Abstract:MicroRNAs ( miRNAs) is singlestranded oligoribonucleotides about 20 - 25 nucleotides ( nt) in length and function at many biological process,for example,cell proliferation,apoptosis,growth,differentiation and metabolism. MiR-27 play an important role in adipogenesis and muscle stem cell behavior,but it is unclear about its role in the development of neural cell. In the present study,the target genes of miR-27 was analyzed by miRBase and TargetScan databases. A recombinant miR-27 eukaryotic expression vector was constructed and expressed it in human glioma cells U343. We reconstructed the firefly luciferase and renilla luciferase vector and fused the 3′-UTR of miR-27 target gene Bmi1 with report gene that was transfected in U343 cells. luciferase activity was detected by Dual-Luciferase Reporter ( DLR) Assay. Results showed the common target genes of miR-27a and miR-27b regulated mainly the process of development. MiR-27 eukaryotic expression vector can produce mature miR-27a and miR-27b. Firefly luciferase activity was significantly reduced by cotransfection of miR-27a (P < 0. 05),miR-27b (P < 0. 05),combination of both (P < 0. 01) and sense 3′-UTR of Bmi1. These results implied that Bmi1 may be the one of miR-27 target genes.

Abstract:The Razor Clam (Sinonovacula constricta)is an important shellfish in China,Japan and Korea. We sequenced mitochondrial cytochrome oxidase Ⅰ ( CO Ⅰ) genes of 197 individuals from nine populations to investigate population genetic structure of the northern (ZH,HG,HY),the mid-regional(YC,DT,BZ) and the southern(NB,TZ,ND)groups of S. constricta along the coast of China. We identified 125 haplotypes and 96 variable sites from 556 bp of CO Ⅰ. Nucleotide diversity ranged from 2. 176 4 to 7. 497 0. Analysis of molecular variance ( AMOVA) showed that interpopulation variance accounted for 80. 27% ( P < 0. 05 ), whereas the remaining 18. 74% and 0. 99% were detected within population and among populations of same groups respectively,indicating significant genetic differentiation among populations. Phylogenetic tree analysis and genetic distance analysis showed that the nine populations were clustered into three distinct groups,with the northern group clustering first with the southern group and then with the mid-regional group.

Abstract:The purpose of this study is to clarify the function of mitogen-activated protein kinase ( MAPK) activity in the reinitiation of the first meiosis in Zebra Fish (Danio rerio) oocytes. The oocyte maturation of D. rerio was induced in vitro by 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DHP). Another group of oocytes were treated by 17α,20β-DHP with U0126,the inhibitor of MAPK phosphorylation. MAPK phosphorylation in oocytes obtained at different culture time points was analyzed with Western Blot, and germinal vesicle breakdown (GVBD) of oocytes was observed. The results showed that MAPK activity in oocytes in response to the 17α,20β-DHP induction ( without U01266 ) was significantly increased at 2 h of culture and was maintained at a high level until 6 h,and GVBD also almost completed at 6 h. In contrast,U0126 completely blocked MAPK phosphorylation in oocytes exposed to 17α,20β-DHP. GVBD rate of oocytes cultured with both of 17α,20β-DHP and U0126 was lower than that of oocytes cultured with only 17α,20β-DHP. The results suggest that MAPK participates in the resumption of the first meiosis in D. rerio oocytes.

Abstract:The structure and related substitution rate of mitochondrial DNA displacement loop ( D-loop) region and cytochrome b ( Cyt b) were comparatively analyzed through Bayesian MCMC simulation in present study. Based on the sequence analysis of mtDNA D-Loop 648-680 bp,the D-Loop structure of this species is similar to that of other fish species containing the extended termination associated sequence domain ( ETAS),the central conserved domain ( CD) and several conserved sequence block ( CSB-F、CSB-E、CSB-D、CSB-1 ) .According to comparative analysis of related substitution rate in Neosalanx taihuensis,the rate of polymorphic loci was lower in D-Loop (0. 83% ) than in Cyt b (1. 31% ) and the interval estimation of related substitution rate is 1. 000 ± 0. 131 in Cyt b and 0. 859 ± 0. 261 in D-Loop,respectively,which indicated that the mutation rates of D-loop were slower than that of Cyt b with large variance of Bayes posterior density distribution in N. taihuensis. It provided evidence that the substitution rate of Cyt b is higher than that of D-Loop,and is more approximate to molecular clock hypothesis,which indicated that Cyt b might be more appropriate molecular marker than D-Loop in studies on molecular ecology and phylogeography for this species.

