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This study aimed to investigate the effect of the oral administration of sea cucumber protein (SCP) on wound healing.

The effects of soybean antigenic protein on the gustatory receptor T1R1 mRNA expression and intestinal immune response were evaluated in the obscure puffer (Takifugu fasciatus; initial weight 39.84 ± 3.09 g). Fish were fed with eight different diets for two months; four of these were practical diets with either a high level of fish meal (FMH 2 ), a low level of fish meal (FML 3 ), a diet where FML was partially (18.6%) replaced by fermented soybean meal (FMLF 4 ), and a diet where FML was partially (18.6%) replaced by soybean meal (FMLS 5 ). Four other semi-purified diets were offered that contained different levels (0%, 5%, 8%, and 12.5%) of soybean antigenic proteins (AP0, AP5, AP8, and AP12.5, respectively). Feeding rate and intestine histological structure were investigated and the expression of IL-1mRNA, IL-10mRNA, and TNFmRNA in the intestine, HSP70mRNA in the liver, and T1R1mRNA in the tongue and intestine in four practical-diet groups and two semi-purified-diet groups (AP0% and AP8%) were also determined. The obtained results show feeding rate increased gradually as levels of soybean antigenic protein among the practical-diet groups, while the feeding rate of semi-purified-diet groups (AP5-AP12.5%) decreased and then increased with increasing soybean antigenic protein level. The expression level of T1R1mRNA in the tongue was higher than in the intestine; while they showed the same tendency, FMLS group was significantly higher than the control FML, AP8 group was significantly higher than AP0 group, and significantly lower than FMLS group (containing 8.8% soybean antigenic proteins similar with AP8 group). Soybean antigenic proteins did not affect the width of the intestine folds or the thickness of the muscular layer; however, they caused a difference in the height of intestine folds: higher soybean antigenic protein levels in the semi-purified-diet group led to lower height of the intestine folds with mild injuries in the intestine. The expression level of IL-1mRNA, IL-10mRNA and TNFmRNA in the intestine showed the same tendency as T1R1mRNA; and the expression level of HSP70 of FMLS group in the liver was significantly lower than in the control FML. No significant difference was found among most indexes between FMLF and FML. In conclusion, single or interacting antigenic proteins and other anti-nutritional factors may change the expression of gustatory receptor T1R1mRNA in both the intestine and tongue by affecting the intestinal health, and thus affected feeding. Excessive soybean antigenic proteins resulted in intestinal damage, therefore fermented soybean meal is better for partly replacement of fish meal than unfermented soybean meal.

Based on the current driving recorder with satellite positioning function technical standards, a pure electric vehicle driving state data collection terminal is designed in this paper. Through the interface technology, the terminal reads batteries, motor speed, current, power and other information of pure electric vehicles that is different from traditional driving recorder data from traditional power vehicles, and then establish the corresponding monitoring platform. Comparing the content of telematics services with the traditional vehicles and pure electric vehicles, this paper illustrates the basic application and market of pure electric vehicle driving state data collection terminal.

Based on the technical standards for driving recorder with satellite positioning function, this paper designs a driving status data acquisition terminal for hybrid electric vehicle that could increase the data collection of battery, motor speed, current, power, etc. by the interface technology, and establishes appropriate monitoring platform at the same time. The test results show that, the driving status data acquisition terminal for hybrid electric vehicle meets to the application requirements.

Based on agricultural machinery body posture detection parameters and wheels gesture detection parameters collected by gyro inertial measurement unit, an agricultural machinery operation posture rapid detection method is proposed in this paper. The test results calibrated by RBF neural network show that, the test results of the method are accurate and available, and the method is effective and available for real-time body and wheel status data to further understand the agricultural machinery.

Based on agricultural machinery body posture detection parameters and wheels gesture detection parameters collected by gyro inertial measurement unit, an agricultural machinery operation posture rapid detection method is proposed in this paper. The test results show that, the test results of the method are accurate and available, and the method is effective and available for real-time body and wheel status data to further understand the agricultural machinery.

Designed a method based on the complexity of multiple nested parts to extract and analyze the information of parts. In order to simplify the algorithm and improve the efficiency of the program, a data structure stored all information of entities was presented. The theory of the region method was introduced in this paper, and a specific algorithm was designed to judge the nested relationship between contours by using this theory and entities information. Finally, the computational result can be used to determine the contour cutting order.

