Halanaerobium, Psychrobacter, Photobacterium, and Tetragenococcus. Functional annotation revealed 15 pathways related to amino acid metabolism, including alanine, aspartate, glutamate, and histidine metabolism; lysine degradation; and arginine biosynthesis. This study demonstrated the approaches to identify microbiota functions and metabolic pathways, thereby providing a theoretical basis for taste formation mechanisms during traditional fish sauce fermentation.Complex microbial metabolism is key to taste formation in high-quality fish sauce during fermentation. To guide quality supervision and targeted regulation, we analyzed the function of microbial flora during fermentation based on a previously developed metagenomic database. The abundance of most identified genes involved in metabolic functions showed an upward trend in abundance during fermentation. In total, 571 proteins extracted from fish sauce at different fermentation stages were identified. These proteins were mainly derived from  Traditional fish sauce is a widely used liquid condiment prepared from low-value aquatic products through natural fermentation for 10–18 months , 3. The With the rapid development of systems biology techniques, it has become possible to study the microbial community structure, functional genes, and functional proteins in fish sauce fermentation systems, assisted by high-throughput multi-omics technology. Metaproteomics is less restrictive for studying microbial populations, as labeling of identified proteins is not required. Moreover, the microbial functional gene dataset constructed via metagenomics can help identify protein types, obtain microbial sources, infer amino acid sequences, and track complex functions of the microbial population in the environment. Metaproteomics has been applied to study various fermented foods, such as soybean paste , Pu-erh In this study, we analyzed the function of microbial flora involved in traditional fish sauce fermentation based on the metagenomic database established in our previous study . FurtherThe cap of the sampling bottle was opened atop an aseptic ultra-clean workbench, and tweezers were used to pick an alcohol-soaked degreased cotton wad to wipe the inside as well as the surface of the bottle and the cap repeatedly, ensuring that the entire surface of the bottle was wiped with alcohol. The bottle and the bottle cap were placed on the ultra-clean workbench, and a UV lamp was turned on for sterilization and irradiation for 20 min. The bottle was closed tightly with the cap after the alcohol had evaporated; the bottle was then labeled with the location, date, and time of sample collection and other information.Fish sauce samples were collected from a processing factory located in Guangdong, China, as per a previously reported method . AnchovyTraditional fermented fish sauce samples were collected from the top 10–12 cm of the upper surface of the fermentation broth at different fermentation stages. A total of five fermentation tanks were used for sampling in order to reduce sample error and ensure the stability of the experiment. The samples were collected after stirring. We collected 10.0 ± 0.2 L fermentation broth after 1, 3, 6, 9, and 12 months fermentation, respectively; hereafter, termed as 1M, 3M, 6M, 9M, and 12M, respectively. The sample replicates at each time point were well mixed on an ultraclean bench. The mixed samples collected at the same time point were defined as one sample. After collection, fermentation broth samples were filtered using qualitative filter paper  and collected in aseptic sampling bottle. The filtrate was divided into three biological replicates and stored in a refrigerator at −80°C until further analysis.Metagenomic DNA was extracted using the OMEGA DNA kit  according to the manufacturer’s instructions. Then, the DNA quality then was assessed on 1% agarose gels. The DNA sample was fragmented to a size of 300 bp using the M220 nucleic acid shea . A paired-end library was constructed using the TruSeq™ DNA sample prep kits . The libraries were sequenced on the Illumina Hiseq 2500 platform , 14.1, and nucleic acid sequences of genes longer than 60 bp were selected  within contigs were predicted using MetaProdigalselected . The Leaselected .The filtered sample (8 mL) was centrifuged at 15,000 × g for 5 min. After centrifugation, the supernatant was removed and purified by desalting on a PD-10 purified desalting column, and the effluent with protein components was collected after purification and desalting. The protein was precipitated by adding TAC/acetone solution (30 mL) for 12 h; the pellet obtained during the overnight precipitation was centrifuged at 10,000 rpm for 5 min to collect the protein precipitate, which was washed with acetone, resuspended in 8 M urea lysate, and sonicated . After ultrasonication, the solution was centrifuged at 12,000 rpm for 10 min, and the supernatant was collected as a protein solution, 60 μL of which was collected in a centrifuge tube. Dithiothreitol solution  was added to this tube, and the mixture was shaken on a shaker and incubated at 37°C for 1 h in a water bath. Subsequently, iodoacetamide solution  was added to the mixture, which was then vortexed and allowed to stand at 25°C for 1 h under dark conditions. All processed protein samples were transferred to an ultrafiltration tube, and the supernatants were discarded after centrifugation. Thereafter, UA solution  was added to the ultrafiltration tube, and the mixture was centrifuged; this step was repeated twice. Ammonium bicarbonate solution  was then added to the mixture, which was then centrifuged, and the supernatant was collected. This procedure was repeated three times. After replacing the collection tube, trypsin was added to the ultrafiltration tube at a protein:enzyme ratio of 50:1, followed by enzymatic hydrolysis at 37°C for 12–16 h  and Proteome Discoverer 1.4  programs were used for library identification and quantitative analysis. A protein dataset of macrogen unigene translation of traditional fish sauce samples was used as the database. The RAW files were submitted to the Mascot server through Proteome Discoverer; the established metagenomic database of fish sauce was selected, and a database search was performed. The data are available from ProteomeXchange under identifier PXD031089.2 and mapped to pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database.3 Enrichment analyses were performed based on Fisher’s exact test. Only functional categories and pathways with a p < 0.05 were considered significant. All analyses were performed with three biological replicates. Heat maps of hierarchical clustering were constructed using TBtools.4The FASTA protein sequences of differentially expressed proteins were analyzed using BLAST against the online eggNOG databaseThe statistics of the spliced and assembled sequences during fish sauce fermentation are shown in Traditional fish sauce fermentation is conducted in an open fermentation environment containing many microorganisms that cannot be cultivated and have not been studied. Based on the eggNOG database, a functional annotation statistical analysis of fish sauce during fermentation was performed, and the results are shown in To further determine the signal transduction and biochemical metabolic pathways involved in protein fermentation in traditional fish sauce, the difference in the proteome of the fish sauce was investigated using label-free MS/MS to explore the protein profiles related to taste compound formation. In total, 571 proteins were identified at different fermentation stages after the data were filtered at a false positive rate ≤ 0.01 . During By analyzing the homologous classification of proteins, the 571 proteins identified were classified using eggNOG functional annotation . Among tAspergillus oryzae participate in the decomposition of these substances. Among them, dipeptidase, dipeptidyl aminopeptidase, and leucine aminopeptidase play positive roles in promoting the formation of flavor during soy sauce fermentation  and 5′-inosine monophosphate (5′-IMP) , 30. Purpathways , and thepathways . Purine pathways . During o stages . The firo stages , 34.Amino acids are not only contributors of the nutrients and taste substances of traditional fermented foods but are also important precursors for flavor formation . TraditiHalanaerobium, Halococcus, and Halobacterium.The umami amino acid, glutamic acid, mainly converts carbohydrates to pyruvate through the EMP pathway; pyruvate is then converted to α-ketoglutarate through the TCA cycle . The bioHalanaerobium, Psychrobacter, Vibrio, and Halococcus. Another sweet amino acid, threonine, also uses aspartic acid as a precursor. Aspartic acid is catalyzed by aspartate kinase (EC: 2.7.2.4) to generate phosphoryl aspartic acid. Aspartate-semialdehyde dehydrogenase (EC: 1.2.1.11) reduces phosphoryl aspartic acid to aspartic semialdehyde and then generates threonine through the catalytic action of homoserine dehydrogenase (EC: 1.1.1.3), homoserine kinase (EC: 2.7.1.39), and threonine synthase (EC: 4.2.3.1). In addition, there are many other metabolic pathways involved in the production of threonine during the traditional fish sauce fermentation, and most enzymes in the metabolic pathway are covered in the annotation results.The sweet amino acid lysine uses the umami amino acid aspartic acid as a precursor, and the biosynthetic pathway is shown in Halanaerobium, Psychrobacter, Photobacterium, and Tetragenococcus based on the constructed microbial functional gene dataset. Functional annotation showed 15 pathways related to amino acid metabolism, including alanine, aspartate, and glutamate metabolism, histidine metabolism, lysine degradation, and arginine biosynthesis. The composition of taste compounds largely depended on the metabolism of the microbial flora. The biosynthetic pathways of various amino acids were different, but the formation of their carbon skeletons, mainly via the citric acid cycle and the pentose phosphate pathway, was common. This study provided a detailed evaluation of fish sauce fermentation, enabling the development of better strategies for targeting regulation and quality monitoring of fish sauce.The current study sheds light on the understanding of microbiota and metabolic pathways involved in Chinese fish sauce fermentation. The results showed an upward trend in the abundance of most identified genes predicted to be involved in metabolic functions during fermentation. In total, 571 proteins extracted from fish sauce at different fermentation stages were identified after screening and filtering. The protein composition of the traditional fish sauce during fermentation changed significantly. The results demonstrated that the proteins in traditional fish sauce were derived from http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD031089.The original contributions presented in the study are publicly available. This data can be found here: YWa: writing—original draft, conceptualization. YWu and LL: conceptualization. CL: writing—review and editing. YZ, XY, and SC: visualization. HX, LS, and BQ: software and formal analysis. