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.

[Clinical characteristics of hospitalized children with respiratory syncytial virus infection and risk prediction of severe illness during the post-COVID-19 era in Kunming].

Objective: To compare the epidemiological and clinical characteristics of hospitalized children with respiratory syncytial virus (RSV) infection in Kunming among the pre-and post-COVID-19 era, and to establish a prediction model for severe RSV infection in children during the post-COVID-19 period. Methods: This was a retrospective study. Clinical and laboratory data were collected from 959 children hospitalized with RSV infection in the Department of Pulmonary and Critical Care Medicine at Kunming Children's Hospital during January to December 2019 and January to December 2023. Patients admitted in 2019 were defined as the pre-COVID-19 group, while those admitted in 2023 were classified as the post-COVID-19 group. Epidemiological and clinical characteristics were compared between the two groups. Subsequently, comparison of the clinical severity among the two groups was performed based on propensity score matching (PSM). Furthermore, the subjects in the post-COVID-19 group were divided into severe and non-severe groups based on clinical severity. Chi-square test and Mann-Whitney U test were used for pairwise comparison between groups, and multivariate Logistic regression was applied for the identification of independent risk factors and construction of the prediction model. The receiver operating characteristic (ROC) curve and calibration curve were employed to evaluate the predictive performance of this model. Results: Among the 959 children hospitalized with RSV infection, there were 555 males and 404 females, with an onset age of 15.4 (7.3, 28.5) months. Of which, there were 331 cases in the pre-COVID-19 group and 628 cases in the post-COVID-19 group. The peak period of RSV hospitalization in the post-COVID-19 group were from May to October 2023, and the monthly number of inpatients for each of these months were as follows: 72 cases (11.5%), 98 cases (15.6%), 128 cases (20.4%), 101 cases (16.1%), 65 cases (10.4%), and 61 cases (9.7%), respectively. After PSM for general data, 267 cases were matched in each group. The proportion of wheezing in the post-COVID-19 group was lower than that in the pre-COVID-19 group (109 cases (40.8%) vs. 161 cases (60.3%), χ2=20.26, P<0.001), while the incidences of fever, tachypnea, seizures, severe case, neutrophil-to-lymphocyte ratio (NLR), C-reactive protein and interleukin-6 levels were all higher than those in the pre-COVID-19 group (146 cases (54.7%) vs. 119 cases (44.6%), 117 cases (43.8%) vs. 89 cases (33.3%), 37 cases (13.9%) vs. 14 cases (5.2%), 69 cases (25.8%) vs. 45 cases (16.9%), 3.6 (1.9, 6.4) vs. 2.3 (1.8, 4.6), 9.9 (7.1, 15.2) vs. 7.8 (4.5, 13.9) mg/L, 20.5 (15.7, 30.4) vs. 17.2 (11.0, 26.9) ng/L, χ2=5.46, 6.36, 11.47, 6.42, Z=4.13, 3.06, 2.96, all P<0.05). There were 252 cases and 107 cases with co-infection in the post-and pre-COVID-19 groups, respectively. The proportion of triple and quadruple infection in the post-COVID-19 group was higher than that in the pre-COVID-19 group (59 cases (23.4%) vs. 13 cases (12.1%), 30 cases (11.9%) vs. 5 cases (4.7%), χ2=5.94, 4.46, both P<0.05). Among the 252 cases with co-infection in post-COVID-19 group, the most prevalent pathogens involving in co-infections, in order, were Mycoplasma pneumoniae 56 cases (22.2%), Influenza A virus 53 cases (21.0%), Rhinovirus 48 cases (19.0%), Parainfluenza virus 35 cases (13.9%), and Adenovirus 28 cases (11.1%).The result of multivariate Logistic regression showed that age (OR=0.70, 95%CI 0.62-0.78, P<0.001), underlying diseases (OR=10.03, 95%CI 4.10-24.55, P<0.001), premature birth (OR=6.78, 95%CI 3.53-13.04, P<0.001), NLR (OR=1.85, 95%CI 1.09-3.15, P=0.023), and co-infection (OR=1.28, 95%CI 1.18-1.38, P<0.001) were independently associated with the development of severe RSV infection in the post-COVID-19 group. The ROC curve of the prediction model integrating the above five factors indicated an area under the curve of 0.85 (95%CI 0.80-0.89, P<0.001), with an optimal cutoff of 0.21, a sensitivity of 0.83 and a specificity of 0.80. The calibration curve showed that the predicted probability in this model did not differ significantly from the actual probability (P=0.319). Conclusions: In the post-COVID-19 era in Kunming, the peak in pediatric hospitalizations for RSV infection was from May to October, with declined incidence of wheezing and increased incidence of fever, tachypnea, seizures, severe cases, and rates of triple and quadruple co-infections. Age, underlying diseases, premature birth, NLR, and co-infection were identified as independent risk factors for severe RSV infection in the post-COVID-19 period. In this study, a risk prediction model for severe pediatric RSV infection was established, which had a good predictive performance.

