ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

Metastasis is the main risk factor for poor prognosis of laryngeal squamous cell carcinoma (LSCC) .Chemokines are closely related to metastasis in the tumor microenvironment.CXCL8 is a cyto-kine-like secreted protein that plays key roles in the malignant development of a variety of tumors,but has not been elucidated in LSCC.In this paper,we elucidated the role of CXCL8 in LSCC cells and found miRNAs that targeted CXCL8,which may become new targets for the diagnosis and treatment of LSCC.Firstly,the GEPIA showed that CXCL8 was highly expressed in head and neck cancer (P<0.05).The real-time fluorescence quantification (qRT-PCR) found that CXCL8 was highly expressed in LSCC cells.The enzyme-linked immunoassay also found that CXCL8 was highly secreted in the superna-tant of LSCC cells (P<0.001) .Then,the CCK8 assay confirmed that knockdown of CXCL8 significant-ly inhibited the proliferation of FD-LSC-1 and AMC-HN8 cells (the average inhibition rate was 34.0％and 19.5％,respectively);The EdU assay also confirmed that knockdown of CXCL8 significantly inhibi-ted the proliferation of LSCC cells (P<0.05) .The transwell assay confirmed that knockdown of CXCL8 also significantly inhibited the migration and invasion of FD-LSC-1 cell (average inhibition rate was 40.0％,38.5％,respectively);Knockdown of CXCL8 also significantly inhibited the migration and inva-sion of AMC-HN8 cell (average inhibition rate was 37.5％,53.5％,respectively ) .The analysis of bioinformatics predicted that CXCL8 may be a target of miR-513b-5p.The dual luciferase reporter assay confirmed that miR-513b-5p could bind to the CXCL8-3'UTR.QRT-PCR assay also confirmed that over-expression of miR-513b-5p could decrease the 60％ of the CXCL8 expression (P<0.01) .Cell function rescue assays found that overexpressed of CXCL8 could effectively reversed proliferation,migration and invasion of LSCC cells weakened by miR-513b-5p (P<0.05) .In summary,miR-513b-5p inhibited the proliferation,migration and invasion of LSCC cells by targeting CXCL8.

Objective:To understand the prevalence of insufficient physical activity among adults aged ≥18 years in China and to explore the influencing factors.Methods:The China Chronic Disease and Risk Factor Surveillance was conducted in 298 counties/districts in China in 2018, covering 31 provinces (autonomous regions, municipalities). A multi-stage stratified cluster random sampling method was used to select permanent residents aged ≥18 years. A questionnaire including Global Physical Activity Questionnaire was used to collect information about the participants' demographic characteristics and physical activity through face-to-face interview. A total of 183 769 participants completed the survey. After complex data weighting, the prevalence of insufficient physical activity, occupation, transportation, and leisure-time physical activity time was analyzed. Multivariate logistic regression models were used to analyze the influencing factors related to insufficient physical activity.Results:The prevalence of insufficient physical activity among adults aged ≥18 years was 22.3% (95% CI: 20.9%-23.7%) in China in 2018, with males [24.4% (95% CI: 23.0%-25.8%)] significantly higher than females [20.2% (95% CI: 18.6%-21.8%)]. Adults aged 70 years and above [28.4% (95% CI: 26.9%-29.9%)] were significantly higher than adults in other age groups, followed by adults aged 18-29 years [26.4% (95% CI: 24.4%- 28.3%)] and 30-39 years [23.4% (95% CI: 21.5%-25.3%)], and tended to increase with increasing education and total sedentary behavior time ( P<0.001). The weekly occupation, transportation, and leisure-time physical activity time appeared 958.6 (95% CI: 911.4-1 005.8) minutes, 234.5 (95% CI: 224.7- 244.2) minutes, and 88.6 (95% CI: 83.5-93.7) minutes, respectively. Multivariate logistic regression analysis showed that males, adults living in rural areas or northern China, ≥70 years, with junior high school education, an annual household income per capita <6 000 yuan and institutional/clerical/ technical occupation and longer total sedentary behavior time were related to a higher prevalence of insufficient physical activity. Conclusions:In China, over one-fifth of the adults had lower physical activity levels. Adults who are male, young adults, more educated, institutional/clerical/technical occupation, and with more extended total sedentary behavior are the populations that need to be focused on to promote physical activity-related health.

