The relationship between red cell distribution width, serum calcium ratio, and in-hospital mortality among patients with acute respiratory failure: A retrospective cohort study of the MIMIC-IV database.

To investigate the impact of RDW/CA (the ratio of red cell distribution width to calcium) on in-hospital mortality in patients with acute respiratory failure (ARF). This retrospective cohort study analyzed the data of 6981 ARF patients from the Medical Information Mart for Intensive Care (MIMIC-IV) database 2.0. Critically ill participants between 2008 and 2019 at the Beth Israel Deaconess Medical Center in Boston. The primary outcome of interest was in-hospital mortality. A Cox proportional hazards regression model was used to determine whether the RDW/CA ratio independently correlated with in-hospital mortality. The Kaplan-Meier method was used to plot the survival curves of the RDW/CA. Subgroup analyses were performed to measure the mortality across various subgroups. After adjusting for potential covariates, we found that a higher RDW/CA was associated with an increased risk of in-hospital mortality (HR = 1.17, 95% CI: 1.01-1.35, P = .0365) in ARF patients. A nonlinear relationship was observed between RDW/CA and in-hospital mortality, with an inflection point of 1.97. When RDW/CA ≥ 1.97 was positively correlated with in-hospital mortality in patients with ARF (HR = 1.554, 95% CI: 1.183-2.042, P = .0015). The Kaplan-Meier curve indicated the higher survival rates for RDW/CA < 1.97 and the lower for RDW/CA ≥ 1.97 after adjustment for age, gender, body mass index, and ethnicity. RDW/CA is an independent predictor of in-hospital mortality in patients with ARF. Furthermore, a nonlinear relationship was observed between RDW/CA and in-hospital mortality in patients with ARF.

Effects of Acute and Developmental Exposure to Bisphenol S on Chinese Medaka (Oryzias sinensis).

Bisphenol S (BPS), one of the substitutes for bisphenol A (BPA), is widely used in various commodities. The BPS concentrations in surface water have gradually increased in recent years, making it a predominant bisphenol analogue in the aquatic environment and raising concerns about its health and ecological effects on aquatic organisms. For this study, we conducted a 96 h acute toxicity test and a 15-day developmental exposure test to assess the adverse effects of BPS exposure in Chinese medaka (Oryzias sinensis), a new local aquatic animal model. The results indicate that the acute exposure of Chinese medaka embryos to BPS led to relatively low toxicity. However, developmental exposure to BPS was found to cause developmental abnormalities, such as decreased hatching rate and body length, at 15 dpf. A transcriptome analysis showed that exposure to different concentrations of bisphenol S often induced different reactions. In summary, environmental concentrations of BPS can have adverse effects on the hatching and physical development of Chinese medaka, and further attention needs to be paid to the potential toxicity of environmental BPS.

Integrated Analysis of the Transcriptome and Microbial Diversity in the Intestine of Miniature Pig Obesity Model.

Obesity, a key contributor to metabolic disorders, necessitates an in-depth understanding of its pathogenesis and prerequisites for prevention. Guangxi Bama miniature pig (GBM) offers an apt model for obesity-related studies. In this research, we used transcriptomics and 16S rRNA gene sequencing to discern the differentially expressed genes (DEGs) within intestinal (jejunum, ileum, and colon) tissues and variations in microbial communities in intestinal contents of GBM subjected to normal diets (ND) and high-fat, high-carbohydrate diets (HFHCD). After a feeding duration of 26 weeks, the HFHCD-fed experimental group demonstrated notable increases in backfat thickness, BMI, abnormal blood glucose metabolism, and blood lipid levels alongside the escalated serum expression of pro-inflammatory factors and a marked decline in intestinal health status when compared to the ND group. Transcriptomic analysis revealed a total of 1669 DEGs, of which 27 had similar differences in three intestinal segments across different groups, including five immune related genes: COL6A6, CYP1A1, EIF2AK2, NMI, and LGALS3B. Further, we found significant changes in the microbiota composition, with a significant decrease in beneficial bacterial populations within the HFHCD group. Finally, the results of integrated analysis of microbial diversity with transcriptomics show a positive link between certain microbial abundance (Solibacillus, norank_f__Saccharimonadaceae, Candidatus_Saccharimonas, and unclassified_f__Butyricicoccaceae) and changes in gene expression (COL6A6 and NMI). Overall, HFHCD appears to co-contribute to the initiation and progression of obesity in GBM by aggravating inflammatory responses, disrupting immune homeostasis, and creating imbalances in intestinal flora.

Leptin Receptor Gln223Arg Polymorphism of Human Spermatozoa Associated with Male Infertility in a Chinese Population.

