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Background: Postoperative liver metastasis significantly impacts the prognosis of pancreatic neuroendocrine tumor (panNET) patients after R0 resection. Combining computational pathology and deep learning radiomics can enhance the detection of postoperative liver metastasis in panNET patients., Methods: Clinical data, pathology slides, and radiographic images were collected from 163 panNET patients post-R0 resection at Fudan University Shanghai Cancer Center (FUSCC) and FUSCC Pathology Consultation Center. Digital image analysis and deep learning identified liver metastasis-related features in Ki67-stained whole slide images (WSIs) and enhanced CT scans to create a nomogram. The model's performance was validated in both internal and external test cohorts., Results: Multivariate logistic regression identified nerve infiltration as an independent risk factor for liver metastasis (p < 0.05). The Pathomics score, which was based on a hotspot and the heterogeneous distribution of Ki67 staining, showed improved predictive accuracy for liver metastasis (AUC = 0.799). The deep learning-radiomics (DLR) score achieved an AUC of 0.875. The integrated nomogram, which combines clinical, pathological, and imaging features, demonstrated outstanding performance, with an AUC of 0.985 in the training cohort and 0.961 in the validation cohort. High-risk group had a median recurrence-free survival of 28.5 months compared to 34.7 months for the low-risk group, showing significant correlation with prognosis (p < 0.05)., Conclusion: A new predictive model that integrates computational pathologic scores and deep learning-radiomics can better predict postoperative liver metastasis in panNET patients, aiding clinicians in developing personalized treatments., (© 2024. The Author(s).)

The Tibetan antelope (Pantholops hodgsonii), blue sheep (Pseudois nayaur), and Tibetan sheep (Ovis aries) are the dominant small ruminants in the Three-River-Source National Park (TRSNP). However, knowledge about the association between gut microbiota and host adaptability remains poorly understood. Herein, multi-omics sequencing approaches were employed to investigate the gut microbiota-mediated forage adaption in these ruminants. The results revealed that although wild ruminants (WR) of P. hodgsoni and P. nayaur were faced with severe foraging environments with significantly low vegetation coverage and nutrition, the apparent forage digestibility of dry matter, crude protein, and acid detergent fiber was significantly higher than that of O. aries. The 16s rRNA sequencing showed that the gut microbiota in WR underwent convergent evolution, and alpha diversity in these two groups was significantly higher than that in O. aries. Moreover, indicator species, including Bacteroidetes and Firmicutes, exhibited positive relationships with apparent forage digestibility, and their relative abundances were enriched in the gut of WR. Enterotype analysis further revealed that enterotype 1 belonged to WR, and the abundance of fatty acid synthesis metabolic pathway-related enzyme genes was significantly higher than enterotype 2, represented by O. aries. Besides, the metagenomic analysis identified 14 pathogenic bacterial species, among which 10 potentially pathogenic bacteria were significantly enriched in the gut microbiota of O. aries. Furthermore, the cellulolytic strains and genes encoding cellulase and hemicellulase were significantly enriched in WR. In conclusion, our results provide new evidence of gut microbiota to facilitate wildlife adaption in severe foraging environments of the TRSNP, China., (© 2024 The Authors. Integrative Zoology published by International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

Objective: To investigate the time course of action of different doses of mivacurium and determine the appropriate dose for laryngeal mask airway (LMA) insertion for day-case urologic surgery in children., Methods: A total of 105 patients who enrolled in this study between March 2021 and December 2021 were randomised into 3 groups: Group A (mivacurium 0.15 mg/kg, n  = 35), Group B (mivacurium 0.20 mg/kg, n  = 35) and Group C (mivacurium 0.25 mg/kg, n  = 35). The different doses of mivacurium were injected before LMA insertion. The primary outcomes included the grading of conditions for the LMA insertion-18 score, onset time, recovery index and the duration that mivacurium was effective. Secondary outcomes included pulse oxygen saturation, mean blood pressure, heart rate and the incidence of adverse events., Results: The score of the conditions for LMA insertion in Group A was significantly lower than in Groups C and B ( p  < 0.005). There was a significant difference in the onset time between Groups B and A ( p  < 0.005). There was no significant difference in the overall incidence of adverse reactions between these groups ( p  > 0.05)., Conclusion: Anaesthesia with 0.2 mg/kg of mivacurium can effectively shorten the onset time and facilitate insertion of the LMA in children undergoing day-case urologic surgery., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Ye, Nian, Zhou, Xie, Li, Xue and Han.)

Phospholipase C epsilon 1 (PLCE1) is a well-established susceptibility gene for esophageal squamous cell carcinoma (ESCC). Identification of the underlying mechanism(s) regulated by PLCE1 could lead to a better understanding of ESCC tumorigenesis. In this study, we found that PLCE1 enhances tumor progression by regulating the replicative helicase MCM7 via two pathways. PLCE1 activated PKCα-mediated phosphorylation of E2F1, which led to the transcriptional activation of MCM7 and miR-106b-5p. The increased expression of miR-106b-5p, located in intron 13 of MCM7, suppressed autophagy and apoptosis by targeting Beclin-1 and RBL2, respectively. Moreover, MCM7 cooperated with the miR-106b-25 cluster to promote PLCE1-dependent cell-cycle progression both in vivo and in vitro. In addition, PLCE1 potentiated the phosphorylation of MCM7 at six threonine residues by the atypical kinase RIOK2, which promoted MCM complex assembly, chromatin loading, and cell-cycle progression. Inhibition of PLCE1 or RIOK2 hampered MCM7-mediated DNA replication, resulting in G1-S arrest. Furthermore, MCM7 overexpression in ESCC correlated with poor patient survival. Overall, these findings provide insights into the role of PLCE1 as an oncogenic regulator, a promising prognostic biomarker, and a potential therapeutic target in ESCC., Significance: PLCE1 promotes tumor progression in ESCC by activating PKCα-mediated phosphorylation of E2F1 to upregulate MCM7 and miR-106b-5p expression and by potentiating MCM7 phosphorylation by RIOK2., (©2023 American Association for Cancer Research.)

