miR171-targeted SCARECROW-LIKE genes CsSCL2 and CsSCL3 regulate somatic embryogenesis in citrus.

Somatic embryogenesis (SE) is a key regeneration pathway in various biotechnology approaches to crop improvement, especially for economically important perennial woody crops like citrus. However, maintenance of SE capability has long been a challenge and becomes a bottleneck in biotechnology-facilitated plant improvement. In the embryogenic callus (EC) of citrus, we identified 2 csi-miR171c-targeted SCARECROW-LIKE genes CsSCL2 and CsSCL3 (CsSCL2/3), which exert positive feedback regulation on csi-miR171c expression. Suppression of CsSCL2 expression by RNA interference (RNAi) enhanced SE in citrus callus. A thioredoxin superfamily protein CsClot was identified as an interactive protein of CsSCL2/3. Overexpression of CsClot disturbed reactive oxygen species (ROS) homeostasis in EC and enhanced SE. Chromatin immunoprecipitation sequencing (ChIP-Seq) and RNA-Seq identified 660 genes directly suppressed by CsSCL2 that were enriched in biological processes including development-related processes, auxin signaling pathway, and cell wall organization. CsSCL2/3 bound to the promoters of regeneration-related genes, such as WUSCHEL-RELATED HOMEOBOX 2 (CsWOX2), CsWOX13, and Lateral Organ Boundaries Domain 40 (LBD40), and repressed their expression. Overall, CsSCL2/3 modulate ROS homeostasis through the interactive protein CsClot and directly suppress the expression of regeneration-related genes, thus regulating SE in citrus. We uncovered a regulatory pathway of miR171c-targeted CsSCL2/3 in SE, which shed light on the mechanism of SE and regeneration capability maintenance in citrus.

Quantitative assessment of normal hip cartilage in children under 9 years old by T2 mapping.

OBJECTIVE: To investigate the variation in T2 at different zones of normal hip cartilage in children and the relationship between T2 value and age. MATERIALS AND METHODS: Nineteen children with 30 normal hip joints were evaluated with a coronal T2 mapping sequence at a 3-Tesla MRI system. The femoral cartilage and acetabular cartilage were firstly segmented by mask-based interactive method and then equally divided into eight and six radial sections, respectively. Moreover, each radial section was further divided into two layers referring to the superficial and deep halves of the corresponding cartilage. Cartilage T2 of these sections and layers were measured and subsequently analyzed. RESULTS: There was a negative correlation between the T2 values in the hip cartilage and the age of children (rs < - 0.6, P1 < 0.05). Articular cartilage T2 increased at angles close to the magic angle (54.7°). Femoral cartilage and acetabular cartilage had a relatively shorter T2 in the radial sections near the vertex of the femoral head. The T2 values in superficial layers of both cartilages were significantly higher than those in deep layers (P < 0.05). CONCLUSION: The T2 value decreases as the cartilage developing into a more mature state. Cartilage T2 values in the weight-bearing areas are relatively low due to an increase of collagen density and the loss of interstitial water. The restriction of the water molecules by solid components in the deeper layer of cartilage may decrease the T2 values.

Multimodel magnetic resonance imaging of mass-forming autoimmune pancreatitis: differential diagnosis with pancreatic ductal adenocarcinoma.

PURPOSE: To assess the value of the multimodel magnetic resonance imaging (MRI), including unenhanced images, dynamic contrast-enhanced MRI (DCE-MRI), MR-cholangiopancreatography (MRCP), and diffusion-weighted imaging (DWI), in differentiation of mass-forming autoimmune pancreatitis (AIP) from pancreatic ductal adenocarcinoma (PDAC). METHODS: Twelve patients with mass-forming AIP and 30 with PDAC were included. All patients underwent unenhanced MRI, DCE-MRI, DWI, and MRCP. Relevant values including sensitivity and specificity of the imaging features and their diagnostic performance for predicting mass-forming AIP were analyzed. RESULTS: Several statistically significant MR findings and quantitative indexes differentiating mass-forming AIP from PDAC, including multiplicity, irregularity or conformation, capsule-like rim enhancement, absence of internal cystic or necrotic portion, homogeneous enhancement during pancreatic, venous, and delayed phases, skipped stricture or stricture of MPD, absence of side branch dilation, maximum upstream MPD diameter < 2.4 mm, ContrastUP > 0.739, ContrastAP > 0.710, ContrastPP > 0.879, and ContrastVP or ContrastDP > 0.949, indicated mass-forming AIP (P < 0.05). The apparent diffusion coefficient (ADC) value was also significantly lower in mass-forming AIP compared to that in PDAC (P = 0.006). The cutoff value of ADC for distinguishing mass-forming AIP from PDAC was 1.099 × 10-3 mm2/s. CONCLUSION: Multimodel MRI, including unenhanced MRI, DCE-MRI with DWI and MRCP can provide qualitative and quantitative information about mass-forming AIP characterization. Multimodel MRI are valuable for differentiating mass-forming AIP from PDAC.

