Objective:To analyze antiviral activity against respiratory syncytial virus (RSV) of interferon (IFN)-α2b and IFN-λ1 on Hep2 cells and human airway epithelial (HAE) cells.Methods:IFN-α2b or IFN-λ1 was incubated with Hep2 cells after RSV infection, and 48 hours later, the cytopathic effect was observed, the viral load was determined using real time/reverse transcription quantitative polymerase chain reaction (RT qPCR), RSV F protein expression was detected using immunofluorescence, and cell survival rate was detected using crystal violet. HAE cells were incubated with IFN-α2b or IFN-λ1 for 24 hours, and then HAE were challenged with RSV. The viral load in the culture supernatant was determined on days 1-7 using RT qPCR, RSV F protein was determined with immunofluorescence and the viral titers in the culture supernatant was detected on day 7 by plaque assay.Results:In Hep2 cells, the CPE of the treatment groups (IFN-α2b and IFN-λ1) was alleviated compared to the virus control group, and the CPE of the high concentration group was lighter than that of the low concentration group. Different concentrations of IFN-α2b and IFN-λ1 could significantly reduce the viral load of RSV ( P<0.001), and the viral load of the high concentration group was significantly lower than that of the low concentration group ( P<0.001). In addition, IFN-α2b and IFN-λ1 could reduce the RSV F protein expression after RSV infection and improve cell survival rate. In HAE cells, IFN-α2b and IFN-λ1 could inhibit RSV virus replication, reduce virus titers ( P<0.001) and reduce RSV F protein expression. Conclusions:IFN-α2b and IFN-λ1 both showed great antiviral activity against RSV in Hep2 and HAE cells, providing data reference for the study of interferon against respiratory viruses.

Objective:To evaluate modified Lanza score (MLS) of gastric mucosa for predicting the prognosis of geriatric patients with sepsis.Methods:Data of 50 patients with sepsis, who were over 60 years old and underwent gastroscopy for suspected gastrointestinal bleeding in the Department of Geriatric Critical Care Medicine of Guangdong Provincial People's Hospital from January 2019 to April 2022, were retrospectively analyzed. Patients were divided into the death group ( n=32) and the survival group ( n=18) according to their regression within 28 days after gastroscopy. Their gastric mucosa was scored by using MLS system, and the mortality of patients with MLS≥1 was calculated, then the patients were further divided into 2 groups, MLS=0-2 ( n=23, less than 2 regions of lesions ) and MLS=3-5 ( n=27, two or more regions of lesions). The relationship between MLS and acute physiology and chronic health status evaluation (APACHE) Ⅱ score, risk factor of death and mortality in each group were compared. The correlation between MLS and mortality was analyzed. The influence of geriatric sepsis risk factors affecting the prognosis of patients within 28 days were analyzed by using logistic regression. Results:Among the 50 geriatric patients with sepsis, those with gastric mucosal lesions, i.e., MLS ≥1, accounted for 68.00% (34/50), including 84.38% (27/32) patients with MLS≥1 in the death group, which was significantly higher than the 38.89% (7/18) patients with MLS≥1 in the survival group ( χ 2=10.593, P<0.001). Patients with MLS=3-5 had significantly higher APACHE Ⅱ scores (26.09±6.47 VS 18.57±7.66, t=3.527, P=0.001) and higher mortality [85.19% (23/27) VS 39.13% (9/23), χ 2=11.434, P=0.001] compared with MLS=0-2. Correlation analysis showed a significant correlation between MLS and mortality ( r=0.886, P=0.019). Multivariate logistic regression analysis showed that MLS=4-5 was an independent risk factor for death in geriatric patients with sepsis ( OR=17.055, 95% CI: 1.387-209.744, P=0.027). Conclusion:MLS presents high sensitivity in predicting 28-day outcomes for geriatric patients with sepsis. Two or more than 2 regions of gastric mucosal lesions can significantly increase the risk of death in geriatric patients with sepsis.

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.

