Combined healthy lifestyle behaviours and incident dementia: A systematic review and dose-response meta-analysis of cohort studies.

BACKGROUND: The associations of combined healthy lifestyle behaviours and incident dementia have not been systematically reviewed and the dose-response relationship was uncertain. OBJECTIVES: To evaluate the associations of combined healthy lifestyle behaviours with incident dementia and other cognitive outcomes, assess the dose-response relationship between the number of lifestyle behaviours and incident dementia, and summarise the adherence to healthy lifestyle behaviours. DESIGN: Systematic review and meta-analysis. METHODS: PubMed, EMBASE, Web of Science and PsycINFO were searched from inception to 20 Jan 2024. Cohort studies reporting associations of combined healthy lifestyle behaviours with incident dementia or other cognitive outcomes were included. We used the random-effects meta-analysis to pool the risk estimates and the robust error meta-regression method to examine the dose-response relationship. The methodological quality was assessed using the Newcastle-Ottawa Scale. RESULTS: A total of 22 articles including 25 cohort studies mostly from high-income economics were included, with all assessed as high methodological quality. Adherence to a healthy lifestyle was associated with a decreased risk of incident dementia, either per healthy lifestyle behaviour increase (pooled hazard ratio 0.89, 95 % confidence interval 0.85-0.94) or the highest level versus the lowest level (pooled hazard ratio 0.61, 95 % confidence interval 0.49-0.76). An inverse, linear dose-response relationship (Pnon-linear = 0.845) between the number of healthy lifestyle behaviours and incident dementia was observed, with an 11 % risk reduction for each healthy behaviour increase. A relatively limited number of included studies indicated that adherence to a healthy lifestyle combination could yield benefits for cognitive decline, global cognition, memory and executive function. In addition, the adherence rates typically decreased as the number of healthy lifestyle behaviours increased. CONCLUSIONS: Adherence to a healthy lifestyle was associated with a lower risk of incident dementia and other cognitive outcomes. It is important to find a subtle balance between the benefits and adherence. Further large cohort studies for combined lifestyle behaviours with specific cognitive outcomes and dose-response relationships are required, especially based on middle- and low-income populations. REGISTRATION: The study was registered in PROSPERO (CRD42023418509). TWEETABLE ABSTRACT: Engaging in a greater number of healthy lifestyle behaviours yields increased benefits in preventing dementia, albeit with lower adherence rates as a trade-off. Finding a delicate balance between the benefits and adherence is crucial.

Effects of Otago exercise program on physical function in older adults: A systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Maintaining physical function is critical for older adults to achieve healthy aging. The Otago exercise program (OEP) has been widely used to prevent falls for older adults. However, the effects of OEP on physical function remain controversial and the possible effects modifiers have not been assessed. OBJECTIVE: To evaluate the effects of OEP on physical function in older adults and to explore potential moderators underlying the effects of OEP. METHODS: We searched five electronic databases and relevant systematic reviews to identify studies. We included randomized controlled trials (RCTs) evaluating the effects of OEP as a single intervention on physical function among older adults aged 65 and over. Meta-analysis was performed using the random-effects model. Standardized mean differences (SMD) for physical function changes, pertinent to balance, strength, and mobility, were outcome measures. Subgroup analyses on exercise protocol and participants' characteristics were performed. RESULTS: Thirteen RCTs consisting of 2402 participants were included in this systematic review and meta-analysis. Results indicated a significant effect of OEP on balance (SMD = 0.59, 95 % CI: 0.22∼0.96), lower body strength (SMD = 0.93, 95 % CI: 0.31∼1.55), and mobility (SMD = -0.59, 95 % CI: -0.95∼-0.22) against control groups. No significant OEP effects were found on upper body strength (MD = 1.48, 95 % CI: -0.58∼3.55). Subgroup analysis revealed that the video-supported delivery mode was more effective for improving balance (P = 0.04) and mobility (P = 0.02) than the face-to-face mode. Session durations over 30 min was more effective on lower body strength (P < 0.001) and mobility (P < 0.001) than those 1-30 min. Program period of 13-26 weeks was more effective on mobility (P = 0.02) than those of 4-12 weeks. However, the effects of OEP on physical function were not associated with age groups, and baseline falling risks. CONCLUSION: The OEP could improve physical function including balance, lower body strength, and mobility in older adults. Implementing the OEP in video-supported, more than 30 min per session and 4-12 weeks may be the most appropriate and effective exercise protocol for improving physical function among older adults. More RCTs with rigorous design and larger scale are needed to further assess the effectiveness of diverse OEP protocols and quantify the dose-effect relationship.

