Comparative transcriptomics in the hypothalamic-pituitary-gonad axis of mammals and poultry.

The hypothalamic-pituitary-gonad (HPG) axis is vital for reproductive activities in vertebrates. The large-scale comparative analyses of gene expression in the HPG axis across vertebrates have not been carried out yet. Here we collected 175 high-quality RNA-seq samples of hypothalamus, pituitary, ovary and testis from eight species (four mammals and four poultry) to compare transcriptome in the HPG axis, and to detect key pathways and related genes associated with reproduction. We demonstrated the distinguished difference in gene expression of the HPG axis between mammalian and avian species by a series of bioinformatics analysis, including gene differential expression, the phylogeny analysis of gene expression, and their functional annotations. We revealed two pathways, i.e., neuroactive ligand-receptor interaction and calcium signaling pathway, which play important roles in animal reproduction. In these two pathways, we detected 17 differentially expressed genes shared in 4 tissues, while 13, 27, and 27 were specifically differentially expressed genes in hypothalamus, pituitary and ovary, respectively. Our study on the comparative transcriptomics in the HPG axis across species will provide novel knowledge for exploring the molecular mechanism underlying reproductive traits in animals.

DNA Demethylation of Myogenic Genes May Contribute to Embryonic Leg Muscle Development Differences between Wuzong and Shitou Geese.

Skeletal muscle development from embryonic stages to hatching is critical for poultry muscle growth, during which DNA methylation plays a vital role. However, it is not yet clear how DNA methylation affects early embryonic muscle development between goose breeds of different body size. In this study, whole genome bisulfite sequencing (WGBS) was conducted on leg muscle tissue from Wuzong (WZE) and Shitou (STE) geese on embryonic day 15 (E15), E23, and post-hatch day 1. It was found that at E23, the embryonic leg muscle development of STE was more intense than that of WZE. A negative correlation was found between gene expression and DNA methylation around transcription start sites (TSSs), while a positive correlation was observed in the gene body near TTSs. It was also possible that earlier demethylation of myogenic genes around TSSs contributes to their earlier expression in WZE. Using pyrosequencing to analyze DNA methylation patterns of promoter regions, we also found that earlier demethylation of the MyoD1 promoter in WZE contributed to its earlier expression. This study reveals that DNA demethylation of myogenic genes may contribute to embryonic leg muscle development differences between Wuzong and Shitou geese.

Novel mutations in the Myo5a gene cause a dilute coat color phenotype in mice.

C57BL/6 laboratory mice usually show black coat color. We observed a dilute (gray) coat color phenotype in progenies of two C57BL/6 mice. This phenotype is inherited in an autosomal recessive mode. To uncover the molecular mechanism underlying this naturally occurring phenotypic variation, we performed whole-genome sequencing (25×) on 10 offspring of the two founder mice. The whole-genome DNA sequencing and additional RNA-Seq data reveal that Myo5a is the gene responsible for the coat color dilution in C57BL/6 mice, and novel mutations in the Myo5a gene are likely causal. We further performed reverse transcription-quantitative PCR, and showed increased expression of truncated Myo5a transcripts encoding dysfunctional proteins and decreased expression of Myo5a full-length transcripts encoding functional proteins in mutant individuals. The decrease in full-length messenger RNA abundance was accompanied by reduced Myo5a protein level and deficient melanosome transport, a potential mechanistic link between the Myo5a mutations and the dilute color phenotype. This study not only advances our understanding of the molecular mechanisms of pigmentation in mice, but also provides a typical case of deciphering the molecular basis of phenotypic variation in mice by genomic analyses and subsequent functional work.

Quality inspection and result analysis of the spirometer calibration cylinder.

