Mdm-miR160-MdARF17-MdWRKY33 module mediates freezing tolerance in apple.

Apple (Malus domestica) trees are vulnerable to freezing temperatures. Cold resistance in woody perennial plants can be improved through biotechnological approaches. However, genetic engineering requires a thorough understanding of the molecular mechanisms of the tree's response to cold. In this study, we demonstrated that the Mdm-miR160-MdARF17-MdWRKY33 module is crucial for apple freezing tolerance. Mdm-miR160 plays a negative role in apple freezing tolerance, whereas MdARF17, one of the targets of Mdm-miR160, is a positive regulator of apple freezing tolerance. RNA sequencing analysis revealed that in apple, MdARF17 mediates the cold response by influencing the expression of cold-responsive genes. EMSA and ChIP-qPCR assays demonstrated that MdARF17 can bind to the promoter of MdWRKY33 and promotes its expression. Overexpression of MdWRKY33 enhanced the cold tolerance of the apple calli. In addition, we found that the Mdm-miR160-MdARF17-MdWRKY33 module regulates cold tolerance in apple by regulating reactive oxygen species (ROS) scavenging, as revealed by (i) increased H2 O2 levels and decreased peroxidase (POD) and catalase (CAT) activities in Mdm-miR160e OE plants and MdARF17 RNAi plants and (ii) decreased H2 O2 levels and increased POD and CAT activities in MdmARF17 OE plants and MdWRKY33 OE calli. Taken together, our study uncovered the molecular roles of the Mdm-miR160-MdARF17-MdWRKY33 module in freezing tolerance in apple, thus providing support for breeding of cold-tolerant apple cultivars.

The RNA-binding protein MdHYL1 modulates cold tolerance and disease resistance in apple.

Apple (Malus domestica) trees often experience various abiotic and biotic stresses. However, due to the long juvenile period of apple and its high degree of genetic heterozygosity, only limited progress has been made in developing cold-hardy and disease-resistant cultivars through traditional approaches. Numerous studies reveal that biotechnology is a feasible approach to improve stress tolerance in woody perennial plants. HYPONASTIC LEAVES1 (HYL1), a double-stranded RNA-binding protein, is a key regulator involved in apple drought stress response. However, whether HYL1 participates in apple cold response and pathogen resistance remains unknown. In this study, we revealed that MdHYL1 plays a positive role in cold tolerance and pathogen resistance in apple. MdHYL1 acted upstream to positively regulate freezing tolerance and Alternaria alternata resistance by positively modulating transcripts of MdMYB88 and MdMYB124 in response to cold stress or A. alternata infection. In addition, MdHYL1 regulated the biogenesis of several miRNAs responsive to cold and A. alternata infection in apple. Furthermore, we identified Mdm-miRNA156 (Mdm-miR156) as a negative regulator of cold tolerance and Mdm-miRNA172 (Mdm-miR172) as a positive regulator of cold tolerance, and that Mdm-miRNA160 (Mdm-miR160) decreased plant resistance to infection by A. alternata. In summary, we highlight the molecular role of MdHYL1 regarding cold tolerance and A. alternata infection resistance, thereby providing candidate genes for breeding apple with freezing tolerance and A. alternata resistance using biotechnology.

First Report of Colletotrichum fioriniae Causing Anthracnose on Persimmon in China.

