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Abstract Background Anthracnose, mainly caused by Colletotrichum fructicola, leads to severe losses in pear production. However, there is limited information available regarding the molecular response to anthracnose in pears. Results In this study, the anthracnose-resistant variety ‘Seli’ and susceptible pear cultivar ‘Cuiguan’ were subjected to transcriptome analysis following C. fructicola inoculation at 6 and 24 h using RNA sequencing. A total of 3186 differentially expressed genes were detected in ‘Seli’ and ‘Cuiguan’ using Illumina sequencing technology. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that the transcriptional response of pears to C. fructicola infection included responses to reactive oxygen species, phytohormone signaling, phenylpropanoid biosynthesis, and secondary metabolite biosynthetic processes. Moreover, the mitogen-activated protein kinase (MAPK) signaling pathway and phenylpropanoid biosynthesis were involved in the defense of ‘Seli’. Furthermore, the gene coexpression network data showed that genes related to plant–pathogen interactions were associated with C. fructicola resistance in ‘Seli’ at the early stage. Conclusion Our results showed that the activation of specific genes in MAPK, calcium signaling pathways and phenylpropanoid biosynthesis was highly related to C. fructicola resistance in ‘Seli’ and providing several potential candidate genes for breeding anthracnose-resistant pear varieties.

Fruitlet calyx shedding in pear plants is apparently regulated via numerous pathways that involve both environmental triggers and phytohormones cues such as auxin. In this study, we found at 10 days after full bloom (DAFB) higher levels of indoleacetic acid (IAA) and tryptophan (Trp) in calyx persistence fruitlet (CPF) than calyx shedding fruitlet (CSF) of ‘Danshan Suli’ pear (Pyrus bretschneideri Rhed.). Consisting with this, the activity of indolealdehyde oxidase (IAAIdO), which promotes IAA synthesis, was remarkably increased, and that of peroxidase (POD), which degrades IAA, dropped markedly in CPF but not in CSF. Further, qRT-PCR results revealed that most of 31 PbrARFs (encoding auxin response factors) in Pyrus bretschneideri were highly expressed in CPF, whereas PbrARF4, PbrARF24 and PbrARF26 were significantly down-regulated in CPF vis-a-vis CSF. Phylogenetic analysis revealed that 6 PbrARFs clustered in the group Ⅲ, where PbrARF4 showed the closest affinity with AtARF1 that promotes organ abscission, indicating a putative role of PbrARF4 in mediating the process of calyx shedding in pear. In fact, the ectopic overexpression of PbrARF4 in Solanum lycopersicum resulted in an earlier-formed and deeper abscission layer (AL) in the transgenic plants, whose calyxes were more prone to wilt at the mature red stage (MR) compared with the control plants (wild-type). More importantly, expression levels of the abscission genes SlLS and SlCel2 in transgenic plants overexpressing PbrARF4 were significantly up-regulated in comparation with the WT, whereas those of SlBI and SlTAPG2 were considerably inhibited. Further, PbrJOINTLESS and PbrIDA, the two genes related to calyx shedding in pear, were up-regulated more in CSF than CPF. The findings contribute to a better understanding of PbrARFs involved in fruitlet calyx shedding of pear, which could prove beneficial to improving the quality of pear fruit.

Abstract Background Chlorophyll (Chl) is an agronomic trait associated with photosynthesis and yield. Gibberellin 2-oxidases (GA2oxs) have previously been shown to be involved in Chl accumulation. However, whether and how the PbrGA2ox proteins (PbrGA2oxs) mediate Chl accumulation in pear (Pyrus spp.) is scarce. Results Here, we aimed to elucidate the role of the pear GA2ox gene family in Chl accumulation and the related underlying mechanisms. We isolated 13 PbrGA2ox genes (PbrGA2oxs) from the pear database and identified PbrGA2ox1 as a potential regulator of Chl accumulation. We found that transiently overexpressing PbrGA2ox1 in chlorotic pear leaves led to Chl accumulation, and PbrGA2ox1 silencing in normal pear leaves led to Chl degradation, as evident by the regreening and chlorosis phenomenon, respectively. Meanwhile, PbrGA2ox1-overexpressing (OE) tobacco plants discernably exhibited Chl built-up, as evidenced by significantly higher Pn and Fv/Fm. In addition, RNA sequencing (RNA-seq), physiological and biochemical investigations revealed an increase in abscisic acid (ABA), methyl jasmonate (MeJA), and salicylic acid (SA) concentrations and signaling pathways; a marked elevation in reducing and soluble sugar contents; and a marginal decline in the starch and sucrose levels in OE plants. Interestingly, PbrGA2ox1 overexpression did not prominently affect Chl synthesis. However, it indeed facilitated chloroplast development by increasing chloroplast number per cell and compacting the thylakoid granum stacks. These findings might jointly contribute to Chl accumulation in OE plants. Conclusion Overall, our results suggested that GA2oxs accelerate Chl accumulation by stimulating chloroplast development and proved the potential of PbrGA2ox1 as a candidate gene for genetically breeding biofortified pear plants with a higher yield.

