6c/ω7c). The G+C contents of 25B14_1(T) and BH-BN04-4(T) were determined to be 58.4 and 61.0 mol%, respectively. The combined phenotypic and genotypic data show that the two isolates each represent novel species of the genus Hyphomonas, for which the names Hyphomonas beringensis sp. nov. and Hyphomonas chukchiensis sp. nov. are proposed, with the type strain 25B14_1(T) (=MCCC 1A07321(T) = LMG 27914(T)) and BH-BN04-4(T) (=MCCC 1A07481(T) = LMG 27915(T)), respectively.

[Phylogenetic and degrading genes analysis of a PAH-degrading bacterium TVG9-VII from deep-sea hydrythermal environment].

OBJECTIVE: The aim of this study is to identify a polycyclic aromatic hydrocarbons (PAHs)-degrading bacterium isolated from deep-sea hydrothermal environment, including its taxonomy, characteristics and mechanism involved in PAH degradation. METHODS: The phylogeny was studied by 16S rRNA gene clone, and the degradation rates against different PAHs were determined by GC-MS. Meanwhile, PAH-degrading gene cluster was cloned by the genomic Fosmid library construction; the function of the key degrading-gene expression was examined by RT-PCR and qPCR to observe gene expression in the response to different PAHs. RESULTS: A PAH-degrading strain TVG9-VII was isolated from the hydrothermal chimney sample of the Lau basin in Southwest Pacific Ocean. It showed 99.7% similarities with 16S rRNA gene of Novosphingobium indicum strain H25(T). The degradation rates of this strain against phenanthrene, fluoranthene and pyrene were 95.2%, 57.3% and 69.6% in 21 days, respectively. A gene cluster, containing PAHs initial dioxygenase genes pheA1a and pheA1b, was obtained from genomic fosmid library, with the insertion size of 12.522 kb. The gene pheA1a was enhanced by 4.2 folds in mRNA expression in presence of phenanthrene, but expression enhancement was not observed in other tested PAHs including naphthalene, pyrene and fluoranthene. CONCLUSION: Strain TVG9-VII is isolated from deep-sea hydrothermal environment in genus Novosphingobium. It can degrade many kinds of PAHs, especially the high-weight-molecular PAHs.