Persimmons (Diospyros kaki Thunb.) have a longstanding history of cultivation in China. Both aesthetically pleasing and edible, they often symbolize a sweet and fulfilling life. During the summer of 2022, a severe outbreak of anthracnose was observed on the lower leaves of persimmon trees in the National Field Genebank for Persimmon (NFGP), located in Yangling, Shaanxi, China (34°17'42.80″ N, 108°04'08.21″ E). The estimated incidence rate of this disease within the NFGP was approximately 30%. The typical symptoms of the disease included the presence of irregular lesions on leaves, and oval sunken lesions on infected fruit. Under high humidity conditions, pink sticky substances appeared in the affected areas. The presence of numerous lesions led to softening and detachment of persimmon fruit. To identify the causal pathogen, 5 × 5mm samples of the diseased leaves were collected from the interface between the infected and healthy leaves. The leaves were disinfected with 70% alcohol for 20 s, followed by rinsing with sterile water. Subsequently, the leaves were immersed in 1% NaClO for 2 to 3 minutes, rinsed with sterile water three times, dried using sterile absorbent paper, and the leaf samples were then transferred onto potato dextrose agar (PDA) medium, and cultured in 25°C incubators. Once the colony reached a certain size, small pieces of hyphae were extracted from edge and transferred for purification and repeated three times. After being cultured on PDA for 7 days, the colony showed a white spongy surface with a pink-orange center. The conidia displayed a fusiform shape and were transparent, measuring 4.58 to 6.53 µm × 9.27 to 13.11 µm (n=50). The isolates share morphological similarities with Colletotrichum fioriniae. The representative isolate HY-7 was selected for molecular identification. The internal transcribed spacers (ITS) region, chitin synthase (CHS-1), actin (ACT), beta-tubulin 2 (TUB2), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene were amplified using ITS1/4 (White et al. 1990), CHS-79F/CHS-345R (Carbone & Kohn, 1999), ACT512F/ACT (Carbone & Kohn, 1999), T1/BT2B (Glass & Donaldson 1995, O'Donnell et al., 1997), and GDF/GDR (Templeton et al. 1992), respectively. The generated sequences were deposited at GenBank under accession numbers OR878056 (ITS), OR766019 (CHS-1), OR766021(TUB2), OR766018 (ACT) and OR766020 (GAPDH). BLAST analysis revealed the sequences were 100% identical to C. fioriniae (MH865005 for ITS, JQ948953 for CHS-1, JQ949613 for ACT, JQ949943 for TUB2 and JQ948622 for GAPDH). The morphological characteristics and molecular analyses of the isolate matched the description of C. fioriniae. To fulfill Koch's postulates, the twigs and leaves of 'Fupingjianshi' in four different directions were inoculated without wounding in the field, and 10 healthy fruits were selected for wound inoculation. The concentration of conidia used for inoculation was about 1 × 106 conidia/ml, and sterilized water was used as control. The experiment was replicated three times under the same conditions. One week after inoculation, characteristic symptoms resembling those observed on the leaves of primary diseased persimmon trees appeared on the leaves and fruits. No symptoms were observed on the leaves, twigs and fruits in the control treatment. The pathogen from the artificially infected leaves and fruits were reisolated and identified as C. fiorinae based on morphological and molecular characteristics. Persimmon anthracnose is a common disease in regions where the fruit is grown, to the best of our knowledge, this is the first documented occurrence of C. fioriniae-induced anthracnose on persimmons in China, which should be paid more attentions. This report will help identify disease symptoms in the field and provides a basis for determining the occurrence, distribution, and control of C. fioriniae on persimmon leaves and fruits.

Integration of land ecological consolidation and ecosystem product value realization: A case from the Yangtze riverside industrial park in Changzhou, China.

Traditional industries and industrialization have led to widespread environmental pollution and ecosystem degradation in major river basins globally. Strategies centered on ecological restoration and ecological economy are emerging as essential tools for effective environmental governance. This study aims to investigate how a multifaceted framework for land ecological consolidation, with various developmental goals, can effectively support ecological restoration and sustainability. Through quantitative analysis and in-depth interviews, we investigated the case of Yangtze riverside chemical industrial park in Changzhou. This park pursues ecological and economic sustainability through chemical industry transformation, ecological restoration and protection, ecological management, and ecological industry development. The results show that this practice established a multi-objective action framework rooted in urban renewal, land consolidation, ecological restoration, industrial transformation, and rural revitalization. Through multiplanning integration, integrated implementation and full-cycle profit distribution, the aim of ecological protection has been initially achieved, offering a crucial guarantee for sustainable development. A total of 96.47 ha ecological space expanded, which can generate ecological product worth CNY 7.283 billion, alongside a net economic benefit of CNY 978 million over three decades. The top-down ecological responsibilities, coupled with local developmental demands, have stimulated collaborations within a bottom-up endogenous network comprising government, enterprises, and residents.

MdGH3.6 is targeted by MdMYB94 and plays a negative role in apple water-deficit stress tolerance.

Drought significantly limits apple fruit production and quality. Decoding the key genes involved in drought stress tolerance is important for breeding varieties with improved drought resistance. Here, we identified GRETCHEN HAGEN3.6 (GH3.6), an indole-3-acetic acid (IAA) conjugating enzyme, to be a negative regulator of water-deficit stress tolerance in apple. Overexpressing MdGH3.6 reduced IAA content, adventitious root number, root length and water-deficit stress tolerance, whereas knocking down MdGH3.6 and its close paralogs increased IAA content, adventitious root number, root length and water-deficit stress tolerance. Moreover, MdGH3.6 negatively regulated the expression of wax biosynthetic genes under water-deficit stress and thus negatively regulated cuticular wax content. Additionally, MdGH3.6 negatively regulated reactive oxygen species scavengers, including antioxidant enzymes and metabolites involved in the phenylpropanoid and flavonoid pathway in response to water-deficit stress. Further study revealed that the homolog of transcription factor AtMYB94, rather than AtMYB96, could bind to the MdGH3.6 promoter and negatively regulated its expression under water-deficit stress conditions in apple. Overall, our results identify a candidate gene for the improvement of drought resistance in fruit trees.

