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Flowering time is an important agronomic trait of Chinese cabbage with late flowering being a primary breeding objective. In our previous work, we obtained Chinese cabbage-cabbage translocation lines that contained several beneficial cabbage genes. Cabbage-specific molecular markers show that these genes were coming from chromosome C01 of cabbage. In this study, we investigated the inheritance of flowering time in a couple of translocation lines and analyzed the transmission rate of molecular markers in the offspring. Consequently, we obtained the late flowering Chinese cabbage-cabbage translocation line ‘AT7-4’ in which the flowering time was later than that of ‘85-1’ by about 7 days under 4-week vernalization. Based on previous studies of the genomes of Chinese cabbage and cabbage, we located the cabbage-specific molecular markers that were closely linked at the top of the chromosome A01 in the F2 mapping population generated by self-crossing F1s derived from a cross between the translocation line ‘AT7-4’ and Chinese cabbage ‘14-36’. Five flowering-related genes in the alien fragment were found by functional annotation and their molecular markers were developed. This study lays the foundation for the future improvement of Chinese cabbage varieties using A-C translocation lines.

According to the Leadership Efficacy Model, leadership efficacy depends on leaders’ tendency to make linear relationships between leadership philosophy, practice, and criteria (i.e., congruence of leadership cycles). Moreover, efficacy increases if coaches make these linear relationships by using the optimal leadership profile and by considering the antecedent factors of leadership (characteristics of the leader, team members, and organizational conditions; i.e., favorability of conditions for leadership). This study compared the perceptions of athletes and their coaches regarding leadership cycles, and tested the moderator role of optimal leadership profile and leadership favorability in the relationship between leadership cycles and leadership efficacy. This study included 92 football athletes (ages less than 17 and 19 years) and respective coaches (n = 5). The evaluation protocol included measures of leadership cycles, leadership styles, leadership favorability, and sport performance perception. Athletes and coaches agreed on coaches’ need to increase their practice and criteria, but athletes also considered that coaches should better clarify their philosophy. Regression analyses confirmed that leadership congruency predicts higher perceptions of team performance in athletes. Moreover, optimal leadership profile and higher leadership favorability were associated with higher team and individual performance. However, these two factors did not moderate the relationship between leadership congruency and efficacy.

Paleopalynology evidence and megafossil records indicated that tree fern order Cyatheales, played an important role in Mesozoic terrestrial ecosystem. Few fossil records of the family Thyrsopteridaceae have been reported so far. In the present study, we describe a distinctive fertile pinnule segment of a fern plant preserved in a mid-Cretaceous amber from Myanmar. The well-preserved fertile pinnule is identified as a new species of the extant tree fern genus Thyrsopteris (Thyrsopteridaceae): Thyrsopteris cyathindusia n. sp. It represents the second fossil species of Thyrsopteridaceae found in the mid-Cretaceous amber from Myanmar. The new species shows distinctive characteristics of the sporophyll that are considered paleoecologically/paleobiogeographically significant for evolution of the Thyrsopteridaceae.

Colorectal cancer (CRC) is the third leading cancer globally. Metagenomics has been widely used to analyze the association between the gut microbiota and CRC based on bacterial genus- or species-level comparisons, providing evidence of dysbiosis in CRC development. However, this kind of analysis is unable to provide strain-level information for understanding the individual role of a species in CRC. Here, we used culturomics to isolate CRC mucosal samples and selected 158 Escherichia coli strains to reveal their differences in both genomics and functions by means of phylogenetic analysis and inflammatory induction based on cell and animal experiments. Through genomic comparison, these strains were divided into five phylogroups. The representative strains of each phylogroup significantly induced different levels of cytokine secretion by human leukemic monocyte (THP-1 cell)-based Transwell and animal experiments. Further bioinformatic analysis revealed different profiles of single-nucleotide polymorphisms, genes, and metabolic pathways in the different phylogroups, which can improve the current understanding of the phenotypic differences between these strains. The strain differences revealed in both genomics and functions indicate that the microbiota’s function at the strain level should be investigated in order to understand the interacting mechanisms between hosts and gut bacteria.

The availability of public genomic resources can greatly assist biodiversity assessment, conservation, and restoration efforts by providing evidence for scientifically informed management decisions. Here we survey the main approaches and applications in biodiversity and conservation genomics, considering practical factors, such as cost, time, prerequisite skills, and current shortcomings of applications. Most approaches perform best in combination with reference genomes from the target species or closely related species. We review case studies to illustrate how reference genomes can facilitate biodiversity research and conservation across the tree of life. We conclude that the time is ripe to view reference genomes as fundamental resources and to integrate their use as a best practice in conservation genomics., Trends in Genetics, 39 (7), ISSN:0168-9525

Tiger pufferfish (Takifugu rubripes) is one of Asia's most economically valuable aquaculture species. However, winter production of this species in North China is limited by low water temperature and unavailability of high-quality feed, resulting in high mortality and low profitability. Therefore, the aim of this study was to evaluate the effect of feeding frequency (F1: one daily meal; F2: two daily meals; F3: four daily meals; F4: continuous diurnal feeding using a belt feeder) on the growth performance, plasma biochemistry, digestive and antioxidant enzyme activities, and expression of appetite-related genes in T. rubripes (initial weight: 266.80 ± 12.32 g) cultured during winter (18.0 ± 1.0 °C) for 60 days. The results showed that fish in the F3 group had the highest final weight, weight gain rate, specific growth rate, survival rate, and best feed conversion ratio. Additionally, daily feed intake increased significantly with increasing feeding frequency. The protein efficiency and lipid efficiency ratios of fish in the F3 group were significantly higher than those of fish in the other groups. Furthermore, total cholesterol, triglycerides, and glucose levels increased with increasing feeding frequency, peaking in the F2 group and decreasing under higher feeding frequencies. The antioxidant (superoxide dismutase, catalase, glutathione, and glutathione peroxidase) and digestive (trypsin, amylase, and lipase) enzyme activities of fish in the F1 group were significantly higher than those of fish in the F3 and F4 groups. Additionally, there was a decrease in orexin expression with increasing feeding frequency. In contrast, the expression levels of tachykinin, cholecystokinin, and leptin increased with increasing feeding frequency, peaking in the F4 group. Overall, the findings of this study indicated that a feeding frequency of four meals per day was optimal for improved growth performance of pufferfish juveniles cultured during winter.