Background: Leptin (LEP) is believed to play a crucial role in male reproduction, while the molecular mechanisms through which LEP affects the male reproductive system are unclear. LEP acts by binding to a leptin receptor (LEPR) which mediates its physiological action, but there are only limited studies on the function of LEPR in human sperm. Purpose: This study aimed to determine the Gln223Arg polymorphisms of the LEPR gene in human spermatozoa and evaluate their possible relationship with semen variables. Methods: The study was performed on Chinese men: 115 healthy subjects and 108 patients with primary and 98 with secondary infertility. Semen samples were obtained from all patients, and semen variables were analyzed. The genotypic and allelic frequencies of Gln223Arg polymorphism in spermatozoa were determined by PCR and restriction fragment length polymorphism (RFLP) analyses. Statistical analyses were performed using the chi-square test, the Kruskal-Wallis test, and the Mann-Whitney test. Results: There were no significant differences in genotypic or allelic frequency distributions of Gln223Arg polymorphism among men with primary infertility, secondary infertility, and controls. Similarly, semen volume and sperm concentration did not differ with the different genotypes in all groups of men. The percentages of motile sperm for AA + AG genotypes in men with primary infertility (31.98%) were significantly lower than those in secondary infertility, and control men with GG genotypes were 34.41% and 59.36%, respectively. At the same time, the percentages of normal morphology sperm for AA + AG genotypes in men with primary infertility (2.93%) were significantly lower than those in secondary infertility and control men with GG genotypes 3.71% and 6.54%, respectively. Conclusion: This study reveals a possible association between the Gln223Arg polymorphism of the LEPR gene in spermatozoa affecting spermatozoal membrane integrity and having a direct role in sperm motility.

Transcriptome Analysis of the Developmental Effects of Bisphenol F Exposure in Chinese Medaka (Oryzias sinensis).

Bisphenol F (BPF) has been used in the syntheses of polymers, which are widely used in coatings, varnishes, adhesives, and other plastics. During the past decades, BPF contamination in the aquatic environment has dramatically increased due to its release from manmade products. Concerns have driven much attention to whether it may adversely impact aquatic lives or human beings. The present study performed an acute toxic exposure experiment and a 15 d developmental exposure of BPF at environmental concentrations (20, 200, and 2000 ng/L) using Chinese medaka (Oryzias sinensis). In the acute toxic exposure, the LC50 of BPF to Chinese medaka is 87.90 mg/L at 96 h. Developmental exposure induced a significant increase in the frequency of larvae with abnormalities in the 2000 ng/L BPF group compared to the control group. Transcriptomic analysis of the whole larvae revealed 565 up-regulated and 493 down-regulated genes in the 2000 ng/L BPF exposure group. Analysis of gene ontology and KEGG pathways enrichments indicated endocrine disorders to be associated with BPF-induced developmental toxicity. The present results suggest that BPF is developmentally toxic at 2000 ng/L concentration in Chinese medaka and causes endocrine-related aberrations in the transcriptional network of genes.

Effects of nitrogen addition on greenhouse gas fluxes during continuous freeze-thaw cycles in a cold temperate forest.

Both nitrogen (N) deposition and soil freeze-thaw cycles (FTCs) induce pulses of greenhouse gas (GHG) emissions in cold temperate zones due to changes in soil carbon (C) and nitrogen (N) turnover. However, the combined effects of N addition and FTCs on GHG fluxes have received little research attention, particularly in boreal forests. We conducted a laboratory incubation experiment using intact soil cores from Rhododendron dauricum-Larix dahurica plots to investigate the GHG flux response to these combined effects. We separated the soil samples into seven groups (no, low, medium, and high sodium nitrate addition and low, medium, and high ammonium chloride addition) and exposed each group to continuous FTC conditions. The N2O and CO2 emissions were eventually stimulated by the FTCs, while CH4 uptake was inhibited by FTCs but responded differently under different N addition treatments. All the treatments had substantially increased N2O emissions compared to the control. However, the soil respiration rate significantly increased only with medium sodium nitrate addition, and high levels of N addition (regardless of form) inhibited CH4 uptake. These findings demonstrate that FTCs and N addition (in various forms and levels) have considerable effects on GHG emissions in temperate forest ecosystems. Moreover, dissolved organic carbon (DOC), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and inorganic nitrogen in soil are potential factors that drive GHG emissions and are necessary considerations in predicting future feedback effects of GHG emissions on climate change.

Responses of soil enzyme activity to freeze-thaw alternation in Greater Khingan Mountains, China.

We examined the effects of freeze-thaw cycles (0, 1, 3, 5, 7, 15 cycles) on the activities of urease, invertase, and proteinase in the different layers of the soil under the four typical stands in the cold temperate zone, i.e., Pinus pumila stand, Rhododendron-Betula platyphylla stand, Rhododendron-Larix gmelinii stand, Ledum-Larix gmelinii stand, with the indoor freeze-thaw simulation culture method being used. The relationship between soil enzyme activity and multiple physicochemical variables was analyzed during freeze-thaw alternation. The results showed that the activity of soil urease was firstly increased and then inhibited during freeze-thaw alternation. After freeze-thaw, urease activity did not differ from that without freeze-thaw. Invertase activity was firstly inhibited and then increased during freeze-thaw alternation, and was significantly increased by 8.5%-40.3% after freeze-thaw. Proteinase activity was firstly increased and then inhibited during freeze-thaw alternation, and was significantly decreased by 13.8%-68.9% after freeze-thaw. After freezing and thawing, there was significant positive correlation between urease activity and ammonium nitrogen and soil water content in Ledum-L. gmelinii stand and P. pumila stand, respectively, and proteinase activity had significant negative correlation with inorganic nitrogen concentration in P. pumila stand, Rhododendron-B. platyphylla stand, and Ledum-L. gmelinii stand. Invertase activity had significant positive correlation with organic matter in Rhododendron-L. gmelinii stand and Ledum-L. gmelinii stand.

