2008 Vol. 29, No. 6

Display Method:
The nuclear insertions of mitochondrial DNA (Numts), which originate from the integration of nuclear DNA by mtDNA, are found as molecular fossils in the nuclear genomes of various eukaryotes. Because integrated Numts tend to have a high sequence similarity to genuine organellar mtDNA sequences, inadvertent amplification of Numts can be a nuisance in studies of mtDNA variation. With the availability of the complete domestic horse genome sequence, we present the first comprehensive analysis of genome-wide distribution and frequency of Numts in the nuclear genome of domestic horse (Equus caballus). In the present paper, we detected 200 Numts ranging between 29 and 3 727 bp in size, which collectively representing only 0.002154% of the nuclear genome. Furthermore, ten of these segments were found to be longer than 800 bp. The absence of Numts in mitochondrial control region suggested that it would not influence the analysis of horse population genetics studies relating to this region. We also found that during horse evolution, Chromosomes 1 and 27 have been more susceptible to integration by Numts. The results in this study may provide valuable information for future mtDNA studies in Equidae species, including its use as a tool for avoiding Numt contaminations that may result in inauthentic results of experimentation.
The phylogenetic relationships among all taxa within the Acrididae (Orthoptera: Acridoidea) were largely unknown until now. In this study, to further investigations, 24 species of Acrididae from China were used as sample taxa. The sequence constitutions and variations were analyzed and the molecular phylogenetic trees were reconstructed based on the combined sequence data (795 bp length in total) of 12S rDNA and 16S rDNA, using the grasshopper Pyrgomorpha conica of Pyrgomorphidae as the outgroup. The results showed that the rates of the two kinds of transitions are obviously much higher than that of the four kinds of transversions in these combined 12S+16S rDNA sequence data. The saturation of nucleotide substitutions happened in 12S and 16S rDNA sequence data. The molecular phylogenetic trees indicated that Oedipodinae is a monophyletic group and this subfamily is a natural one, but Catantopinae and Acridinae are non-monophyletic. Oedipodnae is a relatively primitive group within the Acrididae, whereas the Oxyinae may have diverged later than Oedipodinae, but earlier than most other species of Acridide.
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.
The goal of this study was to investigate the genetic stability of the C57BL/6J (B6) inbred mouse strain maintained in different breeders. Three populations of B6, Pop1 and Pop2 purchased from Beijing and Pop3 purchased from Shanghai, were examined. Fifteen microsatellite loci reported to be polymorphic among inbred strains were amplified using FAM labeled primers and genotyped with ABI Prism 377 automated sequencer. Seven loci were found polymorphic, and all the loci were homozygous in all the three populations. The present study indicates that genetic variation occurs in different B6 populations although they are still inbred in each breeder. The mechanism of genetic variation is not well understood now, but it is very important to know the precise content of the B6 genome before use of this strain in research.
Partial cDNA sequence of rabbit BMP15 was cloned by RT-PCR from rabbit ovaries, showing a similarity of 83%−90% with the BMP15 nucleotide sequences in humans, mice, ovine, sheep, cows and pigs. The expression of BMP15 in rabbit cumulus-oocyte complexs during oocytes in vitro maturation (IVM) was measured by fluorescent quantitative RT-PCR method. BMP15 was expressed at low levels in immature oocytes and increased to the highest level at 16 h of IVM, which coincides with the time of cumulus cell expansion, then declined slowly under IVM cultivation. The expression pattern of BMP15 suggested that it might be important in cumulus expansion in rabbits.
Mice were administrated by the consecutive morphine, subcutaneous injection (25, 50, 75, 100, 125, 150 mg/kg)to establish the withdrawal model. The naloxone (6 mg/kg)-induced responses, namely jumping behavior can be observed as withdrawal behavior. Bromocriptine and baclofen were used in the protocol and their synergetic inhibition was also studied. The results suggested that the low dose of bromocriptine (10 mg/kg) can not inhibit the withdrawal syndromes , whereas the mid (20 mg/kg) and high (30 mg/kg) doses can do. The low (0.5 mg/kg) and mid (1.0 mg/kg) doses of baclofen can not decrease the withdrawal syndromes, whereas the high dose can effectively inhibit the nalxone-induced response in the mice. However the co-administration of bromocriptine (10 mg/kg) and baclofen (1.0 mg/kg) can inhibit the withdrawal response. The results showed that the two receptors synergized and effectively inhibited the naloxone-induced withdrawal syndromes.