Body mass index’s effect on CRSwNP extends to pathological endotype and recurrence.

BACKGROUND: Elevated body mass index (BMI) has been recognized as an important contributor to corticosteroid insensitivity in chronic rhinosinusitis with nasal polyps (CRSwNP). We aimed to delineate the effects of elevated BMI on immunological endotype and recurrence in CRSwNP individuals. METHODOLOGY: A total of 325 patients with CRSwNP undergoing FESS were recruited and stratified by BMI. H&E staining was employed for histological evaluation. Characteristics of inflammatory patterns were identified by immunohistochemical staining. The predictive factors for recurrence were determined and evaluated by multivariable logistic regression analysis and the receiver operating characteristic (ROC) curves across all subjects and by weight group. RESULTS: In all patients with CRSwNP, 26.15% subjects were classified as overweight/obese group across BMI categories and exhibited a higher symptom burden. The upregulated eosinophil/neutrophil-dominant cellular endotype and amplified type 2/ type 3 coexisting inflammation was present in overweight/obese compared to underweight/normal weight controls. Additionally, a higher recurrent proportion was shown in overweight/obese patients than that in underweight/normal weight cohorts. Multivariable logistic regression analysis identified BMI as an independent predictor for recurrence. The predictive capacity of each conventional parameter (tissue eosinophil and CLCs count, and blood eosinophil percentage) alone or in combination was poor in overweight/obese subjects. CONCLUSIONS: Overweight/obese CRSwNP stands for a unique phenotype and endotype. Conventional parameters predicting recurrence are compromised in overweight/obese CRSwNP, and there is an urgent need for novel biomarkers that predict recurrence for these patients.

[Clinical characteristics of children with SARS-CoV-2 Omicron variant infection in Kunming].

Objective: To investigate the clinical characteristics of hospitalized children infected with the Omicron variant in Kunming after the withdrawal of non-pharmaceutical interventions (NPI) and analyze the risk factors of severe cases. Methods: Clinical data was retrospectively collected from 1 145 children with SARS-CoV-2 Omicron infection who were hospitalized in six tertiary grade A hospitals in Kunming from December 10th, 2022 to January 9th, 2023. According to clinical severity, these patients were divided into the general and severe SARS-CoV-2 groups, and their clinical and laboratory data were compared. Between-group comparison was performed using t-test, chi-square test and Mann-Whitney U test. Spearman correlation test and multivariate Logistic regression analysis were used to determine the risk factors of severe illness. Results: A total of 1 145 hospitalized patients were included, of whom 677 were male and 468 female. The age of these patients at visit was 1.7 (0.5, 4.1) years. Specifically, there were 758 patients (66.2%) aged ≤3 years at visit and 387 patients (33.8%) aged >3 years. Of these children, 89 cases (7.8%) had underline diseases and the remaining 1 056 cases (92.2%) had no combined diseases. Additionally, of all the patients, 319 cases (27.9%) were vaccinated with one or two doses of SARS-CoV-2 vaccine, 748 cases (65.3%) had acute upper respiratory tract infection (AURTI), and six cases died (0.5%). A total of 1 051 cases (91.8%) were grouped into general SARS-CoV-2 group and 94 cases (8.2%) were grouped into severe SARS-CoV-2 group. Compared with the general cases, the severe cases showed a lower rate of SARS-CoV-2 vaccination and younger median age, lower lymphocyte count, as well as proportions of CD8+T lymphocyte (36 cases (38.3%) vs. 283 cases (26.9%), 0.5 (2.6, 8.0) vs. 1.6 (0.5, 3.9) years, 1.3 (1.0, 2.7) ×109 vs. 2.7 (1.3,4.4)×109/L, 0.17 (0.12, 0.24) vs. 0.21 (0.15, 0.16), respectively, χ2=4.88, Z=-2.21,-5.03,-2.53, all P<0.05). On the other hand, the length of hospital stay, proportion of underline diseases, ALT, AST, creatine kinase isoenzyme, and troponin T were higher in the severe group compared to those in the general group ((11.6±5.9) vs. (5.3±1.8) d, 41 cases (43.6%) vs. 48 cases (4.6%), 67 (26,120) vs. 20 (15, 32) U/L, 51 (33, 123) vs. 44 (34, 58) U/L、56.9 (23.0, 219.3) vs. 3.6 (1.9, 17.9) U/L, 12.0 (4.9, 56.5) vs. 3.0 (3.0, 7.0) ×10-3 pg/L,respectively, t=-20.43, χ2=183.52, Z=-9.14,-3.12,-6.38,-3.81, all P<0.05). Multivariate regression analysis indicated that increased leukocyte count (OR=1.88, 95%CI 1.18-2.97, P<0.01), CRP (OR=1.18, 95%CI 1.06-1.31, P<0.01), ferritin (OR=1.01, 95%CI 1.00-1.00, P<0.01), interleukin (IL)-6 (OR=1.05, 95%CI 1.01-1.08, P=0.012), D-dimer (OR=2.56, 95%CI 1.44-4.56, P<0.01) and decreased CD4+T lymphocyte (OR=0.84, 95%CI 0.73-0.98, P=0.030) were independently associated with the risk of severe SARS-CoV-2 in hospitalized children with Omicron infection. Conclusions: After the withdrawal of NPI, the pediatric inpatients with Omicron infection in Kunming were predominantly children younger than 3 years of age, and mainly manifested as AURTI with relatively low rate of severe SARS-CoV-2 infection and mortality. Elevated leukocyte counts, CRP, ferritin, IL-6, D-dimer, and decreased CD4+T lymphocytes are significant risk factors for developing severe SARS-CoV-2 infection.

