Suppression of N-Glycosylation of Zinc Finger Protein 471 Affects Proliferation, Invasion, and Docetaxel Sensitivity of Tongue Squamous Cell Carcinoma via Regulation of c-Myc.

Zinc finger protein 471 (ZNF471) is a member of the Krüppel-related domain zinc finger protein family, and has recently attracted attention because of its anti-cancer effects. N-glycosylation regulates expression and functions of the protein. This study aimed to investigate the effects of ZNF471 N-glycosylation on the proliferation, invasion, and docetaxel sensitivity of tongue squamous cell carcinoma (TSCC). It analyzed the expression, function, and prognostic significance of ZNF471 in TSCC using bioinformatics techniques such as gene differential expression analysis, univariate Cox regression analysis, functional enrichment analysis, and gene set enrichment analysis. Using site-specific mutagenesis, this study generated three mutant sites for ZNF471 N-glycosylation to determine the effect of N-glycosylation on ZNF471 protein levels and function. Quantitative real-time PCR, Western blot analysis, and immunohistochemistry tests confirmed the down-regulation of ZNF471 expression in TSCC. Low expression of ZNF471 is associated with poor prognosis of patients with TSCC. Overexpression of ZNF471 in vitro retarded the proliferation of TSCC cells and suppressed cell invasion and migration ability. Asparagine 358 was identified as a N-glycosylation site of ZNF471. Suppressing N-glycosylation of ZNF471 enhanced the protein stability and promoted the translocation of protein to the cell nucleus. ZNF471 binding to c-Myc gene promoter suppressed oncogene c-Myc expression, thereby playing the anti-cancer effect and enhancing TSCC sensitivity to docetaxel. In all, N-glycosylation of ZNF471 affects the proliferation, invasion, and docetaxel sensitivity of TSCC via regulation of c-Myc.

Low-Frequency Evolution Mechanism of Customized HEAs-Based Electromagnetic Response Modes Manipulated by Carbothermal Shock.

An emerging carbothermal shock method is an ultra-convenient strategy for synthesizing high-entropy alloys (HEAs), in which the intelligent combination of carbon support and HEAs can be serve as a decisive factor for interpreting the trade-off relationship between conductive gene and dielectric gene. However, the feedback mechanism of HEAs ordering degree on electromagnetic (EM) response in 2-18 GHz has not been comprehensively demystified. Herein, while lignin-based carbon fiber paper (L-CFP) as carbon support, L-CFP/FeCoNiCuZn-X with is prepared by carbothermal shock method. The reflection loss of -82.6 dB with thickness of 1.31 mm is achieved by means of pointing electron enrichment within L-CFP/FeCoNiCuZn HEAs heterointerfaces verified by theoretical calculations. Simultaneously, low-frequency evolution with high-intensity and broadband EM response relies on a "sacrificing" strategy achieved by construction of polymorphic L-CFP/semi-disordered-HEAs heterointerfaces. The practicality of L-CFP/FeCoNiCuZn-X in complex environments is given prominence to thermal conductivity, hydrophobicity, and electrocatalytic property. This work is of great significance for insightful mechanism analysis of HEAs in the application of electromagnetic wave absorption.

Cognitive behavior therapy for depression in people with epilepsy: A systematic review and meta-analysis.