BACKGROUND: To understand the accuracy of volume calibration syringes used in China and compare the difference between new and old volume calibration syringes, technical testing was performed on volume calibration syringes in clinical lung function instruments. MATERIALS AND METHODS: A standard validator device (Model 1180, Hans Rudolph, USA) was used to perform leak testing and volume accuracy testing for calibration syringes. Sixteen volume calibration syringes from 8 brands (CareFusion in Germany, Vyaire in Germany, Yaeger in Germany, Vitalograph in the United Kingdom, MGC Diagnostics in the United States, U-Breath in Zhejiang, China, Wendi in Ningbo, Zhejiang, and Boya in Ningbo, China) were tested. RESULTS: A total of 75% (12/16) of the volume calibration syringes passed the pressure decay leak test, 69% (11/16) of the volume calibration syringes passed the volume accuracy and repeatability test, and 56% (9/16) passed both tests; there was no significant difference in the total passing of the new and old volume calibration syringe quality tests (P > 0.05). CONCLUSIONS: A standard validator device should be used for both leakage tests and volume accuracy and repeatability tests to ensure the reliability of volume calibration syringes. It is suggested that the quality verification of volume calibration syringes should be regularly conducted to ensure the accuracy of the pulmonary function tests.

Technical performance analysis of different types of spirometers.

BACKGROUND: The spirometer is an important element in lung function examinations, and its accuracy is directly related to the accuracy of the results of these examinations and to the diagnosis and treatment of diseases. Our aim was to conduct a performance analysis of the detection techniques of differential pressure and ultrasonic portable spirometers commonly used in China. METHODS: A standard flow/volume simulator was used to analyze the performance (accuracy, repeatability, linearity, impedance, and so on) of portable spirometers, 4 imported and 6 domestic, based on 13 curves generated by different air sources in the ISO 26782:2009 standard. A Bland-Altman diagram was used to evaluate the consistency between the values measured by the spirometers and the simulator. RESULTS: The pass rates for accuracy, repeatability, linearity, and impedance for the 10 different portable spirometers were 50%, 100%, 70%, and 70%, respectively. Only 30% (3/10) of the spirometers-2 domestic and 1 imported-met all standards of quality and performance evaluation, while the rest were partially up to standard. In the consistency evaluation, only 3 spirometers were within both the consistency standard range and the acceptability range. CONCLUSION: The quality and performance of different types of portable spirometers commonly used in the clinic differ. The use of a standard flow/volume simulator is helpful for the standard evaluation of the technical performance of spirometers.

Distribution Pattern of Organophosphate Esters in Particle-Size Fractions of Urban Topsoils Under Different Land-Use Types and Its Relationship to Organic Carbon Content.

In this study, the distribution pattern of organophosphate esters (OPEs) in particle-size fractions of urban topsoils under different land-use types and its relationship to organic carbon content was investigated. Total OPEs concentrations in different particle-size fractions ranged from 17.07 to 221.77 ng/g. The distribution pattern of total OPEs concentrations and individual OPE concentration in different particle-size fractions were irregular and varied with the land-use type. The mass of OPEs is concentrated in small particles, large particles, or evenly distributed in each particle. This distribution pattern mainly depends on the mass distribution of each fraction to the soil. Tri-iso-butyl phosphate, tributyl phosphate, and triphenylphosphine oxide have a relatively higher concentration in most samples, and the concentration of tripropyl phosphate was the lowest in all samples. The correlations between total OPEs concentrations versus total organic carbon (TOC), black carbon (BC), and other carbon (OC) is weak. Their linear regression correlation coefficients were 0.0495, 0.0823, and 0.0097, respectively. The correlation between individual OPE concentrations versus TOC, BC, and OC also are weak. Except for triethyl phosphate, tris-(2-chloroethyl) phosphate, and tris-(1-chloro-2-propyl) phosphate, the linear regression correlation coefficients of other OPEs are all less than 0.1.

Trans-cinnamaldehyde shows anti-depression effect in the forced swimming test and possible involvement of the endocannabinoid system.