Abstract:The breeding ecology of Common Tern (Sterna hirundo) and Gull-billed Tern (Gelochelidon nilotica) were studied in Hongjiannao Lake of Shaanxi Province from April to July of 2009 by direct observation and by counting the nests. Both of them arrived at Hongjiannao Lake in late April and bred in the middle of May. Both species built their hest at the islands in Hongjiannao Lake,but the nest-sites of S. hirundo located in the margin area of the islands in linear patterns,and the nest-sites of G. nilotica were built in the center region in a patch patterns. Food items of the two species were partially separated. Therefore,the separations in nest-sites and food items were regarded as the major reasons for the two identical bird breeding in the same area. The growth curves of the nestling body mass of the two species matched with the description of Gompertz equation and the difference in growth rate of the two birds might reduce the interspecies competition.

Abstract:Breeding habit of Red-billed Leiothrix ( Leiothrix lutea) was preliminarily studied at Longmenhe Village of Xingshan County,Hubei from April,2005 to August,2007. L. lutea reproduce during the middle ten days of April to the last ten days of July in Longmenhe area. The birds built their nests on the Dwarf Bamboo (Indocalamus wilsoni) or short shrub shaded by arbor or shrubbery on slope or near the farmland and footpath. The building materials include withe,slender root,bine,dwarf bamboo leaves,grass leaves and stems,and lichen and so on. The process of building nest last 5 to 7 days and both male and female were involved in. Female lay an egg in early morning everyday. The mean clutch size is 3. 41 ± 0. 80 ( n = 61) . The altricial nestlings hatched after 11-13 d (n = 6) incubation. The parent fed their offspring for 9 or 10 days before they left nests when they have not full-developed yet. The breeding successful rate is about 22. 95% ( n = 61) in the study area.

Abstract:From July 2008 to February 2009,we conducted a survey on the bats species diversity in 47 caves at 14 townships in eastern of Songtao County,Guizhou Province. We found it showed 14 species belonging to 3 families,one of them need to identify further. Three species, Pipistrellus javanicus, P. paterculus, and Rhinolophus monoceros are the new records to Guizhou Province,and R. monoceros was found first time in the mainland of China. We also collected information on the hibernation behavior and habitat selection of bats,as well as the threats to their habitat. The hibernation behavior was obvious differences between different species of bat,and the species variation between caves might be indicated by the habitat type difference out of the cave.

Abstract:Primers were designed according to the GenBank sequence of FSHβ gene of the homologous species of Cricetulus barabensis. Partial sequence of exon 3 in FSHβ gene was cloned by PCR method from random samples of C. barabensis in Yinan,Linqu and Qufu,Shandong Province. It is 775 bp and its accession number in GenBank is GQ456067. The homologies of nucleotide and amino acid are 80%-96% and 79%-100% respectively when compared with the same region sequence of Homo sapiens,Rattus norvegicus,Mus musculus, Pan troglodytes,and Ovis aries. The result of the phylogenetic evolution analysis by FSHβ sequence is in consistent with the relative species relationship,and therefore the sequence of FSHβ gene could be used as an ideal marker to study the species relationships or hereditary distance.

Abstract:Diets of Black-faced Spoonbill (Platalea minor) were mainly fish and shrimps. Survey on species of fish and shrimps in the feeding habitat of the spoonbill would be helpful to analyze the potential prey of this endangered species. Xinhua Bay in Fujian province is one of the most important wintering sites of Black-faced Spoonbill in mainland China. Wintered Black-faced Spoonbills in Xinhua Bay usually rest at the aquaculture ponds in day time,while feed at the intertidal mudflats which are about 8 km away from the farm at night. Forty two species of fish and 6 species of shrimps were collected in the feeding sites. According to the body size, habit,distribution habitat and number of them,19 species of fish and 6 species of shrimps would be the potential prey of Black-faced Spoonbill,with Mugilidae being the most important food resource. The protection of fish and shrimps in intertidal mudflats should be done to promote the conservation of Black-faced Spoonbill.

