Catalogue of the Type Specimens of Amphibians and Reptiles in the Herpetological Museum of the Chengdu Institute of Biology,Chinese Academy of Sciences:Ⅱ.Hynobiidae (Amphibia,Urodela)

In order to facilitate herpetological research for herpetologists in and outside of China,we will report all the type specimens of Amphibia and Reptilia deposited in the Herpetological Museum of the Chengdu Institute of Biology(CIB),Chinese Academy of Sciences(CAS) in series. This report focuses on the type specimens of the family Hynobiidae of Urodela in Amphibia,which comprises seven species. Each entry provides the genus name,species name,information of the original description,the catalog number of holotype,sex,type locality,information of allotype(if any) and paratype,number and distribution of deposited specimens,and comments where they are appropriate.

Transmission of Carbapenem Resistance Between Human and Animal NDM-Positive Escherichia coli Strains

Although carbapenem use is prohibited in animals in China,carbapenem-resistant Escherichia coli(CREC),especially New Delhi metallo-β-lactamase(NDM)-producing strains,are widely prevalent in foodproducing animals.At present,the impact of livestock-associated CREC strains on human populations at the national level is unknown.Here,we conduct a retrospective cross-sectional study to investigate the prevalence of CREC from clinical settings across 22 Chinese provinces or municipalities and analyze anthropogenic factors associated with their presence.We also ascertain the blaNDMand blaKPCabundance among pig and chicken farms and present a detailed genomic framework for CREC of animal and human origin.Overall,631/29799(2.1%)clinical Escherichia coli(E.coli)isolates were identified as CREC.Multivariable analysis revealed that being male,an age below 1,an age between 13 and 18,provinces with greater chicken production,and provinces with higher pig production were associated with higher odds of CREC infection.In general,73.8%(n=45/61)of pig farms and 62.2%(n=28/45)of chicken farms had a blaNDMabundance of 1×10^(-5)to 1×10^(-3)and 1×10^(-3)to 1×10^(-2),respectively.Among all the Chinese NDM-positive E.coli(n=463)available at the National Center for Biotechnology Information(NCBI),the genomic analysis revealed that blaNDM-5and Inc X3 were the predominant carbapenemase gene-plasmid combination,while a highly homogeneous relationship between NDM-positive isolates from humans and animals was demonstrated at the plasmid and core genome levels.All the findings suggest frequent CREC transmission between humans and animals,indicating that further discussions on the use of antibiotics in animals and humans are needed,both in China and across the globe.

Lactic Acid Reduces LPS-Induced TNF-a and IL-6 mRNA Levels Through Decreasing I?Ba Phosphorylation

This study explored the effects over time of lactic acid （LA） on IκBα phosphorylation and nuclear factor-kappa B （NF-κB） p65 protein expression, and on tumor necrosis factor a （TNF-α） and interleukin-6 （IL-6） mRNA levels in rat intestinal mucosa microvascular endothelial cells （RIMMVECs） stimulated by lipopolysaccharide （LPS）. I？Ba, phosphorylated IκBa （p-IκBa） and p65 protein levels were monitored by Western blot analysis, and TNF-a and IL-6 mRNA levels were analyzed using real-time PCR. LA treatment reduced TNF-a and IL-6 mRNA levels in LPS-stimulated RIMMVECs, with the greatest effect being after 3 h. The highest inhibitory effect of LA on IκBa phosphorylation to prevent activation of NF-κB was after 6 h. These results suggest that LA reduces TNF-a and IL-6 mRNA levels through decreasing IκBa phosphorylation and blocking the dissociation of IKK complex, which prevents activation of NF-κB.

Relationships Between Icariin and Anti-Apoptotic miRNA-21 in Mouse Blastocyst Development In vitro