This study examined genetic variation in the major histocompatibility complex (MHC) Class II B gene in turbot (Scophthalmus maximus) by virulent bacterial pathogen challenge. One hundred fry from each of six families were infected with Edwardsiella tarda by intraperitoneal injection. Family mortality ranged from 28.0% to 83.3%. Complete exon 2 and intron 1 sequences of MHC Class II B genes were amplified from five survivor and five non-survivor individuals per family using the clone-sequence method. Thirty-seven sequences from 60 individuals revealed 37 different alleles, 25 of which were unique to this study. The 25 unique alleles belonged to 16 major allele types. Nine alleles were used to examine the association between alleles and resistance/susceptibility to disease. Five alleles were present in an individual, suggesting a minimum of three loci or copies of the turbot MHC Class II B gene. The rate of non-synonymous substitution (d N) was 2.30 and 1.58 times higher than synonymous substitution (d S) in the peptide-binding regions (PBR) and non-PBR in whole families, respectively, which suggested balancing selection on exon 2 of the MHC Class II B gene in turbot. One allele, Scma-DBB1*02, was significantly more prevalent in survivor stock than in non-survivor stock (P=0.001). Therefore, this allele might be associated with resistance to bacteria. A second allele, Scma-DBB1*10, was significantly more prevalent in non-survivor stock (P=0.021), and is likely associated with susceptibility to bacteria.

The half-smooth tongue sole (Cynoglossus semilaevis) is an important cultured marine fish as well as a promising model fish for the study of sex determination mechanisms. In the present study, a protocol for artificial induction of mitogynogenesis by hydrostatic pressure using heterologous sperm was developed in half-smooth tongue sole in order to assess homozygosity of gynogens and to identify WW super-female. The optimal initiation time for pressure shock of mitogynogenetic embryos was determined to be 21.5 min after insemination when water temperature is at 22-23°C, while the optimal pressure and treatment duration were determined to be 70 MPa for 4 min. About 1,500 mitogynogenetic diploid larvae were obtained. Ten tongue sole microsatellite markers were used for homozygosity analysis of 24 mitogynogenetic larvae. Among the 24 larvae, the percentage of homozygosity ranged from 73.91% to 87.50% with an average homozygosity of 80.54%. Sex-specific simple sequence repeat (SSR) markers, CseF-SSR1, were isolated and used for identifying WW super-female mitogynogens in the tongue sole. The amplification of genomic DNA using the sex-specific SSR marker produced one DNA band of 206 bp in ZZ males, two DNA bands of 206 and 218 bp in ZW females, and one DNA band of 218 bp in WW super-females. Four WW "super-female" gynogens were observed in 39 mitogynogenetic diploids, indicating a ZW sex determination mechanism in the tongue sole. Thus, a protocol for the induction of artificial mitogynogenesis has been developed for the first time in half-smooth tongue sole, and the WW super-female diploids were identified in the mitogynogens by sex-specific SSR markers. These findings lay the foundation and provide important tool for the elaboration of sex determination mechanism, generation of WW super-females, and development of clone line and breeding of all-female stock in the half-smooth tongue sole.

The paper first discusses the feasibility of the wireless communication in GIS (Gas Insulated Switchgear) substation. The signals inside GIS according to ISA100 standard are first classified and then a scheme to transmit some of the signals by ZigBee wireless is given. A demo example is shown to verify the proposed scheme. In the end, some future modifications and conclusion are discussed.

Pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH) are regulators of growth hormone secretion. In this article, we examined the difference in growth and mRNA expression of PACAP and GHRH between the sexes in half-smooth tongue sole, an important cultured fish species indicating sexually growth dimorphism in China. Firstly, a significant body weight difference between females and males was first observed at 7 months (P0.05) and at 18 onths the mean body weight of the females (771.0±44.3 g) was as much as 4.9 times higher than that of males (130.6±6.0 g). As a result, half-smooth tongue sole, Cynoglossus semilaevis, is a good model to investigate the effects of growth-related genes expression on sexual growth dimorphism. Secondly, the cDNAs encoding PRP/PACAP and GHRH were isolated. Two differently processed mRNA transcripts of PRP/PACAP (PRP-encoding and PRP splice variant) were found. PACAP and GHRH mRNA was highly abundant in brain and less abundant in other tissues. However, PACAP mRNA was expressed in most brain regions, and was lower in the cerebellum. GHRH mRNA was predominantly expressed in the hypothalamus and weakly expressed in all areas of the brain examined. Ontogenetic expression analysis indicated that PACAP and GHRH mRNA was detected in the early stages of embryogenesis. Finally, differential expression showed that there was no significant difference of the expression level of PACAP or GHRH between the sexes before 8 months of age. However, between 9 and 12 months of age, the GHRH mRNA expression level in males was significantly higher than in females (P0.05), which might be associated with GH deficiency in males. In contrast, the male PACAP mRNA expression level was not significantly higher than that in females even at 9 and 12 months of age. The present results provide important clues for understanding the sexual growth dimorphism mechanisms in half-smooth tongue sole.

