Multi-biobank Mendelian randomization analyses identify opposing pathways in plasma low-density lipoprotein-cholesterol lowering and gallstone disease.

Plasma low-density lipoprotein (LDL)-cholesterol is positively associated with coronary artery disease risk while biliary cholesterol promotes gallstone formation. Different plasma LDL-cholesterol lowering pathways may have distinct effects on biliary cholesterol and thereby gallstone disease risk. We conducted a Mendelian randomization (MR) study using data from the UK Biobank (30,547 gallstone disease cases/336,742 controls), FinnGen (34,461 cases/301,383 controls) and Biobank Japan (9,305 cases/168,253 controls). We first performed drug-target MR analyses substantiated by colocalization to investigate the effects of plasma LDL-cholesterol lowering therapies on gallstone disease risk. We then performed clustered MR analyses and pathway analyses to identify distinct mechanisms underlying the association of plasma LDL-cholesterol with gallstone disease risk. For a 1-standard deviation reduction in plasma LDL-cholesterol, genetic mimics of statins were associated with lower gallstone disease risk (odds ratio 0.72 [95% confidence interval 0.62, 0.83]), but genetic mimics of PCSK9 inhibitors and targeting apolipoprotein B were associated with higher risk (1.11 [1.03, 1.19] and 1.23 [1.13, 1.35]). The association for statins was supported by colocalization (posterior probability 98.7%). Clustered MR analyses identified variant clusters showing opposing associations of plasma LDL-cholesterol with gallstone disease risk, with some evidence for ancestry-and sex-specific associations. Among variants lowering plasma LDL-cholesterol, those associated with lower gallstone disease risk were mapped to glycosphingolipid biosynthesis pathway, while those associated with higher risk were mapped to pathways relating to plasma lipoprotein assembly, remodelling, and clearance and ATP-binding cassette transporters. This MR study provides genetic evidence that different plasma LDL-cholesterol lowering pathways have opposing effects on gallstone disease risk.

Dose-Response Associations of Lipid Traits With Coronary Artery Disease and Mortality.

Importance: Apolipoprotein B (apoB), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG) are associated with coronary artery disease (CAD). However, trial evidence for the association of intensive LDL-C lowering and TG lowering with mortality is less definitive. Objectives: To investigate the associations of apoB, LDL-C, and TG with CAD and mortality, both overall and by sex and age, and to characterize the shapes of these associations. Design, Setting, and Participants: This genetic association study used linear and nonlinear mendelian randomization (MR) to analyze a population-based cohort of individuals of European ancestry from the UK Biobank, which recruited participants from 2006 to 2010 with follow-up information updated until September 2021. Data analysis occurred from December 2022 to November 2023. Exposures: Genetically predicted apoB, LDL-C, and TG. Main Outcomes and Measures: The primary outcomes were CAD, all-cause mortality, and cause-specific mortality. Genetic associations with CAD were calculated using logistic regression, associations with all-cause mortality using Cox proportional hazards regression, and associations with cause-specific mortality using cause-specific Cox proportional hazards regression with censoring for other causes of mortality. Results: This study included 347 797 participants (mean [SD] age, 57.2 [8.0] years; 188 330 female [54.1%]). There were 23 818 people who developed CAD and 23 848 people who died. Genetically predicted apoB was positively associated with risk of CAD (odds ratio [OR], 1.65 per SD increase; 95% CI 1.57-1.73), all-cause mortality (hazard ratio [HR], 1.11; 95% CI, 1.06-1.16), and cardiovascular mortality (HR, 1.36; 95% CI, 1.24-1.50), with some evidence for larger associations in male participants than female participants. Findings were similar for LDL-C. Genetically predicted TG was positively associated with CAD (OR, 1.60; 95% CI 1.52-1.69), all-cause mortality (HR, 1.08; 95% CI, 1.03-1.13), and cardiovascular mortality (HR, 1.21; 95% CI, 1.09-1.34); however, sensitivity analyses suggested evidence of pleiotropy. The association of genetically predicted TG with CAD persisted but it was no longer associated with mortality outcomes after controlling for apoB. Nonlinear MR suggested that all these associations were monotonically increasing across the whole observed distribution of each lipid trait, with no diminution at low lipid levels. Such patterns were observed irrespective of sex or age. Conclusions and relevance: In this genetic association study, apoB (or, equivalently, LDL-C) was associated with increased CAD risk, all-cause mortality, and cardiovascular mortality, all in a dose-dependent way. TG may increase CAD risk independent of apoB, although the possible presence of pleiotropy is a limitation. These insights highlight the importance of apoB (or, equivalently, LDL-C) lowering for reducing cardiovascular morbidity and mortality across its whole distribution.

