Comparative assessment of the intragenomic variations of dinoflagellate Tripos species through single-cell sequencing.

Tripos is a large dinoflagellate genus widely distributed in the world's oceans. Morphology-based species identification is inconclusive due to high morphological intraspecific variability. Metabarcoding analysis has been demonstrated to be effective for species identification and tracking their spatiotemporal dynamics. However, accumulating evidence suggests high levels of intragenomic variations (IGVs) are common in many algae, leading to concerns about overinterpretation of molecular diversity in metabarcoding studies. In this project, we evaluated and compared IGVs in Tripos species by conducting the first high-throughput sequencing (HTS) of 18S rDNA V4 of Tripos single cells. High numbers of haplotypes (19-172) were identified in each of the 30 Tripos cells. Each cell contained one dominant haplotype with high relative abundance and many haplotypes with lower abundances. Thus, the presence of multiple minor haplotypes substantially overestimate the molecular diversity identified in metabarcoding analysis, which encompass not only interspecific and intraspecific diversities, but high levels of IGVs.

A neural m6A pathway regulates behavioral aggregation in migratory locusts.

RNA N6-methyladenosine (m6A), as the most abundant modification of messenger RNA, can modulate insect behaviors, but its specific roles in aggregation behaviors remain unexplored. Here, we conducted a comprehensive molecular and physiological characterization of the individual components of the methyltransferase and demethylase in the migratory locust Locusta migratoria. Our results demonstrated that METTL3, METTL14 and ALKBH5 were dominantly expressed in the brain and exhibited remarkable responses to crowding or isolation. The individual knockdown of methyltransferases (i.e., METTL3 and METTL14) promoted locust movement and conspecific attraction, whereas ALKBH5 knockdown induced a behavioral shift toward the solitary phase. Furthermore, global transcriptome profiles revealed that m6A modification could regulate the orchestration of gene expression to fine tune the behavioral aggregation of locusts. In summary, our in vivo characterization of the m6A functions in migratory locusts clearly demonstrated the crucial roles of the m6A pathway in effectively modulating aggregation behaviors.

The high molecular diversity in Noctiluca scintillans is dominated by intra-genomic variations revealed by single cell high-throughput sequencing of 18S rDNA V4.

The application of high-throughput sequencing (HTS) technologies has revolutionized research on phytoplankton biodiversity by generating an unprecedented amount of molecular data in marine ecosystem surveys. However, high-level of molecular diversity uncovered in HTS-based metabarcoding analyses may lead to overinterpretation of phytoplankton diversity due to excessive intra-genomic variations (IGVs). The aims in this study are to explore the nature of phytoplankton molecular diversity and to test the hypothesis. We carried out single-cell metabarcoding analysis of 18S rDNA V4 sequences obtained in single Noctiluca scintillans cells isolated from various sites in coastal waters of China. Results showed that each single N. scintillans cell harbored a high level of IGVs with about 100 amplicon sequence variants (ASVs). The large numbers of non-dominant ASVs identified in N. scintillans cells, which might correspond to the larger numbers of ASVs annotated as N. scintillans and showed similar temporal dynamics in metabarcoding analyses, could inflate the inter-species diversity or intra-species genetic diversity. In addition, there were large numbers of additional ASVs that were not annotated as N. scintillans. These non-N. scintillans ASVs might represent diverse preys for N. scintillans, consistent with previous reports that N. scintillans may act as chance predator of a broad-spectrum preys. This single-cell study has unambiguously demonstrated that the existence of high levels of IGVs in N. scintillans and most likely many other phytoplankton species, demonstrating that the majority of the molecular diversity revealed in metabarcoding analysis, which were generally interpreted as the sum of inter-species diversity and intra-species diversity, actually included high levels of IGVs and should be interpreted with caution.

Significance of the antibody orientation for the lateral flow immunoassays: A mini-review.