The positive feedback regulatory loop of miR160-Auxin Response Factor 17-HYPONASTIC LEAVES 1 mediates drought tolerance in apple trees.

Drought stress tolerance is a complex trait regulated by multiple factors. Here, we demonstrate that the miRNA160-Auxin Response Factor 17 (ARF17)-HYPONASTIC LEAVES 1 module is crucial for apple (Malus domestica) drought tolerance. Using stable transgenic plants, we found that drought tolerance was improved by higher levels of Mdm-miR160 or MdHYL1 and by decreased levels of MdARF17, whereas reductions in MdHYL1 or increases in MdARF17 led to greater drought sensitivity. Further study revealed that modulation of drought tolerance was achieved through regulation of drought-responsive miRNA levels by MdARF17 and MdHYL1; MdARF17 interacted with MdHYL1 and bound to the promoter of MdHYL1. Genetic analysis further suggested that MdHYL1 is a direct downstream target of MdARF17. Importantly, MdARF17 and MdHYL1 regulated the abundance of Mdm-miR160. In addition, the Mdm-miR160-MdARF17-MdHYL1 module regulated adventitious root development. We also found that Mdm-miR160 can move from the scion to the rootstock in apple and tomato (Solanum lycopersicum), thereby improving root development and drought tolerance of the rootstock. Our study revealed the mechanisms by which the positive feedback loop of Mdm-miR160-MdARF17-MdHYL1 influences apple drought tolerance.

Microstructural Alterations in Projection and Association Fibers in Neonatal Hypoxia-Ischemia.

BACKGROUND: Diffusion MRI (dMRI) is known to be sensitive to hypoxic-ischemic encephalopathy (HIE). However, existing dMRI studies used simple diffusion tensor metrics and focused only on a few selected cerebral regions, which cannot provide a comprehensive picture of microstructural injury. PURPOSE: To systematically characterize the microstructural alterations in mild, moderate, and severe HIE neonates compared to healthy neonates with advanced dMRI using region of interest (ROI), tract, and fixel-based analyses. STUDY TYPE: Prospective. POPULATION: A total of 42 neonates (24 males and 18 females). FIELD STRENGTH/SEQUENCE: 3-T, diffusion-weighted echo-planar imaging. ASSESSMENT: Fractional anisotropy (FA), mean diffusivity (MD), radial diffusivity (RD), axial diffusivity (AD), fiber density (FD), fiber cross-section (FC), and fiber density and cross-section (FDC) were calculated in 40 ROIs and 6 tracts. Fixel-based analysis was performed to assess group differences in individual fiber components within a voxel (fixel). STATISTICAL TESTS: One-way analysis of covariance (ANCOVA) to compare dMRI metrics among severe/moderate/mild HIE and control groups and general linear model for fixel-wise group differences (age, sex, and body weight as covariates). Adjusted P value < 0.05 was considered statistically significant. RESULTS: For severe HIE, ROI-based analysis revealed widespread regions, including the deep nuclei and white matter with reduced FA, while in moderate injury, only FC was decreased around the posterior watershed zones. Tract-based analysis demonstrated significantly reduced FA, FD, and FC in the right inferior fronto-occipital fasciculus (IFOF), right inferior longitudinal fasciculus (ILF), and splenium of corpus callosum (SCC) in moderate HIE, and in right IFOF and left anterior thalamic radiation (ATR) in mild HIE. Correspondingly, we found altered fixels in the right middle-posterior IFOF and ILF, and in the central-to-right part of SCC in moderate HIE. DATA CONCLUSION: For severe HIE, extensive microstructural injury was identified. For moderate-mild HIE, association fiber injury in posterior watershed area with a rightward lateralization was found. EVIDENCE LEVEL: 1 TECHNICAL EFFICACY: Stage 3.

First Report of Leaf Black Spot Caused by Alternaria alternata on American Persimmons in China.

