A GC×GC-MS method based on solid-state modulator for non-targeted metabolomics: Comparison with traditional GC-MS method.

The formidable challenge posed by the presence of extremely high amounts of compounds and large differences in concentrations in plasma significantly complicates non-targeted metabolomics analyses. In this study, a comprehensive two-dimensional gas chromatography-quadrupole mass spectrometry (GC×GC-qMS) method with a solid-state modulator (SSM) for non-targeted metabolomics in beagle plasma was first established based on a GC-MS method, and the qualitative and quantitative performance of the two platforms were compared. Identification of detected compounds was accomplished utilizing NIST database match scores, retention indices (RIs) and standards. Semi-quantification involved the calculation of peak area ratios to internal standards. Metabolite identification sheets were generated for plasma samples on both analytical platforms, featuring 22 representative metabolites chosen for validating qualitative accuracy, and for conducting comparisons of linearity, accuracy, precision, and sensitivity. The outcomes revealed a threefold increase in the number of identifiable metabolites on the GC×GC-MS platform, with lower limits of quantitation (LLOQs) reduced to 0.5-0.05 times those achieved on the GC-MS platform. Accuracy in quantification for both GC×GC-MS and GC-MS fell within the range of 85-115%, and the vast majority of intra- and inter-day precisions were within the range of 20%. These findings underscore that relative to the conventional GC-MS method, the GC×GC-MS method developed in this study, combined with SSM, exhibits enhanced qualitative capabilities, heightened sensitivity, and comparable accuracy and precision, rendering it more suitable for non-targeted metabolomics analyses.

A near-infrared fluorescent probe for viscosity: Differentiating cancer cells from normal cells and dual-modal imaging in tumor mice.

As a basic parameter of the intracellular microenvironment, viscosity is closely related to the development of cancer. Thus, it is necessary to utilize a sensitive tool to visualize the viscosity in tumor cells and mice, which is helpful for the diagnosis of cancer. Herein, a novel dual-modal probe (IX-V) that has a near-infrared fluorescence (NIRF) and photoacoustic (PA) response to viscosity is synthesized. In low viscosity media, the probe has no fluorescence. With the increase of viscosity, the fluorescence is produced in the near-infrared region due to the inhibition of the TICT process. At the same time, the probe shows different photoacoustic (PA) signals in different viscosity media. Most notably, the viscosity in tumor cells has been imaged successfully by the application of IX-V, and the probe can effectively distinguish cancer cells from normal cells co-cultured in one dish by the difference of fluorescence intensity. In addition, the probe has been used for dual-modal imaging (NIRF and PA) of viscosity in tumor mice, which provides a tool for exploring the relationship between viscosity and diseases. That is to say, IX-V can achieve complementary imaging effects and has great application prospects in the tumor diagnosis.

A near-infrared fluorescent probe for monitoring abnormal mitochondrial viscosity in cancer and fatty-liver mice model.

Viscosity is an important component of cell microenvironment, and abnormal mitochondrial viscosity is associated with many diseases such as tumor and fatty liver. Herein, a near-infrared fluorescence probe (QX-V) based on quinoline-xanthene dye for detecting viscosity is constructed. In high viscosity medium, the free rotation of single bond is inhibited and the fluorescence is released. The probe shows high sensitivity together with good selectivity. Notably, QX-V has a long excitation wavelength (710 nm) and emission wavelength (786 nm). At the same time, the probe is a positively charged molecule that can target mitochondria. QX-V can not only distinguish cancer cells from normal cells, but also make a distinction between normal cells and fatty hepatocytes. In addition, QX-V is used to image viscosity abnormality in tumor-bearing mice. The probe also has a good ability to image viscosity abnormality caused by liver injury in fatty-liver mice.

Global Trends and Future Prospects of Child Nutrition: A Bibliometric Analysis of Highly Cited Papers.