Depression is a mental disease that significantly reduces the quality of patients' life. Around 322 million people of all ages carry the heavy burden of depression on a worldwide scale, with a life-time prevalence of 20% according to the WHO. Trans-cinnamaldehyde (TCA) is an excellent COX-2 inhibitor in central nervous system which is a main constituent of GUIZHI as a member of traditional Chinese herb. Furthermore, previous studies demonstrated that TCA suppressed depression-like behavior in chronic unexpected mild stress, plus maze test and open field test. However, the molecular mechanism of TCA anti-depression effect is not clear. We examined the immobility of TCA pretreated male BALB/c mice in the forced swimming test (FST). Results show that TCA (50 mg/kg, po) revealed a significant effect on reduced immobility in the FST, compared with SAL group which indicated that TCA suppressed depression-like behavior. Moreover, TCA elevated the level of 5-HT and decreased the ratio of Glu/GABA in mice hippocampus. Compared with SAL + FST group, TCA + FST group significantly decreased COX-2, TRPV1 and CB1 protein level in mice hippocampus (p < 0.05, p < 0.05, p < 0.01). These findings suggest that TCA treatment exerted anti-depressive effect and was able to regulate neurotransmitters in the FST. This effect may have positive influence on the endocannabinoid (eCB) system.

Identification of copy number variations using high density whole-genome SNP markers in Chinese Dongxiang spotted pigs.

OBJECTIVE: Copy number variations (CNVs) are a major source of genetic diversity complementary to single nucleotide polymorphism (SNP) in animals. The aim of the study was to performed a comprehensive genomic analysis of CNVs based on high density whole-genome SNP markers in Chinese Dongxiang spotted pigs. METHODS: We used customized Affymetrix Axiom Pig1.4M array plates containing 1.4 million SNPs and the PennCNV algorithm to identify porcine CNVs on autosomes in Chinese Dongxiang spotted pigs. Then, the next generation sequence data was used to confirm the detected CNVs. Next, functional analysis was performed for gene contents in copy number variation regions (CNVRs). In addition, we compared the identified CNVRs with those reported ones and quantitative trait loci (QTL) in the pig QTL database. RESULTS: We identified 871 putative CNVs belonging to 2,221 CNVRs on 17 autosomes. We further discarded CNVRs that were detected only in one individual, leaving us 166 CNVRs in total. The 166 CNVRs ranged from 2.89 kb to 617.53 kb with a mean value of 93.65 kb and a genome coverage of 15.55 Mb, corresponding to 0.58% of the pig genome. A total of 119 (71.69%) of the identified CNVRs were confirmed by next generation sequence data. Moreover, functional annotation showed that these CNVRs are involved in a variety of molecular functions. More than half (56.63%) of the CNVRs (n = 94) have been reported in previous studies, while 72 CNVRs are reported for the first time. In addition, 162 (97.59%) CNVRs were found to overlap with 2,765 previously reported QTLs affecting 378 phenotypic traits. CONCLUSION: The findings improve the catalog of pig CNVs and provide insights and novel molecular markers for further genetic analyses of Chinese indigenous pigs.

Development of a field sampling method based on magnetic nanoparticles for the enrichment of pesticides in aqueous samples.

A field sampling method based on magnetic core-shell silica nanoparticles was developed for field sampling and the enrichment of low concentrations of pesticides in aqueous samples. The magnetic nanoparticles could be easily extracted from water samples by a custom-made magnetic nanoparticle collector. The recovery of 15 mg of magnetic particles from a 500 mL water sample was 90.8%. Mixtures of seven pesticides spiked into pure water and pond water were used as marker samples to evaluate the field sampling method. The average recoveries at three levels of spiking were in the range 60.0-104.7% with relative standard deviations <7.1%. The proposed method has good linearity with a correlation coefficient >0.9990 in the concentration range 0.5-15 µg L(-1). The results of the analysis of a sample of poisoned pond water indicate that this method is fast, convenient and efficient for the field sampling and enrichment of pesticides in aqueous samples.

Circular RNA-mediated miRNA sponge & RNA binding protein in biological modulation of breast cancer.

Circular RNAs (circRNAs) and small non-coding RNAs of the head-to-junction circle in the construct play critical roles in gene regulation and are significantly associated with breast cancer (BC). Numerous circRNAs are potential cancer biomarkers that may be used for diagnosis and prognosis. Widespread expression of circRNAs is regarded as a feature of gene expression in highly diverged eukaryotes. Recent studies show that circRNAs have two main biological modulation models: sponging and RNA-binding. This review explained the biogenesis of circRNAs and assessed emerging findings on their sponge function and role as RNA-binding proteins (RBPs) to better understand how their interaction alters cellular function in BC. We focused on how sponges significantly affect the phenotype and progression of BC. We described how circRNAs exercise the translation functions in ribosomes. Furthermore, we reviewed recent studies on RBPs, and post-protein modifications influencing BC and provided a perspective on future research directions for treating BC.

