Survey on Perkinsus species in two economic mussels (Mytilus coruscus and M. galloprovincialis) along the coast of the East China Sea and the Yellow Sea.

Perkinsus, a parasitic pathogen of marine bivalves, is widely distributed among various mollusks in numerous countries. However, the prevalence and diversity of Perkinsus species in the two economically important mussels, Mytilus coruscus and M. galloprovincialis, in China remain unknown. The presence of the Perkinsus species was identified in the two mussels sampled along the coast of the East China Sea and the Yellow Sea, using both the alternative Ray's fluid thioglycolate medium (ARFTM) and conventional polymerase chain reaction (PCR). The ARFTM test indicated the presence of Perkinsus-like hypnospores in the two mussels. The diameter of the hypnospores in M. coruscus was significantly smaller than that in M. galloprovincialis. The prevalence of Perkinsus in M. galloprovincialis and M. coruscus ranged from 0 to 37.5% and 0 to 25%, respectively. The mean intensity of Perkinsus in M. galloprovincialis and M. coruscus ranged from 0 to 5.14 and 0 to 4.92, respectively. The PCR assay showed that the prevalence of Perkinsus spp. in M. galloprovincialis and M. coruscus was 0 to 25.0% and 0 to 12.5%, respectively. The homology analysis of the newly obtained internal transcribed spacer (ITS) sequences of Perkinsus revealed the highest identity of 100% with P. beihaiensis. The phylogenetic analysis indicated that the Perkinsus isolates from the two mussels were clustered with P. beihaiensis. The results of the molecular biology indicated that only P. beihaiensis was detected in the two mussels. The highest prevalence of P. beihaiensis was observed in Liaoning province (Dalian, 20.83%), followed by Shandong province, Zhejiang province and Fujian province. Consequently, it is recommended that surveillance should be conducted in Dalian, where the prevalence and mean intensity of P. beihaiensis in M. galloprovincialis are the highest.

Regulation of innate immunity in marine mussel Mytilus coruscus: MicroRNA Mc-novel_miR_196 targets McTLR-like1 molecule to inhibit inflammatory response and apoptosis.

Toll-like receptors (TLRs) are crucial players in immune recognition and regulation, with aberrant activation leading to autoimmune, chronic inflammatory, and infectious diseases. MicroRNAs (miRNAs) have been shown to regulate gene expression at transcriptional and post-transcriptional levels. While miRNA-mediated regulation of TLR signaling has been studied in mammals, the underlying mechanisms of TLR-miRNA interactions in molluscs remain unclear. In a previous study, one of the TLR genes potentially targeted by miRNAs was identified and named McTLR-like1. McTLR-like1 was later found to be targeted by miRNA Mc-novel_miR_196 through bioinformatic prediction. In this study, we aim to experimentally determine the interaction between McTLR-like1 and Mc-novel_miR_196, as well as their functional role in the innate immune response of molluscs. The results showed that the expression of Mc-novel_miR_196 was suppressed, while the expression of McTLR-like1 was enhanced in M. coruscus hemocytes treated with lipopolysaccharide (LPS). Moreover, in vitro assays demonstrated that Mc-novel_miR_196 directly targets the 5' UTR of McTLR-like1 and leads to the down-regulation of proinflammatory cytokines in hemocytes. In addition, co-transfection experiments confirmed that Mc-novel_miR_196 inhibits McTLR-like1 and inhibits the expression of proinflammatory cytokines. The Tunel assay also showed that Mc-novel_miR_196 inhibited apoptosis in hemocytes induced by LPS. Our findings suggest that microRNA Mc-novel_miR_196 acts as a regulator of innate immunity in M. coruscus by targeting McTLR-like1 and inhibiting inflammatory response and apoptosis. These results provide further insights into the complex molecular mechanisms underlying TLR signaling in molluscs.

Social support and subsequent cognitive frailty during a 1-year follow-up of older people: the mediating role of psychological distress.

