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A wide array of microorganisms, including many novel, phylogenetically deeply rooted taxa, survive and thrive in extreme environments. These unique and reduced-complexity ecosystems offer a tremendous opportunity for studying the structure, function and evolution of natural microbial communities. Marker gene surveys have resolved patterns and ecological drivers of these extremophile assemblages, revealing a vast uncultured microbial diversity and the often predominance of archaea in the most extreme conditions. New omics studies have uncovered linkages between community function and environmental variables, and have enabled discovery and genomic characterization of major new lineages that substantially expand microbial diversity and change the structure of the tree of life. These efforts have significantly advanced our understanding of the diversity, ecology and evolution of microorganisms populating Earth’s extreme environments, and have facilitated the exploration of microbiota and processes in more complex ecosystems. Microbial life can thrive in extreme environments such as terrestrial hot springs and deep sea hydrothermal vents, glaciers and permafrost, hypersaline habitats, acid mine drainage and the subsurface. In this Review, Shu and Huang explore the diversity, functions and evolution of bacteria and archaea inhabiting Earth’s major extreme environments.

Ningning Yang,1,2 Hongchun Chen,1,2 Yuchen Huang,1,2 Xuexue Song,3 Panpan Yang,4 Shan Zhang,5 Wentian Yan,1,2 Nan Li,1,2 Zhenzhong Feng1,2 1Department of Pathology, First Affiliated Hospital of Bengbu Medical College, Bengbu, 233000, People’s Republic of China; 2Department of Pathology, Bengbu Medical College, Bengbu, 233000, People’s Republic of China; 3Department of Pathology, First Affiliated Hospital of University of Science and Technology of China, Hefei, 230000, People’s Republic of China; 4Department of Pathology, Second Affiliated Hospital of Anhui Medical University, Hefei, 230000, People’s Republic of China; 5Department of Pathology, Second People’s Hospital of Hefei, Hefei, 230000, People’s Republic of ChinaCorrespondence: Zhenzhong Feng; Nan LiDepartment of Pathology, First Affiliated Hospital of Bengbu Medical College, No. 287 Changhuai Road, Bengbu, Anhui Province, People’s Republic of ChinaTel +86 15055289508Email fzz18297301626@163.com; linanangel100@sina.comPurpose: As a non-classical ligand of Wnt, the abnormal regulation of Wnt5a contributes to the progression of malignant tumors; however, its effects differ depending on tumor type. Here, we evaluated the expression and significance of Wnt5a in endometrioid adenocarcinoma and its relationship with epithelial–mesenchymal transition (EMT)-related proteins.Patients and Methods: Immunohistochemical streptavidin-peroxidase method and reverse transcription polymerase chain reaction (RT-PCR) method were used to analyze the expression and correlation of Wnt5a, and EMT-related protein β-catenin, E-cadherin and enhancer of zeste homolog 2 (EZH2) in endometrial cancer tissues and cell samples of each group.Results: The expression of Wnt5a and E-cadherin decreased in the following order, normal endometrium > atypical hyperplasia endometrium > endometrioid adenocarcinoma. In contrast, the expression of β-catenin and EZH2 increased gradually. Moreover, Wnt5a expression was associated with the degree of tissue differentiation, International Federation of Gynecology and Obstetrics (FIGO) stage, and lymph node metastasis (all P< 0.05). Wnt5a expression was also negatively correlated with β-catenin and EZH2 expression and positively correlated with E-cadherin expression. RT-PCR results further indicated that E-cadherin mRNA expression was upregulated in a Wnt5a-overexpressing Ishikawa cell line compared to cells transfected with an empty vector or negative control cells (P< 0.01). Furthermore, the expression of EZH2 and β-catenin mRNA was downregulated in overexpressing cells compared to empty vector and negative control cells (P< 0.01).Conclusion: Wnt5a may elicit a suppressive effect on endometrioid adenocarcinoma by inhibiting EMT. This study provides a theoretical basis for the pathological diagnosis and targeted therapy of endometrioid adenocarcinoma and extends our understanding of the Wnt5a signaling pathway.Keywords: endometrioid adenocarcinoma, Wnt5a, epithelial–mesenchymal transition, immunohistochemistry

Candida tropicalis can metabolize alkanes or fatty acids to produce long-chain dicarboxylic acids (DCAs). Fatty acid transporters located on the cell or peroxisome membrane may play an important role in this process. Using amino acid sequence homologous alignment, two putative proteins, CtFat1p and CtPxa1p, located on the cell and peroxisome membrane were found, respectively. Moreover, single- and double-knockout homologous recombination technology was used to study ctfat1p and ctpxa1p gene effects on DCA synthesis. In comparison to the wild-type strain, long-chain DCA yield decreased by 65.14%, 88.38% and 56.19% after single and double-copy knockout of ctfat1p genes and double-copy knockout of ctpxa1p genes, respectively, indicating that the knockout of ctfat1p and ctpxa1p genes had a significant effect on the conversion of oils and fats into long-chain DCAs by C. tropicalis. However, the yield of long-chain DCAs increased by 21.90% after single-knockout of the ctpxa1p gene, indicating that the single-knockout of the ctpxa1p gene may reduce fatty acid transport to peroxisome for further oxidation. Moreover, to improve the intracellular transport rate of fatty acids, ctfat1p copy number increased, increasing DCA yield by 30.10%. These results may provide useful information for enhancing the production of long-chain DCAs by C. tropicalis.

Human respiratory syncytial virus (RSV) infection is the leading cause of lower respiratory tract illness (LRTI), and no vaccine against LRTI has proven to be safe and effective in infants. Our study assessed attenuated recombinant RSVs as vaccine candidates to prevent RSV infection in mice. The constructed recombinant plasmids harbored (5′ to 3′) a T7 promoter, hammerhead ribozyme, RSV Long strain antigenomic cDNA with cold-passaged (cp) mutations or cp combined with temperature-sensitive attenuated mutations from the A2 strain (A2cpts) or further combined with SH gene deletion (A2cptsΔSH), HDV ribozyme (δ), and a T7 terminator. These vectors were subsequently co-transfected with four helper plasmids encoding N, P, L, and M2-1 viral proteins into BHK/T7-9 cells, and the recovered viruses were then passaged in Vero cells. The rescued recombinant RSVs (rRSVs) were named rRSV-Long/A2cp, rRSV-Long/A2cpts, and rRSV-Long/A2cptsΔSH, respectively, and stably passaged in vitro, without reversion to wild type (wt) at sites containing introduced mutations or deletion. Although rRSV-Long/A2cpts and rRSV-Long/A2cptsΔSH displayed temperature-sensitive (ts) phenotype in vitro and in vivo, all rRSVs were significantly attenuated in vivo. Furthermore, BALB/c mice immunized with rRSVs produced Th1-biased immune response, resisted wtRSV infection, and were free from enhanced respiratory disease. We showed that the combination of ΔSH with attenuation (att) mutations of cpts contributed to improving att phenotype, efficacy, and gene stability of rRSV. By successfully introducing att mutations and SH gene deletion into the RSV Long parent and producing three rRSV strains, we have laid an important foundation for the development of RSV live attenuated vaccines.

Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are the most typical pathogenic bacteria with a significantly high risk of bio-contamination, widely existing in hospital and public places. Recent studies on antibacterial materials and the related mechanisms have attracted more interests of researchers. However, the antibacterial behavior of materials is usually evaluated separately on the single bacterial strain, which is far from the practical condition. Actually, the interaction between the polymicrobial communities can promote the growing profile of bacteria, which may weaken the antibacterial effect of materials. In this work, a 420 copper-bearing martensitic stainless steel (420CuSS) was studied with respect to its antibacterial activity and the underlying mechanism in a co-culturing infection model using both E. coli and S. aureus. Observed via plating and counting colony forming units (CFU), Cu releasing, and material characterization, 420CuSS was proved to present excellent antibacterial performance against the mixed bacteria with an approximately 99.4 % of antibacterial rate. In addition, 420CuSS could effectively inhibit the biofilm formation on its surfaces, resulting from a synergistic antibacterial effect of Cu ions, Fe ions, reactive oxygen species (ROS), and proton consumption of bacteria.

Adipose-derived stem cell (ADSC)-based therapeutic strategies are in fast-pace advancement in wound treatment due to their availability and the ability to self-renew, undergo multilineage differentiation and self-renewal. Existing studies have successfully explored ADSCs to facilitate scar-free healing of small wounds, but whether the healing of large-area wounds that exhibit over 50% of skin tissue loss in the entire body could be achieved remains controversial. This study sought to review the mechanism of physiological wound healing, and discuss the roles played by chemokines, biological factors and biomaterial scaffolds. The possibility of applying ADSC-conditioned medium or ADSC-released exosomes as ‘off-the-shelf’ tissue engineering products, integrated with biomaterial scaffolds to facilitate wound healing, was analyzed.

Metabotropic GABAB G protein-coupled receptor functions as a mandatory heterodimer of GB1 and GB2 subunits and mediates inhibitory neurotransmission in the central nervous system. Each subunit is composed of the extracellular Venus flytrap (VFT) domain and transmembrane (TM) domain. Here we present cryo-EM structures of full-length human heterodimeric GABAB receptor in the antagonist-bound inactive state and in the active state complexed with an agonist and a positive allosteric modulator in the presence of Gi1 protein at a resolution range of 2.8–3.0 A. Our structures reveal that agonist binding stabilizes the closure of GB1 VFT, which in turn triggers a rearrangement of TM interfaces between the two subunits from TM3-TM5/TM3-TM5 in the inactive state to TM6/TM6 in the active state and finally induces the opening of intracellular loop 3 and synergistic shifting of TM3, 4 and 5 helices in GB2 TM domain to accommodate the α5-helix of Gi1. We also observed that the positive allosteric modulator anchors at the dimeric interface of TM domains. These results provide a structural framework for understanding class C GPCR activation and a rational template for allosteric modulator design targeting the dimeric interface of GABAB receptor.

Background As a rare subtype of prostate carcinoma, basal cell carcinoma (BCC) has not been studied extensively and thus lacks systematic molecular characterization. Methods Here, we applied single-cell genomic amplification and RNA-Seq to a specimen of human prostate BCC (CK34βE12+ /P63+ /PAP- /PSA- ). The mutational landscape was obtained via whole exome sequencing of the amplification mixture of 49 single cells, and the transcriptomes of 69 single cells were also obtained. Results The five putative driver genes mutated in BCC are CASC5, NUTM1, PTPRC, KMT2C, and TBX3, and the top three nucleotide substitutions are C>T, T>C, and C>A, similar to common prostate cancer. The distribution of the variant allele frequency values indicated that these single cells are from the same tumor clone. The 69 single cells were clustered into tumor, stromal, and immune cells based on their global transcriptomic profiles. The tumor cells specifically express basal cell markers like KRT5, KRT14, and KRT23 and epithelial markers EPCAM, CDH1, and CD24. The transcription factor covariance network analysis showed that the BCC tumor cells have distinct regulatory networks. By comparison with current prostate cancer datasets, we found that some of the bulk samples exhibit basal cell signatures. Interestingly, at single-cell resolution the gene expression patterns of prostate BCC tumor cells show uniqueness compared with that of common prostate cancer-derived circulating tumor cells. Conclusions This study, for the first time, discloses the comprehensive mutational and transcriptomic landscapes of prostate BCC, which lays a foundation for the understanding of its tumorigenesis mechanism and provides new insights into prostate cancers in general.

Gastroesophageal reflux disease (GERD) is a disease that stomach contents continually refluxing into esophagus causes symptoms and/or complications. The study was working to find natural plant extracts with good effects and small side effects to treat reflux esophagitis (RE). The anti-inflammatory effects of hexane extract of Magnolia sieboldii (MsHE) were conducted on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The ameliorative effects of MsHE on esophageal damage in rats induced by gastric acid reflux was explored in vivo. The results showed that MsHE decreased the production of nitric oxide (NO) and expression levels of iNOS, COX-2 and TNF-α on LPS-stimulated RAW 264.7 cells and MsHE treatment ameliorated the rats' esophageal tissue damage induced by gastric acid and inhibited the increase of inflammatory mediators and pro-inflammatory cytokines by regulating NF-κB signaling pathway. In addition, MsHE protected the function of barrier of epithelial cells against inflammatory conditions through increasing the expression of tight junctions. Furthermore, liquid chromatography-mass spectrometry analysis was used for determine the active ingredients contained in MsHE. The results show that MsHE can alleviate experimental rat RE by regulating NF-κB signaling pathway. In summary, MsHE may be used as a source material of drug candidate for the treatment of RE.

Butanol is an attractive second-generation biofuel as well as an important chemical. Production of butanol from lignocellulosic feedstock via consolidated bioprocessing has attracted increasing int...

Sea buckthorn (SB), also named sea berry, Hippophae rhamnoides L. or Elaeagnus rhamnoides L., has been used in daily life for centuries with kinds of purposes ranging from a beverage with a pleasant taste and flavor, to an agent for treatment of many disorders and diseases. SB is well known more than just a fruit. So far, a unique mixture of bioactive components was elucidated in SB including flavonoids, phenolic acids, proanthocyanidins, carotenoids, fatty acids, triterpenoids, vitamins and phytosterols, which implied the great medicinal worth of this seaberry. Both in vitro and in vivo experiments, ranged from cell lines to animals as well as a few in patients and healthy volunteers, indicated that SB possessed various biological activities including anti-inflammatory and immunomodulatory effects, antioxidant properties, anti-cancer activities, hepato-protection, cardiovascular-protection, neuroprotection, radioprotection, skin protection effect as well as the protective effect against some eye and gastrointestinal sickness. Furthermore, the toxicological results revealed neither the fruits, nor the seeds of SB were toxic. The present review summarizes the unique profile of the chemical compounds, the nutritional and health effects as well as the toxicological properties of SB, which lay the foundation for practical applications of SB in treatment of human diseases.