Reduced attention on restricted organochlorine pesticides, whereas still noteworthy of the impact on the deep soil and groundwater: a historical site study in southern China.

The use of hexachlorocyclohexanes (HCHs) in pesticides has been prohibited for decades in China. Since then, there have been urbanization and transformation of the functional areas of many sites, which were formerly involved in the HCH industry. However, it is possible that, unless properly managed, these sites may still contain HCH residues in the soil and thus pose a threat to the surrounding environment and the quality of groundwater. This study aimed to characterize soil residues in a typical site that was historically involved in HCH production in southern China, by analyzing the α-HCH, ß-HCH, and γ-HCH contents of the soil. The results suggested that HCHs persist in the environment and can have long-term effects. It was found that α-HCH and ß-HCH were present in many samples in concentrations that were comparable or higher than those specified by China's Class 1 screening values. The distribution of residues was significantly correlated with the historical HCH production activities in the areas. The characteristic ratios of α-HCH/γ-HCH and ß-HCH/(α + γ)-HCH at different soil depths were 1.4-3.7 and 0.21-1.04, respectively, which indicated the presence of significant localized residues of HCHs. The presence of HCHs in the soil suggested a downward migration, with concentrations rapidly decreasing in the upper layer soil (0-5 m), but a gradual increase in the deeper soil (5-14 m). HCHs were detected at depths exceeding 24 m, indicating heavy penetration. The proportions of γ-HCH and ß-HCH changed with increasing soil depth, which was related to their relatively volatile and stable molecular structures, respectively. The results strongly suggested that there is widespread contamination of both soil and groundwater by HCHs even after decades. The likelihood of residual HCHs in the soil should therefore be taken into full consideration during urban planning to limit risks to human and environmental health.

Genetically mimicked effects of ASGR1 inhibitors on all-cause mortality and health outcomes: a drug-target Mendelian randomization study and a phenome-wide association study.

BACKGROUND: Asialoglycoprotein receptor 1 (ASGR1) is emerging as a potential drug target to reduce low-density lipoprotein (LDL)-cholesterol and coronary artery disease (CAD) risk. Here, we investigated genetically mimicked ASGR1 inhibitors on all-cause mortality and any possible adverse effects. METHODS: We conducted a drug-target Mendelian randomization study to assess genetically mimicked effects of ASGR1 inhibitors on all-cause mortality and 25 a priori outcomes relevant to lipid traits, CAD, and possible adverse effects, i.e. liver function, cholelithiasis, adiposity and type 2 diabetes. We also performed a phenome-wide association study of 1951 health-related phenotypes to identify any novel effects. Associations found were compared with those for currently used lipid modifiers, assessed using colocalization, and replicated where possible. RESULTS: Genetically mimicked ASGR1 inhibitors were associated with a longer lifespan (3.31 years per standard deviation reduction in LDL-cholesterol, 95% confidence interval 1.01 to 5.62). Genetically mimicked ASGR1 inhibitors were inversely associated with apolipoprotein B (apoB), triglycerides (TG) and CAD risk. Genetically mimicked ASGR1 inhibitors were positively associated with alkaline phosphatase, gamma glutamyltransferase, erythrocyte traits, insulin-like growth factor 1 (IGF-1) and C-reactive protein (CRP), but were inversely associated with albumin and calcium. Genetically mimicked ASGR1 inhibitors were not associated with cholelithiasis, adiposity or type 2 diabetes. Associations with apoB and TG were stronger for ASGR1 inhibitors compared with currently used lipid modifiers, and most non-lipid effects were specific to ASGR1 inhibitors. The probabilities for colocalization were > 0.80 for most of these associations, but were 0.42 for lifespan and 0.30 for CAD. These associations were replicated using alternative genetic instruments and other publicly available genetic summary statistics. CONCLUSIONS: Genetically mimicked ASGR1 inhibitors reduced all-cause mortality. Beyond lipid-lowering, genetically mimicked ASGR1 inhibitors increased liver enzymes, erythrocyte traits, IGF-1 and CRP, but decreased albumin and calcium.

