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The aim of the study was to investigate the role of NSUN2 (NOP2/Sun RNA Methyltransferase Family Member 2) in hepatocellular carcinoma (HCC). The expressions of NSUN2 and FZR1 were measured. Cell viability, proliferation, and apoptosis were assessed. HCC xenograft in nude mouse model was established. Tumor weight and volume were examined. Tumor tissues were collected for immunohistochemistry (IHC). TCGA database analysis and clinical sample testing suggested that the transcript levels of NSUN2 and FZR1 were increased in HCC tissues. NSUN2 knockdown inhibited HCC cell viability and proliferation, and promoted cell apoptosis. Moreover, the effects of NSUN2 could be countered by overexpressing FZR1. In animal experiment, NSUN2 silencing suppressed tumor growth in nude mice by downregulating FZR1. In conclusion, NSUN2 has a regulatory effect on HCC cell proliferation and apoptosis. NSUN2 knockout could inhibit cellular processes in HCC and tumor growth, likely via FZR1 inhibition. This finding has not only revealed the role of NSUN2 in HCC growth, but also suggests a promising target for HCC treatment.

Hexokinase (HXK) is the first irreversible catalytic enzyme in the glycolytic pathway, which not only provides energy for plant growth and development but also serves as a signaling molecule in response to environmental changes. However, the evolutionary pattern of the HXK gene family in apple remains unknown. In this study, a total of nine HXK genes were identified in the Malus×domestica genome GDDH13 v1.1. The physiological and biochemical properties, exonintron structures, conserved motifs, and cis-elements of the MdHXK genes were determined. Predicted subcellular localization indicated that the MdHXK genes were mainly distributed in the mitochondria, cytoplasm, and nucleus. Gene duplication revealed that whole-genome duplication (WGD) and segmental duplication played vital roles in MdHXK gene family expansion. The ω values of pairwise MdHXK genes indicated that this family was subjected to strong purifying selection during apple domestication. Additionally, five subfamilies were classified, and recent/old duplication events were identified based on phylogenetic tree analysis. Different evolutionary rates were estimated among the various HXK subfamilies. Moreover, divergent expression patterns of the MdHXK genes in four source-sink tissues and at five different apple fruit developmental stages indicated that they play vital roles in apple fruit development and sugar accumulation. Our study provides a theoretical basis for future elucidation of the biological functions of the MdHXK genes during apple fruit development.

Background & aims Some patients with SARS-CoV-2 infection have abnormal liver function. We aimed to clarify the features of COVID-19-related liver damage to provide references for clinical treatment. Methods We performed a retrospective, single-center study of 148 consecutive patients with confirmed COVID-19 (73 female, 75 male; mean age, 50 years) at the Shanghai Public Health Clinical Center from January 20 through January 31, 2020. Patient outcomes were followed until February 19, 2020. Patients were analyzed for clinical features, laboratory parameters (including liver function tests), medications, and length of hospital stay. Abnormal liver function was defined as increased levels of alanine and aspartate aminotransferase, gamma glutamyltransferase, alkaline phosphatase, and total bilirubin. Results Fifty-five patients (37.2%) had abnormal liver function at hospital admission; 14.5% of these patients had high fever (14.5%), compared with 4.3% of patients with normal liver function (P = .027). Patients with abnormal liver function were more likely to be male, and had higher levels of procalcitonin and C-reactive protein. There was no statistical difference between groups in medications taken before hospitalization; a significantly higher proportion of patients with abnormal liver function (57.8%) had received lopinavir/ritonavir after admission compared to patients with normal liver function (31.3%). Patients with abnormal liver function had longer mean hospital stays (15.09 ± 4.79 days) than patients with normal liver function (12.76 ± 4.14 days) (P = .021). Conclusions More than one third of patients admitted to the hospital with SARS-CoV-2 infection have abnormal liver function, and this is associated with longer hospital stay. A significantly higher proportion of patients with abnormal liver function had received lopinavir/ritonavir after admission; these drugs should be given with caution.

The rhythmic regulation of transcriptional processes is intimately linked to lipid homeostasis, to anticipate daily changes in energy access. The Rev-erbα-HDAC3 complex was previously discovered to execute the rhythmic repression of lipid genes; however, the epigenetic switch that turns on these genes is less clear. Here, we show that genomic recruitment of MRG15, which is encoded by the mortality factor on chromosome 4 (MORF4)-related gene on chromosome 15, displays a significant diurnal rhythm and activates lipid genes in the mouse liver. RNA polymerase II (Pol II) recruitment and histone acetylation correspond to MRG15 binding, and the rhythm is impaired upon MRG15 depletion, establishing MRG15 as a key modulator in global rhythmic transcriptional regulation. MRG15 interacts with the nuclear receptor LRH-1, rather than with known core clock proteins, and is recruited to genomic loci near lipid genes via LRH-1. Blocking of MRG15 by CRISPR targeting or by the FDA-approved drug argatroban, which is an antagonist to MRG15, attenuates liver steatosis. This work highlights MRG15 as a targetable master regulator in the rhythmic regulation of hepatic lipid metabolism.

SummaryCaspase recruitment domain containing protein 9 (CARD9) is an adaptor protein that plays a critical role in pattern recognition receptors (PRRs)-mediated activation of NF-?B and mitogen-activated protein kinase (MAPK). This elicits initiation of the pro-inflammatory cytokines and leads to inflammatory responses, which has been recognized as a critical contributor to chronic inflammation. Current researches demonstrate that CARD9 is strongly associated with metabolic diseases, such as obesity, insulin resistance, atherosclerosis and so on. In this review, we summarize CARD9 signaling pathway and the role of CARD9 in metabolic diseases.

Methionine enkephalin (MENK), an endogenous opioid peptide, has a role in nervous system, immune system, and anticancer therapy. Inflammation, metabolism and cancer are closely intertwined with each other. This study is to identify the correlation of the antitumor effects of MENK with systemic inflammation, liver metabolism, and immune cells as myeloid-derived suppressor cells (MDSCs). We established a subcutaneous CT26 colon carcinoma model and a cyclophosphamide-induced immunosuppressive model subjected to MENK. AML12 and MDSCs were used as in vitro models. The results showed that MENK treatment degraded tumor growth and inhibited proinflammatory cytokines both in tumor tissues and serum. The MENK-treated tumor mice showed normalized liver function with glycolipid metabolic homeostasis. No inhibitory effect on CT26 tumor cell in vitro, but only reduced lipid synthesis in AML12 were presented by MENK. Meanwhile, MENK invigorated immune response in both two animal models by markedly suppressing MDSCs and enhancing T cells response. In vitro MENK-treated MDSCs showed reduced glycolysis and less ROS production, which was mediated by PI3K/AKT/mTOR pathway. Opioid receptor antagonist naltrexone reversed most of the regulation. These results illustrate that MENK preventing development of colon carcinoma might be correlated with the suppression of inflammation, improving metabolism in liver as well as in MDSCs partly through opioid receptor, which brings new elements supporting the adjuvant therapy for tumor by MENK.