Curcumin for Treating Breast Cancer: A Review of Molecular Mechanisms, Combinations with Anticancer Drugs, and Nanosystems.

Breast cancer (BC) has become the fifth most prevalent cause of cancer-related morbidity, attracting significant attention from researchers due to its heightened malignancy and drug resistance. Conventional chemotherapy approaches have proven inadequate in addressing all BC subtypes, highlighting the urgent need for novel therapeutic approaches or drugs. Curcumin (CUR), a phytochemical derived from Curcuma longa (turmeric), has shown substantial potential in inhibiting BC cell migration, metastasis, and proliferation. However, the use of CUR in this context comes with challenges due to its dynamic and easily degradable nature, poor aqueous solubility, low bioavailability, rapid metabolism, and swift systemic elimination, collectively limiting its clinical applications. As such, we provide an overview of the properties, synthesis, and characterization of the hybridization of CUR and its analogue with chemo-drug building blocks. We reviewed research from the last five years on CUR's biogenesis with respect to the regulation of BC, revealing that CUR participates in arresting BC cells in the cell cycle and significantly induces apoptosis in BC cells. Information on the chemotherapeutic and antitumor mechanisms of CUR in BC, including regulation of the cell cycle, increased cell apoptosis, and inhibition of multidrug resistance (MDR), was compiled. Additionally, we provide an overview of CUR loaded into nanomaterials that are cotreated with other chemotherapeutic drugs, such as paclitaxel, thymoquinone, and tamoxifen. In this review, we discuss different types of nanoparticles that can be used for CUR delivery, such as polymeric nanoparticles, carbon nanotubes, and liposomes. By comparing the size, entrapment efficiency, drug-loading capacity, release time, biocompatibility, pharmaceutical scale, and reproducibility of various nanomaterials, we aimed to determine which formulations are better suited for loading CUR or its analogue. Ultimately, this review is expected to offer inspiring ideas, promising strategies, and potential pathways for developing advanced anti-BC strategy nanosystems in clinical practice.

Identification of Differentially Expressed Genes and microRNAs in the Gray and White Feather Follicles of Shitou Geese.

The Shitou goose, a highly recognized indigenous breed with gray plumage originating from Chaozhou Raoping in Guangdong Province, China, is renowned for being the largest goose species in the country. Notably, during the pure breeding process of Shitou geese, approximately 2% of the offspring in each generation unexpectedly exhibited white plumage. To better understand the mechanisms underlying white plumage color formation in Shitou geese, we conducted a comparative transcriptome analysis between white and gray feather follicles, aiming to identify key genes and microRNAs that potentially regulate white plumage coloration in this unique goose breed. Our results revealed a number of pigmentation genes, encompassing TYR, TYRP1, EDNRB2, MLANA, SOX10, SLC45A2, GPR143, TRPM1, OCA2, ASIP, KIT, and SLC24A5, which were significantly down-regulated in the white feather follicles of Shitou geese. Among these genes, EDNRB2 and KIT emerged as the most promising candidate genes for white plumage coloration in Shitou geese. Additionally, our analysis also uncovered 46 differentially expressed miRNAs. Of these, miR-144-y may play crucial roles in the regulation of feather pigmentation. Furthermore, the expression of novel-m0086-5p, miR-489-y, miR-223-x, miR-7565-z, and miR-3535-z exhibits a significant negative correlation with the expression of pigmentation genes including TYRP1, EDNRB2, MLANA, SOX10, TRPM1, and KIT, suggesting these miRNAs may indirectly regulate the expression of these genes, thereby influencing feather color. Our findings provide valuable insights into the genetic mechanisms underlying white plumage coloration in Shitou geese and contribute to the broader understanding of avian genetics and coloration research.

Exploration of quality criteria for the detection of peak inspiratory flow under different resistance conditions.

