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The HA:Tb/Zn synthesized by hydrothermal method synchronously achieved good antibacterial effect, better osteogenic activity, and stable fluorescence.

The plant allelochemical coumarin effectively inhibits the germination of Brassica parachinensis (B. parachinensis) seeds. Quantification of endogenous phytohormones showed that contents of abscisic acid (ABA), ABA glucose ester, gibberellin A20 (GA20), GA3, GA15, GA24, GA9 and GA4 were higher in germinating seeds than in seedlings. Moreover, the presence of coumarin significantly reduced the content of bioactive GA4 which is thought to positively regulate seed germination. Histochemical staining and spectrophotometry of reactive oxygen species (ROS) revealed that exogenous GA3 and GA4+7 could effectively promote the production of endogenous ROS during germination and that the GA synthesis inhibitor paclobutrazol could effectively inhibit production of ROS. Coumarin significantly inhibited the accumulation of ROS, especially superoxide anion radical (${\rm O}_2^{{\cdot}{-}} $). This inhibitory effect could be restored by the addition of exogenous GA3 and GA4+7. Coumarin also inhibited the activity of the ROS-degrading enzymes such as superoxide dismutase, catalase and peroxidase as well as β-amylase in seeds and seedlings. Taken together, we propose a model for the regulation of seed germination in B. parachinensis by coumarin, Gas and ROS, in which coumarin may delay seed germination by reducing endogenous GA4, thus decreasing the accumulation of ROS.

Objectives To explore the potential molecular mechanism underlying the effect of green tea extract (TE), rich in tea polyphenols (TPs), on improving alcohol-induced liver injury. Methods Mice were intragastrically treated with 50% (v/v) alcohol administration (15 ml/kg BW) with or without three doses of TE (50, 120 and 300 mg TPs/kg BW) daily for 4 weeks, and biological changes were tested. Key findings The TE improved the functional and histological situations in the liver of the mice accepted alcohol administration, including enzymes for alcohol metabolism, oxidative stress and lipid accumulation. Interestingly, the TE increased the nuclear translocation of nuclear factor erythroid-2-related factor 2 (Nrf2), with the decreasing expression of kelch-like ECH-associated protein 1 (Keap1), indicating the association between the effect of TE with Nrf2-mediated antioxidant signalling. Moreover, the TE restored the activity of autophagy, showing as lifted Beclin-1 expression, LC3B-II/LC3B-I ratio, and decreased p62 expression. Importantly, all these effects were dose-dependent. Conclusions These findings provide a new notion for the first time that the TE preventing against alcohol-induced liver injury is closely related to accelerated metabolism of alcohol and relieved oxidative stress, which is associated with Nrf2 signalling activation and autophagy restoration, thus the reduction of lipid accumulation in liver.

The disease caused by Riemerella anatipestifer (R. anatipestifer) causes large economic losses to the global duck industry every year. Serotype-related genomic variation (such as in O-antigen and capsular polysaccharide gene clusters) has been widely used for the serotyping in many gram-negative bacteria. To date, there have been few studies focused on genetic basis of serotypes in R. anatipestifer. Here, we used pan-genome-wide association studies (Pan-GWAS) to identify the serotype-specific genetic loci of 38 R. anatipestifers strain. Analyses of the loci of 11 serotypes showed that the loci could be well mapped with the serotypes of the corresponding strains. We constructed the knockout strain for the wzy gene at the locus, and the results showed that the mutant lost the agglutination characteristics to positive antisera. Based on the of Pan-GWAS results, we developed a multiple PCR method to identify serotypes 1, 2, and 11 of R. anatipestifer. Our study provides a precedent for systematically analysing the genetic basis of the R anatipestifer serotypes and establishing a complete serotyping system in the future.HighlightsR. anatipestifer serotype-specific locus was identified by Pan-GWAS for the first time.Molecular serotyping multiplex PCR was developed based on O-antigen biosynthesis gene clusters

ProBiotic-4 is a probiotic preparation composed of Bifidobacterium lactis, Lactobacillus casei, Bifidobacterium bifidum, and Lactobacillus acidophilus. This study aims to investigate the effects of ProBiotic-4 on the microbiota–gut–brain axis and cognitive deficits, and to explore the underlying molecular mechanism using senescence-accelerated mouse prone 8 (SAMP8) mice. ProBiotic-4 was orally administered to 9-month-old SAMP8 mice for 12 weeks. We observed that ProBiotic-4 significantly improved the memory deficits, cerebral neuronal and synaptic injuries, glial activation, and microbiota composition in the feces and brains of aged SAMP8 mice. ProBiotic-4 substantially attenuated aging-related disruption of the intestinal barrier and blood–brain barrier, decreased interleukin-6 and tumor necrosis factor-α at both mRNA and protein levels, reduced plasma and cerebral lipopolysaccharide (LPS) concentration, toll-like receptor 4 (TLR4) expression, and nuclear factor-κB (NF-κB) nuclear translocation in the brain. In addition, not only did ProBiotic-4 significantly decreased the levels of γ-H2AX, 8-hydroxydesoxyguanosine, and retinoic-acid-inducible gene-I (RIG-I), it also abrogated RIG-I multimerization in the brain. These findings suggest that targeting gut microbiota with probiotics may have a therapeutic potential for the deficits of the microbiota–gut–brain axis and cognitive function in aging, and that its mechanism is associated with inhibition of both TLR4-and RIG-I-mediated NF-κB signaling pathway and inflammatory responses., Graphical abstract Image 1Long-term oral administration with ProBiotic-4, a complex probiotic preparation, could significantly improve the deficits of the microbiota–gut–brain axis and cognitive function in aged SAMP8 mice. The underlying mechanism is associated with inhibition of both TLR4-and RIG-I-mediated NF-κB signaling pathway and inflammatory responses.