Abstract:The Wetland Reserve located on the west side of the Taipa-Coloane Causeway is an important wintering area for the Black-faced Spoonbill ( Platalea minor). The population size of Black-faced Spoonbill wintering at the Wetland Reserve has increased annually. Surveys conducted in the winters from 2007 to 2009 indicated that the Black-faced Spoonbill wintered in Macao from early November to April. Wintering populations were largest from December to February,exceeding 50 individuals. The artificial wetland is the primary rest areas for Black-faced Spoonbills,while tidal flats serve as important feeding-grounds. Daytime behavior consists mainly of resting,followed by maintenance. Traffic noise is the main human disturbance around the Wetland Reserve,but the level of noise was relatively low. The predicted value of traffic noise at the Black-faced Spoonbills’habitat were 45. 4 dB( A) and 46. 5 dB( A),below the threshold of avian tolerance. Most alarm behaviors were caused by large tractors and helicopters. With rapid infrastructure development around the Wetland Reserve,traffic flow will be further increased. Controlling the surrounding traffic flow and adjusting the flight course of helicopters to detour around the airspace at the Wetland Reserve is suggested as part of a management plan for decreasing disturbance on wintering Black-faced Spoonbills at the Wetland Reserve of Macao.

Abstract:It is difficult to identify the sex of Ostrich ( Struthio camelus) with morphological method before reaching sexual maturity,and this situation challenged the early feeding of Ostrich. In this study,genome DNA was extracted from feathers by using nondestructive sampling method and a combination of primer from one of EE0. 6 and the other of CHD. Sex gene sequences were amplified and compared among three pairs of sex-known birds and 9 sex-unknown birds. The results showed that male birds displayed no distinct amplification band, whereas one band was amplified for female ones. Thus the method could be used for sexing Ostrich before sexual maturity.

Abstract:The life history and main development characteristics of different development stages of Craspedacusta xinyangensis were recorded. Under the condition of 25-28℃ ,the eggs would develop into 0. 14-0. 21 mm long,stick shape planula 23 h after fecundation. Under the experimental condition,The planula would float in the water for 3-5 days and then stayed on the bottom of the glass container. In the following 3-4 days,they would develop into a 0. 3-0. 6 long hydranth. The hydranth would reproduce unciliated frustule which would reproduce new hydranth. After the hydranth reached the maturation, only one medusa bud would be reproduced. The newly released young medusa had 16 tantacle.

Abstract:The aim of this study was to investigate mitochondrial 16S rRNA sequence variation to assess genetic structure and polymorphism of in four stocks of Marsupenaeus japonicus. The samples of M. japonicus were obtained from the East Sea and South Sea of Guangdong ( GD),Taiwan ( TW),Fujian ( FJ) and Zhejiang (ZJ) Province in China. The PCR technique was used to amplify the mtDNA 16S rRNA gene fragment. Four hundred and seventy bp nucleotide sequence of partial 16S rRNA was obtained,and 16 variable sites and 10 haplotypes were detected among all sequences. Nucleotide diversities in GD stock,TW stock,FJ stock and ZJ stock were 0. 000 8,0. 001 0,0. 005 1 and 0. 001 5 respectively. Each stock had shared haplotype and unique haplotype. F_st analysis showed that genetic difference existed among four stocks,and significant genetic differentiations were observed between FJ stock and other three stocks. The Kimura-2-paramter genetic distance calculated by the MEGA software between the stock of FJ and TW was the highest,up to 0. 003 2,while that between the stock of ZJ and GD was the lowest to 0. 000 9. NJ phylogenetic tree was constructed using 12 Penaeidae shrimp. These results will be important and useful for making scientific strategy for the family selective breeding and molecular marker assisted selective breeding.

Abstract:The karyotypes of three species of Tetraodontiformes fishes were analyzed. The PHA and colchicine were injected in vivo,and kidney cells were collected and treated with lower osmotic pressure and air-drying technique. The results showed that these three marine species had diploid chromosome karyotypes,and all did not contain satellite chromosome,secondary constrictions and sex chromosome. The karyotype formula were as follows: the karyotype of Monacanthus chinensis was 2n = 34,34t,NF = 34; the karyotype of Takifugu xanthopterus was 2n = 44,12m + 8sm + 24t,NF = 64; and the karyotype of T. rubripes was 2n = 44,14m + 6sm + 24t,NF = 64; We also compared the karyotypes of the related fishes in the order of Tetraodontiformes.