Background and RationaleHepatoblastoma (HB) is the most common pediatric liver cancer. Its predominant occurrence in very young children led us to investigating whether the neonatal liver provides a protumorigenic niche to HB development.MethodsHB development was compared between orthotopic transplantation models established in postnatal day 5 and 60 mice (P5Tx and P60Tx models). Single-cell RNA-sequencing was performed using tumor and liver tissues from both models and the top candidate cell types and genes identified are investigated for their roles in HB cell growth, migration, and survival.ResultsWe found that various HB cell lines including HepG2 cells were consistently and considerably more tumorigenic and metastatic in the P5Tx model than in the P60Tx models. Sc-RNAseq of the P5Tx and P60Tx HepG2 models revealed that the P5Tx tumor was more hypoxic and had a larger number of activated hepatic stellate cells (aHSCs) in the tumor-surrounding liver which express significantly higher levels of Cxcl1 than those from the P60Tx model. We found these differences were developmentally present in normal P5 and P60 liver. We showed that the Cxcl1/Cxcr2 axis mediated HB cell migration and was critical to HB cell survival under hypoxia. Treating HepG2 P60Tx model with recombinant CXCL1 protein induced intrahepatic and pulmonary metastasis and CXCR2 knockout in HepG2 cells abolished their metastatic potential in the P5Tx model. Lastly, we showed that in metastatic HB patient tumors there was a similar larger population of aHSCs in the tumor-surrounding liver than in localized tumors, and tumor hypoxia was uniquely associated with HB patient prognosis among pediatric cancers.ConclusionWe demonstrated that the neonatal liver provides a prometastatic niche to HB development via the Cxcl1/Cxcr2 axis.

Accumulated evidence shows that complex microbial communities resides in the healthy human urinary tract and can change in urological disorders. However, there lacks a comprehensive profiling of the genitourinary microbiota in healthy cohort. Here, we performed 16S rRNA gene sequencing of midstream urine specimens from 1,172 middle-aged and elderly healthy individuals. The core microbiota included 6 dominant genera (mean relative abundance >5%), including Prevotella, Streptococcus, Lactobacillus, Gardnerella, Escherichia-Shigella, and Veillonella, and 131 low-abundance genera (0.01–5%), displaying a distinct microbiome profiles to that of host-matched gut microbiota. The composition and diversity of genitourinary microbiome (GM) were distinct between genders and may fluctuate with ages. Several urotypes were identified by the stratification of microbiome profiles, which were mainly dominated by the six most predominant genera. The prevalence of urotypes was disparate between genders, and the male sample additionally harbored other urotypes dominated by Acinetobacter, Corynebacterium, Staphylococcus, or Sphingomonas. Peptoniphilus, Ezakiella, and Porphyromonas were co-occurred and co-abundant, and they may play crucial roles as keystone genera and be associated with increased microbial diversity. Our results delineated the microbial structure and diversity landscape of the GM in healthy middle-aged and elderly adults and provided insights into the influence of gender and age to it.

Radiotherapy represents the most effective nonsurgical therapy, whereas acquired radioresistance remains a major challenge in glioma treatment. Deregulation of long noncoding RNAs (lncRNAs) is frequently involved in tumorigenesis. This study investigates the role of LINC01123 in radioresistance in glioma with molecules involved. LINC01123 was identified as the most upregulated gene in a glioma gene expression dataset GSE103227. LINC01123 was highly expressed in the radioresistant glioma tissues radioresistant glioma U251 (U251R) cells. Downregulation of LINC01123 reduced cell proliferation and colony formation abilities, as well as resistance to apoptosis of the U251R cells after 4 Gy X-ray irradiation. The micro(mi)RNA-151a gene (miR-151a) was a poorly expressed miRNA in glioma, and it was a target of LINC01123. The centromere protein B gene (CENPB) mRNA was a direct target of miR-151a and demonstrated a positive correlation with LINC01123 in glioma tissues and cells. Further inhibition of miR-151a or overexpression of CENPB restored radioresistance of glioma cells. In addition, silencing of LINC01123 suppressed growth of xenograft tumors formed by U251R cells in nude mice. To conclude, the present study demonstrates that LINC01123 serves as a sponge for miR-151a and upregulates CENPB expression to increase the radioresistance of glioma cells in vitro and in vivo.

Formed by back splicing or back fusion of linear RNAs, circular RNAs (circRNAs) constitute a new class of non-coding RNAs of eukaryotes. Recent studies reveal a spliceosome-dependent biogenesis of circRNAs where circRNAs arise at the intron-exon junctions of mRNAs. In this study, using a novel de novo identification method, we show that circRNAs can originate from the interior regions of exons, introns, and intergenic transcripts in human, mouse and rice, which were referred to as interior circRNAs (i-circRNAs). Many i-circRNAs have some remarkable characteristics: multiple i-circRNAs may arise from the same genomic locus; their back fusion points may not be associated with the AG/GT splicing sites, but rather a new pair of motif AC/CT, their back fusion points are adjacent to complementary sequences; and they may circulate on short homologous sequences. We validated several i-circRNAs in HeLa cells by Polymerase Chain Reaction followed by Sanger sequencing. Our results combined showed that i-circRNAs are bona fide circRNAs, indicated novel biogenesis pathways independent of the splicing apparatus, and expanded our understanding of the origin, diversity, and complexity of circRNAs.

Ginsenoside Rh2 (G-Rh2) has well-established potent antitumor activity; yet, the effects of G-Rh2 on immune and metabolism regulation in cancer treatment, especially non-small cell lung cancer (NSCLC) remain unclear. We showed that G-Rh2 had a synergistic antitumor effect with cyclophosphamide (CY) on mice with NSCLC, and improved the immune deficiency caused by CY. Consistently, G-Rh2 exhibited no inhibitory effect on tumor growth of T cells-deficient nude mice. Furthermore, G-Rh2 treatment triggered the oxidative decomposition of fatty acid (FA), suppressed FA synthesis, increased ketone level, and decreased glucocorticoid (CORT) secretion. G-Rh2 significantly down-regulated the expression of fatty acid synthase (FASN). Of note, in liver-specific FASN knockout mice G-Rh2 failed to show the same immune enhancement effects. Further mechanistic exploration revealed that G-Rh2 suppressed the expression and nuclear translocation of sterol regulatory element binding protein 1 (SREBP-1), and disturbed the SREBP-1–FASN interaction in vitro.

The diversity of the bacterial community in the gut is closely related to human health. Gut microbes accomplish multiple physiological and biochemical functions. Sitosterols are a series of phytochemicals that have multiple pharmacological activities and are used as cholesterol-lowering drugs in clinical practice. In this study, we investigated the roles of bacteria and short-chain fatty acids (SCFAs) to the anti-colorectal cancer (anti-CRC) effects of sitosterols in BALB/c nude mice. Sitosterols were administered orally and gut microbiota composition and intestinal SCFAs changes were analyzed. The correlation between gut microbiota, SCFAs, and tumor apoptosis was assessed by a series of in vivo and in vitro experiments. Tumor growth in the mice was inhibited by sitosterol-treatment. Mechanistic studies revealed that sitosterol-treatment reduced the expression of PI3K/Akt, promoted the activation of Bad, decreased Bcl-xl, and enhanced cyto-c release, leading to caspase-9 and caspase-3 activation, PARP cleavage, and apoptosis. 16S rDNA analysis revealed that the diversity of microbiota, particularly phyla Bacteroidetes and Firmicutes, reduced dramatically in the gut of tumor-bearing mice, whilst treatment with sitosterols reversed these changes. The levels of SCFAs in the fecal samples of sitosterol-treated mice increased, leading to cancer cell apoptosis in vitro. Moreover, tumor apoptosis was induced after mice received a daily dose of 2 × 108 CFU/0.2 mL Lactobacillus pentosus or 20 mM/0.2 mL SCFAs. Taken together, these results demonstrate that sitosterols maintain a diverse microbial environment and enrich the content of L. pentosus in the gut, leading to the production of beneficial metabolites including SCFAs that promote tumor apoptosis.

