Resumen Antecedentes: La enfermedad de injerto contra huésped (EICH) es una complicación importante después del trasplante alogénico de células madre hematopoyéticas. Objetivos: Para dilucidar el papel de TLR4, el principal receptor de LPS bacteriano, en el desarrollo de GVHD, construimos un modelo de GVHD en ratones knockout para TLR4 (TLR4-/-) y monitoreamos el quimerismo celular. Métodos: En este estudio, usamos PCR para identificar si se establecieron ratones knockout para TLR4 (TLR4-/-). Antes del trasplante, pretratamos a los ratones con irradiación para obtener la dosis de irradiación adecuada. Se aplicó citometría de flujo para medir el estado de quimerismo, las distribuciones de APC y células T en ratones receptores TLR4+/+ y TLR4-/-. Resultados: El estado general de los receptores de TLR4-/- fue mejor que el de los receptores de TLR4+/+, y los ratones receptores de TLR4-/- mostraron manifestaciones de GVHD menos graves que los ratones receptores de TLR4+/+. La mayoría de las APC y las células T en el bazo del ratón huésped se derivaron de las células del donante, y las células T CD4+, incluidas las células T de memoria, se encontraban en su mayoría en los ratones huéspedes. Conclusión: En general, nuestros datos muestran que la eliminación de TLR4 atenuó el desarrollo de GVHD, lo que sugiere que TLR4 podría usarse como un nuevo objetivo y paradigma terapéutico en las terapias de GVHD.

Abstract Background: Graft-versus-host disease (GVHD) is a major complication after allogeneic hematopoietic stem cell transplantation. Objective: To elucidate the role of Toll-like receptor 4 (TLR4), the major receptor for bacterial lipopolysaccharide, in the development of GVHD, we constructed a GVHD model in TLR4 knockout (TLR4-/-) mice and monitored the cell chimerism. Methods: In this study, we used polymerase chain reaction to identify whether TLR4 knockout (TLR4-/-) mice were established. Before transplantation, we pretreated mice with irradiation so as to obtain an appropriate irradiation dose. Flow cytometry was applied to measure the chimerism status, the distributions of antigen-presenting cells (APCs), and T-cells in TLR4+/+ and TLR4-/- recipient mice. Results: The general condition of TLR4-/- recipients was better than that of TLR4+/+ recipients, and the TLR4-/- recipient mice showed less severe GVHD manifestations than the TLR4+/+ recipient mice. Most of the APCs and T-cells in the host mouse spleen were derived from donor cells, and CD4+ T-cells, including memory T-cells, were in the majority in host mice. Conclusion: In general, our data show that TLR4 deletion attenuated GVHD development, which suggests that TLR4 could be used as a novel target and therapeutic paradigm in GVHD therapies.

Abstract Objectives: The degree of endolymphatic hydrops in Meniere’s disease may be related to hearing loss. However, the results of prior studies have been inconsistent. We aimed to investigate the relationship between endolymphatic hydrops and hearing loss characteristics in Meniere’s disease. Methods: This study included 54 patients (62 ears) with Meniere’s disease. Patients underwent three-dimensional real inversion recovery sequences for magnetic resonance imaging and puretone audiometry. Endolymphatic hydrops were assessed according to Gurkov’s criteria (2011). Correlations between different degrees of endolymphatic hydrops and pure-tone audiometry, as well as staging, were analysed. Results: Pure tone audiometry and staging were higher for vestibular endolymphatic hydrops complicated by cochlear ones than isolated cochlear or vestibular hydrops (both p<0.05). There was no significant correlation between vestibular endolymphatic hydrops and pure-tonal audiometry or staging (all p > 0.05). The degree of hydrops in the middle turn was correlated with the mid-frequency hearing threshold (p<0.05). The degree of cochlear hydrops was correlated with the audiometry, low-frequency hearing thresholds, mid-frequency hearing thresholds and staging (p<0.01). Conclusion: The types and sites of endolymphatic hydrops based on a 3D-real inversion recovery sequence can be used to indicate the degree of hearing loss in patients with Meniere's disease. Level of evidence: Level III.