Abstract:Insulin-like growth factor 2 gene was one of important genes controlling muscle growth and fat deposition in animals. In this study,we isolated 5 475 bp IGF2 including complete coding sequence 669 bp which interrupted by 3 introns and encoded 222 amino acids from GIFT strain Nile Tilapia Oreochromis niloticus. Total eleven SNPs were found at GIFT strain Nile Tilapia O. niloticus IGF2 exons and introns through comparing sequences from 10 individuals. The genetypes of intron 1 621 nt and exon 3 161 nt were studied and the correlation of fish shape,weight gain with different genetypes were analyzed in this paper. Tetra-primers ARMS was performed to detect the SNP locus at 621 nt ( C /T) of intron 1. The results indicated that the frequencies of CC,CT and TT were 0. 32,0. 32 and 0. 36 in male population,and 0. 38,0. 38 and 0. 24 in female population respectively; there was only a remarkable correlation between different genotypes with male GIFT strain Nile tilapia O. niloticus body shape (height /length) (P < 0. 05),CC individuals were significantly higher than CT and TT individuals. The SNP locus at 167 nt ( A /G) of exon3 was a nonsense mutation but it changed the restriction site of MSP Ⅰ. So PCR-RFLP was used to detect the genotypes,only GG and AG individuals were found,with the frequencies of GG and AG 0. 71 and 0. 19 in male population and 0. 75 and 0. 25 in female population respectively; there was only a remarkable correlation between different genotypes with male GIFT strain Nile tilapia O. niloticus weight gain ( P < 0. 01),GG individuals were obviously higher than AG ones.

Abstract:The genetic diversity and sequence variation of mtDNA D-loop in the population of vulnerable species Onychostoma lini from the Duliu River in Guizhou were studied for the first time on the basis of determination of about 470 bp sequences of mtDNA D-loop in 36 individuals by the methods of PCR and DNA sequencing. The results showed that the length of the determined sequence in 36 individuals varied from 469 to 475 bp,and the total percentage of bases A and T (68. 4% ) was more than that of bases G and C (31. 6% ) in the sequence. A total of 25 polymorphic loci,including 19 transitions and 6 transversions,were detected in the sequence of the population. The nucleotide diversity (π) of the 36 individuals was 0. 005 75,and the average number of nucleotide differences (K) of them was 2. 695. The 36 individuals belonged to 5 haplotypes according to the determined sequences. The haplotypic diversity ( Hd) was 0. 260,and the average genetic distance ( P) between the haplotypes of the population was 0. 026. The UPGMA phylogenetic tree of the 5 haplotypes comprised 2 branches. The data show that the population of O. lini in the Duliu River has abundant variation in mtDNA D-loop sequence and high genetic diversity,which may play an important role in the conservation and recovery of the species.

Abstract:The ultrastructure of mucosal epithelium of the digestive tract in Siganus canaliculatus was examined by scanning electron microscopy. In esophagus,longitudinal folds and V-shape secondary folds of mucosal surface were observed. There were numerous secretory holes and openings of glandular duct. The squamous epithelium cells were flattened,and they were firmly bound to one another. The stomach mucosal folds were arranged vertically,showing rope-like pattern. There were many papillae and no microvillus on the surface of stomach epithelium. The epithelium cells took the shape of pentagon or hexagon,and there were more gastric pits between them. Numerous villi arranged on the surface of intestinal epithelium,showing finger-like or flattened shape. The mucosal epithelium of intestine exhibited gyrus structure,and numerous secretory granules were on the surface. The epithelium cells of intestine were polygon,round or anomalous and had intercellular space. The epithelium structure of the pyloric caeca was similar to that in intestine. In the paper,the relationship between structure of epithelium and their functions was also discussed.

Abstract:Effects of water velocities (0,0. 1,0. 3 and 0. 5 m /s) on swimming performances of juvenile tinfoil barb Barbodes schwanenfeldi,weighing 125. 94 ± 13. 87 g,were investigated at the water temperature of 28℃ . Swimming behaviors of the fish were monitored continuously for 90 min by a video tape recorder,and the video data were analyzed by computer packages. The results indicated that rheotaxis frequencies and tail beat frequencies of juvenile tinfoil barb increased with increasing water velocities from 0 to 0. 3 m /s,but decreased at 0. 5 m /s. The time of swimming performances varied obviously at different water velocities. At 0 m /s,the fish kept moving for more than 98% of the experimental time,and at other three water velocities,the fish were in fixed positions against the current (FP) mainly. When the water velocities rose from 0. 1 m /s to 0. 5 m /s, the time of FP increased from 45. 8% to 81. 3% ,but that of moving forward against the current ( MF) decreased from 24. 1% to 5% or less. The time of moving backward against the current ( MB) reached the maximum at 0. 1 m /s,accounting for about 16. 4% of the experimental time. The time of downstream ( DS) accounted for 13. 7% ,2. 1% ,and 10. 9% of the experimental time at water velocities of 0. 1 m /s,0. 3 m /s and 0. 5 m /s,respectively. Tail beat frequencies of the fish showed significantly positive relationship with swimming speeds in the MF and FP performances,but had no significant correlation with swimming speeds in DS and MB performances.