Child nutrition has always been a global concern. This study performed visual analysis of 1,398 child nutrition highly cited papers (HCPs) from 2009 to 2019. The purpose of the study was to evaluate and present the performances of authors, journals, countries, institutions, top cited papers; to explore the hot topics, prospects, and to propose the future research directions on child nutrition. We used bibliometric methods to conduct in-depth statistical analysis of HCPs on child nutrition, showing research progress, trends and hot spots. We included HCPs on child nutrition from the Science Citation Index-Expanded (SCI-E) database February 7, 2020. Two tools, CiteSpace and VOSviewer, were used to conduct the bibliometric analyses. The results showed that, since 2011, the number of HCPs on child nutrition has increased rapidly. The top three contributors in this field were the USA, the UK and Canada. However, the contribution of developing countries was very limited. Intestinal microflora, food allergy, overweight and obesity were the three major research hotspots in this field. Results of this study provide valuable references for ongoing child nutrition related research, which may be interesting and noteworthy to the researchers involved.

Nationwide assessment of persistent halogenated compounds (PHCs) in farmed golden pompano of China.

Persistent halogenated compounds (PHCs) contamination has become a major concern over the world. Here we investigated occurrence, spatial distributions, congener profiles, as well as health risks of PHCs in farmed golden pompano in China using gas chromatography/mass spectrometry (GC/MS). The concentrations of PCBs, PBDEs and OCPs were in the range of 0.78-4.79 ng/g wet weight (ww), not detected (nd)-1.14 ng/g ww and 1.1-38.8 ng/g ww, respectively. Furthermore, ρ,ρ'-DDT, ο,ρ'-DDT and PCB 101 were the dominant PHC contaminants. The estimated daily intakes of PHCs through consumption of golden pompano were up to 12.86 and 131.34 ng/kg body weight/day based on the mean and 95th concentrations determined in golden pompano, respectively. Risk-based analysis indicates that target PHCs in golden pompano would not pose risks to human. Our study presents the first report of a nationwide survey of PHCs contamination in farmed golden pompano in China.

[Correlation between high-risk TP53 mutation and extracapsular spread in oral squamous cell carcinoma].

PURPOSE: To investigate the correlation between high-risk tumor protein p53 (TP53) mutation and extracapsular spread(ECS) in oral squamous cell carcinoma (OSCC). METHODS: The data of 88 OSCC patients admitted to Gansu Provincial People's Hospital from January 2013 to January 2016 were retrospectively analyzed. The patients were divided into non-lymph node metastasis group (A1), lymph node metastasis without ECS group (A2), lymph node metastasis with ECS group(A3) according to the presence or absence of lymph node metastasis. The deletion of exon 5-8 of TP53 gene in primary and metastatic lesions was detected. The correlation of ECS with disease-free survival(DFS), overall survival(OS) rate and TP53 mutation were determined, and single factor analysis of ECS was performed. The data were analyzed by SPSS 20.0 software package. RESULTS: TP53 (5) homozygote deletion was found in all three groups. TP53 (6) homozygous deletion was found in group A3 (10/30). No homozygous deletion of TP53(7) was found in three groups.Homozygous deletion of TP53 (8) was found in group A2(7/42), group A3(10/30) and group A3(10/60).The 1-year and 3-year DFS rates were 75.00% and 70.83% in group A1, 70.59% and 58.82% in group A2, 46.67% and 40.00% in group A3, with significant difference(P<0.05). The 1-year and 3-year OS rates were 83.33% and 75.00% in group A1, 73.53% and 50.00% in group A2, 46.67% and 40.00% in group A3 , with significant difference(P<0.05). In group A3, there were 4 cases (13.33%) of low-risk TP53 mutation (LR), 12 cases (40.00%) of high-risk TP53 mutation (HR), 4 cases(13.33%) of wild-type TP53 mutation (Wt), and 10 cases(33.33%) of other mutations. HR mutation and other mutations occurred more frequently than other types(P<0.05). Smoking, primary lesion size, wild type/low risk and high risk/others were correlated with ECS(P<0.05). CONCLUSIONS: ECS is an important marker of DFS and OS in OSCC patients, and high-risk mutations were common in ECS, indicating a certain correlation between high-risk mutations of TP53 and ECS in OSCC.