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Mild cognitive impairment (MCI), a reversible intermediate state, plays an important role in the development and prevention of dementia. The relationship between pulmonary function and MCI risk has not yet been well-elucidated.We included 2,947 rural Chinese residents aged ≥35 years who were free from a history of stroke, dementia, or other brain diseases and measured pulmonary ventilatory function using calibrated spirometry according to the recommended method. MCI was assessed with the Montreal Cognitive Assessment-Basic for Chinese scale. Logistic regression models and restricted cubic splines with covariate adjustment were performed to explore the association between pulmonary function and MCI risk.Pnon−linear = 0.032).The prevalence of MCI increased with decreasing pulmonary function, from the lowest quartile to the highest quartile of pulmonary function: 63.9, 50.5, 43.8, and 43.6%, respectively. After adjustment for confounding factors, participants in the first quartile had a significantly increased risk of MCI , with the highest quartile as the reference. In the subgroup analysis, a significant association of pulmonary function and MCI was found in females and those with low physical activity. Meanwhile, we observed an L-shaped relationship between pulmonary function and MCI (Poor pulmonary function was associated with an increased risk of MCI among rural Chinese adults, and presented a non-linear relationship. These findings remind us of the need for early cognitive assessment in local populations with lower pulmonary function. Dementia is one of the major public health problems and the leading cause of disability in elderly individuals worldwide . People In addition to well-recognized risk factors, impaired pulmonary function is becoming increasingly recognized as a predictor of cognitive performance . A recenAll data were collected from a baseline survey in rural areas of Fuxin County, North China, from June to August 2019. Based on the demographic characteristics, participants were recruited from the eastern, southern, and northern townships to take part in our survey. The inclusion criteria were residents aged 35 and above who resided locally for at least 5 years and could sign a consent form during the investigation. Subjects were excluded if one of the following criteria were met: (1) were pregnant; (2) developed severe liver and renal failure; or (3) were unwilling to participate in this study. Finally, 4,689 participants were recruited. All participants gave their informed consent at the time of the examination and the Committee of China Medical University authorized the study protocol. The procedures followed were performed under the ethical standards of the responsible committee on human experimentation of China Medical University ([2018]083).n = 1,289), MoCA scores (n = 227) and confounding factors (n = 84). In addition, 142 participants with a self-reported history of stroke, depression, brain disease or dementia were excluded. In the end, Ultimately, 2,947 participants were enrolled in the analysis. A flowchart of the specific inclusion and exclusion criteria for participants in the current analysis is shown in Of all 4,689 eligible participants, 1,600 dropped out due to missing information about spirometric indicators  according to the recommended method. Participants were asked to take a sitting position and put the disposable mouthpiece into the mouth, with lips sealed tightly around the mouthpiece, while pinching the nose, breathe calmly, then took the deepest breath possible to fill the lungs with air and blow as hard and as fast as possible . For eac studies , 16, we The Montreal Cognitive Assessment (MoCA), published in 2005, has been proven more sensitive and reliable in MCI screening than other scales . For theRelevant data on demographic variables , lifestyle factors , and history of disease were collected by face-to-face interview with a standardized questionnaire. Histories of disease were defined as self-reported and confirmed by medical records. Current smokers were defined as people who smoked at least one cigarette per day and continued for half a year. Alcohol ingestion was defined as at least three drinks every week for 6 months. Physical activity was rated three levels, low, moderate and high, based on the occupation engaged in.2. Hypertension was taken as an antihypertensive medication in the last 2 weeks, DBP ≥90 mmHg or SBP ≥140 mmHg /height (m)140 mmHg . Diabete140 mmHg .t-test for continuous variables or the chi-square test for categorical variables.Continuous variables are represented as the mean ± standard deviation (SD) or medians (interquartile ranges), and categorical variables are reported as frequencies (percentages). Analysis of variance or the Kruskal–Wallis rank-sum test for continuous variables, respectively, while the chi-square test for categorical variables was used to compare differences in basic characteristics of different pulmonary function quartile groups. In addition, baseline characteristics comparisons between MCI and non-MCI were also made using Student's We examined associations of each pulmonary function parameter  and MCI. Binary logistic regression models were used to estimate the odds ratios (ORs) and 95% confidence intervals (CIs) for composite pulmonary function and MCI. The composite pulmonary function score was assessed as quartiles 1–4 and quartile 1 vs quartiles 2–4. Three models were assessed, each building upon the prior model. Model 1 was unadjusted, model 2 was further adjusted for age and sex, and model 3 controlled for the variables in Model 2 plus body mass index (BMI), marital status, ethnicity, education level, smoke exposure, alcohol consumption, physical activity, hypertension, diabetes, histories of coronary heart disease (CHD), and chronic obstructive pulmonary disease (COPD). Tests for trends across quartiles were examined using ordinal values in separate models.z-score of composite pulmonary function distribution. The threshold level for each section was determined using likelihood-ratio tests and bootstrap resampling methods. We further applied a piecewise regression model to examine the specific relationship between composite pulmonary function and MCI.Moreover, we used restricted cubic splines (RCS) based on logistic regression models to visualize the relationship between pulmonary function and MCI on a continuous scale by adjusting the same confounders, with knots placed at the 10th, 50th, and 90th percentiles of the n = 176), histories of respiratory diseases, including COPD (n = 61), asthma (n = 4), pneumonia (n = 5), and other chronic lung diseases (n = 2). Subjects with occupational exposure (n = 176) or current smokers (n = 980) were also excluded.To assess relevant factors that had a modifying effect on the relationship between pulmonary function and MCI, we further conducted subgroup analyzes with study subjects stratified by sex, age (<65 or ≥65 years), smoking status (yes or no) and low physical activity (yes or no), respectively. We performed sensitivity analyzes to test the robustness of the results by repeating the analysis after excluding participants with respiratory symptoms (http://www.R-project.org/) and other statistical analyzes were performed using IBM SPSS statistical software version 25.0 . A 2-sided P value of <0.05 was considered statistically significant.The RCS analysis and threshold effect analyzes were conducted in R software, version 4.2.0 (P = 0.359).Out of 2,947 subjects with available data, 1,487 (50.5%) were observed to have MCI. The average age of the participants was 57.5 ± 9.8 years, and 1,939 (65.8%) were females. The characteristics of the study participants are described by different quartiles of pulmonary function in P < 0.001) in the fully adjusted model. When quartiles 2–4 were further combined, the participants in the lowest quartile had a 42.4% higher risk of MCI than those in the upper quartiles . Additionally, we observed a non-linear relationship between pulmonary function and the risk of MCI .There were inverse associations between FVC, FEV1 and PEF and the risk of MCI . Consistz-score for composite pulmonary function <0.214. As shown in P ≥ 0.05). Notably, we only found a relationship between lower pulmonary function and a higher risk of MCI in the female group and those with low physical activity, and the multivariate-adjusted ORs for MCI were 1.465  and 2.000 , respectively. The risk of MCI for individuals with lower pulmonary function in non-smoker was higher , and even higher in smokers .We further performed subgroup analyzes to explore potential modifier and interaction effects on the PF-MCI association in participants with After exclusion of participants with respiratory symptoms, histories of respiratory diseases, occupational exposure and those who were current smokers, sensitivity analyzes yielded no any substantial differences from the main results .In this population-based study of rural adults in Northeast China, we highlighted that poor pulmonary function was associated with a significantly increased risk of MCI and presented a non-linear relationship. Moreover, poor pulmonary function was associated with an elevated risk of MCI in females and those with low physical activity.In normal human lungs, airflow depends on the expiratory driving pressure including the magnitude of expiratory muscle and elastic recoil as well as the size and viscoelastic properties of the lung and airway . FVC and2 exhibited that lower