Abstract Background The primary management modalities for the patella in TKA include patellar resurfacing, patellar non-resurfacing, patellar resurfacing with denervation, and patellar non-resurfacing with denervation. Traditionally, meta-analyses have predominantly focused on examining comparisons between two management modalities. However, this study performed a network meta-analysis to compare all four patellar management interventions to identify the most optimal approach for patellar management in TKA. Methods A computer-based search of PubMed, China National Knowledge Infrastructure (CNKI), The Cochrane Library, Web of science, Embase, and MEDLINE databases was performed to identify randomized controlled trials focusing on the four management interventions for the patella in TKA. Comparisons included two-by-two comparisons as well as those involving more than two concurrent comparisons. The search timeframe spanned from inception to June 30, 2023. Two independent authors extracted the data and evaluated the quality of the literature. The Cochrane Collaboration Risk of Bias (ROB) tool was used to evaluate the overall quality of the literature. Subsequently, a network meta-analysis was conducted using the “gemtc” package of the R-4.2.3 software. Outcome measures such as anterior knee pain (AKP), reoperation rate, and patient satisfaction rate were evaluated using odd ratio (OR) and 95% confidence intervals (CI). Additionally, the knee society score (KSS), function score (FS), and range of motion (ROM) were evaluated using mean differences (MD) with associated 95% CI. The different treatment measures were ranked using the surfaces under the cumulative ranking curves (SUCRA). Results A total of 50 randomized controlled trials involving 9,283 patients were included in the analysis. The findings from this network meta-analysis revealed that patellar resurfacing exhibited significantly lower postoperative reoperation rate (OR 0.44, 95% CI 0.24–0.63) and AKP (OR 0.58, 95% CI 0.32–1) compared to non-resurfacing. Additionally, patellar resurfacing exhibited higher postoperative KSS clinical scores in comparison with non-resurfacing (MD: 1.13, 95% CI 0.18–2.11). However, for postoperative FS, ROM, and patient satisfaction, no significant differences were observed among the four management interventions. Conclusion Patellar resurfacing emerges as the optimal management modality in primary TKA. However, future studies should aim to reduce sources of heterogeneity and minimize the influence of confounding factors on outcomes. Systematic review registration https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42023434418 identifier: CRD42023434418

‘Akizuki’ pear (Pyrus pyrifolia Nakai) corky disease is a physiological disease that strongly affects the fruit quality of ‘Akizuki’ pear and its economic value. In this study, Raman spectroscopy was employed to develop an early diagnosis model by integrating support vector machine (SVM), random forest (RF), gradient boosting decision tree (GBDT), extreme gradient boosting (XGBoost), and convolutional neural network (CNN) modeling techniques. The effects of various pretreatment methods and combinations of methods on modeling results were studied. The relative optimal index formula was utilized to identify the SG and SG+WT as the most effective preprocessing methods. Following the optimal preprocessing method, the performance of the majority of the models was markedly enhanced through the process of model reconditioning, among which XGBoost achieved 80% accuracy under SG+WT pretreatment, and F1 and kappa both performed best. The results show that RF, GBDT, and XGBoost are more sensitive to the pretreatment method, whereas SVM and CNN are more dependent on internal parameter tuning. The results of this study indicate that the early detection of Raman spectroscopy represents a novel approach for the nondestructive identification of asymptomatic ‘Akizuki’ pear corky disease, which is of paramount importance for the realization of large-scale detection across orchards.

Summary: Cardiac dysfunction, an early complication of endotoxemia, is the major cause of death in intensive care units. No specific therapy is available at present for this cardiac dysfunction. Here, we show that the N-terminal gasdermin D (GSDMD-N) initiates mitochondrial apoptotic pore and cardiac dysfunction by directly interacting with cardiolipin oxidized by complex II-generated reactive oxygen species (ROS) during endotoxemia. Caspase-4/11 initiates GSDMD-N pores that are subsequently amplified by the upregulation and activation of NLRP3 inflammation through further generation of ROS. GSDMD-N pores form prior to BAX and VDAC1 apoptotic pores and further incorporate into BAX and VDAC1 oligomers within mitochondria membranes to exacerbate the apoptotic process. Our findings identify oxidized cardiolipin as the definitive target of GSDMD-N in mitochondria of cardiomyocytes during endotoxin-induced myocardial dysfunction (EIMD), and modulation of cardiolipin oxidation could be a therapeutic target early in the disease process to prevent EIMD.

Brown spot, a serious disease caused by Nothophoma quercina, results in small, brown, round-to-oval lesions on the leaves and is a major concern in the pear industry. We previously reported a new pathogenic strain NqHH of Nothophoma quercina, which was isolated from ‘Huanghua’ pear (Pyrus pyrifolia Nakai, cultivar Huanghua) leaves. However, genome information regarding this pathogen is limited. Herein, we detected the genome-wide sequence of the NqHH strain using the Illumina NovaSeq 6000 and PacBio Sequel Ⅱ systems. The genome of NqHH was estimated to be 35.3 Mb with an N50 length of 1.37 Mb, encoding for 11,659 predicted protein-coding genes. This genome sequence provides valuable information for the selection of pathogenic effectors and comparative genomics of brown spot disease in pears. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Background The dried bark of Ailanthus altissima (Mill.) Swingle is widely used in traditional Chinese medicine for the treatment of ulcerative colitis. The objective of this study was to explore the therapeutic basis of the dried bark of Ailanthus altissima (Mill.) Swingle for the treatment of ulcerative colitis based on Virtual Screening–Molecular Docking–Activity Evaluation technology. Methods By searching the Traditional Chinese Medicine Systems Pharmacology TCMSP Database and Analysis Platform, 89 compounds were obtained from the chemical components of the dried bark of Ailanthus altissima (Mill.) Swingle. Then, after preliminarily screening the compounds based on Lipinski’s rule of five and other relevant conditions, the AutoDock Vina molecular docking software was used to evaluate the affinity of the compounds to ulcerative colitis-related target proteins and their binding modes through use of the scoring function to identify the best candidate compounds. Further verification of the compound’s properties was achieved through in vitro experiments. Results Twenty-two compounds obtained from the secondary screening were molecularly docked with ulcerative colitis-related target proteins (IL-1R, TLR, EGFR, TGFR, and Wnt) using AutoDock Vina. The free energies of the highest scoring compounds binding to the active cavity of human IL-1R, TLR, EGFR, TGFR, and Wnt proteins were − 8.7, − 8.0, − 9.2, − 7.7, and − 8.5 kcal/mol, respectively. The potential compounds, dehydrocrebanine, ailanthone, and kaempferol, were obtained through scoring function and docking mode analysis. Furthermore, the potential compound ailanthone (1, 3, and 10 µM) was found to have no significant effect on cell proliferation, though at 10 µM it reduced the level of pro-inflammatory factors caused by lipopolysaccharide. Conclusion Among the active components of the dried bark of Ailanthus altissima (Mill.) Swingle, ailanthone plays a major role in its anti-inflammatory properties. The present study shows that ailanthone has advantages in cell proliferation and in inhibiting of inflammation, but further animal research is needed to confirm its pharmaceutical potential.

