A soybean bZIP transcription factor is involved in submergence resistance.

Although the functions of basic leucine zipper (bZIP) family transcription factors in the regulation of various abiotic stresses are beginning to be unveiled, the precise roles of bZIP proteins in plants coping with submergence stress remain unclear. Here we identified a bZIP gene GmbZIP71-4 from soybean, which localized in the nucleus. The GmbZIP71-4 over-expressed tabocco line showed reduced submergence resistance due to the decreased abscisic acid (ABA) content. GO and KEGG pathway analysis based on chromatin immunoprecipitation assay sequencing (ChIP-seq) indicated that the differences expressed genes between submergence treatment and control groups were specially enriched in plant hormone signal transduction items, especially those in response to ABA. Electrophoretic mobility shift assays (EMSA) demonstrated that GmbZIP71-4 bound to the promoter of GmABF2 gene, which is consistent with the ChIP-qPCR results. GmbZIP71-4 function as a negative regulator of soybean in responding to submergence stress through manipulating ABA signaling pathway. This findings will set a solid foundation for the understanding of submergence resistance in plants.

Single-cell RNA sequencing reveals roles of unique retinal microglia types in early diabetic retinopathy.

BACKGROUND: The pathophysiological mechanisms of diabetic retinopathy (DR), a blinding disease, are intricate. DR was thought to be a microvascular disease previously. However, growing studies have indicated that the retinal microglia-induced inflammation precedes microangiopathy. The binary concept of microglial M1/M2 polarization paradigms during inflammatory activation has been debated. In this study, we confirmed microglia had the most significant changes in early DR using single-cell RNA sequencing. METHODS: A total of five retinal specimens were collected from donor SD rats. Changes in various cells of the retina at the early stage of DR were analyzed using single-cell sequencing technology. RESULTS: We defined three new microglial subtypes at cellular level, including two M1 types (Egr2+ M1 and Egr2- M1) and one M2 type. We also revealed the anatomical location between these subtypes, the dynamic changes of polarization phenotypes, and the possible activation sequence and mutual activation regulatory mechanism of different cells. Furthermore, we constructed an inflammatory network involving microglia, blood-derived macrophages and other retinal nonneuronal cells. The targeted study of new disease-specific microglial subtypes can shorten the time for drug screening and clinical application, which provided insight for the early control and reversal of DR. CONCLUSIONS: We found that microglia show the most obvious differential expression changes in early DR and reveal the changes in microglia in a high-glucose microenvironment at the single-cell level. Our comprehensive analysis will help achieve early reversal and control the occurrence and progression of DR.

Denticleless E3 ubiquitin protein ligase (DTL) maintains the proliferation and differentiation of epidermis and hair follicles during skin development.

BACKGROUND: A precise balance between the proliferation and differentiation of epidermal progenitors is required to achieve the barrier function during the development of epidermis. During the entire process of skin development, the newly formed basal layer cells divide, differentiate, and migrate outward to the surface of the skin, which is tightly regulated by a series of events related to cell cycle progression. The CRL4DTL complex (Cullin 4 RING ligase, in association with the substrate receptor DTL) has long emerged as a master regulator in various cellular processes, which mediates the degradation of key cell cycle proteins. However, the roles of DTL in regulating epidermal morphogenesis during skin development remain unclear. RESULTS: We showed that DTL deficiency in epidermal progenitor cells leads to defects in epidermal stratification and loss of hair follicles accompanied by reduced epidermal progenitor cells and disturbed cell cycle progression during skin development. Transcriptome analysis revealed that p53 pathway is activated in DTL-depleted epidermal progenitor cells. The apoptosis of epidermal cells showed in DTL deficiency mice is rescued by the absence of p53, but the proliferation and differentiation defects were p53-independent. CONCLUSION: Our findings indicate that DTL plays a vital role in epidermal malformation during skin development.

Analysis of the epidemiological burden of age-related macular degeneration in China based on the data of global burden of disease. / 基于GBDæ•°æ®åº“ä¸­å›½å¹´é¾„ç›¸å ³æ€§é»„æ–‘å˜æ€§æµè¡Œç— å­¦è´Ÿæ‹ åˆ†æž.