American persimmons (Diospyros virginiana L.) are native to the United States. After being introduced into China, they were used as a rootstock for expanding persimmon varieties and planted in local areas due to their strong cold resistance and diverse leaf colors. In 2022, 12 plants had similar symptoms to black spot disease on the leaves of 18 American persimmons introduced in the National Field Genebank for Persimmon, Yangling, Shaanxi, China (34°17'42.80″ N, 108°04'08.21″ E). Among them, severity was highest in the 'VM10' variety (almost 100%), 'VM10' is the main cultivar in Shaanxi and Henan regions of China, and the incidence of disease in the two regions ranged between 30 and 60% in 2022. Early symptoms were irregular black-brown spots, which gradually combined into large irregular lesions with a dark brown border. The leaves began to curl, crack, scorch, and abscissed. When relative humidity was high, leaves also had signs of black sporulation and became chlorotic. To isolate the causal agent, 10 symptomatic leaves were collected from 5 diseased plants in the National Field Genebank for Persimmon. The infected leaves were cut into 20 small pieces of 5 × 5 mm from the junction of the diseased and healthy tissues and surface disinfected in 75% alcohol for 15 sec, washed with sterile water and 2% NaClO for 90 sec, rinsed three times with sterile water, dried with sterile absorbent paper, and plated on potato dextrose agar (PDA) medium. After 3 days, 12 strains of fungi were isolated from the tissue by transferring the hyphal tips of the mycelium. Among them, 10 strains had similar morphological characteristics. Fungal colonies developed on the PDA medium were initially white, then gradually changed to gray-brown with neat edges and flocculent hyphae. Conidia (n=50) light brown or medium brown, obovate or pear-shaped, and 8.27 to 15.31×17.51 to 24.31 µm, with 1 to 4 transverse septa and 0 to 2 longitudinal septa. The isolates were morphologically similar to Alternaria alternata (Simmons et al. 2007). For molecular identification, the E.Z.N.A.® Fungal DNA kit (Omega Bio-Tek) was used to extract genomic DNA from 7-day-old mycelium grown on PDA medium. The internal transcribed spacers (ITS) region, translation elongation factor 1-alpha (TEF1-α), Alternaria major allergen (Alt a1) gene, and partial RNA polymerase second largest subunit (RPB2) were amplified using ITS1/4 (Glass et al. 1995), EF1-728F/EF1-968R (Carbone and Kohn 1999), and Alt-4for /Alt-4rev (Hong et al., 2005) and RPB2-5F/RPB2-7CR (Liu et al. 1999) respectively. The sequences of a representative isolate MZS1 were deposited in GeneBank with accession numbers OP198643 for ITS, OP286949 for Tef1-α, OP286948 for Alt a1, and OP951084 for RPB2, and were 100% identical to strains of A. alternata (MN615420 for ITS, MN615423 for EF1-α, MW848792 for Alt a1 and MN615422 for RPB2). A maximum-likelihood (ML) phylogenetic tree was constructed based on the concatenated sequences of ITS, TEF1-α, Alt a1, and RPB2gene, which clustered with the A. alternata strains YZU191238 with high bootstrap support (99%). To fulfill Koch's postulates, three-month-old American persimmon 'VM10' seedlings were tested for pathogenicity. Three seedlings were sprayed with 1 × 106 spores/ml suspension in a spray pot, and the three seedlings were treated with sterilized water as a noninoculated control. All seedlings were cultured in a 25°C incubator. The experiment was performed three times under the same conditions. One week after inoculation, typical symptoms appeared on the leaves, which were similar to those observed on the leaves of the original infected persimmon trees. In the control treatment, the leaves did not show symptoms. To the best of our knowledge, this is the first report of A. alternata causing American persimmon black spots disease in China. This report will contribute to the identification of disease symptoms in the field and provide a basis for the occurrence, distribution, and control of A. alternata on American persimmon leaves.

Cancer and its predictors in Chinese adults with newly diagnosed diabetes and impaired glucose tolerance (IGT): a 30-year follow-up of the Da Qing IGT and Diabetes Study.

BACKGROUND: We aimed to explore if hyperglycaemia and hyperinsulinemia in the diabetes and prediabetes population were associated with increased risk of cancer occurence. METHODS: Overall, 1700 participants with different glycaemic statuses were screened from the 110,660 residents of Da-Qing, China, in 1985. They were followed up to 30 years to access cancer outcomes. RESULTS: Cancer was identified in 15.2% (259/1700) of the participants. The incidence of cancer in the normal glucose tolerance (NGT), impaired glucose tolerance (IGT) and diabetes groups was 6.06, 6.77, and 7.18 per 1000 person-years, respectively (P = 0.02). In the Fine-Gray model with all cause death as competing risk, compared with the NGT controls, both IGT and diabetes groups demonstrated significantly higher risk of cancer (for the IGT group, adjusted hazard ratio (aHR) = 1.77, 95% CI 1.38-2.27, P < 0.0001; for the diabetes, aHR = 3.34, 95% CI 2.64-4.22, P < 0.0001). Among the IGT participants, progress to diabetes (aHR = 2.28, 95%CI 1.24-4.20, P = 0.008) and insulin-area under the curve at baseline (for 1 SD increase, aHR = 1.39, P = 0.02) were also associated with the risk of cancer after adjustment of covariables. CONCLUSIONS: Hyperglycaemia in patients with diabetes, hyperinsulinemia, and progression to diabetes in people with IGT is significantly associated with the long-term increased risk of cancer occurrence.

