The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.
Base Sequence ; Blotting, Western ; Coronavirus NL63, Human ; chemistry ; Fluorescent Antibody Technique, Indirect ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; Spike Glycoprotein, Coronavirus ; Vaccinia virus ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

Objective:To compare the clinical characteristics of patients with non-erosive gastroesophageal reflux disease (NERD), reflux hypersensitivity (RH) and functional heartburn (FH), in order to help the differential diagnosis and treatment of the three diseases.Methods:From October 2016 to February 2019, 64 patients with persistent heartburn who had negative endoscopy in the Department of Gastroenterology, the Second Affiliated Hospital of Xi′an Jiaotong University and with negative results of endoscopy examination were consecutively selected, which included 26 NERD patients (NERD group), 12 RH patients (RH group) and 26 FH patients (FH group). The gender, age, body mass index (BMI), presence of hiatus hernia, proton pump inhibitor (PPI) treatment effect, the score and incidence of gastroesophageal symptoms within one month before visit, and the incidence of overlapping functional dyspepsia (FD), postprandial distress syndrome (PDS) and epigastric pain syndrome (EPS) of the three groups were compared. One-way analysis of variance, least-significant difference, Kruskal-Wallis H test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:There were no statistically significant differences in gender, age or incidence of esophageal hiatal hernia between NERD group, RH group and FH group (all P>0.05). BMI of NERD group was higher than that of FH group ((23.74±3.10) kg/m 2 vs. (21.26±2.75) kg/m 2), and the difference was statistically significant ( t=3.066, P=0.003). The effective rate of PPI treatment in NERD group was higher than those of RH group and FH group (19, 4, and 11 cases, respectively), the differences were statistically significant ( χ2=5.428 and 5.042, P=0.020 and 0.025). The score and incidence of burning sensation in upper abdomen of NERD group were both lower than those of FH group (0, 0 to 0 vs. 0, 0 to 5; 7.7%, 2/26 vs. 38.5%, 10/26), and the differences were statistically significant ( Z=-2.756, P=0.006; χ2=6.933, P=0.008). There were no statistically significant differences in the score of heartburn symptoms, or the scores and incidence of chest pain, reflux, globular sensation of hysteria, burning sensation of throat, cough, epigastric pain, early satiety or post meal fullness, belching, nausea or vomiting between NERD group, RH group and FH group (all P>0.05). There were no statistically significant differences in the incidence of overlapping FD or overlapping PDS between the NERD group, RH group and FH group (both P>0.05), but the incidence of overlapping EPS of FH group was higher than that of NERD group (46.2%, 12/26 vs. 11.5%, 3/26), and the difference was statistically significant ( χ2=7.589, P=0.013). Conclusions:There are certain differences in the clinical characteristics of patients with NERD, RH or FH, mainly reffected in the PPI treatment effects and whether overlapping EPS. The higher incidence of FH overlapping EPS further strengthens the hypothesis that functional gastrointestinal disorders have a common pathophysiological mechanism.

In this paper, some significant problems, which were found frequently in the products of autoimmune
Indicators and Reagents

Reference interval study and clinical evaluation are crucial supportive researches to demonstrate the intended use of quantitative
Indicators and Reagents ; Reference Values

OBJECTIVE: To analyze the differences in reflux patterns in 24-hour esophageal pH-impedance monitoring in patients with non-erosive reflux disease (NERD), reflux hypersensitivity (RH) and functional heartburn (FH) and explore the possible mechanism of symptoms in patients with heartburn and negative endoscopic findings. METHODS: Seventy-nine patients with heartburn as the main symptoms but negative endoscopic findings, including 35 with NERD, 16 with RH and 28 with FH, were enrolled in this study.All the patients underwent 24-h esophageal pH-impedance monitoring and esophagogastroscopy, and the results were compared among the 3 groups. RESULTS: Acid reflux episode was significantly increased and weakly alkaline reflux episode was significantly decreased in NERD group in comparison with RH group and FH group ( CONCLUSIONS Patients with NERD, RH and FH had different reflux patterns.Acid reflux is predominant in the NERD, while weakly alkaline reflux is significantly increased RH and FH.In patients with normal esophageal acid exposure but without symptoms or without recorded symptoms during esophageal pH-impedance monitoring, analysis of the total reflux episode, mixed reflux episode, proximal acid reflux episode and percentage can help in the differential diagnosis between RH and FH.
Electric Impedance ; Esophageal pH Monitoring ; Gastroesophageal Reflux/diagnosis* ; Heartburn/etiology* ; Humans ; Hydrogen-Ion Concentration

OBJECTIVE: To investigate the value of positive lymph node ratio (LNR) in predicting the prognosis of patients with esophageal cancer. METHODS: We retrieved the data of a total of 862 patients with esophageal cancer with complete clinical pathology data archived in SEER database in 2010 to 2015. The best cutoff point of LNR was selected using X-tile software. Univariate and multivariate COX proportional hazard models were used to assess the value of LNR in predicting the prognosis of patients after propensity score matching (PSM). RESULTS: The best cut-off point of LNR determined using X-tile 3.6.1 software was 0.16. The patients with LNR < 0.16 and those with LNR≥0.16 showed significant differences in the number of positive lymph nodes, pathological type, T stage and M stage. After 1:1 propensity score matching, the two groups showed no significant difference in the clinical data or pathological parameters. Matched univariate and multivariate COX regression analyses showed that LNR, primary tumor site and M staging were all independent risk factors affecting the prognosis of patients, and among them LNR had the most significant predictive value (LNR < 0.16 LNR≥0.16: HR=1.827, 95% : 1.140-2.929; =0.000). The median survival time of patients with LNR < 0.16 was 31 months (95%: 22.556-39.444 months), as compared with 16 months (95%: 12.989-19.011) in patient with LNR≥0.16 (Log Rank χ=27.392, < 0.0001). LNR had a better accuracy than N stage for assessing the patients' prognosis with an area under the ROC curve of 0.617 (95%: 0.567-0.666), as compared with 0.515 (95%: 0.463-0.565) of N stage (=3.008, =0.0026). CONCLUSIONS LNR≥0.16 is an independent risk factor affecting the prognosis of patients with esophageal cancer and has better prognostic value than N stage.
Esophageal Neoplasms ; Humans ; Lymph Node Excision ; Lymph Node Ratio ; Lymph Nodes ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Risk Factors