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SRS (SHI-related sequence) transcription factors play a crucial role in plant growth, development, and abiotic stress response. Although Brassica napus (B. napus) is one of the most important oil crops in the world, the role of SRS genes in B. napus (BnSRS) has not been well investigated. Therefore, we employed a bioinformatics approach to identify BnSRS genes from genomic data and investigated their characteristics, functions, and expression patterns, to gain a better understanding of how this gene family is involved in plant development and growth. The results revealed that there were 34 BnSRS gene family members in the genomic sequence of B. napus, unevenly distributed throughout the sequence. Based on the phylogenetic analysis, these BnSRS genes could be divided into four subgroups, with each group sharing comparable conserved motifs and gene structure. Analysis of the upstream promoter region showed that BnSRS genes may regulate hormone responses, biotic and abiotic stress response, growth, and development in B. napus. The protein-protein interaction analysis revealed the involvement of BnSRS genes in various biological processes and metabolic pathways. Our analysis of BnSRS gene expression showed that 23 BnSRS genes in the callus tissue exhibited a dominant expression pattern, suggesting their critical involvement in cell dedifferentiation, cell division, and tissue development. In addition, association analysis between genotype and agronomic traits revealed that BnSRS genes may be linked to some important agronomic traits in B. napus, suggesting that BnSRS genes were widely involved in the regulation of important agronomic traits (including C16.0, C18.0, C18.1, C18.2 C18.3, C20.1, C22.1, GLU, protein, TSW, and FFT). In this study, we predicted the evolutionary relationships and potential functions of BnSRS gene family members, providing a basis for the development of BnSRS gene functions which could facilitate targeted functional studies and genetic improvement for elite breeding in B. napus.

Oilseed rape (Brassica napus L.), as an important source of renewable energy and an industrial crop, has received extensive attention. Breeding cultivars with appropriate flowering time (FT) is of high significance for crop yields, but it is currently impeded by incomplete understanding of genetic control for flowering time in B. napus. In this study, ebm, an ethyl methanesulfonate mutagenesis (EMS) mutant with early blooming, was derived from B. napus cv. Zhongshuang9 (ZS9). Genetic analysis showed that the early blooming of ebm was controlled by a dominant nuclear gene. The Ebm locus was delimitated in a 905-kb region on chromosome A02 by bulked segregant analysis sequencing and fine mapping. Based on the expression and variation analysis of the genes in the candidate region, and gene conservation analysis in pan-genome, a potential gene BnaA02.CRF10 was pinpointed, which encodes a cytokinin response factor involved in the cytokinin signaling pathway. Regional association and haplotype analyses of the initial and blooming FT in a worldwide bio-panel of 263 re-sequenced B. napus accessions further confirmed the candidate gene and mined favorable allelic variations. Additionally, differentially expressed genes (DEGs) related to ethylene and cytokinin genetic factors were significantly induced in the ebm mutant by comparative transcriptome analysis. Moreover, phylogenetic and synteny network analyses provided strong evidence for the phylogenetic conservation of the CRF subfamily during the evolutionary history from Chlorophyta to dicotyledonous plants. Taken together, a novel conserved gene with favorable alleles conferring early flowering trait in B. napus was identified, which lays a foundation for the genetic improvement and cultivar breeding of this important crop with shortened growth periods in the future.

Oilseed rape (Brassica napus) with yellow flowers is an attractive ornamental landscape plant during the flowering period, and the development of different petal colors has become a breeding objective. Although yellowish flower color is a common variant observed in field-grown oilseed rape, the genetics behind this variation remains unclear. We obtained a yellowish-white flower (ywf) mutant from Zhongshuang 9 (ZS9) by ethyl methanesulfonate mutagenesis (EMS) treatment. Compared with ZS9, ywf exhibited a lower carotenoid content with a reduced and defective chromoplast ultrastructure in the petals. Genetic analysis revealed that the yellowish-white trait was controlled by a single recessive gene. Using bulked-segregant analysis sequencing (BSA-seq) and kompetitive allele-specific PCR (KASP), we performed map-based cloning of the ywf locus on chromosome A08 and found that ywf harbored a C-to-T substitution in the coding region, resulting in a premature translation termination. YWF, encoding phytoene desaturase 3 (PDS3), was highly expressed in oilseed rape petals and involved in carotenoid biosynthesis. Pathway enrichment analysis of the transcriptome profiles from ZS9 and ywf indicated the carotenoid biosynthesis pathway to be highly enriched. Further analyses of differentially expressed genes and carotenoid components revealed that the truncated BnaA08.PDS3 resulted in decreased carotenoid biosynthesis in the mutant. These results contribute to an understanding of the carotenoid biosynthesis pathway and manipulation of flower-color variation in B. napus.