BACKGROUND: Frailty and cognitive impairment are two common geriatric symptoms linking adverse health-related outcomes. However, cognitive frailty, a new definition defined by an international consensus group, has been shown to be a better predictor of increased disability, mortality, and other adverse health outcomes among older people than just frailty or cognitive impairment. This study estimated the prospective association between social support and subsequent cognitive frailty over 1 year follow-up, and whether psychological distress mediated the association. METHODS: The data was drawn from a prospective repeated-measures cohort study on a sample of participants aged 60 and over. A total of 2785 older people who participated in both of the baseline and 1-year follow-up survey were included for the analysis. Cognitive frailty was measured by the coexistence of physical frailty and cognitive impairment without dementia. Control variables included sex, age, education, marital status, economic status, smoking status, alcohol drinking status, chronic conditions, and functional disability. Path analyses with logistic function were performed to examine the direct effects of social support (predictors) on subsequent cognitive frailty (outcome) at 1-year follow-up and the mediating role of psychological distress (mediator) in this link. RESULTS: After adjusting for covariates and prior cognitive frailty status, social support was negatively associated with psychological distress (ß = - 0.098, 95% CI = - 0.137 to - 0.066, P < 0.001) and was negatively associated with the log-odds of cognitive frailty (ß = - 0.040, 95% CI = - 0.064 to - 0.016, P < 0.001). The magnitude of mediation effects from social support to cognitive frailty via psychological distress was a*b = - 0.009, and the ratio of a*b/(a*b + c') was 24.32%. CONCLUSIONS: Lower social support is associated with increased rates of subsequent cognitive frailty over 1-year follow-up, and this link is partially mediated through psychological distress, suggesting that assessing and intervening psychological distress and social support may have important implications for preventing cognitive frailty among older people.

Association between physical activity and falls among older adults in rural China: are there gender and age related differences?

BACKGROUND: The relationship between physical activity (PA) and falls among older adults is inconsistent, and little is known about the gender-specific association between falls and PA. Moreover, age may modify this relationship. This study aimed to test the association between PA and falls and to investigate the gender and age differences in the association among rural older adults. METHODS: This cross-sectional data were derived from the baseline survey of Shandong Rural Elderly Health Cohort (SREHC). In total, 3,242 rural older adults aged 60 years and above were included in the analysis. PA was measured by the International Physical Activity Questionnaire Short Form (IPAQ-S). PA levels were classified as low, moderate, elevated and high according to quartiles. Volume of moderate-to-vigorous physical activity (MVPA) was categorized into low, moderate, elevated, and high level based on global recommendations. Information on falls was determined from in-person interviews. Falling was defined to participants as ending up on the floor or ground because they were unable to stop themselves. Logistic regression analysis was employed to explore the association between falls and PA. RESULTS: Of 3,242 rural older adults, the incidence of falls was 13.1%. In older adults, high levels of PA [odds ratio (OR) = 0.65, 95% confidence interval (CI): 0.47-0.90] or MVPA (OR = 0.68, 95% CI: 0.50-0.94) were related to falls. Moderate (OR = 4.84, 95% CI: 1.68-13.94) or high (OR = 0.54, 95% CI: 0.30-0.99) levels of MVPA were associated with falls in older men. But elevated levels of PA were associated with falls (OR = 0.60, 95% CI: 0.42-0.87) in older women. Among older people younger than 75 years, elevated (OR = 0.54, 95% CI: 0.37-0.79) or high (OR = 0.68, 95% CI: 0.48-0.98) levels of PA were associated with falls. CONCLUSIONS: Among Chinese rural older adults, PA and MVPA are associated with falls, and there are gender and age differences. To prevent falls, measures need to account for individuals' gender and age to encourage rural older adults to participate more actively in PA. We will conduct longitudinal studies to clarify the causal relationship between PA and fall.

Family doctor contract services and health-related quality of life among patients with chronic diseases in rural China: what is the role of socioeconomic status?