In this study, the effect of icariin, a flavonoid from the Chinese traditional medicine epimedium, on miRNA-21 of mouse developmental blastocysts in vitro and the development of preimplantation embryos were studied. The possible effective targets of icariin promoting preimplantation embryo development in vitro and anti-apoptosis were determined. The embryos were cultured in modified CZB medium （mCZB） as control group. The experimental group （Ica group） was supplemented with 0.6 μg mL-1 icariin. Mouse pronuclear embryos were cultured in vitro until blastocysts. The development rates of preimplantation embryos were observed. The total cell number, apoptotic cell number and the rate of apoptotic cells in blastocysts were analysed by the staining of Hoechst33342 and labeling of TUNEL and detected under a laser confocal scanning microscope. The miRNA-21 expression, the mRNA levels of pro-apoptotic Caspase3, and the target genes of miRNA-21： pro-apoptotic PTEN, anti-apoptotic Bcl-2 were detected by real-time RT-PCR. The results showed that percentages of morulaes and blastocysts in Ica group were both extremely higher than control group （（85.14±6.57）% vs. （72.04±11.58）%; （82.50±7.11）% vs. （66.80±11.70）%, respectively, P〈0.01）. The total cell number ofblastocysts had extreme difference between Ica group and control group （（61.40±9.64） vs. （46.23±4.50）, P〈0.01）. The apoptotic cell number and rate of apoptotic cells of blastocysts were both reduced in Ica group （（1.47±0.51） vs. （2.94±0.66）; （2.40±0.27）% vs. （6.25±0.62）%, respectively, P〈0.01）. Compared to control group, addition of icariin into mCZB extremely increased the expression of anti-apoptotic miRNA-21 （P〈0.01）, down-regulated pro-apoptotic Caspase3 （P〈0.05） and PTEN （P〈0.01）, up-regulated anti-apoptotic Bcl-2 （P〈0.01）. In conclusion, icariin could reduce the apoptosis, promote the embryo development in vitro by enhancing miRNA-21 expression to up-regulated anti-apoptotic genes and down-regulated pro- apoptotic genes. These apoptosis-related genes were regulated by miRNA-21.

Molecular cloning of the heat shock protein 20 gene from Paphia textile and its expression in response to heat shock

P. textile is an important aquaculture species in China and is mainly distributed in Fujian, Guangdong, and Guangxi Provinces. In this study, an HSP20 c DNA designated Pt HSP20 was cloned from P. textile. The full-length c DNA of Pt HSP20 is 1 090 bp long and contains a 5′ untranslated region(UTR) of 93 bp, a 3′ UTR of 475 bp, and an open reading frame(ORF) of 522 bp. The Pt HSP20 c DNA encodes 173 amino acid residues and has a molecular mass of 20.22 k Da and an isoelectric point of 6.2. Its predicted amino acid sequence shows that Pt HSP20 contains a typical α-crystallin domain(residues 77–171) and three polyadenylation signal-sequences at the C-terminus. According to an amino acid sequence alignment, Pt HSP20 shows moderate homology to other mollusk s HSPs. Pt HSP20 m RNA was present in all of the test tissues including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, with the highest concentration found in the gonad. Under the stress of high temperature, the expression of Pt HSP20 m RNA was down-regulated in all of the tissues except the adductor muscle and gonad.

The analysis of genetic diversity and differentiation of six Chinese cattle populations using microsatellite markers

A total of 321 individuals from six cattle populations of four species in a bovine subfamily in China were studied using 12 pairs of microsatellite markers. The genetic diversities within and between populations were calculated. The phylogenetic trees were constructed by （δμ）^2 and DA distances, and the divergence times between populations were estimated by （δμ）^2. Altogether, 144 microsatellite alleles were detected including 24 private alleles and nine shared alleles. Chinese Holstein had the largest number of private alleles （10）, whereas Bohai black and Buffalo had the smallest number of private alleles （2）. Chinese Holstein showed the highest genetic variability. Its observed number of alleles （Na）, mean effective number of alleles （MNA）, and mean heterozygosity （He） were 7.7500, 4.9722, and 0.7719, respectively, whereas, the Buffalo and Yak showed low genetic variability. In the phylogenetic trees, Luxi and Holstein grouped first, followed by Bohai and Minnan. Yak branched next and buffalo emerged as the most divergent population from other cattle populations. Luxi and Bohai were estimated to have diverged 0.039-0.105 million years ago （MYA）, however, buffalo and Holstein diverged 0.501-1.337 MYA. The divergence time of Yak versus Minnan, Holstein and buffalo was 0.136-0.363, 0.273-0.729, and 0.326-0.600 MYA, respectively.