Background Oilseed rape (Brassica napus L.) is known as one of the most important oilseed crops cultivated around the globe. However, in the humid season, it faced a severe challenge of Sclerotinia stem rot (SSR), a severely destructive disease caused by the fungus Sclerotinia sclerotiorum, which is negatively impact in terms of huge reduction in yield loss and annually. Like other field crops, in Brassica napus (B. napus) SSR resistance is quantitatively controlled, and there are few studies reported the effective major genes conferring SSR disease resistance so far. Thus, there is utmost need to design the studies at whole genome level to discover the promising major genes, which can further enhance the protective shield against SSR in B. napus. Results Here, we used a natural population of B. napus consisting of 222 accessions to perform a genome-wide association study (GWAS) to find the candidate genes conferencing the SSR resistance trait. Our results showed that a total of 2779265 SNP markers were identified, which harboring 51 candidate genes. Moreover, we found that candidate gene BnaA08g25340D (BnMLO2_2), a member of seven homolog genes of Arabidopsis Mildew Locus O 2 (MLO2), was closely associated with the SSR resistance. The natural SNP variations was predominantly identified in the promoter region of BnMLO2_2 and 3 haplotypes were found to be closely related to the differential expressions of BnMLO2_2 in the leaves of the B. napus population, which may responsible for the variations of SSR resistance observed in this population. Besides, we also noted that among seven members of MLO2, only BnMLO2_2 showed constitutive expression in the leaf tissues. In Arabidopsis, loss of function mutation in MLO2 displayed enhanced susceptibility to SSR, whereas, Arabidopsis plants overexpression of MLO2 genes showed significantly enhanced resistance to SSR disease. The mechanism underlying MLO2 in the regulation of SSR resistance was associated with the cell death. Lastly, syntenic analysis revealed 57, 23, and 28 homolog genes of 15 Arabidopsis MLOs in the genomes of B. napus, Brassica rapa (B. rapa), and Brassica oleracea (B. oleracea), respectively indicated an asymmetrical evolution between the A and C subgenomes of B. napus. Conclusion Present investigation revealed the roles of MLO2 gene in the regulation of SSR disease resistance. Therefore, natural variations in the promoter region of BnMLO2 gene can be utilized for improving the resistance to SSR disease in B. napus. MLO locus play antagonistic role against SSR and PM diseases.

The trans-Golgi network (TGN) acts as a central platform for sorting and secreting various cargoes to the cell surface, thus being essential for the full execution of plant immunity. However, the fine-tuned regulation of TGN components in plant defense and stress response has been not fully elucidated. Our study revealed that despite largely compromising penetration resistance, the loss-of-function mutation of the TGN component protein ECHIDNA (ECH) induced enhanced postinvasion resistance to powdery mildew in Arabidopsis thaliana. Genetic and transcriptome analyses and hormone profiling demonstrated that ECH loss resulted in salicylic acid (SA) hyperaccumulation via the ISOCHORISMATE SYNTHASE 1 biosynthesis pathway, thereby constitutively activating SA-dependent innate immunity that was largely responsible for the enhanced postinvasion resistance. Furthermore, the ech mutant displayed accelerated SA-independent spontaneous cell death and constitutive POWDERY MILDEW RESISTANCE 4-mediated callose depositions. In addition, ECH loss led to a chronically prolonged endoplasmic reticulum stress in the ech mutant. These results provide insights into understanding the role of TGN components in the regulation of plant immunity and stress responses.

As a soilborne protist pathogen, Plasmodiophora brassicae causes the devastating clubroot disease on Brassicaceae crops worldwide. Due to its intracellular obligate biotrophic nature, the life cycle of P. brassicae is still not fully understood. Here, we used fluorescent probe-based confocal microscopy and transmission electron microscopy (TEM) to investigate the infection process of P. brassicae on the susceptible host Arabidopsis under controlled conditions. We found that P. brassicae can initiate the primary infection in both root hairs and epidermal cells, producing the uninucleate primary plasmodium at 1 day postinoculation (dpi). After that, the developed multinucleate primary plasmodium underwent condensing and cytoplasm cleavage into uninucleate zoosporangia from 1 to 4 dpi. This was subsequently followed by the formation of multinucleate zoosporangia and the production of secondary zoospores within zoosporangium. Importantly, the secondary zoospores performed a conjugation in the root epidermal cells after their release. TEM revealed extensive uninucleate secondary plasmodium in cortical cells at 8 dpi, indicating the establishment of the secondary infection. The P. brassicae subsequently developed into binucleate, quadrinucleate, and multinucleate secondary plasmodia from 10 to 15 dpi, during which the clubroot symptoms appeared. The uninucleate resting spores were first observed in the cortical cells at 24 dpi, marking the completion of a life cycle. We also provided evidence that the secondary infection of P. brassicae may represent the diploid sexual life stage. From these findings, we propose a refined life cycle of P. brassicae which will contribute to understanding of the complicated infection biology of P. brassicae.

The actin cytoskeleton regulates an array of diverse cellular activities that support the establishment of plant–microbe interactions and plays a critical role in the execution of plant immunity. However, molecular and cellular mechanisms regulating the assembly and rearrangement of actin filaments (AFs) at plant–pathogen interaction sites remain largely elusive. Here, using live-cell imaging, we show that one of the earliest cellular responses in Arabidopsis thaliana upon powdery mildew attack is the formation of patch-like AF structures beneath fungal invasion sites. The AFs constituting actin patches undergo rapid turnover, which is regulated by the actin-related protein (ARP)2/3 complex and its activator, the WAVE/SCAR regulatory complex (W/SRC). The focal accumulation of phosphatidylinositol-4,5-bisphosphate at fungal penetration sites appears to be a crucial upstream modulator of the W/SRC–ARP2/3 pathway-mediated actin patch formation. Knockout of W/SRC–ARP2/3 pathway subunits partially compromised penetration resistance with impaired endocytic recycling of the defense-associated t-SNARE protein PEN1 and its deposition into apoplastic papillae. Simultaneously knocking out ARP3 and knocking down the Class I formin (AtFH1) abolished actin patch formation, severely impaired the deposition of cell wall appositions, and promoted powdery mildew entry into host cells. Our results demonstrate that the ARP2/3 complex and formins, two actin-nucleating systems, act cooperatively and contribute to Arabidopsis penetration resistance to fungal invasion.

