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Geographical features and palaeoclimatic fluctuations are two classical evolutionary forces that shape genetic diversification within species. Fine-grained analysis of the mechanisms involved through population demographic processes, however, remains limited. Taking advantage of two recently published reference genomes, we resequenced the genomes and examined the evolutionary history of the moustache toads, a group endemic to East Asia where complex topography and fluctuating palaeoclimate are known to have had profound impacts on organisms. Moustache toads probably originated in southeast Yunnan, China, and diversified towards the northwestern of Yunnan, as well as central and eastern China. Further exploration based on three widespread species (Leptobrachium ailaonicum, L. boringii and L. liui) using demographic modelling and species distribution models revealed that mountains and river valleys in East Asia not only functioned as geographical barriers, but also provided dispersal corridors and facilitated continuous migration or post-glacial secondary contact among moustache toad populations. Furthermore, periodic oscillation of effective population sizes accompanying fluctuations of historical temperature and population contraction at the Last Glacial Maximum support the widespread impact of climatic changes of the Pleistocene on species diversification in East Asia. This impact was moderate for populations of L. ailaonicum and L. boringii in the southwestern mountains but severe for populations of L. liui in the eastern lowland regions of continental East Asia, which is supported by different degrees of change of their effective population sizes. Our findings reveal mechanisms underlying genetic diversification among moustache toads, and highlight the power of genomic data and demographic modelling for examining complex historical population-level processes and for understanding how geographical and palaeoclimatic factors interactively shape current intraspecific diversity.

To explore the synergistic immunomodulatory mechanism of interferon alpha-1b, interleukin-2 and thalidomide (ITI) regimen on patients with acute myeloid leukemia (AML).Sixty eight untreated de novo or relapsed or refractory or maintenance therapy patients with AML admitted in the Affiliated Cancer Hospital of Zhengzhou University and the other 11 medical units from March 2016 to May 2019 were treated with ITI regimen. Peripheral blood specimen per patient was collected into EDTA-K3 anticoagulation vacuum tube before the administration of ITI and 3 months after the treatment; peripheral blood lymphocyte subsets and perforin and Granzyme B expression were analyzed by using flow cytometry; the levels of VEGF, IFN-γ, TNF-α and IL-6 in the plasma were detected by using a cytometric bead array. Thirty-five healthy subjects from the hospital physical examination centre were selected as normal controls.The ratio of CD4The ITI regimen can raise the ratio of CD4干扰素α-1b、白介素-2联合沙利度胺方案治疗急性髓系白血病免疫调节作用的协同机制研究.探讨干扰素α-1b、白介素-2联合沙利度胺（简称"干白沙"方案）治疗急性髓系白血病（AML）的免疫调节的协同作用机制。.对2016年3月至2019年5月郑州大学附属肿瘤医院等11家医疗单位收治的初治或复发/难治性或行维持治疗的且自愿接受"干白沙"方案治疗的68例AML患者，采集患者用药前1天及规范用药3个月后的晨起空腹外周血标本，采用流式细胞术检测外周血淋巴细胞亚群及NK细胞颗粒酶B及穿孔素表达水平；微量样本多重定量技术（CBA）检测血浆中VEGF、IFN-γ、TNF-α、IL-6等4种细胞因子的表达水平。对照组为35例本院体检中心的健康体检者。.治疗前患者外周血CD4"干白沙"方案可提高外周血CD4

Genes encoding the major histocompatibility complex (MHC) are excellent candidates for elucidating adaptive variation because of their essential role in immune function. Hypotheses for how genetic variations of MHC genes are maintained include balancing selection, diversifying selection and neutral events; however, which of these forces are dominant remains controversial. In this study, we determined the preliminary forces that shaped MHC geographical variation in Omei tree frog by comparing with neutral microsatellites. The results revealed high genetic diversity and significant population differentiation in MHC genes of this species. The differentiation patterns of MHC and microsatellites were uncorrelated. Stronger level of genetic differentiation in MHC genes compared with that in microsatellites (based on comparison in both genetic differentiation patterns and strength of isolation by distance patterns) indicates the effects of diversifying selection on the geographical variation of MHC genes. Considering the significant evidence of positive selection acting on antigen-binding sites, we presumed that selection pressures may be mainly from locally different pathogens when gene flow is restricted. The results of this study contribute to a more comprehensive understanding of the evolutionary forces that maintain MHC geographical variation among natural populations.

Understanding habitat quality and landscape connectivity and exploring corridors connecting habitat patches are crucial for conservation, particularly for species distributed among isolated populations. The Sichuan golden snub-nosed monkey, Rhinopithecus roxellana, is an Endangered primate species endemic to mountainous forests in China. Its easternmost distribution lies in the Shennongjia area, which harbors an isolated subspecies, R. roxellana hubeiensis. Unfortunately, it has experienced significant habitat loss, fragmentation, and dramatic population decline in recent decades, primarily due to increased human disturbance. To quantify habitat quality, identify suitable habitat patches, and detect possible linkages among these patches for R. roxellana hubeiensis, we conducted habitat suitability assessments and landscape connectivity analyses in the Shennongjia area based on a set of environmental factors. We created a habitat quality model and a movement cost surface for the Shennongjia area based on a habitat suitability index, graph theory, expert knowledge, field experience, and information from the literature. Our results show that suitable habitat for R. roxellana hubeiensis in Shennongjia is fragmented and limited, and that this is particularly true for highly suitable habitats. We detected six core habitat patches and six least-cost paths and corridors. Our study does not provide accurate distributions of the monkeys and their habitat use. However, it identifies the most feasible and traversable habitats and corridors, which should be conservation priorities for this subspecies, and provides valuable guidance for reevaluating habitat conservation plans.

