Chemical fingerprint analysis of rhizomes of Gymnadenia conopsea by HPLC-DAD-MSn

A high-performance liquid chromatography-diode array detection-tandem mass spectrometry (HPLC-DAD-MS(n)) method has been firstly developed for chemical fingerprint analysis of rhizomes of Gymnadenia conopsea R. Br. and rapid identification of major compounds in the fingerprints. Comparing the UV and MS spectra with those of reference compounds, seven main peaks in the fingerprints were identified as adenosine (1), 4-hydroxybenzyl alcohol (2), 4-hydroxybenzyl aldehyde (3), dactylorhin B (4), loroglossin (5), dactylorhin A (6) and militarine (7). Compounds 4-7 were succinate derivative esters and firstly discovered from this species. The Computer Aided Similarity Evaluation System (CASES) for chromatographic fingerprint of traditional Chinese medicine was employed to evaluate the similarities of 10 samples of the rhizomes of G. conopsea collected from Sichuan, Qinghai and Hebei provinces, Tibet autonomous region of China, and Nepal. These samples from different sources had similar chemical fingerprints. This method is specific and may serve for quality identification and comprehensive evaluation of this traditional Tibetan remedy.     

Genomic Selection Using BayesCπ and GBLUP for Resistance Against <Emphasis Type="Italic">Edwardsiella tarda</Emphasis> in Japanese Flounder (<Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>)

The Japanese flounder is one of the most widely farmed economic flatfish species throughout eastern Asia including China, Korea, and Japan. Edwardsiella tarda is a major species of pathogenic bacteria that causes ascites disease and, consequently, a huge economy loss for Japanese flounder farming. After generation selection, traditional breeding methods can hardly improve the E. tarda resistance effectively. Genomic selection is an effective way to predict the breeding potential of parents and has rarely been used in aquatic breeding. In this study, we chose 931 individuals from 90 families, challenged by E. tarda from 2013 to 2015 as a reference population and 71 parents of these families as selection candidates. 1,934,475 markers were detected via genome sequencing and applied in this study. Two different methods, BayesCπ and GBLUP, were used for genomic prediction. In the reference population, two methods led to the same accuracy (0.946) and Pearson’s correlation results between phenotype and genomic estimated breeding value (GEBV) of BayesCπ and GBLUP were 0.912 and 0.761, respectively. In selection candidates, GEBVs from two methods were highly similar (0.980). A comparison of GEBV with the survival rate of families that were structured by selection candidates showed correlations of 0.662 and 0.665, respectively. This study established a genomic selection method for the Japanese flounder and for the first time applied this to E. tarda resistance breeding.     

An Excitatory Neural Assembly Encodes Short-Term Memory in the Prefrontal Cortex

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The superior fault tolerance of artificial neural network training with a fault/noise injectionbased genetic algorithm

Artificial neural networks (ANNs) are powerful computational tools that are designed to replicate the human brain and adopted to solve a variety of problems in many different fields. Fault tolerance (FT), an important property of ANNs, ensures their reliability when significant portions of a network are lost. In this paper, a fault/noise injection-based (FIB) genetic algorithm (GA) is proposed to construct fault-tolerant ANNs. The FT performance of an FIB-GA was compared with that of a common genetic algorithm, the back-propagation algorithm, and the modification of weights algorithm. The FIB-GA showed a slower fitting speed when solving the exclusive OR (XOR) problem and the overlapping classification problem, but it significantly reduced the errors in cases of single or multiple faults in ANN weights or nodes. Further analysis revealed that the fit weights showed no correlation with the fitting errors in the ANNs constructed with the FIB-GA, suggesting a relatively even distribution of the various fitting parameters. In contrast, the output weights in the training of ANNs implemented with the use the other three algorithms demonstrated a positive correlation with the errors. Our findings therefore indicate that a combination of the fault/noise injection-based method and a GA is capable of introducing FT to ANNs and imply that the distributed ANNs demonstrate superior FT performance.     