Objective:To investigate the epidemiological characteristics of human bocavirus 1 (HBoV1) and to analyze the genetic variation.Methods:A total of 2 848 nasopharyngeal aspirate (NPAs) specimens were collected from hospitalized children with acute respiratory tract infections (ARTI) in Beijing Friendship Hospital from April 2017 to March 2019, and HBoV1 was detected by quantitative real-time PCR. Epidemiological analysis was carried out based on the clinical information of the patients. The nested PCR method was used to amplify the NP1 and VP1 genes of HBoV1 for homology analysis. Maximum clade credibility tree (MCC tree) and genetic polymorphism map were constructed to analyze the time evolution of HBoV1 VP1.Results:HBoV1 was detected in 90(3.16%) of 2 848 NPAs, most (93.33%, 84/90) HBoV1-positive cases were among children <5 years of age. HBoV1 could be detected throughout the year with a higher prevalence 7.23% (18/249) in October. Of the 90 HBoV1-infected cases, the main clinical symptoms were fever and cough, 44(48.89%) were co-infected with other respiratory viruses; 55 NP1 sequences and 47 VP1 sequences were obtained by nested PCR amplification, phylogenetic analysis showed that the nucleotide homology was 98.9%~-100% and 99.1%~-100%, respectively. MCC tree showed that the HBoV1 VP1 gene sequence obtained in this study appeared in two adjacent clades, the gene evolution was stable.Conclusions:HBoV1 is one of the common viruses that cause respiratory infection among children in Beijing. HBoV1 genetic evolution is relatively stable, but it still needs to be monitored continuously.

Objective:To investigate the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of malignant endometrial lesions.Methods:A total of 142 patients with endometrial lesions who underwent contrast-enhanced ultrasound examination in Beijing Tiantan Hospital, Capital Medical University from January 2019 to September 2021 were selected. The endometrial lesions were divided into benign group (including endometrial hyperplasia and endometrial polyps) and malignant group (endometrial cancer) according to the pathological results of surgery, and the differences of contrast enhancement patterns between benign group and malignant group were compared. The sensitivity, specificity of CEUS in the diagnosis of endometrial cancer were calculated, and the Kappa value was calculated with the initial enhancement time earlier than or equal to the muscularity or the peak time earlier than the muscularity as the diagnostic index for the diagnosis of endometrial malignant lesions. The sensitivity, specificity, and Kappa value of CEUS in the diagnosis of endometrial cancer and endometrial cancer with thickness ≥10 mm were calculated.Results:A total of 108 patients underwent surgery with clear pathological results, including 66 patients in the benign lesion group and 42 patients in the malignant lesion group. The thickness of malignant lesions was significantly larger than that of benign lesions, and the difference was statistically significant( t=4.039, P<0.05), but there was no significant difference of hemodynamic parameters between the two groups ( P>0.05). The initial enhancement time, peak time and peak intensity of benign lesions were significantly different from those of malignant lesions(all P<0.05). The sensitivity, specificity, and Kappa value of CEUS in the diagnosis of endometrial cancer were 64.3%, 100% and 0.668, respectively. The sensitivity, specificity and Kappa value of CEUS in the diagnosis of endometrial carcinoma with lesion thickness ≥10 mm were 75.0%, 100% and 0.795, respectively. Conclusions:For the diagnosis of endometrial lesions, especially the malignant endometrial lesions with thickness and diameter greater than or equal to 10mm, there is a high diagnostic coincidence rate between CEUS and pathological diagnosis, and endometrial malignant lesions have more specific CEUS manifestations.