Lateral flow immunoassays (LFIAs) are well-established and broadly commercialized tools in the field of point-of-care testing due to their simplicity, rapidity, cost-effectiveness, and low requirements for users and equipment. However, the insensitivity and the possibility of producing inaccurate results associated with conventional LFIAs have impeded their wide-ranging implementation, especially for monitoring ultra-trace level of analytes. Moreover, the heterogeneous distribution of amino acids on the surface of antibody (Ab) results in a lack of precise control over their orientation, which ultimately leads to unsatisfactory detection performance. To address those concerns, herein we provide an overview of the emerging efforts to prepare well-established LFIAs from the perspective of orientation manipulation of immobilized Abs on the nanoprobes or membranes. The preparation of excellent nanoprobes with Abs being oriented immobilized, consisting of the nanoprobe types, Ab types, and their conjugation chemistries, are reviewed. Followed by the introduction of efforts highlight the importance of directionally immobilized Ab on the membrane. The effects of Ab orientation on the analytical performance of LFIA platforms in terms of sensitivity, specificity, rapidity, reliability, cost-effectiveness, and stability are also summarized. Finally, the future development and challenges of Ab-oriented immobilization-assisted LFIAs are also discussed.

Impacts of oxygen deficiency on embryo life-history traits of migratory locust Locusta migratoria from low and high altitudes.

Hypoxia challenges aerobic organisms in numerous environments, and hypoxic conditions may become more severe under future climate-change scenarios. The impact of hypoxia on the development of terrestrial insect embryos is not well understood. Here, to address this gap, embryonic life-history traits of migratory locust Locusta migratoria from low-altitude and high-altitude regions were compared under 2 oxygen levels: normoxia (i.e., 21 kPa oxygen partial pressure and mild hypoxia (i.e., 10 kPa oxygen partial pressure). Our results demonstrated that, whether reared under normoxia or mild hypoxia, L. migratoria from high-altitude populations had longer developmental times, reduced weight, and lower mean relative growth rate as compared with those from low-altitude populations. When transferred from normoxia to mild hypoxia, nearly all the tested life-history traits presented significant negative changes in the low-altitude populations, but not in the high-altitude populations. The factor 'strain' alone explained 18.26%-54.59% of the total variation for traits, suggesting that the phenotypic differences between L. migratoria populations from the 2 altitudes could be driven by genetic variation. Significant genetic correlations were found between life-history traits, and most of these showed differentiation between the 2 altitudinal gradients. G-matrix comparisons showed significant structural differences between L. migratoria from the 2 regions, as well as several negative covariances (i.e., trade-offs) between traits in the low-altitude populations. Overall, our study provides clear evidence that evolutionary divergence of embryonic traits between L. migratoria populations from different altitudes has occurred.

The lead and cadmium content in rice and risk to human health in China: A systematic review and meta-analysis.

Numerous studies have investigated concentrations of lead (Pb) and cadmium (Cd) in rice in China, but have come to divergent conclusions. Therefore we systematically reviewed and meta-analyzed the available evidence on levels of Pb and Cd in rice in different regions of China in order to assess the potential risk to human health. The meta-analysis included 24 studies of Pb levels and 29 studies of Cd levels, published in 2011-2021. The pooled Pb concentration in rice was 0.10 mg per kg dry weight (95% CI 0.08-0.11), while the pooled Cd concentration was 0.16 mg per kg dry weight (95% CI 0.14-0.18). These levels are within the limits specified by national food safety standards. However, the total target hazard quotient for both metals exceeded 1.0 for adults and children, suggesting that rice consumption poses a health risk.

The FSHR G-29A variant is not associated with the ovarian response to exogenous FSH stimulation.

A common genetic variant in the follicle stimulating hormone receptor gene (FSHR) 5'-untranslated region has been previously reported to influence FSHR gene expression. However, studies on the ovarian response to exogenous gonadotropin stimulation are limited. The aim of this study was to evaluate the association of variants at positions -29 of the FSHR gene with the ovarian response to exogenous FSH stimulation in Chinese women. The genotypes of the FSHR gene were assayed using the Sequenom MassARRAY system. Total RNA and protein was extracted from granulosa cells, and FSHR expression at the mRNA and protein levels was assessed using quantitative PCR and western blotting. Our data revealed that there was no association between the FSHR genotype at the -29 position and the outcome of controlled ovarian stimulation. The expression of FSHR, at both the mRNA and protein levels, was similar amongst the different FSHR genotypes assessed, but was significantly reduced in the low responders. These results indicate that the variants caused by mutations at position -29 are not associated with ovarian response, and the low ovarian response to gonadotropin stimulation may be caused by decreased FSHR expression.