Programmed cell death (PCD) is an important pathologic process after spinal cord injury (SCI), and as a newly type of PCD, ferroptosis is also involved in the secondary SCI, however, the underlying molecular mechanisms remain unclear. Integrating animal experiment and bioinformatics, we validated the ferroptotic phenotype in SCI first, and then bioinformatic analyses, including Gene Ontology enrichment analysis, Kyoto Encyclopedia of Genes and Genomes pathway analysis, gene set enrichment analysis and protein-protein interaction analysis were performed to investigate the ferroptotic genes at 1 day, 3 days, 7 days, 14 days and 56 days post-SCI, finally, the ferroptotic genes in SCI were identified and expression of 5 key genes were validated by western blot. The ferroptotic symbols including iron overload, lipid peroxidation, shrunken mitochondria and ROS accumulation were detected in the acute and sub-acute phase of SCI. The outcomes of bioinformatics suggested that mTOR signaling pathway, HIF-1 signaling pathway, VEGF signaling pathway, Protein processing in endoplasmic reticulum were involved in ferroptotic regulation and ATF-3, XBP-1, HO-1, DDIT-3 and CHAC-1 were selected as the ferroptotic key genes in SCI. Besides, response to oxidative stress, amide metabolic process, cation transport and cytokine production were showed as the essential biological process in ferroptosis after SCI. The ferroptotic phenotype following SCI was validated and the ferroptotic genes and signaling pathways were identified. The results contribute to exploring the ferroptotic mechanism underlying secondary SCI and to providing potential target for clinical treatment.

The histone demethylase KDM6A controls gene expression by the epigenetic regulation of H3K27 methylation and functions in diverse processes, including differentiation, development, and cancer. Here, we investigated the role of KDM6A in prostate cancer. Specific deletion of KDM6A in the adult mouse prostate epithelium strongly inhibited tumor progression initiated by the loss of PTEN. Mechanistically, KDM6A promoted prostate tumorigenesis and lipid metabolism by binding to the SREBP1c promoter to increase SREBP1c transcription. USP7 deubiquitinated KDM6A to increase its expression. KDM6A was significantly upregulated in prostate cancer and positively associated with USP7 expression. Furthermore, targeting KDM6A stability by inhibiting USP7 in conditional knockout mice and xenograft models markedly suppressed prostate cancer growth and significantly enhanced KDM6A inhibitor efficacy. Collectively, these findings indicate that KDM6A regulates prostate lipid metabolism and is essential for prostate tumorigenesis initiated by PTEN loss. Targeting USP7/KDM6A could be a valuable strategy to ameliorate prostate cancer progression and therapeutic resistance. Significance: These findings show that KDM6A supports prostate tumorigenesis by promoting SREBP1c-mediated lipid metabolism, proposing targeting the USP7/KDM6A axis as a therapeutic strategy for treating prostate cancer.

Brassica crops, which are of worldwide importance, provide various oil, vegetable and ornamental products, as well as feedstocks for animal husbandry and biofuel industry. Cinnamoyl-CoA reductase (CCR) is the entry point to the lignin pathway and a crucial locus in manipulation of associated traits, but CCR-associated metabolism and traits in Brassica crops have remained largely unstudied except in Arabidopsis thaliana. We report the identification of 16 CCR genes from Brassica napus and its parental species B. rapa and B. oleracea. The BnCCR1 and BnCCR2 subfamilies displayed divergent organ-specificity and participation in the yellow-seed trait. Their functions were dissected via overexpression of representative paralogs in B. napus. BnCCR1 was expressed preferentially in G- and H-lignin biosynthesis and vascular development, while BnCCR2 was expressed in S-lignin biosynthesis and interfascicular fiber development. BnCCR1 showed stronger effects on lignification-related development, lodging resistance, phenylpropanoid flux control, and seed coat pigmentation, whereas BnCCR2 showed a stronger effect on sinapate biosynthesis. BnCCR1 upregulation delayed bolting and flowering time, while BnCCR2 upregulation weakened the leaf vascular system in consequence of suppressed G-lignin accumulation. BnCCR1 and BnCCR2 were closely but almost oppositely linked with glucosinolate metabolism via inter-pathway crosstalk. We conclude that BnCCR1 and BnCCR2 subfamilies offer great but differing potential for manipulating traits associated with phenylpropanoids and glucosinolates. This study reveals the CCR1–CCR2 divergence in Brassicaceae and offers a resource for rapeseed breeding for lodging resistance, yellow-seed traits, and glucosinolate traits.

The age of the Silurian “Lower Red Beds” (LRBs), which occur widely in South China, has long been controversial because of the scarcity of index fossils. Here, we report a fauna with abundant trilobites from the LRBs in Wuhan, Middle Yangtze region, consisting of Aulacopleura (Paraaulacopleura) pijiazhaiensis, ‘Encrinuroides’ yichangensis, Gaotania hubeiensis, Hyrokybe gaotanensis, Japonoscutellum yichangense, Latiproetus latilimbatus, and Ptilillaenus lojopingensis. These trilobites are common in the Leijiatun, upper Xiangshuyuan, Shihniulan, and Lojoping formations in the Upper Yangtze region, and belong to the middle to upper Aeronian Ptilillaenus or ‘Encrinuroides’–Ptilillaenus–Latiproetus assemblages. The discovery of this trilobite assemblage suggests that the LRBs are probably partly of middle–late Aeronian age in the Middle Yangtze region. Thus, the lower boundary of the LRBs is likely diachronous in the Yangtze region, ranging from the middle–late Aeronian to the early Telychian. The discovery of the Ptilillaenus assemblage in the Wuhan LRBs expands the paleogeographic distribution of this assemblage and provides evidence of deeper depositional environment of the LRBs in the study area versus shallow waters in other areas of the Yangtze region.

