Characterization of Extraintestinal Pathogenic Escherichia coli Strains Causing Canine Pneumonia in China: Antibiotic Resistance, Virulence Genes, and Sequence Typing.

To determine the etiological agents responsible for acute pneumonia in puppies in China, this study utilized bronchoalveolar lavage (BAL) fluid extraction to enable the isolation, culture, biochemical identification, and 16S rRNA PCR amplification of the pathogens. Following preliminary identification, the pathogens underwent analysis for antibiotic resistance phenotypes and resistance genes. Additionally, the study examined the presence of virulence genes, conducted multilocus sequence typing (MLST), and performed whole-genome sequencing (WGS). The findings revealed that all four isolated pathogens were characterized as extraintestinal pathogenic Escherichia coli (ExPEC). The examined ExPEC strains demonstrated resistance to cephalosporins, tetracyclines, and penicillins, while remaining susceptible to aminoglycosides, beta-lactamase inhibitors, carbapenems, chloramphenicols, and sulfonamides. An analysis of virulence genes identified the presence of eight genes, namely CNF-I, fyuA, fimC, papC, ompA, fimH, irp2, and iroN, which are implicated in their invasiveness and potential to inflict tissue damage. The MLST analysis revealed that all ExPEC strains were classified under either sequence type ST131 (Achtman database) or ST43 (Pasteur database). The study further determined that these strains were absent in the kennel's drinking water source, thereby ruling out water contamination as a potential factor in the emergence of ST131-type ExPEC. This study offers a theoretical framework and empirical evidence for elucidating the potential pathogenic mechanisms and clinical therapeutic strategies of ExPEC in the etiology of acute pneumonia in puppies.

Corylifol A suppresses osteoclastogenesis and alleviates ovariectomy-induced bone loss via attenuating ROS production and impairing mitochondrial function.

Osteoporosis is a systemic disease characterized by an imbalance in bone homeostasis, where osteoblasts fail to fully compensate for the bone resorption induced by osteoclasts. Corylifol A, a flavonoid extracted from Fructus psoraleae, has been identified as a potential treatment for this condition. Predictions from network pharmacology and molecular docking studies suggest that Corylifol A exhibits strong binding affinity with NFATc1, Nrf2, PI3K, and AKT1. Empirical evidence from in vivo experiments indicates that Corylifol A significantly mitigates systemic bone loss induced by ovariectomy by suppressing both the generation and activation of osteoclasts. In vitro studies further showed that Corylifol A inhibited the activation of PI3K-AKT and MAPK pathways and calcium channels induced by RANKL in a time gradient manner, and specifically inhibited the phosphorylation of PI3K, AKT, GSK3 ß, ERK, CaMKII, CaMKIV, and Calmodulin. It also diminishes ROS production through Nrf2 activation, leading to a decrease in the expression of key regulators such as NFATcl, C-Fos, Acp5, Mmp9, and CTSK that are involved in osteoclastogenesis. Notably, our RNA-seq analysis suggests that Corylifol A primarily impacts mitochondrial energy metabolism by suppressing oxidative phosphorylation. Collectively, these findings demonstrate that Corylifol A is a novel inhibitor of osteoclastogenesis, offering potential therapeutic applications for diseases associated with excessive bone resorption.

Systemic lupus erythematosus Association between Osteomyelitis: A two-sample Mendelian randomization study in European population.

Background: Systemic lupus erythematosus (SLE) has been correlated with osteomyelitis (OM), yet the underlying causal relationship remains poorly understood. This study aims to investigate the causal association between SLE and OM using Mendelian randomization (MR) analysis. Methods: Genetic instrumental variables (IVs) correlated with SLE were extracted from a comprehensive genome-wide association study (GWAS) summary database (5201 cases and 9066 controls). OM was considered a SLE phenotype, and summary data from the fast GWA data portal were utilized for the analysis. Eligible IVs were extracted following rigorous quality control measures (P < 5 × 10-8, LD r2>0.001, distance 1 Mb, and F > 10). MR analysis was conducted using the Inverse Variance Weighted (IVW), MR-Egger, and Weighted Median (WM) methods after excluding potential confounders. Cochran's Q was applied for heterogeneity test. Pleiotropy was evaluated through MR-Egger intercept, MR-Pleiotropy Residual Sum and Outlier (MR-PRESSO) method, and Leave-one-SNP-out analysis. Result: A total of 40 eligible IVs were included for MR analysis. IVW results demonstrated a positive causal association between SLE and OM (P = 0.049, OR = 1.167). Heterogeneity analysis reveal no significant heterogeneity in the IVW analysis (P = 0.5503). Pleiotropy tests, including MR-PRESSO global test and MR-Egger intercept, indicated no evidence of pleiotropy in our findings (P > 0.05). Additionally, the Leave-one-SNP-out analysis showed no substantial deviations when removing individual SNPs, thus supporting the robustness of our results. Conclusion: This study establishes a genetic causal relationship between SLE and OM, indicating an increased risk of developing OM in individuals with SLE. Therefore, proactive management of SLE is advised to mitigate the risk of developing OM.