Abstract:This paper describes the kidney microstructure of Rana nigromaculata. Histological and histochemical observation showed that besides nephrons and collecting tubes, lymphoid tissues could be observed in the kidney. Moreover,a structure that was similar to the corpuscle of Stannius of teleost was found at the ventral side of the kidney. Numerous mast cells could be observed in that structure as well,suggesting that the kidney of R. nigromaculata is a multifunctional organ.

Abstract:The hyobranchial apparatus features of adult and subadult Hynobius guabangshanensis were observed by clearing and double-staining method. In adult,the middle of hypohyal was crossed; the end of ceratohyal was covered by a complex of hyobranchial Ⅰ and ceratobranchial Ⅰ; basibranchial plate was present; ceratobranchial Ⅱ was ossified; urohyal was also ossified and showed a"一"shape. In subadult,the middle of hypohyal was not crossed; hyobranchial Ⅰand ceratobranchial Ⅰwere present separately; basibranchial plate was not presented,ceratobranchial Ⅱ was not ossified; ceratobranchial Ⅲ and ceratobranchial Ⅳ were present,and both were cartilage; urohyal was not ossified and showed a "1 " shape. These changes of morphological characters may be caused by the function of hynobranchial apparatus in adult and subadult.

Abstract:Characteristics of appearance,skull,scapula and fin bones morphology were described by 69 measurements taken from 14 specimens Sousa chinensis dead in the Pearl River Estuary. Those measurements were analyzed by one-way ANOVA to reveal the sexual dimorphism between individuals from different geographic locations. No difference was found in external measurements between male and female, but significant difference was detected in partial measurements on skull,scapula and fin bones between male and female. In general,the mean values of most measurements were larger in female than those in male.

Abstract:The aim of this study is to produce transgenic mice by testis-mediated gene transfer and investigate the suitable concentration of exogenous DNA for injection. A total of six SPF KM male mice were divided into three groups and liposome-treated exogenous DNA was directly injected into testis at three different concentrations:0. 08 μg /μl,0. 12 μg /μl and 0. 24 μg /μl,respectively. Every treated mouse was naturally mated with 3 female mice on the 5th day after the injection. The spermatozoa were detected on the 20th day after injection under the fluorescence microscope,and mouse testicular paraffin sections were prepared for detection of green fluorescent protein (GFP) expression under fluorescence microscope. The integration of pEGFP-N₁ in the F1 was determined by PCR. The results showed that the proportion of florescent sperm in 3 groups were 9. 09% , 47. 06% and 27. 78% ,respectively. Expression of GFP in different degrees was also found in three groups of paraffin sections. The PCR positive rates of offspring were 17. 26% ,47. 61% and 22. 11% ,respectively. It is concluded that the transgenic mouse could be produced by direct injection of foreign DNA into mouse testis,and 0. 12 μg /μl was the suitable concentration of exogenous DNA for injection in this study.

Abstract:Eight specimens of Paa robertingeri and one specimen of Odorrana hejiangensis were collected from Simian Mountain of Jiangjin District in Chongqing on 23 and 24 July,2009. These two species are first recorded in Chongqing.

Abstract:Tropidophorus hainanus( Scincidae) was collected in Hunan Mangshan National Nature Reserve on Aug. 4th,2009 and Amphiesma bitaeniata(Colubridae) was collected in Hunan Chengbu Mingzhulaoshan Nature Reserve on Aug. 10th,2009. Both of them are first recorded in Hunan Province.

Abstract:Molting in decapod crustaceans is coordinated by endocrine cues,and the primary axis of endocrine regulation comprises a steroid-producing epithelioid gland termed the Y-organ (YO) and a neurosecretory center termed the X-organ /sinus gland complex ( XO /SG). The signaling pathway activated by MIH,however, remains a controversy. This paper has summarized cellular mechanisms of the MIH regulation,including research methods and current three controversial issues: MIH receptors,functions of cAMP and cGMP,and functions of Ca²⁺ .

Abstract:With more than 100 years of history,studies on fish otolith have become a special field in the ichthyology,and fish age and growth in the studies is still one of the major themes. We reviewed the past research of otolith on fish age and growth from three aspects: otolith’s annual rings,daily increments and weight. Especially,the otolith daily increments’study has been and will continue to be the most important subject on the research of fish otolith. In our opinion,studies related to otolith will develop from individual to population gradually. Meantime,the sustainable use of fisheries resources will benefit a lot from the otolith study.