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Chronic stress has become a major problem that endangers people’s physical and mental health. Studies have shown that chronic stress impairs female reproduction. However, the related mechanism is not fully understood. P2X7 receptor (P2X7R) is involved in a variety of pathological changes induced by chronic stress. Whether P2X7R is involved in the effect of chronic stress on female reproduction has not been studied. In this study, we established a chronic restraint stress mouse model and chronic cold stress mouse model. We found that the number of corpora lutea was significantly reduced in the two chronic stress models. The number of corpora lutea indirectly reflects the ovulation, suggesting that chronic stress influences ovulation. P2X7R expression was significantly increased in ovaries of the two chronic stress models. A superovulation experiment showed that P2X7R inhibitor A-438079 HCL partially rescued the ovulation rate of the two chronic stress models. Further studies showed that activation of P2X7R signaling inhibited the cumulus expansion and promoted the expression of NPPC in granulosa cells, one key negative factor of cumulus expansion. Moreover, sirius red staining showed that the ovarian fibrosis was increased in the two chronic stress models. For the fibrosis-related factors, TGF-β1 was increased and MMP2 was decreased. In vitro studies also showed that activation of P2X7R signaling upregulated the expression of TGF-β1 and downregulated the expression of MMP2 in granulosa cells. In conclusion, P2X7R expression was increased in the ovaries of the chronic restraint-stress and chronic cold-stress mouse models. Activation of P2X7R signaling promoted NPPC expression and cumulus expansion disorder, which contributed to the abnormal ovulation of the chronic stress model. Activation of P2X7R signaling is also associated with the ovarian fibrosis changes in the chronic stress model.

Grazing by local livestock is the traditional human practice in Qinghai-Tibetan Plateau grassland, and moderate intensity grazing can maintain high productivity and diversity of alpine grassland. Grazing ecosystems are often nitrogen-limited, but N2-fixing communities in response to yak grazing and Tibetan sheep grazing in Qinghai-Tibetan Plateau grassland have remained underexplored. In this study, we applied quantitative PCR quantitation and MiSeq sequencing of nifH under yak grazing and Tibetan grazing through a manipulated grazing experiment on an alpine grassland. The results showed that the grazing treatments significantly increased the soil ammonium nitrogen (AN) and total phosphorus (TP), but reduced the diazotrophs abundance. Compared with no grazing treatment, the composition of diazotrophs could be maximally maintained when the ratio of yak and Tibetan sheep were 1:2. The foraging strategies of grazing livestock reduced the legumes biomass, and thus reduced the diazotrophs abundance. Data analysis suggested that the direct key factors in regulating diazotrophs are AN and TP, and the changes of these two soil chemical properties were affected by the dung and urine of herbivore assemblages. Overall, these results indicated that the mixed grazing with a ratio of yak to Tibetan sheep as 1:2 can stabilize the soil diazotrophsic community, suggesting that MG12 are more reasonable grazing regimes in this region.

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) in foods has been associated with severe infections in humans and animals worldwide. In the present study, the molecular characteristics of livestock-associated MRSA (LA-MRSA) and human-associated MRSA (hMRSA) isolates obtained in China, as well as MRSA isolates obtained from raw milk in 2018, were investigated. In total, 343 (20.38%; 343/1,683) S. aureus isolates were obtained from 1,683 raw milk samples from 100 dairy farms in 11 provinces across China. Among these, 49 (2.91%; 49/1,683) were mecA-positive MRSA. All LA-MRSA isolates were resistant to penicillin and highly resistant to erythromycin, sulfisoxazole, and clindamycin. Bioinformatic analysis the 49 genomes of LA-MRSA and 71 previously published hMRSA genomes isolated from Chinese individuals in 2018 indicated that blaZ, erm, ant(6)-Ia, aph(3′)-III, tet(K), cat, and aph(2″)-Ia were more prevalent in MRSA from raw milk (P

As one of the most widespread renewable energy technologies, photovoltaic power generation system provides great environmental and economic benefits. Achieving precise prediction for the PV power generation will greatly improve the quality of electric energy and enhance the stability of power system operation. Inspired by the powerful ability of fuzzy logic to deal with uncertainty and the superiority of machine learning to handle the time series prediction, a hybrid ensembled model consisted of the Double-Input-Fuzzy-Modules (DIFM) and Extreme Learning Machine is proposed in this paper. Firstly, the PV power generation data is taken as the input of each DIFM in order to efficiently handle the uncertainties. Then, the outputs of each DIFM are used as the input of ELM. Moreover, the least square estimation is applied to train the parameters of the hybrid ensembled model to further enhance the predict precision. Finally, the proposed hybrid ensembled model is applied to achieve a real-world PV power generation forecasting. The cases study and comparisons results indicate that the proposed hybrid ensembled model outperforms other methods, in terms of the mean absolute error, the root means square error and the coefficient of determination.