Investigating sex-specific associations of lipid traits with type 2 diabetes, glycemic traits and sex hormones using Mendelian randomization.

BACKGROUND: Low-density lipoprotein (LDL)-cholesterol is positively associated with cardiovascular disease (CVD) and inversely associated with type 2 diabetes, which could detract from lipid modification. Here, we examined whether lipid traits potentially relevant to CVD aetiology, i.e. apolipoprotein B (apoB), triglycerides (TG) and lipoprotein(a) [Lp(a)] exhibited the same associations. We investigated sex-specifically, including the role of sex hormones, because sex disparities exist in lipid profile and type 2 diabetes. We also replicated where possible. METHODS: We used Mendelian randomization (MR) to examine sex-specific associations of apoB, TG and Lp(a) with type 2 diabetes, HbA1c, fasting insulin, fasting glucose, testosterone and estradiol in the largest relevant sex-specific genome-wide association studies (GWAS) in people of European ancestry and replicated where possible. We also assessed sex-specific associations of liability to type 2 diabetes with apoB, TG and Lp(a). RESULTS: Genetically predicted apoB and Lp(a) had little association with type 2 diabetes or glycemic traits in women or men. Genetically predicted higher TG was associated with higher type 2 diabetes risk [odds ratio (OR) 1.44 per standard deviation (SD), 95% confidence interval (CI) 1.26 to 1.65], HbA1c and fasting insulin specifically in women. Higher TG was associated with lower testosterone in women and higher testosterone in men, but with lower estradiol in men and women. Genetic liability to type 2 diabetes was associated with higher TG in women, and possibly with lower apoB in men. CONCLUSIONS: Lipid traits potentially relevant to CVD aetiology do not exhibit contrasting associations with CVD and type 2 diabetes. However, higher TG is associated with higher type 2 diabetes risk and glycemic traits, which in turn further increases TG specifically in women, possibly driven by sex hormones.

Statins, Type 2 Diabetes, and Body Mass Index: A Univariable and Multivariable Mendelian Randomization Study.

CONTEXT: Statins and possibly other lipid modifiers increase type 2 diabetes risk and body mass index (BMI). However, to what extent BMI mediates the diabetogenic effects of lipid modifiers remains unclear. OBJECTIVE: We used Mendelian randomization (MR) to investigate the effects of commonly used lipid modifiers on type 2 diabetes risk and glycemic traits, and any mediation by BMI. METHODS: Using established genetic variants to mimic commonly used lipid modifiers (ie, statins, PCSK9 inhibitors, and ezetimibe), we assessed their associations with type 2 diabetes risk, glycated hemoglobin (HbA1c), fasting insulin, fasting glucose, and BMI in the largest relevant genome-wide association studies (GWAS) in people of European ancestry, and where possible, in East Asians. We used multivariable MR to examine the role of lipid modifiers independent of BMI. RESULTS: Genetically mimicked effects of statins and ezetimibe, but not PCSK9 inhibitors were associated with higher risk of type 2 diabetes (odds ratio [OR] 1.74 [95% CI, 1.49 to 2.03]; 1.92 [1.22 to 3.02]; 1.06 [0.87 to 1.29] per SD reduction in low-density lipoprotein (LDL)-cholesterol). Of these lipid modifiers, only genetic mimics of statins were associated with higher BMI (0.33 SD [0.29 to 0.38] per SD reduction in LDL-cholesterol), which explained 54% of the total effect of statins on type 2 diabetes risk. CONCLUSION: Higher BMI mediated more than half of the diabetogenic effects of statins, which did not extend to other commonly used lipid modifiers. Further investigations are needed to clarify drug-specific mechanisms underlying the effects of lipid modifiers on type 2 diabetes.