Association of Sperm Methylation at LINE-1, Four Candidate Genes, and Nicotine/Alcohol Exposure With the Risk of Infertility.

In this study, we examined whether smoking and drinking affect sperm quality and the DNA methylation of the repetitive element LINE-1, MEST, P16, H19, and GNAS in sperm. Semen samples were obtained from 143 male residents in a minority-inhabited district of Guizhou province in southwest China. Quantitative DNA methylation analysis of the samples was performed using MassARRAY EpiTYPER assays. Sperm motility was significantly lower in both the nicotine-exposed (P = 0.0064) and the nicotine- and alcohol-exposed (P = 0.0008) groups. Follicle-stimulating hormone (FSH) levels were higher in the nicotine-exposed group (P = 0.0026). The repetitive element LINE-1 was hypermethylated in the three exposed groups, while P16 was hypomethylated in the alcohol and both the alcohol and nicotine exposure groups. Our results also show that alcohol and nicotine exposure altered sperm cell quality, which may be related to the methylation levels of MEST and GNAS. In addition, MEST, GNAS, and the repetitive element LINE1 methylation was significantly associated with the concentration of sperm as well as FSH and luteinizing hormone levels.

Sequence variants in FSHR and CYP19A1 genes and the ovarian response to controlled ovarian stimulation.

OBJECTIVE: To examine whether sequence variants within the FSHR and CYP19A1 genes are related to the ovarian response to controlled ovarian stimulation (COS). DESIGN: Genetic association study using both single-gene and combined analyses of women with sequence variants undergoing in vitro fertilization treatment. SETTING: Academic research institute hospital. PATIENT(S): Seven hundred and five women undergoing ovarian stimulation with recombinant follicle-stimulating hormone (FSH). INTERVENTION(S): Peripheral blood extraction, DNA purification, and FSHR c.919G>A (rs6165, p.Thr307Ala) and CYP19A1 c.*19C>T (rs10046) sequence variants analyses. MAIN OUTCOME MEASURE(S): Single-gene statistical analysis and combined statistical analysis with the SPSS17.0 software; FSHR c.919G>A and CYP19A1 c.*19C>T sequence variant genotypes and clinical parameters related to the COS response as oocyte retrieval and hormone levels, doses of exogenous FSH. RESULT(S): Women with genotype Ala/Ala at FSHR position 307 had higher basal levels of FSH and were more likely to have a low ovarian response compared with other genotypes. Women with genotype TT at CYP19A1 yielded fewer oocytes after ovarian stimulation. The combined analysis of these two sequence variants revealed that these two single-nucleotide variants have a synergistic effect in conferring the risk of a low ovarian response. CONCLUSION(S): Our results support an association of sequence variants in the genes that participate in estrogen synthesis, notably the FSHR and CYP19A1 genes, with the outcome of COS.

Tideglusib, a prospective alternative to nonoxynol-9 contraceptive.

OBJECTIVES: We aimed to evaluate the antifertility activity and vaginal irritation effects of tideglusib in vivo using rabbit models and to evaluate the cytotoxical effects of tideglusib to sperm, vaginal cells and vaginal bacteria (L. acidophilus) in vitro. STUDY DESIGN: We treated female rabbits with vaginal tideglusib 1 mM, nonoxynol-9 (N-9) or vehicle control (Poloxamer 407). In experiment 1, we sacrificed females (n = 6 each) after 10 days of daily administration and assessed vaginal histological changes using Eckstein irritation score. In experiment 2, females (n = 9 each) received estradiol benzoate to induce ovulation 24 h prior to vaginal treatment followed by introduction of a fertile male. These females underwent necropsy at the 21st day to assess pregnancy status. In experiment 3, we used an HTM-TOX IVOS sperm motility analyzer and scanning electron microscopy (SEM) to evaluate the effect of tideglusib on human sperm samples. In experiment 4, we evaluated the effect of tideglusib on lactobacillus and vaginal cell growth in vitro. RESULTS: The total irritation score of tideglusib vs. N-9 was 3.4 ±â€¯2.07 vs. 7.8 ±â€¯3.82, p <.05. The pregnancy rate of tideglusib, N-9 and control group was 11.1%, 0% and 88.9%, respectively. Tideglusib exhibited a dose-dependent spermostatic/spermicidal activity, and the minimum effective concentrations of tideglusib and N-9 were 8.724 ±â€¯3.047 µM and 219.75 ±â€¯41.78 µM, respectively. SEM and transmission electron microscopy revealed acrosomal membrane impairments caused by tideglusib. Tideglusib was much less toxic to vaginal cells and L. acidophilus than N-9 in vitro. CONCLUSIONS: Evaluation using rabbit models indicated that tideglusib is a prospective spermicidal contraceptive with low vaginal irritation effects. IMPLICATIONS: Tideglusib or tideglusib analogues may be a contraceptive with perspective to replace N-9. It is possible for a spermicide to balance spermicidal activity and vaginal/cervical irritation effects very well.