The interaction of morphine and cholinergic system was shown in previous studies. In the present study, we investigated whether morphine would interact with the cholinergic antagonists, scopolamine and atropine in a Y-maze spatial recognition memory. Pre-test treatments of morphine (5, 1.5, 0.5 mg/kg), scopolamine (1, 0.1 mg/kg), atropine (0.5, 0.1 mg/kg) were used in the experiments, relatively high or low doses were paired respectively as co-administration measures. The results showed that co-administration of morphine 0.5 mg/kg + scopolamine 0.1 mg/kg and morphine 0.5 mg/kg + atropine 0.1 mg/kg disturbed the inspective exploratory behavior (percent of arm duration) but not the inquisitive behavior (percent of arm visits) of the spatial memory retrieval, while the drugs didn’t cause amnesia when single administered of the concerned low doses. Distinct interaction was found between scopolamine and morphine on increasing locomotor activity.
The mice with a continuous 7-day intraperitoneal injection of morphine was induced the conditioned placed reference (CPP) model and the mice with a continuous 7-day subcutaneous injection of morphine(25, 50, 75, 100, 125, 150 mg/kg)was induced withdrawal model (jump behavior) by naloxone administration(6 mg/kg). The results suggested that the administration of the GABAB receptor agonist, baclofen, could effectively attenuate the expression of CPP and the nalxone-induced response in the mice. It could be concluded that activating GABAB receptor can alleviate physical and psychic dependence syndromes to a certian extent .
Antimicrobial proteins and peptides had been found from a wide variety of organisms in the last few years. These molecules have attracted much research interest because of their biochemical diversity, broad specificity on anti-viral, anti-bacterial, anti-fungi, anti-protozoan parasites, anti-tumoural, and wound-healing effects. Antimicrobial proteins and peptides play key roles in innate immunity. They interact directly with bacteria and kill them. The brown-spotted grouper, Epinephelus fario, is an important marine fish cultured in southern China. Recently, bacteria and virus have caused high mortality in E. fario cultures, but its endogenous antimicrobial peptides and proteins have not been explored. An antimicrobial component was found from the skin homogenate of E. fario. After the skin homogenate was digested with trypsin, its antimicrobial activity was lost, which showed that the antimicrobial component is a protein. The antimicrobial protein (Efap) was purified from the skin homogenate of E. fario by successive ion-exchange and gel filtration chromatography. Efap was demonstrated to be single protein band by SDS-PAGE, with the apparent molecular weight of 41 kD. Efap exhibited antimicrobial activity both for the Gram-positive bacteria, Staphylococcus aureus, Micrococcus luteus and Bacillus subtilis, and for the Gram-negative bacteria, Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio fluvialis, Pasteurella multocida, Aeromonas hydrophila, Eschrrichiu coli, and Pseudomonas aeruginosa. Except A. hydrophila, P. aeruginosa, and E. coli (MIC>20 mol/L), most of the tested Gram-negative bacteria were sensitive to Efap (MIC<20 mol/L). Interestingly, Efap showed potent antimicrobial activity against Gram-positive bacteria S. aureus (MIC 5-10 mol/L) but comparatively weak antimicrobial activity against M. luteus and B. subtilis. The broad antimicrobial activities of Efap suggest that it contributes to the innate host defence of E. fario.
The aim of this study was to establish a method of isolating and culturing smooth muscle cells from the ductus deferens of rats. Smooth muscle cells were prepared from ductus deferens by explanting technique after dissection of adventitia and intimae, and cultured in vitro. The identification of the smooth muscle cells were verified by using anti α-smooth muscle actin (α-SMA) immunohistochemistry studies. The result suggested that the cells are multi-morphous, showing long fusiform or star shapes. The apophysis of cells contacted and coalesced to each other, in some regions the cells overlapped in multilayer, while in the other regions they formed monolayer that fluctuated and showed a “peak-valley” shape. They presented a positive reaction through immunohistochemistry studies. The purity of the cells was more than 99% through this method. The culturing of smooth muscle cells by explanting technique is simple and stable.