Locus Mapping,Molecular Cloning,and Expression Analysis of <Emphasis Type="Italic">rps6kb2</Emphasis>, a Novel Metamorphosis-Related Gene in Chinese Tongue Sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>)

Flatfish metamorphosis denotes the extraordinary transformation of a symmetric pelagic larva into an asymmetric benthic juvenile. This unique process involves eye migration, a 90° rotation in posture, and asymmetrical pigmentation for adaptation to a benthic lifestyle. In the present study, we used genetics to map a metamorphosis-related locus (q-10M) in the male linkage group (LG10M), a small interval of 0.9 cM corresponding to a 1.8 M-bp physical area in chromosome 9 in the Chinese tongue sole (Cynoglossus semilaevis). Combined with single-marker analysis, ribosomal protein S6 kinase 2 (rps6kb2) a member of the family of AGC kinases was identified as a novel metamorphosis-related candidate gene. Its expression pattern during metamorphosis was determined by quantitative RT-PCR and whole-mount in situ hybridization analysis. rps6kb2 gene was significantly expressed in metamorphic climax stage larvae and distributed in all the tissues transforming during metamorphosis, including tail, jaw, eye and skin of larvae. The results suggest that rps6kb2 has a general role in tissue transformations during flatfish metamorphosis including tail changes, skull remodeling, eye migration, and asymmetrical pigmentation.     

Investigation of Antimicrobial Resistance in Escherichia coli and Enterococci Isolated from Tibetan Pigs

Objectives

This study investigated the antimicrobial resistance of Escherichia coli and enterococci isolated from free-ranging Tibetan pigs in Tibet, China, and analyzed the influence of free-ranging husbandry on antimicrobial resistance.

Methods

A total of 232 fecal samples were collected from Tibetan pigs, and the disk diffusion method was used to examine their antimicrobial resistance. Broth microdilution and agar dilution methods were used to determine minimum inhibitory concentrations for antimicrobial agents for which disks were not commercially available.

Results

A total of 129 E. coli isolates and 84 Enterococcus isolates were recovered from the fecal samples. All E. coli isolates were susceptible to amoxicillin/clavulanic acid, and 40.4% were resistant to tetracycline. A small number of isolates were resistant to florfenicol (27.9%), ampicillin (27.9%), sulfamethoxazole/trimethoprim (19.4%), nalidixic acid (19.4%), streptomycin (16.2%) and ceftiofur (10.9%), and very low resistance rates to ciprofloxacin (7.8%), gentamicin (6.9%), and spectinomycin (2.3%) were observed in E. coli. All Enterococcus isolates, including E. faecium, E. faecalis, E. hirae, and E. mundtii, were susceptible to amoxicillin/clavulanic acid and vancomycin, but showed high frequencies of resistance to oxacillin (92.8%), clindamycin (82.1%), tetracycline (64.3%), and erythromycin (48.8%). Resistance rates to florfenicol (17.9%), penicillin (6.0%), ciprofloxacin (3.6%), levofloxacin (1.2%), and ampicillin (1.2%) were low. Only one high-level streptomycin resistant E. faecium isolate and one high-level gentamicin resistant E. faecium isolate were observed. Approximately 20% and 70% of E. coli and Enterococcus isolates, respectively, were defined as multidrug-resistant.

Conclusions

In this study, E. coli and Enterococcus isolated from free-ranging Tibetan pigs showed relatively lower resistance rates than those in other areas of China, where more intensive farming practices are used. These results also revealed that free-range husbandry and absence of antibiotic use could decrease the occurrence of antimicrobial resistance to some extent.     

藏药全缘叶绿绒蒿的化学成分研究

从藏药全缘叶绿绒蒿(Meconopsis integrifolia(Maxim.)Franch)全草乙醇提取物中分离得到六个化合物,分别鉴定为:普托品碱(protopine,1)、马齿苋酰胺E(Oleracin E,2)、木犀草素(luteolin,3)、二氢槲皮素(dihydro quercetin,4)、洋芹素(apigenin,5)和小麦黄素(tricin,6),其中,化合物2～6为首次从该植物中分离得到.     