BACKGROUND: At present, robust quality criteria and methods for the assessment of Peak inspiratory flow meter performance are lacking. OBJECTIVE: A standard flow-volume simulator for quality control analyses of an inhalation assessment device was utilized with different simulated resistance levels in order to propose a quality testing method and associated standard for this device type. METHODS: A standard flow-volume simulator was utilized to assess the performance of an In-Check DIAL® (Device I) and an intelligent inhalation assessment device (Device P) at a fixed volume and flow rate. Indices used to evaluate these two instruments included repeatability, accuracy, linearity, and impedance. RESULTS: Both devices exhibited good repeatability (<± 3 L/min). The difference between test results and standard simulator values for Device P was less than ± 5 L/min at resistance level R1 but higher than ± 5 L/min at resistance levels R2-5, while Device I were greater than 5 L/min at all resistance levels. The relative error for Device P was <± 10% at resistance levels R1, R2, and R4, but > 10% at resistance levels R3 and R5. The relative error values for Device I at all five resistance levels were > 10%. Device P passed the linearity test at the R2 resistance level, while Device I partially passed the linearity test at all five resistance levels. CONCLUSION: Standard monitoring methods and standards provide a valuable approach to the more reliable clinical assessment and application of these instruments.

Assessing the effectiveness of inhalation therapy in patients with chronic airway diseases: A new digital measurement.

BACKGROUND: This study aimed to evaluate the effectiveness of inhalation therapy in patients with chronic airway diseases via the use of a new multiparametric inhalation assessment device. METHODS: A multiparametric inhalation evaluation device (PF810, UBREATH, Zhejiang, China) that could simulate common inhalation devices with 6 different levels (0-V) of resistance was used in this study. The device was considered suitable if the three parameters of peak inspiratory flow rate (PIFR), effective inspiratory time (EIT), and breath-hold time (BHT) after inspiration met the minimum requirements. RESULTS: A total of 4,559 tests were performed. The qualification rates of 0-V resistance gear from low to high were 3.38 % (I), 8.42 % (0), 15.31 % (II), 16.71 % (III), 20.27 % (IV), and 46.91 % (V). The COPD patients in the 3 experimental groups had the lowest percentages of isolates classified as resistant 0, III, and V, which were 5.65 %, 11.93 %, and 40.43 %, respectively. The lowest percentage was 39.67 % (V) for insufficient EIT and 18.40 % (V) for BHT less than 5 s after inspiration. The results of 149 subjects who had used the inhalation device showed that the VIE and EIT at 0 levels were significantly greater than those before training (Z= -5.651, -5.646, P < 0.001). The VIE and EIT at I-III and V significantly increased after training (all P < 0.05). CONCLUSIONS: Patients using portable inhaler devices do not always inhale with adequate flow patterns. The multiparametric inhalation assessment device may be useful in outpatient settings.

Performance testing for different peak expiratory flow meters.

BACKGROUND: Peak expiratory flow meters (PEFMs) have emerged as primary tools used for diagnosing and monitoring a range of respiratory diseases including asthma and chronic obstructive pulmonary disease, and the performance of these meters will thus impact disease evaluation. OBJECTIVE: The aim of this study was therefore to assess the technical performance of mechanical and electronic PEFMs commonly used in clinical practice. METHODS: The accuracy, repeatability, airflow resistance, frequency response, and linearity of five electronic and seven mechanical PEFMs were measured using a standard flow/volume simulator in accordance with nine A-waveforms and three B-waveforms defined in ISO 23747:2015 issued by the International Standards Organization (ISO). RESULTS: The accuracy, repeatability, linearity, airflow resistance, and frequency response pass rates for these 12 different PEFM brands were 41.67%, 75.00%, 50.00%, 75.00%, and 25.00%, respectively. Just 16.67% (2/12) of the tested PEFMs met all evaluated criteria, whereas the remaining PEFMs partially met these criteria. There were no significant differences between the two tested PEFM types in the low flow rate waveform test (P> 0.05), although there were significant differences in the medium and high flow rate waveform test (P< 0.05). In addition, the overall PEFMs test had poor accuracy and good repeatability, although most of the repeatability errors occurred in the BTPS state. CONCLUSION: PEFMs commonly used in clinical settings exhibit variable technical performance, and relevant departments need to strengthen PEFM quality control and management in China.

