Abstract The effects of α-linolenic acid (ALA) on the proliferation and adhesion of probiotics would be investigated in the present study. Effects of ALA on intestinal flora were studied by animal fecal anaerobic fermentation system in vitro, which were analyzed by high-throughput sequencing. Results showed that treatment with ALA could promote the proliferation of probiotics Lactobacillus and Bifidobacterium, and inhibit the growth of harmful bacteria Enterococcus and E. coli. ALA restored the abnormal intestinal flora caused by high-fat diet, which was beneficial to the improvement of intestinal flora structure. In addition, adhesive characteristics of probiotics to epithelial colon cells NCM460 were detected by plate counting and Gram staining, which indicated that ALA promoted adhesion of probiotics to colonic cells. In conclusion, ALA could promote the proliferation and adhesion of intestinal probiotics, which provides a basis for ALA to exert the healthy activities of intestinal probiotics.

Abstract The authentication of fresh apple juice (FAJ) is a significant issue. However, the main characteristics of FAJ, diluted apple juice (DAJ) with added concentrated apple juice (DAJ+CAJ), and DAJ with added sucrose (DAJ+s) are not different when using classical chemical analyses in juice authentication. In this study, isotope ratio mass spectrometry (IRMS) was used to determine the δD, δ18O, and δ13C in adulterated apple juice for juice authentication. The δD and δ18O values were decreased in adulterated juice. A linear correlation was found between δD and δ18O in apple juices with different amount of adulteration. Therefore, this linear relationship can be used to determine the amount of adulteration that took place. The δ13C value and the content of each component in real apple juice are within specific ranges. The adulteration can be recognized once the δ13C value and the content of each component in the apple juice are out of these ranges. The implementation of this method would help maintain the juice market’s stability and protect consumers’ interests.

Abstract The aim of this study was to determine the resistance mechanism of Pseudomonas aeruginosa to cefoperazone sodium/sulbactam sodium. We retrospectively analyzed the drug resistance of P.a isolated at the First Affiliated Hospital of Guangxi Medical University. Drug-resistant P.a strains were constructed, then wild-type (WT) and drug-resistant (DR) strains were compared using protein and gene microarrays to determine differences between DR and WT strains. The resistance rates of P. aeruginosa during 2013, 2014 and 2015 were 21.2%, 21.4%, and 24.6% respectively. Among 242 protein peaks of WT and DR bacteriophage proteins, 41 were differentially expressed between the two groups. The expression of 26 and 15 proteins were respectively upregulated and downregulated in the DR compared with the WT group. Gene microarray results revealed 679 mutant loci in the DR group, of which 42 with the top 50 Q values were found in the NCBI database. The rate of P.a resistance to cefoperazone sodium/sulbactam sodium remained high between 2013 and 2015. The numbers of different proteins and genetic variations in the DR strains suggested that the resistance mechanism of P.a to cefoperazone sodium/sulbactam sodium involves multiple genes and proteins that might be key to controlling P.a resistance to cefoperazone sodium/sulbactam sodium.

Abstract Ligustrazine is widely used for the treatment of cardiovascular diseases in traditional Chinese medication. It has been reported that Ligustrazine decreases the concentration of intracellular calcium ions (Ca2+); however, the underlying mechanism remains unknown. In the present study, the effect of Ligustrazine on adenosine diphosphate (ADP)-induced platelet aggregation was evaluated using a turbidimetric approach. The changes in concentration of intracellular Ca2+ stimulated by ADP was measured using fluo-4, a fluorescent Ca2+ indicator dye. The mRNA expression of stromal interaction molecule l (STIM1) and Orai1, calcium sensor, was determined using real-time PCR. In addition, the protein expression of STIM1, Orai1, and serum/glucocorticoid-regulated protein kinase 1 (SGK1) was determined using Western blot analysis. The data demonstrated that Ligustrazine significantly suppressed platelet aggregation in a dose-dependent manner and reduced the concentration of intracellular Ca2+ triggered by ADP. Our data showed that Ligustrazine treatment inhibited the expression of STIM1 and Orai1 induced by ADP at both mRNA and protein levels, and suppressed the protein expression of SGK1. Taken together, our data indicated that Ligustrazine suppressed platelet aggregation by partly inhibiting the activities of calcium sensors, thereby suggesting that Ligustrazine may be a promising candidate for the treatment of platelet aggregation.

