Abstract Background Spermatogonial stem cells (SSCs) are critical for sustaining spermatogenesis. Even though several regulators of SSC have been identified in rodents, the regulatory mechanism of SSC in humans has yet to be discovered. Methods To explore the regulatory mechanisms of human SSCs, we analyzed publicly available human testicular single-cell sequencing data and found that Ankyrin repeat and SOCS box protein 9 (ASB9) is highly expressed in SSCs. We examined the expression localization of ASB9 using immunohistochemistry and overexpressed ASB9 in human SSC lines to explore its role in SSC proliferation and apoptosis. Meanwhile, we used immunoprecipitation to find the target protein of ASB9 and verified its functions. In addition, we examined the changes in the distribution of ASB9 in non-obstructive azoospermia (NOA) patients using Western blot and immunofluorescence. Results The results of uniform manifold approximation and projection (UMAP) clustering and pseudotime analysis showed that ASB9 was highly expressed in SSCs, and its expression gradually increased during development. The immunohistochemical and dual-color immunofluorescence results displayed that ASB9 was mainly expressed in nonproliferating SSCs. Overexpression of ASB9 in the SSC line revealed significant inhibition of cell proliferation and increased apoptosis. We predicted the target proteins of ASB9 and verified that hypoxia-inducible factor 1-alpha inhibitor (HIF1AN), but not creatine kinase B-type (CKB), has a direct interaction with ASB9 in human SSC line using protein immunoprecipitation experiments. Subsequently, we re-expressed HIF1AN in ASB9 overexpressing cells and found that HIF1AN reversed the proliferative and apoptotic changes induced by ASB9 overexpression. In addition, we found that ABS9 was significantly downregulated in some NOA patients, implying a correlation between ASB9 dysregulation and impaired spermatogenesis. Conclusion ASB9 is predominantly expressed in human SSCs, it affects the proliferation and apoptotic process of the SSC line through HIF1AN, and its abnormal expression may be associated with NOA.

Abstract Under the influence of the COVID-19, people's awareness of physical health and immunity has increased significantly. Chitooligosaccharide is an oligomer of β-(1, 4)-linked D -glucosamine, furthermore, is one of the most widely studied immunomodulators. Chitooligosaccharide can be prepared from the chitin or chitosan polymers through enzymatically, chemically or physically processes. Chitooligosaccharide and its derivatives have been proven to have a wide range of biological activities including intestinal flora regulation, immunostimulant, anti-tumor, anti-obesity and anti-oxidation effects. This review summarizes the latest research of the preparation methods, biological activities in immunity and safety profiles of Chitooligosaccharide and its derivatives. We recapped the effect mechanisms of Chitooligosaccharide basing on overall immunity. Comparing the effects of Chitooligosaccharide with different molecular weights and degree of aggregation, a reference range for usage has been provided. This may provide a support for the application of Chitooligosaccharide in immune supplements and food. In addition, future research directions are also discussed.

Abstract Objective Abnormal complement activation is associated with periodontitis. W54011 is a novel non-peptide C5aR antagonist (C5aRA) that exhibits favorable anti-inflammatory effects in various inflammatory models. However, whether W54011 inhibits periodontitis has not yet been fully elucidated. To address this, we have investigated the probable anti-inflammatory mechanism of W54011 in LPS-treated inflammation in human gingival fibroblasts (HGFs). Methodology HGFs were isolated from healthy gingival tissue samples using the tissue block method and were identified with immunofluorescence staining. The CCK8 assay and reverse transcription-PCR (RT-PCR) were used to select the optimal induction conditions for Lipopolysaccharide (LPS) and C5aRA (according to supplementary data S1, S2 and S3). The levels of inflammatory cytokines, C5aR, and the activation of NF-κB/MAPK signaling pathways were determined by RT-quantitative PCR (RT-qPCR) and Western blotting. Results Immunofluorescence results showed that vimentin and FSP-1 were positive in HGFs and Keratin was negative in HGFs. Immunofluorescence staining demonstrated that C5aRA inhibited LPS-stimulated nuclear translocation of p-p65. RT-qPCR and Western blotting showed that C5aRA reduced the expression of IL-1β, IL-6, TNF-α, C5aR, p-p65, p-IκBα, p-JNK, p-c-JUN, and TLR4 in LPS-induced HGFs. Conclusion These findings suggested that C5aRA attenuated the release of inflammatory cytokines in LPS-induced HGFs by blocking the activation of the NF-κB and MAPK signaling pathways.

