Small extracellular vesicles facilitate epithelial-mesenchymal transition in chronic rhinosinusitis with nasal polyps via the miR-375-3p/QKI axis.

BACKGROUND: Epithelial-mesenchymal transition (EMT) plays a crucial role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). However, the involvement of small extracellular vesicles (sEVs) in EMT and their contributions to CRSwNP has not been extensively investigated. METHODS: SEVs were isolated from nasal mucosa through ultracentrifugation. MicroRNA sequencing and reverse-transcription quantitative polymerase chain reaction were employed to analyze the differential expression of microRNAs carried by sEVs. Human nasal epithelial cells (hNECs) were used to assess the EMT-inducing effect of sEVs/microRNAs. EMT-associated markers were detected by western blotting and immunofluorescence. Dual-luciferase reporter assay was performed to determine the target gene of miR-375-3p. MicroRNA mimic, lentiviral, and plasmid transduction were used for functional experiments. RESULTS: In line with the greater EMT status in eosinophilic CRSwNP (ENP), sEVs derived from ENP (ENP-sEVs) could induce EMT in hNECs. MiR-375-3p was elevated in ENP-sEVs compared to that in control and nonENP. MiR-375-3p carried by ENP-sEVs facilitated EMT by directly targeting KH domain containing RNA binding (QKI) at seed sequences of 913-919, 1025-1033, and 2438-2444 in 3’-untranslated region. Inhibition of QKI by miR-375-3p overexpression promoted EMT, which could be reversed by restoration of QKI. Furthermore, the abundance of miR-375-3p in sEVs was closely correlated with the clinical symptom score and disease severity. CONCLUSIONS: MiR-375-3p-enriched sEVs facilitated EMT by suppressing QKI in hNECs. The association of miR-375-3p with disease severity underscores its potential as both a diagnostic marker and a therapeutic target for the innovative management of CRSwNP.

[Factors influencing primary patency time of percutaneous transluminal balloon angioplasty for hemodialysis arteriovenous grafts].

Objective: To investigate the factors affecting primary patency time in arteriovenous graft (AVG) patients receiving percutaneous transluminal balloon angioplasty (PTA). Methods: Hemodialysis patients who underwent AVG placement at the First Affiliated Hospital of Chongqing Medical University between February 2018 and December 2021 were included. The factors including age, gender, total duration of AVG use, site of stenosis, degree of stenosis, length of stenosis, residual stenosis, and presence of thrombosis were analyzed, and influencing factors of primary patency time in AVG were determined using a multiple linear regression model. Results: A total of 101 patients who underwent 331 PTA treatments were enrolled, including 35 males and 66 females. The median age of patients undergoing PTA for the first time was 61 (51, 68) years, and the primary patency time after PTA was 5 (3, 10) months. The patients were followed up for (38.5±15.3) months. Multivariable linear regression analysis revealed that severe stenosis at the venous anastomosis and reflux veins (ß=-2.773, 95%CI:-5.440--0.105, P=0.042), female (ß=-2.247, 95%CI:-3.853--0.642, P=0.006), and previous multiple PTA treatments (ß=-0.516, 95%CI:-0.978--0.054, P=0.029) were risk factors for a shorter primary patency time after PTA. Conclusion: Severity of stenosis at the venous anastomosis and reflux veins of the AVG, female, and a history of multiple previous PTA treatments are associated with a shorter primary patency time in AVG patients.

[Current status and reflection on molecular diagnosis of birth defects].

Birth defects are a major problem threatening the health of children in China. Genetic factors play a major role in birth defect etiology. Molecular diagnosis is the key means for screening, diagnosing, and preventing birth defects caused by genetic factors. How to carry out large-scale and cost-effective molecular diagnosis in clinical practice is a major challenge in the prevention and treatment of birth defects in China. This article reviews the current status of birth defects in China, the application of molecular diagnostic technology in birth defect prevention and control, and the challenges in promoting its use, to provide references for clinical practice in birth defect molecular diagnosis.