Fixators dynamization for delayed union and non-union of femur and tibial fractures: a review of techniques, timing and influence factors.

The optimal balance between mechanical environment and biological factors is crucial for successful bone healing, as they synergistically affect bone development. Any imbalance between these factors can lead to impaired bone healing, resulting in delayed union or non-union. To address this bone healing disorder, clinicians have adopted a technique known as "dynamization" which involves modifying the stiffness properties of the fixator. This technique facilitates the establishment of a favorable mechanical and biological environment by changing a rigid fixator to a more flexible one that promotes bone healing. However, the dynamization of fixators is selective for certain types of non-union and can result in complications or failure to heal if applied to inappropriate non-unions. This review aims to summarize the indications for dynamization, as well as introduce a novel dynamic locking plate and various techniques for dynamization of fixators (intramedullary nails, steel plates, external fixators) in femur and tibial fractures. Additionally, Factors associated with the effectiveness of dynamization are explored in response to the variation in dynamization success rates seen in clinical studies.

Use of network pharmacology and molecular docking to explore the mechanism of action of curcuma in the treatment of osteosarcoma.

Curcuma has been used as an adjuvant treatment for osteosarcoma (OS) due to its anticancer compounds. However, the underlying mechanism remains unclear. Therefore, this study aimed to explore the mechanism of action of curcuma in the treatment of OS using network pharmacology and molecular docking. In this study, anticancer compounds were obtained from relevant literature, and curcuma-related targets and OS treatment targets were obtained from public databases. Protein‒protein interaction networks were constructed to screen out the hub genes using the STRING database and Cytoscape software. Cluster analysis of the protein modules was then performed using the Cytoscape MCODE plugin. Furthermore, Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed for common targets among curcuma targets and OS-related targets using the DAVID database. Finally, molecular docking was performed, and the results were verified by Auto dock Tool and PyMOL software. Our research identified 11 potential active compounds, 141 potential therapeutic targets and 14 hub genes for curcuma. AKT1, TNF, STAT3, EGFR, and HSP90AA1 were the key targets closely related to the PI3K/Akt signaling pathways, HIF-1 signaling pathways, ErbB signaling pathways, and FOXO signaling pathways, which are involved in angiogenesis, cancer cell proliferation, metastasis, invasion, and chemotherapy resistance in the microenvironment of OS. Molecular docking suggested that the core compound had a strong affinity for key targets, with a binding energy of less than - 5 kJ/mol. The study showed that curcuma-mediated treatment of OS was a complex process involving multiple compounds, targets, and pathways. This study will enhance the understanding of how curcuma affects the proliferation and invasion of OS cells and reveal the potential molecular mechanism underlying the effect of curcuma on OS lung metastasis and chemotherapy resistance.

Effects of black soldier fly larvae as protein or fat sources on apparent nutrient digestibility, fecal microbiota, and metabolic profiles in beagle dogs.