Abstract Background The main features of polycystic ovary syndrome (PCOS) are abnormal follicular development and ovulatory dysfunction, which are caused by excessive apoptosis of ovarian granulosa cells. Acupuncture has been shown to improve follicular development abnormalities in patients with PCOS, but its mechanism is unknown. This study hypothesized that the mechanism of acupuncture on follicular development abnormalities in PCOS patients is the inhibition of granulosa cell apoptosis through LncMEG3-mediated regulation of miR-21-3p. Methods A PCOS-like rat model was established using subcutaneous injection of dehydroepiandrosterone (DHEA). Acupuncture was performed on rats for 15 d (CV-4, RN-3, CV-6, SP-6 and EX-CA 1). Ovarian morphology was observed by HE staining, and sex hormone and AMH levels were detected by ELISA. Primary granulosa cells were isolated from each group of rats to assess the association of acupuncture treatment, LncMEG3, miR-21-3p, and granulosa cell apoptosis in rats with PCOS. Results LncMEG3 and miR-21-3p were highly expressed in the ovarian granulosa cells of rats with PCOS, and LncMEG3-mediated regulation of miR-21-3p was involved in the development of PCOS in rats. Silencing of MEG3 attenuated sex hormone dysregulation and ovarian histopathological changes in PCOS rats and promoted follicle cell development and maturation. In addition, silencing MEG3 increased the viability and number of granulosa cells. In addition, silencing MEG3 further inhibited early and late apoptosis of ovarian granulosa cells in PCOS rats. Acupuncture improved polycystic ovarian morphology and sex hormone levels in PCOS rats. Acupuncture intervention increased the viability and number of granulosa cells. Acupuncture intervention inhibited early and late apoptosis of ovarian granulosa cells in PCOS rats by targeting miR-21-3p via LncMEG3. Conclusion These results suggest that acupuncture can downregulate LncMEG3, thereby targeting and regulating miR-21-3p to suppress early and late granulosa cell apoptosis and normalize their proliferation. These factors ultimately compensate for abnormal follicular development. These findings shed light on the clinical potential of acupuncture as a safe treatment for follicular developmental abnormalities in PCOS. Highlights LncMEG3-mediated inhibition of miR-21-3p regulates ovarian granulosa cell apoptosis. LncMEG3 and miR-21-3p are involved in the occurrence and development of PCOS-related abnormal follicular development. CuONPs induce co-occurrence of autophagy activation and autophagic flux blockade. Acupuncture can improve the sex hormone levels and follicular development in the context of PCOS. The underlying mechanism of acupuncture in the treatment of PCOS abnormal follicular development was revealed.

Abstract The image of an athlete's ankle joint injury can help to check whether the athlete's ankle joint is damaged, and plays a very important role in clinical diagnosis. To address the problem of poor segmentation effect of traditional athletes' ankle injury image segmentation algorithm, an ankle injury image segmentation algorithm based on residual double attention U-Net model is proposed. First, the region of interest is extracted from the original ankle injury image. After translation, rotation and turnover, the image data is expanded. Second, the residual structure is used to adjust the gradient propagation and residual feedback of the segmentation framework, extract the attribute information in the region of interest, and combine the two to retain more image features. Finally, combined with the double attention module to improve the weight ratio of image features, the athlete ankle injury image segmentation is realized in the image segmentation framework based on residual double attention U-Net model. The results demonstrate that the maximum values of DSC, ASSD, PM, and CR for the proposed algorithm are 0.93, 0.1, 0.96, and 0.95, respectively, and the F1 score is 95.7%, indicating that the segmentation effect of this algorithm is closer to the theoretical segmentation effect, and higher precision in segmentation, and the segmented image has a high degree of similarity to the original image, resulting in excellent segmentation performance.

