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Camellia japonica, an evergreen ornamental plant in the Theaceae family, has a natural range that is now shrinking. This is evident by the fact that the species is on the verge of extinction in Laoshan Mountain (Qingdao), the northernmost area of China known to have a natural population of C. japonica. Little is known about the genetic diversity and relationships among cultivated and wild C. japonica populations. One hundred and eighty samples of six C. japonica populations were tested for genetic diversity with simple sequence repeat (SSR) markers; these included three cultivated populations, two natural populations in Qingdao, and one natural population in Daqingshan. The average values of polymorphism information content (PIC), expected heterozygosity (He), and Shannon’s information index (I) were 0.5849, 0.6385 and 1.3170, respectively, indicating that C. japonica has a high genetic diversity. The genetic diversities of the six populations in rank order were as follows: Daqingshan > Zhongshan Park > Changmenyan Island > Daguan Island > Botanical Garden > May Fourth Square. The geographical isolation of the islands had no significant influence on the genetic diversity of C. japonica. Clustering results showed that the six C. japonica populations could be grouped into three categories, and most populations were clustered according to their geographical origin and genetic background. These results also reconfirmed that the C. japonica (Naidong) population in Qingdao originated from Changmenyan Island. Genetic variation was highest within populations (89%), indicating that C. japonica can be protected at the population level. These findings will prove useful for the genetic analysis, protection, and horticultural use of C. japonica.

Aims Nitrogen (N) supply and precipitation pattern (amount and frequency) both affect plant growth. However, N deposition is increasing and precipitation regimes are changing in the context of global change. An experiment was conducted to determine the effect of precipitation pattern and N supplies on the growth of a widely distributed and planted N2-fixing alien species Robinia pseudoacacia. Methods Seedlings were grown in a glasshouse at four different N levels combined with different precipitation regimes, including three precipitation amounts, and two precipitation frequencies. After treatment for 75 days, plant height, biomass allocation, leaf and soil nutrient concentrations were measured. Results Plants under high precipitation frequency had greater biomass compared with plants lower precipitation frequency with the same amount of precipitation. Higher N supply decreased biomass allocation to nodules. Nodule growth and N2 fixation of R. pseudoacacia with low precipitation amount was more inhibited by high N deposition compared with plants received higher precipitation amount. Slightly N deposition under higher precipitation inhibited N2 fixation but did not fulfil the N need in plants. Conclusions Nitrogen deposition might inhibit N2 fixation of plants even in low level but low nitrogen in soil cannot fulfil the N need of plants, and caused N2 fixation limitation in plants during seedling stage. There was likely a shift from acquiring N mainly from N2 fixation to acquisition of N directly with root when N supply was increased. High N deposition level, and increased precipitation frequency might increase the invasion risk of R. pseudoacacia.

Introduction Rapamycin (Rapa) is an immunosuppressive macrolide that inhibits the mechanistic target of rapamycin (mTOR) activity. Thanks to its anti-proliferative effects towards different cell types, including keratinocytes and T cells, Rapa shows promise in the treatment of skin diseases characterized by cell hyperproliferation. However, Rapa skin penetration is limited due to its lipophilic nature (log P = 4.3) and high molecular weight (MW = 914 g/mol). In previous studies, new microenvironment-sensitive core multishell (CMS) nanocarriers capable of sensing the redox state of inflamed skin were developed as more efficient and selective vehicles for macrolide delivery to inflamed skin. Methods In this study, we tested such redox-sensitive CMS nanocarriers using an inflammatory skin model based on human skin explants co-cultured with Jurkat T cells. Serine protease (SP) was applied on skin surface to induce skin barrier impairment and oxidative stress, whereas phytohaemagglutinin (PHA), IL-17A, and IL-22 were used to activate Jurkat cells. Activation markers, such as CD45 and CD69, phosphorylated ribosomal protein S6 (pRP-S6), and IL-2 release were monitored in activated T cells, whereas pro-inflammatory cytokines were measured in skin extracts and culture medium. Results We found that alteration of skin barrier proteins corneodesmosin (CDSN), occludin (Occl), and zonula occludens-1 (ZO-1) as well as oxidation-induced decrease of free thiol groups occurred upon SP-treatment. All Rapa formulations exerted inhibitory effects on T cells after penetration across ex vivo skin. No effects on skin inflammatory markers were detected. The superiority of the oxidative-sensitive CMS nanocarriers over the other formulations was observed with regard to drug delivery as well as downregulation of IL-2 release. Conclusion Overall, our results demonstrate that nanocarriers addressing features of diseased skin are promising approaches to improve the topical delivery of macrolide drugs., Graphical Abstract

Phragmites australis is the dominant species in the Yellow River Delta and plays an important role in wetland ecosystems. In order to evaluate the relationship between phenotypic variation and environmental factors, explore how functional traits respond to changes in electrical conductivity and soil water content, and reveal the ecological strategies of P. australis, we investigated the ecological responses of P. australis to soil properties based on 96 plots along the coastal–inland regions in the Yellow River Delta of China. Within the range of soil water content (SWC, 9.39%–36.92%) and electrical conductivity (EC, 0.14–13.29 ms/cm), the results showed that (a) the effects of salinity were more important than the soil water content for the characterization of the morphological traits and that plant functional traits including leaf traits and stem traits responded more strongly to soil salinity than soil water content; (b) compared with morphological traits such as average height and internode number, physiological traits such as SPAD value, as well as morphological traits closely related to physiological traits such as specific leaf area and leaf thickness, showed stronger stability in response to soil water and salinity; and (c) under the condition of high electrical conductivity, P. australis improved its water acquisition ability by increasing indicators such as leaf water content and leaf thickness. In addition, with the increase in plant tolerance to stress, more resources were used to resist external stress, and the survival strategy was inclined toward the stress tolerator (S) strategy. Under low EC conditions, P. australis increased specific leaf area and leaf area for its growth in order to obtain resources rapidly, while its survival strategy gradually moved toward the competitor (C) strategy., In the Yellow River Delta, soil salinity was the main factor that influenced the functional traits of P. australis; the physiological traits are more stable than morphological traits in response to environmental variables; and competition and stress tolerator were the main strategies of P. australis.

The formation and development of venous thromboembolism accounts for considerable morbidity and mortality. One of the most common methods to treat thromboembolism is the use of thrombolytic drugs to activate fibrinolytic protease. The aim of this treatment was to initiate the lysis of fibrin; however, there are many side-effects associated with this form of treatment. Herein, we fabricated chiral Co 3 O 4 supraparticles (SPs) with a g -factor of up to 0.02 at 550 nm and paramagnetic performance applied in the treatment of thromboembolism under an electromagnetic field (MF). In vitro experiments showed that d -SPs degraded blood clot within 8 hours under MF. Compared to l -SPs, d -SPs exhibited much stronger thrombolytic ability and effectively enhanced the survival rate of thrombosis model mice more than 70% in the 25 d of observation. The results of mechanism study showed that under MF, the level of reactive oxygen species (ROS) produced by d -SPs were 1.5 times higher than that of l -SPs, which might be attributed to the chiral-induced spin selectivity effects. This discovery paves an inspired pathway to develop chiral nanomaterials for the efficient treatment of thrombosis under MF.