Remediation of soil contaminated with tris-(1-chloro-2-propyl) phosphate using plant and microorganism combined technology: A greenhouse experiment.

The remediation of tris-(1-chloro-2-propyl) phosphate (TCIPP) -contaminated soil by the plant (ryegrass, Lolium perenne L.) and microorganism (TCIPP degrading bacteria, Ochrobactrum sp. DT-6) alone or in combination was investigated in this study. TCIPP can inhibit the growth and development of ryegrass and there is a clear dose-effect relationship. Inoculation with strain DT-6 was able to mitigate the toxic influence of TCIPP on ryegrass, but this mitigation effect was not significant. TCIPP in the soil was relatively easy to be uptaken by the ryegrass roots and migrated to the shoots. Furthermore, as the soil TCIPP concentration rose, the concentration of TCIPP in ryegrass also exhibited a corresponding increase. The biological concentration factor (BCF) ranged from 0.33 to 1.88 and the biological accumulation coefficient (BAC) ranged from 0.54 to 3.98. They all significantly decreased with higher soil TCIPP concentrations. The translocation factor (TF) values ranged from 1.55 to 2.34. Inoculation of strain DT-6 significantly reduced TCIPP concentrations in ryegrass roots, stems, and leaves as well as the values of BAC and BCF under low and medium TCIPP concentration treatment conditions, but the effect on TF values was not remarkable. The planting of ryegrass significantly raised the elimination of TCIPP from the soil to 64.6-93.3%, but the influence of inoculation with strain DT-6 on the remediation effect by ryegrass was not significant. The percentage contribution of phytoextraction to the elimination of TCIPP from soils ranged from only 0.64-5.23%.

Predicting the Risk of Incorrect Inhalation Technique in Patients with Chronic Airway Diseases by a New Predictive Nomogram.

Purpose: To develop and internally validate a nomogram for predicting the risk of incorrect inhalation techniques in patients with chronic airway diseases. Methods: A total of 206 patients with chronic airway diseases treated with inhaled medications were recruited in this study. Patients were divided into correct (n=129) and incorrect (n=77) cohorts based on their mastery of inhalation devices, which were assessed by medical professionals. Data were collected on the basis of questionnaires and medical records. The least absolute shrinkage and selection operator method (LASSO) and multivariate logistic regression analyses were conducted to identify the risk factors of incorrect inhalation techniques. Then, calibration curve, Harrell's C-index, area under the receiver operating characteristic curve (AUC), decision curve analysis (DCA) and bootstrapping validation were applied to assess the apparent performance, clinical validity and internal validation of the predicting model, respectively. Results: Seven risk factors including age, education level, drug cognition, self-evaluation of curative effect, inhalation device use instruction before treatment, post-instruction evaluation and evaluation at return visit were finally determined as the predictors of the nomogram prediction model. The ROC curve obtained by this model showed that the AUC was 0.814, with a sensitivity of 0.78 and specificity of 0.75. In addition, the C-index was 0.814, with a Z value of 10.31 (P<0.001). It was confirmed to be 0.783 by bootstrapping validation, indicating that the model had good discrimination and calibration. Furthermore, analysis of DCA showed that the nomogram had good clinical validity. Conclusion: The application of the developed nomogram to predict the risk of incorrect inhalation techniques during follow-up visits is feasible.

Transcriptomic Analysis on Pectoral Muscle of European Meat Pigeons and Shiqi Pigeons during Embryonic Development.

In avian muscle development, embryonic muscle development determines the number of myofibers after birth. Therefore, in this study, we investigated the phenotypic differences and the molecular mechanism of pectoral muscle development of the European meat pigeon Mimas strain (later called European meat pigeon) and Shiqi pigeon on embryonic day 6 (E6), day 10 (E10), day 14 (E14) and day 1 after birth (P1). The results showed that the myofiber density of the Shiqi pigeon was significantly higher than that of the European meat pigeon on E6, and myofibers with a diameter in the range of 50~100 µm of the Shiqi pigeon on P1 were significantly higher than those of European meat pigeon. A total of 204 differential expressed genes (DEGs) were obtained from RNA-seq analysis in comparison between pigeon breeds at the same stage. DEGs related to muscle development were found to significantly enrich the cellular amino acid catabolism, carboxylic acid catabolism, extracellular matrix receptor interaction, REDOX enzyme activity, calcium signaling pathway, ECM receptor interaction, PPAR signaling pathway and other pathways. Using Cytoscape software to create mutual mapping, we identified 33 candidate genes. RT-qPCR was performed to verify the 8 DEGs selected-DES, MYOD, MYF6, PTGS1, MYF5, MYH1, MSTN and PPARG-and the results were consistent with RNA-seq. This study provides basic data for revealing the distinct embryonic development mechanism of pectoral muscle between European meat pigeons and Shiqi pigeons.