Abstract This study aimed to investigate the effects of eugenol (Eu) at different concentrations (0, 10, 50, 100, 200, and 300 μM/g protein) on the gel properties and chemical structure changes of porcine myofibrillar protein (MP). The results showed that Eu inhibited the increase of surface hydrophobicity and carbonyl content but did not prevent the oxidation-induced loss of thiol groups. Moreover, Eu intensified the loss of the α-helical conformation as well as the tertiary structure of MP under oxidative stress. The physicochemical changes at 10 and 50 μM/g Eu resulted in a significant enhancement of the gelling ability of MP and enhanced the positive role of oxidation in building elastic gel networks. Conversely, Eu at the concentrations of 100, 200, and 300 μM/g was not conducive to gelling ability, especially at 300 μM/g, and these concentrations were associated with Eu-induced protein conformational changes.

Abstract Objective To compare the clinical features, risk factors, distribution of Benign Paroxysmal Positional Vertigo (BPPV) subtypes, and effectiveness of canalith repositioning between geriatric and non-geriatric patients with BPPV. Methods A total of 400 patients with BPPV were enrolled. Canalith repositioning was performed according to the semicircular canals involved. Patients were divided by age into a geriatric group (≥60 years) and a non-geriatric group (20-59 years). Clinical characteristics, potential age-related risk factors, distribution of subtypes, and effectiveness of canalith repositioning were compared between the groups. Results Female sex was significantly more common in all age groups, with a peak female-to-male ratio of 5.1:1 in the group aged 50-59 years. There was a higher proportion of men in the geriatric group. A history of disease associated with atherosclerosis was significantly more common in the geriatric group (p< 0.05). Migraine was significantly more common in the non-geriatric group (p= 0.018), as was posterior canal BPPV. The horizontal canal BPPV (especially horizontal canal BPPV-cupulolithiasis), and multicanal BPPV subtypes were more common in the geriatric group, whereas anterior canal BPPV was more common in the non-geriatric group. Two canalith repositioning sessions were effective in 58.0% of the geriatric cases and in 72.6% of the non-geriatric cases (p= 0.002). There was a tendency for the effectiveness of canalith repositioning to decrease with increasing age. Conclusion BPPV was more common in women. However, the proportion of men with BPPV increased with age. Elderly patients often had a history of diseases associated with atherosclerosis (i.e., hypertension, diabetes, and hyperlipidemia). The horizontal canal BPPV (particularly horizontal canal BPPV-cupulolithiasis) and multicanal BPPV subtypes were more common and the anterior canal BPPV subtype was less common in elderly patients. The effectiveness of canalith repositioning may decrease with age. Therefore, older patients should receive more comprehensive medical treatment. Level of evidence: 4.

Regular exercise reduces the risk of malignancy and decreases the recurrence of cancer. However, the mechanisms behind this protection remain to be elucidated. Natural killer (NK) cells are lymphocytes of the innate immune system, which play essential roles in immune defense and effectively prevent cancer metastasis. Physical exercise can increase the activity of NK cells. Interleukin-15 (IL-15) is the best-studied cytokine activator of NK cells, and it was shown to have many positive functional effects on NK cells to improve antitumor responses. The aim of this study was to clarify the possible important mechanisms behind endurance exercise-induced changes in NK cell function, which may be highly correlated with IL-15. An animal model was used to study IL-15 expression level, tumor volume, cancer cell apoptosis, and NK cell infiltration after treadmill exercise. Although IL-15 was highly expressed in skeletal muscle, treadmill exercise further elevated IL-15 levels in plasma and muscle (P<0.05). In addition, tumor weight and volume of tumor-bearing mice were decreased (P<0.05), and liver tumor cell apoptosis was increased after 12 weeks of treadmill exercise (P<0.05). NK cell infiltration was upregulated in tumors from treadmill exercise mice, and the level of interferon-gamma (IFN-γ) and IL-15 were higher than in sedentary mice (P<0.05). The study indicated that regular endurance training can reduce cancer risk, which was related to increased IL-15 expression, activation of the immune killing effect of NK cells, and promotion of tumor cell apoptosis, which can ultimately control tumor growth.