Soil legacies mediate interactions between native and introduced plants, contributing to both invasion and biotic resistance to invasion. Given that nitrogen deposition can promote allelochemical release, reduce the benefits of soil microbes, and affect trait plasticity, nitrogen deposition likely alters soil legacies as well. However, it is not clear how mechanisms that facilitate adaptation to soil legacies are altered by nitrogen deposition. In a greenhouse setting, we investigated how an invasive and a native plant in northern China (Rhus typhina and Ailanthus altissima, respectively) acclimate to soil legacies and how these dynamics change with nitrogen availability. We measured plant functional traits, soil microbial abundance, microbial enzyme activities, and soil allelopathic effects to characterize plant responses to soil legacies from plants of the same and of the other species. Rhus typhina had a stronger growth response to soil legacies than did A. altissima. Rhus typhina established a novel plant-soil feedback by increasing fungi and bacteria, changing the composition of the microbial community, and effectively transforming negative effects of soil allelopathy to positive effects. Nitrogen deposition promoted the growth of R. typhina and alleviated the negative effects of heterospecific soil legacies on the performance of R. typhina. Invasive plants can acclimate to the soil legacies of native species through a combination of high trait plasticity, manipulating soil microbes, and establishing novel plant-soil feedbacks. Nitrogen deposition can facilitate invasive species acclimating to soil legacies by monopolizing nitrogen absorption, though this may diminish the benefit of soil microbes.

A high-throughput screening system was established by employing enhanced green fluorescent protein as a screenable fusion tag to evaluate the expression and secretion of a lytic polysaccharide monooxygenase (MtC1LPMO) using 20 Sec-type signal peptides (SPs) from Bacillus amyloliquefaciens 111018. Among these, 10 SPs were found to be better than the native SP of MtC1LPMO. The protein expression and secretion levels using SP12 (MNITNWAAILQLQSMALQSISNTGTASS) were the highest among all SPs, with 4.1- and 2.1-fold increases over the native SP, respectively. Then, the amino acids of the 10 best SPs were analyzed, and the results indicated that the most abundant amino acid of the N-region was K, those of the H-region were L, F, A and V, and the C-region contained an AXA motif. Additionally, we found that the protein expression level gradually improved along with the increasing folding free energies of the SP-encoding part of the mRNA. Finally, the SPs were rationally designed to improve the expression and secretion level of MtC1LPMO. An increased positive charge of the SP N-region was found to enhance the protein expression and secretion level, as long as the folding free energy of the mRNA did not change significantly.

Due to their low nitrogen utilization efficiency (NUtE) oilseed rape (OSR) crops require large quantities of nitrogen (N) fertilizer to achieve acceptable economic yields. To dissect the underlying mechanisms of oilseed NUtE, we investigated the relationships between NUtE, N uptake, N remobilization and leaf senescence by growing three N-efficient (genotypes with high NUtE at deficient N supplies) and three N-inefficient (ones with low NUtE at deficient N supplies) genotypes in pot experiments. These genotypes showed similar plant N uptake before flowering and similar post-anthesis N remobilization from leaf and stem, but after flowering N-efficient genotypes exhibited higher root N uptake than N-inefficient genotypes. Besides, the pod biomass and leaf N concentration of N-efficient genotypes were higher than those of N-inefficient genotypes at 50 days after flowering, and we observed delayed leaf senescence in N-efficient genotypes. Correspondingly, gene transcripts involved in photosynthesis and carbon metabolism were abundant in green mature leaves of N-efficient genotype, while the expression of genes involved in ubiquitin mediated proteolysis pathway were more abundant in N-inefficient mature leaves. The stay-green phenotype of the N-efficient OSR after flowering provides sufficient photosynthate for root N uptake activity and pod development, and in turn, the prolonged post-anthesis N uptake provides more N to pods, leading to less leaf N remobilization requirement. This work has identified traits important for breeding OSR cultivars with improved NUtE.

Preliminary studies observed a lower growth activity during the vegetative stage with higher growth attributes at the pod-filling stage among the high nitrogen (N) utilisation efficiency (NUtE) oilseed rape (Brassica napus L.) genotypes, compared with the low NUtE genotypes. Therefore, we hypothesised that there would exist a critical growth stage when distinctive phenotypic traits are exhibited to regulate yield formation and NUE. A field experiment and a hydroponic culture were conducted to characterise the differences in shoot and root physiological indicators of the high and low NUtE oilseed rape genotypes at seedling, bud, bolting, flowering and pod-filling stages. We found that flowering was the critical period when the reverse growth habit occurred between high and low NUtE genotypes. The high NUtE genotypes displayed larger values of root traits, stronger N uptake kinetics parameters, higher activity of leaf glutamine synthetase (GS) and glutamate synthetase (GOGAT), larger SPAD values and net photosynthetic rate, ultimately leading to higher seed yield and NUE. Our results indicate that flowering is the critical growth stage to distinguish the high from low NUtE oilseed rape genotypes, and plant breeders may focus on selecting root and shoot phenotypic traits from flowering stage onwards to achieve both high yields and NUE for oilseed rape genotypes.

Myosins are ATP-dependent actin-based molecular motors critical for diverse cellular processes like intracellular trafficking, cell motility, and cell invasion. During cell division, myosin MYO10 is important for proper mitotic spindle assembly, the anchoring of the spindle to the cortex, and positioning of the spindle to the cell mid-plane. However, myosins are regulated by myosin regulatory light chains (RLCs), and whether RLCs are important for cell division has remained unexplored. Here, we have determined that the previously uncharacterized myosin RLC Myl5 associates with the mitotic spindle and is required for cell division. We show that Myl5 localizes to the leading edge and filopodia during interphase and to mitotic spindle poles and spindle microtubules during early mitosis. Importantly, depletion of Myl5 led to defects in mitotic spindle assembly, chromosome congression, and chromosome segregation and to a slower transition through mitosis. Furthermore, Myl5 bound to MYO10 in vitro and co-localized with MYO10 at the spindle poles. These results suggest that Myl5 is important for cell division and that it may be performing its function through MYO10.

Increasing post-flowering nitrogen (N) uptake is likely to improve seed N accumulation, ultimately leading to greater seed yield and N use efficiency (NUE). A comprehensive study on contrasting N utilization efficiency (NUtE) winter oilseed rape genotypes was conducted for three years (2017–2020) to unravel the effect of post-flowering N uptake on seed N accumulation. Compared to the low NUtE genotype, the high NUtE genotype displayed respectively 31%, 28% and 70% greater root biomass, length and volume, along with 40%, 44%, 46% and 82% higher root nitrate reductase (NR), glutamine synthetase (GS), glutamate synthetase (GOGAT), and glutamate dehydrogenase (GDH) activities after flowering. These were accompanied by a significant increase (P < 0.05) in catalases (134%), peroxidase (45%), glutathione (45%) and ascorbate peroxidase (41%), leading to 27% higher seed N accumulation, 23% higher seed yield and 60% higher NUE. Stronger post-flowering N uptake potential for high NUtE genotype was a main contributor to the enhanced seed N accumulation, and ultimately increased seed yield and NUE.

Failed neuroprotection leads to the initiation of several diseases. SOX1 plays many roles in embryogenesis, oncogenesis, and male sex determination, and can promote glioma stem cell proliferation, invasion, and migration due to its high expression in glioblastoma cells. The functional versatility of the SOX1 gene in malignancy, epilepsy, and Parkinson's disease, as well as its adverse effects on dopaminergic neurons, makes it an interesting research focus. Hence, we collate the most important discoveries relating to the neuroprotective effects of SOX1 in brain cancer and propose hypothesis worthy of SOX1's role in the survival of senescent neuronal cells, its roles in fibroblast cell proliferation, and cell fat for neuroprotection, and the discharge of electrical impulses for homeostasis. Increase in electrical impulses transmitted by senescent cells affects the synthesis of neurotransmitters, which will modify the brain cell metabolism and microenvironment.