Black soldier fly (Hermetia illucens) larvae (BSFL) act as a biological system converting organic waste into protein and fat with great potential application as pet food. To evaluate the feasibility of BSFL as a protein and fat source, 20 healthy beagle dogs were fed three dietary treatments for 65 days, including (1) a basal diet group (CON group), (2) a basal diet that replaced 20% chicken meal with defatted black soldier fly larvae protein group (DBP group), and (3) a basal diet that replaced 8% mixed oil with black soldier fly larvae fat group (BF group). This study demonstrated that the serum biochemical parameters among the three groups were within the normal range. No difference (p > 0.05) was observed in body weight, body condition score, or antioxidant capacity among the three groups. The mean IFN-γ level in the BF group was lower than that in the CON group, but there was no significant difference (p > 0.05). Compared with the CON group, the DBP group had decreasing (p < 0.05) apparent crude protein and organic matter digestibility. Furthermore, the DBP group had decreasing (p < 0.05) fecal propionate, butyrate, total short-chain fatty acids (SCFAs), isobutyrate, isovalerate, and total branched-chain fatty acids (BCFAs) and increased (p < 0.05) fecal pH. Nevertheless, there was no difference (p > 0.05) in SCFAs or BCFAs between the CON and BF groups. The fecal microbiota revealed that Lachnoclostridium, Clostridioides, Blautia, and Enterococcus were significantly enriched in the DBP group, and Terrisporobacter and Ralstonia were significantly enriched in the BF group. The fecal metabolome showed that the DBP group significantly influenced 18 metabolic pathways. Integrating biological and statistical correlation analysis on differential fecal microbiota and metabolites between the CON and DBP groups found that Lachnoclostridium, Clostridioides, and Enterococcus were positively associated with biotin. In addition, Lachnoclostridium, Clostridioides, Blautia, and Enterococcus were positively associated with niacinamide, phenylalanine acid, fumaric acid, and citrulline and negatively associated with cadavrine, putrescine, saccharopine, and butyrate. In all, 20% DBP restrained the apparent CP and OM digestibility, thereby affecting hindgut microbial metabolism. In contrast, 8% BF in the dog diet showed no adverse effects on body condition, apparent nutrient digestibility, fecal microbiota, or metabolic profiles. Our findings are conducive to opening a new avenue for the exploitation of DBP and BF as protein and fat resources in dog food.

Effect of Procyanidin on Canine Sperm Quality during Chilled Storage.

Procyanidin (PC) is a polyphenolic compound with antioxidant activity. The purpose of this study was to determine the influence of PC on canine sperm quality after 72 h of storage at 4 °C. The collected ejaculates were separated into four equal aliquots and treated with various concentrations of PC (0, 10, 30, and 50 µg/mL) in Tris-citric-fructose-egg yolk (TCFE) extender and stored at 4 °C for 72 h. The findings revealed that 30 µg/mL PC was the optimum concentration for significantly improving sperm motility (p < 0.05). Sperm samples treated with 30 µg/mL PC had substantially greater plasma membrane integrity, acrosome integrity, and mitochondrial membrane potential than the control group (p < 0.05). Furthermore, T-AOC and the expression levels of superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (GPx1) genes were significantly higher in sperm treated with 30 µg/mL PC than those in control (p < 0.05). In summary, this study discovered that adding PC to the TCFE extender enhanced sperm quality and that 30 µg/mL PC was the optimal concentration for canine sperm when stored at 4 °C.

The treatment of a Morel-Lavallée lesion of the thigh with incision and drainage along with tissue debridement and a surgically placed drain: A case report and literature review.

Background: A Morel-Lavallée lesion (MLL) is a rare closed degloving injury that usually occurs around the hips and is associated with pelvic fractures after high-energy trauma, which is commonly overshadowed by other severe post-traumatic manifestations. An isolated MLL, mostly caused by low-energy violence, is even rarer. Thus, the rates of misdiagnosis and missed diagnosis are often high. In this case report and literature review, we review the pathophysiology, clinical manifestations, imaging data, and treatment of this lesion to increase awareness of this rare disease. Case report: We report the case of an isolated MLL in the right thigh caused by trauma, which happened to be one of missed diagnosis both at the initial visit and at the return visit of the patient, with a significant sign of a mass on MRI. Given the size of the lesion, open debridement and irrigation were adopted to treat the lesion, and the patient recovered well post-operatively. Conclusion: Young surgeons should pay attention to the MLL with sufficient recognization to avoid missed diagnosis and misdiagnosis. Comprehensive physical examination and imaging data play important roles in the diagnosis of MLL. In the early stages of this injury, a detailed history review combined with physical examination and MRI, can reduce the rates of missed diagnosis and misdiagnosis. The choice of the therapeutic scheme depends on the size and severity of the lesion. For an isolated MLL, compared with conservative treatments, we suggest that incision and drainage, along with tissue debridement and a surgically placed drain, will reduce the rates of infection and recurrence.

Toxocara canis Infection Alters lncRNA and mRNA Expression Profiles of Dog Bone Marrow.