The experiment was conducted at the Taidong Base of Shandong Institute of Pomology, Tai'an City, Shandong Province, China, from May to October 2022. Using Fuji Apple Tianhong.2/SH/ Malus robusta ( M. robusta ) as experimental materials, the differences and mechanisms of the effects of non-bagging and bagging cultivation on the growth and changes in some substance content of Fuji apple fruits were studied. The results showed that compared with bagging, non-bagging cultivation increased single fruit weight and decreased fruit shape index, increased fruit sugar content, reduced acid content, and increased taste. It increased the content of vitamin C (VC) and protein in the fruit, increased the types and content of aromatic components in the fruit, significantly increased the activity of sugar- and acid-related enzymes, and improved the antioxidant capacity of the fruit. Compared to bagging cultivation, non-bagging cultivation improved the weight, taste (sugar acid ratio), and aroma of Fuji fruit, which is related to increasing the content of auxin (IAA), cytokinin (ZR), and salicylic acid (SA) and reducing the content of abscisic acid (ABA) in the fruit, as well as increasing the content of medium and trace elements calcium (Ca), iron (Fe), manganese (Mg), and boron (B). One of the mechanisms involved is the significant increase in gene expression related to phenylpropanoid biosynthesis, pentose and glucuronate interconversion, starch and sucrose metabolism, zeatin biosynthesis, microtubules, motor proteins microtubule movement, xyloglucan metabolic process, cell division, and peroxidase activity. In short, non-bagging cultivation is more conducive to improving the intrinsic quality and flavor development of Fuji apples, and one of the mechanisms is that non-bagging cultivation is beneficial for increasing the expression of related genes.

Introduction: Apricot fruits are edible and serve as a source of medicinal compounds. Flavonols are important plant secondary metabolites that have antioxidant and antitumor effects and may promote cardiovascular health., Methods: The flavonoid content in three stages of the 'Kuijin' and the 'Katy' was observed, followed by the combination of metabolome and transcriptome analysis to explore the metabolic basis of flavonol synthesis., Results: The differences in the metabolite contents between stages (of the same cultivar) and between cultivars (at the same stage) revealed decreases in the flavonoid content as fruits developed (i.e., from 0.28 mg/g to 0.12 mg/g in 'Kuijin' and from 0.23 mg/g to 0.05 mg/g in 'Katy'). To decipher the regulation of flavonol synthesis in apricot (Prunus armeniaca L.), the metabolomes and transcriptomes of fruit pulp at three developmental stages of 'Kuijin' and the 'Katy' were analyzed. A total of 572 metabolites were detected in 'Kuijin' and the 'Katy' pulp, including 111 flavonoids. The higher flavonol content young 'Kuijin' fruits at 42 days after full bloom is mainly due to 10 types of flavonols. Three pairs of significant differences in flavonol content were identified. From these three comparison groups, three structural genes were strongly correlated with the levels of 10 types of flavonols (Pearson correlation coefficients > 0.8, p value < 0.05), including PARG09190, PARG15135, and PARG17939. The weighted gene co-expression network analysis showed that the turquoise module genes were highly correlated with flavonol contents (P < 0.01). There were 4897 genes in this module. Out of 4897 genes, 28 transcription factors are associated with 3 structural genes based on weight value. Two of the transcription factors are not only associated with PARG09190 but also with PARG15135, indicating their critical importance in the flavonols biosynthesis. The two TFs are PARG27864 and PARG10875., Discussion: These findings provide new insights into the biosynthesis of flavonols and may explain the significant differences in flavonoid content between the 'Kuijin' and the 'Katy' cultivars. Moreover, it will aid in genetic improvement to enhance the nutritional and health value of apricots., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Han, Wang, Wang, Dong, Nie and Xue.)