Unveiling the link between physical activity levels and dementia risk: Insights from the UK Biobank study.

BACKGROUND: There is limited information on the mixture effect and weights of light physical activity (LPA), moderate physical activity (MPA), and vigorous physical activity (VPA) on dementia risk. METHODS: A prospective cohort study was conducted based on the UK Biobank dataset. We included participants aged at least 45 years old without dementia at baseline between 2006-2010. The weighted quantile sum regression was used to explore the mixture effect and weights of three types of physical activity on dementia risk. RESULTS: This study includes 354,123 participants, with a mean baseline age of 58.0-year-old and 52.4 % of female participants. During a median follow-up time of 12.5 years, 5,136 cases of dementia were observed. The mixture effect of LPA, MPA, and VPA on dementia was statistically significant (ß: -0.0924, 95 % Confidence Interval (CI): (-0.1402, -0.0446), P < 0.001), with VPA (weight: 0.7922) contributing most to a lower dementia risk, followed by MPA (0.1939). For Alzheimer's disease, MPA contributed the most (0.8555); for vascular dementia, VPA contributed the most (0.6271). CONCLUSION: For Alzheimer's disease, MPA was identified as the most influential factor, while VPA stood out as the most impactful for vascular dementia.

Whole genome sequencing identified genomic diversity and candidated genes associated with economic traits in Northeasern Merino in China.

Introduction: Northeast Merino (NMS) is a breed developed in Northeast China during the 1960s for wool and meat production. It exhibits excellent traits such as high wool yield, superior meat quality, rapid growth rate, robust disease resistance, and adaptability to cold climates. However, no studies have used whole-genome sequencing data to investigate the superior traits of NMS. Methods: In this study, we investigated the population structure, genetic diversity, and selection signals of NMS using whole-genome sequencing data from 20 individuals. Two methods (integrated haplotype score and composite likelihood ratio) were used for selection signal analysis, and the Fixation Index was used to explore the selection signals of NMS and the other two breeds, Mongolian sheep and South African meat Merino. Results: The results showed that NMS had low inbreeding levels, high genomic diversity, and a pedigree of both Merino breeds and Chinese local breeds. A total length of 14.09 Mb genomic region containing 287 genes was detected using the two methods. Further exploration of the functions of these genes revealed that they are mainly concentrated in wool production performance (IRF2BP2, MAP3K7, and WNT3), meat production performance (NDUFA9, SETBP1, ZBTB38, and FTO), cold resistance (DNAJC13, LPGAT1, and PRDM16), and immune response (PRDM2, GALNT8, and HCAR2). The selection signals of NMS and the other two breeds annotated 87 and 23 genes, respectively. These genes were also mainly focused on wool and meat production performance. Conclusion: These results provide a basis for further breeding improvement, comprehensive use of this breed, and a reference for research on other breeds.

Concurrent Encoding of Sequence Predictability and Event-Evoked Prediction Error in Unfolding Auditory Patterns.

Human listeners possess an innate capacity to discern patterns within rapidly unfolding sensory input. Core questions, guiding ongoing research, focus on the mechanisms through which these representations are acquired and whether the brain prioritizes or suppresses predictable sensory signals. Previous work, using fast auditory sequences (tone-pips presented at a rate of 20 Hz), revealed sustained response effects that appear to track the dynamic predictability of the sequence. Here, we extend the investigation to slower sequences (4 Hz), permitting the isolation of responses to individual tones. Stimuli were 50 ms tone-pips, ordered into random (RND) and regular (REG; a repeating pattern of 10 frequencies) sequences; Two timing profiles were created: in "fast" sequences, tone-pips were presented in direct succession (20 Hz); in "slow" sequences, tone-pips were separated by a 200 ms silent gap (4 Hz). Naive participants (N = 22; both sexes) passively listened to these sequences, while brain responses were recorded using magnetoencephalography (MEG). Results unveiled a heightened magnitude of sustained brain responses in REG when compared to RND patterns. This manifested from three tones after the onset of the pattern repetition, even in the context of slower sequences characterized by extended pattern durations (2,500 ms). This observation underscores the remarkable implicit sensitivity of the auditory brain to acoustic regularities. Importantly, brain responses evoked by single tones exhibited the opposite pattern-stronger responses to tones in RND than REG sequences. The demonstration of simultaneous but opposing sustained and evoked response effects reveals concurrent processes that shape the representation of unfolding auditory patterns.