The European physical journal / C 81(11), 973 (2021). doi:10.1140/epjc/s10052-021-09544-4, The OSIRIS detector is a subsystem of the liquid scintillator filling chain of the JUNO reactor neutrino experiment. Its purpose is to validate the radiopurity of the scintillator to assure that all components of the JUNO scintillator system work to specifications and only neutrino-grade scintillator is filled into the JUNO Central Detector. The aspired sensitivity level of $10^{���16}$ g/g of $^{238}$U and $^{232}$Th requires a large (���20 m$^3$) detection volume and ultralow background levels. The present paper reports on the design and major components of the OSIRIS detector, the detector simulation as well as the measuring strategies foreseen and the sensitivity levels to U/Th that can be reached in this setup., Published by Springer, Heidelberg

Enzymatic production of xylitol is a promising alternative to the chemical hydrogenation process. However, it encounters problems that are largely due to protein susceptibility to environmental factors. In this study, to develop a robust, practical enzymatic process for xylitol production, a coupled enzyme system consisting of formate dehydrogenase (FDH), glucose dehydrogenase (GDH), and xylose reductase (XR) was constructed, wherein the alkaline product produced by FDH and the acidic product produced by GDH could neutralize each other during cofactor regeneration. After optimization of conditions, a pH-neutralization, redox-balanced process was developed that could be carried out in pure water requiring no pH regulation. As a result, a xylitol production of 273.6 g/L that is much higher than those yet reported was obtained from 2 M xylose in 24 h, with a relatively high productivity of 11.4 g/(L h). The strategy demonstrated here can be adapted for the production of other NADH-consuming products.

Background and aims Aristolochic acid (AA) exposure has been statistically associated with human liver cancers. However, direct evidence of AA exposure-induced liver cancer is absent. This study aims to establish a direct causal relationship between AA exposure and liver cancers based on a mouse model and then explores the AA-mediated genomic alterations that could be implicated in human cancers with AA-associated mutational signature. Approach and results We subjected mice, including phosphatase and tensin homolog (Pten)-deficient ones, to aristolochic acid I (AAI) alone or a combination of AAI and CCl4 . Significantly, AAI exposure induced mouse liver cancers, including hepatocellular carcinoma (HCC) and combined HCC and intrahepatic cholangiocarcinoma, in a dose-dependent manner. Moreover, AAI exposure also enhanced tumorigenesis in these CCl4 -treated or Pten-deficient mice. AAI led to DNA damage and AAI-DNA adduct that could initiate liver cancers through characteristic adenine-to-thymine transversions, as indicated by comprehensive genomic analysis, which revealed recurrent mutations in Harvey rat sarcoma virus oncogene. Interestingly, an AA-associated mutational signature was mainly implicated in human liver cancers, especially from China. Moreover, we detected the AAI-DNA adduct in 25.8% (16/62) of paratumor liver tissues from randomly selected Chinese patients with HCC. Furthermore, based on phylogenetic analysis, the characteristic mutations were found in the initiating malignant clones in the AA-implicated mouse and human liver cancers where the mutations of tumor protein p53 and Janus kinase 1 were prone to be significantly enriched in the AA-affected human tumors. Conclusions This study provides evidence for AA-induced liver cancer with the featured mutational processes during malignant clonal evolution, laying a solid foundation for the prevention and diagnosis of AA-associated human cancers, especially liver cancers.

Background Rehmannia glutinosa polysaccharide is the main reason that contributes to the immunological function of R. glutinosa. Due to its disadvantages in clinical use, here we designed the PEGylation nano-RGP (pRL) to improve the drug-targeting effect and the immunological function. Our present work aims to establish the optimum condition of preparing the pRL and to investigate its immunological function on macrophages. Methods pRL was prepared by thin film hydration method combined with ultra-sonication technique. And its preparation conditions were optimized with response surface methodology. Also, the lyophilization method was optimized. The characteristics of the pRL were evaluated, including particle size, drug loading, encapsulation efficiency and morphology. The immunological function of pRL on macrophage was investigated through CCK-8 test, ELISA and flow cytometry. Results The lipid-to-cholesterol molar ratio of 8:1, the addition of DSPE-PEG2000 of 9% and the lipid-to-drug ratio of 5.4:1 were the optimum preparation technology for pRL. The encapsulation efficiency (EE) of pRL under this preparation technology was 95.81±1.58%, with a particle size of 31.98 ± 2.6 nm. The lactose-to-lipid ratio (2:1) was the optimal lyophilization method. pRL promoted macrophage proliferation, which is significantly better than that of nano-RGP without PEGylation (RL). pRL-stimulated RAW264.7 cells showed a high secretion of pro-inflammatory cytokines, which is the characteristic indicator of M1 polarization. Enhanced cellular uptake through macropinocytosis-dependent and caveolae-mediated endocytosis was observed in pRL-treated RAW264.7 cells. Conclusion Our study concluded that PEGylation effectively overcame the poor targeting effect of Rehmannia glutinosa polysaccharide (RGP) and significantly improved the immunological profile of its nano-formulation, which suggested that pRL could serve as an immune adjuvant in clinical application.

Glioblastoma multiforme (GBM) is the most common and malignant brain tumor, and almost half of the patients carrying EGFR-driven tumor with PTEN deficiency are resistant to EGFR-targeted therapy. EGFR amplification and/or mutation is reported in various epithelial tumors. This series of studies aimed to identify a potent compound against EGFR-driven tumor. We screened a chemical library containing over 600 individual compounds purified from Traditional Chinese Medicine against GBM cells with EGFR amplification and found that cinobufagin, the major active ingredient of Chansu, inhibited the proliferation of EGFR amplified GBM cells and PTEN deficiency enhanced its anti-proliferation effects. Cinobufagin also strongly inhibited the proliferation of carcinoma cell lines with wild-type or mutant EGFR expression. In contrast, the compound only weakly inhibited the proliferation of cancer cells with low or without EGFR expression. Cinobufagin blocked EGFR phosphorylation and its downstream signaling, which additionally induced apoptosis and cytotoxicity in EGFR amplified cancer cells. In vivo, cinobufagin blocked EGFR signaling, inhibited cell proliferation, and elicited apoptosis, thereby suppressing tumor growth in both subcutaneous and intracranial U87MG-EGFR xenograft mouse models and increasing the median survival of nude mice bearing intracranial U87MG-EGFR tumors. Cinobufagin is a potential therapeutic agent for treating malignant glioma and other human cancers expressing EGFR.

Recent omics studies have provided invaluable insights into the metabolic potential, adaptation, and evolution of novel archaeal lineages from a variety of extreme environments. We utilized a genome-resolved metagenomic approach to recover eight medium- to high-quality metagenome-assembled genomes (MAGs) that likely represent a new order ("Candidatus Sysuiplasmatales") in the class Thermoplasmata from mine tailings and acid mine drainage (AMD) sediments sampled from two copper mines in South China. 16S rRNA gene-based analyses revealed a narrow habitat range for these uncultured archaea limited to AMD and hot spring-related environments. Metabolic reconstruction indicated a facultatively anaerobic heterotrophic lifestyle. This may allow the archaea to adapt to oxygen fluctuations and is thus in marked contrast to the majority of lineages in the domain Archaea, which typically show obligately anaerobic metabolisms. Notably, "Ca. Sysuiplasmatales" could conserve energy through degradation of fatty acids, amino acid metabolism, and oxidation of reduced inorganic sulfur compounds (RISCs), suggesting that they may contribute to acid generation in the extreme mine environments. Unlike the closely related orders Methanomassiliicoccales and "Candidatus Gimiplasmatales," "Ca. Sysuiplasmatales" lacks the capacity to perform methanogenesis and carbon fixation. Ancestral state reconstruction indicated that "Ca. Sysuiplasmatales," the closely related orders Methanomassiliicoccales and "Ca. Gimiplasmatales," and the orders SG8-5 and RBG-16-68-12 originated from a facultatively anaerobic ancestor capable of carbon fixation via the bacterial-type H4F Wood-Ljungdahl pathway (WLP). Their metabolic divergence might be attributed to different evolutionary paths. IMPORTANCE A wide array of archaea populate Earth's extreme environments; therefore, they may play important roles in mediating biogeochemical processes such as iron and sulfur cycling. However, our knowledge of archaeal biology and evolution is still limited, since the majority of the archaeal diversity is uncultured. For instance, most order-level lineages except Thermoplasmatales, Aciduliprofundales, and Methanomassiliicoccales within Thermoplasmata do not have cultured representatives. Here, we report the discovery and genomic characterization of a novel order, "Ca. Sysuiplasmatales," within Thermoplasmata in extremely acidic mine environments. "Ca. Sysuiplasmatales" are inferred to be facultatively anaerobic heterotrophs and likely contribute to acid generation through the oxidation of RISCs. The physiological divergence between "Ca. Sysuiplasmatales" and closely related Thermoplasmata lineages may be attributed to different evolutionary paths. These results expand our knowledge of archaea in the extreme mine ecosystem.