Objective:To find clues potentially valuable for fighting against infection with human respiratory syncytial virus (HRSV), the differentially expressed genes in HEp-2 cells infected with HRSV were analyzed.Methods:Gene expression profiles of HEp-2 cells infected with HRSV were collected from the public gene expression omnibus (GEO) database. The differentially expressed genes following HRSV infection at each time point of 4, 8, 12, and 15 hours were found using R language. The differentially expressed genes were analyzed by gene ontology (GO), KEGG pathway and protein-protein interaction network (PPI). Genes with relatively high protein interaction in PPI were randomly selected for quantitative reverse transcription-polymerase chain reaction (qRT-PCR) verification at the transcription level from HEp-2 cells after HRSV infection at 4 hours.Results:A total of 101 differentially expressed genes were determined, including 92 upregulated genes and 9 downregulated genes. Function enrichment analysis revealed that HRSV infection could cause significant changes in multiple signaling pathways such as immune response in HEp-2 cells. The results of qRT-PCR were consistent with the trend of transcriptome data.Conclusions:The differentially expressed genes and the change of signaling pathways in HRSV-infected HEp-2 cells is of great significance to the studies on pathogenic mechanism and prevention of HRSV infection.

Objective:To investigate the characteristics of three-dimensional high resolution anorectal manometry and the effect of biological feedback therapy on the improvement of clinical symptoms in very elderly patients with chronic functional constipation.Methods:A total of 68 cases with chronic functional constipation were divided into very elderly group(≥80 years old, n=36)and the elderly group(60-79 years old, n=32)in the retrospective analysis.Patients underwent the three-dimensional high resolution anorectal manometry before and after biological feedback therapy, and the related parameters and scores of constipation symptoms before and after treatment were compared between the two groups.Results:The results of 3D high-resolution anorectal manometry showed that the anus relaxation rate in the simulated defecation test was lower and the bowel threshold in the rectal sensory threshold test was higher in the very elderly group than in the elderly group(2.44±33.81% vs.16.34±16.99%, 103.44±42.01 ml vs.77.22±41.85 ml, t=-2.047 and 2.655, P=0.049 and 0.012). In the very elderly group, the post-biological feedback therapy versus pre-biological feedback therapy showed that anal residual pressure during simulated defecation was decreased, the absolute value of negative anorectal pressure difference was reduced and the anal relaxation rate was increased [57.50±18.88 mmHg(1 mmHg=0.133 kPa) vs.64.84±25.82 mmHg, -29.64±15.98 mmHg vs.-39.47±19.45 mmHg, 10.53±29.35% vs.2.44±33.81%, t=3.342, -4.902 and -3.209, P=0.002, 0.000 and 0.003]. The scores of clinical symptom scale showed that there was no significant difference in the effective rate between the very elderly and elderly groups(66.67% or 24/36 vs.71.88% or 23/32, χ2=0.760, P=0.860). Conclusions:The elderly functional constipation patients with defecation disorder often have rectal propulsive insufficiency and dyscoordination of pelvic floor muscle contraction.The main cause of defecation disorder in very elderly patients is the decrease of anal relaxation rate during simulated defecation.Biological feedback therapy can improve the symptoms of defecation disorder in very elderly patients by reducing the anal residual pressure during simulated defecation, increasing the anal relaxation rate and reducing the absolute value of negative anorectal pressure difference.

Objective:To evaluate the effectiveness of three laboratory routine disinfection methods on influenza virus inactivation.Methods:The physical method of ultraviolet irradiation and two chemical methods of ethanol and "84 disinfectant" were used to inactivate the influenza virus respectively. After disinfecting the virus samples at different times or concentrations, MDCK cells were inoculated and the inactivation effect of influenza virus was identified by observing cytopathic effect (CPE).Results:When the influenza virus was irradiated by ultraviolet light for ≥1 min, treated with 75% ethanol for 0.25-32 min, the effective chlorine concentration of "84 disinfectant" was ≥400 mg/L, and the influenza virus was disinfected at room temperature for ≥1 min, no virus infectivity could be detected under the three conditions.Conclusions:The condition of inactivating influenza virus by routine disinfection methods in laboratory was explored, which provided reference data for establishing the protection specifications and the emergency treatment plans for influenza virus laboratories.