Effects of continuous monoculture on rhizosphere soil nutrients, growth, physiological characteristics, hormone metabolome of Casuarina equisetifolia and their interaction analysis.

Continuous planting is unavoidable in agricultural production, but continuous planting affects plant growth and physiological characteristics. In this study, we analyzed rhizosphere soil nutrients, physiological characteristics, hormone metabolome changes and their interactions of Casuarina equisetifolia (C. equisetifolia) with the increase of continuous planting number. The results found that C. equisetifolia root was significantly inhibited, the plant height was dwarfed and the biomass was significantly reduced as continuous planting number increased. Secondly, continuous planting caused a decrease in the rhizosphere soil nutrient transformation capacity, and a significant decrease in the total soil nutrient and available nutrient content. Analysis of physiological indexes showed that continuous planting resulted in a decrease in nitrogen, phosphorus, and potassium content, a decrease in the activity of physiological indexes of resistance, and a decrease in photosynthetic capacity of C. equisetifolia leaves. Hormone metabolome analysis showed that continuous planting critically affected the accumulation of five characteristic hormones in C. equisetifolia leaves, in which salicylic acid 2-O-ß-glucoside (SAG), 2-oxindole-3-acetic acid (OxIAA), trans-zeatin-O-glucoside (tZOG) and gibberellin A3 (GA3) content decreased significantly while abscisic acid (ABA) content increased significantly. In conclusion, continuous planting lowered the rhizosphere soil nutrient transformation capacity of C. equisetifolia, lowered the soil available nutrient content, inhibited their root growth, and hindered the nutrient uptake and transportation by the root, thus led to the decrease of the nutrient accumulation capacity in the leaves of C. equisetifolia, and the decrease of SAG, OxIAA, and tZOG, GA3 synthesis ability decreased, ABA accumulated in large quantities, C. equisetifolia resistance and photosynthesis ability decreased, and their growth was impeded. This study provides insights for the effective management of continuous planting in the cultivation of C. equisetifolia.

Identification of genomic diversity and selection signatures in Luxi cattle using whole-genome sequencing data.

OBJECTIVE: The objective of this study was to investigate the genetic diversity, population structure and whole-genome selection signatures of Luxi cattle to reveal its genomic characteristics in terms of meat and carcass traits, skeletal muscle development, body size, and other traits. METHODS: To further analyze the genomic characteristics of Luxi cattle, this study sequenced the whole-genome of 16 individuals from the core conservation farm in Shandong region, and collected 174 published genomes of cattle for conjoint analysis. Furthermore, three different statistics (pi, Fst, and XP-EHH) were used to detect potential positive selection signatures related to selection in Luxi cattle. Moreover, gene ontology and Kyoto encyclopedia of genes and genomes pathway enrichment analyses were performed to reveal the potential biological function of candidate genes harbored in selected regions. RESULTS: The results showed that Luxi cattle had high genomic diversity and low inbreeding levels. Using three complementary methods (pi, Fst, and XP-EHH) to detect the signatures of selection in the Luxi cattle genome, there were 2,941, 2,221 and 1,304 potentially selected genes identified, respectively. Furthermore, there were 45 genes annotated in common overlapping genomic regions covered 0.723 Mb, including PLAG1 zinc finger (PLAG1), dedicator of cytokinesis 3 (DOCK3), ephrin A2 (EFNA2), DAZ associated protein 1 (DAZAP1), Ral GTPase activating protein catalytic subunit alpha 1 (RALGAPA1), mediator complex subunit 13 (MED13), and decaprenyl diphosphate synthase subunit 2 (PDSS2), most of which were enriched in pathways related to muscle growth and differentiation and immunity. CONCLUSION: In this study, we provided a series of genes associated with important economic traits were found in positive selection regions, and a scientific basis for the scientific conservation and genetic improvement of Luxi cattle.