Transcriptional Responses in Defense-Related Genes of Sitobion avenae (Hemiptera: Aphididae) Feeding on Wheat and Barley.

Many aphids can adapt on plants of differential resistance levels, but molecular interactions underlying host plant utilization and shift of aphids are still not well understood. Here, we sequenced the transcriptome and compared global gene expression profiles of Sitobion avenae (Fabricius) feeding on wheat (i.e., the susceptible plant) and barley (i.e., the resistant plant). In total, 65,200 high-quality unigenes were identified from the merged transcriptomes, and 861 of them were differentially expressed. Relative to their expression on barley, all differentially expressed P450 (cytochrome 450 monooxygenase) genes (e.g., SavCYP6A13, SavCYP4C1, and SavCYP4G15) in S. avenae on wheat were upregulated, suggesting the significance of P450s in xenobiotic metabolism for this aphid on wheat. For S. avenae on barley, some genes encoding for ATP-binding cassette (ABC) transporters (e.g., ABCG1, ABCG4, ABCB7, and ABCA5) and UDP-glucuronosyltransferases (UGTs; e.g., UGT2B1 and UGT2C1) showed a dramatic increase in expression, suggesting that ABC transporters and UGTs could be critical for detoxification metabolism in S. avenae on barley. In addition, the expression for nearly all differentially expressed proteases was upregulated for S. avenae on barley, reflecting xenobiotic challenges facing S. avenae on resistant barley. Thus, various detoxification and other defense-related genes (e.g., proteases and oxidases) showed highly inducible transcript regulation, suggesting great adaptive potential for S. avenae on different plants. All the abovementioned genes will be prime candidates for further studies of molecular interactions underlying host plant use and specialization in this aphid. Our results provide insights into aphids' defenses against variable phytochemicals, and the molecular basis underlying aphid-plant interactions as well.

Probing behaviors and their plasticity for the aphid Sitobion avenae on three alternative host plants.

Insects may develop different behavioral phenotypes in response to heterogeneous environments (e.g., host plants), but the plasticity of their feeding behaviors has been rarely explored. In order to address the issue, clones of the English grain aphid, Sitobion avenae (Fabricius), were collected from wheat, and their probing behaviors were recorded on three plants. Our results demonstrated that S. avenae individuals on the alternative plants (i.e., barley and oat) tended to have higher frequency of non-probing (Np), increased duration of the pathway phase, increased phloem salivation, and decreased phloem ingestion (E2), compared to those on the source plant (i.e., wheat), showing the resistance of barley and oat to this aphid's feeding. This aphid showed apparently high extents of plasticity for all test probing behaviors on barley or oat. Positive selection for higher extents of plasticity in E2 duration was identified on barley and oat. The factor 'clone' alone explained 30.6% to 70.1% of the total variance for each behavioral plasticity, suggesting that the divergence of probing behavior plasticity in S. avenae had a genetic basis. This aphid's fitness correlated positively with the plasticity of Np frequency and E2 frequency. Some behaviors and their corresponding plasticities (e.g., the frequency of xylem ingestion and its plasticity) were found to be correlated characters, probably reflecting the limits for the evolution of higher extents of behavioral plasticity in this aphid. The differential probing behaviors and their plasticity in S. avenae can have significant implications for the adaptation and management of aphids on different plants.