Three different herbivore grazing assemblages, namely, yak grazing (YG), Tibetan sheep grazing (SG) and yak and Tibetan sheep co-grazing (MG), are practiced in alpine meadows on the Qinghai-Tibetan Plateau (QTP), but the effects of the different herbivore assemblages on soil microbes are relatively unknown. The microbial community plays an important role in the functional stability of alpine grassland ecosystems. Therefore, it is important to understand how the microbial community structure of grassland ecosystems changes under different herbivore grazing assemblages to ensure their sustainable development. To fill this gap, a field study was carried out to investigate the effects of YG, SG, and MG on plant communities, soil physico-chemical properties and microbial communities under moderate grazing intensity in alpine meadows. Grazing increased the β-diversity of the bacteria community and decreased the β-diversity of the fungal community. The herbivore assemblage affected the microbial community diversity, but not the plant community diversity. Total phosphorus, soil bulk density, root biomass, and plant α-diversity were correlated with both the bacterial and fungal community composition, available phosphorus and soil moisture were correlated only with the bacterial community composition, while available potassium and above-ground net primary production (ANPP) were correlated only with the fungal community composition. Soil available nitrogen, soil available phosphorus and soil bulk density were highest in SG, while ANPP was highest in MG. It was concluded that MG can improve ANPP and stabilize the soil microbial community, suggesting that MG is an effective method for sustainable use and conservation of alpine meadows on the QTP.

Polycystic ovary syndrome (PCOS) is an endocrine disorder and metabolic syndrome. Ovarian fibrosis pathological change in PCOS has gradually attracted people’s attention. In this study, we constructed a PCOS mouse model through the use of dehydroepiandrosterone. Sirius red staining showed that the ovarian tissues in PCOS mice had obvious fibrosis. Prolyl oligopeptidase (POP) is a serine protease and N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is its catalytic product. Studies show that abnormal expression and activity of POP and Ac-SDKP are closely related to tissue fibrosis. It was found that the expression of POP and Ac-SDKP was decreased in the ovaries of PCOS mice. Further studies showed that POP and Ac-SDKP promoted the expression of matrix metalloproteinases 2 (MMP-2) expression and decreased the expression of transforming growth factor beta 1 (TGF-β1) in granulosa cells. Hyperandrogenemia is a typical symptom of PCOS. We found that testosterone induced the low expression of POP and MMP2 and high expression of TGF-β1 in granulosa cells. POP overexpression and Ac-SDKP treatment inhibited the effect of testosterone on TGF-β1 and MMP2 in vitro and inhibited ovarian fibrosis in the PCOS mouse model. In conclusion, PCOS ovarian tissue showed obvious fibrosis. Low expression of POP and Ac-SDKP and changes in fibrotic factors contribute to the ovarian pathological fibrosis induced by androgen.

ABSTRACT: During poultry slaughter, cross-contamination of chicken carcasses with microorganisms (including drug-resistant bacteria) can occur because of incomplete disinfection during the pre-cooling process, and surface contact with contaminated tools and equipment. The use of disinfectants is the most common way to reduce the risk of cross-contamination and bacterial spread, as they can effectively reduce the number of bacteria. We developed a disinfectant consisting of organic acids and sodium dodecyl sulfate (SDS) and tested its bactericidal effects at different concentrations against Salmonella and Campylobacter. The main effective components in the disinfectant were citric acid, lactic acid, and SDS, and together they exerted a synergistic bactericidal effect. The bactericidal efficacy of the disinfectant increased with increasing concentrations of the 3 active ingredients. To reach a 100% reduction rate during a 15-s treatment in vitro, for Salmonella, the lowest concentrations of citric acid, lactic acid, and SDS were 0.06, 0.08, and 0.02%, respectively; and for Campylobacter, the lowest concentrations were 0.02, 0.025, and 0.0125%, respectively. The disinfectant remained effective in presence of interfering substances (e.g., 15% fetal bovine serum). Further experiments showed that the disinfectant inactivated sensitive bacteria as well as 23 drug-resistant strains of Salmonella and Campylobacter. Treatment with the disinfectant for 15 s decreased the concentrations of all tested strains by more than 4.7 log colony forming units per mL, and the reduction rate was as high as 100%. In on-site disinfection tests in chicken slaughterhouses, the disinfectant significantly reduced the number of pathogenic bacteria on carcasses during the pre-cooling process, and on tools (such as knives and gloves) during the segmentation process. Thus, this disinfectant has potential uses in preventing cross-contamination of food-borne pathogens (including resistant bacteria) in slaughterhouses.