Five years’ (2000–2004) continuous study has been carried out on small mammals such as rodents in seven different sample plots, at three different altitudes and in six different ecological environment types in the eastern part of the Wuling Mountains, south bank of the Three Gorges of Yangtze River in Hubei. A total of 29 297 rat clamps/times were placed and 2 271 small mammals such as rodents were captured, and 26 small mammals were captured by other means. All the small mammals captured belonged to 8 families 19 genera and 24 species, of which rodentia accounted for 70.83% and insectivora 29.17%. Through analysis of the data, the results showed that: 1)although the species richness had a trend of increasing along different sample plots as altitude increased from south to north, quite a few species showed a wide habitat range in a vertical distribution(15 species were dispersed over three zones and two species over two zones), indicating a strong adaptability of small mammals such as rodents at lower altitudes in most areas and comparatively less vertical span of entire mountains; 2)whether in seven different sample plots or six different ecological types, Apodemus agrarius and Rattus norvegicus were dominant species below 1 200 m, and Anourosorex squamipes, Niviventer confucianus and Apodemus draco were dominant above altitudes of 1 300 m,however, in quantity they were short of identical regularity,meaning they did not increase as the altitude did,or decrease as the ecological areas changed; 3)the density in winter was obviously greater than that in spring, and the distribution showed an increasing trend along with altitude, but the density in different sample plots was short of identical regularity, showing changes in different seasons and altitude grades had an important impact on small mammals such as rodents; 4) in species diversity and evenness index, there were obvious changes between the seven different sample plots, probably caused by frequent human interference in this area. Comparatively speaking, there was less human interference at high altitudes where vegetation was rich and had a high diversity and evenness index, and the boundary effect and community stability were obvious. Most ecological types have been seriously interfered with due to excessive assart at low altitudes with singular vegetation and low diversity and evenness index and poor community stability, showing an ecosystem with poor anti-reversion. If human interference can be reduced in those communities at high altitudes with low diversity and evenness index, the biological diversity in the communities will gradually recover to similar levels of other ecological areas.
Plant communities of foraging sites of Hume’s Pheasant (Syramticus humiae) had been surveyed by systematic sampling method since April 25 to May 16, 2008 in Nanhua part of Ailaoshan National Nature Reserve. And 133 plant species were recorded, belonging to 86 genera and 49 families. The results by spearman correlation analysis showed that there was a significant relativity between occurrence frequency of S. humiae and distribution of evergreen broadleaf forest, which meant the vertical distribution and territory of the birds were affected by the distribution of evergreen broadleaf forest. S. humiae preferred to select evergreen broadleaf forest as habitats. The results by important value ordination of plant and detrended correspondence analysis showed that the plant composition in foraging sites was conformed to evergreen broadleaf forest whereas greatly different from other forest types. The composition among plant layers in evergreen broadleaf forest could provide preferable coverage, either leaves or stalks of pteridophyte and nuts of fagaceae were foods for S. humiae. The results of diversity comparison showed that plant diversity in foraging sites were significantly higher than Pinus armandii forest and deciduous broadleaf forest because both forests lacked vegetable foods. So habitat selection of Hume’s pheasant was affected by plant diversity and food richness. The results by comparison of vegetation factors and detrended correspondence analysis showed that tree coverage in foraging sites were higher than other forest types except evergreen broadleaf forest. The degree of similarity in foraging sites and other forest types differed. Deciduous broadleaf forest was less similar to foraging sites in vegetation factors where trees were sparse and S. humiae liked to graze greatly. There were short trees and few shrubs in P. armandii forests. However, mixed broadleaf and coniferous forest were more similar to foraging sites in vegetation factors, where human disturbance was serious. Coverage and human disturbance were dominant factors that influenced foraging sites selection of S. humia. The results by analysis of dissimilarity showed that the most studied area could supply basic conditions for the survival of S. humiae but the best suitable area was less indeed.