BACKGROUND: Cognitive behavioral therapy (CBT) is the recommended treatment for depression in patients with epilepsy (PWE). However, there are no studies that calculate the effect size of CBT on depression and quality of life (QoL) in PWE. METHODS: We searched seven electronic databases (PubMed, Web of Science, Embase, Cochrane Library, Clinical Trials, Ovid Medline, and PsycINFO). We included 13 studies examining CBT for depression in PWE and calculated its effect size. RESULTS: A total of 13 studies met the criteria. After treatment, CBT improves depression in PWE (g = 0.36, 95%CI: 0.18 to 0.54, I2 = 50%), and the efficacy maintains during follow-up (g = 0.47, 95%CI: 0.04 to 0.89, I2 = 80%). Subgroup analysis has shown that individual CBT (g = 0.47, 95%CI: 0.20 to 0.73, I2 = 0%) had a greater effect size than group CBT (g = 0.30, 95%CI: 0.07 to 0.53, I2 = 62%) in the treatment of depression. Likewise, CBT has a positive effect on the QoL improvement of PWE (g = 0.34, 95%CI: 0.11 to 0.57, I2 = 64%). In controlling seizures, CBT did not differ from the control group (g = -0.06, 95%CI: -0.32 to 0.19, I2 = 0%). CONCLUSIONS: Cognitive behavioral therapy interventions were effective in improving depression and QoL in PWE, but not effective in controlling seizures. The efficacy of CBT interventions targeting seizure control seems to be uncertain.

Clinicopathological and prognostic significance of PD-1/PD-L1 axis expression in patients with tongue squamous cell carcinoma.

Tongue squamous cell carcinoma (TSCC) is more aggressive than other head and neck tumors, and the prognosis for patients with advanced TSCC is poor. At present, comprehensive treatment based on surgery as the main method is not effective for patients with advanced TSCC. The application of PD-1/PD-L1 immunocheckpoint inhibitor alone in patients with TSCC has not been reported. To explore the role of PD-1/PD-L1, we investigated the expression of PD-1 and PD-L1 in TSCC and analyzed the relationship between the expression of PD-1 and PD-L1 and the related clinicopathological parameters and survival prognosis. The expression of PD-1 was significantly associated with palindromia (p = .015) and maximum diameter (p = .043). The expression of PD-L1 in tumor cells was significantly associated with N stage (P = .024), chemotherapy (p = .032), and clinical stage (p = .019). The expression of PD-L1 in infiltrating lymphocytes was significantly associated with palindromia (p = .030). Univariate and multivariate Cox analyses for prognoses of patients showed significant prognostic factors of overall survival and relapse-free survival. The high expression of PD-L1 on infiltrating lymphocytes for OS and RFS was an independent protective factor for patients with TSCC. The high expression of PD-1 on infiltrating lymphocytes and clinical stage for OS and RFS were independent risk factors for patients with TSCC. The data provide a reference for clinical treatment of TSCC with immunotherapy.

Exome sequencing identifies a novel CEACAM16 mutation associated with autosomal dominant nonsyndromic hearing loss DFNA4B in a Chinese family.

Autosomal dominant nonsyndromic hearing loss (ADNSHL/DFNA) is a highly genetically heterogeneous disorder. Hitherto only about 30 ADNSHL-causing genes have been identified and many unknown genes remain to be discovered. In this research, genome-wide linkage analysis mapped the disease locus to a 4.3 Mb region on chromosome 19q13 in SY-026, a five-generation nonconsanguineous Chinese family affected by late-onset and progressive ADNSHL. This linkage region showed partial overlap with the previously reported DFNA4. Simultaneously, probands were analyzed using exome capture followed by next-generation sequencing. Encouragingly, a heterozygous missense mutation, c.505G>A (p.G169R) in exon 3 of the CEACAM16 gene (carcinoembryonic antigen-related cell adhesion molecule 16), was identified via this combined strategy. Sanger sequencing verified that the mutation co-segregated with hearing loss in the family and that it was not present in 200 unrelated control subjects with matched ancestry. This is the second report in the literature of a family with ADNSHL caused by CEACAM16 mutation. Immunofluorescence staining and western blots also prove CEACAM16 to be a secreted protein. Furthermore, our studies in transfected HEK293T cells show that the secretion efficacy of the mutant CEACAM16 is much lower than that of the wild type, suggesting a deleterious effect of the sequence variant.

Baicalein induces human osteosarcoma cell line MG-63 apoptosis via ROS-induced BNIP3 expression.