Sex differences in the association of fasting glucose with HbA1c, and their consequences for mortality: A Mendelian randomization study.

BACKGROUND: Hemoglobin A1c (HbA1c) is used for diabetes diagnosis and management. HbA1c also represents iron-related erythrocyte properties which differ by sex. We investigated erythrocyte properties on HbA1c and glucose, and whether corresponding consequences for mortality differed by sex. METHODS: In this two-sample Mendelian randomization study using the largest publicly available European descent summary statistics, we assessed sex-specific associations of iron (n=163,511) and hemoglobin (188,076 women/162,398 men) with HbA1c (185,022 women/159,160 men) and fasting glucose (73,089 women/67,506 men), of fasting glucose with HbA1c and diabetes (cases=6,589 women/10,686 men, controls=187,137 women/155,780 men), and of fasting glucose (n=140,595), HbA1c (n=146,806) and liability to diabetes (74,124 cases/824,006 controls) with parental attained age (412,937 mothers/415,311 fathers). FINDINGS: Iron and hemoglobin were inversely associated with HbA1c but not fasting glucose. Fasting glucose was more strongly associated with HbA1c and diabetes in women (1.65 standard deviation (SD) per mmol/L [95% confidence interval 1.58, 1.72]; odds ratio (OR) 7.36 per mmol/L [4.12, 10.98]) than men (0.89 [0.81, 0.98]; OR 2.79 [1.96, 4.98]). The inverse associations of HbA1c and liability to diabetes with lifespan were possibly stronger in men (-1.80 years per percentage [-2.77, -0.42]; -0.93 years per logOR [-1.23, -0.59]) than women (-0.80 [-2.69, 0.66]; -0.44 [-0.62, -0.26]). INTERPRETATION: HbA1c underestimates fasting glucose in men compared with women, possibly due to erythrocyte properties. Whether HbA1c and liability to diabetes reduce lifespan more in men than women because diagnostic and management criteria involving HbA1c mean that glycemia in men is under-treated compared to women needs urgent investigation. FUNDING: None.

Investigating genetically mimicked effects of statins via HMGCR inhibition on immune-related diseases in men and women using Mendelian randomization.

Statins have been suggested as a potential treatment for immune-related diseases. Conversely, statins might trigger auto-immune conditions. To clarify the role of statins in allergic diseases and auto-immune diseases, we conducted a Mendelian randomization (MR) study. Using established genetic instruments to mimic statins via 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) inhibition, we assessed the effects of statins on asthma, eczema, allergic rhinitis, rheumatoid arthritis (RA), psoriasis, type 1 diabetes, systemic lupus erythematosus (SLE), multiple sclerosis (MS), Crohn's disease and ulcerative colitis in the largest available genome wide association studies (GWAS). Genetically mimicked effects of statins via HMGCR inhibition were not associated with any immune-related diseases in either study after correcting for multiple testing; however, they were positively associated with the risk of asthma in East Asians (odds ratio (OR) 2.05 per standard deviation (SD) decrease in low-density lipoprotein cholesterol (LDL-C), 95% confidence interval (CI) 1.20 to 3.52, p value 0.009). These associations did not differ by sex and were robust to sensitivity analysis. These findings suggested that genetically mimicked effects of statins via HMGCR inhibition have little effect on allergic diseases or auto-immune diseases. However, we cannot exclude the possibility that genetically mimicked effects of statins via HMGCR inhibition might increase the risk of asthma in East Asians.