Background This cohort study was designed to investigate the prevalence of and potential risk factors of HEV infection in a large multi-ethnic youth cohort in China. Methods Blood samples were collected from participants (n = 6269) and serum was isolated. All serum samples were tested for anti-HEV IgG, anti-HEV IgM antibodies using commercial enzyme immunoassay kits (Wantai Biological Pharmacy Enterprise, Beijing, China). Results The overall rate of anti-HEV IgG and anti-HEV IgM prevalence was 4.78% and 0.14%, 0.03% were positive for both anti-HEV IgG and anti-HEV IgM antibodies. Anti-HEV IgG positivity is significantly higher in females (5.27%) compared to males (4.14%) (P = 0.028). Anti-HEV IgG prevalence is significantly (P = 0.0001) higher in Dong (17.57%), Miao (12.23%), Yi (11.04%), Gelao (9.76%), and Bai (10.00%) compared to other ethnic groups. It is significantly higher in Guizhou (11.4%), Sichuan (10.1%), Yunnan (9.3%), and Guangxi (6.9%) than that other province. We found that ethnicity and provincial background are significantly associated with HEV infection in this cohort. Conclusion This study provides comprehensive information on HEV prevalence in multi-ethnic populations in China. However, our study only focused on a youth population from different provinces of China. Future studies are recommended to investigate HEV prevalence in other age groups of the ethnic populations.

Additional file 1. Participants provincial background.

The completion of B. oleracea genome sequencing has greatly enhanced Brassica research, especially for revealing genome structure and evolution of polyploidy. Based on intraspecies syntenic analysis and comparison with B. rapa, the asymmetrical evolution of polyploid Brassica genomes were elucidated at the DNA, RNA, and epigenetic levels. Main features include asymmetrical gene loss in chromosomal blocks derived from whole-genome triplication (WGT), asymmetrical sequence, and expression divergence among WGT-derived duplicated genes, asymmetrical 24-nt sRNA regulated subgenome expression dominance, asymmetrical TE (transposable elements) accumulation, gene expansion between two Brassica genomes, and asymmetrical contribution to the novelty of agronomic traits between the different subgenomes. It was found that the multilayered asymmetrical evolution is also observed in ancient polyploids such as Arabidopsis, maize, grape, soybean, grasses as well as recent allopolyploids such as rapeseed, cotton, and wheat. These evidences support the role of asymmetrical evolution in the evolutionary success of polyploidy as well as their phenotypic novelty and adaptation. Further understanding of the mechanism of asymmetrical evolution of polyploid genomes lays a foundation for genetic breeding and enhancement of elite cultivars.

BackgroundThis paper aimed to explore the average natural menopause age, the span of menopausal age, and the related factors that will probably affect the natural menopause age of Chinese women. MethodsA large-scale random sampling of menopausal women aged 32-75 years in 18 provinces or municipalities throughout China was conducted using paper questionnaire surveys to investigate the menopause age, menarche age, fertility status, illness history, and living habits. The SPSS 24.0 software package was used to carry out t test, chi-square test, logistic regression, and other statistical analyses. ResultsA total of 5852 questionnaires were collected. After excluding questionnaires having no menopause, questionnaires with incomplete or invalid information, and questionnaires that may directly affect the age of menopause, such as hysterectomy, oophorectomy, and other factors that may affect natural menopause, a total of 4426 effective questionnaires have been collected, and the average age of natural menopause was 49.49 ± 3.67 years old. ConclusionFactors that may affect the age of natural menopause are weight, menarche age, marital status, drinking tea or not (often and daily), eating supplements, oral contraceptive contraception, and safe contraception.

As a soilborne protist pathogen

Estrogen evidently exerts a protective role against gastric cancer. Accordingly, we evaluated the relationship between the expression of the estrogen receptor ER-α36 and the clinicopathologic features in gastric cancer. ER-α36 expression levels differed among the tumor center, invasion front, and vascular metastases. The effects of E2β (17β-Estradiol, E2β) on invasion ability in SGC7901, High36 (with ER-α36 upregulation), and Low36 (with ER-α36 downregulation) cells were evaluated using Transwell assays. Furthermore, the c-Src signaling pathway was inhibited using PP2 and the effects on E2β-induced increases in E-cadherin, MMP2, and MMP9 were evaluated using western blotting. ER-α36, c-Src, MMP2, and E-cadherin levels were also evaluated in tumor xenografts. We found that 0.1 nM E2β promoted gastric cancer cell invasion by reducing E-cadherin expression and increasing MMP2 and MMP9 levels. The upregulation of ER-α36 promoted gastric cancer cell invasion and the downregulation of ER-α36 reduced the invasive ability of cells. The levels of ER-α36, c-Src, and MMP2 were the highest in tumor xenografts using High36 cells, intermediate in tumor xenografts using SGC7901 cells, and lowest in tumor xenografts using Low36 cells. The opposite results were obtained for E-cadherin expression. ER-α36 enhanced gastric cancer cell invasion by the activation of membrane-initiated c-Src signaling pathways. In particular, treatment with E2β and ER-α36 influenced gastric cancer cell invasion. Furthermore, c-Src was involved in the ER-α36-mediated estrogen signaling pathway and cell invasion.