In this study, Ougan juice (OJ) and lactic acid bacteria fermented Ougan juice (FOJ) were investigated individually for their capability of preventing obesity in high-fat diet (HFD)-fed C57BL/6J mice. After being administered with OJ or FOJ for 10 weeks, the body weight gain, hyperlipidemia, and gut microbiota dysbiosis of HFD-fed mice were examined. The results showed that OJ or FOJ supplementation inhibited weight gain, lowered fat accumulation, reduced liver steatosis, improved glucose homeostasis and insulin sensitivity, increased brown adipose tissue (BAT) activity, and promoted white adipose tissue (WAT) browning. Both OJ and FOJ additions increased the diversity of gut microbiota. OJ reduced the relative abundance of phylum Erysipelatoclostridiaceae and genus Erysipelatoclostridium and remarkably increased SCFA-producing bacteria Blautia, while FOJ reduced the ratio of Firmicutes to Bacteroidetes and enhanced the relative abundance of family Lactobacillaceae. Spearman's correlation analysis revealed that Akkermansia, Dubosiella, and Muribaculaceae were significantly negatively correlated with obesity-related indexes. In general, FOJ exhibited a better inhibitory effect on obesity than OJ, and the possible inhibitory mechanism lies in promoting WAT browning and increasing intestinal probiotics. This study provides the guidance for developing fermented Ougan juice as an obesity-inhibiting functional food.

Graphical abstract, Introduction Eugenol is a major component of essential oils of several plants, it exhibits significant antiparasitic and acaricidal activities, yet its molecular targets remain unknown. Objectives We aimed to systematically investigate the mechanism of action and the potential targets of eugenol against P. cuniculi, and evaluate the safety for laying the theoretical foundation for clinical application as an acaricide. Methods Using RNA-Seq analysis, surface plasmon resonance analysis and RNA interference assay, the mode of action of eugenol against Psoroptes cuniculi was investigated. The effect on the mitochondrial membrane potential and complex I of PC12 cells and C6/36 cells was assayed to investigate the species specificity of eugenol in insects and mammals. Finally, a safety evaluation of eugenol in vivo was performed. Results Eugenol inhibited complex I activity of the mitochondrial respiratory chain in the oxidative phosphorylation pathway by binding to NADH dehydrogenase chain 2 and resulted in the death of mites. The inhibition rates were 37.89% for 50 μg/mL and 60.26% for 100 μg/mL, respectively. Further experiments indicated that the difference in the complex I sequence between insects and mammals led to the different affinity of eugenol to specific peptide, resulting in species specificity. Eugenol exhibited significant inhibitory effects against the mitochondrial membrane potential and complex I in Aedes albopictus C6/36 cells but was not active in rat PC12 cells. Insect cells were particularly sensitive to eugenol. In contrast to the known inhibitor rotenone, eugenol had better safety and did not result in Parkinson’s disease or other diseases in rats. Conclusion This is the first report on acaricidal eugenol targeting complex I of the mitochondrial respiratory chain. This work lays the foundation for the development of eugenol as an environmentally alternative acaricidal agent.

BACKGROUND: Due to adipose-derived stem cells (ASCs) being easy to obtain, their rapid proliferation rate, and their multidirectional differentiation capabilities, they have been widely used in the field of regenerative medicine. With the progress of decellularized adipose tissue (DAT) and adipose tissue engineering research, the role of DAT in promoting angiogenesis has gradually been emphasized. METHODS: We examined the biological characteristics and biosafety of DAT and evaluated the stem cell maintenance ability and promotion of growth factor secretion through conducting in vitro and in vivo studies. RESULTS: The tested ASCs showed high rat:es of proliferation and adhered well to DAT. The expression levels of essential genes for cell stem maintenance, including OCT4, SOX2, and Nanog were low at 2–24 h and much higher at 48 and 96 h. The Adipogenic expression level of markers for ASCs proliferation including PPARγ, C/EPBα, and LPL increased from 2 to 96 h. Co-culture of ASCs and DAT increased the secretion of local growth factors, such as VEGF, PDGF-bb, bFGF, HGF, EGF, and FDGF-bb, and secretion gradually increased from 0 to 48 h. A model of full-thickness skin defects on the back of nude mice was established, and the co-culture of ASCs and DAT showed the best in vivo treatment effect. CONCLUSION: The application of DAT promotes wound healing, and DAT combined with ASCs may be a promising material in adipose tissue engineering and regenerative medicine.

Polyploidization and hybridization are very common in natural plant species, and mixed-ploidy species provide a unique opportunity to study the effects of evolutionary history, local abundance, and ploidy level on the direction and extent of introgression between intraspecific lineages. First, we delimited two morphologically cryptic lineages of Phragmites australis Trin. ex Steud. in temperate China using 11 nuclear microsatellites and two chloroplast DNA fragments with 225 samples from China as well as 11 samples from Oceania and Europe. Our evidence supported that haplotype O and haplotype P were two relatively independent lineages with low and high ploidy levels, respectively; haplotype M might be ancient and could have undergone a complex evolutionary history. Then we examined the lineage divergence and compared the introgression patterns between two major lineages along geographical and abundance gradients with a large number of samples (n = 1067) collected from China. The sympatric coexistence of two lineages in north and northeast China implies an ongoing or potential introgression between them. Cline analysis showed that the level of genetic admixture were significantly correlated with longitude rather than latitude. Our results also suggested that ploidy level could deeply influence the introgression asymmetry, and the effect of the current local abundance on introgression might be covered by the past coexistence time driven by phylogeographic history. Our study draws a baseline for future research on the ecological and evolutionary consequences of migration and introgression of Chinese P. australis under global change.

Microbial processes are being developed to transform flavonoid glycosides to varieties of metabolites with higher bioavailability. The aim of this study was to determine the metabolic activity and survival of five lactic acid bacteria (LAB) stains (L. rhamnosus LRa05, L. casei LC89, L. plantarum N13, L. acidophilus LA85, and L. brevis LB01) in two different citrus flavanone standards (hesperetin-7-O-rutinoside and naringenin-7-O-rutinoside). The enzymatic activity, metabolites, antioxidant activities, and α-glucosidase inhibition property in the two standards were also investigated before and after incubated with LAB. The medium contained standards permitted survival of the five LAB stains. All strains exhibited β-glucosidase activity. Of the five LAB strains tested, just L. plantarum N13 and L. brevis LB01 have the ability to metabolize hesperetin-7-O-rutinoside, only L. plantarum N13, L. acidophilus LA85, and L. brevis LB01 could metabolize naringenin-7-O-rutinoside, moreover, L. acidophilus LA85l was the strain with the highest biotransformation ratio of naringenin-7-O-rutinoside. L. acidophilus LA85 and L. plantarum N13 can degrade naringenin-7-O-rutinoside into naringenin. L. brevis LB01 can degrade hesperetin-7-O-rutinoside into hesperetin, 3-(4'-hydroxyphenyl)-2-propenoic acid, 3-(3'-hydroxy-4'-methoxyphenyl)hydracrylic acid, and 3-(4'-hydroxyphenyl)propionic acid. Incubation of L. acidophilus LA85 in naringenin-7-O-rutinoside solution supposed no apparent influence in the biological activities that tested. L. acidophilus LA85 may potentially contribute to the bioavailability of citrus flavanones, and to be applied as functional cultures to obtain more bioavailable and bioactive metabolites in food products or in the human gastrointestinal tract.