OBJECTIVES: Age-related macular degeneration (AMD) is one of the 3 major eye diseases recognized by WHO to prevent blindness, and which is the main cause of irreversible visual impairment in the elderly. This study aims to analyze the disease epidemiological burden, and provide a theoretical foundation for the prevention and control of AMD in China based on the data in global burden of disease (GBD) 2019. METHODS: The prevalent cases/prevalence, disability-adjusted life year (DALYs)/DALY rate of AMD and socio-demographic index (SDI) for global and China were searched from the GBD 2019 database to analyze the epidemiological trend, age-period-gender trend of AMD in China from 1990 to 2019, and to evaluate the relations between the prevalence and SDI. RESULTS: In 2019, the prevalence of AMD in China was at a high level in the world, and the number of prevalent cases were 1.93 times of that in 1990. The prevalence and DALY rates continued to rise. The age trend of AMD in China was high at the middle of the age stages and low at the two ends, and which was higher in the female than in the male. With the increase of SDI, the prevalence of AMD was increased linearly. CONCLUSIONS: The disease burden of AMD in China is increased significantly and is positively correlated with the social development from 1990 to 2019. It is of great significance to study the relationship between epidemilolgical data of AMD and social development level for diagnosis treatment and policy of AMD.

Optimal exercise combinations for patients with diabetes. / ç³–å°¿ç— æ‚£è€ çš„æœ€ä½³è¿åŠ¨ç»„åˆæ¨¡å¼.

OBJECTIVES: Exercise is a key way to control the blood glucose, which can improve blood lipids and blood pressure, reduce glycated hemoglobin levels, and increase insulin sensitivity. It is of great significance for maintaining blood glucose homeostasis. This study aims to explore the optimal exercise combinations suitable for diabetic patients, and to provide scientific and effective personalized exercise guidance for diabetic patients. METHODS: The physical examination data and questionnaire results of were collected from the Third Xiangya Hospital of Central South University. The study was involved in 3 867 diabetes from January, 2020 to December, 2021. The basic information and living habits were obtained through questionnaires. The fasting blood samples were collected to measure the levels of total cholesterol (TCh), triglyceride, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and fasting blood glucose. Physical measurements included height, weight, waistline, hipline, and blood pressure. Categorical variables were tested using chi-square tests, and continuous variables were tested using non-parametric Kruskal-Wallis test. Multiple linear regression model was used to analyze the relationship between exercise behaviors (frequency, duration, and intensity), years of exercise, diet habits, medicine, and fasting blood glucose. The relationship between different exercise behaviors and various indexes (BMI, blood pressure, blood lipids, blood glucose) was analyzed by multiple rising sun rose plots. The polar thermal diagram showed the relationship of exercise behaviors with the best expected effect. RESULTS: Non-parametric Kruskal-Wallis test showed that there were significant differences in other indexes except LDL-C (P=0.681) among groups with different exercise behaviors (all P<0.01). The multiple rising sun rose plots showed that diabetic patients who did moderate to high intensity exercise more than 5 times a week for more than 30 min each time had a healthier BMI, blood lipid levels, blood glucose levels, and blood pressure. Polar thermal diagram showed that low-intensity exercise once or twice a week requires 30 to 60 min or more of exercise to achieve the desired results. Multiple linear regression analysis showed that frequency (ß=-0.208, 95% CI -0.356 to -0.059, P=0.006), duration (ß=-0.227, 95% CI -0.387 to -0.066, P=0.006), intensity of exercise (ß=-0.110, 95% CI -0.218 to -0.002, P=0.046), diet habits (ß=0.462, 95% CI 0.295 to 0.556, P<0.001) and medicine (ß=-0.520, 95% CI -0.720 to -0.312, P<0.001) were correlated with fasting blood glucose. CONCLUSIONS: Moderate to high intensity exercise for more than 30 min and 5 times a week is the most beneficial combination for diabetes. Low exercise intensity should be combined with higher exercise frequency and longer exercise time to achieve the desired effect. With the increase of exercise intensity, the relationship between low exercise frequency and long exercise time is weakened.

The grain yield regulator NOG1 plays a dual role in latitudinal adaptation and cold tolerance during rice domestication.

