RGA1 regulates grain size, rice quality and seed germination in the small and round grain mutant srg5.

BACKGROUND: Generating elite rice varieties with high yield and superior quality is the main goal of rice breeding programs. Key agronomic traits, including grain size and seed germination characteristics, affect the final yield and quality of rice. The RGA1 gene, which encodes the α-subunit of rice G-protein, plays an important role in regulating rice architecture, seed size and abiotic stress responses. However, whether RGA1 is involved in the regulation of rice quality and seed germination traits is still unclear. RESULTS: In this study, a rice mutant small and round grain 5 (srg5), was identified in an EMS-induced rice mutant library. Systematic analysis of its major agronomic traits revealed that the srg5 mutant exhibited a semi-dwarf plant height with small and round grain and reduced panicle length. Analysis of the physicochemical properties of rice showed that the difference in rice eating and cooking quality (ECQ) between the srg5 mutant and its wild-type control was small, but the appearance quality was significantly improved. Interestingly, a significant suppression of rice seed germination and shoot growth was observed in the srg5 mutant, which was mainly related to the regulation of ABA metabolism. RGA1 was identified as the candidate gene for the srg5 mutant by BSA analysis. A SNP at the splice site of the first intron disrupted the normal splicing of the RGA1 transcript precursor, resulting in a premature stop codon. Additional linkage analysis confirmed that the target gene causing the srg5 mutant phenotype was RGA1. Finally, the introduction of the RGA1 mutant allele into two indica rice varieties also resulted in small and round rice grains with less chalkiness. CONCLUSIONS: These results indicate that RGA1 is not only involved in the control of rice architecture and grain size, but also in the regulation of rice quality and seed germination. This study sheds new light on the biological functions of RGA1, thereby providing valuable information for future systematic analysis of the G-protein pathway and its potential application in rice breeding programs.

Brassinosteroids regulate rice seed germination through the BZR1-RAmy3D transcriptional module.

Seed dormancy and germination, two physiological processes unique to seed-bearing plants, are critical for plant growth and crop production. The phytohormone brassinosteroid (BR) regulates many aspects of plant growth and development, including seed germination. The molecular mechanisms underlying BR control of rice (Oryza sativa) seed germination are mostly unknown. We investigated the molecular regulatory cascade of BR in promoting rice seed germination and post-germination growth. Physiological assays indicated that blocking BR signaling, including introducing defects into the BR-insensitive 1 (BRI1) receptor or overexpressing the glycogen synthase kinase 2 (GSK2) kinase delayed seed germination and suppressed embryo growth. Our results also indicated that brassinazole-resistant 1 (BZR1) is the key downstream transcription factor that mediates BR regulation of seed germination by binding to the alpha-Amylase 3D (RAmy3D) promoter, which affects α-amylase expression and activity and the degradation of starch in the endosperm. The BZR1-RAmy3D module functions independently from the established Gibberellin MYB-alpha-amylase 1A (RAmy1A) module of the gibberellin (GA) pathway. We demonstrate that the BZR1-RAmy3D module also functions in embryo-related tissues. Moreover, RNA-sequencing (RNA-seq) analysis identified more potential BZR1-responsive genes, including those involved in starch and sucrose metabolism. Our study successfully identified the role of the BZR1-RAmy3D transcriptional module in regulating rice seed germination.

Gene editing of non-coding regulatory DNA and its application in crop improvement.

The development of the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) system has provided precise and efficient strategies to edit target genes and generate transgene-free crops. Significant progress has been made in the editing of protein-coding genes; however, studies on the editing of non-coding DNA with regulatory roles lags far behind. Non-coding regulatory DNAs, including those which can be transcribed into long non-coding RNAs (lncRNAs), and miRNAs, together with cis-regulatory elements (CREs), play crucial roles in regulating plant growth and development. Therefore, the combination of CRISPR/Cas technology and non-coding regulatory DNA has great potential to generate novel alleles that affect various agronomic traits of crops, thus providing valuable genetic resources for crop breeding. Herein, we review recent advances in the roles of non-coding regulatory DNA, attempts to edit non-coding regulatory DNA for crop improvement, and potential application of novel editing tools in modulating non-coding regulatory DNA. Finally, the existing problems, possible solutions, and future applications of gene editing of non-coding regulatory DNA in modern crop breeding practice are also discussed.