Coronary heart disease (CHD) refers to a disease where coronary atherosclerosis induces stenosis or obstruction of the blood vessels. Endothelial progenitor cells (EPCs) function to protect and repair the vascular endothelium, and their functional activity state reflects the ability of the body to repair vascular damage. In the peripheral blood of patients with CHD, the density of EPCs decreases, and the function of EPCs is low.This study aimed to investigate the effects of a China Food and Drug Administration (CFDA)-approved prescription medicine, Tongxin, on the density and function of endothelial progenitor cells (EPCs) in peripheral blood.In this study, a randomized, single blind, parallel controlled clinical trial was used. The single blind subjects were subjects.The study took place in the Cardiology and Emergency Departments at Shanghai Municipal Hospital of Traditional Chinese Medicine in Shanghai, China.Participants were 48 patients with coronary heart disease at the hospital.Participants were randomly divided into 2 groups (n = 24 each): a control group and an intervention group. Both groups received routine drug treatments, such as platelet inhibitors, nitrates, β-receptor blockers, statins, angiotensin-converting-enzyme (ACE) inhibitors, angiotensin II receptor antagonists (ARBs), and calcium blockers. The control group was treated with the Shexiang Baoxin Pill, while the intervention group was treated with prescription Tongxin. The course of treatment was 3 months for both groups.Changes in the density and function of EPCs in the peripheral blood of the 2 groups were measured at baseline and postintervention, and the clinical efficacy of the 2 treatments was statistically analyzed.The density of EPCs was significantly higher in both groups after 3 months of treatment, compared to the densities at baseline (P.05). The change in density between baseline and postintervention was significantly greater for the intervention group than for the control group (P.05). For the control group, the proliferative vitality [optical density (OD)] value of the EPCs was significantly higher than that at baseline from the fourth day of treatment (P.05). In the intervention group, the OD value was significantly higher than that at baseline from the first day of treatment (P.05). Furthermore, the intervention group's cells began to enter the logarithmic growth phase of increase from the fifth day of treatment, and the group's increase as significantly higher than the control group's from the fifth to the seventh dayof treatment (P.05 for all 3 days). Moreover, the total effective rate was higher in the intervention group than in the control group (P.05).Prescription Tongxin can stimulate the release of EPCs from the bone marrow to the peripheral blood of patients with CHD, can significantly increase the proliferation of EPCs in the peripheral blood, and can improve the clinical symptoms of patients. Its curative effect was greater than that of the control treatment.

Epidemiological studies suggest that chronic periodontitis (CP) is closely associated with the incidence and progression of cognitive impairment. The present study investigated the causal relationship between CP and cognitive decline and the underlying mechanism in mice. Long-term ligature around the left second maxillary molar tooth was used to induce CP in mice. Severe alveolar bone loss and inflammatory changes were observed in gingival tissues, accompanied by progressive cognitive deficits during a 12-month period. We also observed cerebral neuronal and synaptic injury and glial activation in this mouse model of CP. Furthermore, CP mice exhibited significant dysbiosis of the oral and gut microbiota, disruption of the intestinal barrier and blood-brain barrier, increases in the serum contents of proinflammatory cytokines and lipopolysaccharide (LPS), and increases in brain LPS levels, Toll-like receptor 4 (TLR4) expression, nuclear factor-κB (NF-κB) nuclear translocation and proinflammatory cytokine mRNA levels. These results indicate that CP may directly induce progressive cognitive decline and its mechanism is probably related to microbiota-gut-brain axis disorders, LPS/TLR4/NF-κB signaling activation and neuroinflammatory responses in mice. Therefore, the microbiota-gut-brain axis may provide the potential strategy for the prevention and treatment of CP-associated cognitive impairment.

To investigate the effects of different stimultors (PHA, PMA and IL-2) and culture systems (PBMC and whole blood) on the proliferation of human peripheral blood lymphocyte subsets, so as to provide the experimental basis for selecting the appropriate system according to the experimental purposes.A total of 10 ml serum samples were collected from healthy volunteers (n=6). The 300 μl whole blood was directly used to detect lymphocyte subsets by flow cytometry. The 400 μl whole blood were inoculated respectively with 3 different stimulators at 37℃ and 5% CO2 for 60 h; Three different stimulators were also added to the PBMC which were isolated from 2 ml whole blood. Then the proliferation ability of lymphocyte subsets was analyzed by flow cytometry.After the PBMC were stimulated with PHA, CD4Lymphocyte proliferation stimulated by PMA is the fastest, while the effect of IL-2 on the lymphocyte subset proportion stimulated by IL-2 is the minimal. After being stimulated by PHA the division cycles of lymphocyte are the most.

Age-related cognitive decline is associated with chronic low grade neuroinflammation that may result from a complex interplay among many factors, such as bidirectional communication between the central nervous system (CNS) and gut microbiota. The present study used 2-month-old (young group) and 15-month-old (aged group) male C57BL/6 mice to explore the potential association between age-related cognitive decline and the microbiota-gut-brain axis disorder. We observed that aged mice exhibited significant deficits in learning and memory, neuronal and synaptic function compared with young mice. Aged mice also exhibited significant dysbiosis of the gut microbiota. Disruptions of the intestinal barrier and blood-brain barrier were also observed, including increases in intestinal, low-grade systemic and cerebral inflammation. Furthermore, plasma and brain levels of lipopolysaccharide (LPS) were significantly higher in aged mice compared with young mice, with increasing expression of Toll-like receptor 4 (TLR4) and myeloid differential protein-88 (MyD88) and the nuclear translocation of nuclear factor κB (NF-κB) in intestinal and brain tissues. These findings showed that microbiota-gut-brain axis dysfunction that occurs through LPS-induced activation of the TLR4/NF-κB signaling pathway is implicated in age-related neuroinflammation and cognitive decline.

The dual-layer Li2TiO3 coating of LiNi0.5Co0.2Mn0.3O2 (NCM523) compound has been successfully synthesized by a facile method for the first time. The EDS results show that the dual-layer coating is more uniform and smoother than the single-layer coating. Electrochemical tests show that the double-layer 0.5 mol % Li2TiO3-coated NCM can deliver an initial discharge capacity of 180 mAh g−1 at 0.1 C in the voltage range of 2.8–4.3 V, with capacity retention of 94.2 % after 80 cycles at 0.5 C, while those of the pristine one are only 172.1 mAh g−1and 87 %, respectively. The enhanced electrochemical performance is attributed to the stable dual-layer Li2TiO3 that serves as a 3D-diffusion pathway for Li+ ions and a protective layer that prevents side reactions between electrode and electrolyte. Thus, the double-layer coating by Li2TiO3 is a promising method to improve rate capability and cycle performance of NCM523.