pulmonary function during midlife was associated with an increased likelihood of MCI in later life (2)  . Indeed,life (2) . Residenlife (2) , 34. In via an intricate web of signaling involving hypoxia, oxidative stress, systemic inflammation, neurotransmitter function, or a combination of these processes . The patients/participants provided their written informed consent to participate in this study.YZ and LZ foresaw this work and designed the overall research. JX, WY, RL, HuG, HG, and WF collected primary data and screened the data. CG performed the statistical analysis and drafted the manuscript. MM participated in data interpretation and critically revised the manuscript. HaG supervised the analysis. All authors contributed substantially to discussions of the content of this paper and agreed to the submitted version of the manuscript.This research was supported by funds from the National Nature Science Foundation of China [No. 82073645].The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The outbreak of COVID-19 has become a global public health emergency, causing great psychological distress to nurses. It is unknown whether the pandemic will affect the work engagement of nurses, the relationship between psychological capital, job satisfaction, and work engagement among nurses, and whether there are differences between nurse specialists and general nurses during the pandemic.The purpose of this study was to compare psychological capital, job satisfaction, and work engagement among nurse specialists and general nurses during the pandemic, as well as to test the role of job satisfaction as a mediator in the association between psychological capital and work engagement among nurses, and to examine whether the underlying mechanism of the relationship between psychological capital and job satisfaction differs between nurse specialists and general nurses.A convenience sampling was used to assess a sample of 372 nurse specialists and 318 general nurses from nine provincial general hospitals in China to participate in the online survey. Data were collected using self-report questionnaires, including the following tools: self-designed socio-demographic questionnaire, psychological capital scale, job satisfaction scale, and work engagement scale.Compared with general nurses, the nurse specialists had higher psychological capital, job satisfaction, and work engagement. Job satisfaction partially mediated the positive association between psychological capital and work engagement and the indirect effect was stronger in nurse specialists in comparison to general nurses during the COVID-19 pandemic.The findings provide important practical implications for future intervention programs aimed at enhancing nurses’ work engagement, which may be realized through strengthening psychological capital and job satisfaction during the pandemic. Moreover, considering the cost-effectiveness of limited health care spending, nursing managers should pay more attention to the continuing professional development of young general nurses. With the rapid development of health care and the aging of the population, the demand for quality care continues to increase, which brings great challenges to nursing work. However, the World Health Organization estimates that a shortfall of 5.7 million nurses has been predicted by 2030 across the world . In ChinWork engagement is a positive state of mind associated with work that is characterized by vigor, dedication, and absorption . Vigor rAside from psychological capital, job satisfaction has become a supportive factor in work engagement . Job satIn light of a potential nursing shortage, the growing burden of diseases in the aging population and scarcity of health resources sharpen the need for a sustainable nursing health human resources strategy to satisfy the rising demand for care and maximize nursing efficiency , 31. TheTherefore, it is necessary to focus on the impact of the pandemic on nurses’ work engagement. And because specialist nurses and general nurses have different scopes of practice, it is important to explore whether there are differences in psychological capital, job satisfaction and work engagement between them, which may benefit in promoting the continuing professional development of nursing staff. In addition, little is known about the relationship between psychological capital, job satisfaction and work engagement among nurses during the pandemic. As a result, the purpose of our study is to compare psychological capital, job satisfaction, and work engagement among nurse specialists and general nurses during the pandemic, as well as propose the hypotheses that the relationship of psychological capital on work engagement is mediated by job satisfaction, and to test whether the underlying mechanism of the relationship between psychological capital and job satisfaction differs between nurse specialists and general nurses during the COVID-19 pandemic. A moderated mediation model  was consA multi-center cross-sectional survey was carried out from May to October of 2021 from 9 provinces in China. The researchers created an electronic questionnaire on the questionnaire star platform and sent it to nursing administrators in nine provincial general hospitals of China through WeChat, mainly including Liaoning, Jilin, Heilongjiang, Sichuan, Guangdong, Anhui, Henan, Anhui, Guangxi Province. They were asked to deliver the survey to the nurse through We Chat. Participants could scan the QR to read and submit the informed consent agreement and questionnaires. Each participant was only allowed to submit once in order to avoid double submission.The following were the eligibility criteria for nurse specialist enrollment: (a) hold a Chinese nursing specialist certificate after completing at least 6 months of training; (b) hold a Chinese registered nurse license; (c) work full-time; (d) have submitted a declaration of consent.The following eligibility criteria for general nurse were: (a) hold a Chinese registered nurse license; (b) work full-time; (c) have submitted a declaration of consent.The exclusion criteria for two group nurses were nursing staff who were not directly involved in patient care, such as absent due to sick leave, personal leave, study, or further training.The sample size calculator G*Power (3.0.10) was used . To idenAccording to the purpose of our study, we designed the general socio-demographic data, which mainly includes gender, age, health status, physical condition, marital status, family status, years of working, education, job title, department position, total night shifts per month and monthly income.The Chinese version of the Psychological Capital Scale was used to assess the psychological capital of nurses. Luthans et al.  developeThe job satisfaction scale of medical staff developed by Chinese scholar Wang et al.  was usedThe Chinese version of the Utrecht Work Engagement Scale was used to assess work engagement of nurses. It was developed by Schaufeli et al.  with ninTo ensure accuracy, two researchers who did not know the study design entered the data after all surveys were completed. They received training on how to check, input and code the data into IBM SPSS v23.0 before analyzing the data. Only if they pass the training exam can they participate in data entry.t-test was used to analyze and compare the psychological capital, job satisfaction, and work engagement of the nurse specialist group and the non-nurse specialist group. Third, Pearson’s or Spearman’s correlation analysis was used to analyze the correlation between social-demographic variables, psychological capital, job satisfaction and work engagement among nurses. To check for collinearity among the independent variables, both bivariate Pearson’s correlations and variance inflation factors were performed, with significant correlations less than 0.8 among these variables and variance inflation factors (VIFs) were < 10, indicating that multicollinearity was not a problem. Then, a simple mediation analysis of job satisfaction mediating the relationship between psychological capital and work engagement was tested using Hayes’s PROCESS macro for SPSS (Model 4)  . The indModel 4)  was utilModel 4) .P-value was two-tailed and we inferred statistical significance if α was < 0.05.According to the previous studies , age, se2 = 2686.43, degrees of freedom (df) = 170, χ2/DF = 15.803, comparative fit index (CFI) = 0.849, goodness of fit (GFI) = 0.676, normed fit index (NFI) = 0.841, root mean square error of approximation (RMSEA) = 0.147, which indicating that there is no significant common method variance problem in this study.The data for this study were collected through self-report, which may have been influenced by common method bias, reducing the validity of the results. In this study, we followed strict confidentiality and voluntarism principles and asked participants to be honest in their answers to each question. Data collectors are used to collect and enter data. These methods effectively control the bias of the common method. In addition, following the method suggested by Podsakoff et al. , common p < 0.001). In nurse specialists, 80.11% had children compared to 49.37% in general nurses, respectively (p < 0.001). 