[Analysis of pollen sensitization characteristics of artemisia allergic rhinitis in three urban and rural areas of Inner Mongolia].

Objective: To investigate the distribution rules of artemisia pollen and the clinical sensitization characteristics of allergic rhinitis (AR) induced by artemisia pollen in three urban and rural areas of Inner Mongolia. Methods: From March to October 2019, in 3 central cities (Chifeng, Hohhot, Ordos) and rural areas of Inner Mongolia, an epidemiological investigation method combining multi-stage stratified random sampling and face-to-face questionnaire survey was adopted to screen suspected AR patients, and skin prick test (SPT) was applied for diagnosis. At the same time, pollen monitoring was carried out in 3 areas to analyze the distribution and clinical sensitization characteristics of artemisia pollen.SPSS26.0 statistical software was used to process all the data. Chi-square test was used to compare rates among different age, sex, region and nationality, Spearman test was used to describe correlation analysis, and pairwise comparison of positive rates among multiple samples was used Bonferroni method. Results: Among the 6 393 subjects, 1 093 cases were diagnosed with AR, and the prevalence of AR was 17.10% (1 093/6 393). Among them, pollen-induced allergic rhinitis, the prevalence of PiAR was 10.97% (701/6 393), accounting for 64.14%(701/1 093).The highest incidence was in the youth group (20-39 years old), accounting for 46.94% (329/701).The diagnosed prevalence was higher in females than in males (11.35% vs. 10.64%, χ2 value 12.304, P<0.001).The prevalence rate of ethnic minority was higher than that of Han nationality (13.01% vs. 10.65%, χ2 value 6.296, P=0.008).The prevalence in urban areas was also significantly higher than that in rural areas (18.40% vs. 5.50%, χ2 value 10.497, P<0.001).There was significant difference in prevalence rate among the three regions in Inner Mongolia (6.06% in Chifeng, 13.46% in Hohhot, 16.39% in Ordos, χ2 value 70.054, P<0.001).The main clinical symptoms of artemisia PiAR were sneezing (95.58%), nasal congestion (91.73%) and nasal itching (89.30%).Allergic conjunctivitis accounted for 79.60% (558/701), chronic sinusitis for 55.63% (390/701), asthma for 23.25% (163/701).The pattern of artemisia pollen sensitization was mainly multiple sensitization, and the frequency of clinical symptoms and clinical diseases induced by hypersensitization with other allergens accounted for more than that caused by single artemisia pollen. The spread period of Artemisia pollen in the three regions was from June to October, and the peak state was in August in summer. The peak time of clinical symptoms in artemisia PiAR patients was about 2 weeks earlier than the peak time of pollen concentration, and the two were significantly positively correlated (R=0.7671, P<0.001). Conclusion: Artemisia pollens are the dominant pollens in late summer and early autumn in Inner Mongolia, and the prevalence of artemisia PiAR is high. Controlling the spread of Artemisia pollens is of great significance for the prevention and treatment of AR.

Salivary Microbiome Relates to Neoadjuvant Immunotherapy Response in OSCC.

Most patients diagnosed with oral squamous cell carcinoma (OSCC) present with locally advanced stages, which are typically associated with poor outcomes. Although immunotherapy offers potential improvements in patient survival, its efficacy is hampered by low response rates. The microbiome is widely involved in tumor immunity and may play a role in immunotherapy. This study aimed to investigate the potential association between the oral (salivary) microbiome and immunotherapy response in patients with OSCC. Salivary metagenome sequencing was performed on 47 patients with OSCC undergoing neoadjuvant immunotherapy (NAIT) in a clinical trial (NCT04649476). Patients were divided into responders and nonresponders based on their pathological responses. The results showed that the species richness of the salivary microbiome was lower in the nonresponders before NAIT than in the responders. Differential analysis revealed that nonresponders exhibited a lower relative abundance of 34 bacterial species and a higher relative abundance of 4 bacterial species. Notably, low levels of Eubacterium infirmum, Actinobaculum, and Selenomas (EAS) in the saliva may be associated with the nonresponse of patients with OSCC to NAIT. A nomogram based on EAS was developed and validated to determine the efficacy of NAIT. The area under the curve for the training cohort was 0.81 (95% confidence interval, 0.66 to 0.81). Quantitative polymerase chain reaction confirmed that low levels of salivary EAS effectively identified nonresponders to NAIT. Furthermore, the low abundance of salivary EAS was closely correlated with a low density of intratumoral CD4+, CD14+, CD68+, and FOXP3+ cells. Metabolic functional annotation revealed numerous biosynthetic processes associated with EAS that were more active in responders. In summary, this study provides valuable data resources for the salivary microbiome and reveals that nonresponders have different salivary microbiome profiles than responders do before NAIT. Low salivary EAS levels can serve as potential biomarkers for distinguishing nonresponders from responders.