Different phosphoinositides enriched at the membranes of specific subcellular compartments within plant cells contribute to organelle identity, ensuring appropriate cellular trafficking and function. During the infection of plant cells, biotrophic pathogens such as powdery mildews enter plant cells and differentiate into haustoria. Each haustorium is enveloped by an extrahaustorial membrane (EHM) derived from the host plasma membrane. Little is known about the EHM biogenesis and identity. Here, we demonstrate that among the two plasma membrane phosphoinositides in Arabidopsis (Arabidopsis thaliana), PI(4,5)P2 is dynamically up-regulated at powdery mildew infection sites and recruited to the EHM, whereas PI4P is absent in the EHM. Lateral transport of PI(4,5)P2 into the EHM occurs through a brefeldin A-insensitive but actin-dependent trafficking pathway. Furthermore, the lower levels of PI(4,5)P2 in pip5k1 pip5k2 mutants inhibit fungal pathogen development and cause disease resistance, independent of cell death-associated defenses and involving impaired host susceptibility. Our results reveal that plant biotrophic and hemibiotrophic pathogens modulate the subcellular distribution of host phosphoinositides and recruit PI(4,5)P2 as a susceptibility factor for plant disease.

Bradykinin (BK) promotes insulin sensitivity and glucose uptake in adipocytes and other cell types. We demonstrated that in rat adipocytes BK enhances insulin-stimulated glucose transport via endothelial nitric oxide synthase, nitric oxide (NO) generation, and decreased activity of the mitogen-activated protein kinase (MAPK) JNK (c-Jun NH2-terminal kinase). In endothelial cells, NO increases soluble guanylate cyclase (sGC) activity, which, in turn, activates protein kinase G (PKG) by increasing cGMP levels. In this study, we investigated whether BK acts via the sGC-cGMP-PKG pathway to inhibit the negative effects of JNK on insulin signaling and glucose uptake in rat adipocytes. BK augmented cGMP concentrations. The BK-induced enhancement of insulin-stimulated glucose uptake was mimicked by the sGC activator YC-1 and a cell-permeable cGMP analog, CPT-cGMP, and inhibited by the sGC inhibitor ODQ and the PKG inhibitor KT 5823. Transfection of dominant-negative PKG reduced the BK augmentation of insulin-induced Akt phosphorylation. The activation of JNK and ERK1/2 by insulin was attenuated by BK, which was mediated by the sGC-cGMP-PKG pathway. Whereas insulin-stimulated phosphorylation of upstream activators of JNK and ERK, i.e., MKK4 and MEK1/2, was unaffected, BK augmented insulin-mediated induction of MKP-5 mRNA and protein levels. Furthermore, zaprinast, a phosphodiesterase inhibitor, enhanced cGMP and MKP-5 and prolonged the action of BK. These data indicate that BK enhances insulin action by inhibition of negative feedback by JNK and ERK via upregulation of MKP-5, mediated by the sGC-cGMP-PKG signaling pathway.

A sub-class of ER-negative breast cancer that is negative for ER, PR and HER2 expression known as triple-negative breast cancer (TNBC) is highly malignant and lacks effective treatment. Recently, it has been reported that an isoform of estrogen receptor-alpha ER-α36 is expressed and plays a critical role in development of TNBC. ER-α36 forms a positive regulatory loop with epidermal growth factor receptor (EGFR), which promotes malignant growth of TNBC cells. Thus, ER-α36 has been proposed as an important target for development of novel drugs for TNBC. In this study, we evaluated the effects of icaritin, a prenylflavonoid derivant purified from Epimedium Genus, on growth of TNBC cells and examined the possible underlying mechanisms. Our study demonstrated that icartin decreased both ER-α36 and EGFR protein expression, and induced apoptosis in TNBC MDA-MB-231 and MDA-MB-453 cells. We also found that icaritin inhibited ER-α36-mediated MAPK/ERK pathway and cyclin D1 induction by estrogen. Our results thus indicated that icaritin has a potential to be developed into a novel therapeutic agent for human TNBC.