Bone marrow is the main hematopoietic organ that produces red blood cells, granulocytes, monocyte/macrophages, megakaryocytes, lymphocytes, and myeloid dendritic cells. Many of these cells play roles in the pathogenesis of Toxocara canis infection, and understanding how infection alters the dynamics of transcription regulation in bone marrow is therefore critical for deciphering the global changes in the dog transcriptional signatures during T. canis infection. In this study, long non-coding RNA (lncRNA) and messenger RNA (mRNA) expression profiles in the bone marrow of Beagle dogs infected with T. canis were determined at 12 h post-infection (hpi), 24 hpi, 96 hpi, and 36 days post-infection (dpi). RNA-sequencing and bioinformatics analysis identified 1,098, 984, 1,120, and 1,305 differentially expressed lncRNAs (DElncRNAs), and 196, 253, 223, and 328 differentially expressed mRNAs (DEmRNAs) at 12 h, 24 h, 96 h, and 36 days after infection, respectively. We also identified 29, 36, 38, and 68 DEmRNAs potentially cis-regulated by 44, 44, 51, and 80 DElncRNAs at 12 hpi, 24 hpi, 96 hpi, and 36 dpi, respectively. To validate the sequencing findings, qRT-PCR was performed on 10 randomly selected transcripts. Many altered genes were involved in the differentiation of bone marrow cells. GO of DElncRNAs and GO and KEGG pathway analyses of DEmRNAs revealed alterations in several signaling pathways, including pathways involved in energy metabolism, amino acid biosynthesis and metabolism, Wnt signaling pathway, Huntington's disease, HIF-1 signaling pathway, cGMP-PKG signaling pathway, dilated cardiomyopathy, and adrenergic signaling in cardiomyocytes. These findings revealed that bone marrow of T. canis-infected dogs exhibits distinct lncRNA and mRNA expression patterns compared to healthy control dogs. Our data provide novel insights into T. canis interaction with the definitive host and shed light on the significance of the non-coding portion of the dog genome in the pathogenesis of toxocariasis.

Psychometric analysis of a Chinese version of the Sleep Hygiene Index in nursing students in China: a cross-sectional study.

OBJECTIVE: The aim of the study had two stages. One was to examine the psychometric quality of the Sleep Hygiene Index (SHI) in Chinese version and its predicted function for the prevalence of insomnia. The other was to describe the prevalence of poor sleep hygiene habits and associated factors of sleep hygiene habits in Chinese nursing students. METHOD: According to Brislin translation model, the English version of SHI was translated into Chinese. And a pilot-survey was carried out to measure psychometric quality of the Chinese version of SHI with 260 nursing students by convenient sampling. Then a cross-sectional survey was conducted. 659 undergraduates were recruited by simple random sampling in a medical university in China. Data collection instruments consisted of a demographic questionnaire, the Sleep Hygiene Index (SHI), the Insomnia Severity Index (ISI) and the Brief Insomnia Questionnaire (BIQ). Data were analyzed by SPSS 24.0 and Amos 24.0 with P = 0.05 as the significant test value. RESULTS: The internal consistency reliability of SHI in Chinese version was more than 0.60 (α = 0.62, ω = 0.63). The concurrent validity presented significantly (r = 0.25, P < 0.001). Exploratory factor analysis found that a six component model explained 63.06% of total variance and confirmatory factor analysis showed good fitness (χ2/df=2.14, RMSEA = 0.04). ROC analysis showed that the cut-off value predicting for insomnia was 5.50 (52.90% sensitivity and 75.80% specificity). The area under the ROC curve was 0.66 (95% confidence interval = 0.61-0.71). 199 (30.20%) participant had poor sleep hygiene habits, especially in the aspects of staying too longer in bed (65.25%) and irregular sleep schedule. Multiple linear regression analysis showed health condition, academic difficulties and gender were more common associated factors of sleep hygiene. CONCLUSIONS: The Chinese version of the Sleep Hygiene Index demonstrates satisfactory psychometric qualities and has higher sensitivity and specificity to predict for insomnia. So SHI could be used in Chinese nursing students and detect high levels of insomnia. The status of sleep hygiene of nursing students in China should be concerned. Sleep hygiene education should be carried out in nursing students with different gender, health condition, and academic performance.