Background Individuals with sub-syndromal depression (SSD) are at increased risk of incident depressive disorders; however, the ideal therapeutic approach to SSD remains unknown. Objective To evaluate the effects of electroacupuncture (EA) and cognitive behavioural therapy (CBT), alone or in combination, on depressive symptoms. Methods Undergraduate students with SSD were recruited and allocated to one of four groups based on their preferences: EA (n=6), CBT (n=10), EA+CBT (n=6), and untreated control (n=11) groups. Six weeks of treatment were provided in the first three groups. Clinical outcomes were measured using the 17-item Hamilton Depression (HAMD-17) rating scale, Center for Epidemiologic Depression (CES-D) scale, WHO Quality of Life-Brief version (WHOQOL-BREF) questionnaire, and clinical remission rate. Results All 33 subjects were included in an intent-to-treat analysis. Statistically significant improvements in HAMD-17, CES-D, and WHOQOL-BREF scores and a higher remission rate were found in the EA, CBT, and EA+CBT intervention groups compared with the control group (all pConclusions Early intervention may alleviate depressive symptoms in SSD. EA and CBT may have differential effects on certain symptoms. Combination therapy targeting both physical and psychological symptoms may represent an ideal strategy for SSD intervention. However, randomised trials with larger sample sizes are needed. Trial Registration Number ChiCTR-TRC-10000889; Results.

Migraine is a disabling neurovascular disorder, which increases risk of cardiovascular events and is a social burden worldwide. The present first-line anti-migraine medications can cause overwhelming side-effects, of which one includes the onset of cardiovascular disease. As one of the marketed Tibetan drugs, Ru-yi-Zhen-bao Pills (RYZBP) have been clinically used to treat cardiovascular disorders and as anti-migraine medication. However, there is currently no research exploring the anti-migraine actions of RYZBP.The current research was designed to assess the anti-migraine roles of RYZBP and explore the underlying mechanisms in a nitroglycerin (NTG)-induced migraine rat model trial.120 rats were randomly divided into the following six groups of 20 rats each: normal control group, model control group, positive control group, and RYZBP high/medium/low-dose groups (Ru-yi-Zhen-bao Pills; TH 1.00 g/kg, TM 0.50 g/kg and TL 0.25 g/kg). All rats were administered intragastrically for 7 consecutive days, which were subcutaneously injected with the NTG (10 mg/kg) after the last gavage (except in the normal control group). 3min after NTG treatment, 30 rats (5 rats from each group) were anesthetized and devoted to electroencephalogram(EEG) testing, which was used to evaluate the analgesic effect of RYZBP. One hour after NTG treatment, the rest of the 90 rats (15 rats from each group) were anesthetized and midbrain tissue sample was dissected. The dissection was then washed with physiological saline and collected. The histopathological changes in the periaqueductal gray(PAG) of 5 tissue samples were determined by aematoxylin-eosin (HE) staining, as well as an estimation of substance P (SP) and neurokinin 1 receptor (NK1R) expression through immunohistochemically staining(IHC). Another 5 midbrain preparations were carried out to evaluate calcitonin gene-related peptide (CGRP), proenkephalin (PENK), SP, and cholecystokinin (CCK) expressions by real-time quantitative polymerase chain reaction (RT-qPCR). The rest of the 5 brainstem tissues were then used to measure CCK, CGRP, and opioid peptide receptor (DORR) levels by western blotting(WB).In the EEG test, RYZBP (TM 0.50 g / kg) treatment transformed the EEG pain-wave of the NTG-induced migraine model rats in different time period. In the mechanism assay, compared with the model control group, RYZBP pretreatment reduced inflammatory cell infiltration, fibrosis and vacuolation of neuronal cells of PAG tissue seen by HE staining. IHC experiments further showed that RYZBPTM up-regulated SP expression levels and enhanced NK1R levels in the NTG-induced migraine rats (P 0.05). Therapeutic administration of RYZBP also increased PENK mRNA expression and DORR protein level. Both RT-qPCR and western blotting trials indicated that RYZBP treatment significantly decreased CCK and CGRP expression levels (P 0.01 or P 0.05) in the NTG-induced migraine rats.RYZBP has the potential to be an effective anti-migraine treatment through suppressing the EEG pain-wave, increasing the levels of SP, PENK, DORR and reducing expression of CCK and CGRP. Mediating the PAG anti-nociceptive channel and inhibiting central sensitization were the two potential mechanisms, which offers further evidence for clinical therapy.

The experiment was conducted in the plots that had been enclosed for nearly 20 years in the hilly Loess Plateau region. The effects of trampling disturbance on the biological soil crust (biocrust) coverage, soil organic carbon (SOC), soil easily oxidizable carbon (SEOC), SOC mineralization amount and mineralization rate were investigated. The biocrust SOC mineralization potential after disturbance in different soil layers were simulated by a first-order kinetic equation. The results showed that the coverage of cyanobacteria biocrust and moss biocrust significantly decreased with the increases of disturbance intensity. Compared to no disturbance, the cyanobacteria coverage declined by 264%-339% and moss coverage declined by 46%-127%. Compared to no disturbance, SOC content in biocrust layer significantly decreased by 211%-300%. No significant difference was found among the five disturbance intensities. Disturbance increased SEOC content in biocrust layer, and the variation amounted to 1.5-3.4 g·kg以黄土丘陵区近20年的封禁地为研究对象,采用野外调查结合室内分析,研究了不同强度的踩踏干扰对半干旱生态系统下生物结皮盖度、有机碳、易氧化碳和有机碳矿化量、矿化速率的影响,并应用一级动力学方程模拟了踩踏干扰下各层次生物土壤结皮有机碳矿化潜力的变化特征.结果表明: 干扰后藻结皮和藓结皮盖度呈现逐渐降低的趋势,干扰度间差异显著;与不干扰处理相比,藻结皮盖度降幅在264%～339%,藓结皮盖度降幅在46%～127%.与不干扰处理相比,干扰显著降低了生物结皮层有机碳含量,降幅在211%～300%,干扰度间差异不显著;干扰增加了生物结皮层易氧化碳含量,增加量在1.5～3.4 g·kg

The clinical effect of electroacupuncture on depression is widely recognized. However, the signal transduction pathways and target proteins involved remain unclear. In the present study, rat models of chronic restraint stress were used to explore the mechanism by which electroacupuncture alleviates depression. Rats were randomly divided into control, model, and electroacupuncture groups. Chronic restraint stress was induced in the model and electroacupuncture groups by restraining rats for 28 days. In the electroacupuncture group, electroacupuncture pretreatment at Baihui (GV20) and Yintang (GV29) acupoints was performed daily (1 mA, 2 Hz, discontinuous wave, 20 minutes) prior to restraint for 28 days. Open field tests and body weight measurements were carried out to evaluate the depressive symptoms at specific time points. On day 28, the crossing number, rearing number, and body weights of the model group were significantly lower than those in the control group. Behavior test results indicated that rat models of depressive-like symptoms were successfully established by chronic restraint stress combined with solitary raising. On day 28, an isobaric tag for a relative and absolute quantitation-based quantitative proteomic approach was performed to identify differentially expressed proteins in hippocampal samples obtained from the model and electroacupuncture groups. The potential function of these differential proteins was predicted through the use of the Cluster of Orthologous Groups of proteins (COG) database. Twenty-seven differential proteins (uncharacteristic proteins expected) were selected from the model and electroacupuncture groups. In addition to unknown protein functions, COG are mainly concentrated in general prediction function, mechanism of signal transduction, amino acid transport and metabolism groups. This suggests that electroacupuncture improved depressive-like symptoms by regulating differential proteins, and most of these related proteins exist in nerve cells.