Backgrounds: Transforming growth factor-β (TGF-β) type I receptor (TGFβRI) and type II receptor (TGFβRII) are the members of the TGFβ superfamily, which are potent regulators of cell proliferation and differentiation in many organ systems, and they play key roles in multiple aspects of follicle development., Objectives: We aimed to explore the characterization, expression analysis of TGFβRI and TGFβRII genes, and the association with litter size in Tibetan sheep., Methods: In this study, we cloned the complete coding sequences of TGFβRI and TGFβRII genes in Tibetan sheep and analyzed their genomic structures., Results: The results showed that percentages of sequences homology of the two proteins in Tibetan sheep were the most similar to Ovis aries (100%), followed by Bos mutus (99%). The RT-qPCR showed that two genes were expressed widely in the different tissues of Tibetan sheep. The TGFβRI expression was the highest in the lung (p < 0.05), followed by the spleen and ovary (p < 0.05). The TGFβRII expression was significantly higher in uterus than that in lung and ovary (p < 0.05). In addition, the χ 2 test indicated that all ewes in the population were in Hardy-Weinberg equilibrium, and the population was in medium or low polymorphic information content status. We also found four Single Nucleotide Polymorphism (SNPs), g.9414A > G, g.28881A > G, g.28809T > C, g.10429G > A in sheep TGFβRI gene and g.63940C > T, g.63976C > T, g.64538C > T, g.64504T > A in TGFβRII gene. Three genotypes, except for g.64504T > A, and three haplotypes were identified in each gene. linkage disequilibrium analysis indicated that there was strong linkage disequilibrium in each gene. The association analysis showed that the four SNPs of TGFβRI were associated with litter size (p < 0.05), and g.63940C > T of TGFβRII was confirmed to be associated with litter size (p < 0.05)., Conclusions: Based on these preliminary results, we can assume that TGFβ receptors (TGFβRI and TGFβRII) may play an important role in sheep reproduction., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Copy number variation (CNV), an important source of genomic structural variation, can disturb genetic structure, dosage, regulation and expression, and is associated with phenotypic diversity and adaptation to local environments in mammals. In the present study, 24 resequencing datasets were used to characterize CNVs in three ecotypic populations of Tibetan sheep and assess CNVs related to domestication and adaptation in Qinghai-Tibetan Plateau. A total of 87,832 CNV events accounting for 0.3% of the sheep genome were detected. After merging the overlapping CNVs, 2777 CNV regions (CNVRs) were obtained, among which 1098 CNVRs were shared by the three populations. The average length of these CNVRs was more than 3 kb, and duplication events were more frequent than deletions. Functional analysis showed that the shared CNVRs were significantly enriched in 56 GO terms and 18 KEGG pathways that were mainly concerned with ABC transporters, olfactory transduction and oxygen transport. Moreover, 188 CNVRs overlapped with 97 quantitative trait loci (QTLs), such as growth and carcass QTLs, immunoglobulin QTLs, milk yield QTLs and fecal egg counts QTLs. PCDH15 , APP and GRID2 overlapped with body weight QTLs. Furthermore, Vst analysis showed that RUNX1 , LOC101104348, LOC105604082 and PAG11 were highly divergent between Highland-type Tibetan Sheep (HTS) and Valley-type Tibetan sheep (VTS), and RUNX1 and LOC101111988 were significantly differentiated between VTS and Oura-type Tibetan sheep (OTS). The duplication of RUNX1 may facilitate the hypoxia adaptation of OTS and HTS in Qinghai-Tibetan Plateau, which deserves further research in detail. In conclusion, for the first time, we represented the genome-wide distribution characteristics of CNVs in Tibetan sheep by resequencing, and provided a valuable genetic variation resource, which will facilitate the elucidation of the genetic basis underlying the distinct phenotypic traits and local adaptation of Tibetan sheep., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hu, Zhang, Li, Liu, Xu, Zhao, Han, Xu, Zhao and Zhang.)

Many studies have demonstrated that anthocyanin synthesis in apple peel is induced by light, but the color of bagged apple peel continues to change under dark conditions after light induction has not been characterized. Here, transcriptional and metabolic changes associated with changes in apple peel coloration in the dark after different light induction treatments were studied. Apple pericarp can achieve a normal color under complete darkness followed by light induction. Metabolomics analysis indicated that the expression levels of cyanidin-3-O-galactoside and cyanidin-3-O-glucoside were high, which might be associated with the red color development of apple peel. Transcriptome analysis revealed high expression levels of MdUFGT s, MdMYB s, and MdNAC s, which might play a key role in light-induced anthocyanin accumulation under dark conditions. 13 key genes related to dark coloring after light induction was screened. The results of this study provide new insights into the mechanism of anthocyanin synthesis under dark conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xue, Tian, Chen, Han, Wang and Zhao.)

It was acknowledged long ago that microorganisms have played critical roles in animal evolution. Tibetan wild asses (TWA, Equus kiang ) are the only wild perissodactyls on the Qinghai-Tibet Plateau (QTP) and the first national protected animals; however, knowledge about the relationships between their gut microbiota and the host's adaptability remains poorly understood. Herein, 16S rRNA and meta-genomic sequencing approaches were employed to investigate the gut microbiota-host associations in TWA and were compared against those of the co-resident livestock of yak ( Bos grunnies ) and Tibetan sheep ( Ovis aries ). Results revealed that the gut microbiota of yak and Tibetan sheep underwent convergent evolution. By contrast, the intestinal microflora of TWA diverged in a direction enabling the host to subsist on sparse and low-quality forage. Meanwhile, high microbial diversity (Shannon and Chao1 indices), cellulolytic activity, and abundant indicator species such as Spirochaetes, Bacteroidetes, Prevotella&#95;1 , and Treponema&#95;2 supported forage digestion and short-chain fatty acid production in the gut of TWA. Meanwhile, the enterotype identification analysis showed that TWA shifted their enterotype in response to low-quality forage for a better utilization of forage nitrogen and short-chain fatty acid production. Metagenomic analysis revealed that plant biomass degrading microbial consortia, genes, and enzymes like the cellulolytic strains ( Prevotella ruminicola, Ruminococcus flavefaciens, Ruminococcus albus, Butyrivibrio fibrisolvens , and Ruminobacter amylophilus ), as well as carbohydrate metabolism genes (GH43, GH3, GH31, GH5, and GH10) and enzymes (β-glucosidase, xylanase, and β-xylosidase, etc.) had a significantly higher enrichment in TWA. Our results indicate that gut microbiota can improve the adaptability of TWA through plant biomass degradation and energy maintenance by the functions of gut microbiota in the face of nutritional deficiencies and also provide a strong rationale for understanding the roles of gut microbiota in the adaptation of QTP wildlife when facing harsh feeding environments., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The Reviewer XZ declared a shared affiliation with the authors HL, NZ, LH, XW, CL, XZ, and SX at the time of the review., (Copyright © 2022 Liu, Han, Zhao, Hu, Wang, Luo, Chen, Zhao and Xu.)