A better understanding of the molecular mechanisms that control the UCP1 expression in brown and beige adipocytes is essential for us to modulate adipose cell fate and promote thermogenesis, which may provide a therapeutic view for the treatment of obesity and obesity-related diseases. To systematically identify cis-element(s) that transcriptionally regulates Ucp1, we here took advantage of the high-throughput CRIPSR-Cas9 screening system, and performed an in situ saturating mutagenesis screen, by using a customized sgRNA library targeting the ~ 20 kb genomic region near Ucp1. Through the screening, we have identified several genomic loci that may contain key regulatory element for Ucp1 expression in cultured brown and white adipocytes in vitro, and in inguinal white adipose tissue in vivo. Our study highlights a broadly useful approach for studying cis-regulatory elements in a high-throughput manner. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s43657-020-00006-7) contains supplementary material, which is available to authorized users.

The woman’s gut microbiota during pregnancy may support nutrient acquisition, is associated with diseases, and has been linked to infant health. However, there is limited information on gut microbial characteristics and dependence in pregnant women. In this study, we provide a comprehensive overview of the gut microbial characteristics of 1479 pregnant women using 16S rRNA gene sequencing of fecal samples. We identify a core microbiota of pregnant women, which displays a similar overall structure to that of age-matched nonpregnant women. Our data show that the gestational age-associated variation in the gut microbiota, from the ninth week of gestation to antepartum, is relatively limited. Building upon rich metadata, we reveal a set of exogenous and intrinsic host factors that are highly correlated with the variation in gut microbial community composition and function. These microbiota covariates are concentrated in basic host properties (e.g., age and residency status) and blood clinical parameters, suggesting that individual heterogeneity is the major force shaping the gut microbiome during pregnancy. Moreover, we identify microbial and functional markers that are associated with age, pre-pregnancy body mass index, residency status, and pre-pregnancy and gestational diseases. The gut microbiota during pregnancy is also different between women with high or low gestational weight gain. Our study demonstrates the structure, gestational age-associated variation, and associations with host factors of the gut microbiota during pregnancy and strengthens the understanding of microbe–host interactions. The results from this study offer new materials and prospects for gut microbiome research in clinical and diagnostic fields.

Understanding activity patterns of large mammals provides a lens to view potential temporal niches, while studies on daily activity patterns are helpful to understand behavioral ecology of large mammals and their conservation. We studied the seasonal daily activity patterns and effects of livestock grazing on three large mammals: giant panda, tufted deer, and Chinese serow in Wanglang National Nature Reserve, Sichuan Province, China. The comparison of seasonal daily activity patterns indicated that the giant panda exhibited a single daily peak, whereas tufted deer exhibited two and Chinese serow exhibited multiple peaks. All three large mammals differed significantly in daily activity patterns, while only tufted deer and Chinese serow showed moderate overlap. Livestock grazing (an example of human disturbance) showed high overlap coefficients in daily activity patterns with three mammals, while there was no significant difference between giant pandas and livestock. These results provided a critical baseline mechanism for species coexistence that can help in applied efforts for conservation purposes such as habitat management. In addition, some suggestions for livestock management and disturbance pattern analysis may provide new insights into threatened species conservation.

While Baccillus Calmette-Guerin (BCG) is used worldwide, tuberculosis (TB) is still a global concern due to the poor efficacy of BCG. Novel vaccine candidates are therefore urgently required. In this study, two attenuated recombinant Listeria strains, LMΔ-msv and LIΔ-msv were constructed by deletion of actA and plcB and expression of a fusion protein consisting of T cell epitopes from four Mycobacterium tuberculosis (Mtb) antigens (Rv2460c, Rv2660c, Rv3875, and Rv3804c). The safety and immunogenicity of the two recombinant strains were evaluated in C57BL/6J mice. After intravenous immunization individually, both recombinant strains entered liver and spleen but eventually were eliminated from these organs after several days. Simultaneously, they induced antigen-specific cell-mediated immunity, indicating that the recombinant Listeria strains were immunogenic and safe in vivo. LMΔ-msv immunization induced stronger cellular immune responses than LIΔ-msv immunization, and when boosted with LIΔ-msv, antigen-specific IFN-γ CD8+ T cell responses were notably magnified. Furthermore, we evaluated the protection conferred by the vaccine candidates against mycobacterial infection via challenging the mice with 1 × 107 CFU of BCG. Especially, we tested the feasibility of application of them as heterologous BCG supplement vaccine by immunization of mice with BCG firstly, and boosted with LMΔ-msv and LIΔ-msv sequentially before challenging. Combination immune strategy (LMΔ-msv prime-LIΔ-msv boost) conferred comparable protection efficacy as BCG alone. More importantly, BCG-vaccinated mice acquired stronger resistance to Mycobacterial challenge when boosted with LMΔ-msv and LIΔ-msv sequentially. Our results inferred that heterologous immunization with Listeria-based recombinant strains boosted BCG-primed protection against pulmonary mycobacterial infection.

Objective To investigate the expression of aquaporin 7 (AQP7) and aquaporin 9 (AQP9) in the granulosa cells of patients with polycystic ovary syndrome (PCOS) and healthy women and detect their localization in oocytes at the germinal vesicle (GV), metaphase I (MI), MII, embryo, and blastocyst stages and the in vitro response to insulin stimulation. Design Randomized, assessor-blinded study. Setting Reproductive medical center. Patient(s) A total of 40 women (aged 20–38 years) comprising 29 cases of primary infertility and 11 cases of secondary infertility, of whom 17 had an initial diagnosis of PCOS and three received a PCOS diagnosis after an infertility examination. Intervention(s) Controlling different concentrations of insulin and different treatment times in cultures of normal human granulosa cells in vitro. Main Outcome Measure(s) Expression of AQP7 and AQP9 genes and proteins in granulosa cells detected by real-time quantitative polymerase chain reaction, and localization in oocytes at the GV, MI, MII, embryo, and blastocyst stages by Western blot, immunohistochemical, and immunofluorescence assays, and concentrations of insulin in follicular fluid by enzyme-linked immunosorbent assay. Result(s) The expression levels of the AQP7 mRNA and protein in the granulosa cells of patients with PCOS were higher than found in healthy controls. We found AQP7 protein expressed in human oocytes at GV, MI, MII, embryo, and blastocyst stages; it was mainly located in the nucleoplasm. In the PCOS group, the expression level of AQP9 mRNA and protein in granulosa cells was lower, and AQP9 protein was expressed in oocytes at the GV, MI, MII, embryo, and blastocyst stages; it was localized on the nuclear membrane. Compared with healthy women, the insulin expression in patients with PCOS was higher. In cultures of normal human granulosa cells in vitro, the expression of AQP7 and AQP9 mRNA and protein decreased with the increase in insulin concentration; expression statistically significantly decreased when the insulin concentration was 100 nmol/L, and after 6 to 24 hours of exposure the lowest expression levels were found at 12 hours. Conclusion(s) The different localization and expression of AQP7 and AQP9 between the two groups suggests that they might be involved in oocyte maturation and embryonic development through different regulatory pathways. The expression levels of AQP7 and AQP9 were negatively correlated with insulin regulation, suggesting that insulin might affect the maturation of PCOS follicles by changing AQP7 and AQP9 expression.

Cells have developed regulatory mechanisms that underlie flagellar assembly and maintenance, including the transcriptional regulation of flagellar genes, an initial step for making flagella. Although transcriptional regulation of flagellar gene expression is required for flagellar assembly in Chlamydomonas, no transcription factor that regulates the transcription of flagellar genes has been identified. We report that X chromosome-associated protein 5 (XAP5) acts as a transcription factor to regulate flagellar assembly in Chlamydomonas While XAP5 proteins are evolutionarily conserved across diverse organisms and play vital roles in diverse biological processes, nothing is known about the biochemical function of any member of this important protein family. Our data show that loss of XAP5 leads to defects in flagellar assembly. Posttranslational modifications of XAP5 track flagellar length during flagellar assembly, suggesting that cells possess a feedback system that modulates modifications to XAP5. Notably, XAP5 regulates flagellar gene expression via directly binding to a motif containing a CTGGGGTG-core. Furthermore, recruitment of RNA polymerase II (Pol II) machinery for transcriptional activation depends on the activities of XAP5. Our data demonstrate that, through recruitment of Pol II, XAP5 defines a class of transcription factors for transcriptional regulation of ciliary genes. This work provides insights into the biochemical function of the XAP5 family and the fundamental biology of the flagellar assembly, which enhance our understanding of the signaling and functions of flagella.