Cadmium (Cd) and arsenic (As) contamination in paddy soils poses serious health risks to humans. The accumulation of Cd and As in rice (Oryza sativa L.) depends on their bioavailability, which is affected by soil physicochemical properties and soil microbial activities. However, little is known about the intricate interplay between rice plants and their rhizosphere microbes during the uptake of Cd and As. In this study, different bacterial communities were established by sterilizing paddy soils with γ-radiation. A pot experiment using two paddy soils with different levels of contamination was conducted to explore how the bacterial community composition affects Cd and As accumulation in rice plants. The results showed that the sterilization treatment substantially changed the bacterial composition in the rhizosphere, and significantly increased the grain yield (by 33.5–38.3%). The sterilization treatment resulted in significantly decreased concentrations of Cd (by 18.2–38.7%) and As (by 20.3–36.7%) in the grain, straw, and root of rice plants. The accumulation of Cd and As in rice plants was negatively correlated with the relative abundance of sulfate-reducing bacteria and iron-oxidizing bacteria in the rhizosphere. Other specific taxa associated with the accumulation of Cd and As in rice plants were also identified. Our results suggest that regulating the composition of the rhizosphere bacterial community could simultaneously reduce Cd and As accumulation in rice grain and increase the grain yield. These results would be useful for developing strategies to cultivate safe rice crops in areas contaminated with Cd and As.

Melanocortin-4 receptor (MC4R) plays a central role in the regulation of energy homeostasis. Its high sequence similarity to other MC receptor family members, low agonist selectivity and the lack of structural information concerning MC4R-specific activation have hampered the development of MC4R-seletive therapeutics to treat obesity. Here, we report four high-resolution structures of full-length MC4R in complex with the heterotrimeric G(s) protein stimulated by the endogenous peptide ligand α-MSH, FDA-approved drugs afamelanotide (Scenesse™) and bremelanotide (Vyleesi™), and a selective small-molecule ligand THIQ, respectively. Together with pharmacological studies, our results reveal the conserved binding mode of peptidic agonists, the distinctive molecular details of small-molecule agonist recognition underlying receptor subtype selectivity, and a distinct activation mechanism for MC4R, thereby offering new insights into G protein coupling. Our work may facilitate the discovery of selective therapeutic agents targeting MC4R.

The experiments were conducted at Key Laboratory of Genetics and Physiology of Yangzhou University. The conventional cultivar Sikang-1 and hybrid cultivar Sikang-3 were used as the experimental materials. The potted cotton at squaring stage was moved to artificial climatic chamber with 38°C for 72 h , and 38°C for 96 h respectively in 2017 and 2018 cotton growth season, and moved to natural conditions, then the squares were collected to measure the Bt protein concentration and nitrogen metabolic physiology at 0, 12, 24, 48, 72, 96 hours respectively after the high temperature termination. The square Bt insecticidal protein contents were able to recover to the corresponding control level, the time for the recovery was in need of longer as the stressed high temperature period increased. In 72 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 72 h for Sikang-1 and at 48 h for Sikang-3. In 96 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 96 h for Sikang-1 and at 72 h for Sikang-3. There were significantly positive correlations of Bt toxin content with soluble protein content, glutamate pyruvate transaminase activity and glutamate oxaloacetate transaminase activity (the correlation coefficients were 0.964**, 0.981**, and 0.971**, respectively), and significantly negative correlations of Bt toxin content with free amino acid contents, and activitied of protease and peptidase (the correlation coefficients were -0.894**, -0.912**, and -0.834**, respectively). Therefore, the recovery degree of Bt insecticidal protein content of cotton square can be predicted according to the duration of high temperature stress, reasonably controlling Helicoverpa armigera and related pests in production.

We evaluated the influence of different proportions of 5′‐inosine monophosphate (IMP) and 5′‐guanosine monophosphate (GMP) on growth, feed digestibility and activity of digestive enzymes of turbot Scophthalmus maximus. Weight gain and daily feed intake were significantly higher in S. maximus fed with IMP or GMP, in comparison with fish fed with neither IMP nor GMP. The growth of 0.05% IMP + 0.05% GMP group was the best, and the intestinal digestive function was improved. The addition of IMP and GMP to fish diets significantly increased the apparent feed digestibility coefficient of dry matter and protein, as well as intestinal protease activity. The highest intestinal protease activity was observed in fish fed with 1 g/kg IMP. However, the lipase activity in hepatopancreas decreased significantly after addition of nucleic acid. According to our results, the optimal level of dietary IMP is 1 g/kg, which is in line with most of the growth performance and feed digestibility.

Strain SYSU-17, representing a novel acid-tolerant yeast species which can grow at pH 2.0 weakly, was isolated from acid mine drainage collected in a tailing impoundment of the Fankou Lead/Zinc Mine, Guangdong Province, PR China. Phylogenetic analysis of strain SYSU-17 based on the internal transcribed spacer (ITS) region and the D1/D2 domains of the large subunit ribosomal RNA (LSU rRNA) gene suggested that strain SYSU-17 was a novel species belonging to the genus Spencerozyma (class Microbotryomycetes, subphylum Pucciniomycotina). It differed from the type strain of the closest related species, Spencerozyma crocea CBS 2029T, by 0.7 % sequence divergence (three gaps and one nucleotide substitution out of 594 bp) in the D1/D2 domains of the LSU rRNA gene and 7.6 % sequence divergence (32 gaps and 22 nucleotide substitutions out of 714 bp) in the ITS region. In contrast to the physiological properties of S. crocea, the novel yeast species was unable to assimilate galactose, d-ribose, xylitol, succinate, d-xylose, ethanol, nitrate and nitrite. The name Spencerozyma acididurans sp. nov. is proposed and SYSU-17 is designated as the holotype.