A feasibility study on the efficacy of a patient-owned wound surveillance system for diabetic foot ulcer care (ePOWS study).

Objective: Wound image analysis tools hold promise in helping patients to monitor their wounds. We aim to perform a novel feasibility study on the efficacy of a patient-owned wound surveillance system for diabetic foot ulcer (DFU) care. Methods: This two-institutional, prospective, single-arm pilot study examined patients with DFU. An artificial intelligence-enabled image analysis app calculating the wound surface area was installed and patients or caregivers were instructed to take pictures of wounds during dressing changes. Patients were followed until wound deterioration, wound healing, or wound stability at 6 months occurred and the outcomes of interest included study adherence, algorithm performance, and user experience. Results: Between January 2021 and December 2021, 39 patients were enrolled in the study, with a mean age of 61.6 ± 8.6 years, and 69% (n = 27) of subjects were male. All patients had documented diabetes and 85% (n = 33) of them had peripheral arterial disease. A mean follow-up for those completing the study was 12.0 ± 8.5 weeks. At the conclusion of the study, 80% of patients (n = 20) had primary wound healing whilst 20% (n = 5) had wound deterioration. The study completion rate was 64% (n = 25). Usage of the app for surveillance of DFU healing, as compared to physician evaluation, yielded a sensitivity of 100%, specificity of 20%, positive predictive value of 83%, and negative predictive value of 100%. Of those who provided user experience feedback, 59% (n = 10) felt the app was easy to use, 47% (n = 8) would recommend the wound analysis app to others but only 6% would pay for the app out of pocket (n = 1). Conclusion: Implementation of a patient-owned wound surveillance system is feasible. Most patients were able to effectively monitor wounds using a smartphone app-based solution. The image analysis algorithm demonstrates strong performance in identifying wound healing and is capable of detecting deterioration prior to interval evaluation by a physician. Patients generally found the app easy to use but were reluctant to pay for the use of the solution out of pocket.

[Establishment and behavioral evaluation of a mouse model of long-term free-choice alcohol drinking].