Abstract Background Currently, only a few studies have described the general characteristics of patients with primary Sjögren's syndrome (pSS) who tested negatives for anti-SSA and anti-SSB antibodies. We aimed to further investigate the clinical characteristics of these patients in a large sample. Methods Data from patients with pSS who were treated at a tertiary hospital in China between 2013 and 2022 were retrospectively analyzed. Clinical characteristics of the patients were compared between those with and without anti-SSA and anti-SSB antibody negativity. Factors associated with anti-SSA and anti-SSB negativity were identified by logistic regression analysis. Results Overall, 934 patients with pSS were included in this study, among whom 299 (32.0%) tested negative for anti-SSA and anti-SSB antibodies. Compared with patients testing positive for anti-SSA or anti-SSB antibodies, that testing negative for the two antibodies had a lower proportion of females (75.3% vs. 90.6%, p < 0.001) and thrombocytopenia (6.7% vs. 13.6%, p = 0.002), but a higher proportion of abnormal Schirmer I tests (96.0% vs. 89.1%, p = 0.001) and interstitial lung disease (ILD) (59.2% vs. 28.8%, p = 0.001). Anti-SSA and anti-SSB negativity was positively associated with male sex (odds ratio [OR] = 1.86, 95% confidence interval [CI]: 1.05, 3.31), abnormal Schirmer I tests (OR = 2.85, 95% CI: 1.24, 6.53), and ILD (OR = 2.54, 95% CI: 1.67, 3.85). However, it was negatively related to thrombocytopenia (OR = 0.47, 95% CI: 0.24, 0.95). Conclusion Approximately one third of pSS patients had anti-SSA and anti-SSB negativity. pSS patients testing negative for anti-SSA and anti-SSB showed a higher risk of abnormal Schirmer I tests and ILD, but a lower risk of thrombocytopenia.

Resumo O objetivo do estudo foi investigar os efeitos da atividade física (AF) na variabilidade da frequência cardíaca (VFC) em crianças e adolescentes. Realizamos uma pesquisa nas bases Web of Science, PubMed, ScienceDirect, Springer-Link e EBSCO-host. A Escala Newcastle-Ottawa revisada foi utilizada para avaliar o risco de enviesamento. Um total de 21 estudos foi incluído. De forma geral, foram encontradas associações de médio porte entre AF e baixa frequência e alta frequência em crianças e adolescentes. O alto nível de AF teve um desvio padrão significativamente maior dos intervalos e raiz da média da soma dos quadrados de diferenças entre os intervalos RR adjacentes em crianças e adolescentes. Os efeitos de AF sobre VFC foram consistentes em crianças e adolescentes. Nossa revisão sistemática e meta-análise revelou que AF e VFC em crianças e adolescentes são de médio porte. Promover a participação de crianças e adolescentes em atividade física de moderada à vigorosa (AFMV) aumentará a atividade nervosa parassimpática e diminuirá a atividade nervosa simpática. Nossas descobertas apoiam a motivação de crianças e adolescentes a se envolverem mais na AFMV em suas vidas diárias para melhorar o funcionamento do sistema nervoso autônomo e promover a segurança cardiovascular.

Abstract The aim of the study was to investigate the effects of physical activity (PA) on heart rate variability (HRV) in children and adolescents. We conducted a research of Web of Science, PubMed, ScienceDirect, Springer-Link and EBSCO-host. The revised Newcastle-Ottawa Scale was used in an investigative analysis to assess bias risk. A total of 21 studies were included. Overall, medium-sized associations were found between PA and low frequency and high frequency in children and adolescents. High PA level had significantly higher standard deviation of RR intervals and root of the mean of the sum of the squares of differences between adjacent RR intervals in children and adolescents. The effects of PA on HRV were consistent in children and adolescents. Our systematic review and meta-analysis revealed medium-sized between PA and HRV in children and adolescents. Promoting children’s and adolescents’ participation in moderate-to-vigorous physical activity (MVPA) will increase parasympathetic nerve activity and decreased sympathetic nerve activity. Our findings support motivating children and adolescents to engage in more MVPA in their daily lives to improve autonomic nervous system function and promote cardiovascular safety.