Man-Xiu Li, Li-Ting Jin, Tie-Jun Wang, Yao-Jun Feng, Cui-Ping Pan, Dei-Mian Zhao, Jun Shao Department of Breast Cancer, Hubei Cancer Hospital, Wuhan, People’s Republic of China Background: Triple-negative breast cancer (TNBC) is a subtype of breast cancer with poor clinical outcome and limited treatment options. Lacking molecular targets, chemotherapy is the main adjuvant treatment for TNBC patients. Materials and methods: To explore potential therapeutic targets for TNBC, we analyzed three microarray datasets (GSE38959, GSE45827, and GSE65194) derived from the Gene Expression Omnibus (GEO) database. The GEO2R tool was used to screen out differentially expressed genes (DEGs) between TNBC and normal tissue. Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed using the Database for Annotation, Visualization and Integrated Discovery to identify the pathways and functional annotation of DEGs. Protein–protein interaction of these DEGs was analyzed based on the Search Tool for the Retrieval of Interacting Genes database and visualized by Cytoscape software. In addition, we used the online Kaplan–Meier plotter survival analysis tool to evaluate the prognostic value of hub genes expression in breast cancer patients. Results: A total of 278 upregulated DEGs and 173 downregulated DEGs were identified. Among them, ten hub genes with a high degree of connectivity were picked out. Overexpression of these hub genes was associated with unfavorable prognosis of breast cancer, especially, CCNB1 overexpression was observed and indicated poor outcome of TNBC. Conclusion: Our study suggests that CCNB1 was overexpressed in TNBC compared with normal breast tissue, and overexpression of CCNB1 was an unfavorable prognostic factor of TNBC patients. Further study is needed to explore the value of CCNB1 in the treatment of TNBC. Keywords: triple-negative breast cancer, hub genes, expression profiling data, CCNB1

The promotion of angiogenesis is a promising therapeutic strategy for ischemic stroke. Many long noncoding RNAs (lncRNAs) are related to angiogenesis following ischemic stroke. LncRNA small nucleolar RNA host gene 12 (SNHG12) was upregulated in oxygen-glucose deprivation (OGD)-exposed primary brain microvascular endothelial cells and in microvessel from middle cerebral artery occlusion (MCAO) animal brains. However, the role and underlying mechanism of SNHG12 in ischemic stroke especially associated with angiogenesis process remain unknown. The expression of SNHG12 and miR-150 was determined in OGD-stimulated mouse brain microvascular endothelial (bEnd.3) cells. The role and mechanism of SNHG12 in the angiogenesis after ischemic stroke were investigated using gain- and loss-of function approaches both in OGD-exposed bEnd.3 cells and in MCAO mouse models. We found SNHG12 expression was elevated, whereas miR-150 reduced in OGD-exposed bEnd.3 cells. Upregulation of SNHG12 elevated, and SNHG12 knockdown suppressed the capillary-like tube formation, viability, migration, and VEGF expression in OGD-injured bEnd.3 cells. miR-150 mimic reversed, whereas anti-miR-150 further strengthened the effect of SNHG12 upregulation on the angiogenesis in bEnd.3 cells. Furthermore, we found that SNHG12 functioned as a competing endogenous RNA for miR-150 to regulate VEGF expression. Additionally, overexpression of SNHG12 improved the recovery of neurological function, reduced infarct volume and miR-150 expression, increased vascular density and VEGF expression in the infarct border zone of MCAO mice. In conclusion, SNHG12 promotes the angiogenesis following ischemic stroke via miR-150/VEGF pathway, which further clarified the mechanism of angiogenesis after ischemic stroke and provides a target for the treatment of this disease.

To investigate the application value of immunohistochemistry detection and bone marrow morphology analysis in the diagnosis and clinical staging of non-Hodgkin's lymphoma.Sixty-four cases of non-Hodgkin's lymphoma were selected in our hospital and the immunohistochemistry detection of pathologic specimens was perforemed by related monoclonal antibody and the bone marrow morphology was also examined.The positive rate of antibody CD79a on B cell lymphoma was 84.62%, and CD20 was 91.43%; the positive rate of antibody CD45RO on T cell lymphoma was 87.50%, and 88.89% of CD3. The bone marrow involvement was found in 10 cases (15.63%), 8 cases(12.50%) progressed to lymphoma - leukemia period; lymphoma cells were lower than 5% in 20 cases (31.25%).Immunohistochemistry detection can help to define the type of T or B cell lymphoma in patients with non-Hodgkin's lymphoma, the bone marrow morphology analysis can clear whether the patients with non-Hodgkin's lymphoma suffered from bone marrow involvement and whether developed to lymphoma- leukemia period. These methods possess more high value in clinical application.