Simultaneous determination of six toxic alkaloids in human plasma and urine using capillary zone electrophoresis coupled to time-of-flight mass spectrometry.

A novel capillary zone electrophoresis separation coupled to electro spray ionization time-of-flight mass spectrometry method was developed for the simultaneous analysis of six toxic alkaloids: brucine, strychnine, atropine sulfate, anisodamine hydrobromide, scopolamine hydrobromide and anisodine hydrobromide in human plasma and urine. To obtain optimal sensitivity, a solid-phase extraction method using Oasis MCX cartridges (1 mL, 30 mg; Waters, USA) for the pretreatment of samples was used. All compounds were separated by capillary zone electrophoresis at 25 kV within 12 min in an uncoated fused-silica capillary of 75 µm id × 100 cm and were detected by time-of-flight mass spectrometry. This method was validated with regard to precision, accuracy, sensitivity, linear range, limit of detection (LOD), and limit of quantification (LOQ). In the plasma and urine samples, the linear calibration curves were obtained over the range of 0.50-100 ng/mL. The LOD and LOQ were 0.2-0.5 ng/mL and 0.5-1.0 ng/mL, respectively. The intra- and interday precision was better than 12% and 13%, respectively. Electrophoretic peaks could be identified by mass analysis.

An imputation-based genome-wide association study for growth and fatness traits in Sujiang pigs.

Sujiang pigs are a synthetic breed derived from Jiangquhai, Fengjing, and Duroc pigs. In this study, we sequenced the genome of 62 pigs with a coverage depth of 10× to 20×, including 27 Sujiang and 35 founder breed pigs, and we collected 360 global pigs' genome sequence data from public databases including 39 Duroc pigs. We obtained a high-quality variant dataset of 365 Sujiang pigs by imputing the porcine 80 K single nucleotide polymorphism (SNP) Beadchip to the whole-genome scale with a total of 422 pigs as a reference panel. A dataset of 365 imputated Sujiang pigs was used to perform single-trait genome-wide association study (GWAS) and meta-analyses for growth and fatness traits. Single-trait GWAS identified 1 907, 18, and 14 SNPs surpassing the suggestively significant threshold for backfat thickness, chest circumference, and chest width, respectively. Meta-analyses identified 2 400 genome-wide significant SNPs and 520 suggestively significant SNPs for backfat thickness and chest circumference, and 719 genome-wide significant SNPs and 1 225 suggestively significant SNPs for all seven traits. According to the meta-analysis of backfat thickness and chest circumference, a remarkable region of 2.69 Mb on Sus scrofa chromosome 4 containing FAM110B, IMPAD1, LYN, MOS, PENK, PLAG1, SDR16C5 and XKR4 was identified as a candidate region. The haplotype heat map of the 2.69 Mb region verified that Sujiang pigs were derived from Duroc and Chinese indigenous pigs, especially Jiangquhai pigs. The Kruskal-Wallis test showed that haplotypes of the 2.69 Mb region significantly affected backfat thickness and chest circumference traits. We then focused on PLAG1, an important growth-related gene, and identified two synonymous SNPs with obvious differences among different breeds in the PLAG1 gene. We then performed genotyping of 365 Sujiang, 150 Duroc, 95 Jiangquhai, and 100 Fengjing pigs to confirm the above result and verified that the two variants significantly affected phenotypes of growth and fatness traits. Our findings not only provide insights into the genetic architecture of porcine growth and fatness traits but also provide potential markers for selective breeding of these traits in Sujiang pigs.