Antiproliferative and apoptotic effects of telmisartan in human glioma cells.

Glioblastoma is the most common primary central nervous system tumor in adults. Angiotensin II receptor blockers (ARBs) are broadly applied to treat hypertension. Moreover, research has revealed that ARBs have the capacity to suppress the growth of several cancer types. In this study, we assessed the effects of three ARBs with the ability to cross the blood brain barrier (telmisartan, valsartan and fimasartan) on cell proliferation in three glioblastoma multiforme (GBM) cell lines. Telmisartan markedly suppressed the proliferation, migration, and invasion of these three GBM cell lines. Microarray data analysis revealed that telmisartan regulates DNA replication, mismatch repair, and the cell cycle pathway in GBM cells. Furthermore, telmisartan induced G0/G1 phase arrest and apoptosis. The bioinformatic analysis and western blotting results provide evidence that SOX9 is a downstream target of telmisartan. Telmisartan also suppressed tumor growth in vivo in an orthotopic transplant mouse model. Therefore, telmisartan is a potential treatment for human GBM.

[Association between dietary patterns and abdominal obesity among the adult women in Zhejiang Province].

OBJECTIVE: To explore the dietary patterns and its relationship with abdominal obesity among the adult women in Zhejiang Province. METHODS: We recruited 2915 female residents aged 18 and older by using multi-stage stratified cluster sampling method; We conducted questionnaires survey and physical examinations to understand the basic information and nutritional status of the residents. Dietary patterns were extracted with factor analysis, and the multivariate Logistic regression model was used to analyze the association between dietary patterns and abdominal obesity among the participants. RESULTS: The prevalence rate of abdominal obesity and general obesity among the subjects were 33.07% and 9.23%, respectively. The highest rate of abdominal obesity was among the female who were above 65 years old, living in rural areas with low education. On the other hand, the lowest rate of abdominal obesity was among females with higher education(P<0.0001). Four dietary patterns identified with factor analysis were grain pattern, nut-legume pattern, milk-eggs pattern and modern pattern, accounting for 38.5% of total variance. After adjusting confounding factors as age, education level, occupation, marital status, urban/rural and exercise status, the participants with the fourth quartile(Q4) score of nut-legume dietary pattern had a low risk of abdominal obesity(OR=0.720, 95% CI 0.571-0.908, P=0.0012) compared to those with the quartile(Q1) first score. CONCLUSION: The nut-legume dietary pattern is negatively associated with abdominal obesity in female adults.

Improving rice eating and cooking quality by coordinated expression of the major starch synthesis-related genes, SSII and Wx, in endosperm.

KEY MESSAGE: Coordinated regulation of amylose and amylopectin synthesis via manipulation of SSII-2, SSII-3 and Wx expression in endosperm can improve rice eating and cooking quality. With increasing rice consumption worldwide, many researchers are working to increase the yield and improve grain quality, especially eating and cooking quality (ECQ). The rice ECQ is mainly controlled by the expression of starch synthesis-related genes (SSRGs) in endosperm. Although the Wx and SSII-3/SSIIa/ALK genes, two major SSRGs, have been manipulated to improve rice ECQ via various breeding approaches, new methods to further improve ECQ are desired. In our previous study, we enhanced rice ECQ by knocking down SSII-2 expression in the japonica Nipponbare cultivar (carrying the Wxb allele) via RNA interference. Herein, the SSII-2 RNAi was introduced into two Nipponbare-derived near-isogenic lines (NILs), Nip(Wxa) and Nip(wx), carrying Wxa and wx alleles respond for high and no amylose levels, respectively. Analysis of physicochemical properties revealed that the improved grain quality of SSII-2 RNAi transgenic lines was achieved by coordinated downregulating the expression of SSII-2, SSII-3 and Wx. To further confirm this conclusion, we generated ssii-2, ssii-3 and ssii-2ssii-3 mutants via CRISPR/Cas9 technique. The amylopectin structure of the resulting ssii-2sii-3 mutants was similar to that in SSII-2 RNAi transgenic lines, and the absence of SSII-2 decreased the amylose content, gelatinisation temperature and rapid visco-analyser profile, indicating essential roles for SSII-2 in the regulation of amylopectin biosynthesis and amylose content in rice endosperm. The effect of SSII-2 was seen only when the activity of SSII-3 was very low or lacking. Our study provides novel approaches and valuable germplasm resources for improving ECQ via plant breeding.

