Explore the mechanism of incomplete Kawasaki disease and identify a novel biomarker by weighted gene co-expression network analysis.

Incomplete Kawasaki Disease is a complex disease that often occurs in infants and has substantial coronary artery damage. Its pathogenesis is unclear and lacks specific diagnostic markers. The purpose of our study is to research the mechanism of incomplete Kawasaki Disease use of bioinformatic methods and identify potential biomarkers. We performed weighted gene co-expression network analysis to analyze the data set GSE68004 and identified modules and genes which were correlated with the disease. Through functional annotation and enrichment analysis, we determined the biological function and signal pathway of these genes. We further used lasso regression and ROC curve to screen genes and determined that the final candidate gene was HSPB11and hsa-miR-155-5p that regulates its expression. Finally, we validated the screened gene using an independent dataset and construct a TF-miRNA network. Through the relationships of TFs and hsa-miR-155-5p, we found is hsa-miR-155-5p closely related to hypoxia-related transcription factors, which may be a new direction in the research of Kawasaki disease.

An epigenetic basis of inbreeding depression in maize.

Inbreeding depression is widespread across plant and animal kingdoms and may arise from the exposure of deleterious alleles and/or loss of overdominant alleles resulting from increased homozygosity, but these genetic models cannot fully explain the phenomenon. Here, we report epigenetic links to inbreeding depression in maize. Teosinte branched1/cycloidea/proliferating cell factor (TCP) transcription factors control plant development. During successive inbreeding among inbred lines, thousands of genomic regions across TCP-binding sites (TBS) are hypermethylated through the H3K9me2-mediated pathway. These hypermethylated regions are accompanied by decreased chromatin accessibility, increased levels of the repressive histone marks H3K27me2 and H3K27me3, and reduced binding affinity of maize TCP-proteins to TBS. Consequently, hundreds of TCP-target genes involved in mitochondrion, chloroplast, and ribosome functions are down-regulated, leading to reduced growth vigor. Conversely, random mating can reverse corresponding hypermethylation sites and TCP-target gene expression, restoring growth vigor. These results support a unique role of reversible epigenetic modifications in inbreeding depression.

Acute appendicitis in a child with nonspecific signs and symptoms and nondiagnostic sonography: Necessity of computed tomography.

Acute appendicitis is a common acute abdomen in children, especially in children over 5 years old. Although the incidence rate is lower than that of adults, the disease is more serious than adults. The rate of complication of peritonitis and perforation of appendix is even high. Generally, abdominal pain is still the main symptom of acute appendicitis in children, but children cannot express it, parents and doctors are easy to neglect. Ultrasound is the most commonly used imaging examination in the diagnosis of acute appendicitis in children. High fever can appear earlier, up to 39°C, at the same time can have mental atrophy, chills, convulsions and toxic shock. However, when the clinical symptoms of patients are not typical, and no obvious abnormality is found by ultrasound, it is easy to cause misdiagnosis to clinicians. Here we report a case of 9-years-old Chinese female with intermittent abdominal pain and vomiting. Initially she was diagnosed with acute gastroenteritis and was treated with antibiotics. However, there was nothing found by ultrasound, and her abdominal pain symptoms still did not relieve. Finally, abdominal CT examination confirmed acute appendicitis after 48 hours. The lesson is that ultrasound scanning should not be limited to the right lower abdomen, due to the great variation of appendix position in children with appendicitis. In addition, if there is no abnormality found by ultrasound and the infection index is increased, we suggest that abdominal CT should be examined immediately.

Heterologous expression of ZmGS5 enhances organ size and seed weight by regulating cell expansion in Arabidopsis thaliana.