Peanut (Arachis hypogea L.) bacterial wilt (BW) is a devastating disease caused by Ralstonia solanacearum that results in severe yield and quality losses. Plant defensins are short cysteine-rich peptides with antimicrobial activity. The role of defensin genes (AhDef) in peanut is unclear. A genome-wide investigation of AhDef genes was undertaken, and 12 identified AhDef genes were classified into two groups containing the gamma-thionin domain formed by four disulfide pairs: Cys1-Cys8, Cys2-Cys5, Cys3-Cys6, and Cys4-Cys7. In silico analysis revealed that AhDef genes showed highly conserved architectural features and contained cis-elements associated with phytohormone signaling and defense responses. A highly resistant cultivar, H108 (R) and a susceptible accession, H107 (S) were tested by R. solanacearum inoculation. H108 (R) showed fewer symptoms than H107 (S) owing to inhibition of bacterial reproduction and spread in the vascular bundles of roots and stems. In a transcriptomic expression profile, AhDef genes, particularly AhDef1.6 and AhDef2.2, were up-regulated in H108 (R) compared with H107 (S) under R. solanacearum infection and phytohormone treatment. Subcellular localization showed that the AhDef1.6 and AhDef2.2 proteins were both expressed specifically on the plasma membrane. Overexpression of protein fusion AhDef2.2-YFP in Nicotiana benthamiana and peanut leaves increased resistance to R. solanacearum, suggesting its role in response to BW infection. AhDef2.2 may be valuable for peanut resistance breeding.

Straw strip mulching (SM) is a new mulching technology. From 2012 to 2018, SM’s effects on soil moisture and temperature and production performances were compared with other mulching practices, using three treatments: full-cover plastic mulch (PM), no mulch with wheat sown in rows as the control (CK), and SM with 50% to 59% of the field area mulched. Compared with CK, on average over six growing seasons, SM and PM increased grain yield by 27.0% and 21.7%, straw yield by 21.6% and 22.6%, kernels ha−1 by 26.6% and 19.0%, net income by 29.8% and −25.0%, soil temperature at 5 cm by −1.5 °C and 0.2 °C from overwintering to maturity, and soil water storage at 0 to 200 cm by 25 and 22 mm, respectively. The increase in soil moisture in SM and PM was greater in the early period (overwintering to jointing) than in the later period (booting to maturity) and at 0 to 120 cm than at 120 to 200 cm in the early period. Although the mean evapotranspiration of whole growth period across six seasons was similar among treatments, SM and PM increased water consumption during the key formation period of yield components after overwintering by 16 and 32 mm, respectively, while reducing it before overwintering. Compared with CK, SM and PM had the effects of warming during overwintering and cooling after jointing. By increasing water consumption after overwintering and ratio of transpiration to evapotranspiration and providing favorable soil temperature for multiple growth stages and more sufficient soil moisture, SM and PM promoted vegetative growth and increased kernels ha−1, the main mechanisms by which SM and PM increased grain yield relative to CK. Relative to PM, SM is a more economically beneficial and environment-friendly technology for dryland wheat production.

The outer membrane protein (ExbB) of Pseudomonas fluorescens present in the outermost layer may possess immune functions and potential applications in subunit vaccines. ExbB bioinformatic analysis elucidated that the relationship among Pseudomonas species was closer than others, and anti-ExbB serum might provide cross-protective ability to resist Pseudomonas bacterial infection in animals. ExbB was obtained via molecular cloning, prokaryotic expression, and purification. The optimal expression conditions were a strain OD600 value of 1.0 and IPTG concentration of 0.3 mmol/L, inducing time of 3 h, and inducing temperature of 32℃. Mice immunized to ExbB could significantly increase (p

An unacceptable increase in antibacterial resistance has arisen due to the abuse of multiple classes of broad-spectrum antibiotics. Therefore, it is significant to develop new antibacterial agents, especially those that can accurately identify and kill specific bacteria. Herein, we demonstrate a kind of perilla-derived carbon nanodots (CNDs), integrating intrinsic advantages of luminescence and photodynamic, providing the opportunity to accurately identify and kill specific bacteria. The CNDs have an exotic-doped and π-conjugated core, vitalizing them near-infrared (NIR) absorption and emission properties with photoluminescence quantum yield of 21.1%; hydrophobic chains onto the surface of the CNDs make them to selectively stain Gram-positive bacteria by insertion into their membranes. Due to the strong absorption in NIR region, reactive oxygen species are in situ generated by the CNDs onto bacterial membranes under 660 nm irradiation, and 99.99% inactivation efficiency against Gram-positive bacteria within 5 min can be achieved. In vivo results demonstrate that the CNDs with photodynamic antibacterial property can eliminate the inflammation of the area affected by methicillin-resistant Staphylococcus aureus (MRSA), and enabling the wound to be cured quickly.

