Objective Our previous studies established that microRNA (miR)-451 from human umbilical cord mesenchymal stem cell-derived exosomes (hUC-MSC-Exos) alleviates acute lung injury (ALI). This study aims to elucidate the mechanisms by which miR-451 in hUC-MSC-Exos reduces ALI by modulating macrophage autophagy. Methods Exosomes were isolated from hUC-MSCs. Severe burn-induced ALI rat models were treated with hUC-MSC-Exos carrying the miR-451 inhibitor. Hematoxylin-eosin staining evaluated inflammatory injury. Enzyme-linked immunosorbnent assay measured lipopolysaccharide (LPS),tumor necrosis factor-α,and interleukin-1β levels. qRT-PCR detected miR-451 and tuberous sclerosis complex 1 (TSC1) expressions. The regulatory role of miR-451 on TSC1 was determined using a dual-luciferase reporter system. Western blotting determined TSC1 and proteins related to the mammalian target of rapamycin (mTOR) pathway and autophagy. Immunofluorescence analysis was conducted to examine exosomes phagocytosis in alveolar macrophages and autophagy level. Results hUC-MSC-Exos with miR-451 inhibitor reduced burn-induced ALI and promoted macrophage autophagy. MiR-451 could be transferred from hUC-MSCs to alveolar macrophages via exosomes and directly targeted TSC1. Inhibiting miR-451 in hUC-MSC-Exos elevated TSC1 expression and inactivated the mTOR pathway in alveolar macrophages. Silencing TSC1 activated mTOR signaling and inhibited autophagy,while TSC1 knockdown reversed the autophagy from the miR-451 inhibitor-induced. Conclusion miR-451 from hUC-MSC exosomes improves ALI by suppressing alveolar macrophage autophagy through modulation of the TSC1/mTOR pathway,providing a potential therapeutic strategy for ALI.

Objective:To understand the composition of respiratory virus spectrum in hospitalized children with acute respiratory infections in Guilin City from 2021 to 2022, and to explore the epidemic characteristics of different respiratory viruses.Methods:Information of a total of 638 hospitalized children with acute respiratory infections (ARI) syndrome under the age of 14 years in Guilin city, Guangxi from September 2021 to October 2022 was collected as research data. The collected pharyngeal swabs from pediatric patients were subjected to real-time fluorescence reverse transcription polymerase chain reaction for nucleic acid testing, screening for 15 common respiratory viruses, and analyzing the virus spectrum characteristics of hospitalized pediatric patients with acute respiratory infections.Results:Among the 638 specimens, 365 were tested positive for at least one virus, with a positive rate of 57.21% (365/638). The detection rate for two or more viruses was 12.85% (82/638). There were 12 types of viruses detected, and the difference in virus detection rates among different seasons was statistically significant (P<0.002). The virus positive detection rate of samples from different age groups was the highest in the 0-2-year old group (40.66%), followed by the 3-5-year old group (34.80%) and the 6-year and older group (24.54%).Conclusions:During the period from September 2021 to October 2022, all 12 respiratory viruses were prevalent in Guilin area, with obvious summer peak as characteristics. It is suggested that corresponding prevention and control measures should be focused on pathogens that may cause acute respiratory infections in children during the local summer to prevent and control the spread and prevalence of respiratory infections.

Objective:To understand the prevalence and characteristics of Rhinovirus (HRV) infection in influenza-like Illness (ILIs) patients in Mianyang, Sichuan province, China.Methods:Throat swabs were collected from patients of ILIs in sentinel hospitals in Mianyang during 2021—2022. Real-time fluorescence quantitative PCR was used to detect 16 common pathogens. The VP4/VP2 coding region genes of HRV positive samples were amplified by nest PCR. The phylogeny, consistency and amino acid variation of different serotypes were analyzed and compared with reference sequences from GenBank database.Results:A total of 332 ILIs′ samples were collected with a virus detection rate of 58.73% (195/332) in Mianyang. Among them, 23 samples (23/332) were HRV-positive, and 18 VP4/VP2 sequences of HRV strains were successfully amplified. It was found that 13 HRV serotypes were detected in ILIs samples in Mianyang, which belonged to three genotypes, namely HRV-A (12 strains), HRV-B (5 strains) and HRV-C (1 strain).Conclusions:HRV was one of the pathogens of ILIs cases in Mianyang during 2021—2022, with HRV-A types as the dominant strains.