MdZAT5 regulates drought tolerance via mediating accumulation of drought-responsive miRNAs and mRNAs in apple.

Drought limits apple yield and fruit quality. However, the molecular mechanism of apple in response to drought is not well known. Here, we report a Cys2/His2 (C2H2)-type zinc-finger protein, MdZAT5, that positively regulates apple drought tolerance by regulating drought-responsive RNAs and microRNAs (miRNAs). DNA affinity purification and sequencing and yeast-one hybrid analysis identified the binding motifs of MdZAT5, T/ACACT/AC/A/G. Chromatin immunoprecipitation quantitative polymerase chain reaction (ChIP-qPCR) and electrophoretic mobility shift assays (EMSAs) showed that MdZAT5 directly binds to the promoters of the drought-responsive genes including MdRHA2a, MdLEA14, MdTPX1, and MdCAT3, and activates their expression under drought stress. MdZAT5 interacts with and directly targets HYPONASTIC LEAVES1 (MdHYL1). MdZAT5 may facilitate the interaction of MdHYL1 with pri-miRNAs or MdDCL1 by activating MdHYL1 expression, thereby regulating the biogenesis of drought-responsive miRNAs. Genetic dissection showed that MdHYL1 is essential for MdZAT5-mediated drought tolerance and miRNA biogenesis. In addition, ChIP-qPCR and EMSA revealed that MdZAT5 binds directly to the promoters of some MIR genes including Mdm-miR171i and Mdm-miR172c, and modulates their transcription. Taken together, our findings improve our understanding of the molecular mechanisms of drought response in apple and provide a candidate gene for the breeding of drought-tolerant cultivars.

A genome-wide association study provides insights into fatty acid synthesis and metabolism in Malus fruits.

As a precursor of aromatic compounds, fatty acids play important roles in apple fruit quality; however, the genetic and molecular basis underlying fatty acid synthesis and metabolism is largely unknown. In this study, we conducted a genome-wide association study (GWAS) of seven fatty acids using genomic data of 149 Malus accessions and identified 232 significant signals (-log10P>5) associated with 99 genes from GWAS of four fatty acids across 2 years. Among these, a significant GWAS signal associated with linoleic acid was identified in the transcriptional regulator SUPERMAN-like (SUP) MD13G1209600 at chromosome 13 of M. × domestica. Transient overexpression of MdSUP increased the contents of linoleic and linolenic acids and of three aromatic components in the fruit. Our study provides genetic and molecular information for improving the flavor and nutritional value of apple.

Detecting acute bilirubin encephalopathy in neonates based on multimodal MRI with deep learning.

BACKGROUND: Differentiating acute bilirubin encephalopathy (ABE) from non-ABE in neonates with hyperbilirubinemia (HB) from routine magnetic resonance imaging (MRI) is extremely challenging since both conditions demonstrate similar T1 hyperintensities. To this end, we investigated whether the integration of multimodal MRI from routine clinical scans with deep-learning approaches could improve diagnostic performance. METHODS: A total of 75 neonates with ABE and 75 neonates with HB (non-ABE) were included in the study. Each patient had three types of multimodal images taken, i.e., a T1-weighted image (T1WI), a T2-weighted image (T2WI), and an apparent diffusion coefficient (ADC) map. The three types of MRI contrasts and their combination were fed into two deep convolutional neural networks (CNNs), i.e., ResNet18 and DenseNet201. The performance of CNNs was compared with a traditional statistical method named logistic regression. RESULTS: We demonstrated that diagnostic methods with the multimodal data were better than any of the single-modal data. Both CNN models outperformed the logistic regression method. The best performance was achieved by DenseNet201 with the combination of three modalities of T1WI, T2WI, and ADC, with an accuracy of 0.929 ± 0.042 and an area under the curve (AUC) of 0.991 ± 0.007. CONCLUSIONS: Our study demonstrated that CNN models with multimodal MRI significantly improve the accuracy of diagnosing ABE. IMPACT: We proposed an efficient strategy of detecting ABE in neonates based on multimodal MRI with deep learning, which achieved an accuracy of 0.929 ± 0.042 and an AUC of 0.991 ± 0.007. We demonstrated the advantage of integrating multimodal MRI in detecting ABE in neonates with HB, using deep-learning models. Our strategy of diagnosing ABE using deep-learning techniques with multimodal MRI from routine clinical scans is potentially applicable to clinical practice.