Global profiling of lncRNAs-miRNAs-mRNAs reveals differential expression of coding genes and non-coding RNAs in the lung of beagle dogs at different stages of Toxocara canis infection.

The roundworm Toxocara canis causes toxocariasis in dogs and larval migrans in humans. Better understanding of the lung response to T. canis infection could explain why T. canis must migrate to and undergoes part of its development inside the lung of the definitive host. In this study, we profiled the expression patterns of long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs in the lungs of Beagle dogs infected by T. canis, using high throughput RNA sequencing. At 24 h p.i., 1,012 lncRNAs, 393 mRNAs and 10 miRNAs were differentially expressed (DE). We also identified 883 DElncRNAs, 264 DEmRNAs and 20 DEmiRNAs at 96 h p.i., and 996 DElncRNAs, 342 DEmRNAs and eight DEmiRNAs at 36 days p.i., between infected and control dogs. Significant changes in the levels of expression of transcripts related to immune response and inflammation were associated with the antiparasitic response of the lung to T. canis. The remarkable increase in the expression of scgb1a1 at all time points after infection suggests the need for consistent moderation of the excessive inflammatory response. Also, upregulation of foxj1 at 24 h p.i., and downregulation of IL-1ß and IL-21 at 96 h p.i., suggest an attenuation of the humoral immunity of infected dogs. These results indicate that T. canis pathogenesis in the lung is mediated through contributions from both pro-inflammatory and anti-inflammatory mechanisms. Competing endogenous RNA (ceRNA) network analysis revealed significant interactions between DElncRNAs, DEmiRNAs and DEmRNAs, and improved our understanding of the ceRNA regulatory mechanisms in the context of T. canis infection. These data provide comprehensive understanding of the regulatory networks that govern the lung response to T. canis infection and reveal new mechanistic insights into the interaction between the host and parasite during the course of T. canis infection in the canine.

Proteomic alterations in the plasma of Beagle dogs induced by Toxocara canis infection.

Toxocara canis causes ocular larva migrans and visceral larva migrans in humans. Knowledge about the molecular mechanism of T. canis-hosts interaction is limited. The proteomic alterations in the plasma of Beagle dogs induced by T. canis infection were studied by the quantitative mass spectrometry-based data-independent acquisition (DIA). 418, 414 and 411 plasma proteins were identified at 24 h post-infection (hpi), 96 hpi and 36 days post-infection (dpi), including 6, 5 and 23 proteins with differential abundance, respectively. At 24 hpi, the altered proteins, retinoic acid receptor responder protein 2 (RARRES2), WD repeat-containing protein 1 (WDR1), moesin and filamin-A, may participate in pro-inflammatory reaction or promote larvae migration. At 96 hpi, the altered protein C and fibroleukin may maintain the stability of the coagulation system to protect the lung. At 36 dpi, the alterations of C-reactive protein (CRP), ficolin (FCN), complement factor H-related protein 5 (CFHR5) and other complements can affect the three traditional complement system, including the classic pathway, lectin pathway and alternative pathway. These proteins may play important roles in the interaction between T. canis and its definitive hosts. Further study on these altered proteins triggered by T. canis infection may discovery novel therapeutic or diagnostic targets for toxocariasis. SIGNIFICANCE OF THE STUDY: Toxocara canis is one of the globally distributed soil-transmitted helminths, which causes ocular larva migrans and visceral larva migrans in humans and a wide range of warm-blooded animals. T. canis adapts to different microenvironments by resisting and adjusting various biological processes of the hosts. Knowledge about the molecular mechanism of T. canis-hosts interaction is limited. Plasma proteins are good marker for monitoring the occurrence and development of diseases. The proteomic alterations in the plasma of Beagle dogs induced by T. canis infection were studied by the quantitative mass spectrometry-based data-independent acquisition (DIA) in this study. A total of 418, 414 and 411 plasma proteins were identified at 24 h post-infection (hpi), 96 hpi and 36 days post-infection, respectively. Ten protein with differential abundances were validated by using parallel reaction monitoring (PRM). Collectively, our deep proteomic analysis of plasma revealed that proteins alterations were affected by disease development, and proteomic analysis is an ideal method for quantifying changes in circulating factors on a global scale in response to pathophysiological perturbations such as T. canis infection.

Down-regulation of lncRNA UCA1 enhances radiosensitivity in prostate cancer by suppressing EIF4G1 expression via sponging miR-331-3p.