Baicalein, a flavonoid compound, is one of the active constituents of the root of Scutellariae Radix. Its antitumor effects have attracted widespread attention worldwide. One of its major functions is to induce the apoptosis of tumor cells, but the antitumor mechanism is currently unclear. In the present study, we found that baicalein increased MG-63 cell mortality in a dose-dependent manner. Meanwhile, baicalein activated apoptosis through induced intracellular reactive oxygen species (ROS) generation, and that ROS scavenger N-acetyl-cysteine (NAC), glutathione (GSH), and superoxide dismutase (SOD) apparently inhibited intracellular ROS production, consequently attenuating the baicalein-induced apoptosis. Baicalein also induce the mitochondrial fragmentation which precedes the cell apoptosis. This morphological alteration is accompanied by an increase in the expression of the protein BNIP3 as well as Mul1 and Drp1. Furthermore, we show that the inhibition of BNIP3 expression can inhibit cell apoptosis by baicalein treatment. Taken together, our results bring the evidence of a mechanism that links apoptosis and ROS-induced BNIP3 expression in MG-63 cells with bacalein treatment and suggest that baicalein has a good potential as an anti-osteosarcoma drug.

Spontaneous Orientation Polarization of Anisotropic Equivalent Dipoles Harnessed by Entropy Engineering for Ultra-Thin Electromagnetic Wave Absorber.

The synthesis of carbon supporter/nanoscale high-entropy alloys (HEAs) electromagnetic response composites by carbothermal shock method has been identified as an advanced strategy for the collaborative competition engineering of conductive/dielectric genes. Electron migration modes within HEAs as manipulated by the electronegativity, valence electron configurations and molar proportions of constituent elements determine the steady state and efficiency of equivalent dipoles. Herein, enlightened by skin-like effect, a reformative carbothermal shock method using carbonized cellulose paper (CCP) as carbon supporter is used to preserve the oxygen-containing functional groups (O·) of carbonized cellulose fibers (CCF). Nucleation of HEAs and construction of emblematic shell-core CCF/HEAs heterointerfaces are inextricably linked to carbon metabolism induced by O·. Meanwhile, the electron migration mode of switchable electron-rich sites promotes the orientation polarization of anisotropic equivalent dipoles. By virtue of the reinforcement strategy, CCP/HEAs composite prepared by 35% molar ratio of Mn element (CCP/HEAs-Mn2.15) achieves efficient electromagnetic wave (EMW) absorption of - 51.35 dB at an ultra-thin thickness of 1.03 mm. The mechanisms of the resulting dielectric properties of HEAs-based EMW absorbing materials are elucidated by combining theoretical calculations with experimental characterizations, which provide theoretical bases and feasible strategies for the simulation and practical application of electromagnetic functional devices (e.g., ultra-wideband bandpass filter).

Cellular metabolism: A key player in cancer ferroptosis.

Cellular metabolism is the fundamental process by which cells maintain growth and self-renewal. It produces energy, furnishes raw materials, and intermediates for biomolecule synthesis, and modulates enzyme activity to sustain normal cellular functions. Cellular metabolism is the foundation of cellular life processes and plays a regulatory role in various biological functions, including programmed cell death. Ferroptosis is a recently discovered form of iron-dependent programmed cell death. The inhibition of ferroptosis plays a crucial role in tumorigenesis and tumor progression. However, the role of cellular metabolism, particularly glucose and amino acid metabolism, in cancer ferroptosis is not well understood. Here, we reviewed glucose, lipid, amino acid, iron and selenium metabolism involvement in cancer cell ferroptosis to elucidate the impact of different metabolic pathways on this process. Additionally, we provided a detailed overview of agents used to induce cancer ferroptosis. We explained that the metabolism of tumor cells plays a crucial role in maintaining intracellular redox homeostasis and that disrupting the normal metabolic processes in these cells renders them more susceptible to iron-induced cell death, resulting in enhanced tumor cell killing. The combination of ferroptosis inducers and cellular metabolism inhibitors may be a novel approach to future cancer therapy and an important strategy to advance the development of treatments.