Objective:To establish a method via transfection of RNA to rescue coxsackievirus B3 B3 (CVB3).Methods:The efficiency of CVB3 genomic RNA extraction from three nucleic acid extraction reagents, Qiagen 57704, Qiagen 52904, and Trizol, and the transfection efficiency of viral RNA with two transfection reagents (Lipofectamine 2000 and Lipofectamine 3000) were compared. The efficiency of CVB3 rescue in Vero cells and HEK293T cells to determine the optimal conditions for rescuing CVB3.Results:The number of phagolysosomes for virus rescue by Qiagen 57704, Qiagen 52904, and Tizol kit extracted RNA was 13.33±1.53, 150±15.00, and 1.67±0.58, respectively, and there was a statistically significant difference in the efficiency of the three method of extracting CVB3 RNA to rescue the viral RNA ( F=268.920, P<0.001); The number of phage spots formed by Lipofectamine3000 and Lipofectamine2000 transfected RNA was 74.50±3.00 and 22.00±5.00, respectively, and the difference was statistically significant ( P<0.01); Qiagen 52904 reagent extracted CVB3 nucleic acid more efficiently than Qiagen 57704 and Trizol reagents; the transfection efficiency of transfection reagent Lipofectamine 3000 was 3 times more than than that of Lipofectamine 2000, and the efficiency of virus rescue of CVB3 in HEK293T cell culture was higher than that of HeLa and Vero cells, and the copy numbers of the three kinds of viruses rescuing the virus were 6.09×10 7±8.00×10 5, 5.18×10 3±6.17×10 2 and 0, the difference was statistically significant ( F=17 383.644, P<0.001), and it was also found that the efficiency of virus rescue could be improved by multiple elution when extracting RNA. Conclusions:In this study, we successfully established the method of transfecting RNA to rescue CVB3, which can effectively improve the efficiency of virus rescue by choosing Qiagen 52904 nucleic acid extraction kit, increasing the number of elution, using Lipofectamine 3000 transfection reagent, and transfection of HEK293T cells.

Objective:To investigate the etiology of hand, foot and mouth disease (HFMD) in Mianyang city in 2022, and to analyze the genetic characteristics of coxsackievirus A6.Methods:Pharyngeal swabs were collected from patients with HFMD in Mianyang city in 2022. Real-time fluorescence quantitative PCR was used to detect enteroviruses in the samples. Part of the VP1 gene in enterovirus-positive samples was amplified by nested PCR using enterovirus typing primers to further identify the viral types. The VP1 coding region of all CVA6-positive samples and the whole genome of some samples were amplified and sequenced by PCR. The endemic strain in Mianyang city was analyzed for phylogeny, gene homology, amino acid variation and genetic recombination.Results:A total of 151 pharyngeal swabs were collected, and 104 enterovirus-positive samples were detected by real-time fluorescence quantitative PCR, with an overall detection rate of 68.88% (104/151). The typing results showed that there were 77 cases of CVA6 infection, with a positive rate of 50.99% (77/151). The full-length VP1 genes of 77 CVA6 strains were amplified, sequenced, and successfully spliced, and phylogenetic analysis showed that all 77 strains were of the D3 genotype. There were multiple amino acid variant sites in the prevalent strains in Mianyang city compared with the reference strain. Twenty whole genome sequences were amplified, sequenced, and successfully spliced, and homology analysis showed that the nucleotide homology between the 20 positive sequences ranged from 97.0% to 99.9%. Phylogenetic tree and recombination analysis showed that no recombination occurred in the coding regions of the epidemic strains in this study.Conclusions:The predominant pathogen causing HFMD in Mianyang city in 2022 is CVA6 D3 subtype, which is consistent with the national epidemic in 2022.