The aim of the present study was to investigate Fas expression in tumor samples from patients with gastric cancer, in order to determine the involvement of the Fas signaling pathway. The protein expression levels of Fas, caspase-8, caspase‑3 and poly (adenosine diphosphate‑ribose) polymerase 1 (PARP1) were examined in gastric cancer specimens and their associations with clinical pathological parameters were analyzed with immunohistochemical staining and western blot analysis. The mRNA expression was quantified with quantitative PCR and apoptosis was examined with a FACScan flow cytometer. The results demonstrated that the downregulation of Fas expression was correlated with less histological differentiation, gender (male), and increased lymph node and distant metastases (P

The potential infection biology of Plasmodiophora brassicae in resistant hosts and non-hosts is still not completely understood. Clubroot resistance assay on European clubroot differentials (ECD) set revealed that ECD10 (Brassica napus) and ECD4 (Brassica rapa) show a complete resistance to the tested P. brassicae isolate in contrast to highly susceptible hosts Westar (B. napus) and ECD5 (B. rapa). Previously, we used fluorescent probe-based confocal microscopy (FCM) to refine the life cycle of P. brassicae and indicate the important time points during its infection in Arabidopsis. Here, we used FCM to systematically investigate the infection of P. brassicae in two resistant host species ECD10 and ECD4 and two non-host crops wheat and barley at each indicated time points, compared with two susceptible hosts Westar and ECD5. We found that P. brassicae can initiate the primary infection phase and produce uninucleate primary plasmodia in both resistant hosts and non-hosts just like susceptible hosts at 2 days post-inoculation (dpi). Importantly, P. brassicae can develop into zoosporangia and secondary zoospores and release the secondary zoospores from the zoosporangia in resistant hosts at 7 dpi, comparable to susceptible hosts. However, during the secondary infection phase, no secondary plasmodium was detected in the cortical cells of both resistant hosts in contrast to massive secondary plasmodia present in the cortex tissue of two susceptible hosts leading to root swelling at 15 dpi. In both non-host crops, only uninucleate primary plasmodia were observed throughout roots at 7 and 15 dpi. Quantitative PCR based on DNA revealed that the biomass of P. brassicae has no significant increase from 2 dpi in non-host plants and from 7 dpi in resistant host plants, compared to the huge biomass increase in susceptible host plants from 2 to 25 dpi. Our study reveals that the primary infection phase in the root epidermis and the secondary infection phase in the cortex tissue are, respectively, blocked in non-hosts and resistant hosts, contributing to understanding of cellular and molecular mechanisms underlying clubroot non-host and host resistance.

The present study aimed to investigate the effects of 17β-estradiol and tamoxifen, an agonist and inhibitor of the estrogen receptor (ER), respectively, on the proliferation and apoptosis of gastric cancer cells, as well as the messenger (m)RNA expression levels of ER-α36. Nested reverse transcription-polymerase chain reaction (RT-PCR) confirmed that ER-α36 was expressed in the BGC823, MKN45 and SGC7901 human gastric cancer cell lines. Subsequently, the BGC823 cell line was stimulated with various concentrations of 17β-estradiol or tamoxifen for 24 or 48 h, and the proliferation, apoptosis and mRNA expression levels of ER-α36 were determined by water-soluble tetrazolium (WST)-1 assay, flow cytometry and RT-quantitative PCR, respectively. The activity of BGC823 cells was significantly increased following treatment with 10−12 mol/l 17β-estradiol for 24 h (P=0.013), as compared with the control, and reached a peak at 48 h (P=0.002). Notably, the activity of BGC823 cells was decreased with increasing concentrations of 17β-estradiol, although it remained higher compared with that of the control. In the tamoxifen-treated groups, the cell activity decreased as the drug concentration increased. The apoptosis rate was markedly reduced in the 17β-estradiol group after 24 h (10−12 mol/l, P=0.013; 10−11 mol/l, P=0.023; and 10−10 mol/l, P=0.017) and after 48 h (10−12 mol/l, P=0.002; 10−11 mol/l, P=0.011; and 10−10 mol/l, P=0.033), whereas the rate of apoptosis increased as the tamoxifen concentration increased (24 h: 5×10−6 mol/l, P=0.002; and 10−5 mol/l, P=0.001; and 48 h: 5×10−6 mol/l, P=0.014 and 10−5 mol/l, P=0.0021), as compared with the control group. The mRNA expression levels of ER-α36 were significantly increased after 24 h of treatment with 10−12 mol/l (P=0.024), 10−11 mol/l (P=0.0113) and 10−10 mol/l (P=0.0037) 17β-estradiol compared with the control group when the concentration of 17β-estradiol was low, and the same was observed after 48 h of treatment 10−12 mol/l (P=0.0164), 10−11 mol/l (P=0.0342) and 10−10 mol/l (P=0.0198) 17β-estradiol. The mRNA expression levels of ER-α36 were significantly decreased with increasing concentrations of tamoxifen after 24 h (5×10−6 mol/l, P=0.0233; and 10−5 mol/l, P=0.007) and after 48 h (5×10−6 mol/l, P=0.001; and 10−5 mol/l, P=0.0153). In addition, the ability of tamoxifen to inhibit the growth of gastric cancer cells was concentration-dependent. The results of the present study suggested that gastric cancer cells were sensitive to the effects of 17β-estradiol and tamoxifen, and that tamoxifen is able to induce gastric cancer cell apoptosis. The expression levels of ER-α36 were upregulated, and the growth of gastric cancer cells was increased, following treatment with 17β-estradiol, thus suggesting that gastric cancer tumors are stimulated by estrogen.