ABSTRACT The objective of this study was to investigate the effect of cottonseed peptides from in vitro degradation with protease, Enterococcus faecalis ( E. faecalis ), or both on the growth performance, nutrient digestibility, and health status of pigs. We used four diets containing non-degraded, protease-degraded, E. faecalis -degraded, or both-degraded cottonseed meal at 20% for growing phase (6 wk) and 16% for finishing phase (7 wk), corresponding to barrows, with six replicates of six pigs each. Results showed that the in vitro degradation of cottonseed meal increased proportions of peptides and organic acids and decreased nitrogen-free extract and phytate phosphorus. The degraded cottonseed meal increased feed intake, body weight gain, feed efficiency, apparent ileal digestibility of dry matter, energy, crude protein, Ile, Leu, Lys, Met, Cys, Ser, and Gly, and blood profiles of superoxide dismutase, glutathione peroxidase, catalase, IgA, and IgG. The diet containing cottonseed meal by dual degradation showed more pronounced effects on nutritional compositions, growth performance, nutrient digestibility, and serum parameters than the other diets. The data suggest that cottonseed meal dually degraded in vitro with enzymes and probiotics can improve its nutritional properties and application in animal feed.

Abstract: Background: It is challenging to make informed decision in genetic counseling. Shared decision-making provides a chance in balancing the information and preferences between counselors and counselees. However, the status and prescriptions of shared decision-making have not been extensively studied in genetic counseling. Aim: To develop an up-to-date literature review of the shared decision-making in genetic counseling, identify knowledge gaps, and provide inspiration and suggestions for the development and practice of genetic counseling. Methods: “Genetic Counseling” and “Shared decision-making” were used to search in PubMed, Web of Science, Embase, Wanfang, CNKI and CBM databases. The search deadline was March 26, 2021. Results: A total of 22 articles were included, and four themes were identified: how people involved in genetic counseling, different stakeholders involved in SDM, multiple facilitators and barriers to SDM and the effectiveness of SDM in genetic counseling. Conclusion: Each participant needs to recognize their boundaries and make the best effort to involve in the shared decision-making. In the future, multinational studies should be considered for bringing shared decision-making to the global scale and well-designed studies are required to explore the long-term impact of shared decision-making in genetic counseling.

Resumen: Antecedentes: Es un reto tomar una decisión informada en el asesoramiento genético. La toma de decisiones compartida ofrece una oportunidad para equilibrar la información y las preferencias entre los asesores y los pacientes. Sin embargo, el estado y las prescripciones de la toma de decisiones compartida no se han estudiado ampliamente en el asesoramiento genético. Objetivo: Desarrollar una revisión bibliográfica actualizada de la toma de decisiones compartida en el asesoramiento genético, identificar las lagunas de conocimiento y proporcionar inspiración y sugerencias para el desarrollo y la práctica del asesoramiento genético. Métodos: “Genetic Counseling” y “Shared decision-making” se utilizaron para buscar en las bases de datos PubMed, Web of Science, Embase, Wanfang, CNKI y CBM. La fecha límite de búsqueda fue el 26 de marzo de 2021. Resultados: Se incluyó un total de 22 artículos y se identificó cuatro temas: cómo se involucran las personas en el asesoramiento genético, las diferentes partes interesadas involucradas en la GDS, los múltiples facilitadores y barreras para la GDS y la efectividad de la GDS en el asesoramiento genético. Conclusión: Cada participante necesita reconocer sus límites y hacer el mejor esfuerzo para involucrarse en la toma de decisiones compartida. En el futuro, se debe considerar la realización de estudios multinacionales para llevar la toma de decisiones compartida a la escala global y se requieren estudios bien diseñados para explorar el impacto a largo plazo de la toma de decisiones compartida en el asesoramiento genético.