PURPOSE: Few studies explored the relationship between the family doctor contract services (FDCS) and health-related quality of life (HRQOL) among patients with chronic diseases in rural China. This study aims to explore the relationship between the status of signing on FDCS and HRQOL among patients with chronic diseases and examine whether there are differences in the relationship between different socioeconomic status (SES). METHODS: A total of 1,210 respondents were included in this study. HRQOL was measured by EQ-5D-3L. The contracting status was divided into uncontracted and contracted. Tobit regression and Logistic regression were employed to explore the association between contracting status and HRQOL. The interaction terms were included to explore the differences in the association among different SES. RESULTS: Contracting with family doctors was associated with HRQOL (coefficient = 0.042; 95%CI 0.008 to 0.075). The association was different among different socioeconomic levels that the contracting status was only associated with HRQOL in sub-high-income (P < 0.01) and highly educated patients (P < 0.05). Compared with uncontracted patients, contracted patients reported higher ED-5D-3L utility value in the sub-high-income group (coefficient = 0.078; 95%CI 0.017 to 0.140) and high educational attainment (coefficient = 0.266; 95%CI 0.119 to 0.413). CONCLUSIONS: This study found a significant association between FDCS and HRQOL among chronic patients in rural Shandong, China. This relationship varied by income levels and educational attainment. The government should take efforts to formulate a variety of measures to encourage chronic patients to contract with family doctors, with special attention to people with low SES.

Changes in psychological distress before and during the COVID-19 pandemic among older adults: the contribution of frailty transitions and multimorbidity.

AIM: To investigate changes in psychological distress in community-dwelling older adults before and during the coronavirus disease 2019 (COVID-19) pandemic and the contribution of frailty transitions and multimorbidity in predicting the psychological distress. METHODS: Prospective repeated-measures cohort study on a sample of participants aged 60 and over. A total of 2, 785 respondents at the baseline (May 2019) were followed during the COVID-19 (August 2020). The changes in psychological distress before and during the COVID-19 were assessed using generalised estimation equations with adjusting for sex, age, education, economic status, marital status, tea drinking status, smoking status, alcohol drinking status, sedentary time, sleep quality and activities of daily living. RESULTS: The psychological distress of older people has significantly increased in August 2020 compared with May 2019. Both older adults who remained frail and transitioned into frail state reported more psychological distress during the COVID-19. Similarly, both pre-existing multimorbidity and emerging multimorbidity groups were associated with more psychological distress. The group of frailty progression who reported new emerging multimorbidity showed more increase in psychological distress in comparison with those who remained in the non-frail state who reported no multimorbidity. CONCLUSION: Psychological distress has increased among the community-dwelling older adults during the COVID-19 pandemic, and sustained and progressive frail states as well as multimorbidity were all associated with a greater increase of psychological distress. These findings suggest that future public health measures should take into account the increased psychological distress among older people during the COVID-19 pandemic, and the assessment of frailty and multimorbidity might help in warning of psychological distress.

Body mass index and mild cognitive impairment among rural older adults in China: the moderating roles of gender and age.

BACKGROUND: Evidence concerning the association between body mass index (BMI) and cognitive function among older people is inconsistent. This study aimed to investigate gender and age as moderators in association between BMI and mild cognitive impairment (MCI) among rural older adults. METHODS: Data were derived from the 2019 Health Service for Rural Elderly Families Survey in Shandong, China. In total, 3242 people aged 60 years and above were included in the analysis. Multilevel mixed-effects logistic regression was used to examine the moderating roles of gender and age, then further to explore the relationship between BMI and MCI. RESULTS: There were 601 (18.5%) participants with MCI. Compared with normal BMI group, low BMI group had a higher risk of MCI among older people [adjusted odds ratio (aOR) = 2.08, 95% confidence interval (CI): 1.26-3.44], women (aOR = 2.06, 95% CI: 1.35-3.12), or the older elderly aged ≥75 years old (aOR = 3.20, 95% CI: 1.34-7.45). This effect remained statistically significant among older women (aOR = 3.38, 95% CI: 1.69-6.73). Among older men, elevated BMI group had a higher risk of MCI (aOR = 2.32, 95% CI: 1.17-4.61) than normal BMI group. CONCLUSIONS: Gender and age moderated the association between BMI and MCI among Chinese rural older adults. Older women with low BMI were more likely to have MCI, but older men with elevated BMI were more likely to have MCI. These findings suggest rural community managers strengthen the health management by grouping the weight of older people to prevent the risk of dementia.