Glucan, Water-Dikinase 1 (GWD1), an ideal biotechnological target for potential improving yield and quality in rice.

The source-sink relationship determines the overall agronomic performance of rice. Cloning and characterizing key genes involved in the regulation of source and sink dynamics is imperative for improving rice yield. However, few source genes with potential application in rice have been identified. Glucan, Water-Dikinase 1 (GWD1) is an essential enzyme that plays a pivotal role in the first step of transitory starch degradation in source tissues. In the present study, we successfully generated gwd1 weak mutants by promoter editing using CRISPR/Cas9 system, and also leaf-dominant overexpression lines of GWD1 driven by Osl2 promoter. Analysis of the gwd1 plants indicated that promoter editing mediated down-regulation of GWD1 caused no observable effects on rice growth and development, but only mildly modified its grain transparency and seed germination. However, the transgenic pOsl2::GWD1 overexpression lines showed improvements in multiple key traits, including rice yield, grain shape, rice quality, seed germination and stress tolerance. Therefore, our study shows that GWD1 is not only involved in transitory starch degradation in source tissues, but also plays key roles in the seeds, which is a sink tissue. In conclusion, we find that GWD1 is an ideal biotechnological target with promising potential for the breeding of elite rice cultivars via genetic engineering.

The Current Status of Insufficient Sleep Duration and Its Influencing Factors Among Children and Adolescents: A Household Based Cross-Sectional Study in Zhejiang Province, China.

PURPOSE: Insufficient sleep duration is common among children and adolescents worldwide, and the decline of sleep duration during the recent years is troubling. This study aimed to learn the status of insufficient sleep duration and to explore its influencing factors among children and adolescents in Zhejiang Province, China. DESIGN AND METHODS: A stratified sampling technique was employed in the present cross-sectional study. Demographic characteristics, sports time as well as sedentary behavior were investigated. RESULTS: A greater proportion of children than adolescents reported insufficient sleep duration (36.4% versus 19.2%, p = 0.001). For children, insufficient sleep duration was associated with age (OR = 1.290, 95%CI: 1.069-1.557), watching movies or TV shows with smartphones after school (OR = 3.098, 95%CI: 1.293-7.420), surf the internet after school (OR = 0.113, 95%CI: 0.013-0.969), walk to school (OR = 0.289, 95%CI: 0.105-0.793). For adolescents, insufficient sleep duration was associated with watch TV after school (OR = 0.379, 95%CI: 0.148-0.970), watching movies or TV shows with smartphones after school (OR = 4.744, 95%CI: 1.799-12.507), do homework after school (OR = 0.265, 95%CI: 0.086-0.813). CONCLUSIONS: An unhealthy sedentary screen lifestyle profile may have influence on insufficient sleep duration. Urgent actions are needed to improve sleep duration among children and adolescents.

Diosmin Inhibits Glioblastoma Growth through Inhibition of Autophagic Flux.

Diosmin, a natural flavone glycoside acquired through dehydrogenation of the analogous flavanone glycoside hesperidin, is plentiful in many citrus fruits. Glioblastoma multiforme (GBM) is the most malignant primary brain tumor; the average survival time of GBM patients is less than 18 months after standard treatment. The present study demonstrated that diosmin, which is able to cross the blood-brain barrier, inhibited GBM cell growth in vitro and in vivo. Diosmin also impeded migration and invasion by GBM8401and LN229 GBM cells by suppressing epithelial-mesenchymal transition, as indicated by increased expression of E-cadherin and decreased expression of Snail and Twist. Diosmin also suppressed autophagic flux, as indicated by increased expression of LC3-II and p62, and induced cell cycle arrest at G1 phase. Importantly, diosmin did not exert serious cytotoxic effects toward control SVG-p12 astrocytes, though it did reduce astrocyte viability at high concentrations. These findings provide potentially helpful support to the development of new therapies for the treatment of GBM.