Liaodong Bay is one of the largest semi-enclosed bays located in Bohai Sea, in northeast China. It is an important feeding and spawning place, even wintering and breeding migration base for many marine organisms. The size structure of phytoplankton communities is a key feature of marine ecosystems and can be used as an indicator for algae disaster, but the methods to evalute is still challenging. Here, we compared the size-fractionated chlorophyll-a method with a high-throughout sequencing method in an evaluation of the size structure of phytoplankton assemblages in Liaodong Bay, China, in 2014. The proportion of picophytoplankton was estimated at just 19% according to the results based on size-fractionated chlorophyll-a, which likely represents a severe underestimation. The sequencing method confirmed that many kinds of phytoplankton species appeared in more than one size-fractionated filter membrane, with some even dominating where the filter membrane pore size was greater than the individual phytoplankton size. According to the sequencing method, the phytoplankton assemblages in Liaodong Bay were dominated by picophytoplankton (average proportion = 44%), followed by the nanophytoplankton (average 32%), a composition that is more consistent with other studies. The sequencing method provides a more robust way to assess the size structure and taxonomic diversity of marine phytoplankton communities.

Drought is a major limiting factor to maize (Zea mays L.) yield. Plant hormones, including gibberellins (GAs), play important roles in plant response to drought stress. In previous studies, significant reductions in GAs levels have been reported under drought stress. In maize, GA content is correlated to drought tolerance, but the molecular mechanism remains unclear. In the present study, AtG2ox1, a member of the GA2ox family with a clear function, was used to create GA deficiency maize. The transgenic maize had a higher chlorophyll content and faster growth rate, when compared to the wild type (WT) plants, under drought stress in a greenhouse. The physiological and biochemical test results revealed that transgenic maize had decreased levels of GA1 and malondialdehyde (MDA), and increased content of proline and soluble sugars, and antioxidant enzyme activities, when compared to the WT. Furthermore, the transcriptomic analysis revealed that some differentially expressed genes involved in transcription factors correlated to drought stress and abiotic stress responses, and that signaling was enriched. All these results reveal the possible molecular mechanism of GA regulation in drought tolerance, in which the overexpression of AtGA2ox1 altered the expression of multiple genes correlated to the internal antioxidant system and maintenance of cell osmotic potential. The present study demonstrates that the overexpression of AtGA2ox1 could control GA content and improve drought tolerance in transgenic maize. Furthermore, this strategy represents a novel approach to address drought tolerance in maize breeding.

Brown tides have occurred in the Bohai Sea of China since 2009 and caused serious losses in aquaculture. In recent years, phytoplankton identification has been greatly promoted by the development of high-throughput sequencing technologies. In this study, V4 region of 18S rDNA was used as the target gene to design the primer V4 (F/R), and high-throughput sequencing technology was adopted to evaluate the diversity of composition and characteristics of the eukaryotic phytoplankton community. A total of 93 alga species were identified, and about 60% (56 out of 93) of them were undetected or unreported in Chinese waters. There were 59% of the total algal species identified as autotrophy in trophic mode while heterotrophy and mixotrophy accounted for 12% and 29% of the total algal species, respectively. 8 species of picophytoplankton that were mainly distributed in the south and a partial area of the east and west formed the scientific basis for the quality and safety early-warning in aquatic products and brown rides study. Diversity index analysis also revealed a trend of fluctuations of marine phytoplankton diversity in different sample sites in Liaodong Bay. In conclusion, our work provides a detailed evaluation of eukaryotic phytoplankton diversity in Liaodong Bay, suggesting the need for long-term monitoring to further study and guard against the potential risk of harmful algal blooms.

Several abundant but yet uncultivated bacterial groups exist in extreme iron- and sulfur-rich environments, and the physiology, biodiversity, and ecological roles of these bacteria remain a mystery. Here we retrieved four metagenome-assembled genomes (MAGs) from an artificial acid mine drainage (AMD) system, and propose they belong to a new deltaproteobacterial order, Candidatus Acidulodesulfobacterales. The distribution pattern of Ca. Acidulodesulfobacterales in AMDs across Southeast China correlated strongly with ferrous iron. Reconstructed metabolic pathways and gene expression profiles showed that they were likely facultatively anaerobic autotrophs capable of nitrogen fixation. In addition to dissimilatory sulfate reduction, encoded by dsrAB, dsrD, dsrL, and dsrEFH genes, these microorganisms might also oxidize sulfide, depending on oxygen concentration and/or oxidation reduction potential. Several genes with homology to those involved in iron metabolism were also identified, suggesting their potential role in iron cycling. In addition, the expression of abundant resistance genes revealed the mechanisms of adaptation and response to the extreme environmental stresses endured by these organisms in the AMD environment. These findings shed light on the distribution, diversity, and potential ecological role of the new order Ca. Acidulodesulfobacterales in nature.

The purpose of this study was to develop a rapid method based on a real-time PCR assay designed to identify the presence of pufferfish in roasted fish fillet. Specific primers and probes were designed targeting Takifugu spp. and Lagocephalus spp., the most common genera in China. Specificity and sensitivity of this assay design were tested by using artificially spikes of pufferfish mixed in with other fish, such as Gadus and Thamnaconus septentrionalis,among others. Fifteen samples of retail roasted fish fillet and six samples from a 1999 poisoning event that occurred in Fujian province China were analysed for pufferfish. When the assay design was validated, no cross-reaction was observed between pufferfish and other species of fish. The limit of detection (LOD) was determined to be 0.001 ng pufferfish template, and the sensitivity of the method was 1%. Lagocephalus lunari was detected in six samples assayed from 1999 and no pufferfish was detected in the 15 retail roasted fish fillet samples tested. These results showed that the method was efficient for screening for pufferfish contamination in the roasted fish fillet and it could benefit public health protection by reducing the risk of tetrodotoxin poisoning.

Genetic heterogeneity of tumor is closely related to its clonal evolution, phenotypic diversity and treatment resistance, and such heterogeneity has only been characterized at single-cell sub-chromosomal scale in liver cancer. Here we reconstructed the single-variant resolution clonal evolution in human liver cancer based on single-cell mutational profiles. The results indicated that key genetic events occurred early during tumorigenesis, and an early metastasis followed by independent evolution was observed in primary liver tumor and intrahepatic metastatic portal vein tumor thrombus. By parallel single-cell RNA-Seq, the transcriptomic phenotype of HCC was found to be related with genetic heterogeneity. For the first time we reconstructed the single-cell and single-variant clonal evolution in human liver cancer, and dissection of both genetic and phenotypic heterogeneity will facilitate better understanding of their relationship.

Background Excessive and continuous inflammation may be the main cause of various immune system diseases. Reflux esophagitis (RE) is a common gastroesophageal reflux disease (GERD). Camellia japonica has high medicinal value and has long been used as a traditional herbal hemostatic medicine in China and Korea. The purpose of this study is to explore the antioxidant and anti-inflammatory activities of CJE and its protective effect on RE. Materials and methods Buds from C. japonica plants were collected in the mountain area of Jeju, South Korea. Dried C. japonica buds were extracted with 75% ethanol. DPPH and ABTS radical scavenging assay were evaluated according to previous method. The ROS production and anti-inflammatory effects of C. japonica buds ethanol extract (CJE) were evaluated on LPS-induced RAW 264.7 cell inflammation. The protective effects of CJE on RE were conducted in a RE rat model. Results CJE eliminated over 50% of DPPH and ABTS radical at concentration of 100 and 200 µg/mL, respectively. CJE alleviated changes in cell morphology, reduced production of ROS, NO and IL-1β. Also, down-regulated expression levels of iNOS, TNF-α, phosphorylated NF-κB, IκBα, and JNK/p38/MAPK. CJE reduced esophageal tissue damage ratio (40.3%) and attenuation of histological changes. In addition, CJE down-regulated the expression levels of TNF-α, IL-1β, COX-2 and phosphorylation levels of NF-κB and IκBα in esophageal tissue. Conclusions CJE possesses good anti-oxidation and anti-inflammatory activity, and can improve RE in rats caused by gastric acid reflux. Therefore, CJE is a natural material with good anti-oxidant and anti-inflammatory activity and has the possibility of being a candidate phytomedicine source for the treatment of RE.