A complex of low-molecular cationic peptides, extracted from human urine by a combination membrane ultrafiltration and weak cation exchange chromatography, was characterized in this study. It provides a simpler solution for the development of novel antimicrobial peptides from biological liquid waste.

To analyze the cellular function of the newly discovered DNA damage repair factor WDR70, and investigate the mutation in ovarian cancer to verify if function loss of theTheWe found gene silencing ofOur results defined that

Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive CD8+ T cells with induction of IFN-γ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-γ levels were measured by ELISA and flow cytometry, respectively. CD8+ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-γ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of CD8+ T cells from tumor bearing mice, while anti-IFN-γ blocked the function of CD8+ T cells, suggesting that IFN-γ mediated the cytolytic activity of CD8+ T cells. Furthermore, in vivo neutralization of CD8+ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating CD8+ T cells and stimulating them to secrete IFN-γ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.

Objective: Twist, a basic helix-loop-helix transcription factor, plays a key role in the metastatic progression of human cancer. Matrix metalloproteinase (MMP)-9 is an endopeptidase that digests basement membrane type IV collagen, therefore being possibly related to tumor progression. It has been reported that Twist and matrix metalloproteinase-9 (MMP-9) are expressed in gastric cancers. However, the exact roles of Twist and MMP-9 in tumor metastasis and prognosis remain unclear. The aim of this study was to casts light on this question. Methods:Twist and MMP-9 expression in tissue sections of 37 gastric carcinomas was evaluated with immunohistochemistry. The staining results were compared with clinicopatholgic features and to patients’outcome. Results: Twist positive expression was significantly increased in gastric cancer cases with lymph node metastasis (P=0.023). But no correlations were found between MMP-9 overexpression and clinicopathologic features, such as recurrence, TNM stage, and lymph node metastasis. Overall survival (OS) was significantly correlated with recurrence, serosa invasion, TNM stages, distant metastasis, and MMP-9 (P=0.027, 0.021, 0.000, 0.024 and 0.036, respectively). Disease-free survival (DFS) was prominently related to recurrence location, serosa invasion and TNM stages (P=0.000, 0.038 and 0.003, respectively). In the Cox regression multivariate analysis, TNM stage, distant metastasis and MMP-9 were significantly associated with prognosis of gastric cancer ( P=0.002, 0.019, and 0.032, respectively). Conclusions: This study showed Twist positive expression to be significantly correlated with lymph node metastasis in gastric cancer. MMP-9 overexpression is associated with OS, suggesting that MMP-9 is a prognostic indicator for survival in patients with gastric cancer.

Many toxic and undegradation organic pollutants to people health are existed in explosive wastewater. The conventional treatments are unable to effectively remove them. This paper treated the explosive wastewater of TNT by using a self-made discontinuity reactor, Supercritical Water Oxidation (SCWO) system. The experimental results show that SCWO is a high-efficiency, clean treatment and disposal technique for TNT explosive wastewater. Temperature and reaction time serve as the main influence factors in removing COD of wastewater, with which add, the COD removal rate obviously increases. Under no oxygen condition, partially oxidation existed in SCWO.This shows that supercritical water is a well solvent and has oxidation. At 550°C, 24MPa, and 120s and at oxygen excess 300%, the COD removal rate of wastewater is over 99% and its main toxic pollutants TNT are thoroughly disposed. The discharge standard of environmental protection is met.

Background Riemerella anatipestifer infection is a contagious disease that has resulted in major economic losses in the duck industry worldwide. This study attempted to characterize CRISPR-Cas systems in the disease-causing agent, Riemerella anatipestifer (R. anatipestifer). The CRISPR-Cas system provides adaptive immunity against foreign genetic elements in prokaryotes and CRISPR-cas loci extensively exist in the genomes of archaea and bacteria. However, the structure characteristics of R. anatipestifer CRISPR-Cas systems remains to be elucidated due to the limited availability of genomic data. Results To identify the structure and components associated with CRISPR-Cas systems in R. anatipestifer, we performed comparative genomic analysis of CRISPR-Cas systems in 25 R. anatipestifer strains using high-throughput sequencing. The results showed that most of the R. anatipestifer strains (20/25) that were analyzed have two CRISPR loci (CRISPR1 and CRISPR2). CRISPR1 was shown to be flanked on one side by cas genes, while CRISPR2 was designated as an orphan. The other analyzed strains harbored only one locus, either CRISPR1 or CRISPR2. The length and content of consensus direct repeat sequences, as well as the length of spacer sequences associated with the two loci, differed from each other. Only three cas genes (cas1, cas2 and cas9) were located upstream of CRISPR1. CRISPR1 was also shown to be flanked by a 107 bp-long putative leader sequence and a 16 nt-long anti-repeat sequence. Combined with analysis of spacer organization similarity and phylogenetic tree of the R. anatipestifer strains, CRISPR arrays can be divided into different subgroups. The diversity of spacer organization was observed in the same subgroup. In general, spacer organization in CRISPR1 was more divergent than that in CRISPR2. Additionally, only 8 % of spacers (13/153) were homologous with phage or plasmid sequences. The cas operon flanking CRISPR1 was observed to be relatively conserved based on multiple sequence alignments of Cas amino acid sequences. The phylogenetic analysis associated with Cas9 showed Cas9 sequence from R. anatipestifer was closely related to that of Bacteroides fragilis and formed part of the subtype II-C subcluster. Conclusions Our data revealed for the first time the structural features of R. anatipestifer CRISPR-Cas systems. The illumination of structural features of CRISPR-Cas system may assist in studying the specific mechanism associated with CRISPR-mediated adaptive immunity and other biological functions in R. anatipestifer. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-3040-4) contains supplementary material, which is available to authorized users.