In the present study, methodology of gynogenetic induction in spotted halibut were developed and optimized; the sex ratio of putative meiotic gynogenetic diploids was determined using AFLP-based molecular sexing technique; the homozygosity of gynogenetic population was assessed as opposed to cultivated population. The results showed that high percentage of meiotic gynogenetic diploids were generated when the eggs fertilized with irradiated heterologous sea perch frozen sperm (30–50 mJ cm−2) were cold shocked in sea water of −1°C for 40–75 min at 5 min after fertilization. About 15,200 diploid gynogenetic larvae were achieved and they exhibited normal morphology similar to diploid control. The gynogenetic diploids were 100% female, which first confirmed the female homogamete (XX/XY) sex determination in spotted halibut. The genetic analysis showed that the average H O was, respectively, 0.404 and 0.724 in gynogenetic population and cultivated population, indicating an increase of homozygosity in gynogenetic population.

Spotted Halibut (Verasper variegatus) is a commercially important marine fish species. In the present study, to isolate sex-specific markers in Spotted Halibut, we screened the genomes of Spotted Halibut by AFLP technique with 64 different primer combinations. Two primer combinations, MseI-CAG/EcoRI-ACC and MseI-CAT/EcoRI-AGG, produced a female-specific fragment in all females (n = 88) and in no males (n = 60, except 3 individuals), respectively. Both fragments were excised from the gel, cloned and characterized. The first fragment (named VevaF533) was 533 bp long, while the length of the second one (named VevaF218) was 218 bp. The two sequences showed no similarity to each other, and to the known sequences existing in the GenBank database using BLASTn. Cross-species amplification showed that the marker VevaF218 is a species-specific marker which is present in Spotted Halibut females but absent in Barfin Flounder (Verasper moseri). So this marker could be used for discriminating unambiguously between Spotted Halibut females and Barfin Flounder. Examination of the patterns of inheritance of VevaF218 in an interspecific hybrid family (V. variegatus ♂×V. moseri ♀) showed a female-specific pattern of inheritance from mother to daughter, implying that the marker VevaF218 is located on the female sex chromosome. This study provides a reliable AFLP-based genetic sexing of Spotted Halibut that could be useful for genetic mapping of the sex chromosomes and identification of sex-linked genes.

Effective methods for induction of gynogenetic diploids in Cynoglossus semilaevis are needed to initiate monosex culture. An effective protocol to induce half-smooth tongue sole gynogenesis using homologous sperm was developed in this study. A UV dose of 50 mJ cm⁻² was found to be the most effective for genetic inactivation of tongue sole sperm. Treatment optima for cold shocks were 5 °C for 20-23 min at 5 min after fertilization and the hatching rate of gynogenetic diploids was 10.0%. Microsatellite analysis at locus Csou 6 revealed that there was no genetic contribution from the paternal genome in 24 progenies of a meiotic gynogenetic family. Polymerase chain reaction demonstrated that only four individuals of 24 meiotic gynogenetic diploids produced the female-specific band of about 205 bp. The female/male ratio of gynogenetic diploids was significantly different from the theoretical ratio of 1:1. It is possible that there are some recessive lethal genes in W chromosome.

Half-smooth tongue sole females grow larger and faster than males. An all-female population would be of significant benefit for tongue sole aquaculture. In the present study, a female-specific AFLP marker (CseF305) was isolated from female genomic DNA of the tongue sole and sequenced. One pair of SCAR primers was designed based on the sequences of the female-specific marker. A PCR method for identifying genetic sex of the sole was developed. PCR amplification of genomic DNA from tongue sole adults using the SCAR primers resulted in a specific fragment in 30 female individuals, but not in 30 males. Secondly, effects of methyltestosterone treatment on the female:male sex ratio of tongue sole fry were examined. Methyltestosterone at a concentration of 20–100 μg/L·H 2 O can induce genetic females to reverse to phenotypic males in juvenile tongue sole and produce a high proportion of males (97–100% males). Phenotypic males with female genotype, that is, neo-males, were detected by using female-specific SCAR primers. The neo-males were cultured and matured, and used to mate with normal females to produce progeny. 130,000 fry were produced by using sperm from neo-males. Genetic sex identification demonstrated that 73% of the neo-male progeny fry contained female-specific DNA markers. Three combinations of sex chromosomes (ZZ, ZW and WW) were observed in the neo-male progeny. WW super-female individuals containing 2 huge heteromorphic chromosomes were found in some fry.