Assessment of irreducible aspects in developmental hip dysplasia by magnetic resonance imaging.

BACKGROUND: The developmental dysplasia of the hip (DDH) can cause a wide range of pathological changes, and often requires surgical treatment. Preoperative evaluation is very important for DDH. We aimed to assess the diagnostic capability of magnetic resonance imaging (MRI) for irreducible aspects preventing hip reduction in DDH. METHODS: A total of 39 pediatric patients who received DDH evaluation in pediatric orthopedics from January 2015 to December 2019 were included. The samples included 4 cases of bilateral DDH and 35 cases of unilateral DDH, a total of 43 hip joint samples. All patients underwent surgical treatment, pathological examination and MRI of hip joint. RESULTS: With pathological results or intraoperative findings as the gold standard, the sensitivity and specificity of MRI were 90.3% and 83.3% for the affected labrum, 92% and 83.3% for thickening of the round ligament, 90.0% and 91.3% for atrophy of the iliopsoas muscle, and 100% and 100% for fibrofatty pulvinar tissue and joint effusion, respectively. CONCLUTIONS: The MRI showed an extraordinary capability of detecting these irreducible factors and helped surgeon choose the appropriate treatment strategies.

Genome-scale mRNA and small RNA transcriptomic insights into initiation of citrus apomixis.

Nucellar embryony (NE) is an adventitious form of apomixis common in citrus, wherein asexual embryos initiate directly from nucellar cells surrounding the embryo sac. NE enables the fixation of desirable agronomic traits and the production of clonal offspring of virus-free rootstock, but impedes progress in hybrid breeding. In spite of the great importance of NE in citrus breeding and commercial production, little is understood about the underlying molecular mechanisms. In this study, the stages of nucellar embryo initiation (NEI) were determined for two polyembryonic citrus cultivars via histological observation. To explore the genes and regulatory pathways involved in NEI, we performed mRNA-seq and sRNA-seq analyses of ovules immediately prior to and at stages during NEI in the two pairs of cultivars. A total of 305 differentially expressed genes (DEGs) were identified between the poly- and monoembryonic ovules. Gene ontology (GO) analysis revealed that several processes are significantly enriched based on DEGs. In particular, response to stress, and especially response to oxidative stress, was over-represented in polyembryonic ovules. Nearly 150 miRNAs, comprising ~90 conserved and ~60 novel miRNAs, were identified in the ovules of either cultivar pair. Only two differentially expressed miRNAs (DEMs) were identified, of which the novel miRN23-5p was repressed whereas the targets accumulated in the polyembryonic ovules. This integrated study on the transcriptional and post-transcriptional regulatory profiles between poly- and monoembryonic citrus ovules provides new insights into the mechanism of NE, which should contribute to revealing the regulatory mechanisms of plant apomixis.

Multicomponent domino reactions of hydrazinecarbodithioates: concise access to 3-substituted 5-thiol-1,3,4-thiadiazolines.

Two classes of addition/cycloaddition cascade reactions of hydrazinecarbodithioate (1) have been developed under mild reaction conditions. Reaction of hydrazinecarbodithioate (1) with formaldehyde solution (2) and propiolic acid (3) gives 3-propargyl-5-thiol-2,3-dihydro-1,3,4-thiadiazoles (5) via a decarboxylative coupling/cycloaddition domino sequence. When propiolic acid (3) is switched to phenyl boronic acid (4), a petasis/cycloaddition domino reaction is instead observed, in which 3-benzyl-5-thiol-2,3-dihydro-1,3,4-thiadiazoles (6) are obtained. Both these reactions show a wide range of functional-group compatibility for propiolic acids and aryl boronic acids, and give the corresponding products in moderate to good yields.