Male-male competition and female mate choice based on ornamentation and genetic traits are the main drivers of animal sexual selection and group spawning. If male-male competition is intense, males with specific phenotypes should have advantages in breeding success or occupying superior mating positions. If female choice is important, females should have preferences for mate relatedness or males with good genes or optimal genetic compatibility against themselves. To detect the intensities of male-male competition and female choice and test the good genes and genetic compatibility hypotheses, we observed breeding behaviors, measured individual body lengths and breeding success indicators, and calculated male genetic heterozygosity, male–female relatedness, and genetic dissimilarity in an Omei treefrog (Rhacophorus omeimontis) population in Badagongshan, China. Our analyses showed that larger males obtained larger mates, had more mating opportunities, occupied better amplectant positions, and produced more offspring. However, females showed no inbreeding/outbreeding bias in mate choice, and the good genes and genetic compatibility hypotheses were not supported in female selection on mates and amplectant positions. We considered male-male competition as the main driver of sexual selection and group spawning in this prolonged mating species because the cost of choosing mates with specific genetic traits may be high for females. Male-male competition and female mate choice are the main drivers of animal sexual selection and mating behaviors. However, their impacts on the evolution of mating system are not yet quite clear. By studying the Omei treefrog, a Chinese endemic anuran species with common group spawning behaviors, this research posed the importance of male-male competition and female mate choice and discussed the mechanisms of sexual selection and multiple mating, which are the most essential and debated issues in evolutionary biology and behavioral ecology. We found that, in this prolonged mating and lek-patterned species, male-male competition is the main driver of sexual selection and group spawning. Larger males can get larger females, have more breeding opportunities, occupy better amplectant positions, and thus, obtain greater numbers of offspring. Whereas, females have no significant preferences on ornamentation and genetic traits of their males or mating positions, and the inferior males “make the best of a bad lot” by joining mating pairs to produce mating groups. Our study provides empirical evidence of reproductive mechanisms of amphibian species and could advance the understandings on the evolution of animals’ sexual selection and mating system.

Female polyandry and corresponding multiple paternity have been detected in many species. Several hypotheses, such as the fertility guarantee hypothesis, the good gene hypothesis, the genetic compatibility hypothesis and the genetic diversity hypothesis, have been proposed to explain the evolution of female polyandry. Anuran amphibians are widely observed to breed in polyandrous spawning groups; however, few studies have evaluated multiple paternity in these species or the evolutionary mechanisms underlying polyandry. In this study, we addressed these questions pertaining to polyandry in the Omei treefrog, Rhacophorus omeimontis. Eight microsatellite markers of R. omeimontis were used to determine the patterns of paternity. Paternity analyses revealed that offspring were multiply sired in a majority (79.07%) of the polyandrous spawning groups. A male's reproductive output during a breeding season was greatly affected by the mating rate, revealing that male fitness benefited from multiple copulations. By contrast, no evidence supported the fertility guarantee hypothesis that polyandry increases the fertility rate of females or the compatible/good gene hypotheses that polyandrous females produce more compatible or superior offspring. The increased allelic diversity observed in polyandrous clutches is probably a nonadaptive by-product of multiple mating. In conclusion, males join as many copulations as possible to increase their reproductive success, regardless of whether their mate has already been in amplexus with other males. Females may gain no benefits from polyandry but are incapable of preventing superfluous males from joining the mating group due to the physical characteristics and breeding behaviour of anurans. Thus, in R. omeimontis, male pursuit of higher reproductive success is the primary explanation for female polyandry.

Species richness, endemism, and threat status represent different biodiversity attributes important in identifying biodiversity hotspots and conservation priorities. Global distributions of areas with the most species overall, most endemic species, and most threatened species are often not coincident. Previous studies have only considered single patterns or have considered patterns separately, such that an integrated biodiversity index is needed for biodiversity assessment and conservation planning. Based on a comprehensive database of amphibian distributions in China, we analyzed the congruence of species richness (SR), endemic species richness (EMSR), IUCN threatened species richness (THSR), proportion of endemic species (%EM), and proportion of IUCN threatened species (%TH). To identify conservation priorities and protection gaps for Chinese amphibians, we explored an integrated indicator that simultaneously considered the five biodiversity patterns using principal component analyses. Although we found significant nonlinearities among SR, EMSR, and THSR patterns, their relationships to %EM and %TH were weak, and overlaps in hotspots of the five biodiversity types were limited. Our results showed that amphibian biodiversity hotspots are focused in southern, southwestern, and central China, and that the national protected area network is not effective in representing amphibian species distributions and addressing amphibian conservation priorities in China. The network is particularly ineffective in the south, which has the highest biodiversity in China, but small and insufficient protected areas. Policy, governance, and the protected area system must be revised and improved. We provide an integrated biodiversity indicator that can provide a reference for conservation priority setting for different taxa in other areas.