Effects of a presumably protective endosymbiont on life-history characters and their plasticity for its host aphid on three plants.

Hamiltonella defensa is well known for its protective roles against parasitoids for its aphid hosts, but its functional roles in insect-plant interactions are less understood. Thus, the impact of H. defensa infections on life-history characters and the underlying genetic variation for the grain aphid, Sitobion avenae (Fabricius), was explored on three plants (i.e., wheat, oat, and rye). Compared to cured lines, H. defensa infected lines of S. avenae had lower fecundity on wheat and oat, but not on rye, suggesting an infection cost for the aphid on susceptible host plants. However, when tested on rye, the infected lines showed a shorter developmental time for the nymphal stage than corresponding cured lines, showing some benefit for S. avenae carrying the endosymbiont on resistant host plants. The infection of H. defensa altered genetic variation underlying its host S. avenea's life-history characters, which was shown by differences in heritabilities and genetic correlations of life-history characters between S. avenae lines infected and cured of the endosymbiont. This was further substantiated by disparity in G-matrices of their life-history characters for the two types of aphid lines. The G-matrices for life-history characters of aphid lines infected with and cured of H. defensa were significantly different from each other on rye, but not on oat, suggesting strong plant-dependent effects. The developmental durations of infected S. avenae lines showed a lower plasticity compared with those of corresponding cured lines, and this could mean higher adaptability for the infected lines.Overall, our results showed novel functional roles of a common secondary endosymbiont (i.e., H. defensa) in plant-insect interactions, and its infections could have significant consequences for the evolutionary ecology of its host insect populations in nature.

A Self-Healing Aqueous Lithium-Ion Battery.

Flexible lithium-ion batteries are critical for the next-generation electronics. However, during the practical application, they may break under deformations such as twisting and cutting, causing their failure to work or even serious safety problems. A new family of all-solid-state and flexible aqueous lithium ion batteries that can self-heal after breaking has been created by designing aligned carbon nanotube sheets loaded with LiMn2 O4 and LiTi2 (PO4 )3 nanoparticles on a self-healing polymer substrate as electrodes, and a new kind of lithium sulfate/sodium carboxymethylcellulose serves as both gel electrolyte and separator. The specific capacity, rate capability, and cycling performance can be well maintained after repeated cutting and self-healing. These self-healing batteries are demonstrated to be promising for wearable devices.

Molecular and quantitative genetic differentiation in Sitobion avenae populations from both sides of the Qinling Mountains.

Quantitative trait differences are often assumed to be correlated with molecular variation, but the relationship is not certain, and empirical evidence is still scarce. To address this issue, we sampled six populations of the cereal aphid Sitobion avenae from areas north and south of the Qinling Mountains, and characterized their molecular variation at seven microsatellite loci and quantitative variation at nine life-history traits. Our results demonstrated that southern populations had slightly longer developmental times of nymphs but much higher lifetime fecundity, compared to northern populations. Of the nine tested quantitative characters, eight differed significantly among populations within regions, as well as between northern and southern regions. Genetic differentiation in neutral markers was likely to have been caused by founder events and drift. Increased subdivision for quantitative characters was found in northern populations, but reduced in southern populations. This phenomenon was not found for molecular characters, suggesting the decoupling between molecular and quantitative variation. The pattern of relationships between FST and QST indicated divergent selection and suggested that local adaptation play a role in the differentiation of life-history traits in tested S. avenae populations, particularly in those traits closely related to reproduction. The main role of natural selection over genetic drift was also supported by strong structural differences in G-matrices among S. avenae populations. However, cluster analyses did not result in two groups corresponding to northern and southern regions. Genetic differentiation between northern and southern populations in neutral markers was low, indicating considerable gene flow between them. The relationship between molecular and quantitative variation, as well as its implications for differentiation and evolution of S. avenae populations, was discussed.

The Ser680Asn polymorphism in the follicle-stimulating hormone receptor gene is associated with the ovarian response in controlled ovarian hyperstimulation.