Genetic heterogeneity of tumor is closely related to clonal evolution, phenotypic diversity and treatment resistance. Such heterogeneity has been characterized in liver cancer at single-cell sub-chromosomal scale, and a more precise single-variant resolution analysis is lacking. Here we employed a strategy to analyze both the single-cell genomic mutations and transcriptomic changes in 5 patients with liver cancer. Target sequencing was done for a total of 480 single cells in a patient-specific manner. DNA copy number status of point mutations was obtained from single-cell mutational profiling. The clonal structures of liver cancers were then uncovered at single-variant resolution, and mutation combinations in single cells enabled reconstruction of their evolutionary history. A common origin but independent evolutionary fate was revealed for primary liver tumor and intrahepatic metastatic portal vein tumor thrombus. The mutational signature suggested early evolutionary process may be related to specific etiology like aristolochic acids. By parallel single-cell RNA-Seq, the transcriptomic phenotype was found to be related with genetic heterogeneity in liver cancer. We reconstructed the single-cell and single-variant resolution clonal evolutionary history of liver cancer, and dissection of both genetic and phenotypic heterogeneity provides knowledge for mechanistic understanding of liver cancer initiation and progression.

In view of the effect of Lanthanum (La) on synaptic plasticity in offspring rats and its mechanism, 32 pregnant adult Wistar rats were divided into four groups randomly: control group (NC), 0.25%, 0.5% and 1.0% LaCl3 groups (n=8). All rats were poisoned through free drinking from day 0 of pregnancy to postnatal pregnancy to postnatal day 21 (suckling period). Morris was used to observe the spatial learning and memory ability of the offspring. The ultrastructure of hippocampus was observed by electron microscope. Expressions of synapsin, SYN, PSD95, NMDAR1, SHANK1, SHANK3 and GluR2 in the hippocampus were tested by qPCR and Western blot. The distribution and expression of functional synapses NMDAR1-SYN and PSD95 in CA1, CA3 and DG regions of hippocampus were analyzed by immunofluorescence. With increasing LaCl3 dose, the learning and memory abilities of offspring rats were significantly declined than those of NC group. Compared with NC group, there were abnormal changes in the ultrastructure of hippocampal tissue in LaCl3 groups, such as the shortening of synaptic active band, the thinning of postsynaptic density, and the decreasing of curvature of synaptic interface. The most obvious abnormal changes were found in the 1.0% LaCl3 group. The expressions of both mRNA and proteins (synapsin, SYN, PSD-95, NMDAR, SHANK1, SHANK3 and GluR2) in the hippocampus of the LaCl3 groups were significantly lower than those of NC group, and decreased with increasing LaCl3 dose. Compared with the control group, the co-expression of NMDAR1-SYN and the expression of PSD95 in CA1, CA3 and DG of hippocampus decreased significantly with the increase of Lanthanum exposure dose. Collectively, the mechanism of the decrease of learning and memory ability in offspring rats may be closely related to the abnormal expression of synaptic plasticity related proteins and the effect of Lanthanum exposure on synaptic transmission.

Recent genome-resolved metagenomic analyses of microbial communities from diverse environments have led to the discovery of many novel lineages that significantly expand the phylogenetic breadth of Archaea. Here, we report the genomic characterization of a new archaeal family based on five metagenome-assembled genomes retrieved from acid mine drainage sediments. Phylogenomic analyses placed these uncultivated archaea at the root of the candidate phylum Parvarchaeota, which expand this lesser-known phylum into two family levels. Genes involved in environmental adaptation and carbohydrate and protein utilization were identified in the ultra-small genomes (estimated size 0.53–0.76 Mb), indicating a survival strategy in this harsh environment (low pH and high heavy metal content). The detection of genes with homology to sulfocyanin suggested a potential involvement in iron cycling. Nevertheless, the absence of the ability to synthesize amino acids and nucleotides implies that these archaea may acquire these biomolecules from the environment or other community members. Applying evolutionary history analysis to Parvarchaeota suggested that members of the two families could broaden their niches by acquiring the potentials of utilizing different substrates. This study expands our knowledge of the diversity, metabolic capacity, and evolutionary history of the Parvarchaeota.

Heavy metal resistance is more than the tolerance one has towards a particular music genera [...]

This study examined the effects of the AKT/mTOR/HIF-1α signaling pathway on learning and memory in offspring rats induced by lanthanum from neuroethology and molecular biology perspectives. 32 pregnant adult Wistar rats were divided into four groups randomly: control group (NC), 0.25%, 0.5% and 1.0% LaCl3 groups (n = 8). All rats were poisoned through free drinking from day 0 of pregnancy to postnatal day 21 (suckling period). All offspring rats were poisoned through free drinking from delactation to postnatal day 48. Offspring rats aged 49-days-old were used as sampling objects to construct an LaCl3 poisoning model of offspring rats. Changes in hippocampal neurons, apoptosis of hippocampal neurons, learning and memory abilities of LaCl3-poisoned animals were measured by Nissl staining, TUNEL method and the shuttle box test, respectively. Expressions of PI3K, AKT, and mTOR, HIF-1α, and VEGF in the hippocampus were tested by qPCR and Western blot. Distributions of PI3K and p-AKT in hippocampal neurons were observed through the immunohistochemical method. With increasing LaCl3 dose, lightning strike time and active avoidance incubation period of offspring rats in the different LaCl3 groups were significantly prolonged. The Nissl body positive neurons of hippocampal neurons gradually declined while apoptosis in cells increased. The expressions of both mRNA (PI3K, AKT, mTOR, HIF-1α, VEGF) and proteins (PI3K, p-AKT, p-mTOR, HIF-1α, VEGF) in the hippocampus of the LaCl3 groups were significantly lower than those of NC group (p

Vaccines, in many cases, stimulate only too weak immunogenicity to prevent infection. Therefore, adjuvants are required during their preparation to boost the immune response. We herein developed a PEGylated nano-adjuvant based on Rehmannia glutinosa polysaccharide (RGP). The addition of PEG layer exhibits enhanced immune performance of the nano-RGP. Stimulation of dendritic cells (DCs) with PEGylated nano-RGP (pRL) led to increased proliferation and cytokine production (IL-6, IL-12, IL-1β and TNF-α). The pRL was internalized into DCs via a rapid and efficient method. The mice immunized with pRL exhibited enhanced antigen-specific serum IgG and Th1-(IFN-γ), Th2-(IL-4), and Th17-(IL-17, IL-6) cytokine production, contributing to a good anti-infection performance. Furthermore, the pRL could effectively deliver the antigen to the lymph nodes (LNs), activate DC in the LN and produce enhanced CD4+and CD8+ T-cells-derived memory (CD44high CD62Lhigh), and effector (CD44high CD62Llow) as well as functional phenotypes. Our results revealed that pRL can act as a promising adjuvant with targeted delivery of antigen due to its effective activation and robust adaptive immunity induction of DCs.