To explore the effect of hypoxia on the cell proliferation and expression of hypoxia-inducible factor-1α(HIF-1α) of human leukemia K562 cells.The leukemia cells were divided into 2 groups: hypoxia-treated group and conventional oxygen group as control. The K562 cells in hypoxia-treated group were treated with 50, 200, 400 and 800 µmol/L of CoClThe cell morphology was changed with increase of CoClThe hypoxia can inhibit the proliferation of K562 cells and up-requlate the expression of HIF-1α in cells, but does not induce cell differentiation.

Across taxa, females are routinely choosier than males in selecting mates. Several hypotheses have been advanced to explain genetic benefits behind female strategies. The inbreeding avoidance hypothesis suggests that females avoid mating with close relatives, thereby avoiding the matchup of deleterious recessive alleles. Outbreeding avoidance hypothesis suggests that females should not mate with too distantly related individuals so as to avoid the breakup of coadapted gene complexes. Although previous studies have suggested that selection should favor individuals that optimize the balance between inbreeding and outbreeding, detailed research is necessary to document the trade-off between them and variability in mate choice across a gradient of inbreeding levels among populations. The good genes and the genetic compatibility hypotheses predict that females choose mates according to costly traits and genetic dissimilarity, respectively. Thus, to document inbreeding or outbreeding depressions and assess the contributions of mate choice based upon good genes versus genetic compatibility, we examined egg production, collected body length measurements and genotyped five microsatellite markers in six populations of Asiatic toad (Bufo gargarizans). Our results revealed that the incidence of inbreeding was higher than that expected under the assumption of random mating and relatedness between mated individuals increased when the average inbreeding level increased among populations. Our findings did not support the good genes or the genetic compatibility hypotheses. Although some other processes could have influences on mate choice of Asiatic toad and need to be tested, our results indicated that, in small and isolated toad populations, the limited availability and high cost of obtaining unrelated mates may promote outbreeding avoidance and adaptation to inbreeding to be the critical drives of female mate choice.

Pashinintide A (1), a new cyclic hexapeptide, included a saccharose, and a new cyclic heptapeptide named pashinintide B (2) were isolated from the branches and leaves of Pyrus pashia Buch.-Ham. ex D. Don. 1 and 2 present the first two plant cyclopeptides from Rosaceae. And the stable complex of cyclic hexapeptide and sucrose in 1 suggests that cyclic hexapeptide might be a new natural receptor for saccharide.

Understanding the effects of climate change on primate ranging patterns is crucial for conservation planning. Rhinopithecus roxellana is an endangered primate species distributed in mountainous forests at the elevation of 1500–3500 m a.s.l. in China. Our study site, the Shennongjia National Nature Reserve, represents the eastern-most distribution of this species. This area has experienced significant habitat loss and fragmentation because of human population growth, increased farming and logging, and climate change. To estimate how changes in temperature and rainfall will affect the presumed future distribution of this species, we examined eco-geographic factors including bioclimate, habitat (vegetation type, landcover, etc.), topography, and human impact (human population, gross domestic product, etc.), and provide suggestions for management and conservation. We used a maximum entropy approach to predict the location and distribution of habitats suitable for R. roxellana in the present, 2020, 2050, and 2080 based on 33 environmental parameters, three general circulation models, three emissions scenarios, and two dispersal hypotheses. According to the ensemble modeling, we found range reductions of almost 30% by 2020, 70% by 2050, and over 80% by 2080. Although no obvious differences were found in distribution change based on full and zero dispersal assumptions, our results revealed range reductions in response to elevational, latitudinal, and longitudinal gradients, with the monkeys forced to migrate to higher elevations over time. Bioclimte factors, such as temperature, precipitation, evapo-transpiration, and aridity condition, were dominant contributors to range shifting. As habitat loss due to human influence and climate change is likely to be even more severe in the future, we considered three conservation hot-spots in the Shennongjia area and recommended: (i) securing existing reserves and establishing new reserves, (ii) re-designing management systems to include the Shenongjia reserve and the surrounding reserves and highlighting ecosystem protection at higher elevations, and (iii) using finer-scale research to guide the conservation planning and education in order to enhance protection and awareness in the local community. National and provincial conservation policies should integrate projections of climate change in making effective conservation strategies. Am. J. Primatol. 77:135–151, 2015. © 2014 Wiley Periodicals, Inc.

In this paper, we discuss the upper and lower bounds of unknown functions in several nonlinear integral inequalities. Firstly, we give out the upper estimation of unknown function of a nonlinear integral inequality. Secondly, we give out the lower estimation of unknown function of another nonlinear integral inequality. Finally, we discuss the upper and lower bounds of a linear integral inequality by adopting novel analysis techniques, such as change of variable, amplification method, differential and integration.

Based on the elementary method, the formulas of the probability distribution of the number of runs and its mathematical expectation and variance are proposed. Furthermore, an application of the number of runs in biological medicine testing is discussed.

In this paper, we discuss a class of linear integral equation with piecewise continuous function. Firstly, we change the integral equation to a differential equation with the initial condition. Secondly, the differential equation is solved by the constant variation formula and integration by parts. Explicit solution of the integral equation is given clearly.