Mild and catalyst-free petasis/decarboxylative domino reaction: chemoselective synthesis of N-benzyl propargylamines.

Multicomponent domino reactions are attractive for assembling functionalized compounds. To this end, a one-pot catalyst-free chemoselective synthesis of N-benzyl propargylamines is reported with good functional group compatibility. This mild process involves in situ formation of an active amine through Petasis reaction of primary amines, formaldehyde solution, and boronic acids, which reacts with propiolic acids to give product in up to 94% yield via decarboxylative coupling reaction.

Efficacy and safety of orlistat in controlling the progression of prediabetes to diabetes: A meta-analysis and systematic review.

BACKGROUND: The aim of this study is to examine the impact of the Orlistat on glucose levels and glucose tolerance in individuals with prediabetes, as well as assess its efficacy and safety in preventing the progression to diabetes. METHODS: For achieving the appropriate randomized controlled trials, we enrolled the public datas from the following electronic databases: The Cochrane library, Embase, China National Knowledge Infrastructure, VIP, Wan-Fang, and China Biology Medicine disc. The article focused on the orlistat intervention of glucose tolerance and glycemic status in prediabetic patients. We restricted the publication time from the creation to May 2023. RESULTS: Six subjects were included in the study, with a total of 1076 participants (532 in the control group vs 544 in the experimental group). The results indicated that the orlistat can reduce the fasting blood glucose [relative risk (RR) = -2.18, 95% confidence intervals (CI) (-2.471, -1.886)], as well as the 2 hour postprandial blood glucose [RR = -1.497, 95% CI (-1.811, -1.183)]. Furthermore, it can prevent the impaired glucose tolerance patients to type 2 diabetes mellitus [RR = 0.605, 95% CI (0.462, 0.791)], and reversal the impaired glucose tolerance [RR = 2.092, 95% CI (1.249, 3.503)]. CONCLUSIONS: In prediabetic people, the orlistat can control weight, reduce the fasting blood glucose and the 2 hour postprandial blood glucose, and then delay the progression of diabetes. However, due to the quantitative restrictions, additional high-quality study needs to be conducted to improve the reliability of the results.

Microenvironment-induced CREPT expression by cancer-derived small extracellular vesicles primes field cancerization.

Rationale: Cancer local recurrence increases the mortality of patients, and might be caused by field cancerization, a pre-malignant alteration of normal epithelial cells. It has been suggested that cancer-derived small extracellular vesicles (CDEs) may contribute to field cancerization, but the underlying mechanisms remain poorly understood. In this study, we aim to identify the key regulatory factors within recipient cells under the instigation of CDEs. Methods: In vitro experiments were performed to demonstrate that CDEs promote the expression of CREPT in normal epithelial cells. TMT-based quantitative mass spectrometry was employed to investigate the proteomic differences between normal cells and tumor cells. Loss-of-function approaches by CRISPR-Cas9 system were used to assess the role of CREPT in CDEs-induced field cancerization. RNA-seq was performed to explore the genes regulated by CREPT during field cancerization. Results: CDEs promote field cancerization by inducing the expression of CREPT in non-malignant epithelial cells through activating the ERK signaling pathway. Intriguingly, CDEs failed to induce field cancerization when CREPT was deleted, highlighting the importance of CREPT. Transcriptomic analyses revealed that CDEs elicited inflammatory responses, primarily through activation of the TNF signaling pathway. CREPT, in turn, regulates the transduction of downstream signals of TNF by modulating the expression of TNFR2 and PI3K, thereby promoting inflammation-to-cancer transition. Conclusion: CREPT not only serves as a biomarker for field cancerization, but also emerges as a target for preventing the cancer local recurrence.

Combined diagnosis of multiparametric MRI-based deep learning models facilitates differentiating triple-negative breast cancer from fibroadenoma magnetic resonance BI-RADS 4 lesions.

