Isolation, identification, molecular and pathogenicity characteristics of an infectious laryngotracheitis virus from Hubei province, China.

Multiple outbreaks of avian infectious laryngotracheitis (ILT) in chickens, both domestically and internationally, have been directly correlate to widespread vaccine use in affected countries and regions. Phylogenetic and recombination event analyses have demonstrated that avian infectious laryngotracheitis virus (ILTV) field strains are progressively evolving toward the chicken embryo-origin (CEO) vaccine strain. Even with standardized biosecurity measures and effective prevention and control strategies implemented on large-scale farms, continuous ILT outbreaks result in significant economic losses to the poultry industry worldwide. These outbreaks undoubtedly hinder efforts to control and eradicate ILTV in the future. In this study, an ILTV isolate was successfully obtained by laboratory PCR detection and virus isolation from chickens that exhibited dyspnea and depression on a broiler farm in Hubei Province, China. The isolated strain exhibited robust propagation on chorioallantoic membranes of embryonated eggs, but failed to establish effective infection in chicken hepatocellular carcinoma (LMH) cells. Phylogenetic analysis revealed a unique T441P point mutation in the gJ protein of the isolate. Animal experiments confirmed the virulence of this strain, as it induced mortality in 6-wk-old chickens. This study expands current understanding of the epidemiology, genetic variations, and pathogenicity of ILTV isolates circulating domestically, contributing to the elucidate of ILTV molecular basis of pathogenicity and development of vaccine.

Circular RNA hsa_circ_0008003 facilitates tumorigenesis and development of non-small cell lung carcinoma via modulating miR-488/ZNF281 axis.

As one of the most aggressive malignancies, non-small cell lung carcinoma (NSCLC) has high risks of death. It has been demonstrated that circRNAs accelerate NSCLC progression, but the underlying molecular mechanisms of circRNAs in NSCLC were still obscure. In the first place, the circRNA microarray of NSCLC was investigated in this study, and hsa_circ_0008003 (circ-0008003) was chosen as the research object. Then, it was unveiled that the expression of circ-0008003 examined via qRT-PCR was elevated in tumour tissues relative to the non-tumour tissues, which was associated with TNM stage and lymphatic metastasis in NSCLC. Additionally, the prognosis of NSCLC patients with high circ-0008003 level was poor. Besides, circ-0008003 silencing dampened the invasion and proliferation of NSCLC cells. Next, according to the mechanistic studies, circ-0008003 functioned as a ceRNA of ZNF281 in NSCLC by acting as the endogenous sponge for miR-488, which was proved to be a tumour suppressor in NSCLC. Additionally, ZNF281 overexpression and miR-488 suppression recovered the influences of repressed circ-0008003 on NSCLC cellular processes. It was validated in this research that circ-0008003 triggered tumour formation in NSCLC, which was adjusted via miR-488/ZNF281 axis, casting a novel light on the resultful target for treating NSCLC and predicting the prognosis.

Circulating microRNA signatures serve as potential diagnostic biomarkers for Helicobacter pylori infection.

Helicobacter pylor (H pylori), a Gram-negative, microaerobic human pathogen, has been found to be involved in many gastroduodenal diseases. Accurate diagnosis of H pylori infection is a vital part of the effective management of gastroduodenal diseases. Circulating microRNAs (miRNAs) have shown the potential to be used as noninvasive biomarkers for the diagnosis of infectious diseases. The aim of this study was to explore plasma miRNAs as noninvasive biomarkers for H pylori infection. We performed a plasma miRNA expression profile using Illumina high-throughput sequencing and validated the levels of differentially expressed miRNAs in the plasma of 63 H pylori-infected patients and 41 healthy volunteers by quantitative real-time polymerase chain reaction (qRT-PCR). The sequencing results showed that 37 miRNAs were upregulated in the H pylori-infected patients compared with that in the healthy volunteers, while six miRNAs were downregulated. qRT-PCR and receiver operator characteristic analysis suggested that the expression of miR-28-3p, miR-143-3p, miR-151a-3p, and miR-148a-3p were closely associated with H pylori infection. Therefore, the four plasma miRNA panels mentioned above could serve as promising noninvasive biomarkers of H pylori infection.

Simple and sensitive LC-MS/MS method for simultaneous determination of crizotinib and its major oxidative metabolite in human plasma: Application to a clinical pharmacokinetic study.

In this study, a fast, simple and sensitive liquid chromatography-mass spectrometry method was developed for simultaneous determination of crizotinib and its major oxidative metabolite crizotinib-lactam in human plasma. The plasma samples were deproteinated by using acetonitrile containing 0.1% formic acid as precipitant whereas the chromatographic separation was obtained on a C18 column with 0.1% formic acid aqueous and acetonitrile/methanol (v:v, 1:1) as mobile phase. The mass detector was operated in positive selected reaction monitoring mode. Precursor-to-product transitions were optimized to be m/z 450.1/260.1, m/z 464.1/98.1, and m/z 326.1/291.1 for crizotinib, crizotinib-lactam and midazolam (internal standard), respectively. The established method was validated in accordance with guidance issued by Food and Drug Administration. The assay showed good linearity over the concentration ranges of 0.1-1000 ng/mL for crizotinib and 0.1-400 ng/mL for crizotinib-lactam, with correlation coefficients more than 0.999 (r > 0.999). The extraction recovery was more than 87.12%. No significant matrix effect and carryover were observed. The precision (RSD, %) was less than 8.27%, whereas accuracy (RE, %) was within the range of -4.56 to 7.08%. The validated method has been successfully applied to the clinical pharmacokinetic study of crizotinib and crizotinib-lactam in human plasma after oral administration of crizotinib at a single dose of 250 mg. The results revealed that crizotinib was rapidly metabolized into its metabolite crizotinib-lactam and the in vivo exposure of crizotinib-lactam was 38.50% of that of crizotinib.

The two-component system NisK/NisR contributes to the virulence of Streptococcus suis serotype 2.

Two-component signal-transduction systems (TCSTSs) may regulate some virulence factors in response to external stimuli, and thus allowing Streptococcus suis serotype 2 to interact with the host, promote survival, and cause disease. Here, a mutant of the NisKR TCSTS had attenuated virulence in vitro, as exemplified by lowered hemolytic activity, reduced adherence to epithelial cells, increased elimination by macrophages, and decreased resistance to killing by neutrophils. Results also showed that this system is important for the ability of S. suis serotype 2 to survive and proliferate in an in vivo mouse model. Thus, the NisKR system plays a significant role in pathogenesis, both in colonization and invasive disease.