Aniline monomer coated on the nanotubes in HCl solution was used to prepare a composite of multi-wall carbon nanotube/polyaniline (MWCNT/PANI) by in-situ polymerization. Multi-wall carbon nanotubes were treated by the mixed acid (HNO3:H2SO4in a ratio of 1:3). The content of MWCNT in the samples was 0-20 wt%. The nanocomposites were characterized by Thermogravimetric analysis (TGA), Fourier transform-infrared spectroscopy (FT-IR), Raman spectroscopy and scanning electron microscopy. The mechanical properties of the MWCNT/PANI nanocomposites were also measured. The results showed that the MWCNT was well dispersed in water after purification. Both FT-IR and Raman spectra illustrated the presence of MWCNT in the composites; the interaction between PANI and MWCNT was proved. The Young΄s modulus was 618MPa with 20wt% loading. The effects of treatment of MWCNT on the mechanical properties of MWCNT/PANI nanocomposites are discussed.

Objective To investigate the physiological features of patients with psoriasis vulgaris during sleep.Methods Thirty-six psoriasis patients were classified into the group with "blood-heat" syndrome(n=21) and the group with "blood-dry" syndrome(n=15).Fifteen healthy volunteers served as control.All the subjects underwent a nocturnal sleep examination using the micro-movement sensitive mattress sleep monitoring system(MSMSMS).The sleep indices and nocturnal heart rate dynamics of the patients were compared with that of the control,and also between the two groups with different syndrome.Results In comparison with the control group,both psoriasis patient groups showed the phenomena of poor sleep quality,such as the shallow sleep phase,disorder of sleep rhythm and increased slight-arousals.The slight-arousal occurred more often in blood-heat syndrome group than in blood-dry syndrome group,while the sleep latency was elongated more evidently in blood-dry syndrome group.Compared with the control group,the two patient groups presented a little decrease in heart rate(HR) after sleeping.However,the HR increased in the first phase of sleep,and the variation coefficient of HR increased in the whole sleep period.An increase in HR variation coefficient was more obvious in blood-heat syndrome group than in blood-dry syndrome group(P < 0.05).Conclusions The phenomena of decreased sleep quality and the variation of nocturnal HR in psoriasis patients may be due to the failure of Yin to control Yang,and disturbance of heat to the mind induced by blood-heat and Yin deficiency.The degrees of the phenomena are different between the two psoriasis groups.There is also a decrease in parasympathetic activity and relative increase in sympathetic activity in both psoriasis groups,with manifestation of different features in patients with different syndromes.

It has been well recognized that human epidermal growth factor receptor 2 (HER2) level in breast cancer (BC) is closely related to the malignant biologic behaviors of the tumor, including invasion and metastasis. Yet, there has been a lack of directly observable evidence to support such notion. Here we report a quantum dots (QDs)-based double-color imaging technique to simultaneously show the HER2 level on BC cells and the type IV collagen in the tumor matrix. In benign breast tumor, the type IV collagen was intact. With the increasing of HER2 expression level, there has been a progressive decrease in type IV collagen around the cancer nest. At HER2 (3+) expression level, there has virtually been a total destruction of type IV collagen. Moreover, HER2 (3+) BC cells also show direct invasion into the blood vessels. This novel imaging method provides direct observable evidence to support the theory that the HER2 expression level is directly related to BC invasion.

The objective of the study was to compare the clinical value of preoperative serum carbohydrate antigen 19-9 (CA19-9) and carbohydrate antigen 242 (CA242) in diagnosis and prognosis for 5-year recurrence-free survival (RFS) in patients with colorectal cancer (CRC). Preoperative serum CA19-9 and CA242 concentrations were detected by C12 protein chip diagnostic system in 185 patients with CRC, and informative data were collected during 5-year follow-up periods. The value of CA19-9 and CA242 in diagnosis and prognosis for 5-year RFS as well as their consistencies and correlations were comparatively analyzed. The sensitivities of CA19-9 and CA242 were only 19.5 and 20%, respectively; the efficiencies of two TMs were 53.9 and 54.2%, respectively; and two TMs increased significantly with advancing clinical stages (P < 0.0001). Preoperative CA19-9 and CA242 levels correlated with stage (r, 0.411 and 0.408) and CEA concentration (r, 0.553 and 0.630). The concentrations of two TMs closely correlated with each other (r = 0.829), and two TMs had a very strong consistency in diagnosis (κ = 0.931). Among 88 of 185 cases with complete follow-up information on RFS, patients with positive preoperative serum CA19-9 or CA242 had higher 5-year recurrent rates (72.2% vs. 44.3%, P = 0.034; 76.5% vs. 43.7%, P = 0.015) and reduced median RFS (14 vs. 36 months, 12 vs. 36 months) compared with those with negative TMs. The consistency of predicting prognosis for RFS of two TMs was extremely strong (κ = 0.964). ROC curves analysis showed that CA242 test performed better than CA19-9 test (AUC, 0.648 vs. 0.605). Univariate analysis showed that preoperative serum status of both TMs was correlated with 5-year RFS (P < 0.05), whereas multivariate Cox regression model analysis revealed that none of them were independent prognostic factors for RFS. Both CA19-9 and CA242 had strong consistencies in diagnosis and prognosis for predicting 5-year RFS. CA242 demonstrated superior value to CA19-9 in CRC.

Tumor growth and progression depends on their microenvironment, which undergoes constant co-evolution because of the dynamic tumor-stormal interactions. Such co-evolution has long been under appreciated due to the lack of appropriate technology platforms to simultaneously reveal these complex interactions. Here we report on a quantum dots based multiplexed imaging and spectrum analysis technology to simultaneously study major components of tumor stroma, including type IV collagen, tumor angiogenesis, macrophages infiltration and tissue destructive proteolytic enzyme matrix metalloproteinase 9. The new technology revealed a panoramic picture of the tempo-spatial co-evolution of tumor cells and their stroma at the architecture level. Four patterns of tumor invasion with distinctive co-evolution features were identified as Washing pattern, Ameba-like pattern, Polarity pattern and Linear pattern. This quantum dots based multiplexed technology could help gain new insight into the complex process of tumor invasion, and formulate new anti-cancer strategies.

The dynamic environments in riparian zones support a variety of life-history strategies, which constitute a fundamental mechanism for development and maintenance of biodiversity in riparian forest ecosystems. To demonstrate the effect of life-history strategies on biodiversity, we investigated community-level diversity and its relationship to environmental variability in the riparian Populus euphratica forests of the Ejina Oasis. Communities were divided into 14 associations on the basis of their species importance values. The Simpson’s index, Shannon-Wiener index, Pielou’s evenness index and the regional Whittaker’s index were applied to calculate community diversity. An ordination of the sample plots was carried out by correspondence analysis (CA). Biodiversity was relatively low across the entire study area, but there was high community diversity (β w = 8.09) due to the spatial heterogeneity of habitats in different plots. In addition, we investigated the relationship between biodiversity and several environmental factors, such as water availability, community components and soil conditions. We conclude that the heterogeneity of soil and water availability drives community diversity patterns in riparian zones and that community-level diversity favors the maintenance of species diversity in the P. euphratica forests in the Ejina Oasis.