BACKGROUND: Pulmonary metastasis is the most frequent cause of death in osteosarcoma (OS) patients. Recently, several bioinformatics studies specific to pulmonary metastatic osteosarcoma (PMOS) have been applied to identify genetic alterations. However, the interpretation and reliability of the results obtained were limited for the independent database analysis. OBJECTIVE: The expression profiles and key pathways specific to PMOS remain to be comprehensively explored. Therefore, in our study, three original datasets of GEO database were selected. METHODS: Initially, three microarray datasets (GSE14359, GSE14827, and GSE85537) were downloaded from the GEO database. Differentially expressed genes (DEGs) between PMOS and nonmetastatic osteosarcoma (NMOS) were identified and mined using DAVID. Subsequently, GO and KEGG pathway analyses were carried out for DEGs. Corresponding PPI network of DEGs was constructed based on the data collected from STRING datasets. The network was visualized with Cytoscape software, and ten hub genes were selected from the network. Finally, survival analysis of these hub genes also used the TARGET database. RESULTS: In total, 569 upregulated and 1238 downregulated genes were filtered as DEGs between PMOS and NMOS. Based on the GO analysis result, these DEGs were significantly enriched in the anatomical structure development, extracellular matrix, biological adhesion, and cell adhesion terms. Based on the KEGG pathway analysis result, these DEGs were mainly enriched in the pathways in cancer, PI3K-Akt signaling, MAPK signaling, focal adhesion, cytokine-cytokine receptor interaction, and IL-17 signaling. Hub genes (ANXA1 and CXCL12) were significantly associated with overall survival time in OS patient. CONCLUSION: Our results may provide new insight into pulmonary metastasis of OS. However, experimental studies remain necessary to elucidate the biological function and mechanism underlying PMOS.

Delimiting species requires multiple sources of evidence. Here, we delimited two varieties of Halenia elliptica (Gentianaceae) using several lines of evidence, including morphological traits and mating system in a sympatric population, phylogenetic relationships based on nrITS and cpDNA (rpl16) data, and complete chloroplast genome sequences. Comparative analysis of 21 morphological traits clearly separates the two varieties of H. elliptica. Examination of the flowering process and pollination treatments indicate that H. elliptica var. grandiflora produces seeds via outcrossing, whereas H. elliptica var. elliptica produces seeds via mixed mating. Furthermore, hand-pollinated hybridization of the two varieties produced no seeds. Observations of pollinators showed that when bees began a pollination bout on H. elliptica var. grandiflora they preferred to continue pollinating this variety; however, when they began a pollination bout on H. elliptica var. elliptica, they showed no preference for either variety. Phylogenetic analysis confirmed the monophyly of H. elliptica, which was further divided into two monophyletic clades corresponding to the two varieties. A large number of variants from the chloroplast genomes reflected remarkable genetic dissimilarities between the two varieties of H. elliptica. We recommend that the two varieties of H. elliptica should be revised as two species (H. elliptica and Halenia grandiflora). Our findings indicate that H. elliptica varieties may have split into two separate species due to a shift in mating system, changes in flowering phenology and/or post-pollination reproductive isolation.

Understanding how natural hybridization and polyploidizations originate in plants requires identifying potential diploid ancestors. However, cryptic plant species are widespread, particularly in Ceratopteris (Pteridaceae). Identifying Ceratopteris cryptic species with different polyploidy levels is a challenge because Ceratopteris spp exhibit high degrees of phenotypic plasticity. Here, two new cryptic species of Ceratopteris, Ceratopteris chunii and C. chingii, are described and illustrated. Phylogenetic analyses reveal that each of the new species form a well-supported clade. Ceratopteris chunii and C. chingii are similar to C. gaudichaudii var. vulgaris and C. pteridoides, respectively, but distinct from their relatives in the stipe, basal pinna of the sterile leaf or subelliptic shape of the fertile leaf, as well as the spore surface. In addition, chromosome studies indicate that Ceratopteris chunii and C. chingii are both diploid. These findings will help us further understand the origin of Ceratopteris polyploids in Asia.

Introduction Metagenomic next-generation sequencing (mNGS) assay for detecting infectious agents is now in the stage of being translated into clinical practice. With no approved approaches or guidelines available, laboratories adopt customized mNGS assays to detect clinical samples. However, the accuracy, reliability, and problems of these routinely implemented assays are not clear. Objectives To evaluate the performance of 90 mNGS laboratories under routine testing conditions through analyzing identical samples. Methods Eleven microbial communities were generated using 15 quantitative microbial suspensions. They were used as reference materials to evaluate the false negatives and false positives of participating mNGS protocols, as well as the ability to distinguish genetically similar organisms and to identify true pathogens from other microbes based on fictitious case reports. Results High interlaboratory variability was found in the identification and the quantitative reads per million reads (RPM) values of each microbe in the samples, especially when testing microbes present at low concentrations (1 × 103 cell/ml or less). 42.2% (38/90) of the laboratories reported unexpected microbes (i.e. false positive problem). Only 56.7% (51/90) to 83.3% (75/90) of the laboratories showed a sufficient ability to obtain clear etiological diagnoses for three simulated cases combined with patient information. The analysis of the performance of mNGS in distinguishing genetically similar organisms in three samples revealed that only 56.6% to 63.0% of the laboratories recovered RPM ratios (RPMS. aureus/RPMS. epidermidis) within the range of a 2-fold change of the initial input ratios (indicating a relatively low level of bias). Conclusion The high interlaboratory variability found in both identifying microbes and distinguishing true pathogens emphasizes the urgent need for improving the accuracy and comparability of the results generated across different mNGS laboratories, especially in the detection of low-microbial-biomass samples.

Localized corrosion of metallic biomaterials is a concerning problem for implant applications. In the present work, the interaction between localized corrosion of Ti6Al4V implant material and adsorption of bovine serum albumin (BSA) was investigated by electrochemical tests, surface morphological and compositional analysis. The artificial crevice was set up on the Ti6Al4V surface to simulate the crevice corrosion of metallic implants. The results show that the BSA adsorption is regulated by the heterogeneous metallic ion release resulted from the crevice corrosion. The negatively charged BSA prefers to aggregate at the interior of artificial crevice due to electrostatic interaction with the concentrated metallic cations in the crevice. In return, the non-uniform BSA adsorption can decrease the crevice corrosion susceptibility of the Ti6Al4V implant material, even though the presence of BSA accelerates the dissolution of passive films. For the Ti6Al4V alloys with artificial crevice, stable localized corrosion occurs after 96 h of immersion in the PBS free of BSA while no stable localized corrosion can be determined in the presence of 4% BSA. It is deduced that the presence of BSA can reduce the surface potential discrepancy between the crevice interior and the crevice exterior. The disclosed mechanism of albumin-corrosion interaction is vital to the localized corrosion control of metallic implants in clinical use.