Maize ZmGS5 was reported to be positively associated with kernel-related traits, however, its regulatory mechanism on plant development and seed size remains unknown. In this study, ZmGS5 was demonstrated to be widely expressed in various maize tissues with the highest expression level in developing embryos, indicating its critical roles in early kernel development process. The ZmGS5 protein was subcellularly localized to both the nucleus and cytoplasm. Transgenic Arabidopsis plants overexpressing ZmGS5 under the control of either the constitutive maize Ubiquitin1 promotor or native ZmGS5 promoter resulted in increased plant size, biomass, seed size and weight, although no significant difference was observed between transgenic lines harboring the two constructs. In contrast, the antisense-ZmGS5 transgene resulted in opposite phenotypes. Our cytological data suggested that ZmGS5 enlarged petal size through enhancing cell expansion. Quantitative RT-PCR analysis indicated that ZmGS5 might enhance cell expansion and grain filling by upregulating expression levels of particular EXPA or SWEET genes. Collectively, these findings help us further understand the biological function and regulatory mechanism of ZmGS5 in improving organ size and seed weight, which imply its great potential for high-yield breeding in the future.

Overexpression of an Antisense RNA of Maize Receptor-Like Kinase Gene ZmRLK7 Enlarges the Organ and Seed Size of Transgenic Arabidopsis Plants.

Leucine-rich repeat (LRR)-receptor-like protein kinases (LRR-RLKs) play vital roles in plant growth, development, and responses to environmental stresses. In this study, a new LRR-RLK gene, ZmRLK7, was isolated from maize, and its function within plant development was investigated through ectopic expression in Arabidopsis. The spatial expression pattern analysis reveals that ZmRLK7 is highly expressed in embryos prior to programmed cell death (PCD) of starchy endosperm tissues, and its encoded protein has been localized to both plasm and nuclear membranes subcellularly. Overexpression of sense ZmRLK7 reduced the plant height, organ size (e.g., petals, silique, and seeds), and 1000-seed weight in transgenic lines, while the antisense transgene enlarged these traits. Cytological analysis suggested that ZmRLK7 negatively regulates petal size through restricting both cell expansion and proliferation. In addition, abnormal epidermal cell structure was observed, and the stomata number decreased obviously in sense ZmRLK7 transgenic lines with a lower stomatal index than that in the wild type. Quantitative RT-PCR analysis indicated that transcript levels of genes that are involved in the brassinosteroid and ERACTA signaling pathways were coordinately altered, which could partially explain the phenotypic variation. Moreover, overexpression of antisense ZmRLK7 substantially rescued the Arabidopsis bak1-3 mutant phenotype. All these results together suggest that ZmRLK7 can serve as an important regulator in regulating plant architecture and organ size formation. This work will provide insight into the function of ZmRLK7 in maize.

Overexpression of maize sucrose non-fermenting-1-related protein kinase 1 genes, ZmSnRK1s, causes alteration in carbon metabolism and leaf senescence in Arabidopsis thaliana.

Sucrose non-fermenting-1 (SNF1) -related protein kinase 1 (SnRK1) is a key regulator of catabolic homeostasis and plays critical roles in plant development and stress response. In this study, three SNF1-related protein kinase 1 genes, ZmSnRK1.1, ZmSnRK1.2 and ZmSnRK1.3, which are highly conserved in plants, were isolated from maize (Zea mays L.). Expression profiling experiments indicated that the three genes were constitutively expressed in all tested tissues with the highest expression level in young ears. Subcellular localization analysis indicated that ZmSnRK1.1, ZmSnRK1.2 and ZmSnRK1.3 are localized to both the nucleus and cytoplasm. Transgenic Arabidopsis lines overexpressing ZmSnRK1.1, ZmSnRK1.2 or ZmSnRK1.3 exhibited hypersensitivity to exogenous sugar treatment and accumulated less glucose but more sucrose in the rosette leaves and mature seeds compared to the wild type. Time to flowering was shortened in the ZmSnRK1.1 over-expressing lines but prolonged in the ZmSnRK1.2 and ZmSnRK1.3 lines. Leaf senescence was delayed in all transgenic lines, especially in the ZmSnRK1.3 lines, which led to enhanced biomass and seed yield at maturity. Key genes that are involved in carbon metabolism, senescence and flowering time were differentially regulated in the transgenic lines as revealed by the RNA-seq analysis. This study demonstrated that maize ZmSnRK1 members play important roles in energy sensing and carbon metabolism, they regulate the architecture shaping and developmental transition when heterogeneously expressed in Arabidopsis and may provide potentially valuable characteristics for high yield breeding of crops in the future.