GRPEL2 Knockdown Exerts Redox Regulation in Glioblastoma.

Malignant brain tumors are responsible for catastrophic morbidity and mortality globally. Among them, glioblastoma multiforme (GBM) bears the worst prognosis. The GrpE-like 2 homolog (GRPEL2) plays a crucial role in regulating mitochondrial protein import and redox homeostasis. However, the role of GRPEL2 in human glioblastoma has yet to be clarified. In this study, we investigated the function of GRPEL2 in glioma. Based on bioinformatics analyses from the Cancer Gene Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), we inferred that GRPEL2 expression positively correlates with WHO tumor grade (p < 0.001), IDH mutation status (p < 0.001), oligodendroglial differentiation (p < 0.001), and overall survival (p < 0.001) in glioma datasets. Functional validation in LN229 and GBM8401 GBM cells showed that GRPEL2 knockdown efficiently inhibited cellular proliferation. Moreover, GRPEL2 suppression induced cell cycle arrest at the sub-G1 phase. Furthermore, GRPEL2 silencing decreased intracellular reactive oxygen species (ROS) without impending mitochondria membrane potential. The cellular oxidative respiration measured with a Seahorse XFp analyzer exhibited a reduction of the oxygen consumption rate (OCR) in GBM cells by siGRPEL2, which subsequently enhanced autophagy and senescence in glioblastoma cells. Taken together, GRPEL2 is a novel redox regulator of mitochondria bioenergetics and a potential target for treating GBM in the future.

SNAP reverses temozolomide resistance in human glioblastoma multiforme cells through down-regulation of MGMT.

Glioblastoma multiforme (GBM) is the most frequently occurring and gravest primary tumor of the CNS in adults. The development of chemoresistance to temozolomide (TMZ), the first-line chemotherapy for GBM, is an important factor contributing to poor treatment outcomes. Down-regulation of O-6-methylguanine-DNA methyltransferase (MGMT) expression in GBM cells is an attractive strategy for overcoming TMZ resistance and improving outcomes. This study revealed that the nitric oxide (NO) donor S-nitroso-N-acetylpenicillamine (SNAP) exerts antitumorigenic effects on TMZ-sensitive and TMZ-resistant (TMZ-R) glioma cells. Pretreatment with SNAP not only induced apoptosis, mitochondrial dysfunction, and hypoxia-inducing factor 1, but also resensitized TMZ-R GBM cells to TMZ through down-regulation of MGMT expression. SNAP acted principally through post-translational modification of p53, phosphorylated N-myc downstream regulated gene 1, and MGMT protein stability in TMZ-R GBM cells. Additionally, when applied together, SNAP and TMZ enhanced the inhibition of tumor growth in vitro and in vivo. This study sheds new light on a potential strategy to overcome TMZ resistance in GBM and thus possesses the potential for prolonging survival of patients with GBM.-Tsai, C.-K., Huang, L.-C., Wu, Y.-P., Kan, I.-Y., Hueng, D.-Y. SNAP reverses temozolomide resistance in human glioblastoma multiforme cells through down-regulation of MGMT.

Adult breakfast habits and nutritional status: A household based cross-sectional study in Zhejiang Province, China.

BACKGROUND AND OBJECTIVES: Although Dietary Guidelines for Chinese Residents offers advice for breakfast in China, current breakfast consumption habits are unclear. The aim of this study was to describe the frequency of breakfast consumption among adults and explore its associations with daily food consumption and daily nutrient intake. METHODS AND STUDY DESIGN: A stratified cluster sampling technique was employed in this cross-sectional study. Demographic characteristics, breakfast consumption, and daily food and nutrient intake were investigated. RESULTS: Dietary data were collected for 3251 adult residents through interviews. We determined that residents with an annual income of

CKAP2L Knockdown Exerts Antitumor Effects by Increasing miR-4496 in Glioblastoma Cell Lines.