Sexual selection is critical to both reproduction and evolution. The effects of male–male competition and female choice regarding body size have been examined in a large number of taxa, including toad species. Males and females have different optimal reproductive strategies, achieving breeding advantages in discrepant ways. Further, the relative contributions of intra- and inter-sexual size selection vary among species. Thus, to understand the mechanisms affecting mating success, it is important to consider both male–male and male–female interactions simultaneously and elucidate their interrelationship. In this study, we measured body sizes of all mated and unmated individuals in a population of Asiatic toad (Bufo gargarizans) and counted fertilized eggs of several clutches. Based on correlation and regression techniques, we tested for female choice and intra-sexual competition among males relative to body size gradients, and we compared the relative importance of these two processes for mating success. Our results reveal that male–male competition and female choice simultaneously contribute to sexual selection in toads. Furthermore, both interactions are most intense among smaller toads. The synergistic trends of male–male competition and female choice support the mutual mate choice hypothesis and works to stabilize body size in B. gargarizans. Normal distributions of breeding success relative to body size were detected for both sexes, suggesting that medium-sized individuals enjoy a reproductive advantage in the population studied.

Animal skin, which is the tissue that directly contacts the external surroundings, has evolved diverse functions to adapt to various environments. Amphibians represent the transitional taxon from aquatic to terrestrial life. Exploring the molecular basis of their skin function and adaptation is important to understand the survival and evolutionary mechanisms of vertebrates. However, comprehensive studies on the molecular mechanisms of skin functions in amphibians are scarce. In this study, we sequenced the skin transcriptomes of seven anurans belonging to three families and compared the similarities and differences in expressed genes and proteins. Unigenes and pathways related to basic biological processes and special functions, such as defense, immunity and respiration, were enriched in functional annotations. A total of 108 antimicrobial peptides were identified. The highly expressed genes were similar in species of the same family but were different among families. Additionally, the positively selected orthologous groups were involved in biosynthesis, metabolism, immunity and defense processes. This study is the first to generate extensive transcriptome data for the skin of seven anurans and provides unigenes and pathway candidates for further studies on amphibian skin function and adaptation.

A nitro aromatic explosives sensor based on fluorescence quenching was studied in this paper. U-shaped plastic optical fiber was used as sensor head and light emitting conjugated polymer MEH-PPV as fluorescence indictor. The light source was modulated sinusoidal to eliminate the external light disturbance. TNT quenching characteristics of fluorescence intensity were measured; When TNT concentration was lower than 10mg/ml, the relationship between fluorescence intensity and TNT concentration was linearly, when TNT concentration was higher than 10mg/100ml the obtained characteristics showed deviation from linear relation described by Stern-Volmer equation.

In order to achieve a thorough coverage of the basal lineages in the Chinese matrilineal pool, we have sequenced the mitochondrial DNA (mtDNA) control region and partial coding region segments of 6,093 mtDNAs sampled from 84 populations across China. By comparing with the available complete mtDNA sequences, 194 of those mtDNAs could not be firmly assigned into the available haplogroups. Completely sequencing 51 representatives selected from these unclassified mtDNAs identified a number of novel lineages, including five novel basal haplogroups that directly emanate from the Eurasian founder nodes (M and N). No matrilineal contribution from the archaic hominid was observed. Subsequent analyses suggested that these newly identified basal lineages likely represent the genetic relics of modern humans initially peopling East Asia instead of being the results of gene flow from the neighboring regions. The observation that most of the newly recognized mtDNA lineages have already differentiated and show the highest genetic diversity in southern China provided additional evidence in support of the Southern Route peopling hypothesis of East Asians. Specifically, the enrichment of most of the basal lineages in southern China and their rather ancient ages in Late Pleistocene further suggested that this region was likely the genetic reservoir of modern humans after they entered East Asia.

In this work, the electrochemical behavior of ferrocene (Fc) was investigated by cyclic voltammetry (CV) in room temperature ionic liquids (RTILs) of l-ethyl-3-methylimidazolium tetrafluoroborate (EMIBF 4 ) on glass carbon (GC), edge plane pyrolytic graphite (EPPG) and multi-walled carbon nanotube (MWCNTs)-modified EPPG electrodes, respectively. The results demonstrated that on GC electrode, pairs of well-defined reversible peaks were observed, while for the electrode of EPPG, the peak potential separation (AEp) is obviously larger than the theoretical value of 59 mV, hinting that the electrode of EPPG is distinguished from the commonly used electrode, consistent with the previous proposition that EPPG has many "defects". To obtain an improved electrochemical response, multi-walled carbon nanotubes (MWCNTs) were modified on the electrode of EPPG; the increased peak current and promoted peak potential separation not only proved the existence of "defects" in MWCNTs, but also supported that "creating active points" on an electrode is the main contribution of MWCNTs. Initiating the electrochemical research of Fc on the MWCNTs-modified EPPG electrode in RTILs and verifying the presence of "defects" on both EPPG and MWCNTs using cyclic voltammograms (CVs) of Fc obtained in RTILs of EMIBF 4 , is the main contribution of this preliminary work.