Fine Mapping and Candidate Gene Analysis of the Leaf-Color Gene ygl-1 in Maize.

A novel yellow-green leaf mutant yellow-green leaf-1 (ygl-1) was isolated in self-pollinated progenies from the cross of maize inbred lines Ye478 and Yuanwu02. The mutant spontaneously showed yellow-green character throughout the lifespan. Meanwhile, the mutant reduced contents of chlorophyll and Car, arrested chloroplast development and lowered the capacity of photosynthesis compared with the wild-type Lx7226. Genetic analysis revealed that the mutant phenotype was controlled by a recessive nuclear gene. The ygl-1 locus was initially mapped to an interval of about 0.86 Mb in bin 1.01 on the short arm of chromosome 1 using 231 yellow-green leaf individuals of an F2 segregating population from ygl-1/Lx7226. Utilizing four new polymorphic SSR markers, the ygl-1 locus was narrowed down to a region of about 48 kb using 2930 and 2247 individuals of F2 and F3 mapping populations, respectively. Among the three predicted genes annotated within this 48 kb region, GRMZM2G007441, which was predicted to encode a cpSRP43 protein, had a 1-bp nucleotide deletion in the coding region of ygl-1 resulting in a frame shift mutation. Semi-quantitative RT-PCR analysis revealed that YGL-1 was constitutively expressed in all tested tissues and its expression level was not significantly affected in the ygl-1 mutant from early to mature stages, while light intensity regulated its expression both in the ygl-1 mutant and wild type seedlings. Furthermore, the mRNA levels of some genes involved in chloroplast development were affected in the six-week old ygl-1 plants. These findings suggested that YGL-1 plays an important role in chloroplast development of maize.

Anchoring dental implant in tissue-engineered bone using composite scaffold: a preliminary study in nude mouse model.

PURPOSE: The purpose of this study was to fabricate a tissue-engineered bone graft anchoring dental implant with bone marrow stromal cell (bMSC) seeded coral-implant composite scaffold. MATERIALS AND METHODS: Titanium dental implants (3 mm in diameter) were inserted into the cylinder coral scaffolds (5 mm in diameter and 1 mm in wall thickness). bMSCs were isolated from iliac bone marrow of adult New Zealand White rabbits, induced by dexamethasone and seeded into the composite scaffold at the density of 2 x 10 8 /mL in 200 muL medium. Nine cell coral-implant complexes were incubated in vitro for 5 days. One complex was processed for scanning electronic microscopy. The other 8 complexes, together with 4 coral scaffold without cell acting as control, were implanted subcutaneously into nude mice back. At 1 and 2 months after implantation, 4 specimens from the experiment group and 2 specimens from the control group were harvested respectively. New bone restoration and new bone integration with dental implant were evaluated by gross inspection, manual handling test, radiographic examination, and histologic observation. RESULTS: Specimens harvested at 2 months after implantation were red and similar to native bone. Manual handling test showed that dental implants were fixed in the newly formed bone. Radiographic examination showed that most of the coral scaffold had been absorbed. Bone density x-ray shadow could be observed around the dental implant. Histologic examination showed that large amount of new bone formed around the dental implants and integrated well with the implants in some area. In the control group no bone formation was observed both macroscopically and microscopically. CONCLUSION: The results of the study suggested that the tissue-engineered bone of bMSCs seeded natural coral-implant composite scaffold is promising for dental implant anchoring, which has positive implication for clinical jaw reconstruction.