138 (37.1%) of nurse specialists had worked between 11 and 15 years, 104 (27.96%) had worked more than 21 years, and 86.48% of general nurses had worked fewer than 15 years. In terms of working years, there was a considerable difference between the two groups (p < 0.001). Among nurse specialists, 2.69% were junior nurses, 26.61% were senior nurses, 52.15% were nurses-in-charge and 18.55% were vice-director nurses and above, however, 18.90% were junior nurses, 51.90% were senior nurses, 23.90% were nurses-in-charge and 5.30% were Vice-director nurse and above in general nurses. There was a significant difference in the job title between the two groups (p < 0.001). Among the department position of nurse specialists and general nurses, there were 155 (41.67%) and 246 (77.36%) of general nurses, 55 (14.78%) and 27 (8.49%) of team teachers, 49 (13.17%) and 17 (5.35%) of nursing team leaders, 99 (26.61%) and 25 (7.86%) of head nurses, and 14 (3.76%) and 3 (0.94%) head nurses and above. There was a significant difference in department position, marital status and the number of night shifts per month between the two groups (p < 0.001), but no difference in educational level, health status and monthly income (p = 0.227).A total of 700 nurses took part in the study. After excluding 10 invalid questionnaires, a total of 690 valid questionnaires remained for an completion rate of 98.6%, which included 372 nurse specialists and 318 general nurses. Females predominated in both groups. In terms of age, 206 (55.4%) of nurse specialists were over 35 years old, while only 59 (18.56%) of general nurses were, indicating that most numbers of general nurses were younger than nurse specialists. The age difference between the two groups was significant (p < 0.05). The nurse specialists’ job satisfaction score (76.59 ± 12.20) was significantly higher than general nurses (74.44 ± 12.57). In terms of work itself, interpersonal relationship, and work return, the nurse specialists scored significantly higher than general nurses (p < 0.05). However, there was no significant difference in work pressure, working condition, and organizational management between the two groups (p > 0.05). The nurse specialists’ work engagement scores (39.22 ± 12.35) and vigor scores (13.20 ± 4.15) were significantly higher than general nurses (37.16 ± 12.65) and (12.21 ± 4.44). However, we found no statistically significant differences in dedication and absorption scores between the two groups (p > 0.05).As shown in n = 690). The score for work engagement was 38.27 (SD = 12.52). The scores for psychological capital and job satisfaction were 94.17 (SD = 16.82) and 75.60 (SD = 12.41), respectively. As shown in r = 0.776, p < 0.001; r = 0.629, p < 0.001). Moreover, psychological capital was found to be positively related to job satisfaction . The absolute values of the correlation coefficients between the three variables were less than 0.8, indicating a weak to moderate correlation between the variables and no multicollinearity problem ( problem .p < 0.05). Moreover, gender, years of working were all positively related to psychological capital, job satisfaction and work engagement, while total night shifts per month were negatively related to all three variables among general nurses (p < 0.05). In addition, the findings also showed positive correlations between psychological capital, job satisfaction, and work engagement in both groups (p < 0.001).p < 0.001). The mediating effect was then investigated using model 4 in the PROCESS macro. After accounting for nurses’ socio-demographic, psychological capital was found to be positively associated with job satisfaction , which in turn predicted work engagement . Job satisfaction also had a significant indirect effect , according to bootstrapping analyses. Furthermore, there was a significant direct relationship between psychological capital and work engagement . Therefore, job satisfaction partially mediated the relationship between psychological capital and work engagement. The mediating effect accounted for 16.34% of the total effect.According to the Hypothesis, we investigated if the relationship between psychological capital and work engagement would be mediated by job satisfaction . Before p < 0.05), indicating that the effect of psychological capital on job satisfaction was moderated by specialization. Thus, the moderated mediation model was established as the first stage of the mediation effect was moderated by specialization.The study hypothesized that specialization would moderate the effect of psychological capital on job satisfaction. As shown in simple = 0.866, p < 0.001), whereas for general nurses, the association between psychological capital and job satisfaction was still significant, but much smaller . The study plotted the association of psychological capital with job satisfaction separately for nurse specialists and general nurses to interpret interactions with moderating variables ] than for general nurses .This study compared the work engagement, psychological capital, and job satisfaction of nurse specialists and general nurses and to investigate the relationship between psychological capital and work engagement of nurses, as well as the role of job satisfaction in mediating the relationship between psychological capital and work engagement in China during the pandemic. We found that compared with general nurses, nurse specialists had higher work engagement, psychological capital and job satisfaction, psychological capital was positively correlated with work engagement. Job satisfaction partially mediated the positive association between psychological capital and work engagement. Specifically, the association between psychological capital and job satisfaction was stronger for nurse specialists compared to general nurses during the COVID-19 pandemic. Overall, the results explain the hypotheses proposed in this study.The study found that nurses showed a higher level of work engagement during the COVID-19 pandemic than Wan et al.  were befThe study found that nurse specialists had a higher level of work engagement than general nurses. This could be because the two groups of nurses differed in age, years of working, night shift, professional practice environment, and professional status (such as job title or department position), which was consistent with the previous study . Vigor rWe found that the psychological capital of nurse specialists was significantly higher than that of general nurses. On the four dimensions of self-efficacy, hope, resilience and optimism, the nurse specialists outperformed the general nurses. It could be due to the fact that self-efficacy is one’s belief in their ability to achieve their goals , which cThe study found that the job satisfaction of nurse specialists was higher than general nurses. This could be due to the fact that general nurses are mostly young nurses who had to shoulder the responsibility for coping with the pandemic in addition to their regular nursing duties. A previous study reported that of the 28,600 nurses recruited from across China to provide care to patients with COVID-19 in Hubei, 40% were younger than 30 years, which indicated young health-care providers were the backbone of the COVID-19 crisis in China . TherefoA previous study noted that experienced nurses were appreciated by students and young nurses for the knowledge and skills that helped ease their transition to the workplace, allowing them to better realize their roles within the healthcare team and their respective scopes of practice . HoweverOne of the key findings of the study was that psychological capital was positively associated with work engagement, which could be explained by the JD-R model theory  stated tAlthough the present study compared the work engagement between two groups of nurses and furthers our understanding of the mediating mechanism underlying the association between psychological capital and work engagement among Chinese nurses, several limitations need to be acknowledged. First, because this is a cross-sectional study, the causality of the associations between psychological capital, job satisfaction and work engagement cannot be inferred. However, we believe that our findings still provide useful and important information on the mediation model between psychological capital and work engagement. Future longitudinal study designs are needed to more robustly validate the causal relationship in this model. Second, the study data were derived from self-reports, which could lead to potential subjective and recall bias that could affect the accuracy of the assessment. Moreover, social expectations bias among participants should be considered. In this study conducted in the same hospitals setting, nurses may likely report similar work engagement results, as they felt it might make others feel that they had high levels of work engagement, particularly during the pandemic. To reduce this bias, researchers reassured all participants that there were no right or wrong answers and were informed to answer all questions based on their first instinct to minimize the impact of elucidating socially desirable responses. In addition, all participants were recruited from nine provinces in China. Therefore, the results may not apply to all nurses. Long-term studies that require random sampling in other organizational and cultural contexts should be considered to improve the generalizability of the results in the future.As evidence has shown that mental health support is urgently needed to help nurses be more productive all over the world in combating the COVID-19 pandemic to relieve their psychological distress . Our stuOur results suggest that compared with general nurses, nurse specialists had higher work engagement, psychological capital, and job satisfaction. More importantly, this study is primarily based on the JD-R model and considers the COR theory to explore that job satisfaction partially mediated the positive association between psychological capital and work engagement, and the indirect effect was stronger in nurse specialists in comparison to general nurses during the COVID-19 pandemic. Our study provides nursing managers with suggestions for team building and management that have theoretical and practical significance. Consider the cost-effectiveness of limited healthcare expenditures during the pandemic, nursing managers should pay more attention to the continuing professional development of young general nurses, ensuring sufficient training and mentorship for them in combating the pandemic by experienced nurse specialists, which helps to create a cohesive health care team during the pandemic. Second, our finding can help nursing managers correctly understand the mechanism of the relationship among psychological capital, job satisfaction and work engagement and adopt effective intervention strategies to promote nurses’ work engagement.The original contributions presented in this study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by the First Affiliated Hospital of China Medical University . The patients/participants provided their written informed consent to participate in this study.MZ and YLiu conceptualized and designed the project. MZ, HC, NW, and YLi acquired and managed the data and performed statistical and data analysis. MZ and HC drafted the manuscript. YLiu and XL revised the manuscript. All authors contributed to the article and approved the submitted version.


