Objective To investigate the clinical prognosis of idiopathic demyelinating optic neuritis (IDON), the rate of its conversion to multiple sclerosis (MS) or neuromyelitis optica (NMO) and its clinical features related to the conversion.Methods Patients satisfying our entry criteria for IDON hospitalized in Beijing Tongren Hospital during the period from 2002 to 2007 were re-evaluated with follow-ups for 6-months to 5-years.The McDonald diagnostic criteria for MS (revised, 2005) was used to diagnose MS in these subjects during follow-up and the diagnosis of NMO utilized 1999 Wingerchuk' s NMO criteria.The Chi-Squared χ2 test was applied to statistically analyze association of clinical features and development of MS or NMO.Results In 107 recruited IDON cases with complete clinical data and follow-up, 12 cases (11.2%) developed into MS or NMO during follow-up period.All 12 cases met the revised McDonald criteria, of which 4 cases met NMO criteria and the remaining eight cases showed some clinical evidence of "optic-spinal MS (OS-MS)".A significantly higher conversion rate of 23.1% was found in recurrent IDON than the 4.4% in single-episode cases (χ2 = 6.899, P < 0.01) .Convesion rate of female patiends (17.2%) is significantly higher than male patients (4.1%, χ2 = 4.620, P < 0.05).Conversion rate of 18.2% in patients with initially abnormal brain MRI was higher than rate of 8.1% in cases with normal brain MRI, but the difference was not statistically significant.No difference was found between presence or absence of swollen disc, nor severity of vision loss.Conclusions In a group of IDON patients, 11.2% developed into MS and NMO or clinically indicative OS-MS.Recurrent IDON and female gender suggested higher risk of developing MS or NMO.

Objective To construct c-Met CAR-T cells secreting PD-1 antibodies to reduce immune inhibitory effect of tumor cells and enhance the efficacy of CAR-T cell therapy against pancreatic cancer.Methods Kaplan-Meier Plotter,GEPIA,and Timer 2.0 bioinformatics databases were used to analyze c-Met expression in pancreatic cancer and its correlation with survival and immune infiltration status.In clinical samples of pancreatic cancer and pancreatic cancer Aspc-1 cells,c-Met and PD-L1 expressions were detected using immunohistochemistry or flow cytometry.Using gene editing technology,PD-1 secretory antibodies and HIS tags were linked to second-generation c-Met CAR molecules to construct PD-1/c-Met CAR plasmids,which were then packaged into lentiviruses for infection of activated T cells.The positive rate and cell subset distribution of CAR-T cells were analyzed with flow cytometry,and secretory PD-1 antibodies in cell supernatants were detected using Western blotting.The target cell killing efficiency and proliferative activity of the modified CAR-T cells were evaluated after activation,and cytokine secretion was analyzed using ELISA.Results The expression of c-Met was significantly higher in pancreatic cancer than in normal tissues,and its expression level was negatively correlated with the patients'survival and positively correlated with immune cell infiltration.The clinical samples of pancreatic cancer tissues expressed significantly higher levels of c-Met and PD-L1 than the adjacent tissues,and 90.7%and 57.7%of Aspc-1 cells were positive for c-Met and PD-L1,respectively.The constructed PD-1/c-Met CAR-T cells were capable of secreting PD-1 antibodies and showed a significantly higher killing efficiency against tumor cells than c-Met CAR-T cells at an effector-to-target ratio of 20:1,with also a higher proliferative activity after target cell stimulation and higher levels of IL-2 and TNF-α secretin.Conclusion PD-1/c-Met CAR-T cells have higher killing efficiency against pancreatic cancer cells with also higher proliferative activity than c-Met CAR-T cells.

Objective To construct c-Met CAR-T cells secreting PD-1 antibodies to reduce immune inhibitory effect of tumor cells and enhance the efficacy of CAR-T cell therapy against pancreatic cancer.Methods Kaplan-Meier Plotter,GEPIA,and Timer 2.0 bioinformatics databases were used to analyze c-Met expression in pancreatic cancer and its correlation with survival and immune infiltration status.In clinical samples of pancreatic cancer and pancreatic cancer Aspc-1 cells,c-Met and PD-L1 expressions were detected using immunohistochemistry or flow cytometry.Using gene editing technology,PD-1 secretory antibodies and HIS tags were linked to second-generation c-Met CAR molecules to construct PD-1/c-Met CAR plasmids,which were then packaged into lentiviruses for infection of activated T cells.The positive rate and cell subset distribution of CAR-T cells were analyzed with flow cytometry,and secretory PD-1 antibodies in cell supernatants were detected using Western blotting.The target cell killing efficiency and proliferative activity of the modified CAR-T cells were evaluated after activation,and cytokine secretion was analyzed using ELISA.Results The expression of c-Met was significantly higher in pancreatic cancer than in normal tissues,and its expression level was negatively correlated with the patients'survival and positively correlated with immune cell infiltration.The clinical samples of pancreatic cancer tissues expressed significantly higher levels of c-Met and PD-L1 than the adjacent tissues,and 90.7%and 57.7%of Aspc-1 cells were positive for c-Met and PD-L1,respectively.The constructed PD-1/c-Met CAR-T cells were capable of secreting PD-1 antibodies and showed a significantly higher killing efficiency against tumor cells than c-Met CAR-T cells at an effector-to-target ratio of 20:1,with also a higher proliferative activity after target cell stimulation and higher levels of IL-2 and TNF-α secretin.Conclusion PD-1/c-Met CAR-T cells have higher killing efficiency against pancreatic cancer cells with also higher proliferative activity than c-Met CAR-T cells.