[Application effects of rehabilitation care decision-making scheme based on case management model in severe burn patients].

Objective: To explore the application effects of application of rehabilitation care decision-making scheme based on case management model in severe burn patients. Methods: The study was a non-randomized historical control study. Thirty patients who met the inclusion criteria and received routine rehabilitation nursing in the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the hospital) from April 2021 to March 2022 were included in routine rehabilitation nursing group (26 males and 4 females, aged 48.50 (31.75, 56.25) years), and 30 patients who met the inclusion criteria and received case management rehabilitation nursing in the hospital from April 2022 to March 2023 were included in case management rehabilitation nursing group (22 males and 8 females, aged 46.00 (36.75, 55.25) years). The length of intensive care unit (ICU) stay, total hospitalization day, and total hospitalization cost of the patients in two groups were recorded. At admission, convalescence, discharge, and 6 months after injury, the patients' life quality was evaluated by the concise burn specific health scale, the sleep quality was evaluated by the Pittsburgh sleep quality index, and the functional independence was evaluated by the functional independence rating scale. At convalescence, discharge, and 6 months after injury, the patients' scar status was evaluated by the Vancouver scar scale. At 6 months after injury, a third-party satisfaction questionnaire was used to investigate the efficacy satisfaction of patients. Results: The length of ICU stay and total hospitalization day of patients in case management rehabilitation nursing group were both significantly shorter than those in routine rehabilitation nursing group (with Z values of -1.97 and -1.99, respectively, P<0.05), and the total hospitalization cost was less than that in routine rehabilitation nursing group (Z=-1.99, P<0.05). At discharge and 6 months after injury, the life quality scores of patients in case management rehabilitation nursing group were significantly higher than those in routine rehabilitation nursing group (with t values of -3.19 and -4.43, respectively, P<0.05), while the sleep quality scores were significantly lower than those in routine rehabilitation nursing group (with Z values of -2.18 and -3.33, respectively, P<0.05). There were no statistically significant differences in cognitive function scores of functional independence of patients between the 2 groups at admission, convalescence, discharge, and 6 months after injury (P>0.05). The exercise function scores and total scores of functional independence of patients in case management rehabilitation nursing group at convalescence, discharge, and 6 months after injury were significantly higher than those in routine rehabilitation nursing group (with Z values of -4.37, -2.73, -4.10, -4.37, -2.64, and -4.06, respectively, P<0.05). The scar pigmentation scores of patients in case management rehabilitation nursing group at 6 months after injury were significantly lower than those in routine rehabilitation nursing group (Z=-2.05, P<0.05), and the scar vascularity scores of patients in case management rehabilitation nursing group at discharge and 6 months after injury in case management rehabilitation nursing group were significantly lower than those in routine rehabilitation nursing group (with Z values of -3.16 and -2.07, respectively, P<0.05). The scar pliability scores (with Z values of -3.16, -2.45, and -4.38, respectively, P<0.05), thickness scores (with Z values of -2.56, -2.35, and -4.70, respectively, P<0.05), and total scores (with Z values of -3.77, -3.04, and -3.13, respectively, P<0.05) of patients in case management rehabilitation nursing group at convalescence, discharge, and 6 months after injury were significantly lower than those in routine rehabilitation nursing group. At 6 months after injury, the efficacy satisfaction scores of patients in case management rehabilitation nursing group were 4.00 (3.00, 4.25), which were significantly higher than 3.00 (2.00, 4.00) in routine rehabilitation nursing group (Z=-2.72, P<0.05). Conclusions: The implementation of rehabilitation care decision-making scheme based on case management model can optimize the cost efficiency, improve the effectiveness of clinical treatment, and enhance the life quality and satisfaction of the curative effect of severe burn patients.