Sorting and assembly machinery component 50 homolog (SAMM50) gene single-nucleotide polymorphisms (SNPs) have been connected with the susceptibility of nonalcoholic fatty liver disease (NAFLD), but with inconsistent results across the current evidence. The present work was schemed to explore the association between SAMM50 gene SNPs and NAFLD vulnerability via meta-analysis.PubMed, Web of Science, Cochrane Library, China National Knowledge Infrastructure (CNKI), and Wanfang were retrieved for eligible literature previous to June 10, 2021. The odds ratios (ORs) of the dichotomic variables and the standardized mean difference of quantitative variables with corresponding 95% confidence intervals (95% CIs) were computed to evaluate the strength of the associations. The quality of included studies was assessed using Newcastle-Ottawa Scale (NOS).rs2143571, rs3761472, and rs738491 polymorphisms respectively. The pooled data revealed that the 3 polymorphisms had conspicuous associations with NAFLD susceptibility: rs2143571, A vs. G, OR=1.51, 95% CI, 1.37–1.66, P < .01; rs3761472, A vs. G, OR=1.50, 95% CI, 1.35–1.67, P < .01; rs738491, A vs. G, OR=1.51, 95% CI, 1.40–1.63, P < .01.In total, 8 case-control studies encompassing 6297 NAFLD patients and 7306 disease-free controls in this meta-analysis. Ultimately, this analysis included 8, 6, and 5 studies for rs2143571, rs3761472, and rs738491 polymorphisms of the SAMM50 gene are appreciably associated with augmented risk of NAFLD vulnerability. It will provide the latest evidence to support the susceptibility of SAMM50 gene polymorphisms and NAFLD, and provide strategies for the prevention and treatment of NAFLD.This meta-analysis suggests that  It is gradually considered the liver disease component of metabolic syndrome, which is a risk factor for further development of fatty liver, nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma.–4 The global burden of NAFLD is rapidly increasing and is expected to become the most common indication for liver transplantation with the alarming rise in the prevalence of obesity and metabolic syndrome. According to the World Gastroenterology Organization, NAFLD has been increasing during the past 20 years and is now one of the most common types of liver disease in Western countries. According to the present data, NAFLD affects 10%–30% of the general population in various countries, which has been viewed as a huge global health burden.,8Nonalcoholic fatty liver disease (NAFLD) is an assessment of fatty infiltration in the liver by histopathologic examination in the absence of excessive alcohol intake, autoimmune, viral infection, or drug-induced liver disease. At present, genome-wide association studies (GWAS) have demonstrated several conspicuous genetic susceptibility genes such as PNPLA3, SAMM50, PARVB, APOC3, PPAR-γ, NCANCILP2, FABP1, AGTR1, GSTs, which had been verified the crucial roles in the disease onset and progression of NAFLD.–15 Among them, rs738491, rs2143571, and rs3761472 in the sorting and assembly machinery component 50 homolog (SAMM50) gene were demonstrated to be notably associated with NAFLD susceptibility. Up to now, there have been several reports on SAMM50 gene SNPs and NAFLD susceptibility, but the results were inconsistent. Additionally, the real situation of the whole population cannot be reflected due to the limited sample size and regional and ethnic differences in the existing single studies. Therefore, a meta-analysis will be conducted on the existing research results to obtain more authentic and reliable results.Little is known about the latent mechanism involved in the development and pathogenesis of NAFLD, and yet it is a complicated metabolic process in which both environmental and genetic factors are etiology.This study depended on previously published literature and public databases. Thus, this work does not need ethical approval and patient consent. Two independent researchers searched the PubMed, Web of Science, Cochrane Library, CNKI, and Wanfang databases before June 10, 2021. The searching strategy of PubMed was exhibited as follows:  AND (“Sorting and assembly machinery component 50 homolog” [Mesh] OR SAMM50) AND (“Single Nucleotide Polymorphism” [Mesh] OR Variant OR SNP OR Polymorphism OR Mutant OR Mutation OR Variation). No language restriction was set. This study depended on previously published literature and public databases. Thus, this work does not need ethical approval and patient consent.Original studies were incorporated into this analysis in the light of the following inclusion criteria: (i) case-control studies; (ii) study investigated the associations of SAMM50 polymorphisms and NAFLD predisposition; (iii) NAFLD is diagnosed by pathology or ultrasound; (iv) control subjects were disease-free individuals; (v) detailed genotype data can be calculated for ORs and 95% CIs. Correspondingly, the exclusion criteria were as follows: reviews, conference abstracts, commentary articles, letters to the editor, animal studies, unpublished data, case reports, and secondary causes of steatosis, including alcoholism, total parenteral nutrition, hepatitis B/C virus infection, and drug usage.The Newcastle–Ottawa Scale (NOS) was performed to evaluate the methodological quality of included studies by 2 independent investigators. The NOS is composed of 3 aspects, namely selection, comparability, and exposure. Each study could receive 0 to 9 scores. Nevertheless, studies with ≥6 scores were regarded as high-quality studies. Disagreement was resolved by discussion.The extracted information contained the name of the first author, year of publication, country, ethnicity, numbers of NAFLD and control subjects, diagnostic methods of NAFLD, genotype frequencies, and Hardy–Weinberg equilibrium (HWE) results. Two investigators extracted the information independently. Summary ORs with 95% CIs were computed to specifically evaluate the relationship between SAMM50 gene polymorphisms and NAFLD susceptibility. Cochran’s Q-test and I2 test were applied to appraise the between-study heterogeneity. The random-effect model was employed to calculate merged ORs if P < .1, I2 > 50%. If not, the fixed-effect model was utilized for data synthesis. Sensitivity analysis was performed as well to assess the stability of all incorporated studies via the leave-one-out method. When the included studies were more than 10, a funnel plot was employed to evaluate the publication bias. On the contrary, less than 10 studies are not required. All analyses were done using Review Manager 5.3 software .In the case of polymorphism with 2 alleles (A1 and A2), both the NAFLD and disease-free participants could be classified into 4 exposure groups . Because the underlying genetic model for SAMM50 gene polymorphisms and NAFLD was unclear, the estimation of the association was based on 5 genetic models including allele contrast (A1 vs. A2), homozygous contrast (A1A1 vs. A2A2), heterozygous contrast (A1A2 vs. A2A2), dominant contrast (A1A1 + A1A2 vs. A2A2), and recessive contrast (A1A1 vs. A1A2 + A2A2). The control participants of incorporated studies were estimated via HWE.A primary search of electronic databases retrieved 26 potentially relevant publications: 13 from PubMed, 12 from Web of Science, 0 from Cochrane Library, 0 from CNKI, and 1 from Wanfang. No additional records were acquired from other sources. And then 20 studies remained after removing duplicated articles. Subsequently, a total of 4 irrelevant articles were excluded based on titles and abstracts screening. After applying inclusion and exclusion criteria, 3 unrelated articles (not related to NAFLD/SAMM50 gene polymorphisms), and 3 reviews were removed and 2 full-text article was not available. Ultimately, 8 studies  went into the process of meta-analysis. Overall, a flowchart summarizing the procedure of literature identification was illustrated in Figure –23 India, China, USA, Mexico, and Korea. They were all case-control designed and published between 2012 and 2018. It is worth noting that 2 studies  were genome-wide association studies (GWAS), including 2 stages of discovery I and validation II. In total, 8 studies encompassing 6297 cases and 7306 control participants were analyzed in the current analysis. For rs2143571, rs3761472, and rs738491 polymorphisms, there were 8, 6, and 5 studies ultimately incorporated, respectively. The main characteristics of the included literature were shown in Table  and Kanth et al did not provide the genotype frequency of the control group, so HWE could not be calculated. Studies with≥6 scores were regarded as high-quality studies according to the evaluation of methodological quality , homozygous model , heterozygous model , dominant model , and recessive model   including 6297 NAFLD patients and 7306 controls for rs3761472 to perform data analysis. The allelic model was analyzed by a random-effect model owing to significant heterogeneity observed. As the heterogeneity of genetic models was not significant, the fixed-effect model was employed for data analysis. The pooled data indicated there was a noticeable association between rs3761472 polymorphism and NAFLD susceptibility under allelic model , homozygous model , heterozygous model , dominant model , and recessive model  , homozygous model , heterozygous model , dominant model , and recessive model  (Table A total of 5 studies with 2150 cases and 4792 controls were used for data pooled. The fixed-effect model was employed for data analysis owing of the between-study heterogeneity was not significant. The results indicated an obvious association between P value, ORs, and 95% CIs did not change substantially. Therefore, the outcomes were considered to be statistically robust and reliable. The funnel plot for assessment of publication bias was not implemented on account of fewer than 10 research.After the omission of an individual study, the recalculated  In addition, SNPs in humans might be one of the critical steps to disclosing the genetic factor for NAFLD pathogenesis. In recent years, researchers have found that many gene polymorphisms are associated with NAFLD vulnerability through GWAS. Kawaguchi et al. were the first to find a significant association between SAMM50 gene polymorphisms and NAFLD susceptibility in the Japanese population via GWAS. Subsequently, the study was carried out by researchers from different countries, but the results were inconsistent. This inconsistency might be caused by factors like limited sample sizes, confounding factors, as well as clinical heterogeneity of NAFLD. Therefore, we collected the existing evidence and looked into the associations of SAMM50 gene SNPs and NAFLD vulnerability via meta-analysis, which could combine data from individual studies, examine