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A complete Dmrt4 DNA sequence was cloned from Cynoglossus semilaevis.The DNA sequence was 2 271 bp long,containing one intron and two exons.The NJ-phylogenetic tree indicated that the Dmrt4 gene of C.semilaevis had high homology to other fishes.Real-time PCR revealed that Dmrt4 was expressed in all seven tissues we examined in both sexes.Expression was significantly higher in the male pituitary than in other tissues.The expression of Dmrt4 in the gonads of methyl testosterone-treated males and neo-males was higher than in normal males.However,there was no significant difference between normal females and the high tempera-ture-treated neo-males.This indicates that Dmrt4 is not critical for sex reversal.Dmrt4 was expressed at signifi-cantly higher levels during the gastrula stage than during other stages.Dmrt4 was expressed at higher levels in males than in females.Our results provide a molecular basis for the formation of all female groups of C.semilaevis.

Using the microsatellite sequence got from genome sequencing,we constructed genetic linkage map with SSR markers for Japanese flounder(Paralichthys olivaceus),in which 681383B is the male parent and 6812E36 is the female parent.The male linkage map included 418 markers,which were distributed in 24 linkage groups,with 88.7% coverage.The length reached 1 418.1 cM,and the average marker interval was 3.62 cM.The female linkage map distributed in 24 linkage groups included 437 markers,with 89.1% coverage.spanning.The length reached 1 298.1 cM,and the average marker interval was 3.16 cM,.The medium density genetic linkage maps laid the groundwork for QTL analyses and marker-assisted selection breeding programs,and effectively promote the genetic improvement of Paralichthys,make flounder aquaculture sustainable development.

The pulse width is different, the mechanism of the laser-matter interaction is different. Damage results from plasma formation and ablation for10 ps and from heat depositing and conventional melting for ＞100 ps. Two theoretical models of transparent dielectrics irradiated by multi-pulses laser are respectively developed based on the above-mentioned different mechanism. One is the dielectric breakdown model based on electron density evolution equation for femtosecond multi-pluses laser, the other is the dielectric heat-damage model based on Fouriers heat exchange equation for nanosecond multi-pluses laser. Using these models, the effects of laser parameters and material parameters on the laser-induced damage threshold of dielectrics are analyzed. The analysis results show that different parameters have different influence on the damage threshold. The effect of parameters on the multi -pulses damage threshold is not entirely the same to the single-pulse damage threshold. The multi-pulses damage mechanism of dielectrics is discussed in detail, considering the effect of different parameters. The discussion provides more information for understanding its damage process and more knowledge to improve its damage thresholds. And the relationship between damage threshold and pulse number is illustrated, it is in good agreement with experimental results. The illustration can help us to predict the multi-pulses damage threshold and the lifetime of optical components.

With the development of the smart grid business management, the requirements of the information sharing and collaboration in power grid are increasing. An efficient application analysis system is needed to integrate and manage a variety of information in the multi-level, large, and complex power grid, including grid model, data, graphics and etc. An integrated power grid operation data center based on the research of big data is presented. The data center has been applied to a real engineering in China southern grid, and received good results. Big data technology widely used in the power industry will bring revolution, and lead the development of smart grid to a new stage.

With the development of additive manufacturing (AM), the constraints associated with traditional subtractive manufacturing have been largely alleviated. The improved design freedom due to the nature of additive manufacturing, which fabricates objects on a layer-by-layer basis, allows the manufacturing of parts of almost any shape theoretically. Apart from Design for Manufacturing (DMF), a new concept – Design for Additive Manufacturing (DFAM) has been introduced. One promising application arises from DFAM is the light weight structure, which can be achieved by topology optimization (TO). In this paper, two common TO methods will be reviewed. An important issue for applying TO to AM is that the optimized structure may not be simply and directly manufactured using an AM machine due to the constraints of AM technology. One particular constraint is the limited build size of most commercial 3D printers, which requires the part to be partitioned and printed in several parts. This limitation and some part partitioning methods outside TO will also be reviewed in this paper. Published version

The constantly increasing requirements of critical engineering parts can often be satisfied through the use of different materials with unique properties. In an effort to realize the potential of multi-functional parts, a study on the melt track characteristics of a copper based alloy (HOVADUR® K220) deposited on 316L steel via Selective Laser Melting (SLM) has been conducted. Single-line melt tracks, processed using 3 different sets of laser parameters, were printed on top of copper based alloy of variable height. It was found that the widths of the line tracks decreased with the height of the copper alloy. When processed at an energy density of 262J/mm3 the width of the copper alloy line tracks decreased from an average of 332μm to 151μm as the layers of underlying copper alloy increased from 0 to 40. Extensive cracking was observed when excessive energy density was used on layers 1 to 6. All scan tracks are continuous with minor amounts of unmelted particles. Track stability is observed to decrease slightly at higher layers as the track width decreases. NRF (Natl Research Foundation, S’pore) Published version