Career choice regret during COVID-19 among healthcare students and professionals in mainland China: a cross-sectional study.

BACKGROUND: The COVID-19 epidemic affected the career choice of healthcare professionals and students. Career choice regret of healthcare professionals and students during COVID-19 outbreak and its affected factors are largely unexplored. METHODS: Convenience sample of nurses, doctors, and medical students were recruited from hospitals and universities nationwide. The data collected including demographic information, professional value before and after the COVID-19 outbreak, the Connor-Davidson Resilience Scale, and career choice regret level by an online questionnaire. Multinominal logistic regression was employed to explore the factors associated with career choice regret. RESULTS: In total, 9322 participants of convenience sampling were enrolled in, including 5786 nurses, 1664 doctors, and 1872 medical students. 6.7% participants had career choice regret. Multinominal logistic regression analysis showed, compared to participants with no regret, that as levels of psychological resilience increased, the odds of experiencing career choice regret decreased (OR = 0.95, 95% CI 0.94-0.96), while participants with lower professional value evaluation after the COVID-19 outbreak had higher probability to experience career choice regret (OR = 1.55,95% CI 1.50-1.61). Medical students were more likely to regret than nurses (OR = 1.65,95% CI 1.20-2.28), participants whose career/major choice was not their personal ideal had higher risk of experience career choice regret (OR = 1.59,95% CI 1.29-1.96), while participants who were very afraid of the coronavirus had higher risk to experience career choice regret then participants with no fear at all (OR = 2.00,95% CI 1.24-3.21). As for the medical students, results indicated that medical students major in nursing and undergraduates had higher risk to experience career choice regret compared to medical students major in clinical medicine and postgraduate (Master or PhD), with an odds ratios of 2.65(95% CI 1.56-4.49) and 6.85 (95% CI 2.48-18.91)respectively. CONCLUSIONS: A minority of healthcare professionals and medical students regretted their career choices during the COVID-19 outbreak. Enhance personal psychological resilience and professional value would helpful to reduce career choice regret among healthcare professionals and students during pandemic.

Glucagon Potentiates Insulin Secretion Via ß-Cell GCGR at Physiological Concentrations of Glucose.

Incretin-potentiated glucose-stimulated insulin secretion (GSIS) is critical to maintaining euglycemia, of which GLP-1 receptor (GLP-1R) on ß-cells plays an indispensable role. Recently, α-cell-derived glucagon but not intestine-derived GLP-1 has been proposed as the critical hormone that potentiates GSIS via GLP-1R. However, the function of glucagon receptors (GCGR) on ß-cells remains elusive. Here, using GCGR or GLP-1R antagonists, in combination with glucagon, to treat single ß-cells, α-ß cell clusters and isolated islets, we found that glucagon potentiates insulin secretion via ß-cell GCGR at physiological but not high concentrations of glucose. Furthermore, we transfected primary mouse ß-cells with RAB-ICUE (a genetically encoded cAMP fluorescence indicator) to monitor cAMP level after glucose stimulation and GCGR activation. Using specific inhibitors of different adenylyl cyclase (AC) family members, we revealed that high glucose concentration or GCGR activation independently evoked cAMP elevation via AC5 in ß-cells, thus high glucose stimulation bypassed GCGR in promoting insulin secretion. Additionally, we generated ß-cell-specific GCGR knockout mice which glucose intolerance was more severe when fed a high-fat diet (HFD). We further found that ß-cell GCGR activation promoted GSIS more than GLP-1R in HFD, indicating the critical role of GCGR in maintaining glucose homeostasis during nutrient overload.

Micro-ocean-bottom electromagnetic receiver for controlled-source electromagnetic and magnetotelluric data acquisition.