Intergenerational support and subjective wellbeing among oldest-old in China: the moderating role of economic status.

BACKGROUNDS: The oldest-old population is increasing sharply in China, and intergenerational support has been their primary source of caregiving. Although intergenerational support has been found to be associated with wellbeing of older people in previous study, most analysis were from the perspective of children's characteristics and exchange patterns. This study aims to investigate the impact of different types of intergenerational support on subjective wellbeing among Chinese oldest-old and the variation across groups of different economic status, based on their five-tier of needs (physiological needs, safety needs, love/belonging needs, esteem needs, and self-actualization needs). METHODS: We included older adults aged ≥ 80 years from the 2018 Chinese longitudinal Healthy Longevity Survey (CLHLS). We assessed older people's subjective wellbeing by their life satisfaction and psychological health. We evaluated four types of intergenerational support: parents provide financial support, receive financial, instrumental and emotional support. We applied binary logistic regression analysis to analyze the association between different intergenerational support and older people's subjective wellbeing and the moderating effect of self-rated economic status on this relationship. RESULTS: A total of 8.794 participants were included, with a mean age of 91,46 years (standard deviation:7.60). Older adults who provide financial support (OR: 1.37, 95% CI: 1.01, 1.85) and receive emotional support (OR: 1.99, 95% CI: 1.40, 2.83) report better subjective wellbeing. However, receiving instrumental support depressed psychological health (OR: 0.67, 95% CI: 0.56, 0.79) while improved life satisfaction (OR: 1.42, 95% CI: 1.04, 1.55). Receiving emotional support promoted parents' psychological health among all combinations of support, and receiving all the three types together raised their subjective wellbeing most. CONCLUSIONS: Our study recognizes that higher level of subjective wellbeing for oldest-old is related to providing financial support, receiving emotional and certain instrumental support. In addition, higher economic status can moderate these associations.

Do older people with cardiovascular-metabolic multimorbidity prefer to sign contracts for family doctor services? Evidence from a cross-sectional study in rural Shandong, China.

BACKGROUND: Family doctor policy is an important part of deepening healthcare reform in China. The study aimed to explore the association between cardiovascular-metabolic multimorbidity and the status of signing a contract for family doctor services among the older people in rural Shandong, China. METHODS: A cross-sectional study was conducted in 3 cities of Shandong province, China. A total of 1395 rural residents over 60 years of age were included in this study using a multistage stratified random sampling method. Covariates included demographic and socioeconomic characteristics, health-related characteristics, health service utilization, and awareness of family doctor contract services. The univariate and multivariate regression logistic analysis was used to analyze the data. RESULTS: There were 28.2% of the rural older people contracted for the family doctor contract services. The contract rate of seniors with cardiovascular-metabolic multimorbidity was statistically higher than those without cardiovascular-metabolic multimorbidity (OR = 1.67, 95%CI, 1.21-2.32) after controlling for confounding factors. In addition, occupation, physical activities, self-rated health status, distance from the village clinic, the awareness of family doctor contract services were found to be associated with the signing behavior among the rural older adults. CONCLUSION: This study demonstrated that the rural older people with cardiovascular-metabolic multimorbidity had a higher family doctor contract rate than those without cardiovascular-metabolic multimorbidity, and there was a gap between the current signing rate and the policy goal. To increase the rate of signing for family doctor contract services, the government should take joint efforts to expand the publicity and coverage, and give priority to meeting the healthcare demands of rural older adults with cardiovascular-metabolic multimorbidity.

Family characteristics associated with rural households' willingness to renew the family doctor contract services: a cross-sectional study in Shandong, China.