OBJECTIVE: Polymorphisms in the follicle-stimulating hormone receptor (FSHR) gene are reported to be associated with the ovarian response in controlled ovarian hyperstimulation (COH), although there remains some discordance between studies. Here, using the largest patient sample to date, we evaluated the association of the p.Ser680Asn (S(680)N) polymorphism in the FSHR gene with the outcome of COH. DESIGN: Cohort study. SETTING: Medical academy and hospital. PATIENTS: A total of 1250 infertile Chinese women undergoing IVF/ICIS-ET treatment were included. MEASURES: The association between an FSHR polymorphism (S(680)N) and the ovarian response was analysed. Genotyping was performed by utilizing direct sequencing and the Sequenom MassARRAY iPLEX platform. Follicular fluid oestradiol (E2) and follicle-stimulating hormone (FSH) concentrations were determined using electrochemiluminesence immunoassays. The ovarian response parameters were analysed based on the FSHR genotypes. The odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated for the risk genotypes and alleles. RESULTS: There were linear correlations between the basal FSH level, exogenous gonadotropin consumption, and oocytes retrieved and the Ser680 alleles. Patients in the homozygous SS group demonstrated higher basal FSH levels, required more dosage of exogenous gonadotropin for ovarian stimulation, and had fewer numbers of oocytes retrieved compared with patients in the homozygous NN and heterozygous groups. Logistic regression analysis revealed that the OR of a poor ovarian response for the NS genotype was 1·79 (95% CI 1·28-2·61; P < 0·001), whereas that for the SS genotype was 2·25 (95% CI 1·40-3·58; P < 0·001) after adjusting for age, BMI and basal FSH level. The concentration of E2 in the follicular fluid was significantly higher in subjects with the NN genotype than the SS genotype (772 ± 545 ng/ml vs. 1299 ± 504 ng/ml). CONCLUSIONS: Follicle-stimulating hormone receptor gene polymorphism at position 680 is associated with different ovarian responses to controlled ovarian hyperstimulation.

Association of FTO polymorphisms with obesity and metabolic parameters in Han Chinese adolescents.

BACKGROUND: Previous studies have suggested that fat mass-and obesity-associated (FTO) gene is associated with body mass index (BMI) and the risk of obesity. This study aims to assess the association of five FTO polymorphisms (rs9939609, rs8050136, rs1558902, rs3751812 and rs6499640) with obesity and relative parameters in Han Chinese adolescents. METHODS: We examined a total of 401 adolescents, 223 normal weights (58.7% boys, 41.3% girls), 178 overweight (60.1% boys, 39.9% girls), aging from 14 to 18-years-old, recruited randomly from public schools in the central region of Wuxi, a southern city of China. DNA samples were genotyped for the five polymorphisms by Sequenom Plex MassARRAY. Association of the FTO polymorphisms with BMI, serum fasting plasm glucose (FPG), fasting insulin (FIns), triglyceride (TG) and cholesterol (TC) were investigated. RESULTS: 1) Serum FPG, FIns, TG and TC were statistically significant higher than that in normal control group. 2) We found that BMI was higher in the rs9939609 TA+AA, rs8050136 AC+AA, rs1558902 TA+AA and rs3751812 GT+TT genotypes than in wild TT genotypes (rs9939609: P = 0.038; rs1558902: P = 0.038;), CC genotypes(rs8050136: P = 0.024) and GG genotypes (rs3751812: P = 0.024), which were not significant on adjusting for multiple testing. 3) In case-control studies, five polymorphisms were not significantly associated with overweight (p>0.05), haplotype analyses showed non-haplotype is significantly associated with a higher risk of being overweight (p>0.05). 4) There existed no significant statistical difference about FPG, FIns, TG and TC in genotype model for any SNP. CONCLUSIONS: Our study has conducted a genetic association study of the FTO polymorphisms with BMI, serum fasting plasm glucose (FPG), fasting insulin (FIns), triglyceride (TG) and cholesterol (TC). Our study found BMI of subjects with A allele of FTO rs9939609 is higher than that with T allele. Further studies on other polymorphisms from FTO and increasing the sample size are needed.

Differential performance of Sitobion avenae populations from both sides of the Qinling Mountains under common garden conditions.