Rice originated in tropical and subtropical regions and is distributed worldwide. Low temperature is one of the most critical abiotic stresses affecting grain yield and geographical distribution of rice. It is vital to elucidate the molecular mechanism of chilling tolerance in rice for ensuring cereals production. Previously we isolated the domestication-related gene NOG1 which affects rice grain number and yield. In this study, we specified that rice varieties harboring high-yielding NOG1 allele are more distributed in low-latitude regions. Additionally, we observed NOG1 influences the chilling tolerance of rice. Through genome-wide transcriptional analysis after cold treatment at 10°C, there were 717 differentially expressed genes (DEGs) in nog1 near-isogenic lines compared with the control Guichao 2, including 432 up-regulated DEGs and 284 down-regulated DEGs. Gene ontology annotations and KEGG enrichment analysis of DEGs showed that various biological processes and signaling pathways were related to cold stress, such as lipid metabolism and genetic information processing. These results provide new insights into the mechanism of chilling tolerance in rice and the molecular basis of environmental adaptation during rice domestication.

The frequency of early age-related macular degeneration and its relationship with dietary pattern in Hunan, China: a cross-sectional study.

PURPOSE: To estimate the frequency of age-related macular degeneration (AMD) among people who underwent health examination in Hunan, China and to determine the relationship between dietary pattern and the risk of AMD. METHODS: The Questionnaire was used to collect dietary data from 56,775 study participants of ≥ 50 years old who underwent health examination at the Department of Health Management, the Third Xiangya Hospital of Central South University between January 2017 and December 2019. The diagnosis of AMD was based on the results of color fundus photography (CFP), spectral-domain optical coherence tomography (OCT) and multispectral imaging (MSI). After excluding participants with incomplete records or other ocular disease that may affect the results of fundus examination, a total of 43,672 study participants were included. The univariate and multivariate logistic regression analyses were used to determine the relationship between dietary pattern and the frequency of AMD. RESULTS: Among the 43,672 study participants, 1080 (2.5%) had early AMD: the frequencies were 2.6% (n = 674) in men and 2.3% (n = 406) in women; the frequencies were 1.0% (n = 289), 3.6% (n = 401), 9.1% (n = 390) in 50-59, 60-69, ≥ 70 years old, respectively. And the age-standard frequency was 6.6% over the 60 years old in Hunan China. The high-salt intake increased the risk of early AMD [odds ratio (OR) = 1.61, 95% confidence interval (CI) = 1.54-1.68], whereas the intake of meat decreased the risk (OR = 0.90, 95% CI = 0.81-0.99). CONCLUSION: In Hunan China, there was a high frequency of early AMD detected through health examination over the 60 years old. And high-salt intake increases the risk of early AMD, whereas intake of meat decreases the risk. Modulating the dietary pattern and reducing the salt intake as an AMD prevention strategy warrant further study.

Small EPIDERMAL PATTERNING FACTOR-LIKE2 peptides regulate awn development in rice.

The EPIDERMAL PATTERNING FACTOR (EPF) and EPF-LIKE (EPFL) family of small secreted peptides act to regulate many aspects of plant growth and development; however, their functions are not widely characterized in rice (Oryza sativa). Here, we used clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) technology to individually knockout each of 11 EPF/EPFL genes in the rice cultivar Kasalath. Loss of function of most OsEPF/EPFL genes generated no obvious phenotype alteration, while disruption of OsEPFL2 in Kasalath caused a short or no awn phenotype and reduced grain size. OsEPFL2 is strongly expressed in the young panicle, consistent with a role in regulating awn and grain development. Haplotype analysis indicated that OsEPFL2 can be classified into six major haplotypes. Nucleotide diversity and genetic differentiation analyses suggested that OsEPFL2 was positively selected during the domestication of rice. Our work to systematically investigate the function of EPF/EPFL peptides demonstrates that different members of the same gene family have been independently selected for their ability to regulate a similar biological function and provides perspective on rice domestication.

Comparison Between Statistical Model and Machine Learning Methods for Predicting the Risk of Renal Function Decline Using Routine Clinical Data in Health Screening.