BACKGROUND: In China, some previous studies have investigated the signing rate and willingness of residents to sign the family doctor contract services (FDCS). Few studies have explored residents' willingness to renew the FDCS. This study is designed to understand the family characteristics difference towards rural households' willingness of maintaining the FDCS. METHODS: A total of 823 rural households were included in the analysis. A descriptive analysis was conducted to describe the sample characteristics. The binary logistic regression model was used to explore the family characteristics that influence the renewal willingness for FDCS among rural households in Shandong province, China. RESULTS: Our study found that about 95.5% rural households had willingness to maintain the FDCS in Shandong, China. Those households with catastrophic health expenditures (CHE) (OR = 0.328, 95%CI = 0.153-0.703), with highest level of education at graduate or above (OR = 0.303, 95%CI = 0.123-0.747) were less willing to maintain the FDCS. Those whose households have more than half of the labor force (OR = 0.403, 95%CI = 0.173-0.941) and those households living in economically higher condition were less willing to maintain the FDCS. CONCLUSIONS: This study demonstrates a significant association between family characteristics (CHE, highest education in households, proportion of the household labor force) and willingness to maintain FDCS among rural households in Shandong, China. Targeted policies should be made for rural residents of identified at-risk families.

Willingness of patients with chronic disease in rural China to contract with family doctors: implication for targeting characteristics.

BACKGROUND: Rural residents with chronic conditions have a stronger need for health services, which should make using family doctor contract services a priority. This study aimed to evaluate the rate of willingness among rural residents with chronic conditions to contract with family doctors and examine its determinants. METHODS: A cross-sectional study was conducted from May, 2018 to June, 2018 in Shandong Province in China. A total of 769 rural unsigned residents with chronic conditions were included in the analysis. Using the Andersen model as the theoretical framework, logistic regression models were chosen to analyse the factors associated with willingness to contract with family doctors. RESULTS: This study found that the rate of willingness to contract with family doctors among chronic patients in rural Shandong was 46.7%. A higher willingness was observed in those living a further distance from the village clinic (more than 600 m: OR = 1.85, 95%CI =1.17-2.93), having received publicity for family doctor contract services (OR = 1.71, 95% CI = 1.06-2.76), reporting need for utilizing a chronic disease management program (OR = 3.36, 95% CI = 2.20-5.23), and reporting need for higher medical insurance reimbursement (OR = 1.91, 95% CI = 1.28-2.83). CONCLUSIONS: The prevalence of contract willingness was relatively low among unsigned rural residents with chronic conditions in rural Shandong, China. The need factors were powerful factors affecting their willingness to contract with family doctors. The government should therefore strengthen targeted publicity and education to rural residents with chronic conditions and provide targeted healthcare services, such as chronic disease management programs and medical services with higher reimbursement rates, to promote their willingness to contract with family doctors.

What Role Does Activity Engagement Play in the Association between Cognitive Frailty and Falls among Older Adults? Evidence from Rural Shandong, China.

INTRODUCTION: Previous studies have demonstrated the relationship between cognitive frailty and falls among older adults. Activity engagement (AE) is known to be related to falls in older adults but the subject has been limited to empirical study. This study aimed to explore the mediating role of AE between cognitive frailty and falls among older adults in rural Shandong, China. METHODS: A total of 3,242 rural seniors (age ≥60 years; 63.6% women) were included in this cross-sectional study. Regression and bootstrap analyses were performed to explore the mediating role of AE between cognitive frailty and falls. RESULTS: The prevalence of falls was 13.1% and the prevalence of cognitive frailty was 6.6% among the participants. AE mediated the association between cognitive frailty and falls (95% CI 0.077-0.223). However, the direct effect was no longer significant after being adjusted for AE (95% CI -0.037 to 0.684; p = 0.078). CONCLUSION: Cognitive frailty was found to be associated with falls among rural older adults, and AE mediated this association. More attention should be paid to promote AE among Chinese rural older adults with cognitive frailty.

Analysing the preferences for family doctor contract services in rural China: a study using a discrete choice experiment.