There is an increasing awareness of the adverse environmental effects of the intensive practices used in modern crop farming, such as those that cause greenhouse gas emissions and nutrient leaching. Harnessing beneficial microbes by changing planting practices presents a promising strategy for optimizing plant growth and agricultural sustainability. However, the characteristics of soil microorganisms under different planting patterns remain uncertain. We conducted a study of soil bacterial structure and function under monoculture vs. polyculture planting regimes using 16S rRNA gene sequencing on the Qinghai-Tibet Plateau. We observed substantial variations in bacterial richness, diversity, and relative abundances of taxa between gramineous and leguminous monocultures, as well as between gramineae-legume polycultures. The number of operational taxonomic units and alpha and beta diversity were markedly higher in the leguminous monocultures than in the gramineous monocultures; conversely, network analysis revealed that the interactions among the bacterial genera in the gramineous monocultures were more complex than those in the other two planting regimes. Moreover, nitrogen fixation, soil detoxification, and productivity were increased under the gramineous monocultures; more importantly, low soil-borne diseases (e.g., animals parasitic or symbiont) also facilitated strongly suppressive effects toward soil-borne pathogens. Nevertheless, the gramineae-legume polycultures were prone to nitrate seepage contamination, and leguminous monocultures exhibited strong denitrification effects. These results revealed that the gramineous monoculture is a more promising cropping pattern on the Qinghai-Tibetan Plateau. Understanding the bacterial distribution patterns and interactions of crop-sensitive microbes presents a basis for developing microbial management strategies for smart farming., (Copyright © 2022. Published by Elsevier GmbH.)

Actin cytoskeleton and transcription factors play key roles in plant response to salt stress; however, little is known about the link between the two regulators in response to salt stress. Actin-depolymerizing factors (ADFs) are conserved actin-binding proteins in eukaryotes. Here, we revealed that the expression level of ADF1 was induced by salt stress. The adf1 mutants showed significantly reduced survival rate, increased percentage of actin cable and reduced density of actin filaments, while ADF1 overexpression seedlings displayed the opposite results when compared with WT under the same condition. Furthermore, biochemical assays revealed that MYB73, a R2R3 MYB transcription factor, binds to the promoter of ADF1 and represses its expression via the MYB-binding site core motif ACCTAC. Taken together, our results indicate that ADF1 participates in salt stress by regulating actin organization and may also serve as a potential downstream target of MYB73, which is a negative regulator of salt stress., (© The Author(s) 2021. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In the current research, samples of yak gastrointestinal tracts (GITs) were used to profile the bacterial compositional characteristics using high through-put sequencing technology of 16S RNA amplicon. A total of 6959 OTUs was obtained from 20,799,614 effective tags, among which 751 OTUs were shared by ten sections. A total of 16 known phyla were obtained in all samples-the most abundant phyla were Firmicutes (34.58%), Bacteroidetes (33.96%) and Verrucomicrobia (11.70%). At the genus level, a total of 66 genera were obtained- Rikenellaceae&#95;RC9&#95;gut&#95;group (7.24%), Akkermansia (6.32%) and Ruminococcaceae &#95;UCG-005 (6.14%) were the most abundant. Species of Observed (Sob), Shannon and Chao values of the Stomach were the greatest, followed by the large intestine, while small intestine had the lowest diversity ( p < 0.05). Bacteroidete were more abundant in sections from rumen to duodenum; while Firmicutes were the most abundant in sections from jejunum. ABC transporters (7.82%), Aminoacyl-tRNA biosynthesis (4.85%) and Purine metabolism (3.77%) were the most abundant level-3 pathways in all samples. The results of associated correlation analysis indicated that rectum samples might be used as an estimator of rumen bacterial communities and fermentation. The results of this research enrich the current knowledge about the unique animals of the QTP and extend our insight into GITs microecology of various animals.

The rumen microbiota is strongly associated with host health, nutrient absorption, and adaptability. However, the composition, functioning and adaptability of rumen microbiota in Tibetan sheep (TS) across different phenological periods are unclear. In this study we used sequencing of the V4-V5 region of 16S rRNA, qPCR technology and metagenomics to investigate the adaption of rumen microbiota to forage in different stages of phenology. In a grassy period, due to the high nutritional quality of the forage, TS can produce high concentrations of NH 3 -N and short fatty acids by increasing the content of key bacteria in the rumen, such as Bacteroidetes, Prevotella , Succiniclasticum , Treponema , Butyrivibrio fibrisolvens , Fibrobacter succinogenes , Prevotella ruminicola , Ruminococcus albus , and Ruminococcus flavefaciens to aid in growth. In the withering period, there was a positive correlation between microorganisms which indicated the closely cooperation between microorganisms, and metagenomic analysis showed that the high genes (GHs and CBMs) and subtribe (GH8, GH12, GH45, GH6, GH9, GH5, GH10, GH3, GH52, GH11, GH57, CBM1, CBM4, CBM6, CBM16, CBM37, CBM13, CBM35, CBM42, CBM32, and CBM62) that encode cellulolytic enzymes were significantly increased when the host faced low quantity and quality of forage. Genes involved in metabolic pathways, fatty acid biosynthesis and biosynthesis of antibiotics were significantly enriched, which indicated that rumen microbiota could improve plant biomass deconstruction and energy maintenance in the face of nutritional deficiencies. In the regreen period, both the composition and function of rumen microbiota had obvious disadvantages, therefore, to improve the competitiveness of microorganisms, we suggest TS should be supplemented with high-protein feed. This study is of great significance for exploring the high altitude adaptability of TS., (Copyright © 2020 Liu, Hu, Han, Zhao, Xu, Ma, Wang, Zhang, Kang, Zhao and Xu.)