A new micro-ocean-bottom electromagnetic (EM) receiver is presented for marine magnetotelluric (MT) and controlled-source EM (CSEM) data acquisition, avoiding the bulkiness, high cost, high power consumption, and narrow bandwidth of the existing ocean-bottom EM receivers. The efficient, compact, and easy-to-use data logger employs a single 17-in. spherical glass as a float and uses an acoustic telemetry modem unit to control the burn-wire release of the seafloor instrument via a smartphone. The receiver design involves two induction coils for the magnetometer and four 8-m-span electric-field sensors for MT and CSEM data acquisition. The new ocean-bottom EM receiver was tested in sea. Compared with other miniaturized seabed EM receivers, the proposed receiver achieves better comprehensive performance for MT and CSEM data acquisition and meets the requirements of deep-mantle-structure research and seabed resource exploration.

Hyperglycemia-Induced Dysregulated Fusion Intermediates in Insulin-Secreting Cells Visualized by Super-Resolution Microscopy.

Impaired insulin release is a hallmark of type 2 diabetes and is closely related to chronically elevated glucose concentrations, known as "glucotoxicity." However, the molecular mechanisms by which glucotoxicity impairs insulin secretion remain poorly understood. In addition to known kiss-and-run and kiss-and-stay fusion events in INS-1 cells, ultrafast Hessian structured illumination microscopy (Hessian SIM) enables full fusion to be categorized according to the newly identified structures, such as ring fusion (those with enlarged pores) or dot fusion (those without apparent pores). In addition, we identified four fusion intermediates during insulin exocytosis: initial pore opening, vesicle collapse, enlarged pore formation, and final pore dilation. Long-term incubation in supraphysiological doses of glucose reduced exocytosis in general and increased the occurrence of kiss-and-run events at the expense of reduced full fusion. In addition, hyperglycemia delayed pore opening, vesicle collapse, and enlarged pore formation in full fusion events. It also reduced the size of apparently enlarged pores, all of which contributed to the compromised insulin secretion. These phenotypes were mostly due to the hyperglycemia-induced reduction in syntaxin-1A (Stx-1A) and SNAP-25 protein, since they could be recapitulated by the knockdown of endogenous Stx-1A and SNAP-25. These findings suggest essential roles for the vesicle fusion type and intermediates in regulating insulin secretion from pancreatic beta cells in normal and disease conditions.

Eating Habits and Lifestyles during the Initial Stage of the COVID-19 Lockdown in China: A Cross-Sectional Study.

Due to the outbreak of coronavirus disease 2019 (COVID-19), the Chinese government implemented strict lockdown measures to control the spread of infection. The impact of the COVID-19 lockdown on eating habits and lifestyles in the general population is unclear. This cross-sectional study was conducted via an online survey to obtain an overview of the food access, food intake, and physical activity of Chinese residents during the initial stage of the COVID-19 lockdown, and to investigate the association between staying at home/working from home and changes in eating habits and lifestyles. A total of 2702 participants (70.7% women) were included. Most of the participants maintained their habitual diet, while 38.2% increased their snack intake, 54.3% reported reduced physical activity, and 45.5% had increased sleep duration. Most people (70.1%) reported no change in body weight, while 25.0% reported an increase. Always staying at home/working from home was associated with an increase in animal product, vegetable, fruit, mushroom, nut, water, and snack intake, as well as sleep duration and frequency of skipping breakfast (odds ratio (OR) 1.54, 1.62, 1.58, 1.53, 1.57, 1.52, 1.77, 2.29, and 1.76 respectively). Suggestions should be made to encourage people to reduce their snack intake, maintain the daily consumption of breakfast, and increase physical activity during future lockdown periods.

Genitalia-related nursing embarrassment and its associated factors among female nurses in mainland China: a nationwide cross-sectional study.