Puerto Rico suffered the worst hurricane which caused devastating damage in 2017. In order to deliver medical supplies in time and patrol the lines, NGOs try to improve the response capabilities of its by designing a portable disaster response system called DroneGo.DroneGo needs to find the optimal delivery path and the optimal equipment utilization rate for delivery according to the disaster relief conditions and schedule. In this paper, the integration of integer programming algorithm and Dijkstra algorithm is used to construct the shortest distribution path model for medical supplies transported by rescue drones in disaster areas. Using integer programming method, 7 kinds of drones and 3 kinds of medical packages are distributed, and then the DIjkstra algorithm is used to obtain the optimal distribution plan: San Juan Port is equipped with medical kits and three C-type drones, Arecibo Port allocates B-type drone patrol lines and medical kits for rescue, and Guayama Port only allocates drones for patrol lines. Using this distribution scheme greatly improves the efficiency of rescue, and distributes rescue materials reasonably and effectively, achieving the goals of optimal delivery path and optimal equipment utilization.

Glucosinolate (GSL) is associated with clubroot disease, which is caused by the obligate biotrophic protist Plasmodiophora brassicae. Due to the complicated composition of GSLs, their exact role in clubroot disease development remains unclear. By investigating clubroot disease resistance in cruciferous plants and characterizing the GSL content in seeds, we can determine if clubroot disease development is related to the components of GSLs. The difference in the infection process between Matthiola incana L. (resistant) and Brassica napus L. (susceptible) was determined. Root hair infection was definitely observed in both resistant and susceptible hosts, but no infection was observed during the cortical infection stage in resistant roots; this finding was verified by molecular detection of P. brassicae via PCR amplification at various times after inoculation. Based on the time course detection of the contents and compositions of GSLs after P. brassicae inoculation, susceptible roots exhibited increased accumulation of aliphatic, indolic, and aromatic GSLs in B. napus, but only aromatic GSLs were significantly increased in M. incana. Gluconapin, which was the main aliphatic GSL in B. napus and present only in B. napus, was significantly increased during the secondary infection stage. Quantification of the internal jasmonic acid (JA) concentration showed that both resistant and susceptible plants exhibited an enhanced level of JA, particularly in susceptible roots. The exogenous JA treatment induced aliphatic GSLs in B. napus and aromatic GSLs in M. incana. JA-induced aromatic GSLs may be involved in the defense against P. brassicae, whereas aliphatic GSLs induced by JA in B. napus likely play a role during the secondary infection stage. Three candidate MYB28 genes regulate the content of aliphatic GSLs identified in B. napus; one such gene was BnMYB28.1, which was significantly increased following both the treatment with exogenous JA and P. brassicae inoculation. In summary, the increased content of JA during the secondary infection stage may induce the expression of BnMYB28.1, which caused the accumulation of aliphatic GSLs in clubroot disease development.

Two double-stranded RNA (dsRNA) mycoviruses were found in isolate QSP5 of the rice blast fungus Magnaporthe oryzae. Sequence analysis of the two dsRNA mycoviruses revealed that one is closely related to Magnaporthe oryzae virus 2 (MoV2), and the other one is related to Magnaporthe oryzae chrysovirus 1-A (MoCV1-A). Therefore, they were named Magnaporthe oryzae virus 3 (MoV3) and Magnaporthe oryzae chrysovirus 1-C (MoCV1-C), respectively. In this paper, the molecular and structural characteristics of MoV3 were analyzed in detail. The full genome sequence (5181 bp) of MoV3 was obtained by cDNA cloning. Sequence analysis indicated that MoV3 has two overlapping open reading frames (ORF1 and ORF2). The 5'-proximal ORF1 encodes a putative coat protein (CP) with a molecular weight of 80,939 Da; the 3'-proximal ORF2 encodes a putative RNA-dependent RNA polymerase (RdRp) with a molecular weight of 90,506 Da. The stop codon of ORF1 overlaps the start codon of ORF2, with the tetranucleotide sequence AUGA, which is characteristic of members of the genus Victorivirus of the family Totiviridae. Phylogenetic analysis of RdRp and CP further supported the view that MoV3, a novel mycovirus, belongs to the genus Victorivirus of the family Totiviridae.

A double-stranded RNA (dsRNA) segment was isolated from a hypovirulent strain, HC025, of Sclerotinia sclerotiorum. The complete nucleotide sequence of the dsRNA was determined to be 2530 bp in length. Using the fungal mitochondrial genetic code, the positive strand of the dsRNA was found to contain a single large open reading frame (ORF) with the characteristic conserved motifs of the RNA-dependent RNA polymerase (RdRp). BLAST analysis revealed that RdRp shares 74% sequence identity with Sclerotinia sclerotiorum mitovirus 1 (SsMV1/KL-1). The positive strand of the dsRNA could be folded into potentially stable stem-loop structures at both the 5' and 3' terminal sequences. Moreover, the 5' and 3' terminal sequences were inverted complementary sequences and formed a panhandle structure. These results reveal that this dsRNA segment represents the replicative form of a mitovirus that is a strain of SsMV1 from the genus Mitovirus in the family Narnaviridae and was tentatively designated as Sclerotinia sclerotiorum mitovirus 1 (SsMV1/HC025). Sequence comparison and phylogenetic analysis suggest that mitovirus RdRp gene was evolutionarily related to plant mitochondrial genome. Our results demonstrate that SsMV1/HC025 infection exerted obvious effects on host biological properties. Hypovirulence feature and SsMV1/HC025 could be co-transmitted from hypovirulent strains to other virulent strains via hyphal contact. Thus, SsMV1/HC025 related to plant mitochondrial gene confers hypovirulence on S. sclerotiorum.