Abstract Background Poa L. is a large genus of grass in Gramineae, among which P. pratensis is widely cultivated as turf and forage. Satellite DNA is the main components of the plant genome. Information of satellites will helpful for dissection the genome composition and definition of the phylogeny relationship of these species. However, the knowledge about the satellites in genus Poa is still limited. Results Four satellite DNAs were identified using the Repeat Explorer pipeline in HiSeq Illumina reads from diploid plants in P. malaca (2n = 26). Two satellites showed high similarity with the previously identified PpTr-1 and PpTr-3, whereas two others are newly identified with the monomer of 326 bp (Poa-326) and 353 bp (Poa-353) respectively. The clone DNAs of PpTr-1 and PpTr-3, and oligonucleotides designed representing satellites Poa-326 and Poa-353 were probed to test on chromosomes across 13 Poa speceis with different polyploidy level by fluorescent in situ hybridization (FISH). PpTr-1, PpTr-3, and Poa-362 were stably positioned in the subtelomeric regions in nearly all species with the variation of hybridization sites number. However, Poa-353 showed different FISH patterns of multiple regions with the variation of hybridization intensity and distribution sites across species. In addition, 5S rDNA and 45S rDNA were used to characterize the genome of the Poa species. Four rDNA FISH patterns were revealed in the tested species. Conclusion Four identified satellite were high conservable across Poa species. Genome distribution of these satellites can be characterized by FISH. The variation of satellite DNAs and rDNA chromosomal distributions between species provide useful information for phylogenetic analysis in genus Poa.

Methicillin-resistant staphylococcus aureus (MRSA) is a major pathogen responsible for human hospital and community-onset diseases and severe invasive livestock infections. Rapid detection of MRSA is essential to control the spread of MRSA. Conventional identification methods and antibacterial susceptibility tests of MRSA are time-consuming. The commonly used qPCR assay also has the disadvantages of being complicated and expensive, restricting its application in resource-limited clinical laboratories. Here, a real-time fluorescent recombinase-assisted amplification (RAA) assay targeting the most conserved regions within the mecA gene of MRSA was developed and evaluated to detect MRSA. The detection limit of this assay was determined to be 10 copies/reaction of positive plasmids. The established RAA assay showed high specificity for MRSA detection without cross-reactivities with other clinically relevant bacteria. The diagnostic performance of real-time RAA was evaluated using 67 clinical S. aureus isolates from dairy farms, which were detected in parallel using the TaqMan probe qPCR assay. The results showed that 56 and 54 samples tested positive for MRSA by RAA and qPCR, respectively. The overall agreement between both assays was 97.01% (65/67), with a kappa value of 0.9517 (p < 0.001). Further linear regression analysis demonstrated that the detection results between the two assays were significantly correlated (R2 = 0.9012, p < 0.0001), indicating that this RAA assay possesses similar detection performance to the qPCR assay. In conclusion, our newly established RAA assay is a time-saving and convenient diagnostic tool suitable for MRSA detection and screening.

Human immunodeficiency virus envelope glycoprotein 120 (gp120) leads to hyperalgesia. Long non-coding RNAs are characterized by the lack of a protein-coding sequence and may contribute to the development and maintenance of inflammatory and neuroinflammatory pain. Rats with neuroinflammatory pain were established by gp120 treatment, which is featured by intensified pain behaviors. Long non-coding RNA uc.48+ was increased in the dorsal root ganglia of gp120-treated rats, and small interfering RNA that targets uc.48+ markedly alleviated hyperalgesia in gp120-treated rats. Notably, uc.48+ overexpression increased P2Y12 expression in control rats dorsal root ganglia and induced hyperalgesia. Uc.48+ small interfering RNA inhibited P2Y12 expression in gp120-treated rats. Uc.48+ potentiated P2Y12 receptor functions in the neurons and heterologous cells. Therefore, uc.48+ siRNA treatment reduced the upregulation of P2Y12 expression and function in DRG neurons, and, hence, alleviated hyperalgesia in gp120-treated rats.

Muscle regeneration depends on self-renewal of muscle stem cells but how this is regulated on aging is unclear. Here, the authors identify Slug as regulating p16 Ink4a in quiescent muscle stem cells, and when Slug expression reduces in aged stem cells, p16Ink4a accumulates, causing regenerative defects.

Quercetin has potential pharmacological values in various carcinomas including oral squamous cell carcinoma (OSCC). Moreover, the anti-tumor effect of quercetin is correlated with WNT/β-catenin pathway and miRNA dysregulation. In the present study, we aimed to further investigate whether quercetin can exert its anti-tumor function by regulating miR-22 together with miR-22 downstream pathway WNT1/β-catenin in OSCC. The results of Cell Counting Kit-8 (CCK-8) and flow cytometry analyses showed that quercetin treatment and miR-22 overexpression resulted in the reduction of cell viability and the increase of cell apoptotic rate in OSCC. WNT1 was a target of miR-22, which was confirmed by bioinformatics, luciferase reporter and RNA immunoprecipitation (RIP) assays. RT-qPCR assay showed that quercetin promoted miR-22 expression and suppressed WNT1 and β-catenin expression in OSCC cells, whereas this effect was abrogated by miR-22 inhibitor. Moreover, miR-22 depletion weakened quercetin-mediated viability inhibition and apoptosis increase in OSCC cells. Quercetin inhibited the growth of OSCC xenograft tumors by inducing miR-22 expression and repressing WNT1/β-catenin pathway in vivo. Taken together, quercetin hampered OSCC tumorigenesis by regulating miR-22/WNT1/β-catenin pathway in OSCC, providing a deep insight into the molecular targets of quercetin in the treatment of OSCC. Keywords: Quercetin, microRNA-22, WNT1/β-catenin pathway, Oral squamous cell carcinoma