Despite advances in the diagnosis and treatment of the central nervous system malignancy glioma, overall survival remains poor. Cytoskeleton-associated protein 2-like (CKAP2L), which plays key roles in neural progenitor cell division, has also been linked to poor prognosis in lung cancer. In the present study, we investigated the role of CKAP2L in glioma. From bioinformatics analyses of datasets from The Cancer Gene Atlas and the Chinese Glioma Genome Atlas, we found that CKAP2L expression correlates with tumor grade and overall survival. Gene set enrichment analysis (GSEA) showed that MITOTIC_SPINDLE, G2M_CHECKPOINT, and E2F_TARGETS are crucially enriched phenotypes associated with high CKAP2L expression. Using U87MG, U118MG, and LNZ308 human glioma cells, we confirmed that CKAP2L knockdown with siCKAP2L inhibits glioma cell proliferation, migration, invasion, and epithelial-mesenchymal transition. Interestingly, CKAP2L knockdown also induced cell cycle arrest at G2/M phase, which is consistent with the GSEA finding. Finally, we observed that CKAP2L knockdown led to significant increases in miR-4496. Treating cells with exogenous miR-4496 mimicked the effect of CKAP2L knockdown, and the effects of CKAP2L knockdown could be suppressed by miR-4496 inhibition. These findings suggest that CKAP2L is a vital regulator of miR-4496 activity and that CKAP2L is a potentially useful prognostic marker in glioma.

[Dietary microelements intakes status of elderly people from different areas in Zhejiang Province in 2010-2012].

OBJECTIVE: To assess the situations of dietary microelements intakes among elderly people in different areas of Zhejiang Province, and analyze the food sources of microelements. METHODS: Data were obtained from the 2010-2012 Chinese National Nutrition and Health Survey in Zhejiang Province. 832 elderly people( 434 male, 398 female, age was( 68. 02±6. 68) years) were from large cities, small-medium cities and rural counties. The 24-hour dietary recall method for 3 consecutive days was used to collect food intake information to analyze dietary intakes among elderly people of Fe, Mn, Cu, Zn, Se and their sources in different areas. RESULTS: The elderly people'intakes of Mn was 4. 86( 3. 44, 6. 45) mg/d, Cu was 1. 69( 1. 13, 2. 27) mg/d, and Se was36. 85( 21. 32, 54. 21) µg/d, those were generally low. The intake of Zn was 10. 15( 7. 94, 12. 62) mg/d in males, was low as well. Intakes of Mn, Cu, Se were significantly different in different areas, small-medium cities had highest intakes of Mn( 5. 87 mg/d)and Cu( 1. 88 mg/d), large cities had a highest intake of Se( 45. 47 µg/d). The food sources of microelements were slightly variant. Rural counties had less fungi and algae sources of Fe and Mn than large cities and small-medium cities. The Cu from vegetables were higher than dried legumes and the Se from poultry and poultry products were higher than eggs in rural counties. The Zn from fish, shellfish and mollusk were higher than dried legumes in large cities. CONCLUSION: The insufficient status of microelements intakes among elderly people in Zhejiang Province was serious, and there were differences among elderly people from different areas in intakes and food sources of microelements.

[A cohort study of serum leptin level in Zhejiang Province middle and old age residents on blood pressure and hypertension from 2014 to 2017].

OBJECTIVE: To investigate the association between serum leptin concentration with blood pressure and hypertension in different gender. METHODS: A total of 343 non-hypertensive residents aged 30 to 65 were randomly selected from Zhejiang Province in 2014. Parameters including height, weight, waist, blood pressure, serum lipid and serum leptin concentration of participants were measured and determined by physical examination and laboratory detection in 2014 and 2017. The differences in serum leptin levels between the new hypertensive and non-hypertensive people were compared after three years. The relationship between serum leptin levels and hypertension was analyzed by Logistic regression. RESULTS: In 2014 and 2017, concentration of serum leptin in females was significantly higher than that in males, and the blood pressure in males was higher than that in females. At the end of follow-up, 26 new cases of hypertension were found. Serum leptin levels were higher in the female hypertensive group than in the non-hypertensive group(P=0. 0289), whereas there was no statistical difference in males(P>0. 01). Regardless of gender, serum leptin was significantly positively correlated with body mass index(BMI) and waist-to-height ratio(WHtR)(P<0. 0001), and also had correlation with blood pressure. However, after adjusting for BMI or WHtR, the correlation between serum leptin and blood pressure disappeared(P>0. 01). Logistic regression also showed the similar result. CONCLUSION: Serum leptin is not directly related to blood pressure and hypertension, and the positive correlation between them may be explained largely by BMI and WHtR.