PURPOSE: To investigate the value of the combined diagnosis of multiparametric MRI-based deep learning models to differentiate triple-negative breast cancer (TNBC) from fibroadenoma magnetic resonance Breast Imaging-Reporting and Data System category 4 (BI-RADS 4) lesions and to evaluate whether the combined diagnosis of these models could improve the diagnostic performance of radiologists. METHODS: A total of 319 female patients with 319 pathologically confirmed BI-RADS 4 lesions were randomly divided into training, validation, and testing sets in this retrospective study. The three models were established based on contrast-enhanced T1-weighted imaging, diffusion-weighted imaging, and T2-weighted imaging using the training and validation sets. The artificial intelligence (AI) combination score was calculated according to the results of three models. The diagnostic performances of four radiologists with and without AI assistance were compared with the AI combination score on the testing set. The area under the curve (AUC), sensitivity, specificity, accuracy, and weighted kappa value were calculated to assess the performance. RESULTS: The AI combination score yielded an excellent performance (AUC = 0.944) on the testing set. With AI assistance, the AUC for the diagnosis of junior radiologist 1 (JR1) increased from 0.833 to 0.885, and that for JR2 increased from 0.823 to 0.876. The AUCs of senior radiologist 1 (SR1) and SR2 slightly increased from 0.901 and 0.950 to 0.925 and 0.975 after AI assistance, respectively. CONCLUSION: Combined diagnosis of multiparametric MRI-based deep learning models to differentiate TNBC from fibroadenoma magnetic resonance BI-RADS 4 lesions can achieve comparable performance to that of SRs and improve the diagnostic performance of JRs.

Genome-wide identification of the C2H2-Zinc finger gene family and functional validation of CsZFP7 in citrus nucellar embryogenesis.

KEY MESSAGE: Genome-wide identification of C2H2-ZF gene family in the poly- and mono-embryonic citrus species and validation of the positive role of CsZFP7 in sporophytic apomixis. The C2H2 zinc finger (C2H2-ZF) gene family is involved in plant vegetative and reproductive development. Although a large number of C2H2 zinc-finger proteins (C2H2-ZFPs) have been well characterized in some horticultural plants, little is known about the C2H2-ZFPs and their function in citrus. In this work, we performed a genome-wide sequence analysis and identified 97 and 101 putative C2H2-ZF gene family members in the genomes of sweet orange (C. sinensis, poly-embryonic) and pummelo (C. grandis, mono-embryonic), respectively. Phylogenetic analysis categorized citrus C2H2-ZF gene family into four clades, and their possible functions were inferred. According to the numerous regulatory elements on promoter, citrus C2H2-ZFPs can be divided into five different regulatory function types that indicate functional differentiation. RNA-seq data revealed 20 differentially expressed C2H2-ZF genes between poly-embryonic and mono-embryonic ovules at two stages of citrus nucellar embryogenesis, among them CsZFP52 specifically expressed in mono-embryonic pummelo ovules, while CsZFP7, 37, 44, 45, 67 and 68 specifically expressed in poly-embryonic sweet orange ovules. RT-qPCR further validated that CsZFP7 specifically expressed at higher levels in poly-embryonic ovules, and down-regulation of CsZFP7 in the poly-embryonic mini citrus (Fortunella hindsii) increased rate of mono-embryonic seeds compared with the wild type, indicating the regulatory potential of CsZFP7 in nucellar embryogenesis of citrus. This work provided a comprehensive analysis of C2H2-ZF gene family in citrus, including genome organization and gene structure, phylogenetic relationships, gene duplications, possible cis-elements on promoter regions and expression profiles, especially in the poly- and mono-embryogenic ovules, and suggested that CsZFP7 is involved in nucellar embryogenesis.

Noninvasive assessment of breast cancer molecular subtypes on multiparametric MRI using convolutional neural network with transfer learning.