Lung cancer is the deadliest cancer in the world. Angiogenesis plays a crucial role of the incidence, progression, and metastasis in lung cancer. Angiogenesis inhibitors are used to treat non-small cell lung cancer (NSCLC) patients, and the molecular biomarkers are also being assessed to predict treatment response/therapeutic response and patients' prognosis. Vascular endothelial growth factor (VEGF) is a signal protein produced by cells that stimulates angiogenesis. Due to its predictive values of prognosis on NSCLC, a large number of methods have been developed and evaluated to detect VEGF levels in a variety of studies. In this article, we review the detection methods designed to measure the VEGF levels in different body fluids and prognosticate the value of VEGF in treatment, diagnosis and survival in lung cancer.

This study is aimed at separating and identifying a new glycoprotein from sweet potato (SPG) and detecting its anti-colon cancer bioactivity. The SPG-56 was obtained by water extraction, isoelectric precipitation, chromatography and electrophoresis. The compositions of sugar and protein in SPG-56 were identified with high performance liquid chromatography and amino acid analyzer, and the antitumor activities were explored using BALB/c nude mice and human colon cancer cells (HCT-116). Results showed that the molecular weight of SPG-56 is 56 KD. Relative sugar and protein contents of SPG-56 are 2.9% and 97.1%, comprising more than 6 types of sugars and 15 kinds of amino acids. Animal experiments showed that orally administrated SPG-56 could obviously suppress the tumor growth. Results from flow cytometry, western blotting and immunohistochemistry indicated that SPG-56 inhibits cancer by inducing apoptosis. These results suggest that SPG-56 would potentially delay the development of colon cancer.

Osteoarthritis (OA), which is characterized mainly by cartilage degradation, is the most prevalent joint disorder worldwide. Although OA is identified as a major cause of joint pain, disability, and socioeconomic burden, the etiology of OA is still not clearly known. Recently, gene microarray analysis has become an efficient method for the research of complex diseases and has been employed to determine what genes and pathways are involved in the pathological process of OA. In this review, OA study results over the last decade are summarized for gene expression profiling of various tissues, such as cartilage, subchondral bone, and synovium in human OA and mouse OA models. Many differentially expressed genes, which mainly involve matrix metabolism, bone turnover, and inflammation pathways, were identified in diseased compared with “normal” tissues. Nevertheless, rare common genes were reported from studies using different tissue sources, microarray chips, and research designs. Thus, future novel and carefully designed microarray studies are required to elucidate underlying genetic mechanisms in the pathogenesis of OA as well as new directions for potential OA-targeted pharmaceutical therapies.

As a critically endangered species, Acer catalpifolium Rehd. is included in the wild plants with extremely small populations (WPESP) rescue and protection plan. Understanding of the responses of A. catalpifolium to various resource and environment conditions could help identify potentially suitable environments for its conservation. We used an orthogonal experiment design to evaluate the growth, morphological and physiological responses of A. catalpifolium and a widespread same-genus species A. mono to diverse water and light stresses. The results showed that a 75% soil water holding capacity and moderate shade are suitable for seedling growth. Drought and shade had significant negative effects on plant growth, biomass accumulation, and physiological functions. Lowering specific leaf areas (SLA), increasing root biomass allocation, and pigment content were the main strategies for enhancing water and light capturing abilities to acclimate to the limited ambient resources. The general plasticity index of plant growth, biomass, and physiological characteristics were higher in A. mono than in A. catalpifolium. We also explored the suitable light and water requirements for A. catalpifolium seedling growth. Our research would be useful for population regeneration and could aid in biodiversity conservation of Acer species. Keywords: Acer catalpifolium Rehd, Drought, Shade, Wild plants with extremely small populations (WPESP)

Background: Imbalances in gut microbiota composition are linked to hypertension, host metabolic abnormalities, systemic inflammation, and other conditions. In the present study, we examined the changes of gut microbiota in women with early-onset preeclampsia (PE) and in normotensive, uncomplicated pregnant women during late pregnancy and at 1 and 6 weeks postpartum. Methods: Gut microbiota profiles of women with PE and healthy pregnant women in the third trimester and at 1 and 6 weeks postpartum were assessed by 16S rRNA gene amplicon sequencing. Plasma levels of interleukin-6 (IL-6), intestinal fatty acid-binding protein (I-FABP), zonulin, and lipopolysaccharide (LPS) were measured in the third trimesters. Results: At the genus level, 8 bacterial genera were significantly enriched in the antepartum samples of PE patients compared to healthy controls, of which Blautia, Ruminococcus2, Bilophila, and Fusobacterium represented the major variances in PE microbiomes. Conversely, 5 genera, including Faecalibacterium, Gemmiger, Akkermansia, Dialister, and Methanobrevibacter, were significantly depleted in antepartum PE samples. Maternal blood pressure and liver enzyme levels were positively correlated to the PE-enriched genera such as Anaerococcus, Ruminococcus2, Oribacterium, and Bilophila, while the fetal features (e.g., Apgar score and newborn birth weight) were positively correlated with PE-depleted genera and negatively correlated with PE-enriched genera. Moreover, maternal blood IL-6 level was positively associated with gut Bilophila and Oribacterium, whereas LPS level was negatively associated with Akkermansia. In terms of postpartum women, both the gut microbial composition and the PE-associated microbial alterations were highly consistent with those of the antepartum women. Conclusion: PE diagnosed in the third trimester of pregnancy is associated with a disrupted gut microbiota composition compared with uncomplicated pregnant women, which are associated with maternal clinical features (blood pressure level and liver dysfunction) and newborn birth weight. Moreover, these antepartum alterations in gut microbiota persisted 6 weeks postpartum.

Vitamin C (VC) and vitamin D (VD) have been widely used as the dietary supplements and in treatment of diseases both independently and in combination. Whether there is a connection between their pathways is critical for their therapeutic applications. Using whole-genome expression profiles, we performed multiple measures of associations, networks, eQTL mappings and expressions of key genes of interest in VC and VD functions. Several key genes in their pathways were found to be associated. Gc and Rgn play important roles connecting VC and VD pathways in mice. The r values of expression levels between Gc and Rgn in mouse spleen, liver, lung, and kidney are 0.937, 0.558, 0.901, and 0.617, respectively. The expression QTLs of Gc and Rgn are mapped onto the same locations, i.e., 68–76 MB in chromosome 7 and 26–36 MB in chromosome 9. In humans, there are positive correlations between CYP27B1 and SLC23A1 expression levels in kidney (r = 0.733) and spleen (r = 0.424). SLC23A2 and RXRA are minimally associated in both mouse and human. These data indicate that pathways of VC and VD are not independent but affect each other, and this effect is different between mice and humans during VC and VD synthesis and transportation.