[Comparative study on burden of diet-related chronic diseases in China between 1990 and 2016].

OBJECTIVE: To compare the disease burden of diet-related chronic diseases in China between 1990 and 2016. METHODS: Based on the Global Burden of Disease 2016, we extracted the absolute number of deaths and disability-adjusted life years( DALYs) of diet-related chronic diseases and the corresponding age-standardized rates in China in 1990 and 2016. Subgroup analyses were conducted in various gender, dietary risk factors and categories of diseases. RESULTS: In 2016, the total number and rate of diet-related NCDs deaths was 2493 thousand and 182. 4/100000. The corresponding number and rate of DALYs was 54995 thousand person-year and 4023. 0/100000. Compared with those in 1990, the rates of death and DALYs in 2016 increased by19. 5% and 9. 3%, respectively. Both death rate and DALYs rate increased with the age. Death rate( 214. 6/100000) and DALYs rate( 4961. 1/100000) in males were 1. 4-fold and 1. 6-fold as that in females( 148. 2/100000 and 3028. 2/100000, respectively). High intake in sodium, low intake in whole grains and low intake in fruits were the three leading dietary risk factors and cardiovascular diseases accounted for 88. 1% DALYs in all chronic diseases. CONCLUSION: The burden of diet-related chronic diseases in China is severe and it continues increasing over years.

iTRAQ-Based Analysis of Proteins Co-Regulated by Brassinosteroids and Gibberellins in Rice Embryos during Seed Germination.

Seed germination, a pivotal process in higher plants, is precisely regulated by various external and internal stimuli, including brassinosteroid (BR) and gibberellin (GA) phytohormones. The molecular mechanisms of crosstalk between BRs and GAs in regulating plant growth are well established. However, whether BRs interact with GAs to coordinate seed germination remains unknown, as do their common downstream targets. In the present study, 45 differentially expressed proteins responding to both BR and GA deficiency were identified using isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis during seed germination. The results indicate that crosstalk between BRs and GAs participates in seed germination, at least in part, by modulating the same set of responsive proteins. Moreover, most targets exhibited concordant changes in response to BR and GA deficiency, and gene ontology (GO) indicated that most possess catalytic activity and are involved in various metabolic processes. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis was used to construct a regulatory network of downstream proteins mediating BR- and GA-regulated seed germination. The mutation of GRP, one representative target, notably suppressed seed germination. Our findings not only provide critical clues for validating BR⁻GA crosstalk during rice seed germination, but also help to optimise molecular regulatory networks.

[Analysis on the dietary survey results of 60 years old and over population in different economic area of Zhejiang Province in 2010-2012].

OBJECTIVE: To analyze the dietary energy and pattern of the elderly in different economic area of Zhejiang Province. METHODS: Data was from Chinese National Nutrition and Health Survey from 2010 to 2012 in Zhejiang Province. The 24 h dietary recall method for 3 consecutive days was used in dietary survey and weighing method was used for condiment intake. RESULTS: There were 793 elderly( age≥60 years) attended in the survey. The intake of beans and nuts, fish and shrimps, eggs, cereals and vegetables respectively were 30. 26, 54. 07, 20. 56, 300. 27 and 277. 78 g. The livestock and poultry meat( 92. 69 g), salt( 9. 14 g), and cooking oil( 33. 60 g) intake was higher than that recommended. Dietary pattern of the urban was better than that of the rural areas. The proportion of energy providing in protein and fat was 14. 17% and 34. 46%, respectively. The proportion of protein from grain and animal food was 31. 51% and 37. 48%. The proportion of fat from plant food was 66. 44%. CONCLUSION: The major problems areirregular diet, imbalanced diet, and unreasonable food sources of protein in Zhejiang province. Because the level of each localities economic development is different, each region's dietary problems are different. We need to research in different areas to analyze specific issues of nutrition interventions, and improving the elders' life quality.