Development and validation of prediction model for older adults with cognitive frailty.

OBJECTIVE: This study sought to develop and validate a 6-year risk prediction model in older adults with cognitive frailty (CF). METHODS: In the secondary analysis of Chinese Longitudinal Healthy Longevity Survey (CLHLS), participants from the 2011-2018 cohort were included to develop the prediction model. The CF was assessed by the Chinese version of Mini-Mental State Exam (CMMSE) and the modified Fried criteria. The stepwise regression was used to select predictors, and the logistic regression analysis was conducted to construct the model. The model was externally validated using the temporal validation method via the 2005-2011 cohort. The discrimination was measured by the area under the curve (AUC), and the calibration was measured by the calibration plot. A nomogram was conducted to vividly present the prediction model. RESULTS: The development dataset included 2420 participants aged 60 years or above, and 243 participants suffered from CF during a median follow-up period of 6.91 years (interquartile range 5.47-7.10 years). Six predictors, namely, age, sex, residence, body mass index (BMI), exercise, and physical disability, were finally used to develop the model. The model performed well with the AUC of 0.830 and 0.840 in the development and external validation datasets, respectively. CONCLUSION: The study could provide a practical tool to identify older adults with a high risk of CF early. Furthermore, targeting modifiable factors could prevent about half of the new-onset CF during a 6-year follow-up.

Associations of sarcopenia, sarcopenia parameters and motoric cognitive risk syndrome in Chinese older adults.

Background: Motoric cognitive risk syndrome (MCR) is a pre-dementia symptom strongly predicting cognitive decline and dementia. Although advancements in elucidating the epidemiology of MCR, the evidence about the association between sarcopenia, sarcopenia parameters, and MCR remains scarce. Objectives: The purpose of this study was to determine the associations between sarcopenia, sarcopenia parameters, and MCR among community-dwelling Chinese older adults. Methods: A total of 4,184 community-dwelling older adults from the China Health and Retirement Longitudinal Study (CHARLS) in the 2011 waves were included. Sarcopenia was diagnosed according to the Asian Working Group for Sarcopenia criteria. Sarcopenia parameters included handgrip strength (HGS), height-adjusted appendicular skeletal muscle mass (ASM/Ht2), and five-times sit-to-stand test time (FTSSTT). MCR was defined as subjective cognitive complaints and slow gait speed without dementia or impaired mobility. The associations between sarcopenia, sarcopenia parameters, and MCR were conducted using the logistic regression model. The restricted cubic spline with four knots were performed to determine the nonlinear and linear relationships between HGS, ASM/Ht2, FTSSTT, and MCR. Results: The prevalence of MCR in wave 2011 of CHARLS was 11.2%. After adjustment for potential confounders, we found sarcopenia [odd ratio (OR) (95% CI): 1.70 (1.13 ~ 2.54), p = 0.011], lower HGS [0.97 (0.96 ~ 0.99), p = 0.001], and more FTSSTT [1.12 (1.10 ~ 1.15), p < 0.001] were significantly associated with a higher risk of MCR. There was an inverse linear dose-response between HGS and MCR (p for overall = 0.008, p for nonlinearity =0.776). The nonlinear relationship between FTSSTT and MCR was found (p for overall <0.001, p for nonlinearity = 0.025) with FTSSTT ≥29 s being associated with a higher risk of MCR. A dose-response relationship was not found between ASM/Ht2 and MCR (p for overall =0.589). Conclusion: Sarcopenia, lower HGS, and higher FTSSTT are associated with MCR among older adults in China, while the latter two exhibit a dose-response relationship with MCR. It is suggested that timely identification and management of sarcopenia and its parameters may help delay the progression of cognitive impairment and promote healthy aging.

