METTL3 modulates m6A modification of CDC25B and promotes head and neck squamous cell carcinoma malignant progression.

BACKGROUND: N6-methyladenosine (m6A) RNA methylation and its methyltransferase METTL3 have been widely reported to be involved in different cancers by regulating RNA metabolism and function. Here, we aimed to explore the biological function and clinical significance of m6A modification and METTL3 in head and neck squamous cell carcinoma (HNSCC). METHODS: The prognostic value of METTL3 expression was evaluated using tissue microarray and immunohistochemical staining analyses in a human HNSCC cohort. The biological role and mechanism of METTL3 in HNSCC tumour growth, metastasis and angiogenesis were determined in vitro and in vivo. RESULTS: M6A levels and METTL3 expressions in HNSCC tissues were significantly increased compared with paired adjacent tissues. Meanwhile, METTL3 was an independent risk factor for the prognosis of HNSCC patients. Moreover, METTL3 overexpression promoted HNSCC cell proliferation, migration, invasion, and angiogenesis, while knockdown of METTL3 had an opposite effect in vivo and in vitro. Mechanistically, METTL3 enhanced the m6A modification of CDC25B mRNA, which maintained its stability and upregulated its expression, thereby activating G2/M phase of cell cycle and leading to HNSCC malignant progression. CONCLUSIONS: METTL3 may be a potential prognostic biomarker and therapeutic target for HNSCC.

The correlation between FeNO and nNO in allergic rhinitis and bronchial asthma.

ABSTRACT: This study aimed to evaluate the correlation between fractional exhaled nitric oxide (FeNO) and nasal nitric oxide (nNO) in allergic rhinitis (AR) and patients with or without bronchial asthma (BA).A total of 90 patients who were diagnosed with persistent AR (AR group, n = 30), BA (BA group, n = 30), or allergic rhinitis with bronchial asthma (AR-BA) (AR-BA group, n = 30), were enrolled in this study, along with 30 healthy adult volunteers (control group, n = 30). The participants were further divided into 2 groups based on the results of a skin-prick test (SPT): a highly atopic group (SPT = 3+ and above) and a moderately atopic group (SPT = 2+ and below). All participants underwent FeNO and nNO measurement, an absolute blood eosinophil count, total serum immunoglobulin measurement, and horizontal baseline lung capacity determination.The results showed that the FeNO levels in the 3 observation groups were significantly higher than those in the control group (P < .01), and in the BA group they were significantly higher than in the AR-BA group (P < .01). The levels of nNO in both the AR group and the AR-BA group were higher than those in the control group and the BA group (P < .01), but there was no significant difference between the AR group and the AR-BA group (P > .05). The levels of nNO in the BA group were also significantly different from those in the control group (P < .01).FeNO and nNO are positively correlated with the degree of AR in patients with BA; therefore, nNO levels can be used as an inflammatory marker of AR in patients with BA. FeNO can also be used as an inflammatory marker of AR in patients complicated with BA as a warning indicator of asthma.

Carbon monoxide-releasing molecule-3 protects against cortical pyroptosis induced by hemorrhagic shock and resuscitation via mitochondrial regulation.

OBJECTIVE: Carbon monoxide (CO) releasing molecule (CORM)-3, a water-soluble CORM, has protective effects against inflammatory and ischemia/reperfusion injury. We determined the effect of CORM-3 against neuronal pyroptosis in a model of hemorrhagic shock and resuscitation (HSR) in rats via mitochondrial regulation. METHODS: Rats were treated with CORM-3 (4 mg/kg) in vitro after HSR. We measured cortical CO content 3-24 h after HSR; assessed neuronal pyroptosis, mitochondrial morphology, ROS production, and mitochondrial membrane potential at 12 h after HSR; and evaluated brain magnetic resonance imaging at 24 h after HSR and learning ability 30 days after HSR. We also measured soluble guanylate-cyclase (sGC)-cyclic guanosine monophosphate (cGMP) signaling pathway activity using a blocker of sGC, NS2028, and 125I-cGMP assay. RESULTS: Among rats that underwent HSR, CORM-3-treated rats had more CO in the cortical tissue than sham- and iCORM-3-treated rats. CORM-3-treated rats had significantly less neuronal pyroptosis in the cortical tissue; higher sGC activity and cGMP content; lower ROS production; better mitochondrial morphology, function, and membrane potential; and enhanced learning/memory ability than HSR-treated rats. However, these neuroprotective effects of CORM-3 were partially inhibited by NS2028. CONCLUSION: CORM-3 may alleviate neuronal pyroptosis and improve neurological recovery in HSR through mitochondrial regulation mediated by the sGC-cGMP pathway. Thus, CO administration could be a promising therapeutic strategy for hemorrhagic shock.

2,4-Disubstituted quinazolines targeting breast cancer cells via EGFR-PI3K.