Sitobion avenae (F.) is a cosmopolitan cereal pest, but geographic barriers like the Qinling Mountains in the Shaanxi Province of China may lead to isolation among its populations, thus causing allopatric speciation. We sampled S. avenae populations from areas north (mean annual temperature, ≍9°C) and south (mean annual temperature, ≍14°C) of the Qinling Mountains, and tested them at 20°C in common garden experiments. The results showed that northern populations had reduced developmental time for first-instar nymphs but prolonged for third- and fourth-instar nymphs compared with southern populations. The postreproductive time and total lifespan of adults from southern populations were longer than those from northern populations, but no significant differences were found in reproductive time or age at first reproduction. Southern populations showed higher lifetime and daily fecundities than northern ones. Significant differences were found in correlation of life-history traits between northern and southern populations. Principal component analyses (PCAs) of S. avenae's vital life-history traits showed separation of populations from three southern locations, indicating their local adaptation. The clustering patterns generated by PCA also showed divergence between northern and southern populations. Alatae of S. avenae seemed to be able to disperse across the Qinling Mountains, which was indicated by the clustering together of some individuals from one side of the mountains with those from the other side. The impacts of the Qinling Mountains on the divergence and gene flow among S. avenae populations, as well as the potential of allopatric speciation for this species, are discussed.

Interleukin-8 regulates endothelial permeability by down-regulation of tight junction but not dependent on integrins induced focal adhesions.

Interleukin-8 (IL-8) is a common inflammatory factor, which involves in various non-specific pathological processes of inflammation. It has been found that increased endothelial permeability accompanied with high expression of IL-8 at site of injured endothelium and atherosclerotic plaque at early stages, suggesting that IL-8 participated in regulating endothelial permeability in the developing processes of vascular disease. The purpose of this study is to investigate the regulation effects of IL-8 on the vascular endothelial permeability, and the mRNA and protein expression of tight junction components (i.e., ZO-1, Claudin-5 and Occludin). Endothelial cells were stimulated by IL-8 with the dose of 50, 100 and 200 ng/mL, and duration of 2, 4, 6, 8h, respectively. The mRNA and protein expression level of tight junction components with IL-8 under different concentration and duration was examined by RT-PCR and Western blot, respectively. Meanwhile, the integrins induced focal adhesions event with IL-8 stimulation was also investigated. The results showed that IL-8 regulated the permeability of endothelium by down-regulation of tight junction in a dose- and time-dependence manner, but was not by integrins induced focal adhesions. This finding reveals the molecular mechanism in the increase of endothelial cell permeability induced by IL-8, which is expected to provide a new idea as a therapeutic target in vascular diseases.

The association of idiopathic recurrent pregnancy loss with polymorphisms in hemostasis-related genes.

Recurrent pregnancy loss (RPL) is a complex, multifactorial condition. Inherited thrombophilia is the leading cause of thromboembolism and is associated with an increased risk of RPL. The aims of the current study were to investigate the effects of polymorphisms in hemostasis-related genes antithrombin (SERPINC1), thrombomodulin (THBD), tissue factor pathway inhibitor (TFPI), factor V, factor II and annexin A5 (ANXA5), involved in reproductive failure in 94 RPL cases with two or more consecutive pregnancy losses prior to 20weeks of pregnancy and 169 healthy controls who had at least one term delivery and no history of pregnancy loss. The genotypes of SERPINC1 G786A, THBD C1418T, TFPI T-33C, factor V G1628A, factor II A19911G and ANXA5 G76A were assayed by the Sequenom MassARRAY system. Genotype and allele frequencies for SERPINC1 (rs2227589), TFPI (rs8176592), factor V (rs6020), factor II (rs3136516) and ANXA5 (rs113588187) in cases and controls were similar. The distribution of THBD C1418T allele showed significant differences between RPL cases and healthy controls (odds ratio (OR): 1.58, 95%, confidence interval (CI): 1.05-2.39, P=0.027). In univariate logistic regression analyses, carriers of THBD 1418T allele (CT+TT) had an increased risk of RPL (OR: 1.83, 95%, CI: 1.10-3.06, P=0.020). This indicated that THBD 1418T allele was associated with increasing the risk of RPL.