ABSTRACTThe recent emergence of plasmid-mediated tigecycline resistance genes, tet(X3) and tet(X4), in animals and humans in China would pose a foreseeable threat to public health. To illustrate this paradigm shift in tigecycline resistance, here, covering the period 2008-2018, we retrospectively analysed a national strain collection of Escherichia coli (n = 2254), obtained from chickens and pigs, in six representative provinces of China. The gene tet(X4) was identified in five pig isolates collected in 2016 and 2018 from the provinces of Sichuan (3/15, 2018), Henan (1/25, 2018) and Guangdong (1/28, 2016), but not in the isolates prior to 2016. None of the isolates was detected harbouring tet(X3). All tet(X4)-positive E. coli exhibited high levels of tigecycline resistance (MICs, 16-64 mg/L), and two were confirmed as colistin resistant, harbouring chromosome-borne mcr-1 gene. The gene tet(X4) was detected on a plasmid in all five isolates, whereas a co-location of tet(X4) on the chromosome of one isolate was observed. Diverse host strains and novel plasmids related to the tet(X4) gene were observed. Our timely findings of the recent emergence of tet(X4) gene in food animal support the rapid surveillance and eradication of this gene before it is established.

Human activity, combined with climate change, has led to severe degradation of natural grasslands all over the world. As such, artificial grasslands created with species suited to the geographical area present a popular alternative. However, species that are both native and suitable for creating artificial grasslands are rare, especially in the Qinghai-Tibetan Plateau of China. In this study, we performed two experiments respectively with varied temperature and water potential, to evaluate the cold and drought resistance of 13 local Poa L. species in the Qinghai Province of China. Within the range of 0–20 °C, our results showed that increase in temperature increased the germination indices of all 13 species, with some species germinating even at 5 °C, demonstrating relatively cold-resistant species in our study. Similarly, decrease in moisture levels decreased the germination percentages of all species in this experiment, with several species germinating at even −1.2 MPa, which were considered as relatively higher drought-resistant species in this study. Among the 13 species, P. malaca, P. paucifolia, and P. sphondylodes exhibited relatively strong cold-resistance, while P. poophagorum, P. pratensis var. anceps Gaud. cv. Qinghai, and P. pratensis cv. Qinghai showed relatively high drought resistance. Overall, the results indicated that all the 13 native grass species evaluated in the study were suitable for planting and cultivation on the Qinghai-Tibetan Plateau. This study could provide insights into identifying suitable species for plant breeding in Qinghai-Tibetan plateau and establish a protocol for germplasm evaluation of local Poa L. species. Further, these findings can help evaluate and identify more native species suited for the restoration of degraded grasslands, especially under the scenario of climate change.

LncPrep + 96kb is a novel long non-coding RNA expressed in murine granulosa cells with two transcripts that are 2.2 and 2.8 kb in length. However, the potential roles of lncPrep + 96kb in granulosa cells remain poorly understood. In this study, we investigated the effect of the lncPrep + 96kb 2.2 kb transcript on granulosa cells through the overexpression and knockdown of lncPrep + 96kb 2.2 kb. We found that lncPrep + 96kb 2.2 kb inhibited aromatase expression and estradiol production. Endothelial differentiation-related factor 1 (EDF1) is an evolutionarily conserved transcriptional coactivator. We found that EDF1 knockdown inhibited aromatase expression and estradiol production. The RNA immunoprecipitation results also showed that lncPrep + 96kb 2.2 kb can bind to EDF1 and that overexpression of lncPrep + 96kb 2.2 kb induced the translocation of EDF1 from the nucleus to the cytoplasm. The CatRAPID signature revealed that the 1,979–2,077 and 603–690 nucleotide positions in lncPrep + 96kb 2.2 kb were potential binding sites for EDF1. We found that mutating the 1,979–2,077 site rescued the effects of lncPrep + 96kb 2.2 kb on aromatase expression and estradiol production. In conclusion, we are the first to report that specific expression of lncPrep + 96kb 2.2 kb in granulosa cells inhibits the production of estradiol by influencing the localization of EDF1 in granulosa cells. The 1,979–2,077 site of lncPrep + 96kb 2.2 kb contributes to the ability to bind to EDF1.