Determination of Benzalkonium Chloride Partition in Micelle Solutions Using Ultrafiltration Method

The objectives of this study were to determine the concentrations of free benzalkonium chloride (BAC) and apparent partitions coefficients (K _m) in micelle solutions and to explore its application in formulation development. Ultrafiltration (UF) was carried out using 10K Nanosep® devices and centrifugation at 5,000 rpm for 5 min. The separation of free BAC from micellar solutions was also conducted using ultracentrifugation (UC) method for the comparison with UF method. Capillary electrophoresis method was used for the identification of micelles. Results showed that a UF method was applicable for quantitatively evaluating BAC–micelle interaction in micellar solutions. Unlike UF, UC could not completely separate free BAC from the micelles. The free BAC concentrations in the micelle solutions decreased with increasing surfactant concentrations. Among polysorbate 80, cremophor EL, and tyloxapol, BAC had the highest K _m in polysorbate 80 solutions. The K _m was significantly lower in non-buffered aqueous solutions than that in citric buffers. Moreover, increasing surfactant concentrations led to reducing antimicrobial activity. The UF is a rapid and accurate method that minimally alters the micellar equilibrium for the determination of free BAC and K _m in micellar solutions. In conclusion, free BAC concentration, which is a function of surfactant type, surfactant concentration, and ion strength of solution, is likely associated with the antimicrobial activity.     

Prevalence,Associated Risk Factors,and Phylogenetic Analysis of Toxocara vitulorum Infection in Yaks on the Qinghai Tibetan Plateau,China

Toxocara vitulorum has been rarely reported in yaks at high altitudes and remote areas of Sichuan Province of Tibetan Plateau of China. The current study was designed to investigate the prevalence, associated risk factors, and phylogenetic characteristics of T. vitulorum in yak calves on the Qinghai Tibetan plateau. Fecal samples were collected from 891 yak calves and were examined for the presence of T. vitulorum eggs by the McMaster technique. A multivariable logistic regression model was employed to explore variables potentially associated with exposure to T. vitulorum infection. T. vitulorum specimens were collected from the feces of yaks in Hongyuan of Sichuan Province, China. DNA was extracted from ascaris. After PCR amplification, the sequencing of ND1 gene was carried out and phylogenetic analyses was performed by MEGA 6.0 software. The results showed that 64 (20.1%; 95% CI 15.8–24.9%), 75 (17.2; 13.8–21.1), 29 (40.9; 29.3–53.2), and 5 (7.6; 2.5–16.8) yak calves were detected out to excrete T. vitulorum eggs in yak calve feces in Qinghai, Tibet, Sichuan, and Gansu, respectively. The present study revealed that high infection and mortality by T. vitulorum is wildly spread on the Qinghai Tibetan plateau, China by fecal examination. Geographical origin, ages, and fecal consistencies are the risk factors associated with T. vitulorum prevalence by logistic regression analysis. Molecular detection and phylogenetic analysis of ND1 gene of T. vitulorum indicated that T. vitulorum in the yak calves on the Qinghai Tibetan plateau are homologous to preveiously studies reported.     

Bifidobacteria alleviate experimentally induced colitis by upregulating indoleamine 2, 3‐dioxygenase expression

The goal of this study was explore the role of indoleamine 2, 3‐dioxygenase (IDO) in the therapeutic effect of probiotics on inflammatory bowel disease (IBD). Trinitrobenzene sulfonic acid (TNBS) was used to induce colitis in mice and 1‐methyltryptophan (1‐MT) to block expression of IDO. Clinical manifestations and macroscopic and microscopic colonic changes were assessed using a disease activity index (DAI), the Wallace–Keenan, and Curtner scoring systems, respectively. Expression of colonic IDO was detected by western blot. Immunohistochemistry analysis to evaluate numbers of CD11c⁺ cells and expression of IL‐17 and Foxp3 showed that DAI, Wallace–Keenan, and Curtner scores were lower in the Bifidobacteria treatment group than the control group and that the therapeutic effect of Bifidobacteria was blocked by 1‐MT (P < 0.05). Additionally, Bifidobacteria were found to increase expression of IDO and the numbers of CD11c⁺ cells, CD11c⁺ and IDO double positive cells and Foxp3⁺ Treg cells, while decreasing the number of IL‐17⁺cells (P < 0.05). The generation of Foxp3⁺ Treg cells induced by Bifidobacteria was abrogated by 1‐MT (P < 0.05). These findings study suggest that Bifidobacteria attenuate TNBS‐induced colitis by inducing expression of IDO, which further increases generation of Foxp3⁺ Treg cells.