BACKGROUND: Preliminary evaluations have found that family doctor contract services (FDCSs) have significantly controlled medical expenses, better managed chronic diseases, and increased patient satisfaction and service compliance. In 2016, China proposed the establishment of a family doctor system to carry out contract services, but studies have found the uptake and utilization of these services to be limited. This study aimed to investigate rural residents' preferences for FDCSs from the perspective of the Chinese public. METHODS: A discrete choice experiment (DCE) was performed to elicit the preferences for FDCSs among rural residents in China. Attributes and levels were established based on a literature review and qualitative methods. Five attributes, i.e., cost, medicine availability, the reimbursement rate, family doctor competence, and family doctor attitude, were evaluated using a mixed logit model. RESULTS: A total of 609 residents were included in the main DCE analysis. The respondents valued the high competence (coefficient 2.44, [SE 0.13]) and the good attitude (coefficient 1.42, [SE 0.09]) of family doctors the most. Cost was negatively valued (coefficient - 0.01, [SE 0.01]), as expected. Preference heterogeneity analysis was conducted after adjusting the interaction terms, and we found that rural residents with higher educational attainment prefer a good attitude more than their counterparts with lower educational attainment. The estimated willingness to pay (WTP) for "high" relative to "low" competence was 441.13 RMB/year, and the WTP for a provider with a "good" attitude relative to a "poor" attitude was 255.77 RMB/year. CONCLUSION: The present study suggests that strengthening and improving the quality of primary health care, including the competence and attitudes of family doctors, should be prioritized to increase the uptake of FDCSs. The contract service package, including the annual cost, the insurance reimbursement rate and individualized services, should be redesigned to be congruent with residents having different health statuses and their stated preferences.

Cloning and characterization of the LEF/TCF gene family in grass carp (Ctenopharyngodon idella) and their expression profiles in response to grass carp reovirus infection.

T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) proteins from the High Mobility Group (HMG) box family act as the main downstream effectors of the Wnt signaling pathway. HMGB proteins play multifaceted roles in the immune system of mammals. To clarify the immunological characteristics of LEF/TCF genes in grass carp (Ctenopharyngodon idella), five LEF/TCF genes (TCF7, LEF1, TCF7L1A, TCF7L1B, and TCF7L2) were identified and characterized. All five LEF/TCF proteins contained two characteristic domains: a HMG-BOX domain and a CTNNB1_binding region. Phylogenetic tree analysis revealed that the LEF/TCF proteins were represented different lineages. These results of subcellular localization showed that four of the LEF/TCF genes were localized exclusively within the nucleus, while TCF7L2 was localized in the cytoplasm and nucleus. The mRNA expression profiles of these LEF/TCF family genes differed across different tissues. The mRNA expression levels of TCF7, TCF7L1A, and TCF7L2 changed significantly in liver after grass carp reovirus (GCRV) challenge; TCF7 and TCF7L1A responded early while TCF7L2 responded late. This suggests that these genes may participate in GCRV-related immune responses. Moreover, TCF7 promoted Bcl6 transcription in response to the GCRV challenge. These findings further our understanding of the function of LEF/TCF genes in teleosts.

Molecular characterization, tissue distribution and functional analysis of galectin 1-like 2 in grass carp (Ctenopharyngodon idella).

Galectins, as an evolutionary conserved group of lectin superfamily, has the functions of pathogen recognition, anti-bacteria and anti-virus. In this study, a 405 bp cDNA sequence of galectin 1-like 2 (CiGal1-L2) was obtained from grass carp (Ctenopharyngodon idella), which encoded 134 amino acids with a predicted molecular mass of 15.143 kDa and an isoelectric point of 5.33. The sugar binding motifs (H-N-R, V-N and W--E-R) were detected in carbohydrate-binding domain (CRD). The amino acid sequence similarity showed that CiGal1-L2 was 40.30-42.54% and 66.42-81.20% similarity to mammalian and fish counterparts, respectively. The phylogenetic tree showed that CiGal1-L2 was clustered with fish galectin-1s and closely related to Cyprinus carpio. Real-time quantitative PCR (RT-qPCR) analysis revealed that CiGal1-L2 was widely expressed in all tested tissues. In addition, the expression of CiGal1-L2 was differentially up-regulated challenged with grass carp reovirus (GCRV), lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C). The fluorescence of CiGal1-L2-GFP was distributed in the cytoplasm and nucleus of HEK 293T cells and showed a trend of nuclear translocation after LPS and poly I:C treatment. Finally, the recombinant CiGal1-L2 (rCiGal1-L2) protein showed strong binding ability to LPS. In conclusion, the results provided further insight into the immune roles of galectin-1 in teleost.