AIM: To explore the established method of the diabetic mouse blood glucose control model and preliminary observation of its influence on the retinas of diabetic mice. METHODS: The db/db BKS-DB (Leprko/ko) mice were randomly divided into two groups: the poor blood glucose control group (PG group, n=18) and the stable blood glucose control group (SG group, n=12), with BKS-DB (Leprwt/wt) as the normal blood glucose control group (NG group, n=18). According to the blood glucose values for 5 intervals which were monitored during the period of adaption, the PG group was injected with insulin aspart twice daily, fasted for 2h and then returned to normal. The SG group was injected with insulin aspart twice and insulin glargine once daily and fed with a quantitative ration. Fundus images were collected after eight weeks. The glycosylated hemoglobin (HbA1c), mean blood glucose level (MBG), standard deviation of blood glucose (SDBG), coefficient of variation of blood glucose (CVBG), and mean amplitude of glycemic excursion (MAGE) in each group were examined and calculated. RESULTS: The HbA1c, MBG, SDBG, CVBG, and MAGE levels in the PG group were significantly higher than those in the NG and SG groups (all P0.05). Preliminary observation of fundus images in the PG group and SG groups showed scattered retinal bleeding spots, while bleeding was more obvious in the PG group. CONCLUSION: The blood glucose control model of type 2 diabetes mellitus mice can be successfully established by subcutaneous injection of insulin aspart insulin glargine and rationed food, which is valuable for studying the mechanism of blood glucose fluctuations in diabetic complications in vivo.

Abstract COVID-19 raises attention to epidemic transmission in various places. This study analyzes the transmission risks associated with human activity places at multiple scales, including different types of settlements and eleven types of specific establishments (restaurants, bars, etc.), using COVID-19 data in 906 urban areas across four continents. Through a difference-in-difference approach, we identify the causal effects of activities at various places on epidemic transmission. We find that at the micro-scale, though the transmission risks at different establishments differ across countries, sports, entertainment, and catering establishments are generally more infectious. At the macro-scale, contradicting common beliefs, it is consistent across countries that transmission does not increase with settlement size and density. It is also consistent that specific establishments play a lesser role in transmission in larger settlements, suggesting more transmission happening elsewhere. These findings contribute to building a system of knowledge on the linkage between places, human activities, and disease transmission.

Background: Traditional full-incision double eyelid blepharoplasty has several significant disadvantages, including slow recovery and bilateral asymmetry, which results in high rates of patient dissatisfaction. These problems can be minimized using improved surgical techniques aimed at reducing tissue damage and bleeding, decreasing operation time, and especially at protecting the subcutaneous capillary network during the operation. We therefore aimed to guard the capillary network and reduce intraoperative bleeding using the practical technique of reverse orbicularis oculi muscle resection. Methods: Fifty cases of double eyelid blepharoplasty with traditional full-incision (group A) and 61 cases of reverse double eyelid blepharoplasty with a preserved capillary network (group B) were divided into two groups from October 1 to December 31, 2021 and from March 1 to May 31, 2022, respectively. The orbicularis oculi muscle was removed reversely from the front of the tarsal plate using double eyelid blepharoplasty, and the muscular membrane and subcutaneous capillary network were preserved. The operation time recorded in previous medical records was checked and compared with that of our novel technique. The recovery of double eyelid blepharoplasty was determined at 1, 4, and 12 weeks after the operation and patient satisfaction was also compared with that in previous medical records of patients in group A. Results: The reverse double eyelid blepharoplasty procedure was less complicated and easier to control than the traditional full-incision double eyelid blepharoplasty methodology. The average operation time of the double eyelid blepharoplasty cases with traditional full-incision was significantly greater than that of the cases of reverse double eyelid blepharoplasty with the capillary network preserved (45.68 ​± ​5.27 ​min and 31.11 ​± ​5.58 ​min, respectively; t=14.018 9, P=0.000 0). Patient satisfaction was 42% and 78.69%, respectively, at 1 week post-operation (χ2=15.724 8, P=0.000 1), 62% and 90.16% at 4 weeks post-operation (χ2=12.490 2, P=0.000 4), and 90% and 95.08%, at 12 weeks post-operation (χ2=1.061 1, P=0.303 0). Conclusion: Reverse double eyelid blepharoplasty with preserved subcutaneous capillary network effectively decreases the operation time and improves post-operative patient satisfaction. It is a double eyelid blepharoplasty method worthy of popularization.

Drought and high-temperature stress are the main abiotic stresses that alone or simultaneously affect the yield and quality of pears worldwide. However, studies on the mechanisms of drought or high-temperature resistance in pears remain elusive. Therefore, the molecular responses of Pyrus betuleafolia, the widely used rootstock in pear production, to drought and high temperatures require further study. Here, drought- or high-temperature-resistant seedlings were selected from many Pyrus betuleafolia seedlings. The leaf samples collected before and after drought or high-temperature treatment were used to perform RNA sequencing analysis. For drought treatment, a total of 11,731 differentially expressed genes (DEGs) were identified, including 4444 drought-induced genes and 7287 drought-inhibited genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these DEGs were more significantly enriched in plant hormone signal transduction, flavonoid biosynthesis, and glutathione metabolism. For high-temperature treatment, 9639 DEGs were identified, including 5493 significantly upregulated genes and 4146 significantly downregulated genes due to high-temperature stress. KEGG analysis showed that brassinosteroid biosynthesis, arginine metabolism, and proline metabolism were the most enriched pathways for high-temperature response. Meanwhile, the common genes that respond to both drought and high-temperature stress were subsequently identified, with a focus on responsive transcription factors, such as MYB, HSF, bZIP, and WRKY. These results reveal potential genes that function in drought or high-temperature resistance. This study provides a theoretical basis and gene resources for the genetic improvement and molecular breeding of pears.