Purpose: Using machine learning method to predict and judge unknown data offers opportunity to improve accuracy by exploring complex interactions between risk factors. Therefore, we evaluate the performance of machine learning (ML) algorithms and to compare them with logistic regression for predicting the risk of renal function decline (RFD) using routine clinical data. Patients and Methods: This retrospective cohort study includes datasets from 2166 subjects, aged 35-74 years old, provided by an adult health screening follow-up program between 2010 and 2020. Seven different ML models were considered - random forest, gradient boosting, multilayer perceptron, support vector machine, K-nearest neighbors, adaptive boosting, and decision tree - and were compared with standard logistic regression. There were 24 independent variables, and the baseline estimate glomerular filtration rate (eGFR) was used as the predictive variable. Results: A total of 2166 participants (mean age 49.2±11.2 years old, 63.3% males) were enrolled and randomly divided into a training set (n=1732) and a test set (n=434). The area under receiver operating characteristic curve (AUROC) for detecting RFD corresponding to the different models were above 0.85 during the training phase. The gradient boosting algorithms exhibited the best average prediction accuracy (AUROC: 0.914) among all algorithms validated in this study. Based on AUROC, the ML algorithms improved the RFD prediction performance, compared to logistic regression model (AUROC:0.882), except the K-nearest neighbors and decision tree algorithms (AUROC:0.854 and 0.824, respectively). However, the improvement differences with logistic regression were small (less than 4%) and nonsignificant. Conclusion: Our results indicate that the proposed health screening dataset-based RFD prediction model using ML algorithms is readily applicable, produces validated results. But logistic regression yields as good performance as ML models to predict the risk of RFD with simple clinical predictors.

Comparative transcriptome and weighted correlation network analyses reveal candidate genes involved in chlorogenic acid biosynthesis in sweet potato.

Chlorogenic acids (CGAs) are important secondary metabolites produced in sweet potato. However, the mechanisms of their biosynthesis and regulation remain unclear. To identify potential genes involved in CGA biosynthesis, analysis of the dynamic changes in CGA components and RNA sequencing were performed on young leaves (YL), mature leaves (ML), young stems (YS), mature stems (MS) and storage roots (SR). Accordingly, we found that the accumulation of six CGA components varied among the different tissues and developmental stages, with YS and YL recording the highest levels, while SR exhibited low levels. Moreover, the transcriptome analysis yielded 59,287 unigenes, 3,767 of which were related to secondary-metabolite pathways. The differentially expressed genes (DEGs) were identified based on CGA content levels by comparing the different samples, including ML vs. YL, MS vs. YS, SR vs. YL and SR vs. YS. A total of 501 common DEGs were identified, and these were mainly implicated in the secondary metabolites biosynthesis. Additionally, eight co-expressed gene modules were identified following weighted gene co-expression network analysis, while genes in darkgrey module were highly associated with CGA accumulation. Darkgrey module analysis revealed that 12 unigenes encoding crucial enzymes (PAL, 4CL, C4H, C3H and HCT/HQT) and 42 unigenes encoding transcription factors (MYB, bHLH, WD40, WRKY, ERF, MADS, GARS, bZIP and zinc finger protein) had similar expression patterns with change trends of CGAs, suggesting their potential roles in CGA metabolism. Our findings provide new insights into the biosynthesis and regulatory mechanisms of CGA pathway, and will inform future efforts to build a genetically improve sweet potato through the breeding of high CGA content varieties.

Receptor-selective interleukin-4 mutein attenuates laser-induced choroidal neovascularization through the regulation of macrophage polarization in mice.

Macrophage polarization has been recognized as an important inflammatory regulator in choroidal neovascularization (CNV). However, the mechanisms regulating macrophage activation and polarization, as well as their effects on angiogenesis and CNV, have not yet been elucidated. IL-4 is implicated in macrophage activation and exerts different functions in various diseases through several receptors. In the current study, the effect of IL-4 muteins on CNV was investigated in vivo. CNV was induced by laser coagulation in wild type mice. IL-4 muteins were recombined into adenoviruses and injected into mice via the tail vein. To evaluate CNV, fluorescein fundus angiography and optical coherence tomography were performed on day 7 after coagulation. Quantitative PCR, western blotting and immunofluorescence staining were used to assess the levels of inflammatory markers. AdIL-4/Q116E, an adenovirus-expressed recombinant IL-4 mutein, selectively activated macrophages, alleviated laser-induced CNV in mice with reduced expression of M2 macrophages and increased the expression of M1 macrophages. Furthermore, the expression of monocyte to macrophage differentiation-associated and delta-like 4 (Dll4) in CNV lesions was upregulated. Employing AdIL-4/Q116E, a IL-4RI-selective mutein, may serve as a new strategy for CNV therapy. Moreover, the results indicated that Dll4 signaling served an important role in the regulation of macrophage polarization.