Currently, the gut-organ axis has become a hot research topic. As increasing attention has been paid to the role of gut microbiota in the health of organs, the complex and integrated dialogue mechanism between the gastrointestinal tract and the associated microbiota has been demonstrated in more and more studies. Skin as the largest organ in the human body serves as the primary barrier protecting the human body from damage. The proposal of the gut-skin axis has established a bidirectional link between the gut and the skin. The disturbance of gut microbiota can lead to the occurrence of skin diseases, the mechanism of which is complex and may involve multiple pathways in immunity, metabolism, and internal secretion. According to the theory of traditional Chinese medicine(TCM), the connection between the intestine and the skin can be established through the lung, and the interior disorders will definitely cause symptoms on the exterior. This paper reviews the research progress in the gut-skin axis and its correlation with TCM theory and provides ideas and a basis for cli-nical treatment and drug development of skin and intestinal diseases.
Humans ; Medicine, Chinese Traditional ; Gastrointestinal Tract ; Skin Diseases/drug therapy* ; Gastrointestinal Microbiome

Objective:To construct and validate a prediction model for the risk of intermediate cesarean delivery for primiparous women with failed vaginal trial of labor.Methods:Clinical data of 6 128 pregnant women who gave birth in the Affiliated Hospital of Jining Medical College from January 2019 to December 2020 were collected. The puerpera was randomly divided into train set ( n=4 290) and validation set ( n=1 838). The factors influencing the conversion to cesarean section in primiparous women with failed vaginal trial of labor were analyzed with univariate and binary multivariate logistic regression, and a risk prediction model was established based on the influencing factors. The predictive power of the model was assessed by receiver operating characteristic (ROC) curve and the Hosmer-Lemeshow goodness-of-fit test in train set and validation set. Results:Among 6 128 pregnant women 1 042 cases failed in vaginal trial of labor and were transferred to cesarean section. Univariate analysis showed age, occupation, gestational weight gain, days of gestation, body temperature before delivery, fetal heart condition at delivery, fetal abdominal circumference, Bishop score, premature rupture of membranes, gestational illness, mode of induction of labor, labor analgesia, and fetal orientation were significantly associated with converting to cesarean delivery (all P<0.05). The multivariate binary logistic regression analysis showed that the age, gestational weight gain, body temperature, gestational co-morbidities, days of gestation, premature rupture of membranes, amniotic fluid contamination, induction of labor, and abnormal occipital position were independent risk factors for intermediate cesarean delivery ( OR=1.03-8.06, all P<0.05); while height, occupation, Bishop score, and labor analgesia were protective factors for intermediate cesarean delivery ( OR=0.17-0.96, all P<0.05). A risk prediction model was constructed based on the risk factors and protective factors. In train set, the area under the ROC curve(AUC) of the model was 0.902 (95% CI: 0.89-0.92, P<0.001), with the best cutoff value of 0.138, the sensitivity and specificity were 0.837 and 0.825, respectively; and the Hosmer-Lemeshow goodness-of-fit test showed P=0.192. In validation set the AUC of the model was 0.917 (95% CI: 0.90-0.93, P<0.001), and the sensitivity and specificity were 0.826 and 0.851, respectively; the total correct rate of the model was 87.21% (1 603/1 838). Conclusion:The risk prediction model of failed vaginal trial of labor in primiparous women for intermediate cesarean delivery constructed in this study has good clinical prediction efficacy and high correctness rate.

Objective:To find the preferences of nursing staff when they provide " Internet + nursing service" for elderly patients.Methods:By means of the discrete choice experiment, a questionnaire was designed to investigate the preferences of nursing staff when they provide " Internet + nursing services" for elderly patients. In-service nursing staff from 8 medical and health institutions in Ningbo city and Wenzhou city were selected by random sampling, for an online questionnaire survey conducted from May to June 2022. The preferences of nursing staff on their service income, service content, service distance, service continuity and residence status of elderly patients when providing " Internet + nursing service" were analyzed by mixed logit regression.Results:A total of 420 valid questionnaires were collected. Compared to 50 yuan/order, nursing staff preferred to a price of 150 yuan/order ( β= 1.22, P<0.001) nursing services; Compared to specialized nursing services, nursing staff preferred to the routine care ( β= 0.86, P<0.001) and health promotion ( β= 0.86, P<0.05) service; Compared to<5 km, nursing staff were unwilling to provide nursing services for elderly patients at distances of 5-10 km and 11-15 km ( β=-0.66, P<0.05; β=-0.95, P<0.001) ; Compared to 1-2 visits per month, nursing staff preferred not to provide continuing care services ( β=-0.70, P<0.05); Compared to homestay with the patient family, nursing staff preferred to provide nursing services for elderly patients residing in nursing homes ( β= 1.21, P<0.001) . Conclusions:Considering the preference of nursing staff tend to provide " Internet + nursing service" for the elderly patients with services featuring appropriate price, non-specialist care, close distance, low continuity (moderate intensity used as the reference) and security assurance for practice.