BACKGROUND: To evaluate the performances of multiparametric MRI-based convolutional neural networks (CNNs) for the preoperative assessment of breast cancer molecular subtypes. METHODS: A total of 136 patients with 136 pathologically confirmed invasive breast cancers were randomly divided into training, validation, and testing sets in this retrospective study. The CNN models were established based on contrast-enhanced T1 -weighted imaging (T1 C), Apparent diffusion coefficient (ADC), and T2 -weighted imaging (T2 W) using the training and validation sets. The performances of CNN models were evaluated on the testing set. The area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and accuracy were calculated to assess the performance. RESULTS: For the separation of each subtype from other subtypes on the testing set, the T1 C-based models yielded AUCs from 0.762 to 0.920; the ADC-based models yielded AUCs from 0.686 to 0.851; and the T2 W-based models achieved AUCs from 0.639 to 0.697. CONCLUSION: T1 C-based models performed better than ADC-based models and T2 W-based models in assessing the breast cancer molecular subtypes. The discriminating performances of our CNN models for triple negative and human epidermal growth factor receptor 2-enriched subtypes were better than that of luminal A and luminal B subtypes.

Fabrication and characterization of the H/J-type aggregates astaxanthin/bovine serum albumin/chitosan nanoparticles.

Astaxanthin is a natural liposoluble ketocarotenoid with various biological activities. Hydrophobic astaxanthin with C2h symmetry can self-assembly form H-type aggregates and J-type aggregates in hydrated polar solvents. However, astaxanthin and its aggregates are limited by its water insolubility and chemical instability. Here, the biological macromolecules bovine serum albumin (BSA) and chitosan were chosen as protein-polysaccharides based delivery systems for astaxanthin aggregates by molecular self-assembly method. The precise prepared H-ABC-NPs and J-ABC-NPs suspensions were both near spheres with hydrodynamic size around 281 ± 9 nm and 368 ± 5 nm and zeta potentials around +26 mV and +30 mV, respectively. Two types of astaxanthin aggregates were distinguished, water-dispersible, and stable in nanocarriers through UV-vis spectra observation. The encapsulation efficiency of the astaxanthin in ABC-NPs was above 90 %. Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD) analyses indicated that the dominant driving forces of ABC-NPs formation mainly included electrostatic, hydrophobic interactions and hydrogen bonding. These results offer an elegant opportunity for the protein-polysaccharides delivery systems, and provide an important perspective for applying novel water-dispersed astaxanthin aggregates products in nutrition and medicine industry.

Apomixis: genetic basis and controlling genes.

Apomixis is the phenomenon of clonal reproduction by seed. As apomixis can produce clonal progeny with exactly the same genotype as the maternal plant, it has an important application in genotype fixation and accelerating agricultural breeding strategies. The introduction of apomixis to major crops would bring many benefits to agriculture, including permanent fixation of superior genotypes and simplifying the procedures of hybrid seed production, as well as purification and rejuvenation of crops propagated vegetatively. Although apomixis naturally occurs in more than 400 plant species, it is rare among the major crops. Currently, with better understanding of apomixis, some achievements have been made in synthetic apomixis. However, due to prevailing limitations, there is still a long way to go to achieve large-scale application of apomixis to crop breeding. Here, we compare the developmental features of apomixis and sexual plant reproduction and review the recent identification of apomixis genes, transposons, epigenetic regulation, and genetic events leading to apomixis. We also summarize the possible strategies and potential genes for engineering apomixis into crop plants.

Umbilical cord mesenchymal stem cell-derived exosomes alleviate collagen-induced arthritis by balancing the population of Th17 and regulatory T cells.

Exosomes released by mesenchymal stem cells (MSCs) are thought to function as extensions of the MSCs. However, it remains unclear whether exosomes derived from human umbilical cord MSCs (HUMSCs) possess immunoregulatory functions in rheumatoid arthritis. We report that when mice with collagen-induced arthritis were injected with exosomes derived from HUMSC (HUMSC-Exo), their paws became less swollen, and they had lower serum pro-inflammatory cytokine and anti-collagen IgG levels, and decreased synovial hyperplasia. The HUMSC-Exo appeared to restore the balance between Th17 and Treg cells, and this effect was accompanied by reduced IL-17 and enhanced TGF-ß and IL-10 levels. These findings suggest that HUMSC-Exo function as important regulator of the balance between Th1/Th17 and Treg cells during immune and inflammatory responses.

Magnetic resonance imaging investigation of age-related morphological changes in the pancreases of 226 Chinese.