Glucagon-like peptides (GLP-1 and GLP-2) are two proglucagon-derived intestinal hormones that mediate distinct physiological functions through two related receptors (GLP-1R and GLP-2R) which are important drug targets for metabolic disorders and Crohn’s disease, respectively. Despite great progress in GLP-1R structure determination, our understanding on the differences of peptide binding and signal transduction between these two receptors remains elusive. Here we report the electron microscopy structure of the human GLP-2R in complex with GLP-2 and a Gs heterotrimer. To accommodate GLP-2 rather than GLP-1, GLP-2R fine-tunes the conformations of the extracellular parts of transmembrane helices (TMs) 1, 5, 7 and extracellular loop 1 (ECL1). In contrast to GLP-1, the N-terminal histidine of GLP-2 penetrates into the receptor core with a unique orientation. The middle region of GLP-2 engages with TM1 and TM7 more extensively than with ECL2, and the GLP-2 C-terminus closely attaches to ECL1, which is the most protruded among 9 class B G protein-coupled receptors (GPCRs). Functional studies revealed that the above three segments of GLP-2 are essential for GLP-2 recognition and receptor activation, especially the middle region. These results provide new insights into the molecular basis of ligand specificity in class B GPCRs and may facilitate the development of more specific therapeutics.

Background Recent studies have significantly expanded our knowledge of viral diversity and functions in the environment. Exploring the ecological relationships between viruses, hosts, and the environment is a crucial first step towards a deeper understanding of the complex and dynamic interplays among them. Results Here, we obtained extensive 16S rRNA gene amplicon, metagenomics sequencing, and geochemical datasets from different depths of two highly stratified sulfidic mine tailings cores with steep geochemical gradients especially pH, and explored how variations in viral community composition and functions were coupled to the co-existing prokaryotic assemblages and the varying environmental conditions. Our data showed that many viruses in the mine tailings represented novel genera, based on gene-sharing networks. Siphoviridae, Podoviridae, and Myoviridae dominated the classified viruses in the surface tailings and deeper layers. Both viral richness and normalized coverage increased with depth in the tailings cores and were significantly correlated with geochemical properties, for example, pH. Viral richness was also coupled to prokaryotic richness (Pearson’s r = 0.65, P = 0.032). The enrichment of prophages in the surface mine tailings suggested a preference of lysogenic viral lifestyle in more acidic conditions. Community-wide comparative analyses clearly showed that viruses in the surface tailings encoded genes mostly with unknown functions while viruses in the deeper layers contained genes mainly annotated as conventional functions related to metabolism and structure. Notably, significantly abundant assimilatory sulfate reduction genes were identified from the deeper tailings layers and they were widespread in viruses predicted to infect diverse bacterial phyla. Conclusions Overall, our results revealed a depth-related distribution of viral populations in the extreme and heterogeneous tailings system. The viruses may interact with diverse hosts and dynamic environmental conditions and likely play a role in the functioning of microbial community and modulate sulfur cycles in situ.

Non-obstructive azoospermia (NOA) affects 1% of men. However, the unknowns of NOA pathogenesis and even normal spermatogenic microenvironment establishment severely limit the clinical efficacy of NOA treatment. We profiled > 80,000 human testicular single-cell transcriptomes from 10 healthy donors spanning the range from infant to adult and 7 NOA patients. Sertoli cells, which form the scaffold in the testicular microenvironment, exhibited the most obvious damages in NOA patients. We identified the roadmap of Sertoli cell maturation. Notably, Sertoli cells of patients with congenital causes (Klinefelter syndrome and Y chromosome microdeletions) are mature but with abnormal immune response, while the cells in idiopathic NOA (iNOA) are basically physiologically immature. Furthermore, inhibition of Wnt signaling promotes the maturation of Sertoli cells from iNOA patients, allowing these cells to regain their ability to support germ cell survival. We provide a novel perspective on the development of diagnostic methods and therapeutic targets for NOA.

The yft1 tomato mutant has a yellow-fruited phenotype controlled by a recessive gene (YFT1), which has been shown by map-based cloning to be a homolog of ETHYLENE INSENSITIVE 2 (EIN2). The genetic lesion of the YFT1 allele of yft1 is attributed to a 573 bp DNA fragment (IF573) insertion at 1,200 bp downstream of the transcription start site (TSS). Transcriptome analysis revealed that the mutation resulted in 5,053 differentially expressed genes (DEGs) in the fruit pericarp compared with the M82 wild type cultivar. These were annotated as being involved in ethylene synthesis, chromoplast development, and carotenoid synthesis. Genetic lesion in YFT1 caused a reduction in its own transcript levels in yft1 and impaired ethylene emission and signal transduction, delayed chromoplast development and decreased carotenoid accumulation. The molecular mechanism underlying the reduced expression of YFT1 in yft1 was examined at both the RNA and DNA levels. The IF573 event was shown to introduce two negative regulatory sequences located at -272 to -173 bp and -172 to -73 bp in the YFT1 allele promoter, causing alterative splicing due to aberrant splicing sites, and also altering the structure of the open reading frame in the 5’-UTR. This study contributes to the understanding of color formation in tomato fruit.One-sentence summaryLesion happened in regulatory region impairs expression of a key gene of ethylene signal pathway, and alters fruit color in tomato due to effect of carotenoids accumulation and ethylene synthesis.

2’-Acetylacteoside-(2’-AA), a bioactive constituent isolated from Cistanche deserticola, has been proven to possess a variety of important pharmacological effects, thus brought an increased amount of scientists’ attention. As the extract of C. deserticola exhibited significant anti-osteoporotic bioactivity in our previous study, we proposed that 2’-AA maybe one of the responsibilities. As a result, 2’-AA (10, 20 and 40 mg/kg body weight/day) exhibited significant anti-osteoporotic effects on ovariectomized (OVX) mice after 12 weeks of oral administration, confirmed by the increased bone mineral density, enhanced bone strength and improved trabecular bone micro-architecture including bone mineral content, tissue mineral content, trabecular number, and trabecular separation of OVX mice. Moreover, the properties of bone resorption markers including cathepsin K, TRAP and deoxypyridinoline were significantly suppressed, whereas the activities of bone formation index like ALP and BGP as well as the weights of the body, uterus, and vagina were seemingly not influenced by 2’-AA intervention. Mechanistically, the above therapeutic effect of 2’-AA on bone resorption of OVX mice operated maybe mainly through RANKL/RANK/TRAF6-mediated NF-κB/NFATc1 pathway, which was confirmed by the down-regulated expressions of RANK, TRAF6, IκB kinase β, NF-κB and NFATc1. Summarily, 2’-AA exhibited significant anti-osteoporotic activity and may be regarded as a promising anti-osteoporotic candidate for future clinical trial.

Several studies have demonstrated the effect of probiotics on prevention of diseases in infants. However, the mechanisms remain not fully addressed. In the present study a modified standard ileal efflux medium (SIEM) was exploited in an in vitro analysis and the effect of Lactobacillus plantarum ST-III on the composition of infant gut microbiota was examined in I-chip analysis followed by qPCR confirmation. Also, the effect of ST-III, when administered in combination with breast milk and infant formula, was investigated. The results showed that ST-III could reduce the overall amount of Enterobacteria and Bacteroidetes bacteria, yet increase the abundance of Lactobacillus and certain Bifidobacterium species. Notably, some species/genus of bacteria of medical significance, e.g., Alcaligenaceae sutterella and Desulfovibrio bacteria were inhibited by ST-III. Moreover, co-administration of breast milk or infant formula could increase the abundance of ST-III, suggesting a synergistic effect between ST-III and infant formula and breast milk.