OBJECTIVE: To establish a model of long-term free drinking mouse by feeding mice with alcohol to simulate the state of human voluntary long-term drinking, and on this basis, to further discuss the evaluation criteria of long-term free drinking mice model in sports, anxiety and cognitive behavior. METHODS: Forty six-week-old SPF C57BL/6 male mouse were randomly divided into two groups: Long-term free drinking group (n=20) and normal control group (n=20). The two groups were given solid feed normally. The long-term free drinking group was free to take 10% alcohol and water every day, while the normal drinking group only took water every day. The mice were fed for 7 months, and were evaluated by a series of behavioral methods, including Rota-rod test, balance beam test, open filed test, the elevated plus maze, two-box social behavior, new object recognition, Y maze and water maze. RESULTS: With the increase of drinking days, the mice showed significant alcohol addiction in the alcohol preference test. With the increase of alcohol intake, the mice in the long-term free choice drinking group had slightly shiny fur and reduced diet. Compared with the control group, the weight gain began to slow down from the third month, and the weight decreased significantly by the sixth and seventh months (P=0.006, P < 0.001). The mice showed reduced balance locomotion ability (P=0.003, P=0.001) in the rotary bar and balance beam test. In the open field and elevated cross test, the mice had obvious anxiety-like behavior (P < 0.001). The mice showed decreased social ability in the two boxes of social behavior (P < 0.016). In the experiment of new object recognition and Y maze, the exploration of new object decreased (P=0.018, P=0.040). In the water maze, cognitive functions, such as learning and spatial memory were reduced (P < 0.001). CONCLUSION: The successful establishment of the long-term free drinking mouse model is more convenient for us to carry out further research on the neural mechanism of alcohol addiction, and lays an experimental foundation for exploring the neural mechanism of alcohol addiction and related new targets.

[Correlation between contrast-enhanced ultrasound parameters and Crohn's disease activity].

Objective: By investigating the correlation between quantitative parameters of contrast enhanced ultrasound (CEUS) and commonly used activity assessment indicators of Crohn's disease (CD), and comparing the predictive power of laboratory inflammatory indicators with CEUS on Crohn's disease (CD), the significance of CEUS was evaluated. Methods: A case-control study. From October 2019 to December 2021, the clinical data of 67 patients with CD who were diagnosed by endoscopy and underwent contrast-enhanced ultrasonography were retrospectively analyzed in the First Affiliated Hospital with Nanjing Medical University, and their routine ultrasound and CEUS parameters, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), fecal calprotectin (FC), Crohn's disease activity index (CDAI) and simplified endoscopic score for Crohn's disease (SES-CD) were collected. Using SES-CD as the standard, the patients were divided into a remission group and an active group, and the correlation of laboratory inflammatory indexes and contrast-enhanced ultrasound parameters with CDAI and SES-CD were evaluated. Besides, the ROC curve was used to analyze the predictive efficacy of each index on CD endoscopic activity. Results: A total of 67 patients were included in this study. According to the SES-CD score, there were 17 patients in the remission group and 50 patients in the active group. Except for the coefficient of the enhancement wash in slope and time to peak (TTP), the peak intensity (PI), area under the angiography curve, and laboratory inflammatory indexes were significantly different between the two groups (P<0.05), which also showed a moderate positive correlation with CDAI and SES-CD (P<0.05). ROC analysis showed that among the non-invasive indicators, PI and area under the angiography curve had the highest AUCs for predicting CD endoscopic activity, which were 0.912 and 0.891, respectively; with SES-CD taking >3 as the cut-off value, the corresponding sensitivities were 78.0% and 72.0%, with specificities of 100.0% and 94.1%, respectively. Conclusion: CEUS can objectively and repeatedly evaluate the disease activity of CD patients, and has great clinical application value, which can be used as a reliable imaging method for diagnosis and follow-up of patients with Crohn's disease.

[Application of locating supratrochlear artery and supraorbital artery in combined transfrontal and intranasal endoscopic approaches].

Objective: To investigate the localization methods of supratrochlear artery (STA) and supraorbital artery (SOA), and to explore the clinical benefit of locating nerve via accompanying vascular localization in combined transfrontal and intranasal endoscopic approaches. Methods: From June 2019 to May 2021, 14 patients, including 11 males and 3 females, aging from 18 to 69 years old, were underwent frontal sinus surgery through the combined transfrontal and intranasal endoscopic approaches in the Department of Otorhinolaryngology Head and Neck Surgery of the Third Affiliated Hospital of Sun Yat-sen University. Before the surgery, localization of STA and SOA was determined by color doppler flow imaging (CDFI), computerized topographic angiography (CTA) and contrast enhanced magnetic resonance angiography (CE-MRA) respectively, and the distances between STA and SOA from facial midline were measured on 28 eyebrows. The position of external incision was determined according to the preoperative localization of STA and SOA. The examination time, cost and postoperative complications of the three methods were recorded. The accuracy of localization at 14 sides was verified by the surgery. GraphPad Prism 8.3 software was used for statistical analysis. Results: STA and SOA could be located by CDFI, CTA and CE-MRA. There was no significant difference in the measurement of the distance between STA and SOA from the facial midline among 3 methods (all P>0.05). Determining the position of external incision according to the localization of STA and SOA could protect both the blood vessels and accompanying nerves. No postoperative complications such as numbness of the forehead skin occurred. The measurement time of CDFI, CTA and CE-MRA was 22.50 (15.75, 30.00), 30.00 (28.00, 34.25) and 48.00 (44.00, 52.75) min (M (Q1, Q3)), respectively (all P<0.05). CDFI incurred the lowest costs and took the shortest time. Conclusions: CDFI is an efficient and economic localization method. The localization of STA and SOA facilitates the precise selection of the position of external incision, protects the accompanying nerve and reduces postoperative complications.