The present study investigated the protein expression levels of cluster of differentiation (CD)95, caspase-8, caspase−3 and poly(ADP-ribose) polymerase 1 (PARP1) in liver cancer and its association with clinical pathological parameters. The results demonstrated that the expression of CD95 correlated with histological differentiation, liver cirrhosis, lymph node metastasis and distant metastasis (P0.05). The expression of CD95 was upregulated using a plasmid, which led to an increase in the expression levels of caspase-8 and caspase-3 and a decrease in the expression of PARP1. Upregulation of CD95 also promoted the apoptosis of the liver cancer cells. These results indicated that CD95 was associated with liver cancer and promoted the apoptosis of liver cancer cells by caspase-8, caspase-3 and PARP1.

Estrogen receptor (ER)‑α36, a novel isoform of ER, primarily mediates non‑classical estrogen signaling. It has been reported that ER‑α36‑mediated growth stimulating signals are involved in the malignancy of gastric tumor cells. However, the mechanism underlying the regulation of ER‑α36 function in development of gastric cancer remains to be elucidated. The present study investigated the role of 78 kDa glucose‑regulated protein (GRP78) in the regulation of ER‑α36 expression and signaling during the growth of gastric tumor cells. It was demonstrated that GRP78 expression was detectable in gastric cancer tumor tissues, and was positively‑correlated with tumor stage, lymphatic metastasis and ER‑α36 expression (P

A hyperglycemic and hyperinsulinemic environment characteristic of type 2 diabetes causes insulin resistance. In adipocytes, defects in both insulin sensitivity and maximum response of glucose transport have been demonstrated. To investigate the molecular mechanisms, freshly isolated rat adipocytes were incubated in control (5.6 mM glucose, no insulin) and high glucose (20 mM)/high insulin (100 nM) (HG/HI) for 18 hours to induce insulin resistance. Insulin-resistant adipocytes manifested decreased sensitivity of glucose uptake associated with defects in insulin receptor substrate (IRS)-1 Tyr phosphorylation, association of p85 subunit of phosphatidylinositol-3-kinase, Akt Ser473 and Thr308 phosphorylation, accompanied by impaired glucose transporter 4 translocation. In contrast, protein kinase C (PKC)-ζ activity was augmented by chronic HG/HI. Inhibition of PKC-ζ with a specific cell-permeable peptide reversed the signaling defects and insulin sensitivity of glucose uptake. Transfection of dominant-negative, kinase-inactive PKC-ζ blocked insulin resistance, whereas constitutively active PKC-ζ recapitulated the defects. The HG/HI incubation was associated with stimulation of IRS-1 Ser318 and Akt Thr34 phosphorylation, targets of PKC-ζ. Transfection of IRS-1 S318A and Akt T34A each partially corrected insulin signaling, whereas combined transfection of both completely normalized insulin signaling. In vivo hyperglycemia/hyperinsulinemia in rats for 48 hours similarly resulted in activation of PKC-ζ and increased phosphorylation of IRS-1 Ser318 and Akt Thr34. These data indicate that impairment of insulin signaling by chronic HG/HI is mediated by dual defects at IRS-1 and Akt mediated by PKC-ζ.

Accumulating evidence indicates that cancer stem cells (CSC) play important roles in breast cancer occurrence, recurrence and metastasis as well as resistance to therapy. However, the roles of breast cancer stem cells in antiestrogen resistance and the underlying molecular mechanisms have not been well established. Previously, we identified and cloned a novel variant of estrogen receptor α, ER-α36, with a molecular weight of 36kDa. ER-α36 mediates rapid antiestrogen signaling and is highly expressed in ER-positive breast cancer stem/progenitor cells. In this study, we investigated the function and the underlying mechanism of ER-α36-mediated antiestrogen signaling in ER-positive breast cancer stem/progenitor cells. ER-positive breast cancer cells MCF7 and T47D as well as variants with different levels of ER-α36 expression were used. The effects of antiestrogens tamoxifen and ICI 182, 780 on breast CSC's ability of growth, self-renewal, differentiation and tumor seeding were examined using tumorsphere formation, flow cytometry, indirect immunofluorences and in vivo xenograft assays. The underlying mechanisms were also analyzed with Western blot analysis. We found that the cancer stem/progenitor cells enriched from ER-positive breast cancer cells were more resistant to antiestrogens than the bulk cells. Antiestrogens increased the percentages of the stem/progenitor cells from ER-positive breast cancer cell through stimulation of luminal epithelial lineage specific ER-positive breast cancer progenitor cells while failed to do so in the cells with knocked-down levels of ER-α36 expression. Our results thus indicated that ER-α36-mediated antiestrogen signaling such as the PI3K/AKT plays an important role in antiestrogen resistance of ER-positive breast cancer stem/progenitor cells.