Isoquinoline alkaloids, an important class of N-based heterocyclic compounds, have attracted considerable attention from researchers worldwide since the early 19(th) century. Over the past 200 years, many compounds from this class were isolated, and most of them and their analogs possess various bioactivities. In this review, we survey the updated literature on bioactive alkaloids and highlight research achievements of this alkaloid class during the period of 2014–2018. We reviewed over 400 molecules with a broad range of bioactivities, including antitumor, antidiabetic and its complications, antibacterial, antifungal, antiviral, antiparasitic, insecticidal, anti-inflammatory, antioxidant, neuroprotective, and other activities. This review should provide new indications or directions for the discovery of new and better drugs from the original naturally occurring isoquinoline alkaloids.

Potato late blight, caused by the oomycete pathogen, Phytophthora infestans de Bary, is one of the most devastating plant diseases in the world. It is of great importance to screen disease resistance germplasms and elucidate the molecular mechanisms underlying the resistance. In a previous study, Huasong 7, a tetraploid potato variety, was identified to possess resistance to multiple P. infestans races at various levels. In the present study, Huasong 7 was inoculated with an incompatible isolate 428–2 and a compatible P. infestans isolate T30–4, and gene expression was profiled at 0, 24, 48, and 72 h post inoculation. The results showed that more gene expression differentiation was observed in the compatible interaction induced by T30–4 than that in the incompatible interaction induced by 428–2. The differentially expressed genes induced by 428–2 were significantly enriched in GO terms associated with immune responses and the regulation of defense responses, as well as KEGG pathways related to antioxidant activities, for example, the glutathione metabolism. In contrast, differentially expressed genes induced by T30–4 were significantly enriched in GO terms associated with transcription, translation, and biosynthesis of macromolecules, as well as disease resistance related KEGG pathways, such as phosphoinositide metabolism, phospholipid signaling system, and receptor-mediated endocytosis. Our findings indicated that Huasong 7 responds to incompatible P. infestans isolate through disease resistance and host defense related pathways; while it responds to compatible isolate T30–4 with basic responses. It is also possible that T30–4 contains other unknown Avr genes. In summary, our study could provide an important reference for a better understanding of potato late blight resistance mechanisms of both compatible and incompatible interactions.

The product productivity of Chinese hamster ovary (CHO) cells need to be further increased and solved. Promoter and selection marker are essential elements for transgene expression. We investigated the effects of promoters, Woodchuck hepatitis post-transcriptional regulatory (WPRE) element, and selection marker on transgene expression in CHO cells. A weakened form of resistance gene neomycin phosphotransferase (NPT) and WPRE element were cloned into the expression vectors driven by the cytomegalovirus (CMV) and simian virus 40 (SV40) promoters, following by transfecting into CHO cells and screened under G418 pressure. qRT-PCR and Flow cytometry were used to detect the expression level of eGFP gene, and transgene copy number was determined using qPCR technology. Western blot and ELISA were utilized to analyze the expression levels of cetuximab. The weakened NPT gene enhanced eGFP expression both under the CMV or SV40 promoters. Moreover, WPRE, combined with the NPT gene considerably enhanced eGFP expression in CHO cells. Furthermore, the CMV promoter and WPRE combination resulted in higher cetuximab expression levels than CMV promoter alone. However, WPRE could not enhance the transgene expression under the circumstance of CMV promoter and weakened NPT gene combination, suggesting that the effects of the WPRE combined with selection markers were specificity.

Tamarix chinensis is a dominant species in both the Yellow River Delta Natural Reserve and Changyi National Marine Specific Protection Area, Laizhou Bay, China, and plays an important role in wetland ecosystems. We investigated the relationship between T. chinensis and environmental factors, and the influences of T. chinensis on understory plants. We selected four T. chinensis communities with different hydrologic conditions and salinities in the Yellow River Delta Natural Reserve and the Changyi National Marine Specific Protection Area. The results showed that the growth of T. chinensis was negatively correlated with soil salinity. Generally no fertile island effects or salt island effects of T. chinensis on herbaceous plant communities were found in the coastal area of Laizhou Bay, China. In areas with high salinity, T. chinensis provided nursery effects for understory herbaceous species, increasing species diversity and enhancing plant growth. In low-salinity environments, T. chinensis appeared to restrain the growth of understory plants. This study observed different influences of T. chinensis on herbaceous species between the Yellow River Delta Natural Reserve and Changyi National Marine Specific Protection Area, providing scientific basis and theoretical foundation for investigating plant distribution and ecological community succession in Laizhou Bay, China.

Objective To explore whether auricular concha eletroacupuncture (ACEA) is effective in regulating lipid-lipoprotein metabolism in rats submitted to cold stress. Methods Thirty-six adult male Sprague–Dawley rats were placed in four groups(9 rats in each group), the rats in three groups of which were submitted to cold stress for fourteen days, the last one of which was a control group. After the cold stress process, in those three groups, the rats of one group were with no treatment (stress only), two were treated with either ACEA or auricular margin eletroacupuncture (AMEA) repeated for fourteen days. On the 14th day, all the rats were sacrificed after all experimental procedure for blood sampling. Blood glucose, triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were tested by using the collected serum. Plasma lecithin-cholesterol acyltransferase (LCAT) was measured with ELISA kit. Results ACEA down-regulated the TG level (P Conclusion ACEA played an important role in regulating lipid-lipoprotein metabolism in rats submitted to cold stress.

Widespread plant species generally have high intraspecific variation in functional traits, which is reflected in their great variety of phenotypes. This variety can result from both genetic differences due to local adaptation and phenotypic plasticity. With high intraspecific variation and nearly global distribution, the common reed Phragmites australis is a suitable model species for studying the underlying mechanisms of intraspecific trait variation. In this study, 71 genotypes of P. australis from seven phylogeographic groups were transplanted into two replicate common gardens located in very different climates: northern Europe and mid-east Asia. We measured seven functional traits of all these genotypes over the growing season, including shoot height, maximum biomass per shoot, shoot density, node number per stem, leaf life span, flowering occurrence and flowering date. Our aim was to assess the relative effects of genetic (phylogeographic origin) and environmental (common garden) status, and interactions between them, on intraspecific variation in functional traits of P. australis. We found common garden having the strongest influence on most functional traits studied. All traits except flowering occurrence varied significantly across gardens, revealing the important role of phenotypic plasticity on trait variation in P. australis. We also found significant differences in trait variation among the different phylogeographic groups of P. australis and, thus, evidence for genetically determined intraspecific variation in the morphological and life-history traits addressed in this study. All functional traits showed significant (p ≤ 0.0054), albeit minor to moderately explained (R2 ≤ 0.57), latitudinal patterns in both gardens. Covariation of multiple traits was similar in the two gardens. Phenotypic plasticity was trait- specific, and the plasticity of shoot height and maximum biomass per shoot increased towards higher latitude of genotypic origin. Our results indicate that the latitude of origin affects the evolution of functional traits, as well as their phenotypic plasticity. Since phenotypic plasticity is a crucial mechanism for acclimation and evolution, our findings support the role of gene-based adaptive phenotypic plasticity in plant evolution. The intraspecific spatial variation in functional traits and their phenotypic plasticity can help predict species distribution, persistence and invasion under global climate change.