Tet-eleven translocation 1 (TET1) is a dioxygenase that plays an important role in decreasing the abundance of DNA methylation and changing the expression levels of specific genes related to inflammation. Porphyromonas gingivalis (Pg.) lipopolysaccharide (LPS) can induce periodontal diseases that present with severe bone loss and collagen fiber destruction accompanied by a high number of M1 macrophages. M1-polarized macrophages are pivotal immune cells that promote the progression of the periodontal inflammatory response, but the function of TET1 during M1 macrophage activation is still unknown. Our results showed that the mRNA and protein expression levels of TET1 decreased in THP-1 cells during M1 macrophage differentiation. TET1 knockdown resulted in a significant decrease in the production of proinflammatory markers such as IL-6, TNF-&alpha, CCL2, and HLA-DR in Pg. LPS/IFN-&gamma, and Escherichia coli (E. coli) LPS/IFN-&gamma, induced M1 macrophages. Mechanistically, TET1 knockdown downregulated the activity of the NF-&kappa, B signaling pathway. After treatment with the NF-&kappa, B inhibitor BAY 11-7082, M1 marker expression showed no significant difference between the TET1 knockdown group and the control group. Taken together, these results suggest that TET1 depletion inhibited Pg. LPS/IFN-&gamma, induced M1 macrophage polarization through the NF-&kappa, B pathway in THP-1 cells.

Bone mesenchymal stem cells (BMSCs) can be a useful cell resource for developing biological treatment strategies for bone repair and regeneration, and their therapeutic applications hinge on an understanding of their physiological characteristics. N6-methyl-adenosine (m6A) is the most prevalent internal chemical modification of mRNAs and has recently been reported to play important roles in cell lineage differentiation and development. However, little is known about the role of m6A modification in the cell differentiation of BMSCs. To address this issue, we investigated the expression of N6-adenosine methyltransferases (Mettl3 and Mettl14) and demethylases (Fto and Alkbh5) and found that Mettl3 was upregulated in BMSCs undergoing osteogenic induction. Furthermore, we knocked down Mettl3 and demonstrated that Mettl3 knockdown decreased the expression of bone formation-related genes, such as Runx2 and Osterix. The alkaline phosphatase (ALP) activity and the formation of mineralized nodules also decreased after Mettl3 knockdown. RNA sequencing analysis revealed that a vast number of genes affected by Mettl3 knockdown were associated with osteogenic differentiation and bone mineralization. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis revealed that the phosphatidylinositol 3-kinase/AKT (PI3K-Akt) signaling pathway appeared to be one of the most enriched pathways, and Western blotting results showed that Akt phosphorylation was significantly reduced after Mettl3 knockdown. Mettl3 has been reported to play an important role in regulating alternative splicing of mRNA in previous research. In this study, we found that Mettl3 knockdown not only reduced the expression of Vegfa but also decreased the level of its splice variants, vegfa-164 and vegfa-188, in Mettl3-deficient BMSCs. These findings might contribute to novel progress in understanding the role of epitranscriptomic regulation in the osteogenic differentiation of BMSCs and provide a promising perspective for new therapeutic strategies for bone regeneration.

Understanding the relationships between giant panda (Ailuropoda melanoleuca) and their sympatric species is an important component in their conservation. In this study we evaluate the impact of sympatric species with three ranges of body size on the distribution and habitat selection of giant panda. We surveyed giant panda and their sympatric species occurrences by camera trapping and field survey in the Wanglang Nature Reserve, Sichuan, from May 2018 to June 2019. Ten wild species and one domestic species with high photographic presences were selected and clustered by body size indicators. Generalized linear models and redundancy analysis showed that the distribution of giant panda was significantly influenced by the presence of large- and medium-sized species and domestic cattle, but less so by small-sized species. Giant panda tend to avoid cattle by choosing more inferior habitats. Differences in the impact of the ten sympatric species and cattle on the giant panda habitat selection are reflected in three variables: slope, tree density and herb coverage. Our study suggests that; i) more attention should be paid to habitat protection and restoration, particularly for those habitats identified as most suitable for giant pandas; ii) as body size of other species present is a factor which adversely influences giant panda distribution, reductions in large-sized grazing species will be necessary to support giant pandas in nature reserves, and iii) residents should consider replacing cattle with smaller-sized domestic species.

Because of genetic complex and variations of auto antibodies of rheumatoid arthritis (RA) in different populations, the data on the association of HLA-DRB1 alleles in anti-citrullinated protein antibody of (ACPA) RA were inconsistent. The purpose of the study is to systematically summarize results of published data through performing a meta-analysis using data in which HLA-DRB1 alleles are associated with ACPApositive RA and ACPA-negative RA. In this study, we collected data from 12 studies with 13,861 cases and 12,896 controls. Information in these studies included HLADRB1 typing and ACPA status from different countries. Odds ratios (ORs) and corresponding 95% confidence intervals (CI) were used to analyze the association of different HLA-DRB1 alleles with ACPA-positive RA or ACPA-negative RA. To correct skewing data, the analysis of ACPA-status was stratified by patient distribution. Our analyses indicate that in ACPA-positive RA, all patients with RA had significantly higher frequencies of HLA-DRB1*01, *04, *0401, *0405, *07, *11, *13 and *14 than controls. One of the HLA-DRB1*07, *11, *13 and *14 showed protective association with RA. In addition, HLA-DRB1*03, *10 and *12 had more influence than control to RA in European populations; the HLA-DRB1*03 and *12 alleles were associated with the protection. In ACPA-negative RA, only DRB1*07 was associated with the protection (OR 0.53 [95% CI 0.36 - 0.76]) among all HLA-DRB1 alleles in European populations. In ACPA-positive RA, currently available results indicate that *01, *04, *0401 and *0405 are susceptible, while HLA-DRB1*07, *11, *13 and *14 are protective in all populations. While the HLA-DRB1*10 is susceptible, HLA-DRB1*03 and *12 show protective association with RA in European populations. Additionally, regardless of the positive or negative ACPA, the DRB1*07 is always associated with protection in European populations.

Rheumatoid arthritis (RA) and periodontal disease (PD) are chronic inflammatory diseases that share similar osteoclasia, human leukocyte antigen-DR4 allelic genes and immunological profile, and characteristic cytokines. Smoking can contribute to more severe RA and PD; secretion of pro-inflammatory mediators destroys the soft synovial membrane and periodontium, respectively. Anti-citrullinated protein antibodies and anti-α-enolase antibody are characteristic of these two diseases. Some studies suggest that PD may be associated with RA. Anti-Porphyromonas gingivalis (P. gingivalis) antibody, but no P. gingivalis bacterium can be detected in RA patients' joint fluid. Anti-P. gingivalis antibody has been seen as a biomarker of RA. Both diseases share some nosogenesis and common pathological pathways. However, there are differing views on the connection between the two diseases. Interferon-inducible-16 (IFI16) is a genic marker of RA; moreover, the association between IFI16 and PD is rare. Some studies suggest PD is related to periodontal parameters and patient's pathological status rather than RA. Disease frequency in men and women differ between these two diseases. The expression of interleukin-17 (IL-17) receptor only associates with different genders in PD (PD of different sexes have different IL-17 expressions). Periodontal local treatment only affects clinical periodontal status, and it does not alter circulating levels of IL-6, tumor necrosis factor-alpha or C-reactive protein which are associated with RA. This review examines the similarities and differences between these two diseases and explores possible interactions. Importantly, we will discuss whether PD is a feature of RA and whether this knowledge provides helpful information in future treatment of both diseases.

To clarify the effects of gap size and age, as well as microsites on Abies faxoniana recruitment in gaps with or without dwarf bamboo, the occurrences of A. faxoniana seedlings and saplings on four microsites were examined in a subalpine forest in Wanglang Natural Reserve, southwestern China. Results showed that: (1) while increased gap size had little effect on the seedling densities, it significantly reduced the sapling densities on moss-cover ground in A gaps (without bamboos) and those on moss-cover ground and decaying logs in AF gaps (with bamboos; P < 0.05). (2) Increased gap age also exerted little effect on the seedling densities while significantly reduced the sapling densities on decaying logs in AF gaps (P < 0.05). (3) Most of A. faxoniana seedlings and saplings occurred on decaying logs in AF gaps, but in A gaps A. faxoniana regeneration had no preference on microsites. Compared to gap characteristics, microsites are more important for A. faxoniana regeneration.