BACKGROUND: We aimed to explore the role of long noncoding RNA urothelial carcinoma-associated 1 (lncRNA UCA1) and its underlying mechanism in the radioresistance of prostate cancer (PCa). METHODS: QRT-PCR was conducted to measure the expression of UCA1, microRNA-331-3p (miR-331-3p) and eukaryotic translation initiation factor 4 gamma 1 (EIF4G1) in PCa tissues and cells. The relative protein level was determined by western blot assay. Cell proliferation and apoptosis were detected by MTT, colony formation assay, and flow cytometry, respectively. The target interaction between miR-331-3p and UCA1 or EIF4G1 was predicted through bioinformatics analysis, and verified by dual-luciferase reporter gene assay system. RESULTS: The high levels of UCA1 and EIF4G1 as well as the low level of miR-331-3p were observed in PCa tissues and cell lines. UCA1 and EIF4G1 expression were significantly upregulated by Gy radiation treatement. UCA1 or EIF4G1 knockdown repressed cell growth and enhanced cell apoptosis in 22RV1 and DU145 cells under radiation. Moreover, overexpression of EIF4G1 abolished UCA1 knockdown-induced effect on 6 Gy irradiated PCa cells. UCA1 sponged miR-331-3p to regulate EIF4G1 expression. CONCLUSIONS: LncRNA UCA1 deletion suppressed the radioresistance to PCa by suppressing EIF4G1 expression via miR-331-3p. UCA1 acted as a potential regulator of radioresistance of PCa, providing a promising therapeutic target for PCa.

Diagnostic and prognostic value of the cancer-testis antigen lactate dehydrogenase C4 in breast cancer.

BACKGROUND: Lactate dehydrogenase C4 (LDH-C4) as a cancer/testis antigen (CTA) is abnormally expressed in some malignant tumors. However, the expression and clinical significance of LDH-C4 in breast cancer (BC) has not been characterized. METHODS: We determined LDHC mRNA expression in serum and serum-derived exosomes of BC patients by quantitative RT-PCR. We also evaluated the protein expression of LDH-C4 in BC tissues using high-throughput tissue microarray analysis and immunohistochemistry. RESULTS: Our results showed high mRNA expression level of LDHC in serum and serum-derived exosomes of BC patients. The LDHC level in serum and exosomes could distinguish BC cases from healthy individuals based on their AUCs of 0.9587 and 0.9464, respectively. Besides, the LDHC level in exosomes of BC patients associated with tumor size, and positively correlated with HER2 and Ki-67 expressions (all with P < 0.05). Serum and exosomal level of LDHC negatively correlated with medical treatment and positively with the recurrence of BC. Survival analysis showed that LDH-C4 expression negatively correlated with BC prognosis. CONCLUSION: Serum and exosomal LDHC may be an effective indicator for the diagnosis, efficacy evaluation, and monitoring the recurrence of BC. LDH-C4 may act as a biomarker that predicts BC prognosis.

Effects of serum and follicular fluid on the in vitro maturation of canine oocytes.

The effects of gonadotropin, serum and follicular fluid on the in vitro maturation of canine oocytes were examined. Additionally, spindle size and spindle migration in MI-stage oocytes derived by in vivo or in vitro maturation were evaluated for the first time. Mature oocytes collected from beagle dog ovaries were divided into two experiments. In experiment I, oocytes were cultured in basic TCM 199 medium supplemented with different levels of P4, E2 and FSH. In experiment II, oocytes in the estrus or anestrus stage were cultured in basic medium supplemented with 30% or 40% canine serum plus 20% or 10% follicular fluid. Our results showed that in experiment I, more oocytes reached MI-MII (18.57%) after supplementation with 1 IU/ml FSH+ 5 IU/ml P4 + 5 IU/ml E2 than after supplementation with other levels of reagents. However, there were no significant differences among the groups (three different concentration groups and a control group) with respect to the proportions of oocytes that resumed meiosis, completed meiosis or degenerated. In experiment II, the number of oocytes from the estrus stage that reached MI-MII in TCM 199 medium supplemented with 40% canine serum and 10% follicular fluid (46.72%) was significantly higher (p < 0.01) than the number of oocytes from the anestrus stage that reached MI-MII in medium supplemented with 30% canine serum and 20% follicular fluid (21.84%). In addition, the degeneration rate was significantly lower (p < 0.05) in the 40% canine serum/10% follicular fluid group from follicular stage than in the other three groups. The average spindle length of the MI-stage oocytes that matured in vivo was significantly (p < 0.01) longer than that of the MI-stage oocytes that matured in vitro (21.75 vs. 14.39 µm). These results suggest that supplementation of the culture medium with 40% estrus serum and 10% follicular fluid had a positive influence on the in vitro maturation of canine oocytes and greatly affected spindle size in MI-stage oocytes.