Metagenomics-based exploration of key soil microorganisms contributing to continuously planted Casuarina equisetifolia growth inhibition and their interactions with soil nutrient transformation.

Casuarina equisetifolia (C. equisetifolia) is an economically important forest tree species, often cultivated in continuous monoculture as a coastal protection forest. Continuous planting has gradually affected growth and severely restricted the sustainable development of the C. equisetifolia industry. In this study, we analyzed the effects of continuous planting on C. equisetifolia growth and explored the rhizosphere soil microecological mechanism from a metagenomic perspective. The results showed that continuous planting resulted in dwarfing, shorter root length, and reduced C. equisetifolia seedling root system. Metagenomics analysis showed that 10 key characteristic microorganisms, mainly Actinoallomurus, Actinomadura, and Mycobacterium, were responsible for continuously planted C. equisetifolia trees. Quantitative analysis showed that the number of microorganisms in these three genera decreased significantly with the increase of continuous planting. Gene function analysis showed that continuous planting led to the weakening of the environmental information processing-signal transduction ability of soil characteristic microorganisms, and the decrease of C. equisetifolia trees against stress. Reduced capacity for metabolism, genetic information processing-replication and repair resulted in reduced microbial propagation and reduced microbial quantity in the rhizosphere soil of C. equisetifolia trees. Secondly, amino acid metabolism, carbohydrate metabolism, glycan biosynthesis and metabolism, lipid metabolism, metabolism of cofactors and vitamins were all significantly reduced, resulting in a decrease in the ability of the soil to synthesize and metabolize carbon and nitrogen. These reduced capacities further led to reduced soil microbial quantity, microbial carbon and nitrogen, microbial respiration intensity, reduced soil enzyme nutrient cycling and resistance-related enzyme activities, a significant reduction in available nutrient content of rhizosphere soils, a reduction in the ion exchange capacity, and an impediment to C. equisetifolia growth. This study provides an important basis for the management of continuously planted C. equisetifolia plantations.

Bidirectional associations between hearing difficulty and cognitive function in Chinese adults: a longitudinal study.

Background: Middle-aged and older adults frequently experience hearing loss and a decline in cognitive function. Although an association between hearing difficulty and cognitive function has been demonstrated, its temporal sequence remains unclear. Therefore, we investigated whether there are bidirectional relationships between hearing difficulty and cognitive function and explored the mediating role of depressive symptoms in this relationship. Method: We used the cross-lagged panel model and the random-intercept cross-lagged panel model to look for any possible two-way link between self-reported hearing difficulty and cognitive function. To investigate depressive symptoms' role in this association, a mediation analysis was conducted. The sample was made up of 4,363 adults aged 45 and above from the China Health and Retirement Longitudinal Study (CHARLS; 2011-2018; 44.83% were women; mean age was 56.16 years). One question was used to determine whether someone had a hearing impairment. The tests of cognitive function included episodic memory and intelligence. The Center for Epidemiologic Studies Depression Scale, which consists of 10 items, was used to measure depressive symptoms. Results: A bidirectional association between hearing and cognition was observed, with cognition predominating (Wald χ2 (1) = 7.241, p < 0.01). At the between-person level, after controlling for potential confounders, worse hearing in 2011 predicted worse cognitive function in 2013 (ß = -0.039, p < 0.01) and vice versa (ß = -0.041, p < 0.01) at the between-person level. Additionally, there was no corresponding cross-lagged effect of cognitive function on hearing difficulty; rather, the more hearing difficulty, the greater the cognitive decline at the within-person level. According to the cross-lagged mediation model, depressive symptoms partially mediates the impact of cognitive function on subsequent hearing difficulty (indirect effect: -0.003, bootstrap 95% confidence interval: -0.005, -0.001, p < 0.05), but not the other way around. Conclusion: These results showed that within-person relationships between hearing impairment and cognitive function were unidirectional, while between-person relationships were reciprocal. Setting mental health first may be able to break the vicious cycle that relates hearing loss to cognitive decline. Comprehensive long-term care requires services that address depressive symptoms and cognitive decline to be integrated with the hearing management.

Effect of continuous planting on Casuarina equisetifolia rhizosphere soil physicochemical indexes, microbial functional diversity and metabolites.