A normalized full-length cDNA library was constructed from the coralloid roots of Cycas debaoensis by the DSN (duplex-specific nuclease) normalization method combined with the SMART (Switching Mechanism At 5′ end of the RNA Transcript) technique. The titer of the original cDNA library was about 1.5 × 106 cfu·mL−1 and the average insertion size was about 1 kb with a high recombination rate (97%). The 5011 high-quality expressed sequence tags (ESTs) were obtained from 5393 randomly picked cDNA clones. Clustering and assembly of ESTs resulted in 2984 unique sequences, consisting of 618 contigs and 2366 singlets. EST sequence annotation revealed that 2333 and 1901 unigenes were functionally annotated in the NCBI non-redundant database and Swiss-Prot protein database, respectively. Functional analysis demonstrated that 1495 (50.1%) unigenes were associated with 4082 Gene Ontology (GO) terms. A total of 847 unigenes were grouped into 22 Cluster of Orthologous Groups (COG) functional categories. Based on the EST dataset, 22 ESTs that encoded putative receptor-like protein kinase (RLK) genes were screened. Furthermore, a total of 94 simple sequence repeats (SSRs) were discovered, of which 20 loci were successfully amplified in C. debaoensis. This study is the first EST analysis for the coralloid roots of C. debaoensis and provides a valuable genomic resource for novel gene discovery, gene expression and comparative genomics, conservation and management studies as well as applications in C. debaoensis and related cycad species. Keywords: Cycas debaoensis, Coralloid root, cDNA library, Expressed sequence tags, Symbiosis and defense, SSRs

Given the limitations of existing therapeutic agents for treatment of postmenopausal osteoporosis, there still remains a need for more options with both efficacy and less adverse effects. Cistanche deserticola Y. C. Ma is known as a popular tonic herb traditionally used to treatment deficiency of kidney energy including muscle weakness in minority area of Asian counties. Based on the theory of "kidney dominate bone," an ovariectomized (OVX) rat model of postmenopausal osteoporosis was used to evaluate the therapeutic effect of C. deserticola extract (CDE) on bone loss. Forty eight female Sprague-Dawley rats, aged about 12 weeks, were randomly assigned into six groups including sham group orally administrated with 0.5% carboxymethyl cellulose sodium (CMC-Na) (sham), positive group treated with 1 mg/kg of estradiol valerate (EV), low, moderate, and high dosage groups orally administrated with 200, 400, and 800 mg/kg/day of CDE, respectively. After 3 months of continuous intervention, CDE exhibited significant anti-osteoporotic activity evidenced by the enhanced total bone mineral density, ameliorated bone microarchitecture; increased alkaline phosphatase activity; decreased deoxypyridinoline, cathepsin K, tartrate-resistant acid phosphatase, and malondialdehyde levels; whereas the body, uterus, and vagina weights in OVX rats were not influenced by CDE intervention. In addition, a seemed contradictory phenomenon on levels of calcium and phosphorus between OVX and sham rats were observed and elucidated. Mechanistically, CDE significantly down-regulated the levels of TRAF6, RANKL, RANK, NF-κB, IKKβ, NFAT2, and up-regulated the phosphatidylinositol 3-kinase (PI3K), AKT, osteoprotegerin, and c-Fos expressions, which implied CDE could suppress RANKL/RANK-induced activation of downstream NF-κB and PI3K/AKT pathways, and ultimately, preventing activity of the key osteoclastogenic proteins NFAT2 and c-Fos. All of the data suggested CDE possessed potential anti-osteoporotic activity and this effect was, at least in part, involved in modulation of RANKL/RANK/TRAF6-mediated NF-κB and PI3K/AKT signaling as well as c-Fos and NFAT2 levels. Therefore, CDE may represent a useful promising remedy candidate for treatment of postmenopausal osteoporosis.

Despite the severity of osteoarthritis (OA), current medical therapy strategies for OA aim at symptom control and pain reduction, as there is no ideal drug for effective OA treatment. OA rat model was used to explore the therapeutic function of quercetin on remission of OA, by determining the reactive oxygen species (ROS) levels, mitochondrial function and extracellular matrix integrity. Quercetin could attenuate ROS generation and augment the glutathione (GSH) and glutathione peroxidase (GPx) expression levels in OA rat. Quercetin not only enhanced mitochondrial membrane potential, oxygen consumption, adenosine triphosphate (ATP) levels in mitochondria, but also increased the mitochondrial copy number. Furthermore, the interlukin (IL)-1β-induced accumulation of nitric oxide (NO), matrixmetalloproteinase (MMP)-3) and MMP-13 could be suppressed by quercetin. Finally, we confirmed that the therapeutic properties of quercetin on OA might function through the adenosine monophosphate-activated protein kinase/sirtuin 1 (AMPK/SIRT1) signaling pathway. In summary, quercetin could alleviate OA through attenuating the ROS levels, reversing the mitochondrial dysfunction and keeping the integrality of extracellular matrix of joint cartilage. The underlying mechanism might involve the regulation of AMPK/SIRT1 signaling pathway.

The aim of the study was to elucidate the therapeutic effects of Cytisine (CYT) on cerebral ischemia-reperfusion injury in mice. Male ICR mice were pretreated with reagents (drug), and then subjected to 2 h focal cerebral ischemia and 24 h reperfusion. Morphologically, the histopathological impairment were estimated by the TTC, HE and TUNEL staining. The expression of GluN2B-containing NMDA receptor, phosphorylation of extracellular regulated protein kinases, total ERK, phosphorylation of cAMP-response element binding protein and total CREB were determined by immunofluorescence and Western blot assay, respectively. The mRNA expression of NR2B, ERK and CREB were quantified by the real-time RT-PCR. CYT significantly diminished the infarct size and neuronal apoptosis. Additionally, it ameliorated histopathological lesion dramatically. CYT promoted the phosphorylation of ERK, CREB and their mRNA expression. In contrast, the expression of NR2B was suppressed in concomitant with the down-regulation of genes. The overall results thus far suggest that CYT confers the neuroprotection against cerebral I/R injury by regulating the NR2B-ERK/CREB signal pathway.

Flowering time is a trait vital to the adaptation of flowering plants to different environments. Here, we report that CCT domain genes play an important role in flowering in maize (Zea mays L.). Among the 53 CCT family genes we identified in maize, 28 were located in flowering time quantitative trait locus regions and 15 were significantly associated with flowering time, based on candidate-gene association mapping analysis. Furthermore, a CCT gene named ZmCOL3 was shown to be a repressor of flowering. Overexpressing ZmCOL3 delayed flowering time by approximately 4 d, in either long-day or short-day conditions. The absence of one cytosine in the ZmCOL3 3'UTR and the presence of a 551 bp fragment in the promoter region are likely the causal polymorphisms contributing to the maize adaptation from tropical to temperate regions. We propose a modified model of the maize photoperiod pathway, wherein ZmCOL3 acts as an inhibitor of flowering either by transactivating transcription of ZmCCT, one of the key genes regulating maize flowering, or by interfering with the circadian clock.