Female southern flounder( Paralichthys lethostigma) grow larger than males.Therefore,all-female production will maximize profit potential for the culture of this species.It may be effective to develop protocols to produce all-female southern flounder through induction of meiotic gynogenesis with the cryopreserved heterologous sperm of Lateolabrax japonicas.To test methods for inducing diploid gynogenesis in southern flounder using heterogenous sperm,the UV was used to inactivate sperm and pressure shock was used to prevent extrusion of the second polar body.The results of experiments showed that gynogenetic diploid can be induced by inactivated heterogenous sperm because that hybrid would die before hatching.Diploid gynogenesis was induced by activating egg development with UV irradiated sperm( 80 mJ/cm2) 4-5 min after fertilization ( 18 ℃) ,and then 65 MPa pressure treatment for 6 min.Flow cytometry and microsatellite DNA analysis were used to demonstrate that the gynogenetic fry are diploidy.These results indicate that the use of UV irradiated sperm from Lateolabrax japonicas for activation of flounder eggs and pressure shock for polar body retention is an effective method to produce gynogenetic diploid in southern flounder.

1 064 nm antireflection/532 nm high-reflection bichromatic coatings with and without barrier layer were fabricated on the UBK7 substrate using electron beam evaporation technique. The optical property and laser-induced damage threshold(LIDT) of the coatings were investigated, respectively. Compared with the coatings without barrier layer, the 1 064 nm LIDT of the bichromatic coatings with SiO2 barrier layer is 49.6% higher. By analyzing the morphology and depth information of laser-induced damage spots, and the standing wave electric field distribution of the film stacks, the mechanisms of 1 064 nm and 532 nm laser-induced damages of the bichromatic coatings were discussed. The results show that SiO2 barrier layer strengthens the coatings-substrate interface, and ther

Insulin-like growth factor-I (IGF-I) is a conserved peptide expressed ubiquitously,which shows highly homologous diverse effects on development,growth,and metabolism. With RT-PCR,the fragment encoding the turbot (Scophthalmus maximus) mature IGF-I peptide was amplified. It was predicted that the mature peptide was composed of 70 amino acids including 6 cysteines which may form 3 disulfide bonds. The target fragment was then successfully subcloned into the express vector pGEX-4T-1 and was highly expressed in E.coli BL21(DE3)plysS. The result of SDS-PAGE showed that the fusion protein expressed in the form of inclusion bodies with molecular weight of 34 ku and maximally amounted to 59 % of the whole protein in the E.coli cell 4 hours after being induced with IPTG. The western blotting indicated that recombinant protein had the antigenicity to anti-GST antibody. The inclusion bodies were dissolved in 6 mol/Lguanidine chloride followed by pulse renaturation in refolding buffer containing 0.5 mol/L L-Arginine,1.0 mol/L GSH and 0.2 mol/L GSSG. Then the renatured recombinant protein was purified by GSTrap FF affinity chromatography. The effect of purified GST-IGF-I on turbot kidney cells was analysed,and it indicated that the recombinant GST-IGF-I can stimulate the proliferation of the cells.

Half-smooth tongue sole(Cynoglossus semilaevis) is an important cultured marine fish and shows large sexual dimorphism in growth. In general,the females grow about four to five times faster than males. However,the percentage of males in cultured group is higher than that of females. Is the sex ratio deviation from expected value(1:1) associated with natural sex reversal? In the present study,the natural sex reversal phenomenon in half-smooth tongue sole was studied by means of female specific marker sex identification technique. We determined the phenotypic sex of 71 four-year-old half-smooth tongue sole by means of abdominal compression during reproduction season. The individuals not producing sperm or eggs were injected with (0.4-2.0) μg/kg LRHA3 and fed in concrete tanks until sexual maturation. In total,there were 39 phenotypic males and 32 females,and the mean body weight of phenotypic males was (0.339±0.078) kg and (1.471±0.214) kg of females. However,the body weight of one phenotypic male was 0.585 kg and significantly higher than other males. The female-specific marker analysis also showed that the phenotypic male tongue sole produced female-specific band,but not other 38 phenotypic males; 32 phenotypic females all produced female-specific fragment of about 205 bp. As a result,the phenotypic male half-smooth tongue sole producing female-specific fragment could be natural sex reversal male(neomale). Large-scale genetically sexual identification of phenotypic males in Laizhou Mingbo Co. Ltd showed the rate of natural sex reversal was 1.66%. It also proved that phenomenon of natural sex reversal in tongue sole was universal. Histological analysis of the sex-reversal male gonads showed that they contained functional testicular tissue with sprmatocytes. However,the amounts of spermatocytes in sex reversal individuals were fewer than that in control males. The phenomenon of natural sex reversal of female half-smooth tongue sole is helpful to explain the high male percentage in cultivating group and provides a new technique to produce all-female half-smooth tongue soles.