Abstract The effects of 6-gingerol on adhesive ability of probiotics to colonic epithelial cells in vitro were explored. Two strains of probiotics were Lactophilus acidophilus and Bifidobacterium bifidum. NCM 460 cells (normal colonic epithelial cells) and Caco-2 cells (colon cancer cells) were used in this study. Effect of 6-gingerol (5, 10, 15, 20, 25, 30, 35, 40 μg/mL on NCM 460 cells and effect of 6-gingerol (10, 25, 50, 75, 100, 125, 150, 175 μg/mL) on Caco-2 cells were investigated. MTT assay was adopted to analyze cell proliferation. Methods of Gram staining and plate counting were used to analyze the probiotic adhesion to colonic epithelial cells. Results showed that 6-gingerol had no inhibitory effect on colonic epithelial cells within the non-cytotoxic dose range. 6-gingerol could promote the adhesion of probiotics to colon epithelial cells in a dose-dependent manner, which would be benefit for probiotics to exert continuous probiotic activity in the intestinal tract.

Abstract A leaf-disc-test method was used under controlled laboratory conditions to determine the feeding preference of Altica deserticola Latreille (Coleoptera: Chrysomelidae) on leaves of two liquorice species, Glycyrrhiza inflata Batalin and G. uralensis Fisch. ex DC. Leaf hardness and thickness, cuticle thickness, and nitrogen and tannin contents were compared between the two liquorices to explore their feeding resistance to A. deserticola. The larvae ate only G. uralensis leaves, while the adults fed on the leaves of both species but preferred those of G. inflata. The leaf hardness and thickness and cuticle thickness, as well as the nitrogen, total tannins, tannin chemicals contents in leaves, were significantly greater in G. inflata than in G. uralensis. The larvae having smaller chewing mouthparts could not feed on hard leaves with thick cuticle on both sides. The thicker cuticle and harder texture of G. inflata blades may be important physical traits for effective defence against larval phytophagy, while the higher tannin content in its leaves may be an important chemical trait determining their feeding preference. The larger adults, having stronger mouthparts, could consume nitrogen-richer G. inflata leaves to obtain the energy needed for flight and reproduction.

OBJECTIVES: Parkinson’s disease (PD) and the parkinsonian variant of multiple system atrophy (MSA-P) are distinct neurodegenerative disorders that share similar clinical features of parkinsonism. The morphological alterations of these diseases have yet to be understood. The purpose of this study was to evaluate gray matter atrophy in PD and MSA-P using regions of interest (ROI)-based measurements and voxel-based morphometry (VBM). METHODS: We studied 41 patients with PD, 20 patients with MSA-P, and 39 controls matched for age, sex, and handedness using an improved T1-weighted sequence that eased gray matter segmentation. The gray matter volumes were measured using ROI and VBM. RESULTS: ROI volumetric measurements showed significantly reduced bilateral putamen volumes in MSA-P patients compared with those in PD patients and controls (p<0.05), and the volumes of the bilateral caudate nucleus were significantly reduced in both MSA-P and PD patients compared with those in the controls (p<0.05). VBM analysis revealed multifocal cortical and subcortical atrophy in both MSA-P and PD patients, and the volumes of the cerebellum and temporal lobes were remarkably reduced in MSA-P patients compared with the volumes in PD patients (p<0.05). CONCLUSIONS: Both PD and MSA-P are associated with gray matter atrophy, which mainly involves the bilateral putamen, caudate nucleus, cerebellum, and temporal lobes. ROI and VBM can be used to identify these morphological alterations, and VBM is more sensitive and repeatable and less time-consuming, which may have potential diagnostic value.

Abstract The inhibitory effects of ginger essential oil (GEO) on chemically-induced cutaneous inflammation were investigated. Three kind of experimental murine models, such as xylene-induced ear edema model, carrageenan-induced paw edema model and 2,4-dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis (ACD) model were exploited to evaluate the anti-inflammatory effect of GEO. Results showed that GEO lowered the ear swelling rate induced by xylene and decreased paw edema rate induced by carrageenan of mice with cutaneous inflammation significantly. Treatment with GEO significantly inhibited DNFB-induced ear swelling degree and rate, and alleviated the inflammatory symptoms of ear skin lesions of ACD model mice markedly. Treatment with GEO decreased the gene expression of the inflammatory cytokines of auricular tissues such as TLR-2, TLR-4, TNF-α, IFN-γ, IL-1β, IL-8, and increased the gene expression of cytokine IL-4 significantly. In conclusion, GEO has anti-inflammatory and protective effect on chemically-induced cutaneous inflammation.

Abstract Streptococcus pneumoniae is one of the most frequent opportunistic pathogens worldwide. DNA processing protein A (DprA) is an important factor involved in bacterial uptake and DNA integration into bacterial genome, but its role in S. pneumoniae virulence remains unclear. The aim of this study was to characterize the effects of the pneumococcal dprA gene on the pathogenesis of S. pneumoniae. To construct a dprA-deficient pneumococcal strain, the dprA gene of the S. pneumoniae strain D39 was inactivated. The virulence of this dprA-deficient strain, designated ΔD39, was compared with that of the wild-type strain by evaluating their respective capabilities to adhere to human pulmonary epithelial cells (PEC-A549) and by analyzing their choline-binding protein expression levels. In addition, the expression profiles of genes associated with virulence and host survival assays were also conducted with the mutant and the wild-type strain. Our results indicate that the capability of ΔD39 to adhere to the PEC-A549 airway cells was significantly lower (p < 0.01) compared with D39. Additionally, the 100-KD choline-binding protein was not detected in ΔD39. The addition of competence-stimulating peptide (CSP) lead to a significantly reduction of psaA mRNA expression in the dprA-deficient mutant and an increased level of psaA transcripts in the wild-type strain (p < 0.01). The median survival time of mice intraperitoneally infected with ΔD39 was significantly higher (p < 0.01) than that of mice infected with D39. The results of this study suggest that DprA has a significant effect on virulence characteristics of S. pneumoniae by influencing the expression of choline-binding protein and PsaA.