Resumo Antecedentes: Tomar uma decisão informada em aconselhamento genético é um desafio. A tomada de decisão compartilhada (SDM) fornece uma oportunidade para contrabalançar a informação e preferências entre conselheiros e aconselhados. Entretanto, o status e prescrições da tomada de decisão compartilhada não têm sido extensivamente estudados em aconselhamento genético. Objetivo: Desenvolver uma revisão de literatura atualizada sobre a tomada de decisão compartilhada em aconselhamento genético, identificar lacunas no conhecimento e fornecer inspiração e sugestões para o desenvolvimento e prática do aconselhamento genético. Métodos: “Genetic Counseling” e “Shared decision-making” foram usados para pesquisar nos bancos de dados PubMed, Web of Science, Embase, Wanfang, CNKI e CBM. A data limite para a pesquisa foi 26 de março de 2021. Resultados: Um total de 22 artigos foram incluídos e quatro temas foram identificados: como pessoas se envolveram em aconselhamento genético, diferentes interessados envolvidos em SDM, facilitadores e barreiras múltiplas à SDM e efetividade da SDM em aconselhamento genético. Conclusão: Cada participante necessita reconhecer seus limites e fazer os melhores esforços para se envolver em tomada de decisão compartilhada. No futuro, estudos multinacionais devem ser considerados para trazer a tomada de decisão compartilhada a uma escala global e estudos bem desenhados são requeridos para explorar o impacto a longo prazo da tomada de decisão compartilhada em aconselhamento genético.

Abstract Allium mongolicum Regel is a desert plant that shows great resistance to wind erosion, drought, and low temperatures and is widely distributed in the desert lands of China. Understanding the molecular mechanisms underlying its drought resistance is essential for uncovering drought resistance genes and improving its beneficial traits. Here, a de novo RNA-Seq assembly analysis was conducted using the roots of 1-month-old seedlings under drought stress. Using pairwise comparisons of untreated plants (CK), 48-hour drought plants (G48h), and 96-hour drought plants (G96h), in total, 2,211 differentially expressed unigenes (DEGs) were obtained. Furthermore, using functional annotation, these DEGs were mainly involved in the plasma membrane, photosynthesis, and secondary carbohydrate metabolism. Moreover, genes involved in the ABA-mediated signaling pathway and secondary metabolism were upregulated in the roots. These results suggest that desert plants may use signaling and secondary metabolic pathways as adaptive responses to drought stress. Collectively, this work could help elucidate the molecular mechanisms underlying the ability of A. mongolicum Regel to respond to drought stress and aid in the selection of novel drought tolerance genes.

Resumen Allium mongolicum Regel es un tipo de planta desértica que muestra una gran resistencia a la erosión eólica, la sequía y las bajas temperaturas; se distribuye ampliamente en las tierras desérticas de China. Comprender los mecanismos moleculares subyacentes a la resistencia a la sequía de A. mongolicum Regel es esencial para descubrir genes resistentes a la sequía y mejorar este rasgo beneficioso en otras plantas. Aquí, el análisis de ensamblaje de ARN-Seq de novo se llevó a cabo utilizando las raíces de plántulas de 1 mes de edad en respuesta al estrés por sequía. Utilizando comparaciones por pares de plantas no tratadas (CK), plantas de 48 horas (G48h) y plantas de 96 horas (G96h), se obtuvieron un total de 2.211 unigenes (DEGs) expresados diferencialmente. Mientras tanto, usando la anotación funcional, estos DEGs fueron implicados principalmente en la membrana de plasma, la fotosíntesis, y el metabolismo de carbohidrato secundario. Por otra parte, los genes implicados en el camino ABA-mediado de la señalización y en metabolismo secundario para arriba fueron regulados para arriba en raíces. Estos resultados sugieren que las plantas del desierto pueden utilizar la señalización y las vías metabólicas secundarias como respuestas adaptativas al estrés por sequía. Colectivamente, este trabajo ayudará a comprender los mecanismos moleculares subyacentes a la capacidad de A. mongolicum Regel para responder al estrés por sequía y ayudar en la selección de nuevos genes de tolerancia a la sequía.