Cold stress is a prominent abiotic factor that adversely affects the growth and yield of pears, consequently restricting the cultivation range and resulting in substantial economic losses for the pear industry. Basic region–leucine zipper (bZIP) transcription factors are widely involved in multiple physiological and biochemical activities of plants, particularly in response to cold stress. In this study, the responsiveness of PbbZIP11 in pear to cold stress was investigated, and its role was explored by using pear callus and Arabidopsis thaliana. The findings revealed that overexpression of PbbZIP11 enhanced the tolerance of pear callus and Arabidopsis thaliana to cold stress. The antioxidant enzyme activities of transgenic plants were enhanced and the expression of C-repeat binding transcription factor (CBF) genes was increased as compared to wild-type plants. To better understand the biological function of PbbZIP11, mRNAs were isolated from overexpressed and wild-type Arabidopsis thaliana after cold stress for whole-genome sequencing. The results showed that the expression of some CBF downstream target genes changed after exposure to cold stress. The results suggested that the PbbZIP11 gene could participate in cold-stress signaling through the CBF-dependent pathway, which provides a theoretical basis for the PbbZIP11-mediated response to cold stress and for the genetic breeding of pear varieties with low-temperature tolerance.

Sheath blight (ShB) of maize, as a soil-borne disease caused by Rhizoctonia solani AG1-IA, is one of the main obstacles for maintaining the sustainable production of maize. R. solani has a wide host range and low-resistance sources, there is a lack of resistant resources against ShB, and the damage caused by ShB cannot be effectively controlled. To effectively protect crops against ShB, it is necessary to combine information about its pathogenicity and about the disease prevention and control of the pathogenic fungus and to identify areas that require more in-depth research. While progress has been made in the identification of disease-related genes in corn and pathogens, their mechanisms remain unclear. Research related to disease control involves the use of agronomic methods, chemical control, biological control, and genetic improvement. Breeding and identification of high-resistant maize varieties are key and difficult points in the control of maize sheath blight. This article reviews the research progress of the symptoms, the pathogen’s biological characteristics, the infection process, the pathogenic mechanism, and comprehensive control of maize sheath blight disease and provides future research directions of maize sheath blight in China. It aims to provide reliable technical routes and research ideas for future crop-disease-resistance research, especially to sheath blight of maize.

ADP-Glc pyrophosphorylase (AGPase), which catalyzes the transformation of ATP and glucose-1-phosphate (Glc-1-P) into adenosine diphosphate glucose (ADP-Glc), acts as a rate-limiting enzyme in crop starch biosynthesis. Prior research has hinted at the regulation of AGPase by phosphorylation in maize. However, the identification and functional implications of these sites remain to be elucidated. In this study, we identified the phosphorylation site (serine at the 31st position of the linear amino acid sequence) of the AGPase large subunit (Sh2) using iTRAQTM. Subsequently, to ascertain the impact of Sh2 phosphorylation on AGPase, we carried out site-directed mutations creating Sh2-S31A (serine residue replaced with alanine) to mimic dephosphorylation and Sh2-S31D (serine residue replaced with aspartic acid) or Sh2-S31E (serine residue replaced with glutamic acid) to mimic phosphorylation. Preliminary investigations were performed to determine Sh2 subcellular localization, its interaction with Bt2, and the resultant AGPase enzymatic activity. Our findings indicate that phosphorylation exerts no impact on the stability or localization of Sh2. Furthermore, none of these mutations at the S31 site of Sh2 seem to affect its interaction with Bt2 (smaller subunit). Intriguingly, all S31 mutations in Sh2 appear to enhance AGPase activity when co-transfected with Bt2, with Sh2-S31E demonstrating a substantial five-fold increase in AGPase activity compared to Sh2. These novel insights lay a foundational groundwork for targeted improvements in AGPase activity, thus potentially accelerating the production of ADP-Glc (the primary substrate for starch synthesis), promising implications for improved starch biosynthesis, and holding the potential to significantly impact agricultural practices.

Starch phosphorylase (PHO) is a pivotal enzyme within the GT35-glycogen–phosphorylase (GT; glycosyltransferases) superfamily. Despite the ongoing debate surrounding the precise role of PHO1, evidence points to its substantial influence on starch biosynthesis, supported by its gene expression profile and subcellular localization. Key to PHO1 function is the enzymatic regulation via phosphorylation; a myriad of such modification sites has been unveiled in model crops. However, the functional implications of these sites remain to be elucidated. In this study, we utilized site-directed mutagenesis on the phosphorylation sites of Zea mays PHO1, replacing serine residues with alanine, glutamic acid, and aspartic acid, to discern the effects of phosphorylation. Our findings indicate that phosphorylation exerts no impact on the stability or localization of PHO1. Nonetheless, our enzymatic assays unveiled a crucial role for phosphorylation at the S566 residue within the L80 region of the PHO1 structure, suggesting a potential modulation or enhancement of PHO1 activity. These data advance our understanding of starch biosynthesis regulation and present potential targets for crop yield optimization.

The efficacy of government interventions in epidemic has become a hot subject since the onset of COVID-19. There is however much variation in the results quantifying the effects of interventions, which is partly related to the varying modelling approaches employed by existing studies. Among the many factors affecting the modelling results, people’s voluntary behavior change is less examined yet likely to be widespread. This paper therefore aims to analyze how the choice of modelling approach, in particular how voluntary behavior change is accounted for, would affect the intervention effect estimation. We conduct the analysis by experimenting different modelling methods on a same data set composed of the 500 most infected U.S. counties. We compare the most frequently used methods from the two classes of modelling approaches, which are Bayesian hierarchical model from the class of computational approach and difference-in-difference from the class of natural experimental approach. We find that computational methods that do not account for voluntary behavior changes are likely to produce larger estimates of intervention effects as assumed. In contrast, natural experimental methods are more likely to extract the true effect of interventions by ruling out simultaneous behavior change. Among different difference-in-difference estimators, the two-way fixed effect estimator seems to be an efficient one. Our work can inform the methodological choice of future research on this topic, as well as more robust re-interpretation of existing works, to facilitate both future epidemic response plans and the science of public health.