Background Stoichiometric relations drive powerful constraints in several fundamental ecosystem processes. However, limited studies have been conducted on the ecological stoichiometry of plants after the change of community composition induced by Stellera chamaejasme removal in alpine grassland in the Qinghai–Tibetan Plateau. Methods We investigated the effects of S. chamaejasme removal on ecological stoichiometry by estimating the C:N:P stoichiometry in species, functional group and community levels of the ecosystem. The interactions between different species, functional groups and correlation with soil nutrient, responding to S. chamaejasme removal were also analyzed. Results For the plants that became dominant after S. chamaejasme removal (SR), N content decreased and their C:N increased. S. chamaejasme removal significantly affected the nutrient stoichiometry of different functional groups. Specifically, Gramineae in the SR sites had decreased N content and N:P, and increased C:N; however, forbs had increased N content, C:P and N:P and decreased P content and C:N. At the community level, N content was lower and C:N higher in SR communities compared to CK. The N content of the plant community was positively correlated with soil total N content. S. chamaejasme removal could change the nutrient balance from species level, to functional group level, and to community level. Thus, supplementary measures might be cooperated with S. chamaejasme removal for the recovery of S. chamaejasme-dominated degraded grassland. These results provide insight into the role of S. chamaejasme in ecological protection and conservation, and the conclusions from this study could be used to develop effective and sustainable measures for S. chamaejasme control in the Qinghai–Tibetan Plateau.

This study aimed to obtain a simple and efficient soil extraction method as an exhaustive and systematic technology guideline for the determination of potentially available phosphorus (P), potassium (K), magnesium (Mg), iron (Fe), copper (Cu), manganese (Mn) and zinc (Zn) in various grassland soils covering different regions. In this study, 25 soil samples from 5 different grassland types of China were collected to measure the amounts of potentially available nutrients and to compare the results of Mehlich 3, the ammonium bicarbonate-diethylenetriaminepentaacetic acid (AB-DTPA) and other four methods which are widely used in China (i.e. Olsen, Bray 1, 1 mol/L NH4OAc, CaCl2-DTPA, called conventional methods). The results showed that the amounts of potentially available nutrients extracted by different methods were significantly different. Moreover, a positively significant correlation was obtained between the amounts of potentially available nutrients extracted by conventional methods and those extracted by Mehlich 3 and AB-DTPA. However, for Mg and Mn, the results from AB-DTPA appeared to have a stronger relationship with the results from the conventional method than with the results from Mehlich 3. These differences might be caused by the effect of soil pH. Our results suggested that both Mehlich 3 and AB-DTPA can be effectively used to measure nutrients availability in grassland soils, while the AB-DTPA will be more recommended to measure the amounts of potentially available nutrients in alkaline soils.

Abstract Over these years, more and more sex cord-stromal tumors have been reported. Granulosa cell tumor (GCT) is a rare tumor in ovaries, accounts for 2% to 5% of ovarian cancers. The main different feature of GCTs from other ovarian cancers is that GCTs can lead to abnormally secreted hormones (estrogen, inhibin and Müllerian inhibiting substance). The GCT is divided into two categories according to the age of patients, namely AGCT (adult granulosa cell tumor) and JGCT (Juvenile granulosa cell tumor). AGCT patients accounts for 95%. Although the pathogenesis is not clear, FOXL2 (Forkhead box L2) mutation was considered as the most critical factor in AGCT development. The current treatment is dominated by surgery. Target therapy remains in the adjuvant therapy stage, such as hormone therapy. During these years, other pathogenic factors were also explored, such as PI3K/AKT (phosphatidylinositol-3-kinase; serine/threonine kinase), TGF-β (Transforming growth factor beta) signaling pathway, Notch signaling pathway, GATA4 and VEGF (vascular endothelial growth factor). These factors and signaling pathway play important roles in GCT cell proliferation, apoptosis, or angiogenesis. The purpose of this review is to summarize the possible pathogenic factors and signaling pathways, which may shed lights on developing potential therapeutic targets for GCT.

In the present work, the influence of filling rate on macrosegregation in a 40-Metric Ton (MT) ingot of a high-strength low-carbon steel was studied using finite element (FE) simulation. The modelling of the filling and solidification processes were realized with a two-phase (liquid-solid) multiscale 3D model. The liquid flow induced by the pouring jet, the thermosolutal convection, and the thermomechanical deformation of the solid phase were taken into consideration. Two filling rates were examined, representing the upper and lower manufacturing limits for casting of large size ingots made of high strength steels for applications in energy and transportation industries. The evolution of solute transport, as well as its associated phenomena throughout the filling and cooling stages, were also investigated. It was found that increasing the filling rate reduced macrosegregation intensity in the upper section, along the centerline and in the mid-radius regions of the ingot. The results were analyzed in the framework of heat and mass transfer theories, liquid flow dynamics, and macrosegregation formation mechanisms.

This study provides a chloroplast genome of Poa pratensis L. cv. Qinghai. The complete genome is 135,606 bp in length with a G + C content of 38.28%, which contains 32 tRNA genes, and 8 rRNA genes. The phylogenetic analysis indicated that P. pratensis L. cv. Qinghai is closely related to Festuca arundinacea cultivar KY-31. These results contribute to explore the genetic evidence for adaptation to the Qinghai-Tibet Plateau.