Refractory thrombocytopenia could be a rare initial presentation of Noonan syndrome in newborn infants: a case report and literature review.

BACKGROUND: Noonan syndrome (NS) is a relatively rare inherited disease. Typical clinical presentation is important for the diagnosis of NS. But the initial presentation of NS could be significant variant individually which results in the difficult of working diagnosis. Here we report a rare neonatal case of NS who presented with refractory thrombocytopenia as the initial manifestation. CASE PRESENTATION: This was a preterm infant with refractory thrombocytopenia of unknown origin transferred from obstetric hospital at 6 weeks of age. During hospitalization, typical phenotypes of NS in addition to thrombocytopenia were observed, such as typical facial characteristics, short stature, atrial septal defect, cryptochidism, coagulation defect and chylothorax. Genetic testing showed a pathogenic variant at exon 2 of the PTPN11 gene with c.124A > G (p.T42A). Respiratory distress was deteriorated with progressive chylothorax. Chest tube was inserted for continuous draining. Chemical pleurodesis with erythromycin was tried twice, but barely effective. Finally, parents decided to withdraw medical care and the patient died. CONCLUSIONS: Thrombocytopenia could be the first symptom of Noonan syndrome. After ruling out other common causes of thrombocytopenia, NS should be considered as the working diagnosis.

Insights into the effect of human civilization on Malus evolution and domestication.

The evolutionary history of the Malus genus has not been well studied. In the current study, we presented genetic evidence on the origin of the Malus genus based on genome sequencing of 297 Malus accessions, revealing the genetic relationship between wild species and cultivated apples. Our results demonstrated that North American and East Asian wild species are closer to the outgroup (pear) than Central Asian species, and hybrid species including natural (separated before the Pleistocene, about 2.5 Mya) and artificial hybrids (including ornamental trees and rootstocks) are between East and Central Asian wild species. Introgressions from M. sylvestris in cultivated apples appeared to be more extensive than those from M. sieversii, whose genetic background flowed westward across Eurasia and eastward to wild species including M. prunifolia, M. × asiatica, M. × micromalus, and M. × robust. Our results suggested that the loss of ancestral gene flow from M. sieversii in cultivated apples accompanied the movement of European traders around the world since the Age of Discovery. Natural SNP variations showed that cultivated apples had higher nucleotide diversity than wild species and more unique SNPs than other apple groups. An apple ERECTA-like gene that underwent selection during domestication on 15th chromosome was identified as a likely major determinant of fruit length and diameter, and an NB-ARC domain-containing gene was found to strongly affect anthocyanin accumulation using a genome-wide association approach. Our results provide new insights into the origin and domestication of apples and will be useful in new breeding programmes and efforts to increase fruit crop productivity.

Detecting neonatal acute bilirubin encephalopathy based on T1-weighted MRI images and learning-based approaches.

BACKGROUND: Neonatal hyperbilirubinemia is a common clinical condition that requires medical attention in newborns, which may develop into acute bilirubin encephalopathy with a significant risk of long-term neurological deficits. The current clinical challenge lies in the separation of acute bilirubin encephalopathy and non-acute bilirubin encephalopathy neonates both with hyperbilirubinemia condition since both of them demonstrated similar T1 hyperintensity and lead to difficulties in clinical diagnosis based on the conventional radiological reading. This study aims to investigate the utility of T1-weighted MRI images for differentiating acute bilirubin encephalopathy and non-acute bilirubin encephalopathy neonates with hyperbilirubinemia. METHODS: 3 diagnostic approaches, including a visual inspection, a semi-quantitative method based on normalized the T1-weighted intensities of the globus pallidus and subthalamic nuclei, and a deep learning method with ResNet18 framework were applied to classify 47 acute bilirubin encephalopathy neonates and 32 non-acute bilirubin encephalopathy neonates with hyperbilirubinemia based on T1-weighted images. Chi-squared test and t-test were used to test the significant difference of clinical features between the 2 groups. RESULTS: The visual inspection got a poor diagnostic accuracy of 53.58 ± 5.71% indicating the difficulty of the challenge in real clinical practice. However, the semi-quantitative approach and ResNet18 achieved a classification accuracy of 62.11 ± 8.03% and 72.15%, respectively, which outperformed visual inspection significantly. CONCLUSION: Our study indicates that it is not sufficient to only use T1-weighted MRI images to detect neonates with acute bilirubin encephalopathy. Other more MRI multimodal images combined with T1-weighted MRI images are expected to use to improve the accuracy in future work. However, this study demonstrates that the semi-quantitative measurement based on T1-weighted MRI images is a simple and compromised way to discriminate acute bilirubin encephalopathy and non-acute bilirubin encephalopathy neonates with hyperbilirubinemia, which may be helpful in improving the current manual diagnosis.