Ticks are important vectors in the transmission of a broad range of micropathogens to vertebrates, including humans. Because of the role of ticks in disease transmission, identifying and characterizing the micropathogen profiles of tick populations have become increasingly important. The objective of this study was to survey the micropathogens of Hyalomma rufipes ticks. Illumina HiSeq2000 technology was utilized to perform deep sequencing of small RNAs (sRNAs) extracted from field-collected H. rufipes ticks in Gansu Province, China. The resultant sRNA library data revealed that the surveyed tick populations produced reads that were homologous to St. Croix River Virus (SCRV) sequences. We also observed many reads that were homologous to microbial and/or pathogenic isolates, including bacteria, protozoa, and fungi. As part of this analysis, a phylogenetic tree was constructed to display the relationships among the homologous sequences that were identified. The study offered a unique opportunity to gain insight into the micropathogens of H. rufipes ticks. The effective control of arthropod vectors in the future will require knowledge of the micropathogen composition of vectors harboring infectious agents. Understanding the ecological factors that regulate vector propagation in association with the prevalence and persistence of micropathogen lineages is also imperative. These interactions may affect the evolution of micropathogen lineages, especially if the micropathogens rely on the vector or host for dispersal. The sRNA deep-sequencing approach used in this analysis provides an intuitive method to survey micropathogen prevalence in ticks and other vector species.

Vitamin D (VD) and vitamin C (VC) have been recognized as essential elements for human health. Animal models, especially mouse models, have been widely used in the study of VD and VC. This study is to investigate how VD and VC interact at molecular level using whole genome expression profiles from spleens of 111 mouse strains and livers from 40 mouse strains. We first identified the genes that are closely correlated to vitamin D receptor (Vdr) and gulonolactone oxidase (Lgo). We next analyzed the potential molecular pathways of Vdrand Lgo correlated genes and examined how these two sets of pathways are connected in liver and spleen. Our results indicated that Vdrand Lgo associate to distinct groups of genes. While most of genes are not the same, a few of them are associated to both Vdrand Lgo. Our gene network construction suggests that there are interactions between Vdrand Lgopathways. Our work laid down the foundation for future study of the interactions between VD and VC and revealed the potential bias when using animals for the study of VD or VC, as humans do not produce VC.

Colorectal cancer is one of the most common causes of cancer-related death in humans. Coptisine (COP) is a natural alkaloid from Coptidis Rhizoma with unclear antitumor mechanism. Human colon cancer cells (HCT-116) and xenograft mice were used to systematically explore the anti-tumor activity of COP in this study. The results indicated that COP exhibited remarkably cytotoxic activities against the HCT-116 cells by inducing G1-phase cell cycle arrest and increasing apoptosis, and preferentially inhibited the survival pathway and induced the activation of caspase proteases family of HCT-116 cells. Experimental results on male BALB/c nude mice confirmed that orally administration of COP at high-dose (150 mg/kg) could suppress tumor growth, and may reduce cancer metastasis risk by inhibiting the RAS-ERK pathway in vivo. Taken together, the results suggested that COP may be potential as a novel anti-tumor candidate in the HCT-116 cells-related colon cancer, further studies are still needed to suggest COP for the further use.

Actinomycetes, Bacillus subtilis, and Bacillus thuringiensis were isolated from municipal–solid waste (MSW) compost, and different microbial liquid filtrates (MLF) were prepared. Sterile culture media with no microbes were used as their controls. The effects of MLF on soil nematode communities were examined in pot-grown Festuca arundinacea Schreb. Fifteen genera of nematodes in background soil were identified, of which Helicotylenchus and Rotylenchus were dominant. The inoculation of MLF strongly affected the abundance and community structure of soil nematodes. Compared with their controls, lower total nematode numbers following MLF incorporation were found. Actinomycetes inoculation changed community structure of soil nematodes, transforming the dominant genera from Helicotylenchus and Rotylenchus into Cephalobus, Chiloplacus, and Aphelenchus. Actinomycetes incorporation resulted in a significant decrease of plant-parasitic nematodes relative to control pots. Only plant-parasitic and omnivorous-predatory nematodes were found in treatments following B. subtilis inoculation, and Helicotylenchus, Rotylenchus were dominant genera with relative abundance of 76.2% and 14.3%, respectively. Although the dominant genera were still Helicotylenchus and Rotylenchus, B. thuringiensis inoculation led to a marked decrease in populations of plant-parasitic nematodes and an increase in populations of fungivorous and bacterivorous nematodes relative to control. Shannon's diversity index (H′), evenness index (J′), richness index (SR), and Wasilewska index (WI) in pots treated with actinomycetes and B. thuringiensis filtrates were significantly higher than those of their controls, whereas significant lower dominance index (λ) in actinomycetes and B. thuringiensis treatments was observed than their controls. Plant growth was improved in the treatments inoculated with three microbes. The findings highlight that actinomycetes can most effectively suppress plant-parasitic nematodes, increase community diversity, evenness, and richness, thus improving soil environment for turf growth.

Background Plague, a Yersinia pestis infection, is a fatal disease with tremendous transmission capacity. However, the mechanism of how the pathogen stays in a reservoir, circulates and then re-emerges is an enigma. Methodology/Principal findings We studied a plague outbreak caused by the construction of a large reservoir in southwest China followed 16-years’ surveillance. Conclusions/Significance The results show the prevalence of plague within the natural plague focus is closely related to the stability of local ecology. Before and during the decade of construction the reservoir on the Nanpan River, no confirmed plague has ever emerged. With the impoundment of reservoir and destruction of drowned farmland and vegetation, the infected rodent population previously dispersed was concentrated together in a flood-free area and turned a rest focus alive. Human plague broke out after the enzootic plague via the flea bite. With the construction completed and ecology gradually of human residential environment, animal population and type of vegetation settling down to a new balance, the natural plague foci returned to a rest period. With the rodent density decreased as some of them died, the flea density increased as the rodents lived near or in local farm houses where had more domestic animals, and human has a more concentrated population. In contrast, in the Himalayan marmot foci of the Qinghai-Tibet Plateau in the Qilian Mountains. There are few human inhabitants and the local ecology is relatively stable; plague is prevalence, showing no rest period. Thus the plague can be significantly affected by ecological shifts., Author summary Plague in China is shown to be in 12 kinds of natural foci with a large scale geographic and complex structure. Each plague focus has a unique ecological environment, specific geographic regions, landscape characteristics and specific hosts and vectors for the maintenance and transmission of Y. pestis. Once the environment alters, the amount and density of hosts and vectors change and so does the survival of Y. pestis. Consequently, a rest or slightly active foci can com alive, rapidly cause plague among animals and/or humans and then disappear for long periods of time. A plague outbreak among animals and humans caused by the construction of a large reservoir in southwest China is reported in this study. The reservoir was located on the border between Yunnan-Guizhou Plateau and the Guangxi Hills on the Nanpan River, and the reservoir was not previously believed to contain the plague.