Serum metabolomic alterations in Beagle dogs experimentally infected with Toxocara canis.

BACKGROUND: Toxocara canis, a globally distributed roundworm, can cause debilitating disease in dogs and humans; however, little is known about the metabolomic response of the hosts to T. canis infection. There is an increasing need to understand the metabolic mechanisms underlying the pathogenesis of T. canis infection in dogs. Here, we examined the metabolomic changes in Beagle dogs' serum following T. canis infection using LC-MS/MS. RESULTS: The metabolic profiles of Beagle dogs' serum were determined at 12 h, 24 h, 10 d and 36 d after oral infection with 300 infectious T. canis eggs by LC-MS/MS. We tested whether the T. canis-associated differentially abundant metabolites could distinguish the serum of infected dogs from controls, as measured by the area under the receiver operating characteristic (ROC) curve (AUC). The differentially expressed metabolites were further evaluated by principal components analysis and pathway enrichment analysis. A total of 5756 and 5299 ions were detected in ESI+ and ESI- mode, respectively. ROC curve analysis revealed nine and five metabolite markers, at 12 hpi and 24 hpi to 36 dpi, respectively, with potential diagnostic value for toxocariasis. The levels of taurocholate, estradiol, prostaglandins and leukotriene were significantly changed. Primary bile acid biosynthesis pathway, steroid hormone biosynthesis pathway and biosynthesis of unsaturated fatty acids pathway were significantly altered by T. canis infection. CONCLUSIONS: These findings show that T. canis infection can induce several changes in the dog serum metabolome and that the metabolic signature associated with T. canis infection in dogs has potential for toxocariasis diagnosis.

Diagnostic efficacy of long non-coding RNA MALAT-1 in human cancers: a meta-analysis study.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) is one kind of long non-coding RNAs (lncRNAs) that has been recognized as a hallmark of the onset and development of several carcinomas. This study seek to meta-analyze the overall diagnostic efficacy of elevated MALAT-1 expression profile for human cancers. Studies on the diagnostic performance of MALAT-1 in cancers were retrieved by searching the online databases. The combined effect sizes were summarized using a bivariate meta-analysis model. Impacts of publication bias on the pooled effect sizes were assessed using "Duval and Tweedie nonparametric trim and fill method". Sensitivity analysis and meta-regression test were applied to deeply trace the heterogeneity sources among eligible studies. A total of 14 studies with 1342 cancer cases were included. The combined effect sizes showed that MALAT-1 expression profiling conferred an estimated sensitivity of 0.69 (95% CI: 0.62-0.75) (I2 = 84.01%, P < 0.001), specificity of 0.85 (95% CI: 0.79-0.90) (I2 = 87.95%, P < 0.001) and AUC (area under curve) of 0.83 in distinguishing cancer patients from noncancerous contrasts. Moreover, stratified analysis depending on cancer type manifested that elevated MALAT-1 harbored a promising efficacy in the diagnosis of pulmonary tumors (AUC = 0.90), digestive system tumors (AUC = 0.84), gynecologic cancers (AUC = 0.84) and nasopharyngeal carcinoma (AUC = 0.84), particularly in confirming the subtype of squamous carcinoma (AUC = 0.91) and non-small cell lung carcinoma (AUC = 0.88) in lung cancer. Other analyses based on test matrix and ethnicity also presented robust results. Collectively, elevated MALAT-1 could be developed as an auxiliary molecular marker to aid in cancer diagnosis.

[Simultaneous determination of nine perfluorinated compound precursors in atmospheric precipitation by solid phase extraction and ultra performance liquid chromatography with tandem mass spectrometry].

