Abstract To study the purification conditions of polysaccharide from Solanum nigrum L. by S-8 macroporous adsorption resin. The effect of multi-factors on the purification of polysaccharide from Solanum nigrum were studied by static - dynamic adsorption and desorption methods with indexes of adsorption and desorption rates. The optimized purification process condition was as follows: the concentration of sample was 5 mg/mL, pH level was 7.0, temperature was 30 °C, the adsorption velocity was 0.5 mL/min, the maximun loading amount was 10 BV, 0.5 mol/L NaCl solution 5BV was used as eluent, elution flow rate was 4 mL/min. With this condition, the purity of Solanum nigrum polysaccharide was 91.00%, which was 1.989 times higher than before. S-8 macroporous resin can be used to separate and purify the solanum nigrum polysaccharide and improve the purity and quality of polysaccharides.

Abstract Lactic acid bacteria (LAB) can produce extracellular polysaccharides that can be used as thickeners, emulsifiers, and gels in foods. The immobilization technology can effectively simplify and improve the separation and regeneration of extracellular polysaccharides. In this study, six strains of LAB, including the extracellular polysaccharide-producing strain Lactobacillus plantarum, were screened. The different immobilized carriers of sodium alginate, agar, gelatin-glutaraldehyde, and carrageenan were assessed for fermentation by these six LAB. Different diameters of beads were selected, and the optimum culture temperature, carrier concentration, and inoculum volume were evaluated. The results showed that the agar-embedding method was the best immobilization method and the optimum process conditions were as follows: 3% inoculum of L. plantarum, 30 g/L of agar as immobilization carrier with a particle diameter of 1 cm, and fermentation at 37 °C for 24 h. Through orthogonal experiments, the maximum exopolysaccharide (EPS) yield obtained was 1489.9 mg/L.

Abstract Background: The internal NAD(P)H dehydrogenase (NDA) gene family was a member of the NAD(P)H dehydrogenase (ND) gene family, mainly involved in the non-phosphorylated respiratory pathways in mitochondria and played crucial roles in response to abiotic stress. Methods: The whole genome identification, structure analysis and expression pattern of NDA gene family were conducted to analyze the NDA gene family. Results: There were 51, 52, 26, and 24 NDA genes identified in G. hirsutum, G. barbadense, G. arboreum and G. raimondii, respectively. According to the structural characteristics of genes and traits of phylogenetic tree, we divided the NDA gene family into 8 clades. Gene structure analysis showed that the NDA gene family was relatively conservative. The four Gossypium species had good collinearity, and segmental duplication played an important role in the evolution of the NDA gene family. Analysis of cis-elements showed that most GhNDA genes contained cis-elements related to light response and plant hormones (ABA, MeJA and GA). The analysis of the expression patterns of GhNDA genes under different alkaline stress showed that GhNDA genes were actively involved in the response to alkaline stress, possibly through different molecular mechanisms. By analyzing the existing RNA-Seq data after alkaline stress, it was found that an NDA family gene GhNDA32 was expressed, and then theGhNDA32 was silenced by virus-induced gene silencing (VIGS). By observing the phenotype, we found that the wilting degree of silenced plants was much higher than that of the control plant after alkaline treatment, suggesting that GhNDA32 gene was involved in the response to alkaline stress. Conclusions: In this study, GhNDAs participated in response to alkaline stress, especially NaHCO3 stress. It was of great significance for the future research on the molecular mechanism of NDA gene family in responding to abiotic stresses.

This study describes a virtual screening performed for two series of selenides (28 compounds), derived from N-phenylacetamides chlorides and 7-chloro-quinoline, to determine their potential for leishmanicidal activity against Leishmania amazonensis and Leishmania donovani. Seven compounds were predicted as potential leishmanicides; therefore, they were synthesized from elemental selenium, as a precursor for the production of NaHSe, and subsequent reactions with 4,7-dichloro-quinoline and N-phenylacetamides chlorides were performed. The compounds were characterized by infrared (IR), 1H and 13C nuclear magnetic resonance (NMR), and sent for in vitro cytotoxicity tests against L. amazonensis and were found to be active and selective, and two compounds presented half-maximal inhibitory concentrations (IC50) of 5.67 and 10.81 µg mL-1. They also presented good interaction energies in the docking study, suggesting that may exert their effects by inhibiting the N-myristoyltransferase and O-acetylserine sulfhydrylase enzymes in parasites.