Tibetan wild asses ( Equus Kiang ) are the only wild species of perissodactyls on the Qinghai-Tibet Plateau and appears on the International Union for Conversation of Nature (IUCN) 2012 Red List of threatened species. Therefore, understanding the gut microbiota composition and function of wild asses can provide a theoretical for the situ conservation of wild animals in the future.In this study, we measured the dry matter digestion by the 4 molar hydrochloric acid (4N HCL) acid-insoluble ash method and analyzed the intestinal microbiota of wild asses and domestic donkeys by high-throughput sequencing of the 16s rDNA genes in V3-V4 regions. The results showed that the dry matter digestion in wild asses was significantly higher than in domestic donkeys ( P  < 0.05). No significant difference in alpha diversity was detected between these two groups. Beta diversity showed that the bacterial community structure of wild asses was acutely different from domestic donkeys. At the phylum level, the two dominant phyla Bacteroidetes and Firmicutes in wild asses were significantly higher than that in domestic donkeys. At the genus level, Ruminococcaceae&#95;NK4A214 , Phascolarctobacterium , Coprostanoligenes&#95;group , Lachnospiraceae&#95;XPB1014&#95;group and Akkermansia in wild asses were significantly higher than in domestic donkeys. Moreover, statistical comparisons showed that 40 different metabolic pathways exhibited significant differences. Among them, 29 pathways had richer concentrations in wild asses than domestic donkeys, mainly included amino acid metabolism, carbohydrate metabolism, and energy metabolism. Of note, network analysis showed that wild asses harbored a relatively more complex bacterial network than domestic donkeys, possibly reflecting the specific niche adaption of gut bacterial communities through species interactions. The overall results indicated that wild asses have advantages over domestic donkeys in dry matter digestion, gut microbial community composition and function, and wild asses have their unique intestinal flora to adapt high altitudes on the Qinghai-Tibet plateau., Competing Interests: The authors declare there are no competing interests., (©2020 Liu et al.)

Esophageal squamous cell carcinoma (ESCC) is one of the deadliest malignant diseases. Multiple studies with large clinic-based cohorts have revealed that variations of phospholipase C epsilon 1 (PLCE1) correlate with esophageal cancer susceptibility. However, the causative role of PLCE1 in ESCC has remained elusive. Here, we observed that hypomethylation-mediated upregulation of PLCE1 expression was implicated in esophageal carcinogenesis and poor prognosis in ESCC cohorts. PLCE1 inhibited cell autophagy and suppressed the protein expression of p53 and various p53-targeted genes in ESCC. Moreover, PLCE1 decreased the half-life of p53 and promoted p53 ubiquitination, whereas it increased the half-life of mouse double minute 2 homolog (MDM2) and inhibited its ubiquitination, leading to MDM2 stabilization. Mechanistically, the function of PLCE1 correlated with its direct binding to both p53 and MDM2, which promoted MDM2-dependent ubiquitination of p53 and subsequent degradation in vitro . Consequently, knockdown of PLCE1 combined with transfection of a recombinant adenoviral vector encoding wild-type p53 resulted in significantly increased levels of autophagy and apoptosis of esophageal cancer in vivo . Clinically, the upregulation of PLCE1 and mutant p53 protein predicted poor overall survival of patients with ESCC, and PLCE1 was positively correlated with p53 in ESCC cohorts. Collectively, this work identified an essential role for PLCE1- and MDM2-mediated ubiquitination and degradation of p53 in inhibiting ESCC autophagy and indicates that targeting the PLCE1-MDM2-p53 axis may provide a novel therapeutic approach for ESCC. SIGNIFICANCE: These findings identify hypomethylation-mediated activation of PLCE1 as a potential oncogene that blocks cellular autophagy of esophageal carcinoma by facilitating the MDM2-dependent ubiquitination of p53 and subsequent degradation. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/11/2175/F1.large.jpg., (©2020 American Association for Cancer Research.)

Deregulation of SOX9 in esophageal cancer has been reported. However, the regulatory mechanisms underlying SOX9 during esophageal squamous cell carcinoma (ESCC) progression remain poorly understood. Here, we independently confirmed the increased SOX9 expression in two ESCC cohorts and its correlation with poor prognosis. We demonstrated that SOX9 was required for maintaining self-renewal, motility, and chemoresistance in vitro and that ectopic expression of SOX9 promoted tumorigenicity in vivo. Screening for potential SOX9-regulated miRNAs revealed that target genes of differentially expressed miRNAs were enriched in the PI3K/AKT signaling pathway and identified the downregulated miR-203a as a candidate. Mechanistically, SOX9 activation caused repression of miR-203a transcription by binding to miR-203a promoter, thus preventing the miR-203a-mediated inhibition of multiple PI3K/AKT/mTOR components, including PIK3CA, AKT2, and RPS6KB1. The association between SOX9 expression and PI3K/AKT/mTOR signaling was further validated in clinical samples. Moreover, rapamycin treatment attenuated the SOX9-mediated malignant phenotypes and potentiated cisplatin-mediated inhibition of tumor growth. Together, these findings uncover a novel activation of the PI3K/AKT pathway by the SOX9/miR-203a axis and define a subgroup of patients who may benefit from targeted therapy., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The rumen microbiota of ruminants plays a vital role in fiber digestion, and environmental factors affect its community structure. The yak ( Bos grunniens ) is the main livestock species that inhabits the Qinghai-Tibet Plateau (QTP) at regions located at high-altitude of 3,000-5,000 m. This work investigated the rumen bacterial community of yak that grazed on the QTP during the whole year to evaluate the relationship between the rumen bacterial community and the nutrient composition of forage plant at three stages. In this study, the diversity of the rumen prokaryotic community composition was monitored in 10 full-grazing yak in an alpine meadow of the QTP. The nutrient composition of three forage growth stages was determined: re-green stage (REGY), grassy stage (GY), and withered stage (WGY). High-throughput sequencing of bacterial 16S rRNA gene was used. The results showed that the nutritive composition of the alpine meadow changed with the seasons: crude protein (CP) (13.22%) was high in forage during REGY (spring), while neutral detergent fiber (NDF) (59.00%) was high during WGY (winter). Microbial diversity and richness were highest during REGY and the average number of operational taxonomic units from 30 samples was 4,470. The microbial composition was dominated by members of Bacteroidetes (51.82%), followed by Firmicutes (34.08%), and the relative microbial abundance changed in the three forage growth stages. Unweighted UniFrac distance PcoA showed that the bacterial community structure differed between REGY, GY, and WGY. Furthermore, taxonomic groups did not present differences regarding gender in these three stages. The rumen microbiota was enriched with functional potentials that were related to ABC transporters, the two-component system, Aminoacyl-tRNA biosynthesis, and metabolism of Purine, Pyrimidine, Starch and sucrose metabolism. Significant differences were found in the composition, diversity, and function of yak ruminal microorganisms during different forage growth stages. This indicates that microbial changes in the rumen depend on changes in the forage nutritional composition. These findings provide evidence on the rumen microbial diversity of yaks in the QTP., Competing Interests: The authors declare there are no competing interests., (©2019 Ma et al.)