Continuous planting has a severe impact on the growth of Casuarina equisetifolia. In this study, the effects of three different long-term monocultures (one, two and three replanting) on the physicochemical indexes, microbial functional diversity, and soil metabolomics were analyzed in C. equisetifolia rhizosphere soil. The results showed that rhizosphere soil organic matter content, cation exchange capacity, total and available nitrogen, total and available phosphorus, and total and available potassium contents significantly decreased with the increasing number of continuous plantings. The evaluation of microbial functional diversity revealed a reduction in the number of soil microorganisms that rely on carbohydrates for carbon sources and an increase in soil microorganisms that used phenolic acid, carboxylic acid, fatty acid, and amines as carbon sources. Soil metabolomics analysis showed a significant decrease in soil carbohydrate content and a significant accumulation of autotoxic acid, amine, and lipid in the C. equisetifolia rhizosphere soil. Consequently, the growth of C. equisetifolia could hinder total nutrient content and their availability. Thus, valuable insights for managing the cultivation of C. equisetifolia and soil remediation were provided.

Impact of hearing loss on cognitive function in community-dwelling older adults: serial mediation of self-rated health and depressive anxiety symptoms.

Background: Hearing loss can exacerbate cognitive decline; therefore, exploring the mechanisms through which hearing loss affects cognitive function is crucial. The current study aimed to investigate the impact of hearing loss on cognitive function and the mediating role played by self-rated health and depressive anxiety symptoms. Methods: Using stratified whole-group random sampling, the study employed a cross-sectional design and included 624 participants aged ≥65 years from three communities in Urumqi, China. Cognitive function was assessed using the Mini-Mental State Examination. Hearing function and self-rated health were determined by self-report. The 15-item Geriatric Depression Scale and the 7-item Generalized Anxiety Disorder Scale were used to assess depressive anxiety symptoms. Serial mediation analysis was performed using AMOS 26.0. Results: Hearing loss can not only negatively affect cognitive function in older adults directly (direct effect = -0.106; SE = 0.045; 95% confidence interval (CI): -0.201 to -0.016), but also indirectly affect the relationship between hearing loss and cognitive function through self-rated health and depressive anxiety symptoms. The results of the serial mediation analysis showed that the total indirect effect of self-rated health and depressive anxiety symptoms was -0.115 (95% CI: -0.168 to -0.070), and the total effect of the model was -0.221 (95% CI: -0.307 to -0.132), with the total indirect effect accounting for 52.04% of the total effect of the model. Conclusion: Our study discovered that there is a partial mediation of the relationship between hearing loss and cognitive function by self-rated health and depressive anxiety symptoms. It is suggested that by enhancing self-rated health and ensuring good mental health, the decline in cognitive function among older adults with hearing loss can be delayed.

Detection of genomic variations and selection signatures in Wagyu using whole-genome sequencing data.

Wagyu is recognized for producing marbled beef with high nutritional value and flavor. Reportedly, Wagyu has been widely used to improve the meat quality of local breeds around the world. However, studies on the genetic mechanism of meat quality in Wagyu at the whole-genome level are rarely reported. Here, whole-genome sequencing data of 11 Wagyu and 115 other individuals were used to explore the genomic variations and genes under selection pressure in Wagyu. A total of 31 349 non-synonymous variants and 53 102 synonymous variants were identified in Wagyu. The population structure analysis showed that Wagyu had the closest genetic relationship with Mishima-Ushi cattle and was apparently separated from other cattle breeds. Then, composite likelihood ratio (CLR), integrated haplotype score, fixation index and cross-population composite likelihood ratio (XP-CLR) tests were performed to identify the candidate genes under positive selection in Wagyu. In total, 770 regions containing 312 genes were identified by at least three methods. Among them, 97 regions containing 27 genes were detected by all four methods. We specifically illustrate a list of interesting genes, including LRP2BP, GAA, CACNG6, CXADR, GPCPD1, KLF2, KLF13, SOX5, MYBPC1, SLC25A10, ATP8A1 and MYH15, which are associated with lipid metabolism, fat deposition, muscle development, bone development, feed intake and growth traits in Wagyu. This is the first study to explore the genomic variations and selection signatures of Wagyu at the whole-genome level. These results will provide significant help to beef cattle improvement and breeding.