The cross-sectional study of depressive symptoms and associated factors among adolescents by backpropagation neural network.

OBJECTIVES: This study aimed to investigate the association between depressive symptoms and diet- and lifestyle-related behaviors among adolescents. STUDY DESIGN: Cross-sectional study. METHODS: Our study used stratified random cluster sampling method to recruit 6,251 adolescents aged 11-19 years as samples for research and analysis. The Center for Epidemiological Studies Depression Scale was used to assess depressive symptoms. Chi-squared test, t test, and logistic regression were used to explore the diet and lifestyle factors of depressive symptoms. Backpropagation (BP) neural network model was used to investigate the ranking of diet and lifestyle behaviors factors of depressive symptoms. RESULTS: The prevalence of depressive symptoms among adolescents was 32.1%. Multivariable logistic regression was used to determine 10 important variables of depressive symptoms. After ranking the importance by BP neural network, the top three important variables were found, which were sleep duration (100%), screen time (49.1%), and breakfast (23.6%). CONCLUSION: Sleep duration, screen time, and breakfast were associated factors with the most significant impacts on depressive symptoms.

[Pathological characteristics and clinical prognosis of nodular sclerosis grade 2 of classic Hodgkin's lymphoma].

Objective: To investigate the pathological characteristics and clinical prognosis of nodular sclerosis grade 2 of classic Hodgkin's lymphoma (cHL-NS2) in our cancer center. Methods: A retrospective collection of 23 cases of cHL-NS2 admitted in Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from July 2008 to April 2019 was performed. Fifty-five cases of nodular sclerosis grade 1 of classical Hodgkin's lymphoma (cHL-NS1) during the same period were selected as control group. Survival curves were plotted using the Kaplan-Meier method, and Cox regression model was used to analyze the influencing factors for survival. Results: The median age of 23 cases of cHL-NS2 was 30 years old. Five cases had extra nodal invasion, and 19 cases were Ⅰ-Ⅱ stage based on Ann Arbor system. The pathological morphology of cHL-NS2 showed that the lymph node structure was completely destroyed and was divided into nodules by thick collagen. The tumor cells in the nodules were abundant and proliferated in sheets. The boundaries between the tumor cells were not clear. The incidence of tumor necrosis in cHL-NS2 was 43.5% (10/23), which was significantly higher than 18.2% (10/55) in cHL-NS1 (P=0.040). The 3-year progression-free survival (PFS) rate of patients in the cHL-NS2 group was 58.1%, which was significantly lower than 89.7% in the cHL-NS1 group (P=0.002). In all of 78 cases, the 3-year PFS rate of patients who did not obtain complete response (CR) was 67.1%, which was significantly lower than 92.2% in patients who achieved CR (P=0.030). Multivariate Cox regression analysis demonstrated that both cHL-NS2 and failure to obtain CR by first-line treatment were independent indicators for short PFS time (P<0.05). Conclusions: In cHL-NS2, the morphology of tumor cells are diverse, and tumor necrosis can be easily found. Under the current first-line treatments of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) or bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPP), cHL-NS2 is an independent indicator for worse PFS.

Nanocomposite of Ag nanoparticles and deep eutectic solvent-derived carbon dots with oxidase mimicking activity as synergistic bactericidal agent.