Natural astaxanthin enhanced antioxidant capacity and improved semen quality through the MAPK/Nrf2 pathway in aging layer breeder roosters

Background:Natural astaxanthin(ASTA)has strong antioxidant properties and has been widely used as a health product to improve human health.However,the effects of ASTA on the reproductive performance of aging roosters have been poorly studied.We aimed to investigate the effects of dietary ASTA on semen quality and antioxidant capacity in aging roosters and to explore the potential mechanism of semen quality change via antioxidation defense system.Methods:In the present study,9653-week-old Jinghong No.1 layer breeder roosters were fed a corn-soybean meal basal diet containing 0,25,50,or 100 mg/kg ASTA for 6 weeks.Results:Semen quality in the ASTA groups remarkably improved than that in the control group,and antioxidant activities,the abilities to scavenge hydroxyl radicals and superoxide anions,increased gradually with ASTA addition(P<0.05).In addition,the mRNA levels of antioxidant enzymes as well as the mRNA and protein levels of the mitogen-activated protein kinase(MAPK)and nuclear factor-erythroid 2-related factor 2(Nrf2)were markedly increased in the 50-100 mg/kg ASTA group(P<0.05).Conclusions:Collectively,these results demonstrate that dietary ASTA may improve semen quality by increasing antioxidant enzyme activities and the ability to scavenge hydroxyl radicals,which may be related to upregulation of the MAPK/Nrf2 pathway.

Identification of Novel and Differentially Expressed MicroRNAs in the Ovaries of Laying and Non-Laying Ducks

MicroRNAs （miRNAs）, which post-transcriptionally regulate gene expression by binding to the 3′untranslated region of mRNAs to either inhibit or enhance translation, are involved in diverse biological processes. The use of high-throughput Solexa sequencing plays important roles in the discovery of miRNAs. In this study, we used high-throughput Solexa sequencing to identify novel duck miRNAs and compare the miRNA expression profiles in laying and non-laying duck ovaries. Using a bioinformatic analysis, we discovered 86 potential duck miRNAs similar to known chicken miRNAs and 43 unique sequences that matched known miRNAs of other species. We also found that miRNA variations and isoforms were widespread in our two RNA libraries, with most of the variations occurring in the 3′region of the miRNAs. Furthermore, we detected 55 miRNAs that exhibited significant expression differences between laying and non-laying ducks. Gene ontology （GO） and kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses of the potential targets of the differentially expressed miRNAs indicated these miRNAs may play key roles in the egg laying process. Finally, we confirmed the differential expression of 5 miRNAs in the laying and non-laying samples by qRT-PCR. Cumulatively, our work provides the first look at the miRNA expression profile of the duck ovary and provides novel insight into the roles of miRNAs in egg laying and reproduction.

Comparison of Skull Morphology in Two Species of Genus Liua(Amphibia:Urodela: Hynobiidae),L.shihi and L.tsinpaensis

Skull characteristics play an important role in the systematics of tailed salamanders, in this study, the skulls ofLiua shihi and L. tsinpaensis were compared using a clearing and double-staining technique. The results showed that in L. tsinpaensis, the vomerine tooth rows are in a ＂ ＂ shape, the length of the inner vomerine tooth series is nearly equal to that of the outer series, the vomerine tooth rows do not extend beyond the choanae, an ossified articular bone is absent, the basibranchial is rod shaped, the radial loops exhibit a figure-eight shape, the cornua has two cylindrical branches, the urohyal is rod shaped, and the end of the ceratohyal is not ossified; these features differ considerably from those of L. shihi. The ossification of the posterior portion of the ceratohyal and the present or absent of ossified articular might represent ecological adaptation to feeding in different environments.

Morphological Characteristics of the Dorsal Skin of Two Hynobiids and Their Adaptive Role in Aquatic and Terrestrial Habitats

The morphological characteristics of the dorsal skin of trunk in two species of hynobiid salamanders, Batrachuperus pinchonii and Hynobius chinensis were examined by light microscopy. The basic structures of the skin in the two species are similar and consist of two layers： epidermis and dermis. The epidermis consists of stratum corneum, stratum intermedium and stratum germinativum, while the dermis is composed of a stratum spongiosum and stratum compactum. However, some species-specific variation has been identified （e.g., the distribution of capillary vessels and gland cells, and the thickness of skin）. H. chinensis is a terrestrial species and only lives in water during breeding period, but B. pinchonii is aquatic and remains aquatic throughout its lifetime. The differences in the distribution of capillary vessels and gland cells are related to their different habitats, and show a morphological adaptation.