To evaluate the prognostic value of preoperative serum carbohydrate antigen19-9 (CA19-9), carcinoembryonic antigen (CEA), and carbohydrate antigen 125 (CA125) for overall survival (OS) in patients with colorectal cancer (CRC). Preoperative serum CA19-9, CEA, and CA125 levels were detected by C12 protein chip diagnostic system in 80 patients with CRC, and the association of their preoperative status with the patients' OS was analyzed. Patients with positive preoperative serum CA19-9, CA125, and CEA had a worse survival comparing with those negative patients, the difference in CA19-9 and CA125 reached statistical significance (Log rank test, P0.05), but the difference of CEA did not achieve statistical significance (Log rank test, P0.05). The OS of patients with CRC with three positive tumor markers (TMs)1 or 2 positive TMsnegative TM, simultaneously positive serum CA19-9, CEA, and CA125 was the most dismal predicting factor for OS. Spearman correlations revealed that the preoperative serum status of CA19-9 (P=0.000), CEA (P=0.000), and CA125 (P=0.000) had correlations with stage; the preoperative serum CA125 status (r=-0.385, P=0.000) and stage (r=-0.457, P=0.000) and location (r=0.223, P=0.047) had correlations with OS. Multivariate analysis with Cox regression model showed that only stage (P=0.000) and the preoperative serum CA125 status (P=0.016) were independent prognostic factors for OS of patients with CRC. The preoperative status of serum CA19-9, CEA, and CA125 had correlations with CRC stage. The simultaneously positive serum CA19-9, CEA, and CA125 was the most dismal predicting factor for OS of patients with CRC. Preoperative serum CA125 status and stage were independent prognostic factors for OS of CRC.

As a cis element responsive to oxidative stresses, the promoter of amorphadiene synthase gene (pADS) from Artemisia annua was also found to be inducible by salicylic acid (SA) and methyl jasmonate (MJ), which had been evident by phenotyping pADS-driven β-glucuronidase reporter gene (GUS) in transgenic Nicotiana tabacum incubated with SA/MJ. Furthermore, SA/MJ-treated A. annua exhibited a correlation of the up-regulation of ADS gene with the emission of singlet oxygen (1O2), suggesting that SA/MJ induced artemisinin overproduction through invoking 1O2 burst. Supplement with Rose Bengal (RB), a photosensitizing 1O2 generator, or direct exposure to a 1O2-generating pool, however, failed to facilitate artemisinin accumulation in subjected A. annua. Alteration of metabolite profiles and fluctuation of H2O2 level suggested that RB treatment might have led to a diminished endogenous 1O2 release from A. annua, thereby reiterating an important role of the endogenous 1O2 rather than the exogenous 1O2 in enhanced artemisinin production.

To interfere with the endogenous gene encoding squalene synthase (SS) responsible for sterol biosynthesis, an antisense SS cDNA ( asSS ) was introduced into the genome of A. annua . Following the decline of SS mRNA and synchronous reduction of total sterols, mRNA coding for amorpha-4,11-diene synthase (ADS) tremendously elevated, whereas mRNAs for sesquiterpene cyclases encompassing 8-epicedrol synthase (EPS), β-caryophyllene synthase (CS) and β-farnesene synthase (FS) declined or kept homeostasis, suggesting a bias in redirection of the partially blocked metabolic flow to amorpha-4,11-diene. By phenotyping the chimeric ADS promoter-fused β-glucuronidase gene ( pADS-GUS ), overexpression of the pADS-GUS reporter was detected upon chilling exposure and ultraviolet (UV) irradiation. In transgenic plants that expose to chilling, ADS and EPS were dramatically up-regulated, but CS was down-regulated and FS and SS were unaffected, implying further chilling-induced diversion of carbon flux among sesquiterpenes. Accordingly, ADS and cytochrome P450 reductase (CPR) were also chilling-inducible, while cytochrome P450 monooxygenase (CYP71AV1) reflected a complicated chilling profile. A maximal artemisinin content was accounted for 16.6 mg/g or 3.7-fold increase in a chilled transgenic line. Fascinatingly, an extraordinarily high artemisinin content of 23.5 mg/g or 5.2-fold increase was measured after 15-month storage of the ground powder from the identical chilled transgenic tissue, demonstrating non-enzymatic conversion from a putative artemisinin precursor to artemisinin. Moreover, a relevance of chilling exposure with singlet oxygen ( 1 O 2 ) emission and artemisinin overproduction was established. These results indicated that the mevalonate pathway has been first skewed to amorpha-4, 11-diene and shunted ultimately to artemisinin through a synergy of genetic modification with environmental superinduction.

To investigate the dynamic fluctuation of terpenoid relevant transcriptomics in transgenic ARTEMISIA ANNUA plants that express the genomic integrated antisense squalene synthase gene ( ASSS), we have quantified the transcript levels of the sterol anabolic SS gene as well as artemisinin biogenetic amorphadiene synthase (ADS), cytochrome P450 monooxygenase (CYP71AV1) and cytochrome P450 reductase (CPR) genes by real-time fluorescent quantitative polymerase chain reaction (RFQ-PCR). The SS mRNA level in transgenic plants sharply droped to 7.4 % - 55.3 % (i. e., 44.7 - 92.6 % reduction as the wild-type control), strongly implying that the expression of endogenous SS gene is significantly suppressed by the exogenous ASSS gene. In a synchronous fashion, ADS, CYP71AV1 and CPR mRNA levels elevated with the decline of SS mRNA level in transgenic plants, and the maximal ADS, CYP71AV1 and CPR mRNA levels in transgenic plants were 3.0-, 4.4- and 2.5-fold, respectively, higher than those in the control. Without a lethal effect but with a distinguishable impact on the organogenesis and morphology of transgenic plants, the down-regulation of SS gene has also led to the coordinated overexpression of ADS, CYP71AV1 and CPR genes together with the overproduction of artemisinin although no fully perfect correlation among the available experimental data has been shown.