Clustered regularly interspaced short palindromic repeats (CRISPR) technology emerges a remarkable potential for cure of refractory cancer like metastatic breast cancer. However, how to efficiently deliver the CRISPR system with non-viral carrier remains a major issue to be solved. Here, we report a kind of targeted core-shell nanoparticles (NPs) carrying dual plasmids (pHR-pCas9) for precise CCCTC-binding factor (CTCF) gene insert to circumvent metastatic breast cancer. The targeted core-shell NPs carrying pHR-pCas9 can accomplish γGTP-mediated cellular uptake and endosomal escape, facilitate the precise insert and stable expression of CTCF gene, inhibit the migration, metastasis, and colonization of metastatic breast cancer cells. Besides, the finding further reveals that the inhibitory mechanism of metastasis could be associated with up-regulating CTCF protein, followed by down-regulating stomatin (STOM) protein. The study offers a universal nanostrategy enabling the robust non-viral delivery of gene-editing system for treatment of severe illness.

Circulating tumor cells (CTCs), as important liquid biopsy target, can provide valuable information for cancer progress monitoring and individualized treatment. However, current isolation platforms incapable of balancing capture efficiency, specificity, cell viability, and gentle release have restricted the clinical applications of CTCs. Herein, inspired by the structure and functional merits of natural membrane interfaces, we established an antibody-engineered red blood cell (RBC-Ab) affinity interface on microfluidic chip for high-performance isolation and release of CTCs. The lateral fluidity, pliability, and anti-adhesion property of the RBC microfluidic interface enabled efficient CTCs capture (96.5%), high CTCs viability (96.1%), and high CTCs purity (average 4.2-log depletion of leukocytes). More importantly, selective lysis of RBCs by simply changing the salt concentration was utilized to destroy the affinity interface for efficient and gentle release of CTCs without nucleic acid contamination. Using this chip, CTCs were successfully detected in colon cancer samples with 90% sensitivity and 100% specificity (20 patients and 10 healthy individuals). After the release process, KRAS gene mutations of CTCs were identified from all the 5 cancer samples, which was consistent with the results of tissue biopsy. We expect this RBC interface strategy will inspire further biomimetic interface construction for rare cell analysis.

Zika virus (ZIKV), a flavivirus associated with neurological disorders, constitutes a global health threat. During pregnancy, ZIKV traverses the placenta and causes congenital disease such as microcephaly and Guillain-Barre syndrome in newborns. To develop a specific antiviral therapy against ZIKV-induced microcephaly that could cross placental and blood-brain barriers, we designed targeted small extracellular vesicles (sEVs) encapsulating antiviral siRNA (small interfering RNA) to inhibit ZIKV. The neuro-specific targeting was achieved by engineering EVs membrane protein lamp2b fused with a neuron-specific rabies virus glycoprotein derived peptide (RVG). Intravenous administration of the RVG-engineered sEVs loaded with siRNA (ZIKV-specific siRNA) protected pregnant AG6 mice against vertical transmission of ZIKV. Particularly, sEVsRVG-siRNA traversed placental and blood-brain barriers and suppressed ZIKV infection in fetal brains. Moreover, sEVsRVG-siRNA alleviated the neuroinflammation and neurological damage caused by ZIKV in the fetal mouse model. In general, we developed a sEVs-based targeted system of antiviral therapy for brain and fetal brain infections.

With rapid development of marine infrastructures, materials with better biocorrosion resistance and antibiofouling performance will be highly demanded. Ti6Al4V alloy is susceptible to the above. The inhibition to the microbiologically influenced corrosion of Ti6Al4V-5Cu alloy against Pseudomonas aeruginosa was investigated using antibacterial test, electrochemical techniques, surface analysis, and weight loss test conducted for 2.5 months. At a sputtering depth of 0 nm, the passive film of Ti6Al4V-5Cu alloy was mainly composed of ideal oxide TiO2. With increasing sputtering thickness to 6 nm, Ti2O3 and TiO were detected with a relative fraction of 14.6% and 14.8%, respectively, in the oxide layer of Ti6Al4V-5Cu alloy. In contrast, the outermost layer of Ti6Al4V alloy was predominantly composed of TiO2 but Ti2O3 (22.8%), Al2O3 and V2O5 were also detected. With increasing sputtering depth to 6 nm, fitting revealed the presence of Ti2O3 and TiO with relative fractions of 25.3% and 35.8%, respectively. Yet, a spot of TiO (8%) was also observed at 12 nm in the oxide layer of Ti6Al4V alloy. Although the addition of Cu into Ti6Al4V alloy generated the self-healing property of passive film in the presence of P. aeruginosa, it also reduced resistance to corrosion in general condition.

Proteomics is the study of all proteins expressed by a cell or even an organism. However, knowledge of proteins that regulate the fineness of cashmere is limited. Liaoning Cashmere goat (LCG) is a valuable genetic resource of China. The skin samples of Liaoning cashmere goats during the growing period were collected performed Tandem Mass Tag (TMT) method and identified 117 differentially expressed proteins in CT_LCG (course type) and FT_LCG (fine type). To verify protein genes differentially expressed in LCG, we performed PRM validation on three candidate proteins (ALB, SDC1 and ITGB4) in CT-LCG and FT-LCG. Furthermore, primary metabolic process and lysosome are most enriched in the GO and KEGG pathways, respectively. In addition, we also derived a protein-protein interaction (PPI) regulatory network from the perspective of bioinformatics. This study sought to elucidate the molecular mechanism of differential proteins regulating cashmere fineness of Liaoning cashmere goats by using TMT quantitative proteomics analysis. Differentially expressed proteins ALB and SDC1 may regulate cashmere fineness, ITGB4 can be further studied as a promising protein. They can be used as key genes to lay a foundation for the study of cashmere fineness of Liaoning cashmere goats.