Baicalin Induces IFN-α/β and IFN-γ Expressions in Cultured Mouse Pulmonary Microvascular Endothelial Cells

We studied the effect of baicalin,an extract from Radix Scutellariae（a traditional Chinese medicine） in inducing mouse pulmonary microvascular endothelial cells（MPMVECs） to produce interferons（IFNs）.MPMVECs were cultured in vitro in the presence of different concentrations of baicalin（10,20,and 30 μg mL-1）,and the cells and the culture media were harvested at various time intervals.The proteins and mRNA levels（relative to β-actin） of IFN-α/β and IFN-γ were analyzed by RT-PCR and enzyme-linked immunosorbent assay（ELISA）.It was observed that baicalin substantially up-regulated the expression of IFN-α/β and IFN-γ.In all baicalin-treated groups,the relative levels of IFN-α/β and IFN-γ mRNAs peaked after 12 h of culturing,and IFN-α/β and IFN-γ proteins peaked after 24 h of culturing.These results suggest that baicalin can effectively induce the expression of IFNs in pulmonary microvascular endothelial cells,and thus potentially act as an antiviral compound.This study may provide background information for developing new antiviral drugs based on baicalin.

Characterization of the cDNA encoding a BPI/LBP homologue in venom gland of the hundred-pace snake Deinagkistrodon acutus

Bactericidal/permeability-increasing protein （BPI） and LPS-binding protein （LBP） play an important role in host defence. Current evidence shows that BPI/LBP may be widely existed in different cells and tissue types of animals. A full-length cDNA clone encoding a BPI/LBP homologue （dBPI）, 1757 bp in size, was characterized in venom gland of the hundred-pace snake Deinagkistrodon acutus. Its deduced amino acid sequence of 417 residues had 13.8% - 21.5% identity to BPI like 1 （BPIL1） and BPI like 3 （BPIL3） of other animals. Conserved cysteine residues which are involved in disulfide bond formation between the final strand of the N-terminal beta sheet and the long alpha helix of BPI are identified as Cys146-Cys183 of dBPI. Phylogenetic tree analysis showed that the BPI/LBP homologues formed five large clusters and dBPI was in a large cluster including BPIL1 and BPIL3. dBPI mRNA shows a tissue specific expression in venom gland. This is the first study to identify the cDNA encoding BPI/LBP homologues from reptiles [ Current Zoology 55 （5） ： 376 - 382, 2009].

Microsatellite-Based Genetic Differentiation and Phylogeny of Sheep Breeds in Mongolia Sheep Group of China

In the present study we studied the genetic structure of five Chinese sheep populations of Hu sheep, Tong sheep, Small-tailed Han sheep, Tan sheep, and Wadi （WD） sheep using 15 microsatellite loci. The results showed that the FIT, FST, and FIS statistics computed for the complete dataset had the following values, 0.523±0.140, 0.363±0.131 and 0.263±0.092, respectively. All loci were significantly contributed to the genetic differentiation among population （P0.001）. There is no relationship between the scatter of pairwise FST geographical distance points as geographical distance increases between the five populations. Membership probabilities and genetic structure of sheep populations were estimated when K=2, the populations were classified into Hu, Tong, Han and WD, and Tan sheep group. However, when K=3, the populations were classified into Hu and Tong, Han and WD, and Tan sheep group. The findings supported the previous literatures that these populations are originated on different time stages from the primogenitor population and communicated genetically with each other by natural and artificial selection in different ecological environment.

The Levels of Genetic Differentiation of Small-Tailed Han Sheep and Tan Sheep Populations Using Structural Loci

Using the method of ＂random sampling in typical colonies of the central area of the habitat＂ and several electrophoresis techniques, the variations of 17 structural loci encoding blood proteins in 60 Small-Tailed Han sheep and 73 Tan sheep were examined and compared with those of 14 other sheep populations in China and other countries to investigate their levels of genetic differentiation. The average heterozygosities of Small-Tailed Han sheep and Tan sheep were 0.2360 and 0.2587, respectively. The average polymorphic information content values were 0.1974 and 0.2102, respectively. The average effective numbers of alleles were 1.5723 and 1.5751, respectively. The coefficients of gene differentiation in the four groups （including 4, 6, 13, and 16 sheep populations, respectively） were 0.049323, 0.059987, 0.1728, and 0.201256, respectively, indicating that the degree of gene differentiation at the structural loci was the least in Hu sheep, Tong sheep, Small-Tailed Hart sheep, and Tan sheep; followed by the above-mentioned four sheep populations and two Mongolian sheep populations; and was the highest in sheep populations belonging to the Mongolian sheep group, South Asian sheep, and European sheep. The earlier researchers＇ conclusions that both Small-Tailed Han sheep and Tan sheep evolved from Mongolian sheep were further verified by the results of this study. Hu sheep, Tong sheep, Small-Tailed Han sheep, and Tan sheep were decreasingly affected by the bloodline of Mongolian sheep to different degrees. The relationships among sheep populations were not closely related to the geographical distances among sheep populations.