This study aimed to research the effects of different dietary concentrate to forage (C:F) ratio on growth performance, rumen fermentation and bacteria diversity of barn feeding Tibetan sheep. The experiment contains fiver treatments (HS1, HS2 HS3, HS4 and HS5; n  = 8, respectively) based on dietary C:F ratios 0:100, 15:85, 30:70, 45:55, and 60:40, respectively. The ruminal bacterial community structure was investigated through high-throughput sequencing of 16S rRNA genes in V4 hypervariable region. The results showed that increasing dietary concentrate feed level from 0% to 60% exerted a positive effect on DMI, BW gain, gain rate and feed conversation ratio (F CR ) in Tibetan sheep. The increases dietary concentrate feed level elevatedNH 3 -N, propionate and valerate concentrations, whereas, reduced molar ratio of acetate to propionate (A/P ratio) ( P  < 0.05). For rumen bacterial diversity, increases in dietary concentrate content contributed to lower alpha diversity indexes including Shannon wiener, Chao1 and observed species, meanwhile, significantly increased the abundances of the phylum Bacteroidetes and the genus Prevotella&#95;1 ( P  < 0.05). In conclusion, increases dietary concentrate content improved the growth performance and Tibetan sheep fed diets of 45% concentrate obtained a better performance; the inclusion of concentrate in feed changed rumen fermentation from acetate fermentation to propionate fermentation, and improved the energy utilization efficiency of Tibetan sheep; the increased in concentrate content significantly reduced rumen bacteria diversity and changed the abundance of some core bacteria., Competing Interests: The authors declare there are no competing interests.

DNA hypermethylation in tumor suppressor genes has been reported in some cancers. The microRNA-34b/c (miR-34b/c) serves as tumor suppressors in different tumor types. To investigate the methylation status of miR-34b/c in ESCC, MALDI-TOF MS was used to quantitatively analyze the DNA methylation of 16 CpG sites within miR-34b/c in 145 ESCC samples, 60 cancer-adjacent normal (ACN) samples and 39 normal esophageal (NE) samples from the Kazakh population. Our results showed that the overall average methylation levels of miR-34b/c were significantly higher in the ESCC samples than they were in the ACN and NE samples ( P < 0.05). Furthermore, the methylation levels of CpG&#95;1.2.3, CpG&#95;9.10, CpG&#95;11.12.13, CpG&#95;14, and CpG&#95;15.16 of miR-34b/c were significantly higher in the ESCC tissues than they were in the ACN ( P < 0.05) and NE tissues ( P < 0.05). Additionally, the mean methylation levels at CpG&#95;9.10 and CpG&#95;14 were all significantly higher in the ACN samples than they were in the NE samples ( P < 0.01). Increased methylation levels of CpG&#95;9.10 and CpG&#95;11.12.13 in miR-34b/c predominantly occurred in the early stages (UICC I/II) of ESCC ( P < 0.05), and the methylation differences (moderately-poorly differentiated > well differentiated) in miR-34b/c CpG&#95;1.2.3 were significant ( P < 0.05). This is the first study reporting that the hypermethylation of miR-34b/c plays an important role in ESCC and is significantly correlated with the early stages and tumor differentiation of ESCC. The hypermethylation of miR-34b/c may promote the oncogenesis and progression of ESCC, and these findings may provide support for the future development of targeted therapies., Competing Interests: None., (IJCEP Copyright © 2019.)