and explain the heterogeneity, and increase the statistical power. In conclusion, the merged data suggested a significant correlation between rs2143571, rs3761472, and rs738491 polymorphisms of the SAMM50 gene and NAFLD vulnerability. Two large samples of GWAS were included in the original studies, with sufficient assurance to ensure the reliability of the study results. Of note, rigid methodological quality assessment, heterogeneity, and sensitivity analysis also ensured the robustness of the study results.Despite the specific pathological mechanism of NAFLD still needs to be explored. Nevertheless, research increasingly revealed that genetic predisposition plays a crucial intrinsic role in the occurrence and development of NAFLD.rs2143571, rs3761472, and rs738491 in the SAMM50 gene have a higher risk of NAFLD. Namely, it has a significantly increased risk for NAFLD. Furthermore, the outcomes of the meta-analysis were considered to be statistically robust and reliable according to the sensitivity analysis.In a word, we performed a meta-analysis of 8 case-control studies that satisfied the inclusion criteria. It should be noted that the current comprehensive analysis was more necessary and meaningful owing to the conclusions of qualified case-control studies are conflicting and contradictory. This work demonstrated that carriers of the A-allele of  It plays an important role in the formation of the mitochondrial crest, the assembly of the respiratory chain complex, and the maintenance of mitochondrial DNA stability. Studies have revealed that Sam50 reduction may lead to changes in mitochondrial shape and cristal morphology, and SAM50 consumption could affect the mitochondrial respiratory complexes. Ma et al. found a significant deletion of mitochondrial cristae in liver cells of patients with nonalcoholic steatohepatitis. In addition, changes in the normal structure of mitochondria could lead to insulin resistance, which is an important pathophysiological process of NAFLD.,35 All the above studies have indicated that the SAMM50 gene and its expression products play a crucial role in the development of NAFLD. According to the results of this meta-analysis, A allele of rs2143571, rs3761472, and rs738491 in the SAMM50 gene might be related to mitochondrial dysfunction and insulin resistance, leading to the NAFLD development.Sam50 is a kind of β-barrel protein distributed in the mitochondrial membrane and encoded by the SAMM50 gene.Up to now, this is the first synthetical study on the relationship between SAMM50 gene SNPs and NAFLD vulnerability. There were several strengths in this study. First, to gather a maximum amount of relevant literature, a comprehensive search strategy was adopted to retrieve eligible studies in both English and Chinese databases. Besides, the methodological quality of studies was evaluated via NOS, which allowed for the judgment of the potential risk of bias. According to the NOS, all eligible studies were of high methodological quality. Furthermore, sensitivity analyses were carried out in this study, which guaranteed the reliability of the findings.rs2143571, rs3761472, and rs738491 polymorphisms of the SAMM50 gene and NAFLD vulnerability.Nevertheless, there still existed several drawbacks that should be acknowledged. First, Although liver biopsy was regarded as the gold standard for NAFLD diagnosis, not all studies were included liver biopsy because pathology was expensive and invasive. Second, the original studies included in this study were mainly conducted in Asian and North American populations, and the results cannot reflect the situation of other populations. Third, We ignored the synergistic effect of polymorphisms at other sites of NAFLD because only 3 loci in the SAMM50 gene were studied in association with susceptibility to NAFLD. Thus, interactions between these loci and genes may result in concealing or amplifying the actual function of individual loci or genes. Leave aside these drawbacks, this study is the first to provide more accurate and powerful evidence on the association between rs2143571, rs3761472, and rs738491 polymorphisms of the SAMM50 gene are appreciably associated with an augmented risk of NAFLD vulnerability. In summary, this study will provide the latest evidence to support the susceptibility of SAMM50 gene polymorphisms and NAFLD, and provide strategies for the prevention and treatment of NAFLD. Concerning the limitations of this study, it is necessary to confirm the present findings with complementary studies with a larger sample size.This meta-analysis suggests that The authors would like to thank all studies and their participants involved in this investigation.Conceptualization: Ming Qiao.Supervision: Jian-hua Yang, Jun-ping Hu.Validation: Yi Zhu.Writing–original draft: Ming Qiao.Writing–review & editing: Jian-hua Yang, Jun-ping Hu.


Tenebrionoidea, with more than 30,000 described species and 30 currently recognized families, is a superfamily of difficult taxonomy. The aim of this work is to support the basal position of the Mordellidae among the beetle tenebrionoids. They have a low number of sperm cells per cysts, contrary to the more derived families of the group; moreover, their sperm are not distributed in two bundles at the opposite poles of the cysts, as occurs in the higher taxa, but their sperm flagella form a loop in the median region so that sperm nuclei are positioned close to the tail end. The sperm structure of two members of higher families, Oedemeridae and Tenebrionidae, are investigated to confirm the data mentioned above. The sperm looping, which also occurs in the closely related Ripiphoridae, could be the consequence of the growth asynchrony between the cyst size and the sperm length. The Mordellidae sperm are characterized, not by small mitochondrial derivatives and accessory bodies, but by a peculiar stiff and immotile thin flagellar posterior region provided with only accessory tubules.Mordellistena brevicauda and Hoshihananomia sp.), one species to Oedemeridae (Oedemera nobilis), and one species to Tenebrionidae (Accanthopus velikensis). It is confirmed that Mordellidae are characterized by the lowest number of spermatozoa per cyst (up to 64), a number shared with Ripiphoridae. In contrast, in the two other families, up to 512 spermatozoa per cyst are observed, the same number present, for example, in Tenebrionidae. Also, as in the other more derived families of tenebrionoids studied so far, during spermatogenesis in O. nobilis and A. velikensis, sperm nuclei are regularly distributed in two sets at opposite poles of the cysts. On the contrary, the Mordellidae species do not exhibit this peculiar process. However, during spermiogenesis, the bundles of sperm bend to form a loop in their median region, quite evident in the Hoshihananomia sp., characterized by long sperm. This process, which also occurs in Ripiphoridae, probably enables individuals to produce long sperm without an increase in testicular volume. The sperm looping could be a consequence of the asynchronous growth between cyst size and sperm length. The sperm ultrastructure of the Mordellidae species reveals that they can be differentiated from other Tenebrionoidea based on the shape and size of some sperm components, such as the accessory bodies and the mitochondrial derivatives. They also show an uncommon stiff and immotile posterior flagellar region provided with only accessory tubules. These results contribute to a better knowledge of the phylogenetic relationship of the basal families of the large group of Tenebrionoidea.The sperm ultrastructure of a few representative species of Tenebrionoidea was studied. Two species belong to the Mordellidae ( The Tenebrionoidea constitute one of the largest and most complex superfamilies of beetles ,2. A mol8) cells. In Tenebrionoidea, this number usually rises to 512 (29), but there are species where this number is only 64 (26), and there are also species where bundles contain 1024 (210) spermatozoa.The structure of tenebrionoid sperm is known from Baccetti et al. , Dias etPlanococcus citri (Pseudococcidae) [Kerria chinensis (Kerriidae) [Different morphological cellular mechanisms along the spermatogenic process in insects have shown a source of variability in sperm arrangements within cyst cells ,13. Accooccidae)  and Kerrrriidae) . In thisrriidae) , it was rriidae)  and McKerriidae) . The prerriidae)  and giveThe following species were studied in the present work:Mordellidae:Mordellistena (Mordellistena) brevicauda . Campiglia d’Orcia, Siena, Italy. 12 ex.Hoshihananomia sp. . Viçosa, MG, Brazil. 4 ex.Oedemeridae:Oedemera nobilis . Gallina, Siena, Italy. numerous ex.Tenebrionidae:Accanthopus velikensis . Monte Amiata, Siena, Italy. 2 ex.M. brevicauda, O. nobilis and A. velikensis were anesthetized with ether and dissected in 0.1 M phosphate buffer pH 7.2 with 3% of sucrose (PB) to remove the genital system. A drop of sperm, removed from the deferent duct and seminal vesicles, was spread over histological slides and photographed with a Leica DMRB light phase-contrast microscope. The length of spermatozoa was measured using image-J software. For the visualization of sperm nuclei, cells were spread on a histological slide, a drop of 1 ug/mL of the DNA specific dye Hoechst in 0.1 M PB was added, and the sample was finally covered with a glass coverslip. Fluorescence observations of the labelled samples were carried out with a Leica DMRB light microscope equipped with a UV light source, fluorescein, and UV filters and Zeiss AxioCam digital camera with dedicated imaging software.Males of Hoshihananomia sp., testes were dissected in PB and transferred to a 2% acetic-orcein solution. After 20 min, the follicles were placed on histological slides with a drop of acetic-orcein solution, dissociated using needles, and covered with coverslips. For testicular histology, testes were fixed in 2.5% glutaraldehyde solution in 0.1 M phosphate buffer, postfixed in 1% osmium tetroxide, dehydrated in alcohol solutions, and embedded in Historesin®. Semithin sections (0.5 µm thick) were stained with Giemsa for 15 min. To measure sperm length, the cells from the vas deferens were spread on histological slides and stained with Giemsa for 15 min. For nuclear size observations and measurements, some samples were stained for 20 min with 0.2 mg/mL DAPI, washed in distilled water, and mounted with 50% sucrose in PB.To observe the entire cysts in Mordellistena brevicauda were spread onto coverslips previously treated with poly-l-lysine. The coverslips were placed in 2.5% glutaraldehyde in PB for 30 min at 4 °C and then rinsed several times in PB. Specimens on glass coverslips were dehydrated in a graded series of ethanol and then processed by critical