Determination of reference intervals for metabolic profile of Hanwoo cows at early, middle and late gestation periods

Background： Metabolic profile was initially designed as a presymptomatic diagnostic aid based on statistical analyses of blood metabolites to provide an early warning of certain types of metabolic disorder. However, there is little metabolic profile data available about Korean Hanwoo cows. Therefore, this study aimed to determine the reference intervals of metabolic profile for Korean Hanwoo cows. Methods： Healthy animals （2,205） were selected and divided into early （day 1 to 95）, middle （day 96 to 190） and late （day 191 to 285） period according to their gestating period. Metabolic profile including total protein （TP）, albumin （AIb）, urea （UREA）, glucose （Glu）, total cholesterol （T-Cho）, long-chain fatty acid （LCFA）, aspartate aminotransferase （AST）, gamma-glutamyl transpeptidase （GGT）, creatinine （Crea）, calcium （Ca）, inorganic phosphorous （iP） and magnesium （Mg） were analyzed using a TBA-4OFR automatic biochemical analyzer. The data of Korean Hanwoo cows were then compared to those of the Japanese Wagyu cows. Results： Most of the data of the Korean Hanwoo cows were relatively higher than those of Japanese Wagyu cows, with the exception of Glu and GGT. This may indicate that the nutritional level of feed for the Korean Hanwoo cows was higher than that of the Japanese Wagyu cows because of the different feeding system. In particular, relatively higher levels of UREA and LCFA were observed in the Korean Hanwoo cows, and this may also contribute to the low reproduction efficiency. Conclusions： These findings may provide some theoretical basis for understanding the reproductive and feeding situation of Korean Hanwoo cows.

Molecular cloning, characterization and expression analysis of a catalase gene in Paphia textile

Catalase is an important antioxidant protein that can protect organisms against various forms of oxidative damage by eliminating hydrogen peroxide. In this study, the catalase c DNA of Paphia textile（Pt CAT） was cloned using RTPCR and rapid amplification of c DNA ends（RACE）. Pt CAT is 1 921 bp long and consists of a 5′-UTR of 50 bp, a 3′-UTR of 349 bp, and an ORF of 1 542 bp that encodes 513 amino acids with a molecular weight of 58.4 k D and an estimated isoelectric point of 8.2. Sequence alignment indicated that Pt CAT contained a highly conserved catalytic signature motif（^（61）FNRERIPERVVHAKGAG^（77））, a proximal heme-ligand signature sequence（^（352）RLFSYSDP^（359））, and three catalytic amino acid residues（H^（72）, N^（145）, and Y^（356））. Pt CAT also contains two putative N-glycosylation sites（^（34）NKT^（36） and ^（437）NFT^（439）） and a peroxisome-targeting signal（^（511）AQL^（513））. Furthermore, Pt CAT shares 53%–88% identity and 29%–89% similarity with other catalase amino acid sequences. Pt CAT m RNA was present in all tested organs, including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, but its expression was highest in the digestive gland. High-temperature-induced stress produced two expression patterns of Pt CAT m RNA： first, an initial up-regulation followed by a down-regulation in the heart, digestive gland, and gonad and, second, consistent down-regulation in all other organs. These results demonstrate that Pt CAT is a typical member of the catalase family and might be involved in the responses to harmful environmental factors.

Effects of Bcl-2 Gene Interference on the Apoptosis,Proliferation and Progesterone Secretion of Goose Follicular Granulosa Cells

Based on published sequences for chicken Bcl-2,three siRNAs（small interfering RNA）were designed,and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro.Bcl-2 protein,apoptosis and proliferation of granulosa cells,48 h after the transf ection,were analyzed by flow cytometry,and progesterone（P）secreted into the culture medium was measured by radioimmunoassay.In addition,apoptosis and Bcl-2 protein level were assessed in untreated granulosa cells from the four largest preovulatory follicles（F1F4）,the smallest preovulatory follicles（SPF）,small yellow follicles（SYF）and atretic follicles.The highest level of Bcl-2 protein was observed in granulosa cells from SPF,and levels in cells from healthy follicles were significantly higher than those of atretic follicles（P【0.05）.Bcl-2 protein levels in cells subjected to RNAi were significantly lower than those of controls（P【0.05）,while apoptosis indices（AI）,proliferation indices（PI）and P secretion in the RNAi treatments were higher than those of controls（P【0.05）.