Abstract Atrial fibrillation (AF) represents the most common type of sustained cardiac arrhythmia in humans and confers a significantly increased risk for thromboembolic stroke, congestive heart failure and premature death. Aggregating evidence emphasizes the predominant genetic defects underpinning AF and an increasing number of deleterious variations in more than 50 genes have been involved in the pathogenesis of AF. Nevertheless, the genetic basis underlying AF remains incompletely understood. In the current research, by whole-exome sequencing and Sanger sequencing analysis in a family with autosomal-dominant AF and congenital patent ductus arteriosus (PDA), a novel heterozygous variation in the PRRX1 gene encoding a homeobox transcription factor critical for cardiovascular development, NM_022716.4:c.373G>T;p.(Glu125*), was identified to be in co-segregation with AF and PDA in the whole family. The truncating variation was not detected in 306 unrelated healthy individuals employed as controls. Quantitative biological measurements with a reporter gene analysis system revealed that the Glu125*-mutant PRRX1 protein failed to transactivate its downstream target genes SHOX2 and ISL1, two genes that have been causally linked to AF. Conclusively, the present study firstly links PRRX1 loss-of-function variation to AF and PDA, suggesting that AF and PDA share a common abnormal developmental basis in a proportion of cases.

Abstract The peeling process of fresh ginger during factory production causes not only waste but also pollution to the environment. Now, by fermenting whole pieces of fresh ginger into black ginger, people are able to further enhance the nutritional value of ginger while lowering the burden on the environment. In this experiment, fresh ginger was fermented into black ginger, and superoxide dismutase (SOD) crudely extracted from both fresh ginger and black ginger were gavaged separately in type 2 diabetic rats to compare the SOD activity and effect in rat liver during the treatment of type 2 diabetes. It was found that the SOD activity of extracts from fermented black ginger was much higher than that of extracts from fresh ginger. In addition, the activity of SOD in the liver of rats gavaged with black ginger extract is higher than that in the liver of rats gavaged with fresh ginger extract, which shows that black ginger extract has certain protective effects on the liver.

Abstract Melatonin is known to influence various physiological processes, including plant ripening; however, the effect of melatonin on postharvest blueberry fruit softening is unknown. Herein, the effects of melatonin at different concentrations (100, 300, and 500 μmol L-1) on the quality parameters, cell wall substances, cell-wall-degrading enzymes, and cell wall ultrastructure of blueberry fruits were investigated during low-temperature storage. Melatonin treatment was found to effectively delay the decline in fruit firmness, slow the loss of soluble solids and titratable acids, and decrease the respiration rate and decay rate of blueberry fruits. The blueberry fruits treated with 300 μmol L-1 melatonin exhibited the best quality. Melatonin treatment promoted anthocyanin accumulation, delayed ascorbic acid degradation, maintained a low relative membrane permeability in pulp, inhibited increases in lipoxygenase (LOX) activity, and reduced membrane lipid peroxidation. Moreover, melatonin controlled the formation of soluble pectin by inhibiting the activities of pectin methylesterase (PME), polygalacturonase (PG), cellulase (Cx), and β-glucosidase (β-Glu) while maintaining high contents of protopectin and cellulose. These results indicate that melatonin can effectively improve blueberry fruit quality and antioxidant capacity as well as and delay fruit softening by inhibiting the degradation of cell wall substances.

Abstract Introduction The standard approach to thyroidectomy is a collar incision via the anterior neck, and the neck scar has always been a source of worry for patients. Acceptable wound cosmetology has become a focus for thyroid surgeons. Objective To verify the effectiveness and cosmetic results of thyroidectomy through a lateral supraclavicular incision. Methods 180 patients were randomly divided into two groups: a lateral supraclavicular approach and a conventional transcervical approach. The main outcomes included incision length, intraoperative blood loss, operative time, total drainage volume, hospitalization expense, early postoperative pain measured by visual analog scale, infection, and perceived cosmetic outcome. Results There were no statistical significances between the two groups in terms of age, gender, nodule size, intraoperative blood loss, operative time, total drainage volume, hospital expense and postoperative complications, whereas there were significant differences in terms of incision length (5.2 ± 1.04 cm vs. 6.9 ± 1.14 cm, p < 0.05). Conclusions The lateral supraclavicular incision is a safe and feasible approach for thyroidectomy. Compared with conventional approach, it provides a better cosmetic result.