Human tissue plasminogen activator was the first recombinant therapy protein that successfully produced in Chinese hamster ovary cells in 1986 and approved for clinical use. Since then, more and more therapeutic proteins are being manufactured in mammalian cells, and the technologies for recombinant protein production in this expression system have developed rapidly, with the optimization of both upstream and downstream processes. One of the most promising strategies is expression vector cassette optimization based on the expression vector cassette. In this review paper, these approaches and developments are summarized, and the future strategy on the utilizing of expression cassettes for the production of recombinant therapeutic proteins in mammalian cells is discussed.

Coronavirus Disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) is continuously and rapidly circulating at present. Asymptomatic patients have been proven to be contagious and thus pose a significant infection control challenge. Here we describe the characteristics of asymptomatic patients with SARS-CoV-2 infection in Jinan, Shandong province, China. A total of 47 patients with confirmed COVID-19 were recruited. Among them, 11 patients were categorized as asymptomatic cases. We found that the asymptomatic patients in Jinan were relatively young and were mainly clustered cases. The laboratory indicators and lung lesion on chest CT were mild. No special factors were found accounting for the presence or absence of symptoms. The presence of asymptomatic patients increased the difficulty of screening. It is necessary to strengthen the identification of such patients in the future.

Weeds and insect pests are two important biotic stresses resulting in yield loss in rice, and wide compatibility is the essential characteristic of breeding inter-subspecific hybrid rice. This study focused on glyphosate resistance, lepidopteran resistance and wide compatibility as well as identification of molecular and some agronomic characteristics of transgenic male sterile line E1C4008S. The results indicated that glyphosate resistance gene Epsps# and lepidopteran resistance gene Cry1ca# were transferred into japonica wide compatibility male sterile line 4008S by Agrobacterium-mediated method, and four independent transformation events named E1C4008S-1, E1C4008S-2, E1C4008S-3 and E1C4008S-4 were obtained, in which E1C4008S-3 and E1C4008S-4 were of single copy insertion. The EPSPS protein contents of E1C4008S-3 and E1C4008S-4 in different organs were significantly different both in descending order of leaf > stem > root, and ranged from 300.58 to 1410.69 μg/g at the tillering stage. The glyphosate tolerable concentration (dosage) of E1C4008S-3 and E1C4008S-4 reached at least 16 g/L (54.42 kg/hm2), and the seeds of E1C4008S-4 can germinate normally on the medium containing 1 g/L glyphosate. The CRY1C protein contents of E1C4008S-3 and E1C4008S-4 in different organs were significantly different both in descending order of leaf > stem > root, and ranged from 0.62 to 2.43 μg/g at the tillering stage. The larvae mortalities of rice leaf rollers fed on leaves of E1C4008S-3 and E1C4008S-4 for 5 d were 95.35% and 97.77%, respectively, while the average mortalities of silkworms fed with protein extracts from leaves of E1C4008S-3 and E1C4008S-4 reached 94.55% and 83.64%, respectively. The results suggested that wide compatibility and evaluated agronomic traits of E1C4008S-4 were not significantly changed by insertion of the exogenous genes. Overall, a novel male sterile germplasm with glyphosate resistance, lepidopteran resistance and wide compatibility was verified to be developed in rice. Keywords: rice, glyphosate resistance, lepidopteran resistance, wide compatibility, male sterile line, bioengineered crop

Phytohormonal interactions are crucial for plant development. Auxin and cytokinin (CK) both play critical roles in regulating plant growth and development; however, the interaction between these two phytohormones is complex and not fully understood. Here, we isolated a wild apple (Malus sieversii Roem) GRETCHEN HAGEN3 (GH3) gene, MsGH3.5, encoding an indole-3-acetic acid (IAA)-amido synthetase. Overexpression of MsGH3.5 significantly reduced the free IAA content and increased the content of some IAA-amino acid conjugates, and MsGH3.5-overexpressing lines were dwarfed and produced fewer adventitious roots (ARs) than the control. This phenotype is consistent with the role of GH3 in conjugating excess free active IAA to amino acids in auxin homeostasis. Surprisingly, overexpression of MsGH3.5 significantly increased CK concentrations in the whole plant, and altered the expression of genes involved in CK biosynthesis, metabolism and signaling. Furthermore, exogenous CK application induced MsGH3.5 expression through the activity of the CK type-B response regulator, MsRR1a, which mediates the CK primary response. MsRR1a activated MsGH3.5 expression by directly binding to its promoter, linking auxin and CK signaling. Plants overexpressing MsRR1a also displayed fewer ARs, in agreement with the regulation of MsGH3.5 expression by MsRR1a. Taken together, we reveal that MsGH3.5 affects apple growth and development by modulating auxin and CK levels and signaling pathways. These findings provide insight into the interaction between the auxin and CK pathways, and might have substantial implications for efforts to improve apple architecture.

Misfolding and accumulation of amyloidogenic proteins into various forms of aggregated intermediates and insoluble amyloid fibrils is associated with more than 50 human diseases. Large amounts of high-quality amyloid proteins are required for better probing of their aggregation and neurotoxicity. Due to their intrinsic hydrophobicity, it is a challenge to obtain amyloid proteins with high yield and purity, and they have attracted the attention of researchers from all over the world. The rapid development of bioengineering technology provides technical support for obtaining large amounts of recombinant amyloidogenic proteins. This review discusses the available expression and purification methods for three amyloid proteins including amyloid β-protein, tau, and α-synuclein in microbial expression systems, especially Escherichia coli, and discusses the advantages and disadvantages of these methods. Importantly, these protocols can also be referred to for the expression and purification of other hydrophobic proteins.

Salicylic acid (SA) is an important signaling substance that plays an important role in plant growth, development and disease resistance. In order to further understand the role of the SA pathway in potato disease resistance and identify SA signaling key genes, gene expression profiling of the late blight resistance genotype SD20 was performed under exogenous SA application. A total of 28 572 unigenes were assembled, of which 4 564 were differentially expressed. Analysis of differentially expressed genes (DEGs) showed that multiple signaling pathways such as SA, jasmonic acid, ethylene, abscisic acid, auxin, and brassinolide were involved in response to exogenous SA. Many plant defense signal-related genes involved in protein serine/threonine kinase activity and plant-pathogen interaction, were significantly enriched. These were consistent with the interaction results of SD20 and Phytophthora infestans in our previous study, indicating that exogenous SA stimulated the resistance response and initiated a similar defense pathway compared to pathogen infection in SD20, which confirmed crosstalk of the SA signaling pathway with a pathogen-induced disease resistance signal pathway in plant. Moreover, transcriptome analysis revealed that ROS1 was positively regulated by SA in potato for the first time. SA-induced gene expression profiling provides insight into SA signaling and its mechanisms in disease defense systems. Keywords: potato, salicylic acid, transcriptome, pathway, defense response