Threatened species typically have a small or declining population size, which make them highly susceptible to loss of genetic diversity through genetic drift and inbreeding. Genetic diversity determines the evolutionary potential of a species; therefore, maintaining the genetic diversity of threatened species is essential for their conservation. In this study, we assessed the genetic diversity of the adaptive major histocompatibility complex (MHC) genes in an endangered and narrowly distributed amphibian species, Leptobrachium leishanense in Southwest China. We compared the genetic variation of MHC class I genes with that observed in neutral markers (5 microsatellite loci and cytochrome b gene) to elucidate the relative roles of genetic drift and natural selection in shaping the current MHC polymorphism in this species. We found a high level of genetic diversity in this population at both MHC and neutral markers compared with other threatened amphibian species. Historical positive selection was evident in the MHC class I genes. The higher allelic richness in MHC markers compared with that of microsatellite loci suggests that selection rather than genetic drift plays a prominent role in shaping the MHC variation pattern, as drift can affect all the genome in a similar way but selection directly targets MHC genes. Although demographic analysis revealed no recent bottleneck events in L. leishanense, additional population decline will accelerate the dangerous status for this species. We suggest that the conservation management of L. leishanense should concentrate on maximizing the retention of genetic diversity through preventing their continuous population decline. Protecting their living habitats and forbidding illegal hunting are the most important measures for conservation of L. leishanense.

Understanding the effects of climate change on primate ranging patterns is crucial for conservation planning. Rhinopithecus roxellana is an endangered primate species distributed in mountainous forests at the elevation of 1500-3500 m a.s.l. in China. Our study site, the Shennongjia National Nature Reserve, represents the eastern-most distribution of this species. This area has experienced significant habitat loss and fragmentation because of human population growth, increased farming and logging, and climate change. To estimate how changes in temperature and rainfall will affect the presumed future distribution of this species, we examined eco-geographic factors including bioclimate, habitat (vegetation type, landcover, etc.), topography, and human impact (human population, gross domestic product, etc.), and provide suggestions for management and conservation. We used a maximum entropy approach to predict the location and distribution of habitats suitable for R. roxellana in the present, 2020, 2050, and 2080 based on 33 environmental parameters, three general circulation models, three emissions scenarios, and two dispersal hypotheses. According to the ensemble modeling, we found range reductions of almost 30% by 2020, 70% by 2050, and over 80% by 2080. Although no obvious differences were found in distribution change based on full and zero dispersal assumptions, our results revealed range reductions in response to elevational, latitudinal, and longitudinal gradients, with the monkeys forced to migrate to higher elevations over time. Bioclimte factors, such as temperature, precipitation, evapo-transpiration, and aridity condition, were dominant contributors to range shifting. As habitat loss due to human influence and climate change is likely to be even more severe in the future, we considered three conservation hot-spots in the Shennongjia area and recommended: (i) securing existing reserves and establishing new reserves, (ii) re-designing management systems to include the Shenongjia reserve and the surrounding reserves and highlighting ecosystem protection at higher elevations, and (iii) using finer-scale research to guide the conservation planning and education in order to enhance protection and awareness in the local community. National and provincial conservation policies should integrate projections of climate change in making effective conservation strategies.

Background Comparison of major histocompatibility complex (MHC) genes across vertebrate species can reveal molecular mechanisms underlying the evolution of adaptive immunity-related proteins. As the first terrestrial tetrapods, amphibians deserve special attention because of their exposure to probably increased spectrum of microorganisms compared with ancestral aquatic fishes. Knowledge regarding the evolutionary patterns and mechanisms associated with amphibian MHC genes remains limited. The goal of the present study was to isolate MHC class I genes from two Rhacophoridae species (Rhacophorus omeimontis and Polypedates megacephalus) and examine their evolution. Results We identified 27 MHC class I alleles spanning the region from exon 2 to 4 in 38 tree frogs. The available evidence suggests that these 27 sequences all belong to classical MHC class I (MHC Ia) genes. Although several anuran species only display one MHC class Ia locus, at least two or three loci were observed in P. megacephalus and R. omeimontis, indicating that the number of MHC class Ia loci varies among anuran species. Recombination events, which mainly involve the entire exons, played an important role in shaping the genetic diversity of the 27 MHC class Ia alleles. In addition, signals of positive selection were found in Rhacophoridae MHC class Ia genes. Amino acid sites strongly suggested by program to be under positive selection basically accorded with the putative antigen binding sites deduced from crystal structure of human HLA. Phylogenetic relationships among MHC class I alleles revealed the presence of trans-species polymorphisms. Conclusions In the two Rhacophoridae species (1) there are two or three MHC class Ia loci; (2) recombination mainly occurs between the entire exons of MHC class Ia genes; (3) balancing selection, gene duplication and recombination all contribute to the diversity of MHC class Ia genes. These findings broaden our knowledge on the evolution of amphibian MHC systems.