Identification and molecular characterization of peroxiredoxin 2 in grass carp (Ctenopharyngodon idella).

Peroxiredoxin (Prx), also named thioredoxin peroxidase (TPx), is a selenium independent antioxidant enzyme that can protect organisms from oxidative damage caused by reactive oxygen species (ROS) and is important for immune responses. In this study, the molecular cloning and characterization of a Prx2 homologue (CiPrx2) were described from grass carp (Ctenopharyngodon idella). The full-length cDNA of CiPrx2 was 1163 bp containing 5'-untranslated region (UTR) of 52 bp, a 3'-UTR of 517 bp with the putative polyadenylation consensus signal (AATAAA), an open reading frame (ORF) of 594 bp encoding polypeptides of 197 amino acids with a predicted molecular mass of 21.84 kDa and theoretical isoelectric point of 5.93. The analysis results of multiple sequence alignment and phylogenetic tree confirmed that CiPrx2 belong to the typical 2-Cys Prx subfamily. The CiPrx2 mRNA was ubiquitously expressed in all tested tissues. The temporal expression of CiPrx2 were differentially induced infected with grass carp reovirus (GCRV), polyinosinic:polycytidylic acid (poly I:C) and lipopolysaccharide (LPS) in liver and spleen. Subcellular localization of CiPrx2-GFP fusion proteins were only distributed in the cytoplasm. The purified recombinant CiPrx2 possessed an apparent antioxidant activity and could protect DNA against oxidative damage. Finally, CiPrx2 proteins could obviously inhibit H2O2 and heavy metal toxicity. However, further researches are needed to better understand the regulation of CiPrx2 under oxidative stresses.

Molecular characterization and functional activity of Prx1 in grass carp (Ctenopharyngodon idella).

Peroxiredoxin (Prx) family are known as an important antioxidant enzyme as the first line of defense against oxidative damage, and also involved in immune responses following viral and bacterial infection. Here, a full-length Prx1 cDNA sequence (CiPrx1) was cloned from grass carp (Ctenopharyngodon idella), which was 1029 bp, including a 5'-terminal untranslated region (UTR) of 121 bp, a 3'-UTR of 272 bp, an open reading frame of 600 bp encoding 199 amino acids with molecular weight of 22.21 kDa and isoelectric point of 6.30. CiPrx1 shares 80.8-99% protein sequence similarity with Prx1 of other fishes. The conserved peroxidase catalytic center "FYPLDFTFVCPTEI" and "GEVCPA" were observed in the sequence of CiPrx1; this indicated that it was a member of 2-Cys Prx. Subcellular localization of CiPrx1 was only strongly distributed in the cytoplasm. Quantitative real-time PCR (RT-qPCR) assays revealed that CiPrx1 mRNA was ubiquitously detected in all tested tissues, and the expression was comparatively high in liver, gill and spleen. Further, the expression of CiPrx1 can be induced by grass carp reovirus (GCRV), lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (Poly I:C) infection in the different tissues. Moreover, the recombinant CiPrx1 (rCiPrx1) protein was found a potential antioxidant enzyme, that could inhibit DNA damage from oxidants. Altogether, our results imply that CiPrx1 is associated with defending against virus and bacteria pathogens and oxidants in grass carp.

Investigating the Role of BATF3 in Grass Carp (Ctenopharyngodon idella) Immune Modulation: A Fundamental Functional Analysis.