[Exosomes from ectoderm mesenchymal stem cells inhibits lipopolysaccharide-induced microglial M1 polarization and promotes survival of H2O2-exposed PC12 cells by suppressing inflammatory response and oxidative stress].

OBJECTIVE: To investigate the potential value of exosomes derived from rat ectoderm mesenchymal stem cells (EMSCs-exo) for repairing secondary spinal cord injury. METHODS: EMSCs-exo were obtained using ultracentrifugation from EMSCs isolated from rat nasal mucosa, identified by transmission electron microscope, nanoparticle tracking analysis (NTA), and Western blotting, and quantified using the BCA method. Neonatal rat microglia purified by differential attachment were induced with 100 µg/L lipopolysaccharide (LPS) and treated with 37.5 or 75 mg/L EMSCs-exo. PC12 cells were exposed to 400 µmol/L H2O2 and treated with EMSCs-exo at 37.5 or 75 mg/L. The protein and mRNA expressions of Arg1 and iNOS in the treated cells were determined with Western blotting and qRT- PCR, and the concentrations of IL- 6, IL-10, and IGF-1 in the supernatants were measured with ELISA. The viability and apoptosis of PC12 cells were detected using CCK-8 assay and flow cytometry. RESULTS: The isolated rat EMSCs showed high expressions of nestin, CD44, CD105, and vimentin. The obtained EMSCs-exo had a typical cup-shaped structure under transmission electron microscope with an average particle size of 142 nm and positivity for CD63, CD81, and TSG101 but not vimentin. In LPS-treated microglia, EMSCs-exo treatment at 75 mg/L significantly increased Arg1 protein level and lowered iNOS protein expression (P < 0.05). EMSCs-exo treatment at 75 mg/L, as compared with the lower concentration at 37.5 mg/L, more strongly increased Arg1 mRNA expression and IGF-1 and IL-10 production and decreased iNOS mRNA expression and IL-6 production in LPS-induced microglia, and more effectively promoted cell survival and decreased apoptosis rate of H2O2-induced PC12 cells (P < 0.05). CONCLUSION: EMSCs-exo at 75 mg/L can effectively reduce the proportion of M1 microglia and alleviate neuronal apoptosis under oxidative stress to promote neuronal survival, suggesting its potential in controlling secondary spinal cord injury.

[Inhibition of glutamatergic neurons in the dorsomedial periaqueductal gray alleviates excessive defensive behaviors of mice with post-traumatic stress disorder].

OBJECTIVE: To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey (dmPAG) in regulating excessive defensive behaviors in mice with post-traumatic stress disorder (PTSD). METHODS: Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno- associated viral vectors (rAAV2/9-CaMKII-mCherry, rAAV2/9-CaMKII-hM3Dq-mCherry and rAAV2/9-CaMKII-hM4Di-mCherry) into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons, followed 2 weeks later by PTSD modeling by single prolonged stress. The looming test, response to whisker stimulation test and contextual fear conditioning (CFC) test were used to observe changes in defensive behaviors of the PTSD mice. The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining. RESULTS: Compared with the control mice, the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest, response scores of defensive behaviors and freezing time (all P<0.01). Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors. Activation of the glutamatergic neurons in the dmPAG (in PTSD hM3Dq group) did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice, whereas inhibiting the glutamatergic neurons in the dmPAG (in PTSD hM4Di group) caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice (P<0.05 or 0.01). CONCLUSION: Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.