Effects of choline deficiency and supplementation on lipid droplet accumulation in bovine primary liver cells in vitro.

Rumen-protected choline (RPC) supplementation in the periparturient period has in some instances prevented and alleviated fatty liver disease in dairy cows. Mechanistically, however, it is unclear how choline prevents the accumulation of lipid droplets (LD) in liver cells. In this study, primary liver cells isolated from liver tissue obtained via puncture biopsy from 3 nonpregnant mid-lactation multiparous Holstein cows (∼160 d postpartum) were used. Analyses of LD via oil red O staining, protein abundance via Western blotting, and phospholipid content and composition measured by thin-layer chromatography and HPLC/mass spectrometry were performed in liver cells cultured in choline-deficient medium containing 150 µmol/L linoleic acid for 24 h. In a subsequent experiment, lipophagy was assessed in liver cells cultured with 30, 60, or 90 µmol/L choline-chloride. All data were analyzed statistically using SPSS 20.0 via t-tests or one-way ANOVA. Compared with liver cells cultured in Dulbecco's Modified Eagle Medium alone, choline deficiency increased the average diameter of LD (1.59 vs. 2.10 µm), decreased the proportion of small LD (<2 µm) from 75.3% to 56.6%, and increased the proportion of large LD (>4 µm) from 5.6% to 15.0%. In addition, the speed of LD fusion was enhanced by the absence of choline. Among phospholipid species, the phosphatidylcholine (PC) content of liver cells decreased by 34.5%. Seventeen species of PC (PC [18:2_22:6], PC [15:0_16:1], PC [14:0_20:4], and so on) and 6 species of lysophosphatidylcholine (LPC; LPC [15:0/0:0]), PC (22:2/0:0), LPC (20:2/0:0), and so on] were decreased, while PC (14:1_16:1) and LPC (0:0/20:1) were increased. Choline deficiency increased the triglyceride (TAG) content (0.57 vs. 0.39 µmol/mg) in liver cells and increased the protein abundance of sterol regulatory element binding protein 1, sterol regulatory element binding protein cleavage activation protein, and fatty acid synthase by 23.5%, 17%, and 36.1%, respectively. Upon re-supplementation with choline, the phenotype of LD (TAG content, size, proportion, and phospholipid profile) was reversed, and the ratio of autophagy marker LC3II/LC3I protein was significantly upregulated in a dose-dependent manner. Overall, at least in vitro in mid-lactation cows, these data demonstrated that PC synthesis is necessary for normal LD formation, and both rely on choline availability. According to the limitation of the source of liver cells used, further work should be conducted to ascertain that these effects are applicable to liver cells from postpartum cows, the physiological stage where the use of RPC has been implemented for the prevention and treatment of fatty liver.

Proanthocyanidin A2 attenuates the activation of hepatic stellate cells by activating the PPAR-γ signalling pathway.

Liver fibrosis is the pathological process of chronic liver diseases induced by hepatic stellate cells. Proanthocyanidin A2 (PA2) has multiple pharmacological activities. In this study, we aimed to explore the effects of PA2 on hepatic stellate cell (HSC) activation in liver fibrosis. LX-2 cells were treated with TGF-ß1 to establish a fibrosis cell model. Cell viability was evaluated using cell counting kit-8. The levels of fibrosis-related factors (collagen I, fibronectin, and α-SMA) were examined using quantitative real-time polymerase chain reaction, western blot, and immunofluorescence assay. The molecular mechanisms of PA2 were evaluated by RNA-seq, bioinformatic analysis, and western blot. The results showed that PA2 suppressed cell viability, and downregulated fibrosis-related factors induced by TGF-ß1, suggesting PA2 suppressed the activation of HSCs. PA2 treatment-induced differentially expressed mRNAs are predicted to be associated with the PPAR-γ pathway. PA2 reversed the downregulation of PPAR-γ and the upregulation of phosphorylated (p)-Smad2 and Smad3. A rescue experiment illustrated that the inactivation of the PPAR-γ pathway reversed the effects of PA2 on cell viability and HSC activation. In conclusion, PA2 inhibited TGF-ß1-induced activation of HSCs by activating the PPAR-γ/Smad pathway. The findings suggested that PA2 may be an effective treatment for liver fibrosis.