ADP-Glc pyrophosphorylase (AGPase) is a pivotal enzyme catalyzing the conversion of ATP and glucose-1-phosphate (Glc-1-P) to adenosine diphosphate glucose (ADP-Glc), thereby serving as a rate-limiting factor in starch biosynthesis in crops. Although previous investigations have suggested phosphorylation-based regulation of AGPase in maize, the explicit modulation mechanisms have yet to be elucidated. This research evaluated the effect of point mutations at phosphorylation sites (identified using iTRAQTM AB SCIEX, Framingham, MA, USA) on the subcellular localization and activity of the AGPase small subunit Bt2, and its interaction with the large subunit Sh2, in maize. Despite the induction of point mutations, subcellular localization of the Bt2 subunit remained unaltered, primarily within the cytoplasm and nucleus. The interaction between Bt2 and Sh2 subunits continued, mainly in the chloroplast. Notably, an increase in AGPase activity was observed in the case of simulated phosphorylation point mutations, whereas dephosphorylation activity significantly diminished relative to the wild type. These findings demonstrate that point mutations do not affect the subcellular localization of the Bt2 subunit or its interaction with the Sh2 subunit, but substantially modulate AGPase activity. This study provides critical insights into the role of point mutations in enhancing AGPase activity, thus potentially accelerating the production of ADP-Glc, the primary substrate for starch synthesis, promising implications for improved starch biosynthesis in maize.

In order to investigate the crucial role of ZmBT1 in starch accumulation during maize grain development and analyze the expression and distribution of ZmBT1 in various maize tissues, we prepared a polyclonal antibody. Specifically, we successfully expressed the recombinant plasmid pGEX-6p-ZmBT1-C (382-437aa) and purified Gst-ZmBT1-C as the antigen for antibody preparation. Our results confirmed that the ZmBT1 protein in maize tissues can be specifically recognized by the ZmBT1 antibody. Through Western blotting, we observed that the expression protein of ZmBT1 varied by tissues, with the highest content in the grain and endosperm. Furthermore, we employed a combination of Western blotting and quantitative real-time PCR to show that the expression level of ZmBT1 can be influenced by plant hormones. This finding suggests that ZmBT1 plays a critical role in the accumulation of starch and opens up new avenues for functional studies of this protein.

Russeting, a disorder of pear fruit skin, is mainly caused by suberin accumulation on the inner part of the outer epidermal cell layers. ABA was identified as a crucial phytohormone in suberification. Here, we demonstrated that the ABA content in russet pear skin was higher than in green skin. Then, ABA was applied to explore the changes in phenotype and suberin composition coupled with RNA-Seq and metabolomics to investigate the probably regulatory pathway of ABA-mediated suberification. The results showed that ABA treatment increased the expression of ω-3 fatty acid desaturase (FAD) and the content of α-linolenic acid. We identified 17 PbFADs in white pear, and the expression of PbFAD3a was induced by ABA. In addition, the role of PbFAD3a in promoting suberification has been demonstrated by overexpression in Arabidopsis and VIGS assays in the fruitlets. GUS staining indicated that the promoter of PbFAD3a was activated by ABA. Furthermore, MYC2 and MYB1R1 have been shown to bind to the PbFAD3a promoter directly and this was induced by ABA via yeast one-hybrid (Y1H) screening and qRT–PCR. In summary, our study found that ABA induces the expression of MYC2 and MYB1R1 and activates the PbFAD3a promoter, contributing to the formation of russet pear skin. Functional identification of key transcription factors will be the goal of future research. These findings reveal the molecular mechanism of ABA-mediated suberization in the russet skin and provide a good foundation for future studies on the formation of russet skin.

Iron is an essential trace element for plants; however, low bioactive Fe in soil continuously places plants in an Fe-deficient environment, triggering oxidative damage. To cope with this, plants make a series of alterations to increase Fe acquisition; however, this regulatory network needs further investigation. In this study, we found notably decreased indoleacetic acid (IAA) content in chlorotic pear (Pyrus bretschneideri Rehd.) leaves caused by Fe deficiency. Furthermore, IAA treatment slightly induced regreening by increasing chlorophyll synthesis and Fe2+ accumulation. At that point, we identified PbrSAUR72 as a key negative effector output of auxin signaling and established its close relationship to Fe deficiency. Furthermore, the transient PbrSAUR72 overexpression could form regreening spots with increased IAA and Fe2+ content in chlorotic pear leaves, whereas its transient silencing does the opposite in normal pear leaves. In addition, cytoplasm-localized PbrSAUR72 exhibits root expression preferences and displays high homology to AtSAUR40/72. This promotes salt tolerance in plants, indicating a putative role for PbrSAUR72 in abiotic stress responses. Indeed, transgenic plants of Solanum lycopersicum and Arabidopsis thaliana overexpressing PbrSAUR72 displayed less sensitivity to Fe deficiency, accompanied by substantially elevated expression of Fe-induced genes, such as FER/FIT, HA, and bHLH39/100. These result in higher ferric chelate reductase and root pH acidification activities, thereby hastening Fe absorption in transgenic plants under an Fe-deficient condition. Moreover, the ectopic overexpression of PbrSAUR72 inhibited reactive oxygen species production in response to Fe deficiency. These findings contribute to a new understanding of PbrSAURs and its involvement in Fe deficiency, providing new insights for the further study of the regulatory mechanisms underlying the Fe deficiency response.