To elucidate the fine-tuned temporal and spatial modulation of artemisinin production in annual wormwood (Artemisia annua), we conducted enzyme-linked immunosorbent assay-based immunoquantification of three key enzymes involved in artemisinin biosynthesis, amorpha-4,11-diene synthase (ADS), cytochrome P450 monooxygenase (CYP71AV1), and cytochrome P450 reductase (CPR), in various tissues and under different growth conditions. The field-grown plants accumulate abundant ADS and CYP71AV1 but a trace amount of CPR in all tested tissues. Furthermore, ADS and CYP71AV1 accumulations in leaves are 16- and eightfold higher than in roots, and ten- and fourfold higher than in stems, respectively, demonstrating a tissue-specific expression pattern. Interestingly, the flowering field plants and cold-acclimated cultural plants produce higher levels of ADS and CYP71AV1 than non-flowering field plants or untreated cultural plants, indicating the environmental and developmental induction on ADS and CYP71AV1 genes and providing possible explanation for the observation that elevation of artemisinin level occurs after flowering.

To isolate and verify novel genes from qinghao (Artemisia annua) based on the development-specific and environment-induced transcriptomics, leaves have been harvested from the flowering A. annua plants and exposed to low temperature for isolation of total RNAs and cloning of full-length cDNAs and cDNA fragments, or expressed sequence tags (ESTs). After being sequenced and browsed for homology, these sequences have been submitted to GenBank. Among the accessed 75 sequences, 4 full-length cDNAs are highly homologous to the known A. annua genes, but 71 ESTs are absent in the sequence records of A. annua genes, in which 34 sequences are homologous to other plant genes, including 24 identified protein-coding sequences and 10 unidentified protein-coding sequences, while other 37 sequences are not present in the sequence records of any plant genes, representing the first cloned plant genes. In order to investigate the responsive patterns of A. annua genes to extreme environmental stresses, especially low temperature, the expression levels of 3 critical qinhaosu (artemisinin) biosynthetic genes, ADS, CYP71AV1 and CPR, have been measured in pre-and post-chilling A. annua seedlings cultured in vitro by semi-quantitative PCR (SQ-PCR). Consequently, ADS and CYP71AV1 genes are strongly induced by chilling, but CPR gene is not significantly affected by such treatment. Furthermore, induction of these genes by chilling can be potently suppressed by Ca2+ channel inhibitor LaCl3 or Ca2+ chelator EGTA, suggesting a putative involvement of Ca2+-CaM signal transduction pathway in chilling-induced overexpression of ADS and CYP71AV1 genes. The real-time fluorescent quantitative PCR (RFQ-PCR) assay of A. annua seedlings exposed to chilling has shown that the expression level of CaM gene is up-regulated for more than 2.5 folds, thereby confirming our above inference on the relevance of Ca2+-CaM-mediated signal transduction to chilling-induced gene overexpression. Finally, 7 newly isolated A. annua ESTs have been functionally annotated by RFQ-PCR, which indicates that the chilling stress-induced overexpression levels of D/LTSRP, UBE, AR/DAP and POD1 genes are augmented approximately for 8, 5, 2.3, and 1.5 folds, respectively, but those of CHI and RGP genes are not predominantly up-or down-regulated. The present study has preliminarily explored the chilling-induced overexpression patterns of 3 artemisinin biosynthetic genes and 7 novel A. annua ESTs at the transcriptional level, which should further facilitate our understanding of the intrinsic rule and mechanism underlying the coordinative regulation manner of artemisinin biosynthesis and accumulation, and yield substantial insight into improvement of A. annua by the metabolic engineering-guided breeding strategy.

The potential of malaria parasite (Plasmodium falciparum) generating resistance to currently used antimalarial drugs is a major scourge for malaria patients. The multiple targets on which antimalarial drugs act hinder the occurrence of such resistance. As an essential metabolic enzyme involved in energy production in the parasite, the lactate dehydrogenase of P. falciparum (PfLDH) provides an alternative target for parasite killing. In the present study, the PfLDH gene was amplified from the FCC1/HN isolate of PfLDH and functionally expressed in Escherichia coli, which had been subsequently identified by Western blotting with the recombinant PfLDH protein-raised rabbit polyclonal antiserum, and further confirmed by the hybridization with lytic suspension of cultured parasites. The recombinant PfLDH protein was purified for uniform measurement of enzymatic activity and immobilized on 96-well plates for detection of PfLDH-targeted inhibitors. Hematin (HT) and chloroquine (CQ), two well-known inhibitors of PfLDH, were chosen for evaluating the feasibility of the in vitro colorimetric enzyme assay. The results showed that the enzymatic activity of PfLDH sharply dropped in a concentration-dependent pattern, as HT was involved, but only slightly dropped in the presence of CQ. Furthermore, this enzyme assay was applied to screening inhibitory component (s) of PfLDH from the crude extracts of medicinal plants, Pueraria lobata, Amomum villosum, Dichroa febrifuga, and Polygonum cuspidatum, in which the suppression of malaria growth was previously observed for P. lobata and D. febrifuga. Consequently, the correlation of enzyme inhibition with growth suppression was observed for the crude extracts of P. lobata although further investigations should be pursued for elucidation of the exact compound exhibiting inhibitory effect to PfLDH. The present recombinant PfLDH-based enzymatic activity determination provides a simplified, reproductive and economic platform for screening inhibitors of PfLDH, which contributes to the discovery of inhibitory compounds from either pure chemicals or crude extracts of medicinal plants, animals, and fungi towards development of novel antimalarial drugs.

Chuang Chen,1 Jing-Ping Yuan,2,3 Wen Wei,1 Yi Tu,1 Feng Yao,1 Xue-Qin Yang,4 Jin-Zhong Sun,1 Sheng-Rong Sun,1 Yan Li2 1Department of Breast and Thyroid Surgery, Wuhan University, Renmin Hospital, Wuhan, 2Department of Oncology, Zhongnan Hospital of Wuhan University and Hubei Key Laboratory of Tumor Biological Behaviors and Hubei Cancer Clinical Study Center, Wuhan, 3Department of Pathology, The Central Hospital of Wuhan, Wuhan, 4Medical School of Jingchu University of Technology, Jingmen, People’s Republic of China Background: Hormone receptors, including the estrogen receptor and progesterone receptor, human epidermal growth factor receptor 2 (HER2), and other biomarkers like Ki67, epidermal growth factor receptor (EGFR, also known as HER1), the androgen receptor, and p53, are key molecules in breast cancer. This study evaluated the relationship between HER2 and hormone receptors and explored the additional prognostic value of Ki67, EGFR, the androgen receptor, and p53. Methods: Quantitative determination of HER2 and EGFR was performed in 240 invasive breast cancer tissue microarray specimens using quantum dot (QD)-based nanotechnology. We identified two subtypes of HER2, ie, high total HER2 load (HTH2) and low total HER2 load (LTH2), and three subtypes of hormone receptor, ie, high hormone receptor (HHR), low hormone receptor (LHR), and no hormone receptor (NHR). Therefore, breast cancer patients could be divided into five subtypes according to HER2 and hormone receptor status. Ki67, p53, and the androgen receptor were determined by traditional immunohistochemistry techniques. The relationship between hormone receptors and HER2 was investigated and the additional value of Ki67, EGFR, the androgen receptor, and p53 for prediction of 5-year disease-free survival was assessed. Results: In all patients, quantitative determination showed a statistically significant (P0.05) between the five subtypes. EGFR and Ki67 had prognostic significance for 5-year disease-free survival in univariate analysis, but the androgen receptor and p53 did not. Multivariate analysis identified that EGFR expression had predictive significance for 5-year disease-free survival in hormone-receptor positive patients and in those with the lymph node-positive breast cancer subtype. Conclusion: Hormone receptor expression was indeed one of the molecular profiles in the subtypes identified by quantitative HER2 and vice versa. EGFR status may provide discriminative prognostic information in addition to HER2 and hormone receptor status, and should be integrated into routine practice to help formulate more specific prediction of the prognosis and appropriate individualized treatment. Keywords: quantum dots, breast cancer, molecular classification, prognosis, prediction