Resumo O objetivo do estudo foi investigar os efeitos da atividade física (AF) na variabilidade da frequência cardíaca (VFC) em crianças e adolescentes. Realizamos uma pesquisa nas bases Web of Science, PubMed, ScienceDirect, Springer-Link e EBSCO-host. A Escala Newcastle-Ottawa revisada foi utilizada para avaliar o risco de enviesamento. Um total de 21 estudos foi incluído. De forma geral, foram encontradas associações de médio porte entre AF e baixa frequência e alta frequência em crianças e adolescentes. O alto nível de AF teve um desvio padrão significativamente maior dos intervalos e raiz da média da soma dos quadrados de diferenças entre os intervalos RR adjacentes em crianças e adolescentes. Os efeitos de AF sobre VFC foram consistentes em crianças e adolescentes. Nossa revisão sistemática e meta-análise revelou que AF e VFC em crianças e adolescentes são de médio porte. Promover a participação de crianças e adolescentes em atividade física de moderada à vigorosa (AFMV) aumentará a atividade nervosa parassimpática e diminuirá a atividade nervosa simpática. Nossas descobertas apoiam a motivação de crianças e adolescentes a se envolverem mais na AFMV em suas vidas diárias para melhorar o funcionamento do sistema nervoso autônomo e promover a segurança cardiovascular.

Abstract The aim of the study was to investigate the effects of physical activity (PA) on heart rate variability (HRV) in children and adolescents. We conducted a research of Web of Science, PubMed, ScienceDirect, Springer-Link and EBSCO-host. The revised Newcastle-Ottawa Scale was used in an investigative analysis to assess bias risk. A total of 21 studies were included. Overall, medium-sized associations were found between PA and low frequency and high frequency in children and adolescents. High PA level had significantly higher standard deviation of RR intervals and root of the mean of the sum of the squares of differences between adjacent RR intervals in children and adolescents. The effects of PA on HRV were consistent in children and adolescents. Our systematic review and meta-analysis revealed medium-sized between PA and HRV in children and adolescents. Promoting children’s and adolescents’ participation in moderate-to-vigorous physical activity (MVPA) will increase parasympathetic nerve activity and decreased sympathetic nerve activity. Our findings support motivating children and adolescents to engage in more MVPA in their daily lives to improve autonomic nervous system function and promote cardiovascular safety.

Abstract Lactic acid bacteria (LAB) can produce extracellular polysaccharides that can be used as thickeners, emulsifiers, and gels in foods. The immobilization technology can effectively simplify and improve the separation and regeneration of extracellular polysaccharides. In this study, six strains of LAB, including the extracellular polysaccharide-producing strain Lactobacillus plantarum, were screened. The different immobilized carriers of sodium alginate, agar, gelatin-glutaraldehyde, and carrageenan were assessed for fermentation by these six LAB. Different diameters of beads were selected, and the optimum culture temperature, carrier concentration, and inoculum volume were evaluated. The results showed that the agar-embedding method was the best immobilization method and the optimum process conditions were as follows: 3% inoculum of L. plantarum, 30 g/L of agar as immobilization carrier with a particle diameter of 1 cm, and fermentation at 37 °C for 24 h. Through orthogonal experiments, the maximum exopolysaccharide (EPS) yield obtained was 1489.9 mg/L.