Objective To explore the relationship between nesfatin-1 and growth and development in newborns. Methods Blood samples for nesfatin-1, ghrelin, insulinlike growth factor-1 (IGF-1), insulin and glucose were obtained from preterm (n = 53) and term infants (n = 60), including appropriate for gestational age (AGA) (n = 32) and small for gestational age (SGA) infants (n = 28). The relationship between nesfatin-1 and other metabolic hormones or anthropometric parameters was evaluated. Results The concentrations of nesfatin-1, ghrelin and insulin and the homeostasis model assessment-insulin resistance index (HOMA-IR) were higher in SGA than AGA infants (p = 0.0358, 0.0163, 0.0001 and 0.0051, respectively), but IGF-1 levels and homeostasis model assessment-insulin sensitivity index (HOMA-ISI) were lower (p = 0.033 and 0.0001, respectively). Nesfatin-1 levels in SGA infants were higher on postnatal day 0 (PNDO) than in AGA infants (p = 0.0358) and lower on PND7 (p = 0.0002) and PND28 (p = 0.0488). A negative correlation showed between nesfatin-1 and oral calorie intake (r = -0.446; p = 0.017) and HOMA-ISI (r = -0.398; p = 0.036), and a positive correlation between nesfatin-1 and HOMA-IR (r = 0.43; p = 0.023) in SGA infants. Conclusion Nesfatin-1 is involved in the physiological regulation of intrauterine and postnatal growth and development in SGA infants.

Himalayas was believed to be a formidably geographical barrier between South and East Asia. The observed high frequency of the East Eurasian paternal lineages in Nepal led some researchers to suggest that these lineages were introduced into Nepal from Tibet directly; however, it is also possible that the East Eurasian genetic components might trace their origins to northeast India where abundant East Eurasian maternal lineages have been detected. To trace the origin of the Nepalese maternal genetic components, especially those of East Eurasian ancestry, and then to better understand the role of the Himalayas in peopling Nepal, we have studied the matenal genetic composition extensively, especially the East Eurasian lineages, in Nepalese and its surrounding populations. Our results revealed the closer affinity between the Nepalese and the Tibetans, specifically, the Nepalese lineages of the East Eurasian ancestry generally are phylogenetically closer with the ones from Tibet, albeit a few mitochondrial DNA haplotypes, likely resulted from recent gene flow, were shared between the Nepalese and northeast Indians. It seems that Tibet was most likely to be the homeland for most of the East Eurasian in the Nepalese. Taking into account the previous observation on Y chromosome, now it is convincing that bearer of the East Eurasian genetic components had entered Nepal across the Himalayas around 6 kilo years ago (kya), a scenario in good agreement with the previous results from linguistics and archeology.

We agree with van Oven (1) that a uniform mitochondrial (mt)DNA haplogroup nomenclature is indispensable for the future studies, and we actually propagated the same idea a few years ago (2). The current conflicts regarding the Asian haplogroup nomenclature (1) were essentially caused by publication delay: at the time when we submitted the manuscript (July 14, 2009), there was no conflict between the haplogroup system in our paper (3) and PhyloTree (mtDNA tree Build 5; http://www.phylotree.org/) to which we referred. To avoid similar problems in the future, a possible solution is that researchers in the mtDNA field indicate to van Oven (1) on submission of a paper that certain haplogroup names are being introduced, so that other researchers would at least not choose the same code for their newly identified haplogroups. The blocked names (possibly with some further information) could then be shown in the bulk mtDNA tree on that website.

Gelao ethnic group, an aboriginal population residing in southwest China, has undergone a long and complex evolutionary process. To investigate the genetic structure of this ancient ethnic group, mitochondrial DNA (mtDNA) polymorphisms of 102 Gelao individuals were collected and analyzed in this study. With the aid of the information extracted from control-region hypervariable segments (HVSs) I and II as well as some necessary coding-region segments, phylogenetic status of all mtDNAs under study were determined by means of classifying into various defined haplogroups. The southern-prevalent haplogroups B, R9, and M7 account for 45.1% of the gene pool, whereas northern-prevalent haplogroups A, D, G, N9, and M8 consist of 39.2%. Haplogroup distribution indicates that the Gelao bears signatures of southern populations and possesses some regional characters. In the PC map, Gelao clusters together with populations with Bai-Yue tribe origin as well as the local Han and the Miao. The results demonstrate the complexity of Gelao population and the data can well supplement the China mtDNA database.

Due to its numerous environmental extremes, the Tibetan Plateau—the world's highest plateau—is one of the most challenging areas of modern human settlement. Archaeological evidence dates the earliest settlement on the plateau to the Late Paleolithic, while previous genetic studies have traced the colonization event(s) to no earlier than the Neolithic. To explore whether the genetic continuity on the plateau has an exclusively Neolithic time depth, we studied mitochondrial DNA (mtDNA) genome variation within 6 regional Tibetan populations sampled from Tibet and neighboring areas. Our results confirm that the vast majority of Tibetan matrilineal components can trace their ancestry to Epipaleolithic and Neolithic immigrants from northern China during the mid-Holocene. Significantly, we also identified an infrequent novel haplogroup, M16, that branched off directly from the Eurasian M founder type. Its nearly exclusive distribution in Tibetan populations and ancient age (>21 kya) suggest that M16 may represent the genetic relics of the Late Paleolithic inhabitants on the plateau. This partial genetic continuity between the Paleolithic inhabitants and the contemporary Tibetan populations bridges the results and inferences from archaeology, history, and genetics.