To examine the expression patterns of ER-α36 and Cyclin D1 in human gastric cancer tissues and to investigate the effects of ER-α36-mediated estrogen signaling on the growth of gastric cancer cells, 117 samples of formalin-fixed and paraffin-embedded gastric cancer tumor tissues and 40 fresh gastric cancer tumor tissues were analyzed with immunohistochemistry assay and western blot analysis. ER-α36 expression was well correlated with gender (male:female ratio 2.88:1, P=0.01), invasion to serosa (P=0.01) as well as Cyclin D1 expression (P

Glucose-regulated protein 94 (GRP94) has been implicated in the promotion of tumor proliferation and metastasis. Previous studies have found that GRP94 is involved in the malignant growth of gastric carcinoma cells through estrogen receptor-α36 (ER-α36)-mediated estrogen signaling, but the underlying mechanism remains unclear. In the present study, we examined the expression levels of GRP94 and ER-α36 in tumor specimens from gastric cancer patients by immunohistochemistry, and found that both GRP94 and ER-α36 were highly expressed in the cytoplasms of gastric carcinoma cells. Furthermore, treatment with 17β-estradiol at a concentration of 10−12 M for 24 h increased the expression levels of GRP94 and ER-α36, and the phosphorylation levels of Akt at the Ser473 site (Ser473-Akt). In established SGC7901 gastric cancer cells with knockdown of ER-α36 expression, the levels of GRP94 and Ser473-Akt expression were significantly reduced. Thus, the Akt signaling pathway is a potentially important signaling pathway in ER-α36-GRP94-mediated gastric carcinogenesis.

Several previous studies have demonstrated that estrogen may protect cancer cells from endoplasmic reticulum stress-induced apoptosis. However, the molecular mechanisms involved are not fully understood. In the present study, human gastric adenocarcinoma SGC7901 cells were treated with tunicamycin (TM) to induce endoplasmic reticulum stress. This was demonstrated by increased glucose-regulated protein 78 expression and enhanced phosphorylation of protein kinase RNA-like endoplasmic reticulum kinase. Endoplasmic reticulum stress induced caspase-3-mediated apoptosis with the inhibition of Akt; the latter of which was measured by the activity-dependent phosphorylation at Ser473 of Akt. Simultaneous treatment of 10−9 M 17β-estradiol (E2) with TM may protect SGC7901 cells from endoplasmic reticulum stress-induced apoptosis by counteracting the inhibitory effect of TM on Akt, causing an increase in the phosphorylation of Ser473-Akt. It was concluded that low concentrations of E2 may counteract endoplasmic reticulum stress-induced inactivation of Akt to block caspase-3-mediated apoptosis.

Previously, a novel variant of estrogen receptor (ER)-α, ER-α36, was identified and cloned and reported to mainly mediate non-genomic estrogen signaling. More recently, we identified that ER-α36 is important for the invasion and lymph node metastasis of human gastric cancer. In the present study, the c-Src signaling pathway was demonstrated to be involved in the non-genomic estrogen signaling mediated by ER-α36 in SGC7901 gastric cancer cells. SGC7901 cells were subjected to the siRNA-mediated knockdown of ER-α36 (PLKO.1-PURO-SP6-ER-α36-L) or transfected with an ER-α36 upregulated expression plasmid (PLJM1-ER-α36-H) and treated with 17β-estradiol (E2β) and PP2, a c-Src protein inhibitor. The expression of ER-α36 and c-src/p-c-Src and cyclin D1 was examined by western blot analysis, and tumor cell growth was analyzed by cell proliferation and nude mouse xenograft assays. The ER variant, ER-α36, was shown to enhance gastric cancer cell proliferation through activation of the membrane-initiated c-Src signaling pathways, indicating that ER-α36 is important for the regulation of proliferation in gastric cancer. In addition, ER-α36 was shown to directly interact with c-Src by immunoprecipitation. The results of the present study indicate that the use of ER-α36 may be a targeted therapeutic approach in gastric cancer.

The aim of the present study was to examine the rate of BRAF mutation and the expression profiles of CK19, galectin-3, CD56, thyroid peroxidase (TPO) and Ki67 in papillary thyroid carcinoma (PTC) and papillary thyroid micro-carcinoma (PTMC). A total of 246 cases of thyroid disease were collected, including PTC, PTMC, nodular goiter (NG) and Hashimoto thyroiditis (HT). The results revealed that CK19 expression was 116/120 in PTC, 61/64 in PTMC, 2/34 in NG and 1/28 in HT. Galectin-3 positive expression was 115/120 in PTC, 60/64 in PTMC, 6/34 in NG and 4/28 in HT. TPO positive expression was 8/120 in PTC, 1/64 in PTMC, 30/34 in NG and 25/28 in HT. CD56-positive expression was 12/120 in PTC, 3/64 in PTMC, 33/34 in NG and 26/28 in HT. Ki67 labeling index was 2.52±0.46% in PTC (120 cases), 2.62±0.52% in PTMC (64 cases), 2.55±0.44% in NG (34 cases) and 2.58±0.48% in HT (28 cases). BRAF mutation rate was 93/120 in PTC, 47/64 in PTMC, 3/34 in NG and 2/28 in HT. These results suggested that expression patterns of CK19, galectin-3, CD56 and TPO and BRAF mutation exhibit diagnosis value in thyroid disease. However, Ki67-positive rate exhibits no notable diagnosis value in thyroid disease.