Novel multiplex primer extension and denaturing high-performance liquid chromatography for genotyping of the deafness gene mutations.

OBJECTIVE: To find a rapid and accurate genotyping method for specific non-syndromic hearing loss (NSHL)-causing gene mutations for disease diagnosis in different ethnic populations. METHODS: We performed a novel multiplex primer extension (PE) reaction in combination with denaturing high-performance liquid chromatography (DHPLC) to simultaneously detect and genotype the 6 most common mutations in 180 patients with NSHL (GJB2-235delC, GJB2-299delAT, PDS-A2168G, PDS IVS7-2A>G, mtDNA-A1555G, and mtDNA-C1494T) in Chinese population. This method involved the amplification of the target sequence, followed by a purification step, a multiplex PE reaction, and DHPLC analysis performed on the Transgenomic Wave DNA fragment analysis system under fully-denaturing conditions. RESULTS: In a blind analysis, this technique successfully and accurately genotyped 100% of the samples simultaneously characterized by direct sequencing. CONCLUSION: Combination of PE and DHPLC is simple, rapid, accurate, and cost-effective for genotyping common disease-causing mutations, including substitutions, insertions, and deletions in NSHL, and may be successfully used in other genetic diseases.

High Efficiency Gas Permeability Membranes from Ethyl Cellulose Grafted with Ionic Liquids.

Ethyl cellulose was grafted with ionic liquids in optimal yields (62.5-64.1%) and grafting degrees (5.93-7.90%) by the esterification of the hydroxyl groups in ethyl cellulose with the carboxyl groups in ionic liquids. In IR spectra of the ethyl cellulose derivatives exhibited C=O bond stretching vibration peaks at 1760 or 1740 cm-1, confirming the formation of the ester groups and furnishing the evidence of the successful grafting of ethyl cellulose with ionic liquids. The ethyl cellulose grafted with ionic liquids could be formed into membranes by using the casting solution method. The resulting membranes exhibited good membrane forming ability and mechanical properties. The EC grafted with ionic liquids-based membranes demonstrated PCO2/PCH4 separation factors of up to 18.8, whereas the PCO2/PCH4 separation factor of 9.0 was obtained for pure EC membrane (both for CO2/CH4 mixture gas). The membranes also demonstrated an excellent gas permeability coefficient PCO2, up to 199 Barrer, which was higher than pure EC (PCO2 = 46.8 Barrer). Therefore, it can be concluded that the ionic liquids with imidazole groups are immensely useful for improving the gas separation performances of EC membranes.

Loss of miR-16 contributes to tumor progression by activation of tousled-like kinase 1 in oral squamous cell carcinoma.

A different expression signature of miRNA in oral squamous cell carcinoma (OSCC) has been validated. MicroRNA-16 (miR-16) as one of the distinctly dysregulated miRNAs in OSCC, its functional role in progression of OSCC remains not fully clear. Herein, miR-16 expression was significantly lower in OSCC tissues compared to that in adjacent normal tissues (n = 131). A lower level of miR-16 was found to be associated with poor prognosis on a cohort of 131 patients with OSCC, and on an extensive public data (457) from TCGA database. Additionally, expression of TLK1 was significantly higher in OSCC tissues compared to that in adjacent normal tissues, which is negatively correlated with miR-16 expression in OSCC. Bioinformatics analyses exhibited that TLK1 is a potential downstream effector of miR-16 by directly targeting the 3'-untranslated regions (3'-UTR) of mRNA. Forced expression of miR-16 in OSCC cell lines inhibits cell proliferation in vitro, and tumor growth in vivo by inhibition of TLK1. Mechanistically, downregulation of TLK1 by miR-16 enhances higher level of DNA damage leading to a significant increase of G2/M arrest in SCC9 cells. And, overexpression of TLK1 substantially reduces DNA damage and G2/M arrest by activation of TLK1-dependent cell cycle checkpoint response. To conclude, miR-16 is downregulated in OSCC and serves as tumor suppressor in OSCC progression by targeting TLK1, which has potential to be the novel therapeutic targets and diagnostic biomarkers for OSCC.