Objectives: Lung adenocarcinomas with or without epidermal growth factor receptor (EGFR) mutations have shown different drug effects against EGFR inhibitors. But it is not very clear if EGFR mutation status affects the biological behavior of lung adenocarcinoma, because tumor gene regulation is very complicated and can be affected by many factors. We aimed to explore if EGFR mutation status is related with tumor malignant degree by investigating the relevance of EGFR mutation status with DNA content and aneuploid peaks of lung adenocarcinoma cells in pleural fluids without using EGFR-TKIs. Materials and Methods: 591 cases of lung adenocarcinoma patients in Hebei Tumor Hospital who had undergone EGFR gene detection and DNA quantitative analysis were collected from January 2012 to August 2018.They were divided into two groups: EGFR mutant group and non-mutant group. EGFR mutations were detected by Amplification Refractory Mutation System (ARMS) and ABI 7500 Fluorescence quantitative PCR with pleural effusions. DNA content and aneuploid peaks were detected by LD DNA image cytometry (DNA-ICM). Rank-sum test of SPSS 16 was used for statistical analysis. Results: The maximum DI, the mean DI of the first 20 cells greater than 5C, the percentage of cells greater than 5C and the number of cells greater than 9C of the first 20 cells in the mutant group were all higher than those in the non-mutant group, having statistical significance (p

Sugar alcohols, also known as polyols, have been widely applied in food products for their unique properties. In this work, the effect of soaking treatments composed of the sugar alcohols xylitol, sorbitol, erythritol, lactitol, mannitol, maltitol, and isomaltitol on the quality properties of peeled Pacific white shrimp during frozen storage were evaluated. Samples treated with water and sodium pyrophosphate (Na4P2O7) were designated as the negative and positive controls, respectively. The results indicated that significant improvements were found in thawing, pressing, and cooking loss, myofibrillar protein content, myofibrillar Ca2+-ATPase activity, and textural variables in xylitol- and mannitol-treated shrimp as compared to that of control samples. Water mobility analyses showed that xylitol and mannitol effectively increased the relaxation times T21 and T22 and the percentage of A21 (trapped water) but decreased the percentage of A22 (free water) in muscle tissue. Additionally, the microstructural results indicated that soaking in xylitol and mannitol significantly slowed the damage caused to muscle tissues by large ice crystals. Here, the incorporated xylitol and mannitol molecules presumably replaced water molecules by forming large hydrogen bonds with the muscle proteins, thereby stabilizing their structures in the absence of water while in the frozen state and suppressing the destruction of tissue structure. To note, the lactitol, isomaltitol, and maltitol used in this study did not seem to significantly contribute to cryoprotective effects on shrimp muscle during the experimental period.

In the present study, a tetrahedron probe with encoded internal reference nanoparticles (NPs) was self-assembled by a complementary nucleic acid aptamer for simultaneous ratiometric detection of telomerase (TE) and epithelial cell-adhesion molecule (EpCAM) in living cells. In the presence of a target, the dissociation of gold (Au) NPs, which was modified with corresponding tags, resulted in decreased surface-enhanced Raman scattering (SERS) signals. In addition, the ratios of Raman intensity at 1346 cm-1/1096 cm-1 (TE) and 1614 cm-1/1096 cm-1 (EpCAM) compared with the internal reference were demonstrated to quantify the level of TE and EpCAM, respectively, and can eliminate certain background noise. A good linear relationship was observed between them, and the linear range of TE and EpCAM in HeLa cells was 0.7 × 10-12 to 37.5 × 10-12 IU and 1.24 to 75.48 pg/mL with a limit of detection (LOD) of 7.6 × 10-16 IU and 0.53 pg/mL, respectively, which were consistent with the results of Raman confocal imaging. Meanwhile, the versatility and specificity of the developed probes were confirmed in cell lines. These results provide a reliable and ultrasensitive strategy for the in situ detection of biomarkers and a new method for SERS-based tetrahedrons in the early diagnosis of cancer.

BackgroundImported malaria has been an important challenge for China. Fatality rates from malaria increased in China, particularly in Henan Province, primarily due to malpractice and misdiagnoses in healthcare institutions, and the level of imported malaria. This study aims to investigate the relationship between the state of diagnosis and subsequent complications among imported malaria cases at healthcare institutions, based on malaria surveillance data in Henan Province from 2012 to 2017.MethodsA retrospective descriptive analysis was performed using data from the Centre for Disease Control and Prevention, Zhengzhou City, the capital of Henan Province. A decision tree method was exploited to provide valuable insight into the correlation between imported malaria cases and healthcare institutions.ResultsFrom 2012 to 2017, there were 371 imported malaria cases, mostly in males aged between 20 and 50 years, including 319Plasmodium falciparumcases. First visits of 32.3%, 19.9% and 15.9% malaria cases for treatment were to provincial, municipal and county healthcare institutions, respectively. The time interval between onset and initial diagnosis of 284 cases (76.5%) and the time interval between initial diagnosis and final diagnosis of 197 cases (53.1%) was no more than 72 h. An apparent trend was found that there were notably fewer patients misdiagnosed at first visit to healthcare institutions of a higher administrative level; 12.5% of cases were misdiagnosed in provincial healthcare institutions compared to 98.2% in private clinics, leading to fewer complications at healthcare institutions of higher administrative level due to correct initial diagnosis. In the tree model, the rank of healthcare facilities for initial diagnosis, and number of days between onset and initial diagnosis, made a major contribution to the classification of initial diagnosis, which subsequently became the most significant factor influencing complications developed in the second tree model. The classification accuracy were 82.2 and 74.1%, respectively for the tree models of initial diagnosis and complications developed.ConclusionInadequate seeking medical care by imported malaria patients, and insufficient capacity to diagnose malaria by healthcare institutions of lower administrative level were identified as major factors influencing complications of imported malaria cases in Henan Province. The lack of connection between uncommon imported malaria cases and superior medical resources was found to be the crucial challenge. A web-based system combined with WeChat to target imported malaria cases was proposed to cope with the challenge.

While no significant differences in initial ovule number were found among oilseed rape genotypes, there was a large variation in effective ovule number (EON), which determines the final seeds per silique (SPS), a critical component of yield. In this study, we selected 18 oilseed rape genotypes with contrasting nitrogen utilization efficiency (NUtE) to unravel the main factors responsible for different EON and determine the critical period of EON formation under both a field and a pot experiments from 2016-2018. The high NUtE genotypes displayed significantly higher NUtE by 14.3%, along with greater yield per plant (29.4%) and SPS (21.1%) than the low NUtE genotypes. The greater productivity of the high NUtE genotypes was associated with 44.1% higher pollen grain number, 23.5% greater pollen viability, and 39.3% lower ovule abortion rate, compared to the low NUtE genotypes. In addition, the heart stage was the critical ovule development period for delineating the variability of EON among contrasting NUtE oilseed rape genotypes, when the high NUtE genotypes displayed higher silique net photosynthetic rate, surface area, biomass, and RNA expression levels. Taken together, this study indicated the pollen grain number, pollen viability and ovule abortion rate contributed to the final variation in EON and the heart stage was the critical period of determining the EON differences among contrasting NUtE genotypes. Increasing pollen grain number and pollen viability, and decreasing ovule abortion rate before heart stage should be the prerequisite for breeders to improve yield and NUtE of oilseed rape genotypes.

The external and internal factors of the cell are critical to glioma initiation. Several factors and molecules have been reported to be implicated in the initiation and progression of brain cancer. However, the exact sequence of events responsible for glioma initiation is still unknown. Existing reports indicate that glioma stem cells are the cell of glioma origin. During cell division, chromosome breakage, DNA alteration increases the chance of cell genome modifications and oncogene overexpression. Although there is a high risk of gene alteration and oncogene overexpression, not everyone develops cancer. During embryogenesis, the same oncogenes that promote cancers have also been reported to be highly expressed, but this high expression which does not lead to carcinogenesis raises questions about the role of oncogenes in carcinogenesis. The resistance of cancer cells to drugs, apoptosis, and immune cells does not rely solely on oncogene overexpression but also on the defect in cell organelle machinery (mitochondria, endoplasmic reticulum, and cytoskeleton). This review discusses factors contributing to cancer; we report the dysfunction of the cell organelles and their contribution to carcinogenesis, while oncogene overexpression promotes tumorigenesis, maintenance, and progression through cell adhesion. All these factors together represent a fundamental requirement for cancer and its development.