A new type of nitrogen and chloride co-doped carbon dots (N/Cl-CDs) based on choline chloride-urea-glycine ternary deep eutectic solvents (DESs) was synthesized using a one-step hydrothermal method. The prepared N/Cl-CDs exhibited oxidase-like activity and excellent antibacterial activity against Escherichia coli, Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA). The addition of silver nanoparticles (Ag NPs) (i.e. N/Cl-CDs + Ag NPs) to the N/Cl-CDs also significantly enhanced the oxidase and antibacterial activities. The nanocomposite (1·8 mg ml-1 ) completely inactivated 105 CFU per ml of MRSA in 90 min. E. coli and S. aureus were labelled with the N/Cl-CDs, enabling multicolour fluorescence imaging at different excitation wavelengths. The nanocomposites have high antibacterial efficiency as a new bactericidal agent, as well as application potential with good biocompatibility and low toxicity.

Change of zinc mobilization and gene expression of key zinc transport proteins between the yolk sac membrane and liver of duck embryonic developing.

Zinc (Zn) deposition in egg yolk is essential for the rapid growth and complete development of the avian embryo. Thus, it is crucial to obtain maximal Zn mobilization at an appropriate time during development in favor of the survival of avian embryos. The aim of this study was to study the developmental change of Zn mobilization and gene expression related to key Zn transport proteins between the yolk sac membrane and embryonic liver from the incubation d 17 (E17) to d 32 (E32) during duck embryonic developing. The weights of duck embryo, embryo without yolk sac, and embryonic liver increased as well as the yolk sac weight decreased linearly (P < 0.0001) when incubation day increased. The Zn concentration in the yolk sac did not change from E17 to E29 and only declined significantly from E29 to E32 of duck embryos, while hepatic Zn level decreased linearly as with the increased incubation time (P < 0.01). When the incubation day increased, the decreased Zn amount in the yolk sac and the increased Zn amount in the embryonic liver were observed (P < 0.0001). The calculated transfer-out rate of Zn in the yolk sac and transfer-in rate of Zn in livers were both increased from E23-26 to E29-32 (P < 0.01). Among E17, E23 and E29, the solute carrier family 39 member (ZIP) of ZIP10, ZIP13, and ZIP14 genes mRNA expressions were increased in yolk sac membrane but were decreased in the embryonic liver, while metallothionein 1 mRNA expression was increased both in the yolk sac membrane and liver (P < 0.05). In conclusion, yolk sac membrane and embryonic liver tissues displayed the similar developmental patterns of Zn mobilization and metallothionein 1 mRNA expression from E17 to E32 during duck embryonic developing. The appropriate time of the maximal rate of Zn mobilization were observed between E29 and E32 of duck embryo, associated with the significant changes of gene expression related to some key Zn transport proteins on E29 in yolk sac membrane and liver tissues.

[Effects of glucocorticoids on the proliferation of CD19 CAR-T cells targeting B-cell tumor cell lines].

Objective: To evaluate the effects of glucocorticoids (dexamethasone and methylprednisolone) on the proliferation of CD19 Chimeric antigen receptor (CAR) modified T cells in vitro. Methods: Peripheral blood mononuclear cells from healthy volunteers were collected as T cells. CD19 CAR-T cells were prepared by CD3 magnetic beads sorting and CD19 CAR lentivirus transfection. The transfection rates and the proportion of CD19 CAR-T cells in the culture system were analyzed using a flow cytometer. The mean fluorescence intensity (MFI) of CD19 CAR-T cells was measured after staining with Carboxyfluorescein diacetate succinimidyl ester cell proliferation tracer fluorescent probe, Lactate dehydrogenase (LDH) cytotoxicity assay was used to detect the effects of different concentrations of glucocorticoid on the killing activity of B-cell tumor cell lines. Results: In this study, the CD19 CAR transfection rate of CD19 CAR-T cells was (51.34±5.28) %. The killing activities of different doses of methylprednisolone on Nalm6, Pfeiffer, and U2932 tumor cells were higher than that of dexamethasone at 24 h. The killing activities of 4 mg/mL methylprednisolone on Nalm6, Pfeiffer, and U2932 were higher than that of 0.75 mg/ml group, while the killing activity of 12 mg/ml methylprednisolone was lower than that of 2.25 mg/ml dexamethasone at 48 h. However, the killing activities of different doses of methylprednisolone on EHEB tumor cells were lower than those of different doses of dexamethasone at 24 and 48 h. The average MFI and proportion of CD19 CAR-T cells under different concentrations of glucocorticoid the proliferation inhibition of CD19 CAR-T cells by dexamethasone was higher than that of methylprednisolone. The proliferation inhibition of CD19 CAR-T cells of the two glucocorticoids in high concentration groups were more obvious than that in low concentration groups. When CD19 CAR-T cells were co-cultured with different tumor cells, the proportion and average MFI of CD19 CAR-T cells showed that the proliferation inhibition of dexamethasone was higher than that of methylprednisolone. The proliferation inhibition of CD19 CAR-T cells of the two glucocorticoids in high concentration groups was more obvious than that in low concentration groups. Conclusion: Dexamethasone inhibits the cell proliferation of CD19 CAR-T cells more than methylprednisolone during the targeting of different tumor cell lines. The inhibition effect of dexamethasone on the proliferation and amplification of CD19 CAR-T cells was greater than that of methylprednisolone during the targeting of CD19 CAR-T cells to different tumor cell lines. Moreover, the inhibition effect of the high dose group was more obvious.