ABSTRACT The response mechanism and interaction patterns of HIF-1α and p53 in animals in an hypoxic environment are crucial for their hypoxic tolerance and adaptation. Many studies have shown that underground rodents have better hypoxic adaptation characteristics. However, the mechanism by which HIF-1α and p53 in underground rodents respond to hypoxic environments compared with in ground rodents remains unclear. Further, whether a synergy between HIF-1α and p53 enables animals tolerate extremely hypoxic environments is unclear. We studied HIF-1α and p53 expression in the brain tissue and cell apoptosis in the hippocampal CA1 region during 6 hours of acute hypoxia (5% oxygen) in Lasiopodomys mandarinus (Milne-Edwards, 1871) and Lasiopodomys brandtii (Radde, 1861), two closely related small rodents with different life characteristics (underground and aboveground, respectively), using a comparative biology method to determine the mechanisms underlying their adaptation to this environment. Our results indicate that HIF-1α and p53 expression is more rapid in L. mandarinus than in L. brandtii under acute hypoxic environments, resulting in a significant synergistic effect in L. mandarinus. Correlation analysis revealed that HIF-1α expression and the apoptotic index of the hippocampal CA1 regions of the brain tissues of L. mandarinus and L. brandtii, both under hypoxia, were significantly negatively and positively correlated, respectively. Long-term existence in underground burrow systems could enable better adaptation to hypoxia in L. mandarinus than in L. brandtii. We speculate that L. mandarinus can quickly eliminate resulting damage via the synergistic effect of p53 and HIF-1α in response to acute hypoxic environments, helping the organism quickly return to a normal state after the stress.

Abstract Background: Liriodendron chinense ranges widely in subtropical China and northern Vietnam; however, it inhabits several small, isolated populations and is now an endangered species due to its limited seed production. The objective of this study was to develop a set of nuclear SSR (simple sequence repeats) and multiple chloroplast genome markers for genetic studies in L. chinense and their characterization in diverse germplasm. Results: We performed low-coverage whole genome sequencing of the L. chinense from four genotypes, assembled the chloroplast genome and identified nuclear SSR loci by searching in contigs for SSR motifs. Comparative analysis of the four chloroplast genomes of L. chinense revealed 45 SNPs, 17 indels, 49 polymorphic SSR loci, and five small inversions. Most chloroplast intraspecific polymorphisms were located in the interspaces of single-copy regions. In total, 6147 SSR markers were isolated from low-coverage whole genome sequences. The most common SSR motifs were dinucleotide (70.09%), followed by trinucleotide motifs (23.10%). The motif AG/TC (33.51%) was the most abundant, followed by TC/AG (25.53%). A set of 13 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 109 L. chinense individuals, representing distinct varieties or germplasm. The number of alleles per locus ranged from 8 to 28 with an average of 21 alleles. The expected heterozygosity (He) varied from 0.19 to 0.93 and the observed heterozygosity (Ho) ranged from 0.11 to 0.79. Conclusions: The genetic resources characterized and tested in this study provide a valuable tool to detect polymorphisms in L. chinense for future genetic studies and breeding programs.

Abstract: Background: The extension of the blueberry industry in China is restricted by the low performance of proliferation coefficient, transplanting survival rate and the long cycle production. Hypothesis: We hypothesized the suitable medium with optimal concentration and type combination would improve the survival rate of Vaccinium corymbosum. Species studied: Vaccinium corymbosum L. cultivar ‘O’Neal. Study site and years of study: Our study was conducted in Yunnan Breeding and Cultivation Research and Development Center of Endangered and Daodi Chinese medicinal materials, Yunnan University of Chinese Medicine since November 2015. Methods: Efficient protocol of plant regeneration about Southern highbush blueberry (cultivar ‘O’Neal) was established using annual shoots via single factor and orthogonal experiments. Results: Olive medium with 2.0 mg·L-1 zeatin was found to be most suitable for initiation culture. The highest callus induction and bud formation rate were determined with 93.67 % and 95.57 %, respectively. Furthermore, as the germination of axillary buds, numerous shoots were formed with the proliferation coefficient (> 60). Well-developed shoots were obtained using rejuvenation culture on half-strength Olive medium supplemented with combination of 2.0 mg·L-1 indole-3-butyric acid, 1.0 mg·L-1 naphthaleneacetic acid, 0.01 mg·L-1 chlormequat chloride and 0.5 g·L-1 activated charcoal. The rooting rate could reach to 100 % using 2 cm length of well-developed shoots transferred into the 5:1 sterilized peat:perlite, which was enhanced from 27.61 up to 95 % after transplanting to the field. Conclusions: An efficient system for plant regeneration from bud induction to transplanting to the field was established to maintain the characteristics of southern highbush blueberry varieties.