Genome-wide characterization and expression analysis of soybean trihelix gene family.

Trihelix transcription factors play multiple roles in plant growth, development and various stress responses. In this study, we identified 71 trihelix family genes in the soybean genome. These trihelix genes were located at 19 out of 20 soybean chromosomes unevenly and were classified into six distinct subfamilies: GT-1, GT-2, GTγ, SIP1, SH4 and GTδ. The gene structure and conserved functional domain of these trihelix genes were similar in the same subfamily but diverged between different subfamilies. Thirteen segmental duplicated gene pairs were identified and all of them experienced a strong purifying selective pressure during evolution. Various stress-responsive cis-elements presented in the promoters of soybean trihelix genes, suggesting that the trihelix genes might respond to the environmental stresses in soybean. The expression analysis suggests that trihelix genes are involved in diverse functions during soybean development, flood or salinity tolerance, and plant immunity. Our results provide genomic information of the soybean trihelix genes and a basis for further characterizing their roles in response to environmental stresses.

Identification of Genes/Proteins Related to Submergence Tolerance by Transcriptome and Proteome Analyses in Soybean.

Flooding can lead to yield reduction of soybean. Therefore, identification of flooding tolerance genes has great significance in production practice. In this study, Qihuang 34, a highly-resistant variety to flooding stress, was selected for submergence treatments. Transcriptome and proteome analyses were conducted, by which twenty-two up-regulated differentially expressed genes (DEGs)/differentially expressed proteins (DEPs) associated with five KEGG pathways were isolated. The number of the DEGs/DEPs enriched in glycolysis/gluconeogenesis was the highest. Four of these genes were confirmed by RT-qPCR, suggesting that glycolysis/gluconeogenesis may be activated to generate energy for plant survival under anaerobic conditions. Thirty-eight down-regulated DEGs/DEPs associated with six KEGG pathways were identified under submergence stress. Eight DEGs/DEPs enriched in phenylpropanoid biosynthesis were assigned to peroxidase, which catalyzes the conversion of coumaryl alcohol to hydroxy-phenyl lignin in the final step of lignin biosynthesis. Three of these genes were confirmed by RT-qPCR. The decreased expression of these genes led to the inhibition of lignin biosynthesis, which may be the cause of plant softening under submergence stress for a long period of time. This study revealed a number of up-/down-regulated pathways and the corresponding DEGs/DEPs, by which, a better understanding of the mechanisms of submergence tolerance in soybean may be achieved.

Construction of a high-density genetic map and mapping of QTLs for soybean (Glycine max) agronomic and seed quality traits by specific length amplified fragment sequencing.

BACKGROUND: Soybean is not only an important oil crop, but also an important source of edible protein and industrial raw material. Yield-traits and quality-traits are increasingly attracting the attention of breeders. Therefore, fine mapping the QTLs associated with yield-traits and quality-traits of soybean would be helpful for soybean breeders. In the present study, a high-density linkage map was constructed to identify the QTLs for the yield-traits and quality-traits, using specific length amplified fragment sequencing (SLAF-seq). RESULTS: SLAF-seq was performed to screen SLAF markers with 149 F8:11 individuals from a cross between a semi wild soybean, 'Huapidou', and a cultivated soybean, 'Qihuang26', which generated 400.91 M paired-end reads. In total, 53,132 polymorphic SLAF markers were obtained. The genetic linkage map was constructed by 5111 SLAF markers with segregation type of aa×bb. The final map, containing 20 linkage groups (LGs), was 2909.46 cM in length with an average distance of 0.57 cM between adjacent markers. The average coverage for each SLAF marker on the map was 81.26-fold in the male parent, 45.79-fold in the female parent, and 19.84-fold average in each F8:11 individual. According to the high-density map, 35 QTLs for plant height (PH), 100-seeds weight (SW), oil content in seeds (Oil) and protein content in seeds (Protein) were found to be distributed on 17 chromosomes, and 14 novel QTLs were identified for the first time. The physical distance of 11 QTLs was shorter than 100 Kb, suggesting a direct opportunity to find candidate genes. Furthermore, three pairs of epistatic QTLs associated with Protein involving 6 loci on 5 chromosomes were identified. Moreover, 13, 14, 7 and 9 genes, which showed tissue-specific expression patterns, might be associated with PH, SW, Oil and Protein, respectively. CONCLUSIONS: With SLAF-sequencing, some novel QTLs and important QTLs for both yield-related and quality traits were identified based on a new, high-density linkage map. Moreover, 43 genes with tissue-specific expression patterns were regarded as potential genes in further study. Our findings might be beneficial to molecular marker-assisted breeding, and could provide detailed information for accurate QTL localization.