[Effects of Sidaxue, a Miao ethnomedicine recipe, on apoptosis and pyrolysis of human fibroblast-like synovial cells in rheumatoid arthritis].

OBJECTIVE: To investigate the effects of Sidaxue (SX), a recipe in Miao ethnomedicine, on apoptosis and pyrolysis of human fibroblast-like synovial cells in rheumatoid arthritis (RA-FLS). METHODS: The target proteins related with RA and those involved in cell apoptosis and pyroptosis were searched in different online databases, and Venny software was used to obtain apoptosis- and pyroptosis-related proteins in RA. RA-apoptosis-pyroptosis protein interaction (PPI) network was constructed to identity the key target proteins related with apoptosis and pyroptosis in RA. Autodock vina software was used to perform molecular docking to verify the binding ability of the main active ingredients in SX with the apoptosis- and pyroptosis-related proteins. In the cell experiment, MH7A cells were treated with 5 mg/L TGT (positive control) or 5, 10, 20, and 40 mg/L SX, and the changes in cell migration and invasion abilities and expressions of apoptosis- and pyroptosis-related proteins were examined using wound healing assay, Transwell assay, ELISA and Western blotting. RESULTS: We identified 9 RA-related apoptotic target proteins, 15 RA-related pyroptosis target proteins, and 4 overlapping target proteins related with RA, apoptosis and pyroptosis. The main active ingredients in SX had a high affinity with the target proteins including TNF-α, Fas, and Bax. In MH7A cells, SX treatment concentration-dependently increased the cell inhibition rate at 24, 48 and 72 h (P < 0.05), and significantly lowered the cell migration ability at 6, 12 and 24 h (P < 0.05); treatment with 20 and 40 mg/L SX for 24 h obviously suppressed MH7A cell invasion (P < 0.05). SX treatment (20 and 40 mg/L) and TGT treatment both significantly lowered the expression levels of TNF-α, IL-1ß, and IL-18 in the cells (P < 0.05). The Bax/Bcl-2 ratio and Fas and FasL expressions were increased significantly in cells treated with 20 and 40 mg/L SX (P < 0.05), and caspase-1 expression was decreased significantly in cells treated with 5 and 40 mg/L SX (P < 0.05). CONCLUSION: SX can induce apoptosis and pyroptosison in RA-FLSs possibly by down-regulating the expressions of TNF-α, IL-1ß and IL-18, up-regulating the expressions of Bax, Fas, and FasL, and inhibiting Bcl-2 and caspase-1 protein expressions.

Temperature Tides Across the Mid-Latitude Summer Turbopause Measured by a Sodium Lidar and MIGHTI/ICON.

Local full diurnal coverage of temperature variations across the turbopause (~90-115 km altitude) is achieved by combining the nocturnal observations of a Sodium (Na) Doppler lidar on the Utah State University (USU) campus (41.7°N, 248.2°E) and NASA Michelson interferometer for global high-resolution thermospheric imaging (MIGHTI)/Ionospheric connection explorer (ICON) daytime observations made in the same vicinity. In this study, utilizing this hybrid data set during summer 2020 between June 12th and July 15th, we retrieve the temperature signatures of diurnal and semidiurnal tides in this region. The tidal amplitudes of both components have similar vertical variation with increasing altitude: less than 5 K below ~98 km but increase considerably above, up to 19 K near 104 km. Both experience significant dissipation near turbopause altitudes, down to ~12 K up to 113 km for the diurnal tide and ~13 K for the semidiurnal tide near 110 km. In addition, while the semidiurnal tidal behavior is consistent with the theoretical predictions, the diurnal amplitude is considerably larger than what is expected in the turbopause region. The tidal phase profile shows a dominance of tidal components with a long vertical wavelength (longer than 40 km) for the semidiurnal tide. On the other hand, the diurnal tide demonstrates close to an evanescent wave behavior in the turbopause region, which is absent in the model results and Thermosphere ionosphere mesosphere energetics and dynamics (TIMED)/Sounding of the atmosphere using broadband radiometry (SABER) observations.