OBJECTIVE: To investigate the morphological changes with age in the pancreases of healthy individuals undergoing magnetic resonance imaging (MRI). METHODS: The participants were selected from adults who were undergoing physical examinations from January 2017 to September 2020 at Huadong Hospital. They were divided according to age, as broken down by decades into seven groups ranging from 20 to 29 years to ≥80 years of age. There were 30 to 35 cases for each decade. They were then divided into a young and middle-aged group (<60 years of age) and an elderly group (≥60 years of age). The morphological characteristics of the pancreases of each participant in the group were measured on magnetic resonance images. The characteristics included the pancreatic anteroposterior diameters and volumes. The relationships between the anteroposterior diameters of the pancreatic head, body, and tail and pancreatic volume and age were analyzed. RESULTS: A total of 226 magnetic resonance images from 112 (49.56%) men and 114 (50.44%) women, aged 22-93 (54.68 ± 19.52) years. The age ranges of the seven groups consisted of the following: 20-29 years (n = 33), 30-39 years (n = 32), 40-49 years (n = 32), 50-59 years (n = 31), 60-69 years (n = 35), 70-79 years (n = 33) and ≥80 years (n = 30). The age range and numbers of patients in the young and middle-aged group was 22-59 (40.09 ± 10.88) years (n = 128) and in the elderly group was 60-93 (73.74 ± 8.99) years (n = 98). The MRI findings characteristic of aging included pancreatic atrophy (especially of the pancreatic tail), pancreatic lobulation, uneven signal intensity, fatty degeneration, and widening of the main pancreatic duct. The respective anteroposterior diameters of the pancreatic head, body, and tail and the pancreatic volumes peaked at 30 to 39 years as follows: 28.03 ± 4.45 mm, 24.10 ± 4.27 mm, 24.57 ± 4.94 mm, 98.54 ± 26.56 cm3; and then gradually decreased to 19.05 ± 3.59 mm, 16.00 ± 3.81 mm, 13.83 ± 3.39 mm, 45.02 ± 9.15 cm3 at ≥80 years, for respective decreases of 32.03%, 33.60%, 43.71%, and 54.31%. The respective anteroposterior diameters of the pancreatic head, body, tail, and pancreatic volume in the elderly patients were 21.45 ± 4.15 mm, 18.14 ± 4.09 mm, 16.81 ± 4.37 mm, and 59.02 ± 21.44 cm3, which were significantly smaller than the respective corresponding measurements in the young and middle-aged patients (26.09 ± 4.40 mm, 22.30 ± 4.42 mm, 22.08 ± 4.53 mm, and 88.32 ± 23.92 cm3). The differences were statistically significant (t = 8.06, 7.24, 8.79, 9.54, respectively, p < 0.001). The anteroposterior diameters of the pancreatic head, body, tail, and pancreatic volume were negatively correlated with age (r = -0.53, -0.47, -0.56, -0.57, respectively, p < 0.001). CONCLUSION: The anteroposterior diameters of the pancreatic head, body, tail, and the pancreatic volume all peaked at the age range of 30-39 years and then gradually decreased with increasing age. After the age of 60 years, pancreatic atrophy became increasingly obvious, with changes in shape and widening with age of the main pancreatic duct.

Regulation of nucellar embryony, a mode of sporophytic apomixis in Citrus resembling somatic embryogenesis.

Apomixis, is an asexual mode of seed formation resulting in genetically identical or clonal seed with a maternal genotype. Apomixis has not been reported in seed crops where its flexible application in plant breeding could accelerate delivery of new varieties. By contrast, a sporophytic form of apomixis termed nucellar or adventitious embryony is common in the Rutaceae containing Citrus crop species. Here, multiple embryos develop from the maternal, somatic, nucellar cells of the ovule. They are incorporated into the enlarging embryo sac containing the sexually derived zygotic embryo and endosperm, which are products of double fertilization. Recent research has provided insights to the molecular basis for nucellar embryony. Here, we review the current understanding of the initiation, genetic basis and evolution of nucellar embryony in Citrus, and discuss prospects for future study and breeding applications of Citrus sporophytic apomixis.

Different characteristics of infants diagnosed with congenital choledochal malformation prenatally or postnatally.