This study provides a chloroplast genome of Poa poophagorum. The complete cp genome was135,664 bp in length with typical quadripartite structure, containing a pair of inverted repeats (IR) of 21,552 bp each, a large single-copy (LSC) region of 79,790 bp, and a small single-copy (SSC) region of 12,770 bp. The overall G + C content of the cp genome was 38.30%, which encompassed 119 genes including 79 protein-coding genes,8 rRNA genes, and 32 tRNA genes. The phylogenetic analysis indicated that P. poophagorum was closely related to Festuca arundinacea cultivar KY-31in Gramineae. This study would contribute to enrich the Poa L. cp genome resource and promote biological research.

The objectives of this study were to investigate the prevalence and fluoroquinolone resistant Salmonella isolated from an integrated broiler chicken supply chain and their molecular characterization. In total, 73 Salmonella isolates were recovered from a broiler chicken supply chain in Shanghai. Salmonella isolates were tested for susceptibility to 11 antimicrobial agents using the broth dilution method and were characterized using pulsed-field gel electrophoresis (PFGE). Then, the Salmonella isolates were examined for mutations in quinolone resistance-determining region (QRDR) of gyrA, gyrB, parC, and parE, and were screened for plasmid-mediated quinolone resistance (PMQR) genes. Lastly, we sequenced the plasmids carrying qnrS1 in six Salmonella isolates from three sources (two isolated per source). Among 73 Salmonella isolates, 45 isolates were identified as S. Indiana, 24 were S. Schwarzengrund, 2 were S. Enteritidis, and 2 were S. Stanleyville. In addition, high rates of resistance were detected for nalidixic acid (41.1%) and ciprofloxacin (37.0%), while resistance to other test agents was diverse (2.0–100%). S. Indiana and S. Schwarzengrund isolates from different sources exhibited the same PFGE pattern, suggesting that the Salmonella isolates possessed high potential to spread along the broiler chicken supply chain. gyrA and parC exhibited frequent missense mutations. Moreover, qnrS1 was the most prevalent PMQR gene in the 73 Salmonella isolates, and it was found about a new hybrid plasmid. This study concludes a high prevalence of fluoroquinolone resistant Salmonella in chicken supply chain, threatening the treatment of Salmonella foodborne diseases. In particular, the emergence of a new hybrid plasmid carrying qnrS1 indicates that the recombination of plasmid carrying resistance gene might be a potential risk factor for the prevention and control strategies of drug resistance.

The extracellular adenosine triphosphate (ATP) regulates different cellular functions through activating purinergic receptors as a signalling molecule or neurotransmitter. P2X7 is highly expressed in murine small luteal cells. In this study, murine luteal cells were cultured in vitro and treated with P2X7 agonists – ATP and 2′(3′)-O-(4-benzoyl-benzoyl)-adenosine 50-triphosphate (BzATP) and with P2X7 antagonist – brilliant blue G (BBG). We found that ATP and BzATP increased the production of progesterone and had no influence on the production of estradiol. BBG reversed the effect of BzATP and ATP. Further studies demonstrated that ATP and BzATP promoted the expression of CYP11A. These results revealed that P2X7 receptor activation is involved in the steroid synthesis in corpus luteum.

The impact of microsegregation models on thermophysical properties and solidification behaviors of a high strength steel was investigated. The examined microsegregation models include the classical equilibrium Lever rule, the extreme non-equilibrium Scheil-Gulliver, as well as other treatments in the intermediate regime proposed by Brody and Flemings, Clyne and Kurz, Kobayashi and Ohnaka. Based on the comparative analyses performed on three representative regions with varied secondary dendrite arm spacing sizes, the classical equilibrium Lever rule and non-equilibrium Scheil scheme were employed to determine the thermophysical features of the studied steel, using the experimentally verified models from literature. The evaluated thermophysical properties include effective thermal conductivity, specific heat capacity and density. The calculated thermophysical data were used for three-dimensional simulation of the casting and solidification process of a 40 metric ton steel ingot, using FEM code Thercast®. The simulations captured the full filling, the thermo-mechanical phenomena and macro-scale solute transport in the cast ingot. The results demonstrated that Lever rule turned out to be the most reasonable depiction of the physical behavior of steel in study in large-size cast ingot and appropriate for the relevant macrosegregation simulation study. The determination of the model was validated using the experimentally measured top cavity dimension, the thermal profiles on the mold outside surface by means of thermocouples, and the carbon distribution patterns via mass spectrometer analysis.