Objective:To investigate the roles of N6-methyladenosine (m 6A) modification in regulating RP11-426A6.5 in the development of laryngeal squamous cell carcinoma (LSCC). Methods:The methylation and expression levels of lncRNAs were identified and important lncRNAs were screened utilizing long non-coding RNA (lncRNA) m 6A methylation microarray. Cancer and para cancer tissue samples were taken from 48 LSCC patients hospitalized to the Department of Otolaryngology-Head and Neck Surgery of the Second Affiliated Hospital of Harbin Medical University between January and September 2017. Expression profiling microarray was performed in 3 of 48 LSCC samples, and methylated RNA immunoprecipitation-quantitative PCR (MeRIP-qPCR) and quantitative real-time fluorescent PCR (qRT-PCR) were performed in the remaining 45 LSCC samples to verify the m 6A modification and expression levels of RP11-426A6.5. Correlations between RP11-426A6.5 and clinical factors were anlysed. Laryngeal cancer cell line with low expression of RP11-426A6.5 was created in vitro using RNA interference (RNAi) technology. The 5-Ethynyl-2′-deoxyuridine (EdU) cell proliferation experiment, wound healing experiment, and transwell invasion experiment were used respectively to measure the proliferation, migration, and invasion of LSCC cells. The effect of RP11-426A6.5 down-regulation on the growth of transplanted tumors in vivo was verified by nude mice tumorigenesis assay. The Cancer Genome Atlas (TCGA) database and sequence-based RNA adenosine methylation site predictor (SRAMP) website were used to predict the enzymes and corresponding methylation sites. MazF digestion was chosen to validate the binding sites. RNAi technology was used to observe the changes in cell function after interfering with the expression of the corresponding genes of the modified enzymes. MeRIP-qPCR was used to detect the level of RP11-426A6.5 m 6A cell line treated with actinomycin D was used to observe the stability of RP11-426A6.5. Results:RP11-426A6.5 methylation and expression levels were significantly higher in LSCC tissues than those in paracancerous tissues (methylation levels: 23.828±4.975 vs 20.280±3.607; expression levels: 1.197±0.314 vs 1.015±0.170, all P values<0.05). RP11-426A6.5 expression levels were closely correlated with T stage (T1-2: 1.081±0.298 vs T3-4: 1.306±0.292, χ 2=5.35, P<0.05). The postoperative survival of patients with high RP11-426A6.5 expressions was significantly lower than that of patients with low RP11-426A6.5 expression ( P=0.046). Assays in vitro and in vivo showed that the downregulation of RP11-426A6.5 significantly decreased the proliferation, migration, and invasion abilities of LSCC cells and the growth of transplanted tumors. The binding of methyltransferase-like 3 (METTL3), an m 6A-modified enzyme, to the corresponding methylation site of RP11-426A6.5 enhanced its stability and mediated its regulation of malignant behaviors of LSCC cells. Conclusions:RP11-426A6.5 can regulate the malignant behaviors of LSCC cells, which is mediated by the m 6A modification process involving in the methyltransferase METTL3.

Objective To investigate the neural mechanisms of speech motor control in individuals with Parkinson's disease (PD) under different strategies of motor execution control.MethodsThe techniques of the psychoacoustic and event-related potentials (ERPs) based on the altered auditory feedback paradigm were used in the present study. Two groups, including 17 PD patients and 17 healthy controls, were instructed to produce sustained vowels under the internal and external cueing tasks while hearing their voice randomly pitch-shifted downwards. The vocal responses and associated ERPs were recorded and compared across the groups and tasks.ResultsBehavioral results showed that the amplitude of acoustic compensation response was larger in PD patients than in the healthy controls (F=5.415, P=0.027), however, the main effect was not significant in the tasks (F=0.039, P=0.840). At the cortical level, PD patients produced significantly larger N1 responses to pitch perturbations in the internal cueing task related to the external cueing task (F=8.634, P=0.006), while such task effect was not observed in the healthy controls (F=1.550, P=0.231). Also, PD patients produced significantly larger N1 responses than the healthy controls in the internal cueing condition (F=5.476, P=0.026), but not in the external cueing condition (F=0.249, P=0.621). Conclusion Speech motor control in PD can be influenced by the strategies of motor execution control. Compared to the internal cueing task, the external cueing task can increase neural efficiency in the encoding of speech auditory feedback PD patients that improve auditory-motor integration for speech production.