Ki-67 is a biomarker that reflects the cell proliferation state. Despite a clear understanding of the protein's structure and properties, its functional role remains elusive. We conducted the present study to assess the prognostic value of Ki-67 in breast cancer (BC).We enrolled 164 individuals in this study: 30 patients with benign tumors and 134 with invasive BC. Immunohistochemistry (IHC) was used to detect Ki-67 expression The prognostic value of Ki-67 for 5-year recurrence-free survival (RFS) could be analyzed in 134 BC patients.Ki-67 expression showed significant differences with the tumor grade, lymph node (LN) status, HER2 status and hormone receptor (HR) status (all P0.05). When Ki- 67 11% was used as cutoff to divide the 134 cases into two groups, with high and low expression, the patients in former had a significantly higher 5-year recurrence rate (37.1% vs 8.1%, P=0.001) and a worse RFS (log-rank test, P=0.0017) than those in low Ki-67 expression group. Ki-67 was an independent prognostic predictor of 5-year RFS in both univariate and multivariate analyses.Ki-67 can be used as a negative predictor of 5-year RFS of patients with invasive BC.

The prevalence of prostate cancer (PCa) is high and PCa is the most common cutaneous cancer in men worldwide. Despite extensive research efforts, very few biomarkers of PCa have been introduced to date in clinical practice. A meta-analysis was performed on the most recently reported CTC to assess its prognostic effect and to elucidate whether its detection in the peripheral blood of patients diagnosed with metastatic, castration-resistant prostate cancer (CPRC) and Hormone Refractory Prostate Cancer (HRPC) can be used as a prognostic factor for survival.We searched Science Direct, EMBASE, PubMed, and Cell Research databases for studies that assessed the prognostic relevance of the presence number of circulating tumor cells (CTC) detection in the peripheral blood (PB). A fixed effects model with relative risk (RR) and 95% confidence interval (95% CI) is used for analysis.A total of 4 studies, including 486 patients, were eligible for final analysis. Pooled analysis indicated the presence number of CTC per 7.5 ml peripheral blood is associated with a poor survival rate (RR=2.51, 95% CI 1.96-3.21).The unfavorable count (presence of 5 or more CTCs per 7.5 ml peripheral blood) was associated with poor overall survival in patients with PCa. CTC counts can be used as an accurate and independent predictor of survival rate in patients with PCa.

Although crosstalk between cytosolic and plastidic terpenoid pathways has been validated in many plant species, we report here for the first time a striking elevation of the nucleus-encoded artemisinin biosynthesis relevant DBR2 mRNA following the incubation of plants with fosmidomycin (FM). FM decreased singlet oxygen (1O2) scavengers such as β-carotene and α-tocopherol and subsequently invoked 1O2 burst. The treatment of plants with fluridone (FD) neither decreased α-tocopherol content nor triggered 1O2 emission. In conclusion, FM can up-regulate 1O2-sensitive nuclear genes responsible for artemisinin biogenesis by mitigating the accumulation of plastidic scavenging terpenoids, thereby eliciting 1O2 generation and initiating 1O2 retrograde signaling.

Xue-Qin Yang1,2, Chuang Chen1, Chun-Wei Peng1, Jin-Xuan Hou1, Shao-Ping Liu1, Chu-Bo Qi3, Yi-Ping Gong4, Xiao-Bo Zhu5, Dai-Wen Pang6, Yan Li1 1Department of Oncology, Zhongnan Hospital of Wuhan University, Hubei Key Laboratory on Tumor Biological Behaviors and Hubei Cancer Clinical Study Center, Wuhan; 2Medical School of Jingchu University of Technology, Jingmen; 3Department of Pathology; 4Department of Breast Surgery, Hubei Cancer Hospital, Wuhan; 5Wuhan Jiayuan Quantum Dots Co Ltd and Wuhan Tumor Nanometer Diagnosis Engineering Research Center, Wuhan; 6Key Laboratory of Analytical Chemistry for Biology and Medicine, College of Chemistry and Molecular Sciences, and State Key Laboratory of Virology, Wuhan University, Wuhan, People's Republic of China Background: The epidermal growth factor receptor (EGFR) is a promising therapeutic target in cancer, but its clinical value in breast cancer remains controversial. Our previous studies have found that quantitative analysis of biomarkers with quantum dot-based nanotechnology had better detection performance than conventional immunohistochemistry. The present study was undertaken to investigate the prognostic value of EGFR in breast cancer using quantum dot-based quantitative spectral analysis. Methods: EGFR expression in 65 breast cancer specimens was detected by immunohistochemistry and quantum dot-immunohistochemistry, and comparisons were made between the two methods. EGFR expression in tissue microarrays of 240 breast cancer patients was then detected by quantum dot-immunohistochemistry and spectral analysis. The prognostic value of EGFR immunofluorescence area (EGFR area) for five-year recurrence-free survival was investigated. Results: The same antigen localization, high correlation of staining rates (r = 0.914), and high agreement of measurement (κ= 0.848) of EGFR expression in breast cancer were found by quantum dot-immunohistochemistry and immunohistochemistry. The EGFR area showed significant differences by tumor grade, lymph node status, HER2 status, and hormone receptor status (all P < 0.05). Patients in the large EGFR area (≥30.51) group had a significantly higher five-year recurrence rate (47.2% versus 27.4%, P = 0.002) and worse five-year recurrence-free survival (log-rank test, P = 0.0015) than those in the small EGFR area (