Fungal rhinosinusitis (FRS) is commonly caused by various Aspergillus species (spp) and Mucorales fungi, and the treatment and prognosis of cases differ depending on the causative fungus. The present study describes a novel immunohistochemical method that has high sensitivity and specificity for distinguishing between these two types of fungi in patients with FRS. Three groups were included in the study. Group A included formalin‑fixed paraffin‑embedded blocks of 51 nasal tissue specimens of patients with FRS (27 Aspergillus spp and 24 Mucorales) that were continuously obtained from the Department of Pathology of Tongren Hospital in Beijing as the experimental group and 34 cultures (26 Aspergillus spp and 8 Mucorales) of FRS that were randomly selected from the bacterial laboratory of Tongren Hospital in Beijing to verify the staining results of the paraffin‑embedded blocks. Formalin‑fixed paraffin‑embedded blocks of 10 esophageal cancer specimens were included in Group B as the positive control group. All specimens in Groups A and B were stained with interferon‑γ (IFN‑γ) antibody. Group C consisted of the same specimens as described in Group A, however, when performing the immunohistochemical assay, IFN‑γ antibody was replaced by PBS and this served as the negative control group. The differences in IFN‑γ immunohistochemical staining between Aspergillus spp and Mucorales were analyzed. Staining of IFN‑γ in paraffin‑embedded samples was positive in 92.6% (25/27) of specimens in which Aspergillus spp were the causative pathogen, which was significantly higher compared with specimens in which Mucorales was causative (P

Aim of study: to study the effects of gap size, gap age and bamboo Fargesia denudata on natural regeneration of Abies faxoniana, both of which are the ubiquitous dominants in our research area.Area of study: subalpine coniferous forests in Wanglang Natural Reserve in Southwestern China.Material and Methods: 10 transect belts were randomly established, and a total of 97 gaps were recorded and used.Main results: (1) the number of bamboos with coverage of F. denudata with coverage of >17%. (2) F. denudata strongly inhibited A. faxoniana seedlings and saplings in small, young and old gaps, where the amount of A. faxoniana recruitment was relatively abundant, than in other types of gap. (3) The numbers of A. faxoniana seedlings in A-gaps, significantly decreased with the increases in gap size. However, in gaps where F. denudate was also present, A. faxoniana seedlings and saplings were insensitive to gap size or age. Research highlights: thick F. denudata would not be influenced by gap size or age. Because of the low occurrences of A. faxoniana seedlings and saplings, the negative effect of gap size, gap age and F. denudata on A. faxoniana recruitment was unclear.Key words: Abies faxoniana; Fargesia denudata; gap age; gap size; regeneration.

Tightly regulated Ca2+ homeostasis is a prerequisite for proper cardiac function. To dissect the regulatory network of cardiac Ca2+ handling, we performed a chemical suppressor screen on zebrafish tremblor embryos, which suffer from Ca2+ extrusion defects. Efsevin was identified based on its potent activity to restore coordinated contractions in tremblor. We show that efsevin binds to VDAC2, potentiates mitochondrial Ca2+ uptake and accelerates the transfer of Ca2+ from intracellular stores into mitochondria. In cardiomyocytes, efsevin restricts the temporal and spatial boundaries of Ca2+ sparks and thereby inhibits Ca2+ overload-induced erratic Ca2+ waves and irregular contractions. We further show that overexpression of VDAC2 recapitulates the suppressive effect of efsevin on tremblor embryos whereas VDAC2 deficiency attenuates efsevin's rescue effect and that VDAC2 functions synergistically with MCU to suppress cardiac fibrillation in tremblor. Together, these findings demonstrate a critical modulatory role for VDAC2-dependent mitochondrial Ca2+ uptake in the regulation of cardiac rhythmicity. DOI: http://dx.doi.org/10.7554/eLife.04801.001, eLife digest The heart is a large muscle that pumps blood around the body by maintaining a regular rhythm of contraction and relaxation. If the heart loses this regular rhythm it works less efficiently, which can lead to life-threatening conditions. Regular heart rhythms are maintained by changes in the concentration of calcium ions in the cytoplasm of the heart muscle cells. These changes are synchronised so that the heart cells contract in a controlled manner. In each cell, a contraction begins when calcium ions from outside the cell enter the cytoplasm by passing through a channel protein in the membrane that surrounds the cell. This triggers the release of even more calcium ions into the cytoplasm from stores within the cell. For the cells to relax, the calcium ions must then be pumped out of the cytoplasm to lower the calcium ion concentration back to the original level. Shimizu et al. studied a zebrafish mutant—called tremblor—that has irregular heart rhythms because its heart muscle cells are unable to efficiently remove calcium ions from the cytoplasm. Embryos of the tremblor mutant were treated with a wide variety of chemical compounds with the aim of finding some that could correct the heart defect. A compound called efsevin restores regular heart rhythms in tremblor mutants. Efsevin binds to a pump protein called VDAC2, which is found in compartments called mitochondria within the cell. Although mitochondria are best known for their role in supplying energy for the cell, they also act as internal stores for calcium. By binding to VDAC2, efsevin increases the rate at which calcium ions are pumped from the cytoplasm into the mitochondria. This restores rhythmic calcium ion cycling in the cytoplasm and enables the heart muscle cells to develop regular rhythms of contraction and relaxation. Increasing the levels of VDAC2 or another similar calcium ion pump protein in the heart cells can also restore a regular heart rhythm. Efsevin can also correct irregular heart rhythms in human and mouse heart muscle cells, therefore the new role for mitochondria in controlling heart rhythms found by Shimizu et al. appears to be shared in other animals. The experiments have also identified the VDAC family of proteins as potential new targets for drug therapies to treat people with irregular heart rhythms. DOI: http://dx.doi.org/10.7554/eLife.04801.002

An outbreak of Streptococcus suis serotype 2 emerged in the summer of 2005 in Sichuan Province, and sporadic infections occurred in 4 additional provinces of China. In total, 99 S. suis strains were isolated and analyzed in this study: 88 isolates from human patients and 11 from diseased pigs. We defined 98 of 99 isolates as pulse type I by using pulsed-field gel electrophoresis analysis of SmaI-digested chromosomal DNA. Furthermore, multilocus sequence typing classified 97 of 98 members of the pulse type I in the same sequence type (ST), ST-7. Isolates of ST-7 were more toxic to peripheral blood mononuclear cells than ST-1 strains. S. suis ST-7, the causative agent, was a single-locus variant of ST-1 with increased virulence. These findings strongly suggest that ST-7 is an emerging, highly virulent S. suis clone that caused the largest S. suis outbreak ever described.

RNA nanotechnology promises rational design of RNA nanostructures with wide array of structural diversities and functionalities. Such nanostructures could be used in applications such as small interfering RNA delivery and organization of in vivo chemical reactions. Though having impressive development in recent years, RNA nanotechnology is still quite limited and its programmability and complexity could not rival the degree of its closely related cousin: DNA nanotechnology. Novel strategies are needed for programmed RNA self-assembly. Here, we have assembled RNA nanocages by re-engineering a natural, biological RNA motif: the packaging RNA of phi29 bacteriophage. The resulting RNA nanostructures have been thoroughly characterized by gel electrophoresis, cryogenic electron microscopy imaging and dynamic light scattering.

The present study aimed at investigating the structural features and antioxidant activities of a polysaccharide fraction (DHP1A) obtained from Dendrobium huoshanense, a precious herb medicine in China. DHP1A mainly consisted of mannose (Man), glucose (Glc) and a trace of galactose (Gal), with a molecular weight of 6700Da. Its backbone contained (1→4)-linked α-D-Glcp, (1→6)-linked α-D-Glcp and (1→4)-linked β-D-Manp, with a branch of terminal β-D-Galp. The in vitro antioxidant evaluation revealed that DHP1A had a remarkable inhibition effect on the FeCl2-induced lipid peroxidation. Furthermore, DHP1A pretreatment decreased the production of malondialdehyde (MDA), and restored the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), as well as the level of glutathione (GSH) in the livers of CCl4-treated mice. These results suggested that DHP1A was a potential antioxidant component in D. huoshanense.