drying method in a Balzer’s CDP 030. The coverslips were sputtered with about 20 nm gold in a Balzer’s MED 010 sputtering device and finally observed in a SEM Phillips XL20 operating at 10 kV electron accelerating voltage.Mature spermatozoa taken from the seminal vesicles and deferent ducts of Adult males were dissected in PB to isolate the testes and deferent ducts. The material was fixed in 2.5% glutaraldehyde in PB overnight. After careful rinsing, the material was post-fixed in 1% osmium tetroxide for 2 h. After rinsing, the material was dehydrated with ethanol series (50% to 100%), then transferred to propylene oxide, and finally embedded in a mixture of Epon-Araldite resins. Some material was also treated with tannic acid, omitting osmium fixation. Ultrathin sections were obtained with a Reichert Ultracut ultramicrotome, routinely stained with uranyl acetate and lead citrate, and observed with a TEM Philips CM10 operating at 80 kV electron accelerating voltage.Hoshihananomia sp. were dissected, and the removed testes were processed following the conventional Transmission Electron Microscopy protocol. Ultrathin sections (~60 nm thick) were obtained with an ultramicrotome (Leica UC6). Then they were contrasted with solutions of 3% uranyl acetate and 0.2% lead citrate and after examined in a Transmission Electron Microscope (Tecnai G2-12—SpiritBiotwin FEI) operating at 120 kV at the Microscopy Center of the Federal University of Minas Gerais (CM-UFMG), Belo Horizonte, Minas Gerais, Brazil.Adult males of the species The male reproductive system consists of a pair of testes, each showing 4–5 ovoidal follicles, 480–500 µm long a,b. TestScanning electron microscopic preparations confirmed the sperm bundles bending at their half-length; the anterior nuclear region often appears disassembled, the middle regions are tightly twisted, and the posterior regions are thinner and stiff a–c. SperTwo antiparallel groups of packed sperm cells are visible in cross-sections cut at about half of the cyst length b, with eThe length of the sperm is 290–305 µm, of which 1.3 µm corresponds to a conical acrosome and 15 µm to the nucleus g. The acThe centriole consists of a complex of 9 microtubule doublets, devoid of dynein arms, and of a crown of 9 outer accessory tubules c. A crosHoshihananomia sp. is very similar to that of the previous species. Five follicles form each of the two testes; however, their follicles are elongated  sperm per cyst were observed, indicative of nine spermatogonial division cycles. On the other hand, in the Tenebrionidae Lagria villosa , 1024 (=210) sperm were counted per cyst [Ceracis cornifer , 256 sperm per cyst were observed [6) ([The sperm ultrastructure of the four species studied here not only confirms our previously published data (see ) but, inper cyst . Althougobserved , and theto 512 =2 sperm peto 512 =2 sperm peved [6)  in the proximal zone to the efferent duct (unpublished data). Such a feature is different from what was observed in Hoshihananomia sp., in which the long cysts are distributed along almost the entire central region of the follicles. These differences seem to validate an existing correlation between sperm and testis lengths and the organization of cysts within follicles.Testicular follicles in Gryllotalpa gryllotalpa [Zorotypus caudelli [Drosophila melanogaster and Bactrocera oleae [Mordellistena and Hoshihananomia [Mordellidae sperm (, this stllotalpa ,21, the caudelli , and in ra oleae . In the ra oleae , this stananomia , suggestananomia , the claO. nobilis and A. velikensis while in the two species of Mordellidae they are smaller and oval-shaped. The two accessory bodies, flanking the axoneme, are of primary importance to verify the relationship between the families of Tenebrionoidea. It has been already pointed out that the oval or elliptical shape of these structures in cross-section, is typical of various species of Tenebrionidae such as Tenebrio molitor or Tribolium castaneum [A. velikensis here studied. This shape, with few variations, is the commonest feature among the whole superfamily, and it was also found in Meloidae [O. nobilis, a member of the new family here studied has elliptical accessory bodies with a pointed apical side. On the contrary Mordellidae species have smaller accessory bodies with an almost triangular shape ([The sperm flagellar structure of insects is also characterized by the size and shape of the mitochondrial derivatives and the accessory bodies . All theastaneum ,9 and alMeloidae . O. nobiAs quoted in the Introduction, Levkaničová  and BocaThis study confirms that Mordellidae are closely related to Ripiphoridae and that both occupy a phylogenetically distinct position concerning other tenebrionoids. In mordellids, the number of sperm per cyst is low compared to the Oedemeridae and Tenebrionidae, also studied here. Furthermore, in these latter two families, the sperm of the same cyst are distributed in two sets arranged antiparallelly, as in the other families of Tenebrionoidea, except mordellids and ripiphorids. In these latter families, the long sperm cells exhibit the same orientation within each cyst, and the sperm bundle forms a loop approximately at half its length to be contained within a smaller cyst. Also, sperm are characterized by thin mitochondrial derivatives and accessory bodies and, in mordellids, by a long, rigid, immotile posterior flagellar region in which accessory tubules are embedded in dense intracellular material.


The prevalence of type 2 diabetes mellitus (T2DM) is growing worldwide. T2DM is often complicated by a range of psychological disorders that interfere with glycemic control and self-care. Previous studies have reported diabetes distress, depression, and anxiety among patients with T2DM; however; little is known about the burden of these comorbid mental disorders in primary care patients with T2DM treated in Egypt during the COVID-19 era. Participants were selected by convenient sampling from eight rural primary healthcare facilities from Ismailia in Egypt. Symptoms of diabetes distress, depression and anxiety were assessed by using the Arabic version of the 20-item Problem Areas in Diabetes (PAID), Patient Health Questionnaire 9, and Generalized Anxiety Disorder Scales, respectively. Multiple hierarchical logistic regression models were used to estimate the significant factors associated with diabetes distress, depression, and anxiety. A total of 403 individuals with T2DM were interviewed. The prevalence of severe diabetes distress was 13.4% (95% CI: 10.1–16.7), while prevalence of depressive and anxiety symptoms was 9.2% (95% CI: 6.4–12.0%), and 4.0% (95% CI: 2.1–5.9), respectively. In a series of hierarchical logistic regression models, significant predictors for diabetes distress were being married, illiterate, not-working, living with insufficient income, and having multi-comorbidities. Likewise, the significant predictors for depression and anxiety were elevated glycated hemoglobin level and the higher PAID total score, while having multi-comorbidities was a significant predictor for anxiety only. Diabetes distress was more prevalent than depressive and anxiety symptoms in this study population. Several sociodemographic and clinical characteristics were identified to be related with psychological problems among patients with T2DM, which necessitate a multidisciplinary team-based approach for optimal screening and management. Egypt has the 10th highest age-adjusted diabetes prevalence globally, with a comparative diabetes prevalence of 20.9% in people aged 20–79 years. T2DM is the most common form of DM, accounting for 90% of all cases of DM worldwide. T2DM can lead to premature death, a wide range of psychological disorders and decreased quality of life. Additionally, T2DM poses an economic burden on patients, families, and countries .The management of patients with T2DM is often complicated by a breadth of psychological disorders including diabetes distress (DD), depression, and anxiety which might negatively impact diabetic patients' quality of life and coping mechanisms with their disease , 6. The DD is the emotional burden associated with DM and its management over time , 10. A pDepressive symptoms are important indicators in individuals with T2DM. DD and depression are correlated and partly overlapping constructs but are not interchangeable . One metAnxiety is an emotion with important implications in patients with T2DM. Elevated anxiety symptoms were found in 40% of diabetic patients . The preThe American Diabetes Association notes that primary health care providers should consider evaluation for symptoms of DD, depression, and anxiety among PHC patients with T2DM using appropriate standardized and validated tools at their first visit, at periodic intervals, and when there is a change in illness, management, or life circumstance . AssessmThe present study is motivated by the paucity of current research on the prevalence of DD, depression, and anxiety symptoms within the T2DM population of Egypt considering the COVID-19 context. As such, the primary goal of our study is to assess the prevalence and associated factors of DD, depression, and anxiety symptoms. We also investigate the relationship between symptoms of DD, depression, and anxiety among PHC patients with T2DM during the COVID-19 pandemic in Egypt. We hypothesized that T2DM patients experienced high levels of DD, depression, and anxiety symptoms during the COVID-19 pandemic; symptoms of DD have a statistically significant positive relationship with depressive and anxiety symptoms; depressive symptoms have a statistically significant positive relationship with anxiety symptoms; and a certain set of demographic and clinical characteristics of patients with T2DM are related to symptoms of DD, depression, and anxiety.n = 25). Calculation was based on the least prevalence obtained from our pilot study (4% for anxiety symptoms), 2% margin of error, and 95% level of confidence. The calculated sample size was further increased by 10% to compensate for the non-response. A convenience sampling of 406 patients with T2DM who met the eligibility criteria were interviewed during the study period. Patients were eligible if they were 18 years or older, had been diagnosed with T2DM for at least 1 year, and gave a written informed consent to participate. Three patients were excluded who had gestational diabetes or were not able to give their consent due to a serious mental illness or cognitive impairment. So, 403 participants were included in our study.Using a cross-sectional design, this study was carried out in eight rural primary care facilities at the Ismailia governorate, Egypt during the COVID-19 pandemic (between September 2020 and June 2021). A sample