AIM: To evaluate the influence of pars plana vitrectomy(PPV)on ocular surface using Keratograph 5M. METHODS: Totally 30 consecutive patients(30 eyes)undergoing primary 23G PPV were recruited in the study. Ocular Surface Disease Index(OSDI)questionnaire was performed. Ocular surface parameters, including tear meniscus height(TMH), noninvasive tear break up time(NITBUT)and bulbar redness score were obtained preoperatively, in 2, 4, 8 and 12wk postoperatively by Keratograph 5M. Correlations between all the clinical parameters were analyzed further.RESULTS: The percentages of both photophobia and gritty within 4wk after PPV were significantly higher than preoperation, while they decreased to the preoperative levels in both 8wk and 12wk postoperatively. The percentage of sore eyes in the first week postoperatively was significantly higher than preoperation, but there were no significant differences between the percentages of preoperation and 2, 4, 8 and 12wk postoperatively. OSDI score increased significantly within 4wk postoperatively, but it returned to the preoperative level in 8 and 12wk. TMH increased with 2wk postoperatively, but there were no significant differences compared with preoperation. Both NITBUT-first and NITBUT-average shortened significantly within 8wk postoperatively, but they gradually improved to the preoperative levels in 12wk. Bulbar redness score was significantly higher than the preoperative level within 4wk postoperatively, but it returned to the preoperative level in 8wk. NITBUT-first and NITBUT-average had a significant positive correlation at each visit. TMH had a significant positive correlation with NITBUT-average in the first week postoperatively.CONCLUSION: Keratograph 5M can provide a reliable noninvasive method to assess the influence of PPV on the ocular surface. PPV may cause various changes in both symptoms and signs of ocular surface damages at the early stage, while all these changes will return to preoperative levels gradually in 12wk postoperatively.

Objective The cerebrospinal fluid (CSF) cytological and imaging features in 4 patients with meningeal carcinomatosis (MC) were investigated to assist with the diagnosis and improve the treatment of the disease. Methods A retrospective analysis was utilized to investigate the clinical manifestations, imaging, and liquid⁃based CSF cytology of 4 patients with MC by using CSF cytology, all of whom were treated at the Department of Neurology at Wuhan No.1 Hospital from February 2016 to January 2019. Results Four patients (2 males and 2 females) with an average age of (60.00±15.85) years old had a symptom ofheadache,2 of whom had a symptom of slow response. Among 4 patients, all had a history of tumor disease; 2 had a history of lung cancer; one had a history of gastric cancer; one had a history of breast cancer. Four patients underwent lumbar puncture. Two patients had increased white blood cell count, and the other 2 demonstrated decreased glucose levels. Three patients had elevated protein levels. There were 3 cases with increased intracranial pressure (ICP) and 4 cases with atypical cells, as indicated byliquid⁃based CSF cytology. However, no apparent meningeal enhancement was found in 4 patients. One of the patients had a negative PET⁃CT. All patients had been receiving or received symptomatic treatment during hospitalization. Post hospital discharge, one patient had been in treatment, and the other 3 patients died within one month to one year after diagnosis. Conclusions The imaging findings of patients with MC were atypical. The clinical history of patients with MC can help with diagnosis. The CSF cytology on these patients has considerable significance to diagnosis. DOI：10.3969/j.issn.1672⁃6731.2020.07.015

Vertigo is a common disease, which is a kind of motor or positional illusion caused by the obstacle of spatial positioning. Qianglidingxuan Tablets (QLDX) tablets tablet has a significant effect in the treatment of vertigo, but its active components and mechanism is not clear. In this study, we explored the effective components and mechanism of QLDX tablets in treating vertigo using network pharmacology and molecular docking. Herein, we found 163 potential targets corresponding to 60 bio-active components of QLDX tablets, and 160 known vertigo-related targets. Among which, 41 targets were shared between potential targets of QLDX tablets and known vertigo related targets. The mainly active components included flavonoids, volatile oil, alkaloid, and polysaccharides. Potential targets mainly regulating of cell response to chemical stress, oxidative stress, reactive oxygen species, epithelial cell proliferation, lipopolysaccharide, apoptosis signal pathway and steroid hormones. Additionally, molecular docking verified that the active compounds suchilactone in Gastrodia rhizome, epicatechin and liriodendrin in Eucommia ulmoides bark, myricanone in Ligusticum chuaxion, acacetin-7-O-β-D-galactopyranoside in Wild chrysanthemum, and kaempferol in Eucommia ulmoides leaves with the top core genes ESR, AR, ERBB2, EGFR, MAPK1 and CASP8. This work systematically explored the anti-vertigo mechanism of QLDX tablets and provides a novel perspective for future pharmacological research.

Nimbolide, a triterpenoid isolated from flower of neem tree possess various therapeutic properties. The objective of the study was to assess the anti-arthritic activity of nimbolide in arthritis induced rats. Nimbolide (20 mg/kg per day) was given orally to arthritic rats induced with Complete Freund’s Adjuvant and changes in paw volume, body weight, organ indices (thymus and spleen), arthritic score, biochemical parameters and proinflammatory cytokines levels were determined. Histopathological analysis was also performed. Western blot analysis was also performed. Rats treated with nimbolide displayed marked reduction in arthritic score, organ indices, volume of paw, edema formation, along with substantial enhancement in body weight. Histopathological findings showed significant reduction in destruction of joints and inflammation following nimbolide treatment. The protective action of arthritic rats treated with nimbolide was also substantiated by molecular and biochemical studies. The results of the study show that nimbolide treatment has markedly enhanced health and reduced inflammation via lessening the proinflammatory cytokines expression in arthritic rats. Hence, nimbolide may be used as a potent therapeutic drug in treating rheumatoid arthritis.

Starch biosynthesis is a complex and highly controlled process that requires coordinated activities among multiple enzymes. Starch synthase III (SSIII) is the largest protein in the starch synthase complex and its function is to lengthen long-chain amylopectin in starch synthesis. It potentially affects the activity of other key enzymes in starch synthesis through protein–protein interactions; therefore, its function and regulation play a predominant role in starch synthesis. In this review, we summarized the main research of SSIII including its biochemical characteristics, structural features, expression atlas, and regulation means. Structural features and expressional analysis indicated that SSIIIa is the main functional protein in maize endosperm rather than SSIIIb-a and SSIIIb-b, even though they are similar in the tertiary structures. The regulation investigation of SSIIIa showed that there are 13 transcription factors that control the transcription of SSIIIa. Interaction network analysis showed that SSIIIa could be involved with ten other key enzymes in starch synthesis. In conclusion, this review considerably extends our understanding of SSIII and provides the theoretical basis for improving starch synthesis by SSIII in maize.