Abstract Background: The internal NAD(P)H dehydrogenase (NDA) gene family was a member of the NAD(P)H dehydrogenase (ND) gene family, mainly involved in the non-phosphorylated respiratory pathways in mitochondria and played crucial roles in response to abiotic stress. Methods: The whole genome identification, structure analysis and expression pattern of NDA gene family were conducted to analyze the NDA gene family. Results: There were 51, 52, 26, and 24 NDA genes identified in G. hirsutum, G. barbadense, G. arboreum and G. raimondii, respectively. According to the structural characteristics of genes and traits of phylogenetic tree, we divided the NDA gene family into 8 clades. Gene structure analysis showed that the NDA gene family was relatively conservative. The four Gossypium species had good collinearity, and segmental duplication played an important role in the evolution of the NDA gene family. Analysis of cis-elements showed that most GhNDA genes contained cis-elements related to light response and plant hormones (ABA, MeJA and GA). The analysis of the expression patterns of GhNDA genes under different alkaline stress showed that GhNDA genes were actively involved in the response to alkaline stress, possibly through different molecular mechanisms. By analyzing the existing RNA-Seq data after alkaline stress, it was found that an NDA family gene GhNDA32 was expressed, and then theGhNDA32 was silenced by virus-induced gene silencing (VIGS). By observing the phenotype, we found that the wilting degree of silenced plants was much higher than that of the control plant after alkaline treatment, suggesting that GhNDA32 gene was involved in the response to alkaline stress. Conclusions: In this study, GhNDAs participated in response to alkaline stress, especially NaHCO3 stress. It was of great significance for the future research on the molecular mechanism of NDA gene family in responding to abiotic stresses.

Abstract Aim The purpose of this study was to evaluate DHA’s effect and relative mechanisms in diabetic keratopathy (DK) treatment by vitro study. Methods and materials: Using high glucose to make DK cell model; measuring cell apoptosis by flow cytometry and Hoechst 33342 staining; evaluating cell migration abilities by wound healing assay; relative gene and proteins including PEDF, VEGF, Caspase-3, Caspase-9 and NF-κB(p65) expression by RT-qPCR and WB assay; And evaluatign PDEF and p-NF-κB(p65) protein expression by immunofluorescence detection. Results: After high glucose treatment, TKE-2 cell apoptosis was significantly increased and cell migration ability was significantly depressed with PEDF, Caspase-3, Caspase-9 and NF-κB(p65) gene and protein significantly increasing and VEGF expression significantly depressing (P < 0.001, respectively); with DHA supplement, TKE-2 cell biological activities were significantly improved with relative gene and proteins significantly changed; However, with DHA and PEDF supplement, DHA’s treatment was disappeared with PEDF, Caspase-3, Caspase-9 and NF-κB(p65) gene and protein significantly increasing and VEGF expression significantly depressing (P < 0.001, respectively). Conclusion: DHA had effects to improved DK by regulation PEDF pathway in vitro study.

Abstract Character strengths could effectively prevent negative psychological outcomes in adults. However, there was little research conducted among early adolescents. The present study aimed to explore character strengths that were independently related to fewer behavior problems in early adolescents. In total, 521 early adolescents (mean age 10.92 ± 0.04, range 10–12 years) were recruited from primary schools in Sichuan, China. Character strengths were measured using the Values in Action Inventory of Strengths for Youth (VIA-Youth). Behavior problems were measured using the Conners Parent Symptom Questionnaire (PSQ). The results showed that behavior problems were negatively correlated with character strengths ( r = −0.14 to −0.3, p < 0.05 Bonferroni corrected). Character strengths explained a significant proportion of additional variance (14–22%) in five types of behavior problems after controlling the effect of demographic factors (residence, left-behind experiences, maternal education level). Moreover, several specific character strengths showed an independent contribution ( β = −0.34 to −0.14 for self-regulation, perseverance, zest, humility, and leadership; β = 0.21 to 0.34, for hope; all p < 0.05) to behavior problems. Our study revealed that character strengths were protective factors against behavior problems in early adolescents.

Abstract Megalobrama Amblycephala is an important fishery resource. The processing of Megalobrama Amblycephala often generates many underutilized leftovers including bone which contents abundant active calcium. This study investigated the optimization for extraction conditions of active calcium from Megalobrama Amblycephala bone. The active calcium was extracted from Megalobrama Amblycephala bone using citric acid and malic acid. The response surface methodology was applied to optimize the extraction technology parameters including extraction temperature, extraction time and ratio of citric acid and malic acid. Results showed that, the optimal conditions of extracting active calcium from Megalobrama Amblycephala bone were as follows: extraction temperature, 113 °C, extraction time, 0.8 h, ratio of citric acid and malic acid, 2: 1. Under these conditions, the yield of active calcium was 90.11%. This study has provided a reference for efficiently obtaining of active calcium and utilization of Megalobrama Amblycephala bone resource.