Resumo Introdução A abordagem padrão para tireoidectomia é uma incisão em colar na face anterior do pescoço; a cicatriz no pescoço sempre foi uma fonte de preocupação para os pacientes; consequentemente, a cosmetologia aceitável da ferida tornou‐se um foco de atenção para os cirurgiões de cabeça e pescoço. Objetivos Verificar a eficácia e os resultados cosméticos da tireoidectomia por meio de incisão supraclavicular lateral. Método Foram divididos aleatoriamente 180 pacientes em dois grupos: um grupo supraclavicular lateral (Grupo LS) e outro transcervical convencional (Grupo TC). Os desfechos principais incluíram comprimento da incisão, perda de sangue intraoperatória, tempo cirúrgico, volume total de drenagem, despesas hospitalares, dor no pós‐operatório imediato medida através de escala visual analógica, infecção e resultado cosmético percebido. Resultados Não houve significância estatística entre os dois grupos em termos de idade, sexo, tamanho do nódulo, perda sanguínea intraoperatória, tempo cirúrgico, volume total de drenagem, custo hospitalar e complicações pós‐operatórias, mas houve diferença significante em termos de comprimento da incisão (5,2 ± 1,04 cm vs. 6,9 ± 1,14 cm, p < 0,05). Conclusão A incisão supraclavicular lateral é uma abordagem segura e viável para tireoidectomia. Em comparação com a abordagem convencional, oferece um melhor resultado cosmético.

OBJECTIVES: To evaluate the efficacy of autologous platelet-rich plasma (PRP) gel in the treatment of refractory pressure injuries and its effect on wound healing time and quality of life of patients. METHODS: A random number table method was used to group 102 patients with refractory pressure injuries into either a control group (CG) (51 cases) receiving negative pressure wound therapy (NPWT) or a study group (SG) (51 cases) receiving NPWT+PRP gel. RESULTS: The total efficacy rate in the SG (92.16%) was higher than that in the CG (76.47%) (p<0.05). The SG exhibited lower visual analog scale (VAS) scores and pressure ulcer scale for healing (PUSH) scores, smaller wound sizes and depths, and shorter wound healing times than the CG after 21 days of treatment (p<0.05). After 6 months of treatment, the SG scored higher than the CG on the psychological, physiological, social functions, and daily activity domains on the World Health Organization Quality of Life (WHOQOL-BREF) scale (p<0.05). The incidence of postoperative complications in the SG (13.73%) was not significantly different from that of the CG (7.84%) (p>0.05). CONCLUSION: In the treatment of refractory pressure injuries, PRP gel can accelerate wound healing, reduce wound pain, shorten the treatment cycle, regulate tissue inhibitor matrix metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-9 (MMP-9) levels and the expression of specific proteins in granulation tissue, reduce the levels of the inflammatory factors interleukin-1β (IL-1β), IL-8, and tumor necrosis factor-α (TNF-α), and improve the quality of life of patients without increasing complications.

OBJECTIVES: Extracellular vesicle microRNAs (EV-miRNAs) have been demonstrated to be reliable candidate biomarkers for clinical applications. However, the clinical application potential of serum EV-miR-215-5p for gastric cancer (GC) remains poorly understood. The goal of our study was to determine the efficacy of serum EV-miR-215-5p in predicting the prognosis of GC. METHODS: Blood samples were collected from 118 patients with GC, 60 patients with benign gastric disease and BGD and 70 healthy controls. The relative levels of serum EV-miR-215-5p were measured using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Compared to patients with BGD and normal controls, GC patients exhibited remarkably higher serum EV-miR-215-5p level, especially those with early tumor recurrence (ETR). Receiver operating characteristic (ROC) curve analysis showed that serum EV-miR-215-5p was able to distinguish GC patients from BGD patients or healthy controls and GC patients with ETR from those without ETR. In addition, increased serum EV-miR-215-5p levels were notably correlated with invasive depth, TNM stage, and lymph node metastasis. Moreover, serum EV-miR-215-5p levels were greatly decreased after surgical treatment, but increased at the time of ETR. Survival analysis showed that patients with higher serum EV-miR-215-5p had shorter survival. Furthermore, serum EV-miR-215-5p was an independent risk factor for GC. CONCLUSIONS: Serum EV-miR-215-5p might be a novel biomarker for predicting ETR and prognosis of GC.