Premise of the study: There is a need for microsatellite primers to analyze genetic parameters of Fargesia denudata (Poaceae), the staple-food bamboo of the giant panda (Ailuropoda melanoleuca). Methods and Results: Using next-generation sequencing technology, we obtained a 75-Mb assembled sequence of F. denudata and identified 182 microsatellites. Primer pairs for 70 candidate microsatellite markers were selected and validated in four individuals, and 42 primer pairs generated reliable amplicons. Fourteen of 16 tested markers were found to be polymorphic in 72 individuals from four F. denudata populations. The number of alleles ranged from two to 19 per locus; the observed and expected heterozygosities ranged from 0 to 1 and from 0 to 0.87, respectively. The transferability of these 16 novel microsatellite markers was validated in five related species. Conclusions: These markers will be useful for examining the genetic diversity, genetic structure, and cloning of F. denudata, the staple-food bamboo of th...

Introduction Successful root canal treatment is dependent on detailed knowledge of the morphology of the root canal system. The aim of this study was to investigate the morphology of the root canal systems of mandibular first premolars and the location of root canal orifices and apical foramina in a Chinese subpopulation using cone-beam computed tomography scanning. Methods A total of 440 cone-beam computed tomographic images of mandibular first premolars were obtained from 238 patients. Tooth position, root canal morphology, the distance between the apical foramen and the anatomic apex, and the distance between the 2 root canal orifices of mandibular first premolars with 2 root canals were investigated. The Fisher exact test was used to analyze the correlation between the number of root canals and tooth position. Results Overall, 1.14% ( n = 5) of the teeth had a C-shaped root canal system, 77.14% ( n = 335) had 1 root canal, and 22.05% ( n = 97) had 2 root canals from type II to type V. The remaining 0.68% ( n = 3) were classified as type VIII, which had 3 root canals. The most frequent distribution of the distance between the apical foramen and the anatomic apex in 335 mandibular first premolars with 1 root canal was 1–2 mm ( n = 192). The distance between the apical foramen and the anatomic apex of mandibular first premolars with 2 root canals was in accord with that of mandibular first premolars with 1 root canal. The distance between the 2 root canal orifices of mandibular first premolars with 2 root canals was approximately 13 mm. Conclusions Most mandibular first premolars had 1 root canal. More than 1 of 5 mandibular first premolars had complex root canal systems (with 2 or more root canals). This study provides supplemental information about the root canals of mandibular first premolars in a Chinese subpopulation.

OBJECTIVE To distinguish the ginseng and American ginseng pieces accurately and rapidly by electronic nose technology and principal component analysis (PCA) method. METHOD The optimum conditions of electronic nose for ginseng and American ginseng pieces, such as sample size and volume, headspace volume, incubation time and temperature were determined by the orthogonal test, the data were processed by the normalization method and the preprocessed data were analyzed PCA. RESULT The detection methods of ginseng and American ginseng pieces was established by electronic nose, and the odor fingerprint figures of ginseng and American ginseng pieces were obtained, and ginseng and American ginseng pieces were distinguished by PCA recognition pattern. CONCLUSION A new accurate and rapid method to distinguish ginseng and American ginseng pieces was established by electronic nose detection.

Background Gain of 3q26 is frequently observed in squamous cell carcinomas of mucosal origin, including those originating in the head and neck region. The human telomerase RNA component (hTERC) gene, which is located on chromosome 3q26, encodes for an RNA subunit of telomerase that maintains the length of telomeres through cellular divisions, and is activated in malignant diseases. The present study was designed to detect hTERC amplification in laryngeal lesions and evaluate whether this might serve as a supportive biomarker in histopathological analysis for in the diagnosis of laryngeal lesions. Methods Fluorescent in situ hybridization (FISH) was applied on formalin-fixed paraffin-embedded blocks of 93 laryngeal specimens, including 14 normal epithelium (NE), 15 mild dysplasia (Md), 18 moderate dysplasia (MD), 16 severe dysplasia (SD), 9 carcinoma in situ (CIS), and 21 invasive carcinoma (IC)). Results By histopathologic examination, hTERC amplification rates in NE, Md, MD, SD, CIS and IC cases were 0% (0/14), 13.33% (2/15), 72.22% (13/18), 81.25% (13/16), 100% (9/9) and 100% (21/21), respectively. Amplification of hTERC was significantly associated with histopathologic diagnosis (P < 0.0001). The percentage of hTERC amplification in patients with MD, SD, CIS, and IC was significantly higher than those with NE or Md (P < 0.0001). The number of cells with abnormal signals increased and the abnormal signal patterns were diversified with increasing severity of laryngeal dysplasia (P < 0.0001). Conclusions The hTERC amplification is important in the development of laryngeal squamous cell carcinoma (LSCC). FISH detection of hTERC amplification may provide an effective approach in conjunction with histopathologic evaluation for differential diagnosis of laryngeal lesions. Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2226606266791985

Introduction Human dental apical papilla cells (APCs) have mineralization potential, which plays a key role in the root development of young permanent teeth. Limited literature is available about APC mineralization in the presence of inflammatory cytokines. The purpose of this study was to investigate the effects of tumor necrosis factor-α (TNF-α) on APC mineralization. Methods APC cultures were established with the enzymatic dissociation method in vitro . The viability of APCs treated with TNF-α was investigated using methyl-thiazol-tetrazolium assays. Cells were then cultured in osteo-/dentinogenic medium with TNF-α, and mineralization was assessed by alizarin red S staining. Bone sialoprotein (BSP) and dentin sialoprotein (DSP) were analyzed using immunocytochemistry. Mineralization genes such as BSP, dentin sialophosphoprotein (DSPP), osteocalcin (OCN), and dentin matrix acidicphosphoprotein-1 (DMP1) were determined with real-time polymerase chain reaction analyses. Results The viability of cultured cells was higher with TNF-α concentrations of 10 ng/mL and 50 ng/mL than with 5 ng/mL or in the control group. Alizarin red S staining showed that APCs had a higher mineralization activity when the osteo-/dentinogenic culture medium contained 10 ng/mL TNF-α. Immunocytochemical detection showed that the expression of BSP and DSP was positive in APCs after they were induced in osteo-/dentinogenic medium. The expression of mineralization genes differed when treated with 10 ng/mL TNF-α (ie, the expression of DSPP mRNA increased on days 7 and 14, whereas the expression of DSPP mRNA decreased on day 21). Conclusions TNF-α may promote APC mineralization in short-term cultures and inhibit the mineralization in long-term cultures.

Most studies concerning the initiation and development of the shoot apical meristem have been performed on meristems that developed during embryogenesis. We characterized the in situ formation of adventitious shoots originating from cells located near leaf axils in a series of six interspecific periclinal tobacco chimeras. Shoots were generated by decapitating the plants and removing all of the axillary buds. Eighty-four of the 413 shoots regenerated were chimeral. Many of the shoots were complex mericlinal chimeras, with several of their axillary buds possessing meristerns arranged as periclinal chimeras. The adventitious shoots originated from derivatives of only the second and third cell layers in the meristem of the source plant since derivatives of the outermost meristem layer were removed with the removal of the axillary buds. With time, nearly all adventitious shoot apices that were initially chimeral became nonchimeral or stabilized as periclinal chimeras. A statistical analysis indicated that the arrangement of genetically distinct tissues in the cell layers of adventitious meristems was influenced by competition between cell types. Our method for generating these shoots can be used to create small genetically distinct sectors analogous to radiation-induced sectors as well as a complete series of periclinal chimeras, both of which have potential for use in determining tissue-to-tissue interactions. An analysis of sector length and position on chimeral shoots indicates that the first one to three leaves of adventitious shoots do not arise as derivatives of the shoot apical initials of a meristem proper.