[Application of free anterolateral thigh Kiss flap in repair of large scalp defect after malignant tumor resection].

Objective: To investigate the effectiveness of free anterolateral thigh Kiss flap in repair of large scalp defect after malignant tumor resection. Methods: Between December 2012 and December 2016，18 patients with large scalp defect after malignant tumor resection were treated. There were 16 males and 2 females with an average age of 52.6 years (range, 43-62 years). There were 17 cases of squamous carcinoma and 1 case of dermatofibrilsarcoma protuberan. The size of scalp defect ranged from 15 cm×10 cm to 17 cm×12 cm after resection of tumors. The scalp defects were repaired with the free anterolateral thigh Kiss flap. And the size of flap ranged from 15 cm×6 cm to 20 cm×8 cm. The skull was completely resected in 2 cases, and repaired with Titanium mesh. The sizes of skull defects were 12 cm×10 cm and 10 cm×8 cm. The donor site was sutured directly. Results: Eighteen flaps survived with primary healing of wounds; and healing by first intention was obtained at the donor sites. One patient died because of intracranial metastasis at 5 months after operation, and no local recurrence occurred in the other 17 patients. The follow-up time ranged from 6 months to 4 years (mean, 26.6 months). The results of both appearance and function were satisfactory, without ulceration during follow-up. No obvious scar was found at donor sites and no obvious impairment was observed after harvesting free anterolateral thigh flap. Conclusion: Large scalp defects after malignant tumor resection can be effectively repaired by free anterolateral thigh Kiss flap. The donor site can be sutured directly, without skin grafting, thus avoiding the secondary donor site.

Digenic mutations involving both the BSND and GJB2 genes detected in Bartter syndrome type IV.

Bartter syndrome type IV, characterized by salt-losing nephropathies and sensorineural deafness, is caused by mutations of BSND or simultaneous mutations of both CLCNKA and CLCNKB. GJB2 is the primary causative gene for non-syndromic sensorineural deafness and associated with several syndromic sensorineural deafness. Owing to the rarity of Bartter syndrome, only a few mutations have been reported in the abovementioned causative genes. To investigate the underlying mutations in a Chinese patient with Bartter syndrome type IV, genetic analysis of BSND, CLCNKA, CLCNKB and GJB2 were performed by polymerase chain reaction and direct sequencing. Finally, double homozygous mutations c.22C > T (p.Arg8Trp) and c.127G > A (Val43Ile) were detected in exon 1 of BSND. Intriguingly, compound heterozygous mutations c.235delC (p.Leu79CysfsX3) and c.109G > A (p.Val37Ile) were also revealed in exon 2 of GJB2 in the same patient. No pathogenic mutations were found in CLCNKA and CLCNKB. Our results indicated that the homozygous mutation c.22C > T was the key genetic reason for the proband, and a digenic effect of BSND and GJB2 might contributed to sensorineural deafness. To our knowledge, it was the first report showing that the GJB2 gene mutations were detected in Bartter syndrome.

Identification and functional analysis of a novel mutation in the SOX10 gene associated with Waardenburg syndrome type IV.

Waardenburg syndrome type IV (WS4) is a rare genetic disorder, characterized by auditory-pigmentary abnormalities and Hirschsprung disease. Mutations of the EDNRB gene, EDN3 gene, or SOX10 gene are responsible for WS4. In the present study, we reported a case of a Chinese patient with clinical features of WS4. In addition, the three genes mentioned above were sequenced in order to identify whether mutations are responsible for the case. We revealed a novel nonsense mutation, c.1063C>T (p.Q355*), in the last coding exon of SOX10. The same mutation was not found in three unaffected family members or 100 unrelated controls. Then, the function and mechanism of the mutation were investigated in vitro. We found both wild-type (WT) and mutant SOX10 p.Q355* were detected at the expected size and their expression levels are equivalent. The mutant protein also localized in the nucleus and retained the DNA-binding activity as WT counterpart; however, it lost its transactivation capability on the MITF promoter and acted as a dominant-negative repressor impairing function of the WT SOX10.