Background: Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignancies. Neovascularization during tumorigenesis supplies oxygen and nutrients to proliferative tumor cells, and serves as a conduit for migration. Targeting oncogenes involved in angiogenesis is needed to treat organ-confined and locally advanced ESCC. Although the phospholipase C epsilon-1 (PLCE1) gene was originally identified as a susceptibility gene for ESCC, how PLCE1 is involved in ESCC is unclear., Methods: Matrix-assisted laser desorption ionization time-of-flight mass spectrometry were used to measure the methylation status of the PLCE1 promoter region. To validate the underlying mechanism for PLCE1 in constitutive activation of the NF-κB signaling pathway, we performed studies using in vitro and in vivo assays and samples from 368 formalin-fixed esophageal cancer tissues and 215 normal tissues with IHC using tissue microarrays and the Cancer Genome Atlas dataset., Results: We report that hypomethylation-associated up-regulation of PLCE1 expression was correlated with tumor angiogenesis and poor prognosis in ESCC cohorts. PLCE1 can activate NF-κB through phosphoinositide-phospholipase C-ε (PI-PLCε) signaling pathway. Furthermore, PLCE1 can bind p65 and IκBα proteins, promoting IκBα-S32 and p65-S536 phosphorylation. Consequently, phosphorylated IκBα promotes nuclear translocation of p50/p65 and p65, as a transcription factor, can bind vascular endothelial growth factor-C and bcl-2 promoters, enhancing angiogenesis and inhibiting apoptosis in vitro. Moreover, xenograft tumors in nude mice proved that PLCE1 can induce angiogenesis, inhibit apoptosis, and increase tumor aggressiveness via the NF-κB signaling pathway in vivo., Conclusions: Our findings not only provide evidence that hypomethylation-induced PLCE1 confers angiogenesis and proliferation in ESCC by activating PI-PLCε-NF-κB signaling pathway and VEGF-C/Bcl-2 expression, but also suggest that modulation of PLCE1 by epigenetic modification or a selective inhibitor may be a promising therapeutic approach for the treatment of ESCC.

Purpose:: To investigate the protective mechanisms of propofol (Pro) on renal ischemia/reperfusion (I/R) injury by studying its impact on renal I/R endoplasmic reticulum stress., Methods:: Eighteen male Sprague-Dawley rats (SD rats) were randomly divided into three groups: the I/R group, the Pro pretreatment group, and the control group, and corresponding treatments were performed. The levels of serum creatinine (Cr) and blood urea nitrogen (BUN) of each group were detected. The expression levels of CCAAT-enhancer-binding protein (C/EBP) homology protein (CHOP) and caspase-12 protein within renal tissue samples were detected by western blot., Results:: The periodic acid-Schiff (PAS) staining was performed to observe the morphological changes within the renal tissues, and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay was performed to detect the presence of renal apoptosis. The Pro pretreatment significantly reduced the serum Cr and BUN levels, as well as the expressions levels of CHOP and caspase-12 protein inside the kidney of I/R rats, improving renal pathological injury and reducing the I/R-induced renal apoptosis., Conclusion:: Propofol could downregulate the expression of stress-apoptotic proteins CHOP and caspase-12 in the endoplasmic reticulum, thus reducing renal I/R injury.

Real-time quantitative polymerase chain reaction (RT-qPCR) is a sensitive technique for gene expression studies. However, choosing the appropriate reference gene is essential to obtain reliable results for RT-qPCR assays. In the present work, the expression of eight candidate reference genes, EF1-α (elongation factor 1- α ), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), UBC (ubiquitin-conjugating enzyme), UBQ (polyubiquitin), ACT (actin), β-TUB ( β -tubulin), APT1 (adenine phosphoribosyltransferase 1), and 18S rRNA (18S ribosomal RNA), was evaluated in Achyranthes bidentata samples using two algorithms, geNorm and NormFinder. The samples were classified into groups according to developmental stages, various tissues, stresses (cold, heat, drought, NaCl), and hormone treatments (MeJA, IBA, SA). Suitable combination of reference genes for RT-qPCR normalization should be applied according to different experimental conditions. In this study, EF1-α , UBC , and ACT genes were verified as the suitable reference genes across all tested samples. To validate the suitability of the reference genes, we evaluated the relative expression of CAS , which is a gene that may be involved in phytosterol synthesis. Our results provide the foundation for gene expression analysis in A. bidentata and other species of Amaranthaceae.

Our goal was to evaluate how dental treatments under general anesthesia (GA) affect the quality of life by a prospective pair-matched design. Pediatric patients, who had received dental treatments under GA, were enrolled and were asked to complete the Early Childhood Oral Health Impact Scale (ECOHIS) before the treatment and 1 month after the treatment. To shield the observed impacts, a pair-matched control group was performed. Patients in the control group were also required to complete the ECOHIS at these different points in time. In both groups, the items of troubled sleep and oral/dental pain scored highest, whereas avoiding smiling or laughing and avoiding talking scored lowest before the treatment. The total mean score in the 2 groups was 13.1 and 13.7, respectively, and there was no significant statistical difference (P > 0.05). However, the total mean score was 1.9 in the experimental group after the treatment and smaller compared with the control group (1.9 vs. 4.7, P < 0.001). The majority of the items in both groups had an apparent effect size and the total mean effect in the experimental group was greater than that in the control group (85.5% vs. 65.7%, P < 0.001). Therefore, dental treatment under GA could provide better quality of life restoration compared with treatment over multiple visits., Competing Interests: The authors report no conflicts of interest.

Achyranthes bidentata is a popular perennial medicine herb used for 1000s of years in China to treat various diseases. Although this herb has multiple pharmaceutical purposes in China, no transcriptomic information has been reported for this species. In addition, the understanding of several key pathways and enzymes involved in the biosynthesis of oleanolic acid and ecdysterone, two pharmacologically active classes of metabolites and major chemical constituents of A. bidentata root extracts, is limited. The aim of the present study was to characterize the transcriptome profile of the roots and leaves of A. bidentata to uncover the biosynthetic and transport mechanisms of the active components. In this study, we identified 100,987 transcripts, with an average length of 1146.8 base pairs. A total of 31,634 (31.33%) unigenes were annotated, and 12,762 unigenes were mapped to 303 pathways according to the Kyoto Encyclopedia of Genes and Genomes pathway database. Moreover, we identified a total of 260 oleanolic acid and ecdysterone genes encoding biosynthetic enzymes. Furthermore, the key enzymes involved in the oleanolic acid and ecdysterone synthesis pathways were analyzed using quantitative real-time polymerase chain reaction, revealing that the roots expressed these enzymes to a greater extent than the leaves. In addition, we identified 85 ATP-binding cassette transporters, some of which might be involved in the translocation of secondary metabolites.