AIM: To optimize the technique of intravenous injection of Evans blue and retinal preparations in mice, improving the accuracy and repeatability of staining experiment of retinal preparations.METHODS: C57BL/6 male mice were intravenous injected with 10g/L(1%)Evans Blue 0.3mL and circulated in vivo for 10 or 20min, and the eyes were removed after sacrificed and fixed in 4% paraformaldehyde for 20, 40 or 60min. When failure of intravenous injection, the experiment was remediated by intraperitoneal injection of 1% Evans Blue 0.3mL, circulated in vivo for 3h and fixed for 60min to observe morphology, distribution and leakage of the retinal vessels. Besides, we compared the morphology, distribution and leakage of the retinal vessels after intravenous injection with those after intraperitoneal injection to determine the optimal conditions for in vivo circulation time and retinal preparations.RESULTS: After intravenous injection, compared to the retinal vascular condition under 20min in vivo circulation time of Evans blue and 20 or 40min of fixation, with 10min of in vivo Evans blue circulation and 60min of fixation, the morphology of retinal vascular was more intact with less retinal vascular leakage, and the vascular branches are clear. When intravenous injection failed, remediated results from intraperitoneal injection showed that the morphology and distribution of retinal vessels were intact. There was no significant difference in morphology, distribution and leakage of the retinal vessels after 3h of intraperitoneal Evans blue circulation compared to 10min intravenous Evans blue circulation.CONCLUSION: This experiment optimizes the protocol, improves the accuracy and reproducibility of retinal preparations, and provides a reference for the study of related retinal vascular diseases.

Animals ; Rats ; Explosions ; Proteomics ; Autophagy

Objective To investigate the relationship between changes in protein tyrosine kinase 7 (PTK7) and receptor tyrosine kinase-like orphan receptor 2 ( ROR2) expression in the brainstem and the pathogenesis of amyotrophic lateral sclerosis (ALS). Methods Forty-four human superoxide dismutase 1( hSODl)-G93A mutant ALS transgenic mice were selected, and an equal number of wild-type littermates was used as control. The brainstems were isolated at da)' 70, day 95, day 108 and da)' 122 after birth, and the morphology of frypoglossal nucleus (12N) and nucleus of facial nerve(7N) neurons in the brainstem of the model mice were observed by Nissl staining. The mRNA and protein expression of PTK7 and ROR2 were detected by RT-PCR and Western blotting respectively, and the cellular localization and distribution of PTK7 and ROR2 in 12N and 7N were observed by immunofluorescence double-labeling technique. Results The result of Nissl staining showed that Nissl bodies in the neurons reduced distinctly with vacuolar degeneration of neurons, cell body atrophy and nuclear volume reduction in the 12N and 7N brainstems of ALS transgenic mice. RT-PCR result indicated that ROR2 and PTK7 mRNA level in the brainstem of ALS transgenic mice were up-regulated at da)' 70, da)' 95, day 108 and day 122 compared with wild-type littermates. Western blotting result showed that PTK7 protein was up-regulated at day 70, day 95, day 108 and day 122, ROR2 protein was up-regulated at day 70, day 95, day 108, and down-regulated at day 122 in the brainstem of ALS transgenic mice compared with wild-type littermates. Immunofluorescence result showed that ROR2/neuronal nuclei (NeuN)and PTK7/NeuN double positive cells, ROR2/glial fibrillary acidic protein (GFAP) and PTK7/GFAP double positive cells were observed in the 12N and 7N of the brainstem of ALS transgenic mice and wild-type mice, suggesting that ROR2 and PTK7 were expressed both in neurons and astrocytes. Conclusion PTK7 and ROR2 are abnormally expressed in the brainstem of ALS transgenic mice, which is closely related to the pathogenesis of ALS.