ABSTRACT The risk of transfusion-transmitted malaria is a major concern in many countries. This study investigated the prevalence of malaria antibodies and parasitemia in eligible blood donors in Jiangsu, in Eastern China. Malaria antibodies were detected in 2.13% of the 704 plasma samples studied. We found that the prevalence of malaria antibodies was not significantly correlated with gender, occupation and frequency of donation, but it increased with age. No Plasmodium was observed in red blood cells and no Plasmodium DNA was detected in any of the antibody-positive samples. The prevalence of malaria antibodies was not higher than expected in Eastern China.

OBJECTIVES:Our study aimed to investigate the impact of fatigue on the severity of stroke and to explore the underlying mechanisms.METHODS:Fatigued male rats underwent middle cerebral artery occlusion and the infarcted brain area was determined. Then, coagulation parameters were assessed in the fatigued group and a control group. In addition, the level of fibrinogen was determined in rats deprived of sleep for various numbers of days. To study whether interleukin-6 was involved in fibrinogen synthesis during fatigue, we also measured levels of interleukin-6 in rats deprived of sleep for various numbers of days. Furthermore, brain injury by middle cerebral artery occlusion was measured in wild-type mice, interleukin-6-/- mice and wild-type mice treated with bezafibrate.RESULTS:More severe cerebral infarction was observed in the fatigued rats, resulting in an infarct ratio of 23.4%. The infarct ratio was significantly increased in the fatigued rats compared with that in the control group (8%, p<0.05). The level of fibrinogen was increased significantly in the fatigued rats compared with that in the control group. In addition, a marked reduction in fibrinogen level was observed in the fatigued interleukin-6-/- mice compared to their wild-type counterparts, whereas no difference was observed between fatigued wild-type mice and interleukin-6-/- rats treated with recombinant human interleukin-6. The reduction in brain injury due to middle cerebral artery occlusion during fatigue was observed in interleukin-6-/- mice and wild-type mice treated with bezafibrate.CONCLUSION:Fatigue could increase stroke severity and was associated with the interleukin-6-induced expression of fibrinogen.

Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.

The association of IGF-I gene polymorphisms with certain traits in 708 individuals of two Chinese dairy-goat breeds (Guanzhong and Xinong Saanen) was investigated. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing methods were employed in screening for genetic variation. Two novel mutations were detected in the 5'-flanking region and in intron 4 of IGF-I gene, viz., g.1617 G > A and g.5752 G > C (accession D26119.2), respectively. The associations of the g.1617 G > A mutation with milk yield and the body size were not significant (p > 0.05). However, in the case of g.5752 G > C, Xinong Saanen dairy goats with the CG genotype presented longer bodies (p < 0.05). Chest circumference (p < 0.05) was larger in Guanzhong goats with the GG genotype. In Xinong Saanen dairy goats with the CC genotype, milk yields were significantly higher during the first and second lactations (p < 0.05). Hence, the g.5752 G > C mutation could facilitate association analysis and serve as a genetic marker for Chinese dairy-goat breeding and genetics.

The plant Rhodiola crenulata is a perennial herbaceous species distributed in the plateau region of southwestern China, especially the Hengduan Mountains region. It has been one of the most important traditional herbal remedies in Tibet for more than one thousand years, but the accelerated and uncontrolled collection of this plant since the 1980s has lead to deforestation. We used inter-simple sequence repeats (ISSR) to assess levels of genetic variation in R. crenulata from nine diverse natural populations in eastern Tibet and northern Yunnan, the first time such a study has been carried out. The 12 primers we used were able to detect 184 polymorphic loc. Analysis of molecular variance (AMOVA) indicated that species level genetic diversity was relatively high (p = 97.83%, and Ho = 0.464) and analysis using Shannon’s index showed that the within and between genetic diversity of R. crenulata are approximately equal. Nei’s genetic distance and unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis showed that the three populations from Tibet and the six populations from Yunnan form two major clusters. The Yunnan populations from three locations were further divided into three corresponding groups, indicating that genetic differentiation was correlated to geographic distribution. Understanding the genetic structure of R. crenulata provides insight for the conservation and management of this endangered species.