Delayed gadolinium contrast-enhanced 3D-T1 SPACE STIR sequence can better visualize abnormal cranial nerves: a case report.

With the advancement of magnetic resonance imaging (MRI) techniques, general radiographic methods are no longer sufficient for accurately displaying the structure and pathway of cranial nerves. Various sequences, including 3-dimensional sampling perfection with application-optimized contrast using different flip angle evolution (SPACE), have been developed through MRI technology to effectively display the location and severity of damaged cranial nerves. This current case report describes a 36-year-old male patient with multiple cranial nerve injuries resulting from an invasive Mucor infection. While performing MRI scanning on this patient, a 1-h delayed enhanced MRI 3D-T1 SPACE short tau inversion recovery (STIR) sequence proved more effective in eliminating background interference and assessing neurological damage with greater clarity than conventional enhancement methods. This approach may prove beneficial in accurately evaluating the extent of cranial neuropathy, thus facilitating clinical applications.

Identification of an Arylnaphthalene Lignan Derivative as an Inhibitor against Dengue Virus Serotypes 1 to 4 (DENV-1 to -4) Using a Newly Developed DENV-3 Infectious Clone and Replicon.

Dengue virus (DENV) is the most widespread arbovirus, causing symptoms ranging from dengue fever to severe dengue, including hemorrhagic fever and shock syndrome. Four serotypes of DENV (DENV-1 to -4) can infect humans; however, no anti-DENV drug is available. To facilitate the study of antivirals and viral pathogenesis, here we developed an infectious clone and a subgenomic replicon of DENV-3 strains for anti-DENV drug discovery by screening a synthetic compound library. The viral cDNA was amplified from a serum sample from a DENV-3-infected individual during the 2019 epidemic; however, fragments containing the prM-E-partial NS1 region could not be cloned until a DENV-3 consensus sequence with 19 synonymous substitutions was introduced to reduce putative Escherichia coli promoter activity. Transfection of the resulting cDNA clone, plasmid DV3syn, released an infectious virus titer of 2.2 × 102 focus-forming units (FFU)/mL. Through serial passages, four adaptive mutations (4M) were identified, and addition of 4M generated recombinant DV3syn_4M, which produced viral titers ranging from 1.5 × 104 to 6.7 × 104 FFU/mL and remained genetically stable in transformant bacteria. Additionally, we constructed a DENV-3 subgenomic replicon and screened an arylnaphthalene lignan library, from which C169-P1 was identified as exhibiting inhibitory effects on viral replicon. A time-of-drug addition assay revealed that C169-P1 also impeded the internalization process of cell entry. Furthermore, we demonstrated that C169-P1 inhibited the infectivity of DV3syn_4M, as well as DENV-1, DENV-2, and DENV-4, in a dose-dependent manner. This study provides an infectious clone and a replicon for the study of DENV-3 and a candidate compound for future development against DENV-1 to -4 infections. IMPORTANCE Dengue virus (DENV) is the most prevalent mosquito-transmitted virus, and there is no an anti-dengue drug. Reverse genetic systems representative of different serotype viruses are invaluable tools for the study of viral pathogenesis and antiviral drugs. Here, we developed an efficient infectious clone of a clinical DENV-3 genotype III isolate. We successfully overcame the instability of flavivirus genome-length cDNA in transformant bacteria, an unsolved issue for construction of cDNA clones of flaviviruses, and adapted this clone to efficiently produce infectious viruses following plasmid transfection of cell culture. Moreover, we constructed a DENV-3 subgenomic replicon and screened a compound library. An arylnaphthalene lignan, C169-P1, was identified as an inhibitor of virus replication and cell entry. Finally, we demonstrated that C169-P1 exhibited a broad-spectrum antiviral effect against the infections with DENV-1 to -4. The reverse genetic systems and the compound candidate described here facilitate the study of DENV and related RNA viruses.