With the development of a large number of tall dams and large cascade reservoir projects in the Lantsang River Basin, a large water level fluctuating zone (WLFZ) containing cascading reservoirs has formed. This newborn ecosystem is related to the sustainable development of hydropower projects, and has become a new problem to be studied urgently. Taking WLFZs in the Huangdeng, Xiaowan and Nuozhadu Reservoirs in the Lantsang River Basin as study areas, this study used multi-spectral remote-sensing field data obtained with unmanned aerial vehicles (UAVs) to ascertain the species types, coverage, distribution characteristics, dominant species and pioneer species of naturally restored vegetation. The considered data were subjected to UAV data processing, vegetation classification using multi-spectral images and a geographic information system (GIS) terrain-distribution analysis. Results show that: Polygonum Plebeium, Cynodon dactylon, Xanthium sibiricum, Ageratum conyzoides, Eleusine indica, Digitaria sanguinalis and Verbena officinalis are the dominant species of vegetation that could be naturally restored in the WLFZ; the vegetation coverage and the number of species are significantly positively correlated with the age and restoration periods of the WLFZ; the vegetation coverage of each study area increases at first, and then decreases, as a function of elevation; gentle slopes about 0-25°are more suitable for vegetation restoration. This study provides first-hand data on the natural restoration of vegetation in WLFZs, and gives a useful reference for its ecological restoration as a consequence of hydropower cascade development in the Lantsang River Basin. Finally, the study demonstrates that light UAV remote sensing is an attractive choice for investigating vegetation in reservoir WLFZs.

In this paper, a coupling nonlinear dynamic model of the drum and subgrade is established for the vibratory roller. The dynamic characteristics of the rigid drum of the vibratory roller in the process of vibratory compaction are comprehensively investigated by time history, phase diagram, frequency spectrum, Poincare map, and bifurcation diagram. During the compaction process, the stiffness of the subgrade increases and the motion of the rigid drum of the vibratory roller changes from a single period to multiple periods and finally enters chaos by the way of period doubling. Moreover, the roller parameters also significantly affect the dynamic characteristics of the rigid drum and the compaction effect of the subgrade. Based on detailed numerical results, a parameter adjustment strategy about the roller frequency and nominal amplitude is proposed, which can avoid the “bouncing” of the drum during compaction and improve the compaction efficiency.

Starch phosphorylase (PHO) is a multimeric enzyme with two distinct isoforms: plastidial starch phosphorylase (PHO1) and cytosolic starch phosphorylase (PHO2). PHO1 specifically resides in the plastid, while PHO2 is found in the cytosol. Both play a critical role in the synthesis and degradation of starch. This study aimed to report the detailed structure, function, and evolution of genes encoding PHO1 and PHO2 and their protein ligand-binding sites in eight monocots and four dicots. “True” orthologs of PHO1 and PHO2 of Oryza sativa were identified, and the structure of the enzyme at the protein level was studied. The genes controlling PHO2 were found to be more conserved than those controlling PHO1; the variations were mainly due to the variable sequence and length of introns. Cis-regulatory elements in the promoter region of both genes were identified, and the expression pattern was analyzed. The real-time quantitative polymerase chain reaction indicated that PHO2 was expressed in all tissues with a uniform pattern of transcripts, and the expression pattern of PHO1 indicates that it probably contributes to the starch biosynthesis during seed development in Zea mays. Under abscisic acid (ABA) treatment, PHO1 was found to be downregulated in Arabidopsis and Hordeum vulgare. However, we found that ABA could up-regulate the expression of both PHO1 and PHO2 within 12 h in Zea mays. In all monocots and dicots, the 3D structures were highly similar, and the ligand-binding sites were common yet fluctuating in the position of aa residues.

A framework is established for evaluating the adaptability of rural households to tourism disturbance in the suburbs of a metropolis, based on the theory of the social-ecological system with tourism as the system disturbance. Two traditional villages in Beijing suburbs are selected as study cases and compared in terms of the development stage of tourism and the phases and approaches of adaption by rural households to tourism disturbance. Stepwise regression and grey relational analysis are applied to investigate the factors influencing the functionality and stability of the rural social-ecological system.

Starch phosphorylase is a member of the GT35-glycogen-phosphorylase superfamily. Glycogen phosphorylases have been researched in animals thoroughly when compared to plants. Genetic evidence signifies the integral role of plastidial starch phosphorylase (PHO1) in starch biosynthesis in model plants. The counterpart of PHO1 is PHO2, which specifically resides in cytosol and is reported to lack L80 peptide in the middle region of proteins as seen in animal and maltodextrin forms of phosphorylases. The function of this extra peptide varies among species and ranges from the substrate of proteasomes to modulate the degradation of PHO1 in Solanum tuberosum to a non-significant effect on biochemical activity in Oryza sativa and Hordeum vulgare. Various regulatory functions, e.g., phosphorylation, protein–protein interactions, and redox modulation, have been reported to affect the starch phosphorylase functions in higher plants. This review outlines the current findings on the regulation of starch phosphorylase genes and proteins with their possible role in the starch biosynthesis pathway. We highlight the gaps in present studies and elaborate on the molecular mechanisms of phosphorylase in starch metabolism. Moreover, we explore the possible role of PHO1 in crop improvement.

Nonlinear dynamic modeling and model reduction strategy are studied in this paper for a rotating thin cylindrical shell. The nonlinear dynamic model is first established in terms of ordinary differential equations, in which the effects of Coriolis and centrifugal forces are considered, as well as the initial hoop tension due to rotation. This model describes both the in-plane vibrations and the flexural vibration and reflects the coupling effects of those deformations. Based on this original model, a novel model reduction strategy is proposed to reduce the degrees of freedom by neglecting vibration modes predominated by in-plane vibrations. Meanwhile, for the reduced-order model, the in-plane vibrations’ contributions to the rotating shell’s response are still preserved. To validate the dynamic model and the model reduction strategy, comparisons and simulations are carried out. Subsequently, nonlinear dynamic behaviors are investigated preliminarily by analyzing the rotating cylindrical shell’s amplitude-frequency responses under different excitation levels.