Basic leucine zipper transcription factor ATF-like (BATF)-3, belonging to activator protein 1 (AP-1) superfamily transcription factors, is essential for homeostatic development of CD8α⁺ classical dendritic cells activating CD8 T-cell responses to intracellular pathogens. In this study, the characteristics and cDNA cloning of the CiBATF3 molecule were described in grass carp (Ctenopharyngodon idella). CiBATF3 had abundant expression in immune-related organizations, including liver, spleen and gill, and grass carp reovirus (GCRV) infection had significantly changed its expression level. After Ctenopharyngodon idella kidney (CIK) cells were challenged with pathogen-associated molecular patterns (PAMPs), polyinosinic:polycytidylic acid (poly(I:C)) stimulation induced higher mRNA levels of CiBATF3 than that of lipopolysaccharide (LPS). Subcellular localization showed that CiBATF3-GFP was entirely distributed throughout cells and nuclear translocation of CiBATF3 was found after poly(I:C) treatment. Additionally, the interaction between CiBATF3 and interleukin 10 (IL-10) was proven by bimolecular fluorescence complementation (BiFC) system. The small interfering RNA (siRNA)-mediated CiBATF3 silencing showed that the mRNA of CiBATF3 and its downstream genes were down-regulated in vitro and in vivo. CiBATF3 played a negative regulatory role in the transcriptional activities of AP-1 and NF-κB reporter gene. In summary, the results may provide valuable information on fundamental functional mechanisms of CiBATF3.

Associations Between Gene Polymorphisms and Psychological Stress in the Guangxi Minority Region of China.

To investigate whether there is an association between gene polymorphisms, genetic and environmental interactions, and psychological stress reactivity in Chinese subjects living in the Guangxi minority region. This cross-sectional study enrolled subjects older than 18 years, living in Nandan county, Guangxi minority region, China for at least 1 year. All participants were healthy, without any mental diseases, and were able to communicate. Eligible participants were randomly selected. The Life Event Scale Questionnaire, Simplified Coping Style Questionnaire, and Social Support Rating Scale were used to measure the physiological stress, coping style, and social support, respectively, in individuals. A total of 600 participants were recruited. A decreased risk of psychological stress was only found in TT of NPSR1 (rs324981): A allele carriers vs. TT genotype (OR 1.64, 95% CI 1.11, 2.42), and AT genotype vs. TT genotype (OR 1.76, 95% CI 1.17, 2.65). The overall coping style was positively associated with psychological stress, and no significant interactions between genetics and environment were found. We found that the NPSR1 (rs324981) T/T genotype decreased the risk of psychological stress, while the overall coping style was a risk factor for psychological stress. However, there was no interactive effects of genes and environment on psychological stress. Our findings will improve understanding of the biological basis underlying psychological stress if the results can be replicated in further research.

Oridonin alleviates inflammation and endoplasmic reticulum stress in pediatric pneumonia via regulating the SIRT1-mediated Wnt/ß-catenin signaling pathway.

BACKGROUND: Pediatric pneumonia is a prevalent and significant health concern worldwide, with elevated morbidity and mortality rates among affected children. This study was designed to elucidate the therapeutic impact of Oridonin (Ori) on pediatric pneumonia and unravel the underlying mechanisms involved. METHODS: A pediatric infantile pneumonia model was established in mice through intratracheal administration of LPS. Additionally, a cell damage model was created in WI-38 cells by administering LPS. Protein levels were assessed via western blotting, and cell viability was measured with CCK-8. Inflammatory cytokines were quantified through ELISA, and specific assays were employed to evaluate oxidative stress markers. Flow cytometry was utilized to assess cell apoptosis. RESULTS: Ori alleviated lung inflammation, oxidative stress, apoptosis, and endoplasmic reticulum stress (ERS) in LPS-induced pneumonia mice. In addition, Ori increased the viability of LPS-induced pneumonia cells but decreased cell apoptosis. Furthermore, Ori reduced oxidative stress, inflammation, and ERS in LPS-induced pneumonia cells by enhancing SIRT1 to activate the Wnt/ß-catenin pathway. CONCLUSION: This study suggested that Ori inhibited pediatric pneumonia by dampening the inflammatory response, oxidative stress, cell apoptosis, and ERS via the SIRT1/Wnt/ß-catenin pathway.