Leprosy is characterized by skin lesions and peripheral nerve damage. It may take a long time before the diagnosis can be confirmed if the patients have no typical skin involvements. Here we report an unusual case. A 40-year-old male with lepromatous leprosy showed a gradual onset of bilateral symmetrical neuropathies without characteristic skin manifestations seven years after onset and with pulmonary tuberculosis simultaneously. He was misdiagnosed as having Guillani-Barre syndrome and systemic necrotizing vasculitis for 10 years until the skin biopsy was performed. This case indicates that the risk of leprosy exists, though new cases being detected have significantly declined over the last 50 years; neurologists need to pay more attention to leprosy with various manifestations.

To investigate the allele polymorphism of MHC-DAA in Japanese flounder (Paralichthys olivaceus), specific primers were designed based on the published DAA-cDNA sequence of P. olivaceus to amplify the entire introns from flounder. Gene DAA consisted of four exons and three introns. The exon-intron structure was similar as that of Atlantic salmon. A polymorphic microsatellite was detected in intron 2. Subsequently, specific primer was designed based on the DAA genomic sequences. The genetic polymorphism within exon 2 of MHC-DAA was investigated by means of sequence analysis. Forty-five individuals were selected to amplify exon 2 and intron 2. One hundred and twenty-nine fragments revealed 30 novel alleles belonging to 18 allele major type. The frequency of alleles and the numbers of subtypes were not distributed equally. There were 55 variable sites in exon 2. The diversity of nucleotide within the sequence of exon 2 was 0.0887. The nucleotide sequence was corresponded with a putative peptide. Among the 83 amino acids in this peptide, 37.80% were variable, and 30 parsimony informative sites were observed. The rates of non-synonymous substitution (dN) were 3.30 and 2.43 times higher than the rate of synonymous substitution (dS) in the peptide-binding regions (PBR) and non-PBR, respectively. In this study, the high ratio of dN/dS value and high allelic diversity observed in flounder suggested a balancing selection in the exon2 of the MHC class IIA gene, which explained the high polymorphism of MHC-DAA gene in Paralichthys olivaceus.

To investigate the expression characteristics of sex related gene of FTZ-F1 in the half-smooth tongue-sole (Cynoglossus semilaevis), the homologue FTZ-F1 (hsFTZ-F1) full-length cDNA was isolated from the testis by homologous cloning, and the cDNA included the open reading frame and a 66 bp 5'-UTR, along with a 1 619 bp 3'-UTR, encoding a predicted 485 amino acid protein. Sequence, tissue distribution and phylogenic analyses of the FTZ-F1 showed that the hsFTZ-F1 belonged to SF-1/Ad4BP group. The hsFTZ-F1 transcripts were highly abundant in the gonads, kidneys, brain and head-kidneys, but weakly in other tissues. However, the expression level in the brain and head-kidney of female was highly abundant than in the male. The hsFTZ-F1 expression was highly abundant in the embryo than in the larvae, which suggested that the hsFTZ-F1 may be involved in the organogenesis in the tongue sole.

This work provides the results for W incorporated diamond-like carbon films with thickness from 86 to 110 nm prepared by the pulsed-laser deposition method. Atomic force microscope (AFM), X-ray diffractometer (XRD), Raman spectroscopy and ellipsometry were used to characterize the films. The surface images indicated that the surface roughness (RMS) varied from 1.15 to 4.03 nm and was not sensitive to the W incorporation. α-W 2 C and WC phase were found to form, which could be attributed to the W incorporation. The complex index of refraction almost linearly changed with the increase in W deposition time, while the extinction coefficient exhibited a complicated behavior. By the analysis of the Raman spectra, it was found that no obvious affection was induced after the W incorporation.

The problem of mixed H2/H∞ filtering for polytopic Delta operator systems is investigated. The aim is to design a linear asymptotically stable filter which guarantees that the filtering error system has different performances in different filtering channels. Based on a parameter-dependent Lyapunov function, a new mixed H2/H∞ performance criterion is presented. Upon this performance criterion, a sufficient condition for the full-order mixed H2/H∞ filter is derived in terms of linear matrix inequalities. The filter can be obtained from the solution of a convex optimization problem. The proposed filter design procedure is less conservative than the strategy based on the quadratic stability notion. A numerical example is given to illustrate the feasibility of the proposed approach.