BACKGROUND: Immune checkpoint inhibitors (ICIs) are increasingly used as first-line therapy for patients with advanced non-small cell lung cancer (NSCLC) harboring no actionable mutations; however, data on their efficacy among patients presenting with intracranial lesions are limited. This study aimed to explore the efficacy and safety of ICIs combined with chemotherapy in advanced NSCLC patients with measurable brain metastasis at initial diagnosis. METHODS: Our study retrospectively analyzed clinical data of a total of 211 patients diagnosed with driver gene mutation-negative advanced NSCLC with measurable, asymptomatic brain metastasis at baseline from Hunan Cancer Hospital between January 1, 2019 and September 30, 2021. The patients were stratified into two groups according to the first-line treatment regimen received: ICI combined with chemotherapy ( n = 102) or chemotherapy ( n = 109). Systemic and intracranial objective response rates (ORRs) and progression-free survival (PFS) were analyzed. Adverse events were also compared between the groups. RESULTS: Compared with the chemotherapy-based regimen, the ICI-containing regimen was associated with a significantly higher intracranial (44.1% [45/102] vs . 28.4% [31/109], χ2 = 5.620, P = 0.013) and systemic (49.0% [50/102] vs . 33.9% [37/109], χ2 = 4.942, P = 0.019) ORRs and longer intracranial (11.0 months vs . 7.0 months, P <0.001) and systemic (9.0 months vs . 5.0 months, P <0.001) PFS. Multivariable analysis consistently revealed an independent association between receiving ICI plus platinum-based chemotherapy as a first-line regimen and prolonged intracranial PFS (hazard ratio [HR] = 0.52, 95% confidence interval [CI]: 0.37-0.73, P <0.001) and systemic PFS (HR = 0.48, 95% CI: 0.35-0.66, P <0.001). No unexpected serious adverse effects were observed. CONCLUSION: Our study provides real-world clinical evidence that ICI combined with chemotherapy is a promising first-line treatment option for driver gene mutation-negative advanced NSCLC patients who present with brain metastasis at initial diagnosis. CLINICAL TRIAL REGISTRATION https://www.clinicaltrials.gov/ , OMESIA, NCT05129202.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Immune Checkpoint Inhibitors/therapeutic use* ; Retrospective Studies ; Brain Neoplasms/genetics*

Objective The expression and prognosis significance of CD59 in pancreatic cancer were analyzed by bioinformatics.Methods Gene expression profiling interactive analysis 2(GEPIA2)and human protein atlas(HPA)databases were used to compare the expression of CD59 between pancreatic cancer tissues and adjacent tissues;Kaplan-Meier plotter database was used to evaluate the effect of CD59 on prognosis;String and Cytoscape3.9.1 were used to analyze the CD59 protein interaction network;DAVID6.8 performed gene enrichment and pathway enrichment analysis of CD59 with key interacting genes.Results Compared with normal tissues,the expression of CD59 in pancreatic cancer tissues was significantly up-regulated(P<0.05),and the overall survival time(HR=2.3,95%CI:1.52-3.50)and recurrence free survival(HR=4.31,95%CI:1.57-11.83)of pancreatic cancer patients with CD59 high expression was shorter than CD59 low expression patients.Protein interaction network analysis revealed that CD59 is closely related to several molecules such as CD55,GOLGA2,LMAN1,TMED2 and SERPINA1.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis indicated that CD59 was mainly involved in pathways such as complement activation,innate immune response and coronavirus-COVID-19.Conclusion CD59 is highly expressed in pancreatic cancer tissue and is affected by several immune-related genes.It is associated with poor prognosis of patients and can be used as one of biomarkers for early diagnosis and prognosis prediction of pancreatic cancer.

Objective:Continuous glucose monitoring (CGM) technology is used to compare the advantages of insulin degludec (IDeg) as a basal insulin regimen compared with insulin glargine (IGlar) in the treatment of adult type 1 diabetes mellitus.Methods:30 adult patients with T1DM admitted to Heji Hospital Affiliated to Changzhi Medical College from September 2019 to December 2020 were screened. According to the random number table method, the patients were randomly divided into two groups (insulin degludec group and insulin glargine group) at a ratio of 1∶1, respectively treated with IDeg, IGlar and aspartate insulin for 12 weeks. The main outcome measures were the coefficient of variation of blood glucose (CV), mean amplitude of glycemic excursions (MAGE), time in range (TIR), time above range (TAR) and time below range (TBR). The secondary outcome measures were mean blood glucose (MBG), standard deviation of blood glucose (SD), fasting blood glucose (FPG), 2 h postprandial blood glucose (2 h BG), hemoglobin A1c (HbA 1c), means of daily differences (MOOD), and the frequency of hypoglycemic events. Results:At 12 weeks of treatment, the HbA 1c, FPG, 2 h BG, MBG, SD, CV and MAGE of insulin degludec group were lower than those of insulin glargine group, with statistically significant difference (all P<0.05). The TIR in the insulin degludec group was significantly higher than that in the insulin glargine group [73(63, 75)% vs 43(28, 63)%, P<0.001], and the TAR was lower than that in the glycerine group [25(17, 23)% vs 35(33, 64)%, P=0.003]. From the curve spectrum of blood glucose level of the two groups, the stability of blood glucose in the insulin degludec group was better than that in the insulin glargine group. After 12 weeks of treatment, 8 cases (8/15) in insulin degludec group had HbA 1c<7.0%, and 4 cases (4/15) in insulin glargine group had HbA 1c<7.0%, without statistically significant difference ( P=0.264). There were 7 cases (7/15) in the insulin degludec group and 1 case (1/15) in the insulin glargine group who achieved high quality blood glucose control, with statistically significant difference ( P=0.035). At the 12th week of outpatient follow-up, the incidence of nocturnal hypoglycemic events in insulin degludec group was significantly lower than that in insulin glargine group (4/15 vs 11/15, P=0.027). Conclusions:Compared with insulin glargine, insulin degludec can achieve higher blood glucose compliance rate, lower blood glucose level and reduce blood glucose fluctuations in patients with type 1 diabetes.