Resumen: Antecedentes: La ampliación de la industria del arándano en China está restringida por bajo rendimiento del coeficiente de proliferación, tasa de supervivencia del trasplante y ciclos de producción largo. Hipótesis: Un medio adecuado adicionado con tipos y concentración óptima de auxina mejorará la tasa de supervivencia de Vaccinium corymbosum. Especies estudiadas: Vaccinium corymbosum L. cultivar ’O’Neal. Sitio y años de estudio: Centro de Investigación y Desarrollo de Reproducción y Cultivo de Materiales Medicinales Chinos en Peligro de Extinción y “Daodi” de Yunnan, Universidad de Medicina China de Yunnan, desde noviembre de 2015. Métodos: Se estableció un protocolo eficiente de regeneración de plantas de arándano azul del sur (cultivar 'O’Neal) utilizando brotes anuales vía factor único y experimentos ortogonales. Resultados: El medio de oliva con 2.0 mg de L-1 de zeatina fue más adecuado para iniciar el cultivo. Aumentó en 93.67 y 95.57 % la inducción de callo y tasa de formación de brotes, respectivamente. La germinación de los brotes axilares provocó numerosos brotes con un coeficiente de proliferación > 60. Se obtuvo brotes bien desarrollados usando cultivos de rejuvenecimiento con medio de oliva adicionado con 2.0 mg·L-1 de ácido indol-3-butírico, 1.0 mg·L-1 de ácido naftalenacético, 0.01 mg L-1 de cloruro de clormequato L-1 y 0.5 g L-1 de carbón activado. La tasa de enraizamiento alcanzó 100 % utilizando brotes bien desarrollados de 2 cm de longitud y transferidos a turba-perlita 5:1, aumentando de 27.61 a 95 % después del trasplante. Conclusiones: Se estableció un sistema eficiente de regeneración a partir de inducción de yemas hasta su establecimiento en el campo, para mantener las características de las variedades de arándanos del sur.

ABSTRACT Grain shape in rice is a key determinant of grain appearance, yield and market value, and thus has been widely studied. Rice mutant lines with long-grain phenotypes were previously isolated from an M2 population derived from mutagenized mature pollen grains that were treated with gamma-ray irradiation for a cultivar Ma85. To understand the genetic basis underlying the long-grain trait, two mutant lines, JF171 and JF178, were crossed with the short-grain parents JF222 and Samba, generating three F2 populations. Molecular marker-based genetic analysis was employed to detect the major QTLs that affected grain length, grain width, length-to-width ratio and grain weight. Based on the data obtained for the three populations, a joint major QTL for grain length was mapped to a 4.7 Mb region between the SSR makers RM263 and RM318 on the long arm of chromosome 2. The results suggested that the two independent mutations contained in JF171 and JF178 are likely due to alterations in the same genetic region. Furthermore, we developed a BC2F2 population using JF178 and Samba and narrowed the region to 0.6 Mb. The results of the current study will be helpful to reveal the genetic basis of the above long-grain mutant lines. In general, our results provide solid genetic data to identify the unknown gene that affects grain length by map-based cloning and practice marker-assisted breeding for long-grain rice cultivars.

The majority of goats in Tanzania belong to the Small East African (SEA) breed, which exhibits large phenotypic variation. This study aimed to determine the genetic structure of, and relationships among four populations (Sukuma, Gogo, Sonjo, and Pare) of the SEA breed that have not been studied adequately. A total of 120 individuals (24 from each population) were analysed at eight microsatellite loci. In addition, 24 goats of the South African Boer breed were used as reference. Observed heterozygosity (Ho) ranged from 0.583 ± 0.04 for Sukuma to 0.659 ± 0.030 for Gogo, while expected heterozygosity (He) ranged from 0.632 ± 0.16 for Sukuma to 0.716 ± 0.16 for Boer. Five loci deviated from Hardy-Weinberg equilibrium (HWE) across populations. The mean number of alleles ranged from 4.75 ± 1.58 for Pare to 6.88 ± 3.00 for Sukuma. The mean inbreeding coefficient (F IS) ranged from 0.003 in Sonjo to 0.148 in Sukuma. The differentiation coefficient (F ST)was highest (0.085) between Boer and Sukuma and lowest (0.008) between Gogo and Sonjo. The largest genetic distance (0.456) was found between Sukuma and Boer, while the smallest (0.031) was between Gogo and Sonjo populations. Pare, Gogo, and Sonjo populations, formed one cluster, while Sukuma and Boer populations formed two separate clusters. From the findings, it can be concluded that the SEA goats in this study showed high in population genetic variation, which implies that there is good scope for their further improvement through selection within populations. The Sukuma population, which has fairly high inbreeding, is moderately differentiated from Pare, Sonjo, and Gogo goat populations, which showed a high level of admixture. Conservation and improvement strategies of the goats should be designed with first priority being on Sukuma goats.