Emerging evidence have demonstrated that WISP1, a member of CCN protein family, plays an important role in the manifestation and development of many respiratory diseases, such as lung cancer, pulmonary fibrosis and asthma as well as ventilator-induced lung injury. The production of WISP1 and the following activation of WISP1-mediated Wnt signaling pathways may facilitate and even amplify the pathological processes of the diseases. Toll-like receptors and integrins are also participated in the signaling pathways. This review focuses on the impact and mechanism of WISP1 in pulmonary diseases and proposes that WISP1 holds promise as a diagnostic marker and/or therapeutic target.

Black snapper Sebastodes fuscescens (Houttuyn) is an economocally important species. Fourty-five microsatellite loci were isolated from an enriched genomic library of S. fuscescens. Ten of these loci were polymorphic in a test population with alleles per locus ranging from 2 to 5, and observed and expected heterozygosities per locus from 0.21 to 1.00 and from 0.19 to 0.86, respectively. Three loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. These polymorphic microsatellite loci would be useful for investigating genetic diversity of S. fuscescens and other related species.

Small yellow croaker (Pseudosciaena polyactis) is an economically important marine fish species. About 43 microsatellite loci were isolated from two enriched genomic library of Pseudosciaena polyactis. Ten of these loci were polymorphic in a test population with alleles per locus ranging from two to six, and observed and expected heterozygosities per locus from 0.3750 to 0.8750 and from 0.3112 to 0.8121, respectively. No loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found between pairs of loci. These polymorphic microsatellite loci would be useful for investigating genetic diversity of Pseudosciaena polyactis.

International audience; Genetic sex determination by W and Z chromosomes has developed independently in different groups of organisms. To better understand the evolution of sex chromosomes and the plasticity of sex-determination mechanisms, we sequenced the whole genomes of a male (ZZ) and a female (ZW) half-smooth tongue sole (Cynoglossus semilaevis). In addition to insights into adaptation to a benthic lifestyle, we find that the sex chromosomes of these fish are derived from the same ancestral vertebrate protochromosome as the avian W and Z chromosomes. Notably, the same gene on the Z chromosome, dmrt1, which is the male-determining gene in birds, showed convergent evolution of features that are compatible with a similar function in tongue sole. Comparison of the relatively young tongue sole sex chromosomes with those of mammals and birds identified events that occurred during the early phase of sex-chromosome evolution. Pertinent to the current debate about heterogametic sex-chromosome decay, we find that massive gene loss occurred in the wake of sex-chromosome 'birth'.

Background Genes in the major histocompatibility complex (MHC) have a critical role in both the innate and adaptive immune responses because of their involvement in presenting foreign peptides to T cells. However, the nature has remained largely unknown. Results We examined the genetic variation in MHC class IIB in half-smooth tongue sole (Cynoglossus semilaevis) after challenge with vibrio anguillarum. Two thousand and four hundred fry from 12 half-smooth tongue sole families were challenged with Vibrio anguillarum. To determine any association between alleles and resistance or susceptibility to V. anguillarum, 160 individuals from four high-resistance (HR, < 40.55% mortality) families and four low-resistance (LR, > 73.27% mortality) families were selected for MHC IIB exon2 gene sequence analysis. The MHC IIB exon2 genes of tongue sole displayed a high level of polymorphism and were discovered at least four loci. Meanwhile, the dN/dS [the ratio of non-synonymous (dN) substitutions to synonymous (dS) substitutions] in the peptide-binding region (PBR) was higher than that in the non-peptide-binding region (non-PBR). Eighty-eight alleles were discovered among 160 individuals, and 13 out of 88 alleles were used to analyze the distribution pattern between the resistant and susceptible families. Certain alleles presented in HR and LR with a different frequency, while other alleles were discovered in only the HR or LR families, not both. Five alleles, Cyse-DBB*6501, Cyse-DBB*4002, Cyse-DBB*6102, Cyse-DBB*5601 and Cyse-DBB*2801, were found to be associated with susceptibility to V. anguillarum with a frequency of 1.25%, 1.25%, 1.25%, 1.25% and 2.5% in the HR families, and 35%, 33.75%, 27.5%, 16.25%, 15% in the LR families (p < 0.01, 0.01, 0.01, 0.01, 0.01), respectively. Four alleles, Cyse-DBB*3301, Cyse-DBB*4701, Cyse-DBB*6801 and Cyse-DBB*5901, were found to be associated with resistance to V. anguillarum, with a frequency of 13.75%, 11.25%, 11.25%, 8.75% in the HR families and 1.25%, 1.25%, 1.25%, 1.25% and 1.25% in the LR families (p < 0.01, 0.05, 0.05 and p = 0.064), respectively. Conclusions Elucidation of the role of MHC II B genes in half-smooth tongue sole should prove to be helpful to the in-depth development of marker-assisted selective breeding in half-smooth tongue sole.