Colorectal cancer (CRC) is a common cancer worldwide with complex etiology. Fusobacterium nucleatum (F. nucleatum), an oral symbiotic bacterium, has been linked with CRC in the past decade. A series of gut microbiota studies show that CRC patients carry a high abundance of F. nucleatum in the tumor tissue and fecal, and etiological studies have clarified the role of F. nucleatum as a pro-carcinogenic bacterium in various stages of CRC. In this review, we summarize the biological characteristics of F. nucleatum and the epidemiological associations between F. nucleatum and CRC, and then highlight the mechanisms by which F. nucleatum participates in CRC progression, metastasis, and chemoresistance by affecting cancer cells or regulating the tumor microenvironment (TME). We also discuss the research gap in this field and give our perspective for future studies. These findings will pave the way for manipulating gut F. nucleatum to deal with CRC in the future.

Phragmites australis is a cosmopolitan plant species with high intraspecific diversity and phenotypic plasticity. Due to its variability and large ecological niche breadth, subgroups of P. australis have become invasive in North America, and this invasion has been recognized late. While this cryptic invasion on the American continent has received much attention, little is known about the potential invasiveness of other subgroups, especially within Asian/Australian P. australis. We therefore compared the performance of three subgroups within the Asian/Australian group: a freshwater (CN) and an estuarine (YRD) subgroup collected in China and a genetically closely related subgroup collected from Australia (FEAU), grown in two common gardens in China. Our results showed that the FEAU subgroup had no strong invasive potential, as its total biomass, height, shoot number, specific leaf area, and stomatal conductance were lower than that of the two native subgroups. All three subgroups responded similarly with most traits to the different climates of the gardens, albeit with different response strength, expressed as phenotypic plasticity indices. The potential cryptic invasion risk of the FEAU subgroup in China seems to be low, since its functional traits showed low competitiveness and most traits with the lowest plasticity occurred in FEAU. However, caution is still advised, because other invasive mechanisms, such as enemy release or the performance under extreme environmental conditions were not tested in our study.

5-Methylcytosine (m5C) is a widespread post-transcriptional RNA modification and is reported to be involved in manifold cellular responses and biological processes through regulating RNA metabolism. However, its regulatory role in antiviral innate immunity has not yet been elucidated. Here, we report that NSUN2, a typical m5C methyltransferase, can negatively regulate type I interferon responses during viral infection. NSUN2 specifically mediates m5C methylation ofIRF3mRNA and accelerates its degradation, resulting in low levels of IRF3 and downstream IFN-β production. Knockout or knockdown of NSUN2 could enhance type I interferon responses and downstream ISG expression after viral infectionin vitro. Andin vivo, the antiviral innate responses is more dramatically enhanced inNsun2+/−mice than inNsun2+/+mice. Four highly m5C methylated cytosines inIRF3mRNA were identified, and their mutation could enhance the cellularIRF3mRNA levels. Moreover, infection with Sendai virus (SeV), vesicular stomatitis virus (VSV), herpes simplex virus 1 (HSV-1), Zika virus (ZIKV), or especially SARS-CoV-2 resulted in a reduction in endogenous levels of NSUN2. Together, our findings reveal that NSUN2 serves as a negative regulator of interferon response by accelerating the fast turnover ofIRF3mRNA, while endogenous NSUN2 levels decrease after viral infection to boost antiviral responses for the effective elimination of viruses. Our results suggest a paradigm of innate antiviral immune responses ingeniously involving NSUN2-mediated m5C modification.

The complete chloroplast genome sequence of Catalpa fargesii f. duclouxii C.A. May was firstly assembled and analyzed in this study. The whole genome of this species was 158164 bp in length, with a...

Musk Larkspur (Delphinium brunonianum) is a perennial herb of the family Ranunculace with medicinal values. In this study, the chloroplast (cp) genome of this herb was determined to be 153,926 bp long with an A + T-biased base composition, and comprises a pair of inverted repeat (IR) regions (26,559 bp), separated by a large single-copy (LSC) region (84,512 bp) and a small single-copy (SSC) region (16,296 bp). A total of 112 gene species were annotated with 19 of them being completely or partially duplicated. Eighteen gene species harbor one or two introns. Phylogenetic analysis challenged the monophyly of the subfamily Ranunculoideae.

High Fischer ratio oligo-peptides hydrolysate (DAHFP) from minced meat of Skipjack (Katsuwonus pelamis) were prepared by hydrolysis of pepsin and flavourzyme, then adsorpte on activated car...

AIM: The integration of palaeontological and phylogenetic data can improve our understanding in the spatio‐temporal evolutionary processes of living organisms. However, how best to use fossil data in divergence time estimation and ancestral range reconstruction remains challenging. Here, we integrated palaeontological and molecular data to investigate the historical biogeography of Hamamelidaceae, a pantropical angiosperm family with abundant fossils outside its present distribution. LOCATION: Global tropical/subtropical areas. METHODS: Using seven DNA regions (>7,500 bp) from plastid and nuclear genomes, we reconstructed a robust phylogenetic framework for Hamamelidaceae with the first complete genus‐level sampling. We used the tip‐dating method with the 22 fossils to estimate divergence times for the family, and inferred the ancestral range of lineages under the dispersal–extinction–cladogenesis model by incorporating the fossils. RESULTS: Our biogeographic analysis indicates that extant Hamamelidaceae most likely originated in tropical Asia during the mid‐Cretaceous, and in the family 20 dispersals occurred during three major time intervals: the Upper Cretaceous (c. 93–69 Ma), Palaeocene–Eocene (c. 63–39 Ma) and late Oligocene (c. 27–23 Ma). MAIN CONCLUSIONS: Overland migrations through available land bridges and island chains may have been mainly responsible for hamamelidaceous range expansions during these three episodes. This study contributes to our knowledge on the assembly and evolution of angiosperm‐dominated tropical and subtropical forests.

Background Lenvatinib is a newly approved molecular targeted drug for the treatment of advanced hepatocellular carcinoma (HCC). However, the high cost associated with this treatment poses a huge financial burden on patients and the entire public health system. Therefore, there is an urgent need to develop novel strategies that enhance the antitumor effect of lenvatinib. Methods The antitumor effects of chelidonine or/and lenvatinib on HCC cell lines MHCC97-H and LM-3 were examined using the 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2- H-tetrazolium bromide (MTT) assay. For the in-vivo investigation, the effect on subcutaneous or intrahepatic tumor growth in nude mice was also determined. The mRNA levels of epithelial mesenchymal transition (EMT)-related factors were examined through quantitative polymerase chain reaction or Western blot. Results In the present study, we found that treatment with chelidonine enhanced the apoptotic effect of lenvatinib on HCC cells and the in-vivo growth of HCC tumors in nude mice. Mechanistically, treatment with chelidonine increased the expression of epithelial indicator E-cadherin, whereas it decreased the expression of mesenchymal indicators N-cadherin and Vimentin. These findings suggest that chelidonine restricted the EMT in HCC cells. Conclusion Chelidonine inhibits the process of EMT and enhances the antitumor effect of lenvatinib on HCC cells.