[Color characterization and its correlation with quality index during ripening of Rubus chingii].

The color of Rubus chingii was characterized by digital method, and the content of water extract, alcohol extract, total flavonoids, total polysaccharides, total polyphenols, ellagic acid, linden glycoside, kaophenol-3-O-rutin were determined. Correlation regression was used to analyze the correlation between color and composition. The results showed that L~* was positively correlated with total polyphenols, kaophenol-3-O-rutin and tilide, and moderately positively correlated with total flavones, ellagic acid and aqueous extracts. The a~* value was negatively correlated with total polyphenols, kaophenol-3-O-rutin, and linden glycosides, while was moderately correlated with total flavones, aqueous extracts, and ellagic acid. The b~* value was negatively correlated with the water extract, and moderately correlated with the content of total polyphenols, total polysaccharides, alcohol extract and kaophenol-3-O-rutin, which showed that R. chingii mature color had a significant correlation with material composition in the process of dynamic change. According to the law of dynamic change in the color and quality indexes, it is determined that the appropriate harvest time is in late April to May 1, while the fruit is not turn yellow. The agronomic traits related to fruit was(12.49±0.56) mm in diameter,(14.25±1.19)mm in height,(1.20±0.14) g in weight, the chroma L~* value was 52.87±3.14,a~* value was 2.01±1.58, b~* values was 28.31±3.88. The results lay a foundation for establishing an objective quantitative evaluation model of R. chingii color from experience.

[Clinical effect of multi-oil fat emulsion for parenteral nutrition support in extremely low birth weight infants].

OBJECTIVE: To study the clinical effect of multi-oil fat emulsion for parenteral nutrition support in extremely low birth weight (ELBW) infants. METHODS: A retrospective analysis was performed for 49 ELBW infants who were admitted from January 1, 2018 to July 30, 2020, with an age of ≤14 days on admission and a duration of parenteral nutrition of > 14 days. According to the type of lipid emulsion received, the ELBW infants were divided into two groups: soybean oil, medium-chain triglycerides, olive oil, and fish oil (SMOF) (n=26) and medium-chain triglycerides/long-chain triglycerides (MCT/LCT) (n=23). The two groups were compared in terms of clinical features, complications, nutrition support therapy, and outcome. RESULTS: The 49 ELBW infants had a mean birth weight of (892±83) g and a mean gestational age of (28.2±2.3) weeks. There was no significant difference between the two groups in the incidence rates of hemodynamically significant patent ductus arteriosus, intraventricular hemorrhage, neonatal necrotizing enterocolitis, retinopathy of prematurity, bronchopulmonary dysplasia (BPD), grade Ⅲ BPD, sepsis, and pneumonia (P > 0.05). There was also no significant difference in the duration of parenteral nutrition, the age of total enteral nutrition, and head circumference/body length/body weight at discharge between the two groups (P > 0.05). Of all the infants, 22 (45%) had parenteral nutrition-associated cholestasis (PNAC), with 13 (50%) in the SMOF group and 9 (39%) in the MCT/LCT group but there was no significant difference in the incidence of PNAC between the two groups (P > 0.05); however, the infants with PNAC in the SMOF group had significantly lower peak values of direct bilirubin and alanine aminotransferase than those in the MCT/LCT group (P < 0.05). CONCLUSIONS: The application of multi-oil fat emulsion in ELBW infants does not reduce the incidence rate of complications, but compared with MCT/LCT emulsion, SMOF can reduce the severity of PNAC in ELBW infants.

Hyperinsulinemia rather than insulin resistance itself induces blood pressure elevation in high fat diet-fed rats.