A series of novel 2,4-disubstituted quinazolines were synthesized and evaluated for their anti-tumor activity against five human cancer cells (MDA-MB-231, MCF-7, PC-3, HGC-27 and MGC-803) using MTT assay. Among them, compound 9n showed the most potent cytotoxicity against breast cancer cells. Compound 9n also significantly inhibited the colony formation and migration of MDA-MB-231 and MCF-7 cells. Meanwhile, compound 9n induced cell cycle arrest at G1 phase and cell apoptosis, as well as increased accumulation of intracellular ROS. Furthermore, compound 9n exerted anti-tumor effects in vitro via decreasing the expression of anti-apoptotic protein Bcl-2 and increasing the pro-apoptotic protein Bax and p53. Mechanistically, compound 9n markedly decreased p-EGFR and p-PI3K expression, which revealed that compound 9n targeted breast cancer cells via interfering with EGFR-PI3K signaling pathway. Molecular docking suggested that compound 9n could indeed bind into the active pocket of EGFR. All the findings suggest that compound 9n might be a valuable lead compound for anti-tumor agents targeting breast cancer cells.

Effects of gap junctions blockers on fast ripples and connexin in rat hippocampi after status epilepticus.

OBJECTIVE: To assess the dynamic changes in the average and peak spectral power of fast ripples (FRs) in the hippocampi after interventions with valproate sodium (VPA), carbenoxolone (CBX) and quinine (QUIN). METHOD: Adult rats were used to establish a lithium-pilocarpine (pilo) epileptic model, and were assigned to a lithium-pilocarpine (PILO), VPA + PILO, QUIN + PILO or CBX + PILO group. Intracranial electroencephalography was performed before and after status epilepticus (SE). The hippocampal connexin (CX) 43, CX32 and CX36 expressions were analyzed via western blotting. RESULTS: The time required for the disappearance of SE after chloral hydrate injection was lower in the intervention groups than in the PILO group (p < 0.05). Seizures induced CX43 expression, but had no significant effects on CX36 or CX32 expressions. Pretreatment with VPA, QUIN and CBX inhibited CX43, CX36 and CX32 expression after SE. The average spectral power of the FRs was significantly lower in the VPA + PILO and QUIN + PILO groups than in the PILO group at 10 min after SE, 10 min before chloral hydrate injection, and 10 min after chloral hydrate injection (p < 0.05). The average spectral power of FRs was lower in the CBX + PILO group than in the PILO group at 10 min after SE (p < 0.05). The average spectral power of FRs in the 3 intervention groups recovered to the baseline level at 10 min after chloral hydrate injection and persisted for 3 days after SE. The dynamic changes in the average and peak spectral power of FRs were similar. SIGNIFICANCE: After SE, CX may participate in pathological FR generation by establishing abnormal electrical synaptic transmission. Gap junction blockers can inhibit various CX expressions, and thus decrease FR energy and alleviate the degree of seizure. These findings could contribute to the development of new anti-epileptic drugs with novel mechanistic targets.

Role of epithelial-mesenchymal transition markers E-cadherin, N-cadherin, ß-catenin and ZEB2 in laryngeal squamous cell carcinoma.

Epithelial-mesenchymal transition (EMT) allows neoplastic cells to gain the invasive phenotype and become migratory, which is required for cancer progression and metastasis. In the present study, the expression of EMT-associated biomarkers and their association with clinicopathological parameters in laryngeal squamous cell carcinoma (LSCC) was investigated. E-cadherin, N-cadherin, ß-catenin and zinc finger E-box binding homeobox 2 (ZEB2) protein expression was evaluated with immunohistochemistry in a cohort of 76 patients with operable LSCC. The association between these transition markers, clinicopathological parameters and their prognostic impact in LSCC was analyzed. Immunohistochemical analysis revealed that EMT-associated proteins were differentially expressed between LSCC and adjacent non-neoplastic laryngeal tissue. Negative E-cadherin expression and positive N-cadherin, ß-catenin and ZEB2 expression were associated with a later tumor (T) stage, decreasing tumor differentiation and a reduced overall survival (OS) time (OS: E-cadherin, P=0.016; N-cadherin, P=0.003; ß-catenin, P=0.002; ZEB2, P=0.0003). E-cadherin/ß-catenin co-expression was significantly associated with the majority of clinicopathological parameters assessed, including lymph node metastases, T stage and tumor cell differentiation (P=0.004, P=0.005, and P<0.001, respectively). Multivariate analysis indicated that T stage and the positive expression of ß-catenin and ZEB2 were independent risk factors for OS in LSCC (P=0.014, P=0.025 and P=0.003, respectively). It was concluded that EMT mediates tumor progression, and reduces OS time in patients with LSCC. E-cadherin/ß-catenin co-expression may be associated with clinicopathological parameters. T stage, and the positive co-expression of ß-catenin and ZEB2 may be independent predictors of prognosis in LSCC.

Risk factors for poor prognosis in children with refractory purulent meningitis and the discharge criteria.