The aim of this study was to evaluate the clinical and radiographic outcomes of upper thoracic (UT) versus lower thoracic (LT) upper instrumented vertebrae (UIV) for adult scoliosis by meta-analysis. We conducted a literature search in three databases to retrieve related studies up to March 15, 2017. The preliminary screened studies were assessed by two reviewers according to the selection criteria. All analyses were carried out using the statistical software package R version 2.31. Odds ratios (OR) with 95% confidence intervals (CI) were used to describe the results. The I2 statistic and Q statistic test were used for heterogeneity assessment. Egger's test was performed to detect publication bias. To assess the effect of each study on the overall pooled OR or standardized mean difference (SMD), sensitive analysis was conducted. Ten trials published between 2007 and 2015 were eligible and included in our study. Meta-analysis revealed that the UT group was associated with more blood loss (SMD=0.4779, 95%CI=0.3349-0.6209, Z=6.55, P<0.0001) and longer operating time (SMD=0.5780, 95%CI=0.1971-0.958, Z=2.97, P=0.0029) than the LT group. However, there was no significant difference in Oswestry Disability Index, Scoliosis Research Society (SRS) function subscores, radiographic outcomes including sagittal vertical axis, lumbar lordosis, and thoracic kyphosis, length of hospital stay, and revision rates between the two groups. No evidence of publication bias was found between the two groups. Fusion from the lower thoracic spine (below T10) has as advantages a shorter operation time and less blood loss than upper thoracic spine (above T10) in posterior long-segment fixation for degenerative lumbar scoliosis.

ABSTRACT Some outer floral organs are unique in gramineous plants, like the sterile lemma and rudimentary glume in rice. However, their development mechanisms are still poorly understood. In this study, we used 4 mutants with long sterile lemma (LSL), named JF11, JF12, JF13 and JNY-7, to be crossed with Aijiaonante (AJNT) and Nipponbare (NIP), respectively. Genetic analysis indicated that LSL trait exhibited recessive heredity and was controlled by a common allele named sl-1(t). Using the method of bulk segregant analysis and linkage analysis between SSR markers and LSL trait based on F2 populations, the sl-1(t) gene was located at the interval between RM20903 and RM20948 on chromosome 7. The interval harbors a known G1 gene, which regulates the sterile lemma trait. The findings of allelic sequencing showed an 11-base deletion in gene G1 happened in the mutants of JF11, JF12 and JF13, which led to a frame-shifting mutation, whereas the mutant of JNY-7 had a base substitution that caused the change of the amino acid residue. Eight substitutions in the ORF and 10 in the upstream region from −1 to −824 were found between Indica and Japonica rice by DNA sequence analysis, but those polymorphisms have no effect on the gene function. In conclusion, we fine mapped the LSL gene, sl-1(t), and found 2 kinds of mutant alleles conferring the gene function and the DNA polymorphisms of G1 between Indica and Japonica rice.

BACKGROUND: Laboratory-enhanced surveillance is critical for rapidly detecting the potential re-emergence of Ebola virus disease. Rapid diagnostic tests (RDT) for Ebola antigens could expand diagnostic capacity for Ebola virus disease. OBJECTIVES: The Guinean National Coordination for Ebola Response conducted a pilot implementation to determine the feasibility of broad screening of patients and corpses with the OraQuick® Ebola RDT. METHODS: The implementation team developed protocols and trained healthcare workers to screen patients and corpses in Forécariah prefecture, Guinea, from 15 October to 30 November 2015. Data collected included number of consultations, number of fevers reported or measured, number of tests performed for patients or corpses and results of confirmatory RT-PCR testing. Data on malaria RDT results were collected for comparison. Feedback from Ebola RDT users was collected informally during supervision visits and forums. RESULTS: There were 3738 consultations at the 15 selected healthcare facilities; 74.6% of consultations were for febrile illness. Among 2787 eligible febrile patients, 2633 were tested for malaria and 1628 OraQuick® Ebola RDTs were performed. A total of 322 OraQuick® Ebola RDTs were conducted on corpses. All Ebola tests on eligible patients were negative. CONCLUSIONS: Access to Ebola testing was expanded by the implementation of RDTs in an emergency situation. Feedback from Ebola RDT users and lessons learned will contribute to improving quality for RDT expansion.