A high-throughput detection method has been developed for the determination of nine perfluorinated compound precursors (PFCPs) in atmospheric precipitation by solid phase extraction-ultra performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (SPE-UPLC-ESI-MS/MS). The atmospheric precipitation samples were concentrated and purified with HLB solid phase extraction cartridges. The UPLC separation was performed on an HSS T3 column (100 mm×2.1 mm, 1.7 µm) utilizing a gradient elution program of methanol and water as the mobile phases at a flow rate of 0.2 mL/min. The MS/MS detection was performed under negative electrospray ionization (ESI-) in multiple reaction monitoring (MRM) mode. Good linearity was observed in the range of 0.05-5.00 µg/L, 0.50-50.0 µg/L or 5.00-500 µg/L with correlation coefficients from 0.9921 to 0.9995. The limits of detection (LODs) for the nine perfluorinated compound precursors were in the ranges of 0.05-7.9 ng/L. The recoveries ranged from 76.0% to 106% with the relative standard deviations between 0.72% and 13.7%. This method is characterized by high sensitivity and precision, extensive analytical range and quick analytical rate, and can be applied for the analysis of perfluorinated compound precursors in atmospheric precipitation.

PAX1 and SOX1 methylation as an initial screening method for cervical cancer: a meta-analysis of individual studies in Asians.

BACKGROUND: Epigenetic alterations of gene or DNA methylation have been highlighted as promising biomarkers for early cervical cancer screening. Herein, we evaluated the diagnostic performance of paired boxed gene 1 (PAX1) and sex determining region Y-box 1 (SOX1) methylation for cervical cancer detection. METHODS: Eligible studies were retrieved by searching the electronic databases. Study quality was assessed according to the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) checklist. The bivariate meta-analysis model was employed to plot the summary receiver operator characteristic (SROC) curve using Stata 12.0 software. RESULTS: The pooled sensitivity of PAX1 methylation was estimated to be 0.73 [95% confidence interval (CI): 0.70-0.75] in differentiating patients with HSIL (high-grade squamous intraepithelial lesion) or CIN3+ (cervical intraepithelial neoplasia type III/worse) or cervical cancer from normal individuals, corresponding to a specificity of 0.87 (95% CI: 0.85-0.89) and area under the curve (AUC) of 0.91. The SOX1 methylation test yielded an AUC of 0.82, under which, the pooled sensitivity was 0.71 (95% CI: 0.67-0.74) and specificity was 0.64 (95% CI: 0.61-0.67). Notably, the stratified analysis suggested that combing parallel testing of PAX1 methylation and human papillomavirus (HPV) DNA (AUC, sensitivity, and specificity of 0.89, 0.75, and 0.81, respectively) achieved higher accuracy than single HPV DNA testing (AUC, sensitivity, and specificity of 0.77, 0.81, and 0.70, respectively). CONCLUSIONS: PAX1 or SOX1 methylation has a prospect to be an auxiliary biomarker for cervical cancer screening, and parallel testing of PAX1 methylation and HPV DNA in cervical swabs confers an improved diagnostic accuracy than single HPV DNA testing.

Long noncoding RNAs as auxiliary biomarkers for gastric cancer screening: A pooled analysis of individual studies.

BACKGROUND: Long non-coding RNAs (lncRNAs) are highlighted as novel cancer biomarkers with great promise. Herein, we focused on summarizing the overall diagnostic performance of lncRNAs for gastric cancer (GC). METHODS: Publications fulfilling the search criteria were selected from the online databases. Study quality was assessed according to the Quality Assessment for Studies of Diagnostic Accuracy (QUADAS) checklist. The summary receiver operator characteristic (SROC) curve was plotted using a bivariate meta-analysis model. Statistical analysis was performed based on the platforms of STATA 12.0 and Meta-Disc 1.4 software. RESULTS: Fifteen studies with 1252 patients and 1283 matched controls were included. The pooled sensitivity and specificity for lncRNA expression profile in differentiating GC patients from cancer-free individuals were 0.68 (95%CI: 0.61-0.74) and 0.79 (95%CI: 0.72-0.84), respectively, corresponding to an area under curve (AUC) of 0.80. Moreover, the stratified analyses demonstrated that plasma-based lncRNA profiling harbored higher accuracy than that tissue-based assay (specificity: 0.80 versus 0.75; AUC: 0.84 versus 0.77). CONCLUSIONS: LncRNA profiling hallmarks a moderate diagnostic value in the management of GC and that lncRNA expression patterns may potentially be utilized as auxiliary biomarkers in confirming GC.