Objective: To investigate the effect of dexamethasone (DEX) on the microglia activation induced by glutamic acid (GLU) in rats., Methods: We isolated and cultured the microglia from the spinal cord of SD infant rats in vitro. The cell purity was tested by immunofluorescence technique. The cells were then randomly divided into 5 groups: Dulbecco' s phosphate buffered saline treatment (DPBS group); GLU treatment (GLU group); DEX pretreatment and then GLU stimulation; simultaneous treatment of DEX and GLU; GLU stimulation followed by DEX treatment. Finally, immunofluorescence technique was used to investigate the expressions of glucocorticoid receptor (GR) and CD11b/c protein., Results: Compared with the DPBS group, GLU group presented the increased expression of CD11b/c protein, the shorten length of cell processes as well as cell shape turning round. Furthermore, compared with the GLU group, the CD11b/c protein expression significantly decreased in the group treated simultaneously with DEX and GLU and the group treated with DEX after GLU stimulation. However, the expression was not different between the GLU group and the group treated with DEX and then stimulated by GLU., Conclusion: DEX could inhibit microglia activation induced by GLU, while DEX pretreatment have no such an effect on microglia.

3-hydroxypropionate (3HP) is an important platform chemical, and its biosynthesis is severely restricted by the toxicity of 3HP on cell growth and survival. To improve Escherichia coli resistance to 3HP and reduce the total production cost in industrial applications, we have identified variations in protein expression level exposed to sub-lethal concentration of this chemical using 2-dimensional gel electrophoresis. Under 3HP stress, the amount of 46 proteins was increased while the amount of 23 proteins was reduced. According to the proteomic results, overexpression of some identified proteins significantly increased the E. coli survival rate under 3HP stress. This study shed light on clues for developing E. coli strains with higher resistance to 3HP toxicity and lower production cost for industrial applications.

PMADS3, a known MADS-box transcriptional factor and a C-class gene for floral development, plays dual roles in controlling the identity of inner floral organs and the termination of flower meristems in petunia. In this study, it was confirmed by bimolecular fluorescence complementation (BiFC) assays that the PMADS3 protein can interact individually with E-class proteins FBP2, FBP5, FBP9 and PMADS12. A yeast two-hybrid cDNA library was screened using the entire PMADS3 as bait, and this identified further potential interaction candidates. Two novel genes, PheIF3f and PhAGO10, were isolated, and suggested to regulate mRNA and translational processes according to the analysis of protein functional domains and subcellular localization predictions. Notably, the PhAGO10 protein belongs to the Argonaute family, members of which are major players in small-RNA-guided gene silencing processes via mRNA cleavage or translational inhibition. The results of yeast two-hybrid and BiFC assays indicated that PheIF3f and PhAGO10 could interact with PMADS3. Our findings indicate that the C-class gene PMADS3 potentially participates in post-transcriptional control, as well as transcriptional regulation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Laparoscopic cholecystectomy is performed with increasing frequency in aging populations. However, in elderly patients, cognitive dysfunction following surgery may impair the outcome of surgical procedures. Dexmedetomidine (DEX) has been demonstrated to have a neuroprotectve effect in animal experiments. However, it is unclear whether DEX also has a neuroprotective effect in human patients. The present study was a randomized, placebo-controlled double-blind trial of 126 patients who had undergone laparoscopic cholecystectomy, using clinical interviews to determine whether intravenously administrated DEX during general anesthesia ameliorates cognitive function impairment. The cognitive deficit of each patient was assessed using the Mini-Mental State Examination (MMSE). The scores on the MMSE for the DEX and control groups one week after surgery (DEX group, 27.6±1.2; control group, 25.7±1.5) were significantly different (P=0.005). The MMSE scores of patients ≤65 years old were significantly higher than those of patients >65 one week after surgery. The MMSE scores were significantly different between the two age groups in the control patients (≤65 years old, 28.3±1.2; >65 years old, 26.6±2.1; P=0.036), while the difference was not statistically significant in the DEX-treated patients. Eight patients in the DEX group and 15 patients in the control group had mild cognitive impairment (26≥ MMSE score ≥21) although the difference was not statistically significant. The findings of the present study support the hypothesis that DEX administration may be an effective method for ameliorating postoperative cognitive impairment in elderly patients who have undergone laparoscopic cholecystectomy. Further research is required to confirm the findings of the present study.

Pain in the cervical region of air force pilots, who are exposed to high G-forces, is a specifically occupational health problem. To minimize neck problems, the cervical muscles need specific strength exercise. It is important that the training for the neck must be carried out with optimal resistance in exercises. The model NSE-1 neck training machine for pilots was designed for neck strengthening exercises under safe and effective conditions. In order to realize the functions of changeable velocity and resistant (CVR) training and neck isometric contractive exercises, the techniques of adaptive hydraulics, sensor, optic and auditory biological feedback, and signal processing were applied to this machine. The training system mainly consists of mechanical parts (including the chair of flexion and extension, the chair of right and left lateral flexion, the components of hydraulics and torque transformer, etc.), and the software of signal processing and biological feedback. Eleven volunteers were selected for the experiments of neck isometric contractive exercises, three times a week for 6 weeks, where CVR training (flexion, extension, right, left lateral flexion) one time a week. The increase in relative strength of the neck (flexion, extension, left and right lateral flexion) was 70.8%, 83.7%, 78.6% and 75.2%, respectively after training. Results show that the strength of the neck can be increased safely, effectively and rapidly with NSE-1 neck training machine to perform neck training.