size of 369 was calculated using Epi InfoTM StatCalc version 7.2.4.0 , given the data derived from a pilot study . All patients gave their written informed consent prior to their participation in this study.Data collection was performed using face-to-face interviews with selected patients. Questionnaire included questions about sociodemographic, lifestyle and clinical characteristics: age, gender, marital status, occupation, family income, duration of diabetes, treatment for diabetes, diabetes-related long-term complications , smoking, alcohol drinking, and physical activity. Patients were also asked about history of COVID-19 (confirmed or suspected). Furthermore, the PAID was used to assess DD , 44, 45,The PAID scale consisted of 20 items, with a total score ranged from 0 to 100. Each item scored on a 5-point Likert scale ranging from 0 to 4, where 0 = not a problem, and 4 = serious problem. PAID total score was calculated by summing all items and multiplying it by 1.25. A higher score indicating greater DD, with a score of ≥40 indicating severe emotional distress –39. The The PHQ-9 was used to evaluate the depressive symptoms where each item took a score from 0 to 3 . The total PHQ9 score was calculated as the sum of all items' scores, with a maximum score of 27. A total PHQ-9 score ≥10 showed a high sensitivity and specificity for major depression . The PHQThe GAD-7 was used to evaluate the anxiety symptoms with each item taking a score from 0 to 3 . The sum of all items' score comprised a total GAD-7 score ranging from 0 to 21. A total GAD-7 score of 10 or higher was satisfactory sensitive and specific for GAD . An ArabBody mass index (BMI) was calculated as the body weight (in kg) divided by the squared root of height (in meters), where patients were considered overweight if they had a BMI between 25 and 29.9, and obese if BMI ≥30. The world health organization has defined regular physical activity for adult people with chronic illness as engaging in at least 150 min or more of moderate-intensity aerobic activity per week; or at least 75 min or more of vigorous-intensity aerobic activity per week; or an equivalent combination of moderate- and vigorous-intensity activity weekly .The most recent HbA1c values  and lipid profile  were obtained from patients' medical records. Good glycemic control was identified if HbA1c values were <7% in adult, or <7.5% in adults older than 65 years .www.graphpad.com). Multiple hierarchical logistic regression models were used to identify the predictors of DD, depression, and anxiety symptoms among the studied patients. Independent variables were entered in the model as blocks. Three blocks were identified: the first block involved sociodemographic variables, second block included lifestyle and general health variables, while the third block included diabetes-related variables. Improvement in the predictive power of the consecutive models was identified by calculating the change in −2 log likelihood (−2LL) and was tested for statistical significance using the chi-square distribution (where the degree of freedom was the difference in the number of parameters in each model). Also, the change in the R-square and the predictive accuracy were reported for each model. Odds ratio (OR) and 95% confidence interval (CI) was reported for each independent variable in the models.All procedures of data management and analyses were performed with the Statistical Package for the Social Sciences (SPSS) for Windows, version 25.0 . A significance level was set at 0.05 for all statistical analyses. Categorical variables were described as frequencies and percentages (%), while numeric variables were summarized as mean and standard deviation. Associations between categorical variables were investigated for statistical significance with Chi-square test or Fischer's exact test as indicated. Graphs were created with GraphPad Prism , and 59.1% were female. Demographic, lifestyle, and health-related characteristics are presented in p < 0.001). Likewise, the PHQ-9 and the GAD-7 showed a significant, moderate, and positive correlation .According to the study findings, approximately one in seven, one in 10 and one in 25 primary care patients with T2DM in the rural area of Ismailia governorate in Egypt were suffering from symptoms of severe DD, major depression and anxiety, respectively. Several sociodemographic and clinical characteristics were identified to be associated with these findings at a different degree.The prevalence of DD in the current study was higher than what was reported in a meta-analysis by Perrin et al.  and in pJust <10% of our study sample had symptoms of major depression (PHQ-9 ≥10). Prevalence rates of comorbid depression in diabetes was variably reported with a range of 2–88% with a world-pooled prevalence of 28% . AccordiThe least reported comorbid psychiatric problem in patients with T2DM in this study was anxiety symptoms with only 4% of the study sample having a minimum score of 10 on the GAD questionnaire. Previous literature also showed higher rates of anxiety disorders among patients with T2DM ranging between 30 and 40%. However, these studies used different tools for anxiety assessment which makes direct comparison to our study findings difficult , 32, 37.Complex interactions between DM and social as well as cultural dynamics are likely to affect the way patients experience illness and health . Our stuRegarding age, bivariant analysis of this study suggested that symptoms of DD, depression, and anxiety occurred more frequently with increasing age. However, multivariant analysis did not find an association between age with either DD, depression or anxiety symptoms. Similarly, a Malaysian cross-sectional study on PHC patients with T2DM did not find a statistical association between age and prevalence of depressive and anxiety symptoms . on the The current study's bivariant analysis showed that illiteracy, insufficient family income, unemployment, and being divorced or widowed were associated with higher prevalence of DD, depression, and anxiety symptoms. Study findings from other countries also showed similar association although not all associations were statistically significant , 58. ConLifestyle factors like physical inactivity and obesity were associated with higher rates of diabetes associated mental health disorders in our study. This association was also confirmed in a longitudinal study that found that persistent depressive symptoms at 5 years were significantly associated with worse compliance with dietary and physical activity recommendations compared to patients with diabetes without depressive symptoms . MoreoveCOVID-19 pandemic exerted a tremendous pressure on both patients and healthcare providers with unknown long-term consequences. Although we did not find a significant association between history of COVID-19 infection and the prevalence of mental health disorders among diabetic patients in our study, an earlier longitudinal study in Australia found that COVID-19 lockdown restrictions had negative impact on patients with T2DM quality of life and physical activity levels without affecting generalized anxiety and depressive symptoms prevalence . TherefoIn the current study, diabetes duration, complications and treatment regimen were all associated with increased risks for comorbid DD, depression, and anxiety symptoms, which is consistent with previous studies , 55, 58.By using multiple logistic regression, we found that the only predictors for DD were social factors  and having multiple co-morbidities. Nevertheless, DD itself as well as HbA1c level were predictors for depressive and anxiety symptoms in patients with T2DM. The intercorrelation between social factors, health-related variables, DD, depression, and anxiety seem to go into a continuous cycle with complex interactions that necessitate a holistic patient-centered approach in order to break this cycle.Our study provided some of the early evidence on the burden of three important psychological disorders in people with T2DM who are managed in the PHC centers in the rural area of Ismailia in Egypt. However, this observational study is subjected to limitations. First, due to the cross-sectional nature of the study, it is not possible to identify causality between variables. Therefore, a further longitudinal study is needed to reveal the strength and direction of any potential association. Second, as we did not have baseline data on the burden on mental health disorders among patients with T2DM in our population, we could not assess the impact of COVID-19 pandemic on our study participants. Third, lack of randomization limited the ability to generalize the results. Fourth, the use of western methods to identify psychological disorders in non-western countries could be questionable. However, all the scales that we used  were validated for use in Arabic language. Nevertheless, developing culturally sensitive screening tools could help in better assessment of psychological disorders in non-English speaking patients with DM.Our study found DD is more prevalent than depressive and anxiety symptoms in adults with T2DM managed in the PHC facilities in the rural area of Ismailia in Egypt during the COVID-19 pandemic. DD and HbA1c level were associated with depressive and anxiety symptoms in this population of patients. Although psychosocial assessment is important for all patients with T2DM, our study findings suggest that PHC providers should pay closer attention to females, elderly, patients suffering from DM for a longer time, those with multiple comorbidities and/or chronic diabetes complications. As multiple sociodemographic and clinical factors were identified to be linked with psychological conditions in patients with T2DM, it is important to utilize multidisciplinary teams to achieve holistic patient-centered care. Further studies are necessary to better understand the long-term psychological consequences of the COVID-19 pandemic on patients with T2DM.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s on reasonable request.The studies involving human participants were reviewed and approved by Research Ethics Committee at the Faculty of Medicine, Suez Canal University, Ismailia, Egypt (Ref No. 4277/2020). The patients/participants provided their written informed consent to participate in this study.HS commenced the idea of this research. HS, AF, AJ, JS, and AS drafted this manuscript. SM collected the data of this research. AF and SE analyzed these data. SE, MM, JS, and SM critically reviewed this manuscript. SM and HS supervised this research. All authors designed this research and approved the final manuscript.AJ was employed by Hamad medical Corporation. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.