[Status quo regarding the recruit requirements of public health-related employers and training scheme for undergraduates majored in preventive medicine in colleges and universities].

Objective: To investigate public health-related professions' recruit requirements and understand the training scheme for preventive medicine-majored undergraduates in colleges and universities in China. Methods: An online questionnaire was adopted to collect data on the recruit requirements of public health-related professions. Information on the core courses, internship, and practical training for preventive medicine-majored undergraduates was collected through universities' official websites. Results: The employers had a high requirement for education and specific majors. Also, administration professions attached importance to comprehensive quality, while professional positions to professional skills. 47.2% (67/142) employers made Talent Introduction Programs for the next three years. 20.0% (3/15) universities offered all the core courses required by national standards. For internship and practical training, both clinical practice and professional practice's median durations were 16 and 11 weeks, respectively. Conclusion: Employers showed significant demand for public health graduates and having a high requirement for personal ability and comprehensive quality. The universities should improve the setting of core courses and strengthen students' practical ability and overall quality.

[Activity comparison of humanized CD19 CAR-T cells with murine CD19 CAR-T on Nalm-6 cells and xenograft tumor model].

Objective: To compare the activity difference of the high affinity humanized CD19 chimeric antigen receptor (CAR)-T cells and murine CD19 CAR-T cells. Methods: Peripheral venous blood T cells from 8 healthy volunteers were collected and infected with humanized and murine CD19 CAR lentivirus. Human and murine CD19 CAR-T cells were prepared and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The cytotoxicity of CD3(+) T cells, humanized and murine CD19 CAR-T cells to NALM-6 cells was detected by lactate dehydrogenase assay. Thirty BAL B/c nude mice transplanted with NALM-6 cells were randomly divided into 3 groups with 10 mice in each group and injected humanized CD19 CAR-T cells, mouse CD19 CAR-T cells and control CD3(+) T cell via tail vein, respectively. The proportion of NALM-6 cells in peripheral blood and the proportion of CD19 CAR-T cells in T cells from the vein of the inner canthus were detected by flow cytometry. The overall survival of BAL B/c nude mice was observed. Results: The proliferation of mouse and humanized CD19 CAR-T cells were (68.50±0.93)% and (80.63±1.41)%, respectively (t=20.353, P<0.001) after cultured in vitro for 24 hours, and were (91.38±1.41)% and (148.13±1.25)%, respectively (t=85.364, P<0.001) after cultured for 48 hours. When the effect to target ratio was 1∶1, there was no difference between the humanized and murine CD19 CAR-T cell group after co-culture for 24 hours (P=0.169), while the killing activity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells (P<0.01) after 48 hours of co-culture. When the effect to target ratio was 4∶1, the cytotoxicity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells in co-culture for 24 and 48 hours (P<0.01). On the seventh day of CD19 CAR-T cell therapy, the proportion of NALM-6 cells in the peripheral blood of BAL B/c nude mice decreased to the lowest level in the humanized CD19 CAR-T cell group and the murine CD19 CAR-T cell group. After 21 days, the proportion of NALM-6 cells in the murine CD19 CAR-T cell group was higher than that in the humanized CD19 CAR-T cell group (P(21 d)=0.001, P(28 d)<0.001, P(35 d)<0.001). The proportion of humanized and murine CD19 CAR-T cells in the peripheral blood reached the peaks after 7 days of therapy, and the proportion of humanized CD19 CAR-T cells was higher than that of murine CAR-T cells (P(7 d)=0.002). The CD19 CAR-T cells disappeared in the peripheral blood in the murine CD19 CAR-T cell group after 14 days of therapy, while in the humanized CD19 CAR-T cell group it disappeared after 21 days of therapy. The median survival of BAL B/c nude mice in the murine CD19 CAR-T cell group and the humanized CD19 CAR-T cell group was 42 days and 63 days, respectively (χ(2)=15.382, P<0.001). Conclusions: High affinity humanized CD19 CAR-T cells have stronger proliferation, higher cytotoxicity and longer survival time compared with those of murine CD19 CAR-T cells. The results indicate that the clinical efficacy of humanized CD19 CAR-T cells would be better than that of murine CD19 CAR-T cells.