Objective:To study the sequence characteristics and genetic variation of a human respiratory syncytial virus (HRSV) subtype A genome in Beijing.Methods:The genomic RNA of HRSV from nasopharyngeal aspirate samples was sequenced and obtained a whole genome sequence of HRSV A subtype. The phylogenetic tree was constructed with reference sequences of other HRSV strains. The major proteins were compared and single nucleotide polymorphism analyzed. In addition, the N-glycosylationsites of F and G protein were predicted.Results:Phylogenetic tree and homology analysis results suggest that the HRSV strain (RSVA/Beijing-China/2017) was the A subtype ON1 genotype. Nucleotide and amino acid variation analysis showed that G protein, F protein and L protein had some substitutions. Analysis of amino acid variation sites showed that amino acid substitution (L142S) occurred at position 142 of G protein. For F protein, there were two substitutions, which were S105N in the P27 peptide (110-136aa) and C69Y in the antigen sites ? (62-69 aa and 196-210 aa). The prediction of N-glycosylation sites revealed that there were 5 N-glycosylation sites of F protein and 4 N-glycosylation sites of G protein in this strain.Conclusions:The HRSV strain obtained in Beijing belongs to A subtype ON1 genotype. The G, F and L proteins have large variations, and 22 amino acid substitutions have occurred in the G and F proteins.

Objective: To evaluate the efficacy and safety of Oryz-Aspergillus enzyme and pancreatin tablets (Combizym^®) in the treatment of postprandial distress syndrome (PDS) in the elderly, compared with gastrointestinal motility drugs. Methods: A prospective randomized controlled trial was designed and registered in the China Clinical Trials Registry (ChiCTR-IPR-16008185). The elderly patients with PDS were randomly divided into three groups, including Mosapride group with Mosapride citrate tablets 5 mg 3 times per day for 2 weeks; Combizym^® group with Combizym tablets 244 mg 3 times per day for 2 weeks; combined treatment group with both drugs and same doses for 2 weeks. The modified Nepean dyspepsia index (NDSI) score, discomfort intensity score and PDS score were calculated on patients before treatment, at the end of first and second week of treatment, as well as 4 weeks after treatment finished, respectively. Adverse effects were evaluated. Results: A total of 323 patients from 16 tertiary hospitals in China were enrolled in this study. Among them, 105 patients were in Mosapride group, 109 in Combizym^® group and 109 in combined treatment group. There were 148 males (45.8%) and 175 females (54.2%) with median age 71.4±9.0 years (60-100 years). Baseline characteristics of three groups were comparable. After treatment, the NDSI scores in three groups all decreased significantly (P<0.001), while they were similar between groups (P>0.05). The discomfort intensity score and PDS score in three groups showed a significant reduction after treatment (P<0.001), especially in the combined treatment group. Compared with Mosapride group, the scores in Combizym^® group decreased significantly after one or two weeks [discomfort intensity score: after one week, 4.0(2.5, 8.0) vs. 6.0(3.0, 10.0); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 6.0); all P<0.05. PDS score: after one week, 6.0(3.0, 9.0) vs. 7.0(3.5, 10.5); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 7.0); all P<0.05]. The efficacy rate in all patients after first week of treatment was over 15.0%. The efficacy rates after two weeks were 55.2%, 68.8% and 73.4% in Mosapride group, Combizym^® group and combined treatment group, respectively. After two week treatment, the efficacy rates in Combizym^® group (P=0.041) and combined group (P=0.006) were higher than that of Mosapride group. The recurrence rate of Mosapride group was 9.5%, which was significantly higher than that of Combizym^® group (1.8%, P<0.05) and combined treatment group (1.8%, P<0.05). There were no serious adverse effects in the three groups. Conclusions The efficacy of Oryz-Aspergillus enzyme and pancreatin tablets is comparable with that of Mosapride in elderly PDS patients, with fewer adverse effects and low recurrence rate. Combination regimen indicates better efficacy than that of Oryz-Aspergillus enzyme and pancreatin tablets or Mosapride alone.