Summary Traditional approaches for sequencing insertion ends of bacterial artificial chromosome (BAC) libraries are laborious and expensive, which are currently some of the bottlenecks limiting a better understanding of the genomic features of auto‐ or allopolyploid species. Here, we developed a highly efficient and low‐cost BAC end analysis protocol, named BAC‐anchor, to identify paired‐end reads containing large internal gaps. Our approach mainly focused on the identification of high‐throughput sequencing reads carrying restriction enzyme cutting sites and searching for large internal gaps based on the mapping locations of both ends of the reads. We sequenced and analysed eight libraries containing over 3 200 000 BAC end clones derived from the BAC library of the tetraploid potato cultivar C88 digested with two restriction enzymes, Cla I and Mlu I. About 25% of the BAC end reads carrying cutting sites generated a 60–100 kb internal gap in the potato DM reference genome, which was consistent with the mapping results of Sanger sequencing of the BAC end clones and indicated large differences between autotetraploid and haploid genotypes in potato. A total of 5341 Cla I‐ and 165 Mlu I‐derived unique reads were distributed on different chromosomes of the DM reference genome and could be used to establish a physical map of target regions and assemble the C88 genome. The reads that matched different chromosomes are especially significant for the further assembly of complex polyploid genomes. Our study provides an example of analysing high‐coverage BAC end libraries with low sequencing cost and is a resource for further genome sequencing studies.

Background Cutaneous melanoma is the most aggressive form of skin cancer. It accounts for approximately 5% of all cutaneous malignancies and is currently responsible for the majority of skin cancer-related deaths. However, the exact mechanisms responsible for the occurrence of melanoma, in particular the invasive growth in normal skin or muscle tissue, remain unknown. Materials and methods miR-3662, a microRNA is a potential tumor suppressor targeting zinc finger E-box binding homeobox 1 (ZEB1), which functions as a key regulator of the epithelial-mesenchymal transition (EMT) process. This microRNA was identified using an online database (miRDB) and expression was confirmed by Western blot analysis. Quantitative polymerase chain reaction (qPCR) was used to examine whether miR-3662 inhibits the EMT process in the aggressive melanoma cell line, A375, through the modification of the expression of invasion-related genes in A375 cells. The effects of miR-3662 on the in vivo growth of A375 cells were examined in a nude mouse model. Results Using virtual screening of the miRDB database, miR-3662 was shown to target the 3' untranslated region (UTR) of the ZEB1 gene. Expression of miR-3662 via a lentivirus vector significantly decreased protein levels of ZEB1 and inhibited the growth of A375 cells in vitro and in vivo. The reduction in ZEB1 expression induced by miR-3662 resulted in EMT inhibition in A375 cells and decreased the relative expression of metastasis genes. Conclusion Down-regulation of ZEB1's expression via miR-3662 lentivirus vectors significantly decreased the in vitro and in vivo growth of the highly aggressive melanoma cell line A375.

Hemiboeaalbiflora X.G.Xiang, Z.Y.Guo & Z.W.Wu, sp. nov., a new species of Gesneriaceae from Guizhou, China, is described and illustrated. This species was previously listed informally as a variety of H.gamosepala, but it differs significantly from H.gamosepala by its 5-parted calyx from the base, longer peduncle, white corolla and longer pistil. Based on recent extensive observations, this new species is similar to H.cavalerieivar.paucinervis and H.subcapitata but differs from them by its longer petiole, larger involucre, white corolla and longer staminal filaments. The conservation status of this species is considered to be “Vulnerable” (VU) according to the IUCN Red List Categories and Criteria.

Fruit length and diameter are very important appearance quality which affects preference of consumers and quality of fruit. To improve the genetic mechanisms of cucumber fruit shape and size, an F2:5 recombinant inbred line (RIL)-population was derived from a cross between Q16 inbred lines which had slender fruit and Q24 inbred lines which had round fruit to screen fruit shape QTLs locus. 85 RILs, 15 F1 and 15 of each parental line were grown in the spring of 2016 and 2017 respectively. The length and diameter of ovary, immature fruit and mature fruit were measured at blooming day, 12 days after pollination and 35 days after pollination respectively. The RIL population was screened basing on the data related fruit using 89 SSR markers and 1 InDel marker. Twenty-one QTLs loci were totally identified for fruit shape traits from RIL population. Comparing the flanking markers of 21 QTL loci allowed to 5 consensus QTLs FS1.1, FS1.2, FS2.1, FS3.1 and FS6.1 loci, in which FS3.1 played a role in fruit elongation, FS6.1 was important for the increase of fruit diameter, and the else 3 QTLs acted in both fruit elongation and diameter. The results from this study may increase the understanding of the genetic basis control of fruit shape and size and provide support for marker-assisted selection in cucumber with ideal fruit shape.

Embryology can provide valuable information for determining taxonomic and phylogenetic relationships at the genus level. However, embryological information is fragmentary for the genus of Clematis. Herein, we studied the anther and ovule development of Clematis terniflora var. mandshurica using paraffin sections to examine microsporogenesis, microgametogenesis, megasporogenesis and macrogametogenesis in this plant. Tetrasporangiate anther, successive microsporocyte cytokinesis, and glandular tapetum, or occasionally amoeboid tapetum were observed in C. terniflora var. mandshurica. This plant’s microspore tetrads were mostly tetrahedral, and occasionally symmetrical, after meiosis. The anther wall was comprised of an epidermis and endothecium, both with fibrous thickenings in the mature anthers. Pollen grains were symmetrical, oblate, tricolpate, and two-celled. As for the ovary, it had one chamber with a few degraded ovules and a single normal one, which was anatropous, unitegmic, and tenuinucellate, and formed a Polygonum-type embryo-sac. Only one archesporial cell was present, and developed directly into a megaspore mother cell, which later formed a linear tetrad of megaspores. Larger dikaryocytic antipodal cells could also be observed in the mature embryo sac. This species might belong to a relatively primitive and transitional clade within the genus Clematis. To our knowledge, this study documented the embryological characters of C. terniflora var. mandshurica in detail for the first time and provided more information on the embryology of the genus Clematis.

An efficient double-chitinase hydrolysis of crab shell particles (CSP) pretreated by ionic liquids was investigated in this work. First, two family 18 chitinase genes (chip1 and chip2) from Paeniba...

Nitrogen (N) is an essential macronutrient for plant development and growth, and the deposition of N has increased in recent decades. Legumes that fix N can also provide N for nearby species. However, N in soil inhibits N fixation. We tested the effects of N fertilisation on one N-fixing (Robinia pseudoacacia) and two non-N-fixing (Sophora japonica and Senna surattensis) woody legume species, which were subjected to five different N levels (0, 1.5, 2.9, 5.9 and 11.4 mg N per plant day−1) under greenhouse conditions. The growth of the two non-N-fixing species was promoted by N supply, while that of R. pseudoacacia was unaffected. Among the three species, R. pseudoacacia had the largest specific leaf area and chlorophyll concentration, S. japonica had the largest root-to-shoot ratio and main root-to-lateral root ratio, and S. surattensis had the largest leaf N and phosphorus concentrations. The N-fixing species was mostly unaffected by N supply. The growth, leaf chlorophyll concentration, and leaf number in the non-N-fixing species were promoted by N supply. The N-fixing species showed better growth in low-N environments, while under increased N deposition, its growth was similar to that of the non-N-fixing species.