OBJECTIVE: To explore susceptibility loci associated with uveitis in Behçet's disease (BD).METHODS: We conducted a 2-stage study, consisting of a genome-wide association study (GWAS) stage and a replication stage, in a Chinese population. The GWAS stage included 978 cases with BD-related uveitis and 4,388 controls, and the replication stage included 953 cases with BD-related uveitis and 2,129 controls. Luciferase reporter analysis and chromatin immunoprecipitation assay were performed to explore the functional role of susceptibility genetic variants near ZMIZ1.RESULTS: Three independent HLA alleles (HLA-B51 [3.75 × 10-190 ], HLA-A26 [1.50 × 10-18 ], and HLA-C0704 [3.44 × 10-16 ]) were identified as having a genome-wide association with BD-related uveitis. In the non-HLA region, in addition to confirming 7 previously reported loci, we identified 22 novel susceptibility variants located in 16 loci. Meta-analysis of the Chinese cohort consisting of 1,931 cases and 6,517 controls and a published Japanese cohort of 611 cases and 737 controls showed genome-wide significant associations with ZMIZ1, RPS6KA4, IL10RA, SIPA1-FIBP-FOSL1, and VAMP1. Functional experiments demonstrated that genetic variants of ZMIZ1 were associated with enhanced transcription activity and increased expression of ZMIZ1.CONCLUSION: This GWAS study identified a novel set of genetic variants that are associated with susceptibility to uveitis in BD. These findings enrich our understanding of the contribution of genetic factors to the disease.

Background: Syphilis is a chronic infectious disease caused by Treponema pallidum (Tp) infection, which causes local inflammation in the host. TpF1 is an oligomeric protein expressed by the Tp-infected host that can induce the host immune response. There are few studies regarding the role of TpF1 in macrophage activation and the subsequent release of cytokines. Objective: The objective of this study is to elucidate the effects of TpF1 on the pathological process of Syphilis. In addition, we explored how purinergic 2X7 (P2X7R) induced NOD-like receptor family protein 3 (NLRP3) -dependent release of interleukin-1β (IL-1β) and the underlying mechanisms. Methods: We explored the influence of TpF1 on cytokine release by macrophages using qRT-PCR and ELISA. The specific phenotype of activated macrophages was determined by flow cytometry. Results: TpF1 was able to activate macrophages and induce the M1 macrophage phenotype. Moreover, TpF1 activated the NLRP3 inflammasome in macrophages, which was mediated by P2X7R. Conclusions: The Tp-induced protein TpF1 is able to induce macrophage activation and P2X7R-induced NLRP3-dependent release of IL-1β. Our findings provide a theoretical basis for clarifying the clinical symptoms and pathogenesis of syphilis.

Follicular thyroid carcinoma (FTC) is the second most common form of thyroid malignancy, and it is associated with more aggressive growth and worse long-term survival outcomes relative to papillary thyroid carcinoma (PTC). Reliable approaches to preoperative FTC detection, however, remain to be established. Herein, a targeted Affibody-Au-Tripod nanoprobe was developed and successfully utilized to facilitate the targeted photoacoustic imaging (PAI) of epidermal growth factor receptor (EGFR)-positive cells and tumors. These Affibody-Au-Tripods were found to be highly sensitive and specific for cells expressing EGFR when used as a PA contrast agent in vitro, and studies conducted in an FTC-133 subcutaneous tumor model system in mice further revealed that these Affibody-Au-Tripods were able to specifically target these EGFR-expressing tumors while providing a strong photoacoustic signal in vivo. Importantly, these nanoprobes exhibited negligible cytotoxicity and robust chemical and physical stability, making Affibody-Au-Tripods promising candidates for targeted PAI-based FTC diagnosis. In addition, these nanoprobes have the potential to facilitate the individualized treatment of patients harboring EGFR-positive tumors.

Colorectal cancer (CRC) is still the leading cause of cancer death worldwide, but the clinical effect of drug therapy such as irinotecan is not an ideal way at present. In recent years, probiotics have attracted much attention, and the combination of probiotics may play an important role in the prevention and treatment of CRC. This work proposed a cellular chip-MS system, to study the synergistic effects of probiotic Lactobacillus rhamnosus GG (L.GG) and irinotecan on HCT116 cells by cell viability and on-line mass spectrometry (MS) analysis. The double-layer chip sandwiched with a polycarbonate membrane can co-culture HCT116 cells and L.GG. And the solid phase microextraction chip can be used for desalination and concentration. Finally, the extracted chemicals were entered the electrospray ionization quadrupole time-of-flight MS to detect irinotecan metabolites. The results showed that with the increasing concentration of co-cultured L.GG, the percentage of living HCT116 cells decreased, but the relative amount of metabolized SN-38 by HCT116 cells increased. Therefore, the microfluidic system can be used to detect and monitor the synergistic effect of irinotecan-L.GG combination on HCT116 cells. In summary, our study provided experimental evidence for the first time with potential applications of irinotecan-L.GG combination in CRC treatment, and the cellular chip-MS system as a powerful tool can be used in the experiments of probiotics as new drugs.

T cell engaged bispecific antibody (TCB) is an effective immunotherapy for cancer treatment. Through co-targeting CD3 and tumor-associated antigen (TAA), TCB can redirect CD3+ T cells to eliminate tumor cells regardless of the specificity of T cell receptor. Tissue factor (TF) is a TAA that involved in tumor progression. Here, we designed and characterized a novel TCB targeting TF (TF-TCB) for the treatment of TF-positive tumors. In vitro, robust T cell activation, tumor cell lysis and T cell proliferation were induced by TF-TCB. The tumor cell lysis activity was dependent upon both CD3 and TF binding moieties of the TF-TCB, and was related to TF expression level of tumor cells. In vivo, in both tumor cell/human peripheral blood mononuclear cells (PBMC) co-grafting model and established tumor models with poor T cell infiltration, tumor growth was strongly inhibited by TF-TCB. T cell infiltration into tumors was induced during the treatment. Furthermore, efficacy of TF-TCB was further improved by combination with immune checkpoint inhibitors. For the first time, our results validated the feasibility of using TF as a target for TCB and highlighted the potential for TF-TCB to demonstrate efficacy in solid tumor treatment.