Analysis of novel immune-related genes and microsatellite markers in the transcriptome of Paphia undulata

Increasingly,exogenous stressors such as pathogen infections,variable water conditions,and pollution are resulting in high mortality of Paphia undulata,deleteriously affecting the quality of clam harvests.The foot is a burrowing organ in clams.Physical damage and constant contact with the external environment cause the foot to be highly sensitive to pathogen invasion and water condition variation.In the present study,the foot tissue transcriptome was analyzed to identify genes involved in immune and stress responses.The P.undulata transcriptome included 5 286 668 078 bp reads generated by Illumina Hiseq 2000 sequencing and were assembled into 1 785 226 contigs by de novo method.The contigs were clustered into 99 339 transcripts and further grouped into 60 201 unigenes.Of them,22 260 unigenes were successfully annotated using public databases.Twelve genes that were response to immune and stress were identified with abundant expression levels,including heat shock protein 70,cold shock protein,complement C3,cathepsin L,ubiquitin carboxyl-terminal hydrolase L5,and translationally controlled tumor protein.Furthermore,566 unigenes were found homologous to genes involved in the immune response systems of pathogen discrimination,signal transduction,and immune effector,such as lectins,toll-like receptors,complement pathway,toll-like receptor signaling pathway,heat shock proteins,antioxidant enzymes,lysozymes,and mucins,indicating that P.undulata could have a complete set of innate immune mechanisms.In addition,4 270 microsatellite markers(SSRs)were identified from 60 201 unigenes,of which trinucleotide repeats were most abundant and 16 SSRs were tested to be polymorphic.The present study provides a new insight into innate immunity and stress response mechanisms in P.undulata.

Heat,sight and scent:multiple cues influence foraging site selection by an ambush-foraging snake Hoplocephalus bungaroides(Elapidae)

Most mobile organisms respond to multiple cues when selecting habitat types,and laboratory experiments that manipulate only single cues may fail to reveal the true complexity of habitat-selection behaviour.In south-eastern Australia,broad-headed snakes Hoplocephalus bungaroides(Elapidae)lie in wait under sun-warmed rocks to ambush velvet geckos Oedura leseuerii(Gekkonidae).Previous laboratory work has shown that both the geckos and the snakes actively select hotter rather than colder rocks,and that the snakes actively select rocks scented by geckos.We manipulated rock temperature and the presence of two types of cues from geckos(chemical and visual information)to clarify the causal basis for foraging site selection by the juveniles of this snake.When given a choice between cold lizard-scented rocks and hot unscented rocks,our captive snakes gave a higher priority to lizard scent than to temperature.The snakes also selected shelter-sites that provided visual as well as scent cues from lizards,rather than shelter-sites with scent cues alone.Thus,although broad-headed snakes show a direct preference for hotter rather than colder rocks in the laboratory,their choice of foraging site in the field may also be influenced by the presence of scent cues from prey.Our laboratory results suggest that habitat selection by broad-headed snakes may be more complex than has been suggested by previous single-factor laboratory trials.

The Genetic Diversity and Phylogenetic Status of Luxi Cattle

A total of 87 individuals of Luxi cattle from Juanchen and Liangshan counties, Shangdong Province, China, were sampled by simple random sampling in typical colony. Twenty-one blood proteins and enzymes loci were detected by polyacrylamide gel electrophoresis （PAGE） and starch gel electrophoresis （SGE）. In the meantime, the data of 7 loci of 13 cattle populations in China and other countries were collected and phylogeny relationships were studied. The results indicated that 9 out of 21 loci showed polymorphism （42.86%）; the level of genetic variation in Luxi cattle population was relatively high, the mean heterozygosity was 0.1416. The Luxi cattle have a close phylogenetic relationship with the cattle populations of east and south of Asia, and this further confirmed the fact that Luxi cattle were the cross-breed between the Bos taurus and Bos indicus in China, but it is impossible that yellow cattle contained the blood of of Bali.