Myocardial ischemia (MI) is one of the major causes of death in cardiac diseases. Purinergic signaling is involved in bidirectional neuronal-glial communication in the primary sensory ganglia. The sensory neuritis of cardiac afferent neurons in cervical dorsal root ganglion (cDRG) interacts with cardiac sympathetic efferent postganglionic neurons, forming feedback loops. The P2Y12 receptor is expressed in satellite glial cells (SGCs) of DRG. Baicalin is a major active ingredient extracted from natural herbal medicines, which has anti-inflammatory and strong anti-oxidation properties. In this study we investigated the effect of baicalin on P2Y12 receptor in the cervical DRG SGC-mediated sympathoexcitatory reflex, which is increased during MI. The results showed that the expression of P2Y12 receptor mRNA and protein in DRG, and the co-localization values of P2Y12 receptor and glial fibrillary acidic protein (GFAP) in cDRG SGCs were increased after MI. The activated SGCs increased IL-1β protein expression and elevated Akt phosphorylation in cDRG. Baicalin treatment inhibited the upregulation of the P2Y12 receptor, GFAP protein and Akt phosphorylation in cDRG neurons/SGCs. The stellate ganglia (SG) affect cardiac sympathetic activity. Baicalin treatment also decreased the upregulation of the P2Y12 receptor, GFAP protein in the SG. The P2Y12 agonist, 2Me-SADP, increased [Ca2+]i in HEK293 cells transfected with the P2Y12 receptor plasmid and SGCs in cDRG. These results indicate that application of baicalin alleviates pathologic sympathetic activity induced by MI via inhibition of afferents in the cDRG.

Aim: In this study, we investigated whether andrographolide (Andro) can alleviate neuropathic pain induced by HIV gp120 plus ddC treatment and the mechanism of its action.Methods: The paw withdrawal threshold and the paw withdrawal latency were observed to assess pain behaviors in all groups of the rats, including control group, control combined with Andro treatment group, sham group, gp120 combined with ddC treatment group, gp120 plus ddC combined with A438079 treatment group, and gp120 plus ddC combined with Andro treatment by intrathecally injecting at a dose of 25 μg/20 μl group. The protein expression levels of the P2X7 receptor, tumor necrosis factor-α-receptor (TNFα-R), interleukin-1β (IL-1β), IL-10, phospho-extracellular regulated protein kinases (ERK) (p-ERK) in the L4–L6 dorsal root ganglia (DRG) were measured by western blotting. Real-time quantitative polymerase chain reaction was used to test the mRNA expression level of the P2X7 receptor. Double-labeling immunofluorescence was used to identify the co-localization of the P2X7 receptor with glial fibrillary acidic protein (GFAP) in DRG. Molecular docking was performed to identify whether the Andro interacted perfectly with the rat P2X7 (rP2X7) receptor.Results: Andro attenuated the mechanical and thermal hyperalgesia in gp120+ddC-treated rats and down-regulated the P2X7 receptor mRNA and protein expression in the L4–L6 DRGs of gp120+ddC-treated rats. Additionally, Andro simultaneously decreased the expression of TNFα-R and IL-1β protein, increased the expression of IL-10 protein in L4–L6 DRGs, and inhibited the activation of ERK signaling pathways. Moreover, Andro decreased the co-expression of GFAP and the P2X7 receptor in the SGCs of L4–L6 DRG on 14th day after surgery.Conclusion: Andro decreased the hyperalgesia induced by gp120 plus ddC.

Diabetic peripheral neuropathy results in diabetic neuropathic pain (DNP). Satellite glial cells (SGCs) enwrap the neuronal soma in the dorsal root ganglia (DRG). The purinergic 2 (P2) Y12 receptor is expressed on SGCs in the DRG. SGC activation plays an important role in the pathogenesis of DNP. Curcumin has anti-inflammatory and antioxidant properties. Because curcumin has poor metabolic stability in vivo and low bioavailability, nanoparticle-encapsulated curcumin was used to improve its targeting and bioavailability. In the present study, our aim was to investigate the effects of nanoparticle-encapsulated curcumin on DNP mediated by the P2Y12 receptor on SGCs in the rat DRG. Diabetic peripheral neuropathy increased the expression levels of the P2Y12 receptor on SGCs in the DRG and enhanced mechanical and thermal hyperalgesia in rats with diabetes mellitus (DM). Up-regulation of the P2Y12 receptor in SGCs in the DRG increased the production of pro-inflammatory cytokines. Up-regulation of interleukin-1β (IL-1β) and connexin43 (Cx43) resulted in mechanical and thermal hyperalgesia in rats with DM. The nanoparticle-encapsulated curcumin decreased up-regulated IL-1β and Cx43 expression and reduced levels of phosphorylated-Akt (p-Akt) in the DRG of rats with DM. The up-regulation of P2Y12 on SGCs and the up-regulation of the IL-1β and Cx43 in the DRG indicated the activation of SGCs in the DRG. The nano-curcumin treatment inhibited the activation of SGCs accompanied by its anti-inflammatory effect to decrease the up-regulated CGRP expression in the DRG neurons. Therefore, the nanoparticle-encapsulated curcumin treatment decreased the up-regulation of the P2Y12 receptor on SGCs in the DRG and decreased mechanical and thermal hyperalgesia in rats with DM.