Whole genome characterization of a novel porcine reproductive and respiratory syndrome virus 1 isolate: Genetic evidence for recombination between Amervac vaccine and circulating strains in mainland China.

Genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV 1) have been continuously isolated in China in recent years. Complete genome sequences of these isolates are important to investigate the prevalence and evolution of Chinese PRRSV 1. Herein, we describe the isolation of a novel PRRSV 1 isolate, denominated HLJB1, in the Heilongjiang province of China. Complete genome sequencing of HLJB1 showed that it shares 90.66% and 58.21% nucleotide identities with PRRSV 1 and 2 prototypic strains Lelystad virus and ATCC VR-2332, respectively. HLJB1 has a unique 5-amino-acid insertion in nsp2, which has never been described in other PRRSV 1 isolates. Whole genome-based phylogenetic analysis revealed that all Chinese PRRSV 1 isolates are clustered in pan-European subtype 1 and can be divided into four subgroups. HLJB1 resides in the subgroup of BJEU06-1-like isolates but is also closely related to the Amervac-like isolates. Additionally, recombination analyses suggested that HLJB1 is a recombinant from the Amervac vaccine and the BJEU06-1 isolate. To our best knowledge, our results provide the first genetic evidence for recombination between Amervac vaccine and circulating strains. These findings are also beneficial for studying the origin and evolution of PRRSV 1 in China.

Anti-Recombinant Gametocyte 56 Protein IgY Protected Chickens from Homologous Coccidian Infection.

Coccidiosis is caused by intra-cellular infection of Eimeria spp., which goes through a complex life cycle in the intestinal mucosa of infected hosts. Specific immunoglobulins (IgY) could be produced in egg yolk by immunizing hens with specific antigens. In the present study, we cloned the E. maxima gam56 gene, expressed the GST-GAM56 fusion protein and raised IgY to GST-GAM56 in hens. The anti-GST-GAM56 IgY antibody was isolated and used to treat chickens infected with E. maxima oocysts. Intramuscular injection of the antibodies provided minimal protection against parasite infection. However, oral dosing of the IgY 3 or 5 d after oocyst inoculation significantly improved body weight gain, reduced oocyst output and intestinal lesion score were reduced at 3 or 5 d after oocyst challenging, compared to the untreated control group. Our findings suggest that the IgY to gam56 could be an effective prophylactic or therapeutic agent against E. maxima infection in chickens and should have a practical application value.

Multi-epitope recombinant vaccine induces immunoprotection against mixed infection of Eimeria spp.

Immunity to Eimeria is species-specific, and chickens with immunity to one species of Eimeria remain susceptible to other Eimeria species. This presents a major challenge in the development of effective vaccines against multiple Eimeria species. In this study, we cloned the antigenic epitope of a tachyzoite surface protein gene of Eimeria tenella, a tachyzoite surface protein gene of Eimeria acervulina and the gametocyte protein gene of Eimeria maxima, and constructed prokaryotic and eukaryotic plasmids carrying the multi-epitope antigenic gene. Immunization of chickens with the multivalent DNA and protein conferred partial protection against infection by the three Eimeria species, as shown by increased CD4+ T lymphocytes in the intestinal mucosa, decreased oocyst excretion and intestinal lesions, and increased body weight gain compared with non-immunized controls. The DNA prime-protein boost immunization schedule induced greater cellular immunity and protection from Eimeria infection than immunization with DNA or protein alone. Our findings demonstrated that DNA prime-protein boost immunization with a multivalent vaccine could stimulate protective immunity against challenge infection of multiple Eimeria species. This work provides a promising step towards DNA-protein vaccination against multiple species of pathogens.