Osteopontin expression in nasopharyngeal carcinoma: its relevance to the clinical stage of the disease.

PURPOSE: To investigate osteopontin (OPN) expression in human nasopharyngeal carcinoma (NPC) and evaluate its clinical significance in the disease. MATERIALS AND METHODS: The expression of OPN mRNA in 44 frozen NPC tissue and 15 normal nasopharyngeal epithelium tissue (NNET) samples was examined by semi-quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). OPN protein expression in 67 paraffin-embedded NPC tissue and 21 NNET samples was detected by immunohistochemistry (IHC). In addition, OPN expression was investigated in 12 paired NPC and para-carcinoma tissue (PCT) samples by western blotting (WB). The association between the expression of OPN and the clinicopathologic parameters of NPC was evaluated. RESULTS: Three different methods all showed that the expression of OPN mRNA or protein in NPC was significantly higher than in NNET or PCT (P = 0.000, 0.001, 0.000, respectively). After an examination by IHC, 88.1% (59/67) of NPC samples showed strong or moderate positive OPN staining and 28.6% (6/21) of NNET samples displayed a weak positive OPN staining. The staining of OPN in tumor cells was mainly localized to the cytoplasm. OPN expression in NPC was not related to patient age or sex (P > 0.05), but was significantly related to tumor size, regional lymph nodal metastasis, and NPC clinical stages (P < 0.05). CONCLUSIONS: Our study demonstrated that OPN mRNA and protein overexpression in NPC may be important in the pathogenesis of the disease. It was strongly related to T stage, N stage and clinical stages of NPC, suggesting that OPN may be involved in NPC metastasis and progression.

Unique penetrance of hearing loss in a five-generation Chinese family with the mitochondrial 12S rRNA 1555A > G mutation.

CONCLUSIONS: Analysis of the complete mtDNA genome and X-linkage of this five-generation Chinese family revealed that the 1555A > G mutation may lead to deafness. OBJECTIVES: Mutations in mitochondrial DNA (mtDNA) have been found to be associated with sensorineural hearing loss. However, the variable clinical phenotype and incomplete penetrance of mtDNA 1555A > G-induced hearing loss complicate our understanding of this mutation. We aimed to identify whether nuclear genes, mitochondrial haplotypes/variants, and a possible threshold effect are involved in its manifestation in the pedigree. METHODS: We performed clinical, genetic, and X-linkage analysis of a five-generation Chinese family in which all the affected individuals were male. RESULTS: Clinical evaluation revealed that affected individuals with or without aminoglycoside exposure developed hearing loss extending gradually from 8000 Hz to 4000 Hz and then to 1000 Hz. Using X-linkage analysis and sequencing, we detected an identical homoplasmic 1555A > G mutation in nine individuals, and a previously unreported variant 14163C > T in mtDNA. The new variant 14163C > T coexisted with the 1555A > G mutation in six affected subjects of our pedigree. The previously unreported variant 14163C > T and aminoglycoside exposure may synergize the development of this deafness.

Identification of Galectin-1 as a novel biomarker in nasopharyngeal carcinoma by proteomic analysis.

Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China and Southeast Asia. It is characterized as a multistep process involved in multiple genetic and epigenetic events. The mechanism of carcinogenesis still needs to be further clarified. In this study, two-dimensional gel electrophoresis, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), RT-PCR, Western blot and immunohistochemical (IHC) analyses were used to detect Galectin-1 expression in NPC compared with normal nasopharyngeal epithelial tissues (NNET). We found that Galectin-1 was expressed at a significantly higher level in NPC compared with NNET. Our results indicated that high expression level of Galectin-1 might correlate with the development of NPC and Galectin-1 may serve as a potential diagnostic marker or therapeutic target for NPC.