Abstract Apigenin is one kinds of natural flavonoids from plants. Many researches showed this compound has potential antioxidant, anti-inflammatory and anticancer functions. This study aimed to investigate the effects of apigenin on proliferation, migration, invasion, epithelial mesenchymal transition (EMT) in glioma cells, and to reveal the underlying mechanism of apigenin. MTT assay indicated the proliferation rate of glioma cells U251 was significantly decreased with the treatment of apigenin in a dose-dependent manner. In addition, apigenin could inhibit the cell invasion, wound healing and decrease the expression of EMT-related proteins, NEDD9, p-FAK and p-AKT. Quantitative real-time PCR showed miR-103a-3p was remarkably increased in the presence of apigenin. Inhibition of miR-103a-3p attenuated the effects of apigenin on U251 cells. Dual-luciferase assay and western blot further demonstrated that NEDD9 was a direct target of miR-103a-3p. Moreover, knockdown NEDD9 almost reversed the activities of silenced miR-103a-3p in transwell invasion, wound healing and regulation of NEDD9/FAK/AKT axis. In conclusion, apigenin could inhibit glioma cell proliferation, migration, invasion and EMT via activating of miR-103a-3p and targeting NEDD9/FAK/AKT pathway.

ABSTRACT Purpose: This study investigated the effects of oral administration of Clostridium butyricum (C. butyricum) on inflammation, oxidative stress, and gut flora in rats with hepatic ischemia reperfusion injury (HIRI). Methods: The rats from C. butyricum group were given C. butyricum for 5 days. Then, hepatic ischemia for 30 min and reperfusion for 6 h were performed in all the rats. After the animals were sacrificed, alanine transaminase (ALT), aspartate aminotransferase (AST), lipopolysaccharide (LPS) in serum, short-chain fatty acids (SCFAs), and gut microbiota composition in feces, and malondialdehyde (MDA), glutathione (GSH), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), Toll-like receptor 4 (TLR4), nuclear factor-kappa Bp65 (NF-κBp65) and histological analysis in the liver were performed. Results: The rats given C. butyricum showed decreased ALT, AST, LPS, and MDA; improved GSH and histological damage; changes in SCFAs; declined TNF-α, IL-6, TLR4, and pNF-κBp65/NF-κBp65; and changes in the gut microbial composition, which decreased the Firmicutes/Bacteroidetes ratio and increased the relative abundance (RA) of probiotics. Conclusions: C. butyricum supplementation protected against HIRI by regulating gut microbial composition, which contributed to the decreased LPS and attenuation of inflammation and oxidative stress. These indicate C. butyricum may be a potent clinical preoperative dietary supplement for HIRI.

Abstract Objectives: Depression is common after both lacunar stroke and non-lacunar stroke and might be associated with lesion locations as proven by some studies. This study aimed to identify whether lesion location was critical for depression after both lacunar and non-lacunar strokes. Methods: A cohort of ischemic stroke patients was assigned to either a lacunar stroke group or a non-lacunar stroke group after a brain MRI scan. Neurological deficits and treatment response was evaluated during hospitalization. The occurrence of depression was evaluated 3 months later. Logistic regressions were used to identify the independent risk factors for depression after lacunar and non-lacunar stroke respectively. Results: 83 of 246 patients with lacunar stroke and 71 of 185 patients with non-lacunar stroke developed depression. Infarctions in the frontal cortex, severe neurological deficits, and a high degree of handicap were identified as the independent risk factors for depression after non-lacunar stroke, while lesion location was not associated with depression after lacunar stroke. Conclusion: The main determinants for depression after lacunar and non-lacunar stroke were different. Lesion location was critical only for depression after non-lacunar stroke.