BACKGROUND: Genital-related care is a common nursing procedure and may cause embarrassment for nurses. However, nurses' level of embarrassment and the factors associated with embarrassment have been scarcely studied. Therefore, a cross-sectional study was conducted to investigate genitalia-related care's embarrassment and its associated factors among Chinese female nurses. METHODS: Online questionnaires regarding the frequency of genitalia-related care and the embarrassment level were distributed to female nurses from the gynecology and urology departments between October and December 2019. Participants also completed the Chinese version of the professional identity scale for nurses and the Jefferson scale of empathy. Mantel-Haenszel chi-square and ordinal logistic regression were used to explore factors associated with the level of embarrassment. RESULTS: In total, 648 female nurses from 54 hospitals in 31 cities in China were recruited. Among these respondents, approximately 67% provided genitalia-related care at least three days per week, and about 70% of nurses felt slightly embarrassed to extremely embarrassed when providing genitalia-related care. Compared to nurses from gynecology departments, nurses from urology departments felt more embarrassed during genitalia-related care (P<0.001). Ordinal regression analysis showed that the odds of nurses from gynecology and urology departments with total empathy scores greater than 100 experiencing higher levels of embarrassment than nurses with total empathy scores less than 100 were 0.47 and 0.45, respectively. Nurses from gynecology departments with higher professional identity scores, higher education levels, more frequent genital-related care experienced less embarrassment, while nurses from the urology department with prior sexual experience experienced less embarrassment. CONCLUSIONS: Feeling embarrassed during genitalia-related nursing was common among Chinese female nurses, especially those from urology departments. Embarrassment during genitalia-related nursing was related to professional identity, empathy, educational level, genitalia-related care frequency, and sexual history. These findings highlight the importance of professional identity, empathy, and education among nurses.

No effect of 6-month supplementation with 300 mg/d docosahexaenoic acid on executive functions among healthy school-aged children: a randomized, double-blind, placebo-controlled trial.

PURPOSE: Docosahexaenoic acid (DHA) plays an essential role in brain, and its status is dependent on dietary intakes. School-aged children in rural China, who consume diets low in omega-3 polyunsaturated fatty acids, may benefit from DHA supplementation. Therefore, this trial was performed to examine the effect of 6-month DHA supplementation on executive functions (EFs) among healthy school-aged children in rural China. METHODS: A randomized, double-blind, placebo-controlled trial was conducted among 106 primary school children aged 7-12 years in rural China. Participants were randomized to receive either 300 mg/d DHA or placebo for 6 months. EFs including working memory and cognitive flexibility were evaluated at baseline, at 3 months and at 6 months, using Digit Span Backwards and Wisconsin card sorting test, respectively. Socio-demographic data were collected at baseline, and erythrocyte membrane fatty acids and serum neurotransmitters were measured at baseline and after 6-month intervention. RESULTS: Ninety-four children (88.7%) completed the study according to the protocol. Changes in erythrocyte membrane fatty acids indicated good compliance of the participants. There was no significant intervention effect on serum neurotransmitters. In two-factor ANCOVA, both groups showed a significant improvement in the Digit Span Backwards and the Wisconsin card sorting test from baseline to endpoint. However, no significant intervention effect was found on any EF scores. Linear regression analysis suggested no significant association between changes in erythrocyte DHA level with changes in any EF scores. CONCLUSIONS: Supplementation with 300 mg/d DHA for 6 months had no benefit on EFs including working memory and cognitive flexibility among healthy school-aged children. This trial was registered at clinicaltrials.gov as NCT02308930 on December 5, 2014.

A Bunyavirus-Inducible Ubiquitin Ligase Targets RNA Polymerase IV for Degradation during Viral Pathogenesis in Rice.