Myocardial ischemia is a major cause of death and remains a disease with extremely deficient clinical therapies and a major problem worldwide. Cold inducible RNA-binding protein (CIRBP) is reported to be involved in multiple pathological processes, including myocardial ischemia. However, the molecular mechanisms of myocardial ischemia remain elusive. Here, we first overexpressed CIRBP by transfection of pc-CIRBP (pcDNA3.1 containing coding sequenced for CIRBP) and silenced CIRBP by transfection of small interfering RNA targeting CIRBP (siCIRBP). pcDNA3.1 and the negative control of siCIRBP (siNC) were transfected into H9C2 cells to act as controls. We then constructed a cell model of myocardial ischemia through culturing cells in serum-free medium with hypoxia in H9C2 cells. Subsequently, AlamarBlue assay, flow cytometry and western blot analysis were used, respectively, to assess cell viability, reactive oxygen species (ROS) level and apoptosis, and expression levels of IκBα, p65 and Bcl-3. We demonstrated that CIRBP overexpression promoted cell proliferation (P<0.001), inhibited cell apoptosis (P<0.05), reduced ROS level (P<0.001), down-regulated phosphorylated levels of IκBα and p65 (P<0.01 or P<0.001), and up-regulated expression of Bcl-3 (P<0.001) in H9C2 cells with myocardial ischemia. The influence of CIRBP knockdown yielded opposite results. Our study revealed that CIRBP could protect H9C2 cells against myocardial ischemia through inhibition of NF-κB pathway.

Fluoride, which is often added to toothpaste or mouthwash in order to protect teeth from decay, may be a novel therapeutic approach for acceleration of periodontal regeneration. Therefore, we investigated the effects of fluoride on proliferation and mineralization in human periodontal ligament cells in vitro. The periodontal ligament cells were stimulated with various concentrations of NaF added into osteogenic inductive medium. Immunohistochemistry of cell identification, cell proliferation, alkaline phosphatase (ALP) activity assay, Alizarin red S staining and quantitative real-time-polymerase chain reaction (RT-PCR) were performed. Moderate concentrations of NaF (50-500 μmol/L) had pro-proliferation effects, while 500 μmol/L had the best effects. ALP activity and calcium content were significantly enhanced by 10 μmol/L NaF with osteogenic inductive medium. Quantitative RT-PCR data varied in genes as a result of different NaF concentrations and treatment periods. We conclude that moderate concentrations of NaF can stimulate proliferation and mineralization in periodontal ligament cells. These in vitro findings may provide a novel therapeutic approach for acceleration of periodontal regeneration by addition of suitable concentrations of NaF into the medication for periodontitis treatment, i.e., into periodontal packs and tissue patches.

Glucagon-like peptide 1 (GLP-1), a kind of gut hormone, is used in the treatment of type 2 diabetes (T2D). Emerging evidence indicates that GLP-1 has anti-inflammatory activity. Chronic inflammation in the adipose tissue of obese individuals is a cause of insulin resistance and T2D. We hypothesized that GLP-1 analogue therapy in patients with T2D could suppress the inflammatory response of macrophages, and therefore inhibit insulin resistance. Our results showed that GLP-1 agonist (exendin-4) not only attenuated macrophage infiltration, but also inhibited the macrophage secretion of inflammatory cytokines including TNF-β, IL-6, and IL-1β. Furthermore, we observed that lipopolysaccharide (LPS)-induced macrophage conditioned media could impair insulin-stimulated glucose uptake. This effect was compensated by treatment with the conditioned media from macrophages treated with the combination of LPS and exendin-4. It was also observed that exendin-4 directly inhibited the activation of NF-κB in macrophages. In conclusion, our results indicated that GLP-1 improved inflammatory macrophage-derived insulin resistance by inhibiting NF-κB pathway and secretion of inflammatory cytokines in macrophages. Furthermore, our observations suggested that the anti-inflammatory effect of GLP-1 on macrophages can contribute to GLP-1 analogue therapy of T2D.