Value of EZSCAN parameters for diabetes screening in Chinese. / Valor de los parámetros de EZSCAN para el cribado de diabetes en la población china.

OBJECTIVE: To study the parameters of EZSCAN as a screening tool for diabetes in Chinese. METHODS: A total of 6,270 subjects participated in the study. All subjects underwent tests of EZSCAN, fasting plasma glucose (FPG), oral glucose tolerance test and HbA1c. RESULTS: 1. All subjects were divided into 4 groups: the normal group, sugar metabolic abnormalities as low-risk group, middle-risk group and high-risk group. The difference of diabetes incidence among the 4 groups was statistically significant. With the increase of EZSCAN score, the prevalence of diabetes increased significantly. But there is no statistically difference between the low-risk group and the middle-risk group. 2. After adjustment for other variables, there is significantly positive relationship among EZSCAN risk score and the risk of diabetes. Meanwhile there is no statistically difference between the low-risk group and the middle-risk group. 3. The cut-off point of EZSCAN for diabetes was 44.5% with the sensitivity was 73.2% which was higher than of FPG and HbA1c. CONCLUSION: As EZSCAN-diabetes risk score increases, the risk of diabetes increases. EZSCAN can be used as a tool for screening for diabetes. At the best screening diabetes cut-off point value 44.5%, the sensitivity is higher than traditional method of FPG and HbA1c.

Effect of pyrolysis conditions on the characteristics of biochar produced from a tobacco stem.

To investigate the potential use of tobacco stems as feedstock for biochar production, a variety of pyrolysis conditions were studied to determine their effects on the distribution of pyrolysis products, as well as the physicochemical properties, energy yield and porosity characteristics of the biochar. The results showed that a moderate temperature of 400-450°C, a retention time of three hours and a slow heating rate of 5°C min(-1) was better to get high energy yield from biochar. In this study, the highest calorific value of biochar was 28.128 MJ kg(-1) The highest biochar yield occurred at a moderate retention time of three hours. The heating rate had an important influence on the characteristics of biochar, and the physiochemical properties of biochar produced at a heating rate of 10°C min(-1) were better than other levels. Also, the oil yield decreased and gas yield increased as retention time and heating rate increased. Biochar produced at a high temperature of 600°C, a retention time up to four hours and a heating rate of 10-15°C min(-1) had a larger BET surface area and cumulative pore volume than that of others. The highest change rate of pore volume of biochar almost occurred at the pore size of 2-5 nm.

RLS3, a protein with AAA+ domain localized in chloroplast, sustains leaf longevity in rice.

Leaf senescence plays an important role in crop developmental processes that dramatically affect crop yield and grain quality. The genetic regulation of leaf senescence is complex, involving many metabolic and signaling pathways. Here, we identified a rapid leaf senescence 3 (rls3) mutant that displayed accelerated leaf senescence, shorter plant height and panicle length, and lower seed set rate than the wild type. Map-based cloning revealed that RLS3 encodes a protein with AAA+ domain, localizing it to chloroplasts. Sequence analysis found that the rls3 gene had a single-nucleotide substitution (G→A) at the splice site of the 10th intron/11th exon, resulting in the cleavage of the first nucleotide in 11th exon and premature termination of RLS3 protein translation. Using transmission electron microscope, the chloroplasts of the rls3 mutant were observed to degrade much faster than those of the wild type. The investigation of the leaf senescence process under dark incubation conditions further revealed that the rls3 mutant displayed rapid leaf senescence. Thus, the RLS3 gene plays key roles in sustaining the normal growth of rice, while loss of function in RLS3 leads to rapid leaf senescence. The identification of RLS3 will be helpful to elucidate the mechanisms involved in leaf senescence in rice.