The ubiquitin-proteasome system (UPS) is an important post-translational regulatory mechanism that controls many cellular functions in eukaryotes. Here, we show that stable expression of P3 protein encoded by Rice grassy stunt virus (RGSV), a negative-strand RNA virus in the Bunyavirales, causes developmental abnormities similar to the disease symptoms caused by RGSV, such as dwarfing and excess tillering, in transgenic rice plants. We found that both transgenic expression of P3 and RGSV infection induce ubiquitination and UPS-dependent degradation of rice NUCLEAR RNA POLYMERASE D1a (OsNRPD1a), one of two orthologs of the largest subunit of plant-specific RNA polymerase IV (Pol IV), which is required for RNA-directed DNA methylation (RdDM). Furthermore, we identified a P3-inducible U-box type E3 ubiquitin ligase, designated as P3-inducible protein 1 (P3IP1), which interacts with OsNRPD1a and mediates its ubiquitination and UPS-dependent degradation in vitro and in vivo. Notably, both knockdown of OsNRPD1 and overexpression of P3IP1 in rice plants induced developmental phenotypes similar to RGSV disease symptomss. Taken together, our findings reveal a novel virulence mechanism whereby plant pathogens target host RNA Pol IV for UPS-dependent degradation to induce disease symptoms. Our study also identified an E3 ubiquitin ligase, which targets the RdDM compotent NRPD1 for UPS-mediated degradation in rice.

Rice Stripe Mosaic Virus-Encoded P4 Is a Weak Suppressor of Viral RNA Silencing and Is Required for Disease Symptom Development.

Viral suppressors of RNA silencing (VSRs) are a cluster of viral proteins that have evolved to counteract eukaryotic antiviral RNA silencing pathways, thereby contributing to viral pathogenicity. In this study, we revealed that the matrix protein P4 encoded by rice stripe mosaic virus (RSMV), which is an emerging cytoplasmic rhabdovirus, is a weak RNA silencing suppressor. By conducting yeast two-hybrid, bimolecular fluorescence complementation, and subcellular colocalization assays, we proved that P4 interacts with the rice endogenous suppressor of gene silencing 3 (OsSGS3). We also determined that P4 overexpression has no effect on OsSGS3 transcription. However, P4 can promote the degradation of OsSGS3 via ubiquitination and autophagy. Additionally, a potato virus X-based expression system was used to confirm that P4 enhances the development of mosaic symptoms on Nicotiana benthamiana leaves by promoting hydrogen peroxide accumulation but not cell death. To verify whether P4 is a pathogenicity factor in host plants, we generated transgenic P4-overexpressing rice plants that exhibited disease-related developmental defects including decreased plant height and excessive tillering. Our data suggest that RSMV-encoded P4 serves as a weak VSR that inhibits antiviral RNA silencing by targeting OsSGS3.

Comparative transcriptome study of hairy and hairless tea plant (Camellia sinensis) shoots.

Trichome (also referred to as 'háo' in tea) is a key feature in both tea products and tea plant (Camellia sinensis) selection breeding. Although trichomes are used as a model for studying cell differentiation and have been well studied in many plant species, the regulation of trichome formation at the molecular level is poorly understood in tea plants. In the present study, the hairy and hairless tea plant cultivars Fudingdabaicha (FDDB) and Rongchunzao (RCZ), respectively, were used to study this mechanism. We characterised tea plant trichomes as unicellular and unbranched structures. High-throughput Illumina sequencing yielded approximately 277.0 million high-quality clean reads from the FDDB and RCZ cultivars. After de novo assembly, 161,444 unigenes were generated, with an average length of 937 bp. Among these unigenes, 81,425 were annotated using public databases, and 55,201 coding sequences and 4004 transcription factors (TFs) were identified. In total, 21,599 differentially expressed genes (DEGs) were identified between RCZ and FDDB, of which 10,785 DEGs were up-regulated and 10,814 DEGs were down-regulated. Genes involved in the DNA replication pathway were significantly enriched. Furthermore, between FDDB and RCZ, DEGs related to TFs, phytohormone signals, and cellulose synthesis were identified, suggesting that certain genes involved in these pathways are crucial for trichome initiation in tea plants. Together, the results of this study provide novel data to improve our understanding of the potential molecular mechanisms of trichome formation and lay a foundation for additional trichome studies in tea plants.