We aimed to reduce the incidence of seroma formation by altering surgical technique.Two hundred one breast cancer patients were randomly divided into 2 arms: arm 1 was operated on using an altered surgical technique, which is to ligate all of the tissue connecting axillary vein bundles to the specimen, to suture the anterior edge of the latissimus dorsi to the chest wall, and to fix the skin flap to the underlying muscle by subcutaneous sutures; arm 2 was operated on using the conventional technique.The drainage volume, in the initial 3 days, for patients in arm 1 was significantly less than that for patients in arm 2 (P.01). The duration of drainage in arm 1 was shorter than that in arm 2 (P.01). The incidence of seroma formation in arm 1 (2%) was significantly less than that in arm 2 (14%) (P.01).The modified operating technique is an effective approach to reducing the incidence of seroma formation after mastectomy and axillary dissection.

The adaptive multi-rate (AMR) is the mandatory speech codec for GSM system, ranging from 12.2 kbps down to 4.75 kbps. To satisfy the requirement of need for both audio and video simultaneously, a kind of hardware and software of AMR speech encoder based on high-performance chips TMS320C64X DSP is proposed. Considering the limited resource of DSP chips, the optimization scheme is referred to later. The result shows that the CPU-load is reduced from 38 percent to 4 percent. And the time of processing each frame is also reduced greatly after optimizing. It is verified with all the test vectors provided by 3 GPP, and the stable operation on the real-time testing board was also confirmed. So, more resource is reserved for video coding on DSP chips. This is helpful for implementing both speech coder and video coder on one DSP chip.

We examined the penetrability of pig, rat and bull spermatozoa into zona-free pig oocytes. Frozen-thawed boar spermatozoa penetrated into both zona-intact and zona-free oocytes with similar efficacy in a modified Tris-buffered medium (mTBM) supplemented with BSA and caffeine, but not in medium without caffeine. Rat epididymal spermatozoa did not readily penetrate into zona-free pig oocytes in mTBM with BSA. However, when a modified Krebs-Ringer bicarbonate solution was used, penetration rate varied with sperm concentrations at insemination: 79% of the oocytes were penetrated at 1.0 x 10(6) cells/ml, but very few at 0.1 x 10(6) and 10.0 x 10(6) cells/ml. In all oocytes penetrated, no activation was observed and the sperm nucleus was fully decondensed but did not transform into a male pronucleus. Frozen-thawed bull spermatozoa were also found to penetrate into zona-free pig oocytes in mTBM with BSA, caffeine and heparin: higher penetration rates were obtained with 1.0 x 106 and 10.0 x 10(6) spermatozoa/ml compared with 0.1 x 10(6) spermatozoa/ml. The penetration rate with 1.0 x 10(6) spermatozoa/ml was stable in five different bulls. All oocytes penetrated were activated and male pronuclear formation was observed in 57-79% of the penetrated oocytes. These results suggest that capacitation or the acrosome reaction is required for boar, rat, and possibly, bull spermatozoa to penetrate into zona-free pig oocytes. Bull spermatozoa can easily induce activation of pig oocytes and form male pronuclei, but rat spermatozoa cannot do so, indicating species differences in the ability of spermatozoa to activate pig oocytes and to transform to male pronuclei in the ooplasm.

Background Archaeological studies have revealed a series of cultural changes around the Last Glacial Maximum in East Asia; whether these changes left any signatures in the gene pool of East Asians remains poorly indicated. To achieve deeper insights into the demographic history of modern humans in East Asia around the Last Glacial Maximum, we extensively analyzed mitochondrial DNA haplogroup M9a'b, a specific haplogroup that was suggested to have some potential for tracing the migration around the Last Glacial Maximum in East Eurasia. Results A total of 837 M9a'b mitochondrial DNAs (583 from the literature, while the remaining 254 were newly collected in this study) pinpointed from over 28,000 subjects residing across East Eurasia were studied here. Fifty-nine representative samples were further selected for total mitochondrial DNA sequencing so we could better understand the phylogeny within M9a'b. Based on the updated phylogeny, an extensive phylogeographic analysis was carried out to reveal the differentiation of haplogroup M9a'b and to reconstruct the dispersal histories. Conclusions Our results indicated that southern China and/or Southeast Asia likely served as the source of some post-Last Glacial Maximum dispersal(s). The detailed dissection of haplogroup M9a'b revealed the existence of an inland dispersal in mainland East Asia during the post-glacial period. It was this dispersal that expanded not only to western China but also to northeast India and the south Himalaya region. A similar phylogeographic distribution pattern was also observed for haplogroup F1c, thus substantiating our proposition. This inland post-glacial dispersal was in agreement with the spread of the Mesolithic culture originating in South China and northern Vietnam.