Objective:A mouse model of pancreatitis induced by coxsackievirus B3 (CVB3) was established. The pathological change of pancreas and the infiltration of Th17/Treg cells were observed.Methods:The BALB/c mice were inoculated intraperitoneally with CVB3 to induce acute viral pancreatitis model. Then the pathological changes of pancreas were observed by HE staining; the viral RNA load and relative expression of cytokines (IFN-γ, IL-6 and IL-17) mRNA were detected by q-PCR; the proportion of infiltrated CD45 + CD3 + T cells, CD4 + and CD8 + T cells, Th17 and Treg cells in the pancreas was determined by flow cytometry. Results:Three days after CVB3 infection, the viral RNA load in pancreas was the highest (0.96±0.18) and gradually decreased with prolongation of infection. Compared with the 3 dpi group, the viral RNA load in pancreas was decreased (0.96±0.18 vs. 0.62±0.14) at 7 dpi, but there was no statistically significant difference. In addition, the infiltration of immune cell in pancreas increased significantly after 7dpi and the pathological score >2. The percent of infiltrated Th17 cells (1.05±0.21 vs. 22.13±5.79) and Treg cells (3.11±0.78 vs. 8.25±1.30) among CD4 + T cells significantly increased after infection (P<0.05), and the Th17/Treg also increased (P<0.01). Compared with the control group, the relative mRNA expression of IFN-γ (1.05±0.23 vs. 672.6±47.67), IL-6 (1.00±0.38 vs. 68.28±4.57), and IL-17 (1.01±0.11 vs. 54.15±7.94) in pancreas increased at 7 days after CVB3 infection ( P<0.01). Conclusions:The infiltration of Th17/Treg cells and the expression of related cytokines related cytokines IL-6 and IL-17 mRNA were upregulated in pancreas, which promoted the process of CVB3-induced pancreatitis.

Objective:To analyze the association of respiratory syncytial virus infection with meteorological factors and to predict and explain the trends.Methods:Data of cases with severe acute respiratory infections in hospitalized children in Xuzhou City were collected from 2015-2021. Respiratory syncytial virus (RSV) was detected by real-time fluorescence polymerase chain reaction. The result were statistically analyzed using SPSS 26.0 software, including constructing a negative binomial regression model to explore meteorological factors that impact RSV detection and a multivariate time series model to predict its epidemiological trend from 2020 to 2021.Results:A total of 1 663 samples of children with severe acute respiratory infections were collected from 2015to 2021, of which 218 (13.1%) were positive for RSV. Seasonal effects on RSV detection were evident: there was a 1-year cycle with a peak in winter (December-February) and a trough in summer (June-August). The negative binomial regression analysis showed that monthly mean temperature, monthly mean relative humidity, and monthly total sunshine hours may influenced RSV detection. The prediction result of the time series model with sunshine hours as the covariate showed that the prediction was better for 2020, and the actual values were close to the predicted values. The expected trends in 2021 were consistent, but the actual values were higher than predicted.Conclusions:Monthly mean temperature, monthly mean relative humidity, and monthly total sunshine hours may influence RSV detection in the Xuzhou region.A prediction model can be built using data from 2015-2019, where deviations in the predicted values for 2021, reflecting that disease prevalence is multifactorial correlated, suggest a possible rise in RSV prevalence in the future.