OBJECTIVES This study aimed to investigate the relationship between pre-procedure high-sensitivity C-reactive protein (hsCRP), free fatty acid (FFA), and uric acid (UA) levels and post-procedure mortality and morbidity of endovascular revascularization of arterial femoropopliteal occlusion lesions. METHODS This was a retrospective review of clinical data retrieved from a prospectively held database in Peking Union Medical College Hospital. 71 Patients who underwent endovascular treatment (EVT) for femoropopliteal occlusive disease between January 1, 2014 and November 1, 2017, were included in this study. Endpoints were defined as major adverse limb events (MALE; target vessel revascularization, amputation, or disease progression) and major adverse cardiovascular events (MACE; stroke, myocardial infarction, or all-cause death) during the entire follow-up period. Univariate and multivariate Cox proportional hazards regression models were used to evaluate the relationship of elevated biomarker levels (hsCRP, FFA and UA, measured by immunoturbidimetry assay, enzymatic assay and enzymatic assay, respectively) to MALE and MACE outcomes. RESULTS Seventy-one patients (72 limbs) with sufficient follow-up information were included in the analysis. The mean age was 69.7±8.6 years; 21.1% were female. The Rutherford class of target limbs were ≥ 3. The median follow-up was 36 (range 18-59 months). Univariate analyses revealed that patients with elevated hsCRP levels had an increased risk of MALE (hazard ratio [HR], 2.682; 95% confidence interval [CI], 1.281-5.617, P=0.009). High FFA levels were associated with an increased risk of MALE (HR, 2.658; 95% CI, 1.075-6.573; P=0.034). Multivariate analyses demonstrated that elevated hsCRP values (HR, 4.015; 95% CI, 1.628-10.551; P=0.003) and FFA value (HR, 3.034; 95% CI, 1.102-8.354; P=0.032) were both significantly associated with increased MALE. Elevated UA levels predicted MACE in the presence of confounders (HR, 11.446; 95% CI, 1.367-95.801 P=0.023). CONCLUSION Pre-procedure hsCRP and FFA levels could serve as predictors of adverse events after EVT in patients with arterial femoropopliteal occlusive disease. The role of UA in MACE may warrant further investigation, because the correlation is not as powerful as the other two in the study.

Kernel size, one of the traits that determine wheat yield, is controlled by multiple quantitative trait loci. Polish wheat (Triticum polonicum) has elongated and plump kernel and is a valuable material for breeding high-yielding wheat cultivars. However, genes or loci determining kernel length (KL) in Polish wheat are unknown. We identified and validated a major KL gene, KL-PW, at the P1 locus in Polish wheat. KL-PW is VRT-A2, which encodes a MIKC-type MADS-box protein (MADS55). An insertion/deletion mutation in intron 1 of VRT-A2PW led to an alternatively spliced transcript, VRT-A2PW2. Quantitative PCR analysis showed that VRT-A2PW was more highly expressed in developing seeds than was VRT-A2Ailanmai. Brassinosteroid (BR) sensitivity experiment and the expression of BR-related genes indicated that VRT-A2PW functions as a positive regulator of BR responses. VRT-A2PW significantly increased KL of wheat. These findings not only reveal the molecular basis of KL-PW in controlling KL, but also provide a valuable genetic resource for increasing kernel size in wheat.

No study on the relationship between hepatic steatosis and sex hormone levels in male patients with chronic hepatitis B (CHB) infection has been conducted.We aimed to investigate the association between serum sex hormones and hepatic steatosis among a cohort of males with CHB.In this cross-sectional study, 268 male patients with CHB were enrolled. All participants underwent anthropometric measurement, blood testing, and FibroScan test. Multiple logistic regression analysis was used to investigate the association of serum sex hormones with hepatic steatosis.We included 137 males with and 131 without hepatic steatosis in this study. Subjects with serum testosterone (T) levels in the highest tertile had an odds ratio (OR) (95% confidence interval [CI]) of 0.35 (0.18-0.70) (P for trend=0.003); those with serum prolactin (PRL) levels in the highest tertile had an OR (95%CI) of 0.21 (0.10-0.45) (P for trend0.001); and those with serum estradiol/testosterone (E2/T) in the highest tertile had an OR (95%CI) of 4.02 (1.97-8.20) (P for trend0.001) for hepatic steatosis.Lower serum total T and PRL levels and higher total E2/T are independently associated with presence of hepatic steatosis in male patients with CHB.

OBJECTIVE The dynamic adaptive immune responses elicited by the inactivated virus vaccine, CoronaVac, remain elusive. METHODS In a prospective cohort of 100 SARS-CoV-2 naïve healthcare professionals who received two doses of CoronaVac, we analyzed SARS-CoV-2-specific humoral and cellular responses at four different timepoints, including before vaccination (T1), 2 weeks after the first dose (T2), 2 weeks after the booster dose (T3), and 8-10 weeks post the booster dose (T4). SARS-CoV-2-specific antibodies, serum neutralizing activities, peripheral B cells, CD4+ and CD8+ T cells, and their memory subsets were simultaneously measured in this cohort. RESULTS SARS-CoV-2 Spike-specific IgG responses reached the peak (geometric mean titer [GMT] 54827, 30969-97065) after two doses and rapidly declined (GMT 502, 212-1190) at T4, whereas suboptimal IgA responses were detected (GMT 5, 2-9). Spike-specific circulating B cells (0.60%, 0.46-0.73% of total B cells) and memory B cells (1.18%, 0.92-1.44% of total memory B cells) were effectively induced at T3 and sustained over time (0.33%, 0.23-0.43%; 0.87%, 0.05-1.67%, respectively). SARS-CoV-2-specific circulating CD4+ T cells (0.57%, 0.47-0.66%) and CD8+ T cells (1.29%, 1.04-1.54%) were detected at T3. At T4, 0.78% (0.43-1.20%) of memory CD4+ T cells and 0.68% (0.29-1.30%) of memory CD8+ T cells were identified as SARS-CoV-2 specific, while 0.62% (0.51-0.75%) of CD4+ T cells and 0.47% (0.38-0.58%) of CD8+ T cells were SARS-CoV-2 specific terminally differentiated effector memory cells. Furthermore, age and interval between doses affected the magnitude of CoronaVac induced immune responses. SARS-CoV-2 memory CD4+ T cells was strongly associated with both RBD-specific memory B cells (r=0.87, p, Graphical abstract Image 1

