Serological survey of avian influenza virus infection in non-avian wildlife in Xinjiang, China.

We conducted a serological survey to detect antibodies against avian influenza virus (AIV) in Gazella subgutturosa, Canis lupus, Capreolus pygargus, Sus scrofa, Cervus elaphus, Capra ibex, Ovis ammon, Bos grunniens and Pseudois nayaur in Xinjiang, China. Two hundred forty-six sera collected from 2009 to 2013 were assayed for antibodies against H5, H7 and H9 AIVs using hemagglutination inhibition (HI) tests and a pan-influenza competitive ELISA. Across all tested wildlife species, 4.47 % harbored anti-AIV antibodies that were detected by the HI assay. The seroprevalence for each AIV subtype across all species evaluated was 0 % for H5 AIV, 0.81 % for H7 AIV, and 3.66 % for H9 AIV. H7-reactive antibodies were found in Canis lupus (9.09 %) and Ovis ammon (4.55 %). H9-reactive antibodies were found in Gazella subgutturosa (4.55 %), Canis lupus (27.27 %), Pseudois nayaur (23.08 %), and Ovis ammon (4.55 %). The pan-influenza competitive ELISA results closely corresponded to the cumulative prevalence of AIV exposure as measured by subtype-specific HI assays, suggesting that H7 and H9 AIV subtypes predominate in the wildlife species evaluated. These data provide evidence of prior infection with H7 and H9 AIVs in non-avian wildlife in Xinjiang, China.

Retrospective genomic analysis of the first Lumpy skin disease virus outbreak in China (2019).

Lumpy skin disease caused by Lumpy skin disease virus (LSDV) is a severe systemic disease affecting cattle and other ruminants. Lumpy skin disease was first reported in northwest China in August 2019 and has severely threatened the cattle breeding industry in China. However, there have been limited genomic studies of LSDV from the first outbreak and its subsequent epidemics. This study aims to characterize the comparative genomic evolution of the LSDV strain from the first outbreak in China. The etiological agent was isolated in a Madin-Darby bovine kidney cell culture and subsequently identified by PCR and Sanger sequencing of six selected genes. The genome sequence was determined using Illumina sequencing and analyzed through genome alignment and phylogenetic tree. The results showed that all six genes were successfully amplified and genetically clustered into LSDV. The virus presented the highest homology to strain China/GD01/2020, which shared 100% identities among 150 open reading frames (ORFs), and 97.1-99.7% identities among additional 6 ORFs. Bayesian inference tree analysis revealed that the virus shared a common ancestor with LSDV strains from China and Vietnam. The study provides an additional genomic data for LSDV tracking and control in China and neighboring countries.

Ultrasensitive electrochemiluminescence detection of Staphylococcus aureus via enzyme-free branched DNA signal amplification probe.

As a main cause of foodborne diseases, pathogenic bacteria have threatened the health and well-being of human communities. There is a need of fastness, accuracy and sensitivity in the method of detecting pathogenic bacteria. Classical signal amplification assays usually employ enzymes as biocatalysts to generate amplified signals, but the strict experimental conditions and complicated instruments restrict their application. In this work, we demonstrated an enzyme-free branched DNA (bDNA)-based signal amplification electrochemiluminescence (ECL) assay for ultrasensitive detection of pathogenic bacteria. Firstly, the capture probes and the amplification probes group were carefully designed by our research group. The detecting ECL signal of Staphylococcus aureus (S. aureus) was amplified by bDNA technique through the layer-by-layer signal amplification. The sensitivity was greatly improved by the use of multiple Ru(bpy)32+ (TBR)-labeled ECL probes. Secondly, the whole process of the detection was carried out in the absence of enzyme, without the need to control the reaction conditions strictly. Thirdly, the designed amplification probes group could be used for the analysis of other pathogenic bacteria, virus, tumor markers, biomarkers, etc. For the detection of S. aureus, the limit of detection (LOD) of the method was 2 pM for standard DNA, with the linear range from 20 pM to 100 nM. Last but not least, the LOD of the S. aureus asymmetric PCR products was 5 pM, with the linear range from 10 pM to 50 nM. The sensitivity was 1-2 orders in magnitude higher than that of the common detection assays.

Estrogen is a critical determinant that specifies the duration of the window of uterine receptivity for implantation.

Many underlying causes of human infertility have been overcome by using in vitro fertilization (IVF) and embryo transfer (ET) techniques. Nevertheless, implantation rates in IVF programs remain low despite the transfer of apparently healthy embryos. This suggests that there are problems with the differentiation of the uterus to the receptive state in response to the ovarian hormones estrogen and progesterone. The molecular basis of this receptive state when the uterine environment is conducive to blastocyst acceptance and implantation remains poorly understood. Normally, the "window" of uterine receptivity lasts for a limited time. Using ETs and the progesterone-treated delayed-implantation model in mice, we demonstrate here that levels of estrogen within a very narrow range determine the duration of the window of uterine receptivity. Although estrogen at different physiological concentrations can initiate implantation, we find that the window of uterine receptivity remains open for an extended period at lower estrogen levels but rapidly closes at higher levels. The uterine refractoriness that follows the receptive state at high estrogen levels is accompanied by aberrant uterine expression of implantation-related genes. These results suggest that careful regulation of estrogen levels is one of the important factors for improvement of female fertility in IVFET programs.

Rescue of female infertility from the loss of cyclooxygenase-2 by compensatory up-regulation of cyclooxygenase-1 is a function of genetic makeup.

Cyclooxygenase-2 (COX-2), an inducible rate-limiting enzyme in prostaglandin biosynthesis, is implicated in various physiological and pathological processes including female fertility, renal function, angiogenesis, inflammation, and tumorigenesis. We showed previously that targeted deletion of Ptgs2 encoding COX-2, but not Ptgs1 encoding COX-1, in C57BL/6J/129 mice produces complete female infertility resulting from multiple reproductive failures spanning ovulation, fertilization, and implantation. Here we show that Ptgs2 null mice on a CD1 background have dramatically improved female fertility including ovulation, fertilization, and implantation, giving rise to live births. We provide evidence that this improved fertility in CD1 Ptgs2 null mice is the result of a compensatory up-regulation of Ptgs1 which does not occur in C57BL/6J/129 mice missing Ptgs2. These results clearly demonstrate for the first time that COX-1 can replace specific functions of COX-2 in vivo in the context of genetic disparity. In light of this finding, the therapeutic use and efficacy of COX-2-specific inhibitors among human populations without regard for genetic and ethnic diversities should be revisited.

Cyclooxygenase-2 differentially directs uterine angiogenesis during implantation in mice.

Increased vascular permeability and angiogenesis at the site of blastocyst apposition in the uterus are two hallmarks of the implantation process. The present investigation shows that although the proangiogenic vascular endothelial growth factor (VEGF) and its receptor, Flk-1, are primarily important for uterine vascular permeability and angiogenesis prior to and during the attachment phase of the implantation process, VEGF in complementation with the angiopoietins and their receptor, Tie-2, directs angiogenesis during decidualization following implantation. Mice with null mutation for the gene encoding cyclooxygenase-2 (COX-2), a rate-limiting enzyme in prostaglandin (PG) biosynthesis, show implantation and decidualization failure. Using reporter and mutant mice, we show here that COX-2-derived prostaglandins (PGs) are important for uterine vascular permeability and angiogenesis during implantation and decidualization, suggesting that one cause of the failure of these latter processes in Cox-2-/- mice is the deregulated vascular events in the absence of COX-2. The attenuation of uterine angiogenesis in these mice is primarily due to defective VEGF signaling and not due to the defective angiopoietin system.