Receptor-selective IL-4 mutein modulates inflammatory vascular cell phenotypes and attenuates atherogenesis in apolipoprotein E-knockout mice.

The therapeutic potential of interleukin-4-mediated immunomodulation has not been proven in atherogenesis. Type I IL-4 receptor consists of IL-4Rα and a common γ chain, whereas type II IL-4R is a heterodimer of IL-4Rα and IL-13Rα1. Reportedly, the human IL-4 mutein IL-4/R121E is able to act as an IL-4RI-specific agonist. Here, we investigated the effect of receptor-specific IL-4 mutein on vascular cell phenotypes and atherogenesis. Initially, a plasmid expressing murine IL-4/Q116E, analogous to human IL-4/R121E, was transfected to vascular lineage cells in-vitro. IL-4/Q116E induced the activation of STAT6 in b.End3 endothelial cells, Mm1 macrophages, and splenocytes isolated from C57BL6/J (B6) mice, but it failed to activate STAT6 in SMC and J774.1 macrophages. IL-4/Q116E induced the expression of vascular cell adhesion protein-1 in b.End3 cells but not in SMC. IL-4/Q116E did not exhibit pro-inflammatory actions in either macrophage cell line. Splenocytes were also infected with an adenovirus vector expressing IL-4/Q116E (AdIL-4/Q116E). Enzyme-linked immunosorbent assay for interferon-γ, IL-10 and IL-13 revealed that AdIL-4/Q116E-infected splenocytes showed Th2 deviation. Th2 deviation and M2 marker up-regulation were further revealed in ex-vivo assays using the splenocytes isolated from AdIL-4/Q116E-infected apolipoprotein-E knockout (ApoEKO) mice. Finally, adenoviral induction of IL-4/Q116E, but not wild type IL-4, double mutein IL-4/Q116D/Y119D or control ß-galactosidase, significantly attenuated in-vivo atherogenesis of ApoEKO mice. Our data suggest that IL-4 signaling plays a pivotal role in the regulation of vascular cell phenotypes, and atherogenesis. The IL-4RI-selective mutein IL-4/Q116E may have therapeutic potential in vascular diseases.

Vascular smooth muscle cells isolated from adipose triglyceride lipase-deficient mice exhibit distinct phenotype and phenotypic plasticity.

The alteration of triglyceride (TG) metabolism in vascular smooth muscle cells (SMC) is likely to be correlated with certain phenotype, though this has not been elucidated. Adipose triglyceride lipase (ATGL) exerts major TG catalytic activity in both adipotic and non-adipotic cells. In the present study, we isolated SMC from ATGL-deficient mice (ATGL(-/-)mSMC). ATGL(-/-)mSMC showed spontaneous TG accumulation with lower mitogenic response and smooth muscle actin (SMA) expression compared to ATGL (+/+)mSMC. Percentage of senescence-associated ß-galactosidase positive cells was also increased in ATGL(-/-)mSMC. Real-time PCR followed by screening with focused DNA array analysis revealed up-regulated expression of glucokinase (1.7-fold), lipoprotein lipase (3.8-fold) and interleukin-6 (3.7-fold) and down-regulated expression of vascular endothelial growth factor-A (0.2-fold), type I collagen (0.5-fold), and transforming growth factor-ß (0.4-fold) in ATGL(-/-)mSMC compared to ATGL(+/+)mSMC. Next, ectopic gene transfer of human ATGL was attempted using doxycycline (Dox)-regulatable myc-DDK-tagged adenovirus vector (AdvATGL). AdvATGL infection resulted in a reduction of TG accumulation with elevated mitogenic response and SMA expression, and decreased in senescent cell numbers in ATGL(-/-)mSMC. Moreover, deviated gene expression pattern in ATGL(-/-)mSMC was potentially corrected. Our data suggest that ATGL(-/-)mSMC have a distinct phenotype that may be related to vascular pathogenesis. Plasticity of SMC phenotypes correlated to lipid metabolism could be a therapeutic target.