The general condition, clinical and pathological characteristics, and treatment regimens of patients prenatally and postnatally diagnosed with congenital choledochal malformation (CM) were analyzed in order to investigate the clinical significance of early diagnosis, treatment, and intervention in CM. We retrospectively analyzed 33 children who were admitted to the Children's Hospital of Soochow University between 1 March 2010 and 31 May 2019, and their diagnosis of CM was confirmed by radiological, surgical and pathological findings. All the patients were under 36 months of age. The patients were divided into prenatally diagnosed and postnatally diagnosed groups. There were 16 and 17 CM patients in the prenatally and postnatally diagnosed groups, respectively, with a preponderance of females in both groups. Compared with the prenatally diagnosed group, the postnatally diagnosed group had a higher incidence of abdominal pain and vomiting (p < 0.05) and higher AST, GGT, and TB levels (p < 0.05). Although postoperative histopathological examination showed inflammation in both groups, congestion in the cyst walls and fibrous tissue hyperplasia were more significant in the postnatally diagnosed group (p < 0.05). In addition, operation time, length of time required to resume a normal diet after surgery, and total length of hospitalization differed between the 2 groups (p < 0.05), with the prenatally diagnosed group having a relatively longer operation time and taking longer to resume a normal diet after surgery. However, the total length of hospitalization in the prenatally diagnosed group was shorter than that in the postnatally diagnosed group. Compared with prenatally diagnosed CM patients, more symptoms, greater severity of symptoms, and more time to recovery after surgery were observed in postnatally diagnosed CM patients.

Genome of a citrus rootstock and global DNA demethylation caused by heterografting.

Grafting is an ancient technique used for plant propagation and improvement in horticultural crops for at least 1,500 years. Citrus plants, with a seed-to-seed cycle of 5-15 years, are among the fruit crops that were probably domesticated by grafting. Poncirus trifoliata, a widely used citrus rootstock, can promote early flowering, strengthen stress tolerance, and improve fruit quality via scion-rootstock interactions. Here, we report its genome assembly using PacBio sequencing. We obtained a final genome of 303 Mb with a contig N50 size of 1.17 Mb and annotated 25,680 protein-coding genes. DNA methylome and transcriptome analyses indicated that the strong adaptability of P. trifoliata is likely attributable to its special epigenetic modification and expression pattern of resistance-related genes. Heterografting by using sweet orange as scion and P. trifoliata as rootstock and autografting using sweet orange as both scion and rootstock were performed to investigate the genetic effects of the rootstock. Single-base methylome analysis indicated that P. trifoliata as a rootstock caused DNA demethylation and a reduction in 24-nt small RNAs (sRNAs) in scions compared to the level observed with autografting, implying the involvement of sRNA-mediated graft-transmissible epigenetic modifications in citrus grafting. Taken together, the assembled genome for the citrus rootstock and the analysis of graft-induced epigenetic modifications provide global insights into the genetic effects of rootstock-scion interactions and grafting biology.

Transcriptomes and DNA methylomes in apomictic cells delineate nucellar embryogenesis initiation in citrus.

Citrus nucellar poly-embryony (NPE) is a mode of sporophytic apomixis that asexual embryos formed in the seed through adventitious embryogenesis from the somatic nucellar cells. NPE allows clonal propagation of rootstocks, but it impedes citrus cross breeding. To understand the cellular processes involved in NPE initiation, we profiled the transcriptomes and DNA methylomes in laser microdissection captured citrus apomictic cells. In apomictic cells, ribosome biogenesis and protein degradation were activated, whereas auxin polar transport was repressed. Reactive oxygen species (ROS) accumulated in the poly-embryonic ovules, and response to oxidative stress was provoked. The global DNA methylation level, especially that of CHH context, was decreased, whereas the methylation level of the NPE-controlling key gene CitRWP was increased. A C2H2 domain-containing transcription factor gene and CitRWP co-expressed specifically in apomictic cells may coordinate to initiate NPE. The activated embryogenic development and callose deposition processes indicated embryogenic fate of nucellar embryo initial (NEI) cells. In our working model for citrus NPE initiation, DNA hyper-methylation may activate transcription of CitRWP, which increases C2H2 expression and ROS accumulation, triggers epigenetic regulation and regulates cell fate transition and NEI cell identity in the apomictic cells.