OBJECTIVE: To investigate if insulin resistance per se or the accompanying hyperinsulinemia induced hypertension and its underlying mechanisms. METHODS: Sprague-Dawley rats were randomized into normal diet-fed group (ND group) and high-fat diet-fed group (HFD group). Then, the HFD group was further randomly divided into the control group (HFD_C group), the PIO group (treated with pioglitazone), the STZ_DM group (to induce diabetes with streptozotocin) and the DM+Ins group (streptozotocin injection followed by insulin treatment). Insulin sensitivity, plasma insulin, endothelin-1, norepinephrine, aldosterone, angiotensinⅡ and 24-h urinary sodium excretion (USE) levels of the groups were measured and analyzed. A multiple stepwise regression analysis method was applied to exam our hypothesis. RESULTS: Compared to HFD_C group, the groups with lower plasma insulin, the PIO group and STZ_DM group, showed higher USE and lower blood pressure. The groups with higher plasma insulin (but same level of insulin resistance), the HFD_C group and DM+Ins group, showed lower USE and higher blood pressure. The 24-h urinary sodium excretion was the most important contributor to the significant changes of blood pressure with an R2 of 25.2% in this animal experiment. CONCLUSIONS: It is the compensatory hyperinsulinemia rather than insulin resistance per se that causes blood pressure elevation. The urinary sodium excretion is the key mediator among the multiple mechanisms. Therapies targeting hyperinsulinemia and restricting salt intake may favor a better control of hypertension associated with insulin resistance.

[Effects of different application amounts of potassium fulvic acid on yield and quality of Fritillaria thunbergii].

Fritillaria thunbergii is a commonly used traditional Chinese medicine, which has the effects of clearing heat and resolving stagnation, eliminating phlegm and relieving cough. At present, it is mostly produced by cultivation, and the cultivation process requires application of base fertilizer, winter fertilizer, seedling fertilizer and late top dressing. Now farmyard manure or organic fertilizer can be used to replace the base fertilizer and winter fertilizer, but the research on the replacement of organic fertilizer has not been completed for the late top dressing. Potassium fulvate is a kind of fulvate fertilizer, which can not only regulate the growth of crops but also supplement potassium necessary for the growth of crops. In this paper, using F. thunbergii as a model plant with mature cultivation techniques, the effect of potassium fulvate on the quality and yield of rhizome traditional Chinese medicine F. thunbergii was systematically studied for the first time. HPLC-ELSD was used to determine the contents of peimine A and peimine B, hot dip method was used to determine the content of alcohol extract, and the SPAD-502 Plus chlorophyll meter was used to detect SPAD value. The results showed that applying 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare could effectively improve the yield of F. thunbergii and there was significantly difference between potassium phosphate monobasic and potassium fulvic acid in terms of quality. After the application of range 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare, the content of alcohol soluble extract of F. thunbergii was ranged 21.61% to 22.27%, the total amount of peimine A and peimine B were ranged 0.09% to 0.10%. Applying 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare could replace the conventional pure chemical fertilizer potassium phosphate monobasic, which could be used as top dressing fertilizer for the cultivation of F. thunbergii.

[Application of vacuum stretcher combined with feeding in cranial magnetic resonance imaging examination for neonates: a prospective randomized controlled study].

OBJECTIVE: To study the effect and safety of vacuum stretcher combined with feeding in cranial magnetic resonance imaging (MRI) examination for neonates. METHODS: A prospective study was performed for the neonates with hyperbilirubinemia, with a gestational age of >34 weeks and stable vital signs, who needed cranial MRI examination and did not need oxygen inhalation hospitalized in the Department of Neonatology, Children's Hospital of Zhejiang University School of Medicine, from September to November, 2019. The neonates were randomly divided into a vacuum stretcher combined with feeding group and a conventional sedation group. Vital signs were monitored before, during, and after MRI examination. The success rate of MRI procedure was recorded. RESULTS: A total of 80 neonates were enrolled in the study, with 40 neonates in the vacuum stretcher combined with feeding group and 40 in the conventional sedation group. The vacuum stretcher combined with feeding group had a significantly higher success rate of MRI procedure than the conventional sedation group (P<0.05). As for the neonates who underwent successful MRI examination, the fastest heart rate after examination in the vacuum stretcher combined with feeding group was significantly lower than that in the conventional sedation group (P<0.05), while there were no significant differences between the two groups in transcutaneous oxygen saturation, respiratory rate, and body temperature before and after MRI examination (P>0.05). No complications, such as apnea, acute allergic reactions, and malignant fever, were observed. CONCLUSIONS: Vacuum stretcher combined with feeding can improve the success rate of MRI procedure and reduce the use of sedatives, and meanwhile, it does not increase related risks.