Alligator weed is reported to have a strong ability to adapt to potassium deficiency (LK) stress. Leaves are the primary organs responsible for photosynthesis of plants. However, quantitative proteomic changes in alligator weed leaves in response to LK stress are largely unknown. In this study, we investigated the physiological and proteomic changes in leaves of alligator weed under LK stress. We found that chloroplast and mesophyll cell contents in palisade tissue increased, and that the total chlorophyll content, superoxide dismutase (SOD) activity and net photosynthetic rate (PN) increased after 15 day of LK treatment, but the soluble protein content decreased. Quantitative proteomic analysis suggested that a total of 119 proteins were differentially abundant proteins (DAPs). KEGG analysis suggested that most represented DAPs were associated with secondary metabolism, the stress response, photosynthesis, protein synthesis, and degradation pathway. The proteomic results were verified using parallel reaction monitoring mass spectrometry (PRM–MS) analysis and quantitative real-time PCR (qRT-PCR)assays. Additional research suggested that overexpression of cationic peroxidase 1 of alligator weed (ApCPX1) in tobacco increased LK tolerance. The seed germination rate, peroxidase (POD) activity, and K+ content increased, and the hydrogen peroxide (H2O2) content decreased in the three transgenic tobacco lines after LK stress. The number of root hairs of the transgenic line was significantly higher than that of WT, and net K efflux rates were severely decreased in the transgenic line under LK stress. These results confirmed that ApCPX1 played positive roles in low-K+ signal sensing. These results provide valuable information on the adaptive mechanisms in leaves of alligator weed under LK stress and will help identify vital functional genes to apply to the molecular breeding of LK-tolerant plants in the future.

Purpose. To analyze foveal microvascular abnormalities in different stages of diabetic retinopathy (DR) using optical coherence tomography angiography (OCTA) with projection artifact removal (PAR). Methods. We analyzed 93 eyes of 59 patients with diabetes—31 with no DR (no DR), 34 with mild to moderate nonproliferative DR (mild DR), and 28 with severe nonproliferative DR to proliferative DR (severe DR)—and 31 age-matched healthy controls. Sections measuring 3 × 3 mm2 centered on the fovea were obtained using OCTA. The area, perimeter, and acircularity index (AI) of the foveal avascular zone (FAZ), vessel density within a 300 μm wide region of the FAZ (FD-300), and parafoveal vessel density in the superficial capillary plexus (SCP) and deep capillary plexus (DCP) were calculated using novel built-in software with PAR. Results. There was no statistically significant difference in the FAZ area (p=0.162). There was a statistically significant difference in the FAZ perimeter (p=0.010) and the AI (p

The purpose of this study was to explore the midterm clinical outcomes of unicompartmental knee replacement (UKR) for medial knee arthropathy through a minimally invasive approach (MIA).From January 2006 to June 2010, 442 consecutive patients (485 knees) were included in the study. All patients underwent MIA-UKR with the mobile bearing Oxford phrase III prosthesis. The incision was made starting 1 cm medial to the medial pole of the patella and extending distally to the tibial tubercle. Radiographic evaluations include femorotibial angle (FTA) from coronal x-rays and rectified varus deformity angle, while clinical evaluations included Knee Society Score (KSS, clinical score and function score), the Western Ontario and McMaster Universities Arthritis Index (WOMAC) osteoarthritis index and visual analog scale (VAS) for pain. Patients followed-up at 1, 3, 6, 12 months after surgery and each year thereafter.Four hundreds and two patients completed the entire follow-up, 40 patients (45 knees) were lost to follow-up. The average follow-up time was 73.0 ± 1.9 months. The mean length of the incisions was 5.0 ± 0.2 cm. The average FTA decreased from 183.6° ± 5.1° preoperatively to 174.3° ± 4.2° postoperatively, and the mean rectified varus deformity angle was 9.3° ± 1.2°. The KSS clinical score improved from 42.4 ± 2.9 to 92.9 ± 3.8, and the function score improved from 53.5 ± 3.8 to 93.5 ± 4.0. The WOMAC score improved from 47.5 ± 3.1 preoperatively to 12.3 ± 1.5 at the last evaluation. The VAS dropped from 7.8 ± 1.9 preoperatively to 1.6 ± 0.2 postoperatively. All clinical evaluations (KSS, WOMAC, VAS) were significantly different (p < 0.05) from pre and post-operative evaluations. The survival rate was 99.1% at 73 months, and the revision rate was 0.9%.The midterm clinical outcomes of MIA-UKR are satisfactory in a Chinese patient population, which is a good surgical option for patients with medial arthropathy of the knee. However, longer-term follow-up studies should be performed in these patients.

The present study aimed to determine the optimal posterior tibial plateau inclination for fixed-platform unicondylar knee arthroplasty (UKA) using finite element analysis (FEA). These findings provided a theoretical basis for selecting an appropriate posterior inclination of the tibial plateau during surgery. The present study utilized the FEA method to create models of fixed-platform UKA with tibial plateau posterior inclinations of 3, 6 and 9˚. The stress changes in the internal structures of each model after knee flexion motion were then compared. During knee flexion from 0 to 90˚, the contact and Von Mises equivalent stresses of the femoral condyle prosthesis and tibial platform pad revealed consistent trends of 3˚ posterior inclination, >6˚ posterior inclination and >9˚ posterior inclination. The present study established the first quasi-dynamic fixed-platform UKA model of the knee joint under load-bearing conditions. From a theoretical perspective, it was found that controlling the posterior inclination of UKA between 6 and 9˚ may be more beneficial for the survival of the tibial platform pad than between 3 and 6˚. It is also more effective in reducing pad wear. [ABSTRACT FROM AUTHOR]