Arbuscular mycorrhiza (AM) is a mutualistic symbiosis formed between most land plants and Glomeromycotina fungi. During symbiosis, plants provide organic carbon to fungi in exchange for mineral nutrients. Previous legume studies showed that the required for arbuscular mycorrhization2 (RAM2) gene is necessary for transferring lipids from plants to AM fungi (AMF) and is also likely to play a “signaling” role at the root surface. To further explore RAM2 functions in other plant lineages, in this study, two rice (Oryza sativa) genes, OsRAM2 and OsRAM2L, were identified as orthologs of legume RAM2. Examining their expression patterns during symbiosis revealed that only OsRAM2 was strongly upregulated upon AMF inoculation. CRISPR/Cas9 mutagenesis was then performed to obtain three Osram2 mutant lines (-1, -2, and -3). After inoculation by AMF Rhizophagus irregularis or Funneliformis mosseae, all of the mutant lines showed extremely low colonization rates and the rarely observed arbuscules were all defective, thus supporting a conserved “nutritional” role of RAM2 between monocot and dicot lineages. As for the signaling role, although the hyphopodia numbers formed by both AMF on Osram2 mutants were indeed reduced, their morphology showed no abnormality, with fungal hyphae invading roots successfully. Promoter activities further indicated that OsRAM2 was not expressed in epidermal cells below hyphopodia or outer cortical cells enclosing fungal hyphae but instead expressed exclusively in cortical cells containing arbuscules. Therefore, this suggested an indirect role of RAM2 rather than a direct involvement in determining the symbiosis signals at the root surface. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2022.

During an investigation of Xylariaceae from 2019 to 2020, isolates representing eight Nemania (Xylariacese) species were collected from Yunnan, Guizhou and Hainan Provinces in China. Morphological and multi-gene phylogenetic analyses, based on combined ITS, α-actin, rpb2 and β-tubulin sequences, confirmed that six of them are new to science, viz. Nemania camelliae, N. changningensis, N. cyclobalanopsina, N. feicuiensis, N. lishuicola and N. rubi; one is a new record (N. caries) for China and one is a known species (N. diffusa). Morphological descriptions and illustrations of all species are detailed. In addition, the characteristics of Nemania are summarised and prevailing contradictions in generic concepts are discussed.

Currently available therapies for hepatocellular carcinoma (HCC), with a high morbidity and high mortality, are only marginally effective and with sharp adverse side effects, which makes it compulsory to explore novel and more effective anticancer molecules. Chinese medicinal herbs exhibited prominent anticancer effects and were applied to supplement clinical cancer treatment. Here, we reported a compound, trilobolide-6-O-isobutyrate (TBB), isolated from the flowers of Wedelia trilobata with a markedly cytotoxic effect on HCC cells. We found that TBB time- and dose-dependently inhibited HCC cells' growth and colony formation in vitro. Moreover, TBB induced cell cycle arrest at the G2/M phase, mitochondrial caspase-dependent apoptosis, and suppressed migration and invasion, as well as the glycolysis of HCC cells. Mechanistically, our data indicated that TBB inhibited the STAT3 pathway activation by directly interacting with the TYR 640/657 sites of the STAT3 protein and decreasing the level of p-STAT3. TBB also regulated the expression of PCNA, Ki67, Cyclin B1, Cyclin E, Bax, Bcl2, MMP2/9, and PGK1 through the inhibition of the IL-6/STAT3 signaling pathway. Lastly, we confirmed that TBB effectively eliminated tumor growth without causing overt toxicity to healthy tissues in the xenograft tumor model. The exploration of anticancer activity and the underlying mechanism of TBB suggested its usage as a promising chemotherapeutic agent for HCC.

ACKNOWLEDGEMENTS : This study was partially supported by the European Maritime and Fisheries Fund MAR2020 project “VALOREJET: Valorização de espécies rejeitadas e de baixo valor commercial,” MAR-01.03.01-FEAMP0003, and by Fundação para a Ciência e Tecnologia through research contracts attributed to Vera Sequeira (CEECIND/02705/2017) and Ana Rita Vieira (CEECIND/01528/2017) and strategic project UID/MAR/04292/2019. The depletion and overexploitation of several fish stock demands for a valorisation of non-target and discarded species. Nonetheless, such species are often poorly studied, and information on their biological parameters must be gathered for effective population management. For 1 year, the reproductive strategy of the piper gurnard Trigla lyra and the red gurnard Chelidonichthys cuculus was studied by monthly samples obtained from commercial boats operating on western Portuguese coast. Both species showed a biased sex ratio towards females, especially for larger length classes. Length at first maturity could be estimated only for red gurnard (22.1 and 19.9 cm for females and males, respectively) because all piper gurnard individuals caught were mature. Piper gurnard showed determinate fecundity and a short spawning season, from November to February with a peak in January, whereas red gurnard showed indeterminate fecundity and a wide spawning season, from late December to May. The relative annual fecundity estimated for red gurnard (1893 ± 728 oocytes x g-1 eviscerated weight [EW]) was higher than the one estimated for piper gurnard (1018 ± 250 oocytes x g-1 EW). Although important information for understanding the species dynamics is presented in this study, additional information on other life-history parameters and of species landings is required. info:eu-repo/semantics/publishedVersion