The complete chloroplast genome from Elymus sibiricus Linn, an important forage grass of the Gramineae, is determined in this study. The whole chloroplast genome sequence of E. sibiricus Linn has been characterized by Illumina pair-end sequencing. The circular genome is 135,074 bp long, containing a large single copy region (LSC) of 80,681 bp and a small single copy region (SSC) of 12,767 bp, which are separated by a pair of 20,813 bp inverted repeat regions (IRs). It encodes a total of 142 genes, including 86 protein-coding genes (81 PCG species), 48 tRNA genes (30 tRNA species) and eight ribosomal RNA genes (four rRNA species). The most of gene species occur as a single copy, while 16 gene species occur in double copies. The overall A + T content of is 61.7%, while the corresponding values of the LSC, SSC and IR regions are 63.6%, 67.8% and 56.0%, respectively. Phylogenetic analysis suggested that E. sibiricus Linn was relatively close to Connorochloa tenuis compared with species of other generain Gramineae among the species analyzed. This complete chloroplast genome will provide valuable insight into conservation and exploitation efforts for this species as well as useful resources for studying the Gramineae phylogeny.

The effect of filling velocity on positive macrosegregation in large size steel ingots was studied. Macrosegregation and macro/microstructures were characterized on the hot-tops and a portion of the upper section of two ingots. The measurements revealed that segregation features in the two ingots varied as a function of the alloying elements, and that the severity of positive macrosegregation in the casting body was reduced when the filling rate was increased. It was also found that at the higher filling rate, grain morphologies in the first solidified zones of the ingot changed from columnar to equiaxe, and secondary dendrite arm spacing (SDAS) became slightly smaller in the intermediate and final solidified zones. The experimental findings were analyzed in the framework of diffusion and convection-controlled solidification, as well as liquid metal flow theories. The solute dependence of segregation features was related to the difference in the solid-liquid partition coefficient and diffusion capability of each element in the liquid iron. Calculation of Reynolds numbers (Re) during the filling process, for both ingots, showed that higher filling velocity caused more instable movement of the liquid metal in the initial solidification stage, resulting in the modification of grain morphology, as well as accelerated solidification rate.

Hybrid complexes with N,N′-bis(2-benzimidazolylmethyl)amine and cyclen moieties are novel enzyme mimics and controlled DNA release materials, which could interact with DNA through three models under different conditions. In this paper, the interactions between plasmid DNA and seven different complexes were investigated, and the methods to change the interaction patterns by graphene oxide (GO) or concentrations were also investigated. The cleavage of pUC19 DNA promoted by target complexes were via hydrolytic or oxidative mechanisms at low concentrations ranging from 3.13 × 10−7 to 6.25 × 10−5 mol/L. Dinuclear complexes 2a and 2b can promote the cleavage of plasmid pUC19 DNA to a linear form at pH values below 7.0. Furthermore, binuclear hybrid complexes could condense DNA as nanoparticles above 3.13 × 10−5 mol/L and partly release DNA by graphene oxide with π-π stacking. Meanwhile, the results also reflected that graphene oxide could prevent DNA from breaking down. Cell viability assays showed dinuclear complexes were safe to normal human hepatic cells at relative high concentrations. The present work might help to develop novel strategies for the design and synthesis of DNA controllable releasing agents, which may be applied to gene delivery and also to exploit the new application for GO.

Background • Although the level of medical care has been improved in recent years, the probability of patients contracting pathogens has increased greatly, with a rising incidence of invasive fungal infections. Deep-seated fungi have become common pathogens of nosocomial infections. Objective • This study aims to systematically assess the effectiveness, mortality, survival rate, and adverse reactions (ARs) of high-dose (HD) liposomal amphotericin B (L-AMB) for human diseases. Methods • Ten articles (1661 patients) of randomized controlled trials (RCTs; whether randomized, singleblind, or double-blind) from January 1, 1960, to December 31, 2020, of HD-L-AMB treatment of diseases were retrieved from the PubMed, Embase, Scopus, Web of Science, and Cochrane Central Register of Controlled Trials databases. The primary outcome measure was the overall therapeutic effect, and the secondary outcome measures were mortality, ≥10-week survival, and ARs. Data were meta-analyzed using RevMan 5.3. Results • Ten RCTs involving 1661 patients were included. HD-L-AMB did not show a significant therapeutic advantage in anti-infection treatment. In addition, HD-LAMB treatment of invasive Aspergillus infection led to high mortality and low survival (≥10 weeks, OR = 0.57, 95%CI 0.34–0.94, P = .03). According to subgroup analysis, the incidence of ARs and the incidence of renal dysfunction associated with invasive fungal infection treatment were higher with HD-L-AMB than with regular-dose L-AMB. Conclusion • HD-L-AMB had no obvious advantage for the treatment of diseases and was accompanied by increased mortality, reduced long-term survival, and increased ARs (including renal insufficiency). Therefore, the use of HD-L-AMB to control infections is recommended with caution only when the preferred treatment is contraindicated. [ABSTRACT FROM AUTHOR]