Black-and-white snub-nosed monkeys (Rhinopithecus bieti) distribute in a restricted area of the TransHimalayas between the Mekong and Yangtze River, at 26PoP14’N−29PoP20’N and 99PoP15’E−99PoP37’E. There are about 1 700 individuals in 15 groups remained in the habitat between 4 200 (north) −2 600 m (south) asl. The Milaka group is the northernmost range of the species with about 50 individuals in Mangkang county of Tibet. Based on our field survey and previous reports, we identified the fir forest and the mixed conifer forest as suitable habitat for the monkeys. Summer grazing lands and farmlands, which were made by local people’s cutting and burning in the fir forest at the high and low altitude belt, are replacing fir forest. To evaluate the status of the monkeys’ habitat, we employed GIS and RS to identify the habitat types with Landsat TM satellite imagery in winter of 1986 and 2006 respectively. The work resulted in: 1) the size of summer grazing lands, farmlands, and fir forest was 4 900 hmP2P, 3 300 hmP2P and 13 600 hmP2P in 2006 respectively; 2) during the past 20 years (1986−2006), the size of fir forest decreased by 15.5% (2 500 hmP2P), summer grazing lands and farmlands increased by 58.1% (1 800 hmP2P) and 17.8% (500 hmP2P) respectively; 3) the habitat of the species was more fragmented, the number of habitat patches increased by 75.6%, the mean size of forest patches decreased by 51.8% (from 15.3 to 7.4 hmP2P), the largest patch index decreased by 54.7%; the patch richness remained the same, but the Shannon’s diversity index and the Shannon’s evenness index increased by 4.0%, respectively; and 4) the size of fir forest negatively correlated with villager population (r =−1.000), but the size of summer grazing lands and farmlands positively correlated with villager population (r = 1.000). These indicate the habitat lost and fragmentation for the Milaka group increased sharply during the past 20 years and it is the result of population growth and the most employment of traditional modes.

The color preference of the Parasite Wasps of Ericerus pela was studied in the white wax garden. Eight kinds of color boards (red,yellow,blue,blue-green,green,grey brown,white and black) were used to attract the parasite wasps of Ericerus pela. The active regularity of the parasite wasps E. pela were observed during four periods per day ( 6:00-9:00,9:00-12:00,12:00-
The Hainan gibbon (Nomascus hainanus) is one of the most endangered primates and the least studied gibbon species in the fieldwork. In the past decades only a little information was known about its population size, population structure, habitat and food kinds. Through four years field work, we observed their inter-group encounters for four times. We found that their encounter behaviors were much different from other gibbon species. Only vocal and chasing behavior happened between the adult females, male sub-adults and male juveniles of the Hainan gibbon encounters. The time of inter-group encounters was much shorter than other gibbon species, just 24-51 minutes. Not like the Hylobates lar encounters, no fighting was observed among the Hainan gibbon encounters; there was also no such behaviors as playing and grooming, even the extra-group-copulations. The adult females of two groups didn’t take par in the chasing and they only rested and watched 20-30m away. We speculated that the behavior of the adult females may be interpreted as protecting their territory and by the way of chasing the sub-adults can learn how to protect their territory in the future.
A new species of the genus Triplophysa was discovered in underground water of a cave near Nijiao Village, Qiubei County (24°05′N,104°01′E), Yunnan, China, in 2002 and 2006. The new species, Triplophysa qiubeiensis is similar to T. anterodorsalis Cao et Zhu (1989), and can be distinguished from all the cave-dwelling Triplophysa in China by the following characters: 1) eyes highly degenerated vs. eyes normal; 2) branched dorsal–fin ray 7 vs. 8-9; 3) caudal fin forked vs. emarginate. The new species is also similar to T. macromaculeta Yang (1990) from the same river system, Nanpan Jiang, but can be distinguished from the latter the following characters: 1) eyes highly degenerated vs. eyes normal; 2) without blotches vs. with blotches; 3) caudal fin forked vs. emarginate.
In vertebrates, βγ-crystallins are mainly expressed in the lens as structural protein, a lot of non-lens βγ-crystallins were also discovered in microbes and mammals, such as Protein S and Absent in Melanoma 1, which act as microbial stress-inducible protective protein in bacterial, participate in epidermis development and has tumor suppression function in mammals, respectively. However, little is known about the biochemical properties, functions and action mechanisms of these non-lens βγ-crystallins in vertebrates. In this paper, we mainly review the research progression of non-lens βγ-crystallins in the protein structure, gene evolution and biological function.