This study was undertaken to investigate the relationship between cerebrospinal fluid abnormalities and prognosis in pediatric refractory purulent meningitis. Ninety cases of pediatric refractory purulent meningitis were stratified into "good" (n=33) or "poor" (n=57) prognosis groups according to the Glasgow clinical outcome scores. The symptoms, laboratory results, and prognosis were compared by using univariate and multivariate logistic regression analyses. Univariate analysis showed that poor prognosis was associated with: unequal pupil size in both eyes; positive Babinski sign; CSF-WBC >500×106/L, CSF protein concentration >1.0g/L, CSF glucose content <1.5mmol/L; initial procalcitonin result >0.1ng/dL on admission; hemoglobin <90g/L during hospitalization; abnormal head imaging, and abnormal electroencephalogram. On multivariate analysis only unequal pupil size in both eyes and CSF glucose content <1.5mmol/L remained significant. The CSF protein concentration was significantly different between groups at discharge. The cutoff value was 0.68g/L. We recommend that discharged patients meet the following criteria: full antibiotic course and over 1 week of defervesce, disappearance of acute phase symptoms, CSF-WBC ≤28×106/L, CSF glucose >1.75mmol/L, and protein <0.68g/L. The patient may be discharged for follow-up if no relapse occurs during 3-5 days of observation after drug withdrawal.

The Efficacy and Tolerability of the Clonidine Transdermal Patch in the Treatment for Children with Tic Disorders: A Prospective, Open, Single-Group, Self-Controlled Study.

BACKGROUND: To evaluate the efficacy and tolerability of a clonidine transdermal patch in the treatment of children with tic disorders (TD) and to establish a predictive model for patients. METHODS: Forty-one patients who met the inclusion criteria entered into 12 weeks of prospective, open, single-group, self-controlled treatment with a clonidine transdermal patch. The Yale Global Tic Severity Scale (YGTSS) was employed before therapy (baseline) and at 4, 8, and 12 weeks after therapy. RESULTS: (1) The total effect rates of treatment with a clonidine transdermal patch were 29.27, 53.66, and 63.41% at 4, 8, and 12 weeks, respectively. Compared with the baseline, the differences were significant at three different observation periods. (2) Compared to the level of 25% reduction, there were significant decreases in the score-reducing rate of motor tic and total tic severities at 12 weeks. (3) If the disease course was ≤24 months and the motor tic score was <16 at the baseline, there was an effective rate of 100% for treatment with the clonidine transdermal patch. If the disease course was ≤24 months and the motor tic score was >16, there was an effective rate of 57.1%. If the disease course was >24 months and the clinical classification was chronic TD, there was an effective rate of 62.5%. If the disease course was >24 months and the clinical classification was Tourette's syndrome, 90% of the patients were invalid. (4) The main adverse events were rash, slight dizziness, and headache. CONCLUSION: (1) When patients were pretreated with a D2-dopamine receptor antagonist that was ineffective or not tolerated well, switching to a clonidine transdermal patch treatment was effective and safe. (2) A clonidine transdermal patch could be a first-line medication for mild and moderate TD cases that are characterized by motor tics.

Dynamic Changes in Spectral and Spatial Signatures of High Frequency Oscillations in Rat Hippocampi during Epileptogenesis in Acute and Chronic Stages.

OBJECTIVE: To analyze spectral and spatial signatures of high frequency oscillations (HFOs), which include ripples and fast ripples (FRs, >200 Hz) by quantitatively assessing average and peak spectral power in a rat model of different stages of epileptogenesis. METHODS: The lithium-pilocarpine model of temporal lobe epilepsy was used. The acute phase of epilepsy was assessed by recording intracranial electroencephalography (EEG) activity for 1 day after status epilepticus (SE). The chronic phase of epilepsy, including spontaneous recurrent seizures (SRSs), was assessed by recording EEG activity for 28 days after SE. Average and peak spectral power of five frequency bands of EEG signals in CA1, CA3, and DG regions of the hippocampus were analyzed with wavelet and digital filter. RESULTS: FRs occurred in the hippocampus in the animal model. Significant dynamic changes in the spectral power of FRS were identified in CA1 and CA3. The average spectral power of ripples increased at 20 min before SE (p < 0.05), peaked at 10 min before diazepam injection. It decreased at 10 min after diazepam (p < 0.05) and returned to baseline after 1 h. The average spectral power of FRs increased at 30 min before SE (p < 0.05) and peaked at 10 min before diazepam. It decreased at 10 min after diazepam (p < 0.05) and returned to baseline at 2 h after injection. The dynamic changes were similar between average and peak spectral power of FRs. Average and peak spectral power of both ripples and FRs in the chronic phase showed a gradual downward trend compared with normal rats 14 days after SE. SIGNIFICANCE: The spectral power of HFOs may be utilized to distinguish between normal and pathologic HFOs. Ictal average and peak spectral power of FRs were two parameters for predicting acute epileptic seizures, which could be used as a new quantitative biomarker and early warning marker of seizure. Changes in interictal HFOs power in the hippocampus at the chronic stage may be not related to seizure occurrence.