A large proteinolytic complex of M r ca 400 000 consisting of at least six distinct proteins, optimally active near pH 8, was purified from dry pea seeds using two different purification protocols. The better protocol used nonreducing buffers and was based on ammonium sulphate precipitation, anion exchange chromatography and preparative electrophoresis resulted in nearly a 2000-fold purification. Inclusion of reducing agents 2-mercaptoethanol or dithiothreitol appeared to modify the protease inhibitor sensitivity of the complex. A possible similarity of the particle to the mammalian proteasome is discussed.

The half-smooth tongue sole (Cynoglossus semilaevis, hereafter, “tongue sole”) is a marine flatfish with great commercial importance for fisheries and aquaculture in China. It has also been a promising model for the study of sex determination mechanisms in fish. Here, we report the construction of a genetic linkage map for the tongue sole, based on 137 markers including 103 AFLP markers, 33 microsatellite markers, and one female-specific DNA marker. Twenty-six linkage groups (LGs) were found. The total map length was 934.6 cM (Kosambi), with an average spacing of 8.4 cM, covering 64.4% of the estimated genome size. Furthermore, a female-specific SCAR marker, CseF-382, was mapped on LG5. This study represents the first genetic linkage map in the tongue sole. This map has great potential in the identification of quantitative traits loci and sex-related genes and marker-assisted selection in the tongue sole. Meanwhile, the new set of polymorphic microsatellite markers developed in this study is not only useful for genetic mapping but also of critical importance for studies on genetic diversity and broodstock management in tongue sole.

Half-smooth tongue sole (Cynoglossus semilaevis) is an important cultured marine fish as well as a promising model fish for the study of sex determination mechanisms. In the present study, a protocol for artificial gynogenesis of half-smooth tongue sole was developed in order to identify the sex determination mechanism and to generate all-female stock. The optimal UV-irradiation dose for genetically inactivating sea perch spermatozoa was determined to be > or =30 mJ/cm(2). The optimal initiation time for cold shock of gynogenetic embryos was determined to be 5 min after fertilization, while the optimal temperature and treatment duration were determined to be 20-25 min at 5 degrees C. Chromosomes from common diploids, gynogenetic haploids, and diploids were analyzed. WW chromosomes were discovered in some of the gynogenetic diploids. The microsatellite marker was applied to analyze gynogenetic diploid fry. Among the 30 gynogenetic diploid fry, 11 fry contained only one allele, while 19 contained two alleles, which had the same genotype as their mother. The female-specific DNA marker was observed in four individuals out of ten gynogenetic diploid fry. Ploidy analysis of 20 putative gynogenetic fry showed them all to be diploid. Thus, a protocol for the induction of artificial gynogenesis has been developed for the first time in half smooth tongue sole, and the sex determination mechanism in the tongue sole was determined to be female heterogametic with the ZW chromosome.

Experimental measurement and study of Raman spectra for pure ethanol and 50% ethanol aqueous solution has been conducted at ambient temperature and the pressures of 0.1-900 MPa by using Cubic Zirconia anvil cell. The result shows that the frequency of C-H stretch vibration increases with increasing pressure and each of their relation between the frequency and pressure can be expressed as following: for pure ethanol: v1 = 2881.890 + 0.00127 P + 6.213 x 10(-6) P2; V2 = 2928.707 + 0.004 38 P + 4.772 10(-6) p2; v3 = 2973.457 + 0.0089 P + 3.245 x 10(-6) p2; for 50% ethanol aqueous solution: v1 = 2885.616 + 0.010 8 P-2.699 x 10(-6) P2; v2 = 2932.734 + 0.0137 P - 3.346 x 0(-6) p2; v3 = 2978.115 + 0.0165 P - 4.914 x 10(-6) P2. Furthermore, the authors have observed that at the pressure lower than 550 MPa, the strength of hydrogen bonding becomes stronger with increasing pressure.