Comparative Serum Proteome Analysis Indicates a Negative Correlation between a Higher Immune Level and Feed Efficiency in Pigs.

Identifying and verifying appropriate biomarkers is instrumental in improving the prediction of early-stage pig production performance while reducing the cost of breeding and production. The main factor that affects the production cost and environmental protection cost of the pig industry is the feed efficiency of pigs. This study aimed to detect the differentially expressed proteins in the early blood index determination serum between high-feed efficiency and low-feed efficiency pigs and to provide a basis for further identification of biomarkers using the isobaric tandem mass tag and parallel reaction monitoring approach. In total, 350 (age, 90 ± 2 d; body weight, 41.20 ± 4.60 kg) purebred Yorkshire pigs were included in the study, and their serum samples were obtained during the early blood index determination. The pigs were then arranged based on their feed efficiency; 24 pigs with extreme phenotypes were grouped as high-feed efficiency and low-feed efficiency, with 12 pigs in each group. A total of 1364 proteins were found in the serum, and 137 of them showed differential expression between the groups with high- and low-feed efficiency, with 44 of them being upregulated and 93 being downregulated. PRM (parallel reaction monitoring) was used to verify 10 randomly chosen differentially expressed proteins. The proteins that were differentially expressed were shown to be involved in nine pathways, including the immune system, digestive system, human diseases, metabolism, cellular processing, and genetic information processing, according to the KEGG and GO analyses. Moreover, all of the proteins enriched in the immune system were downregulated in the high-feed efficiency pigs, suggesting that a higher immune level may not be conducive to improving feed efficiency in pigs. This study provides insights into the important feed efficiency proteins and pathways in pigs, promoting the further development of protein biomarkers for predicting and improving porcine feed efficiency.

The Relationship Between Mindfulness, Fatigue, and Perceived Symptoms Among Frontline Nurses Who Performed Nucleic Acid Sample Collection During the COVID-19 in China: A Cross-Sectional Study.

Objective: Given the immense stress faced by medical staff during the COVID-19 pandemic, this study aimed to evaluate the relationship between mindful attention awareness, fatigue, and perceived symptoms among frontline nurses who performed nucleic acid sample collection during the COVID-19 pandemic, to reduce their fatigue and help them cope with perceived uncomfortable symptoms. Methods: A convenience sampling method was used to survey nurses who travelled to Hainan for nucleic acid sampling in August 2022 using an online (WeChat) questionnaire. A total of 514 frontline nurses who performed nucleic acid tests completed the questionnaire. The questionnaire covered basic demographic information, Mindful Attention Awareness Scale (MAAS) ratings, and Fatigue Severity Scale (FSS) ratings. Spearman correlation analysis was used to separate the relationship between MASS and FSS, and univariate and multivariate factor analyses were used to explore the relevant influences contributing to the occurrence of fatigue. Results: A total of 514 individuals completed the survey,93.97% (n=483) were female, mean age was 31.15 ± 5.7, MASS score was 69.01 ± 13.53, and 296 (57.59%) nurses experienced symptoms of fatigue during the auxiliary period. Spearman correlation analysis showed that FSS was associated with MASS. Multifactorial analysis showed that sex, age, marital status, fertility status, years of work, adaptation to dietary habits, hidrorrhea, and MAAS scores affected the presence of fatigue symptoms among the medical staff in Hainan (P<0.05). Conclusion: The psychological status of frontline nurses undergoing nucleic acid testing during the pandemic was poor, and the appearance of fatigue symptoms could be effectively reduced by increasing levels of positive thinking among medical staff to help them cope with public health emergencies.

Leptin Levels in Serum or Semen and Its Association with Male Infertility: A Meta-Analysis with 1138 Cases.

Background: Leptin has an association with male infertility. However, only sporadic studies inconsistently reported the results. Aim and Objective. In this study, we aimed to perform a meta-analysis to investigate the relationship between leptin and male infertility. Methods: This study was performed based on published articles related to leptin and infertile males. PubMed, Web of Science, Google Scholar, Ovid + Cochrane Central Register of Controlled Trials, Wiley Online Library, Chinese CNKI, Chinese Chong Qing VIP, Chinese Wan Fang, and China Biology Medicine databases were searched to identify all relevant studies. All eligible works of literature were analyzed by the "meta" or "metan" command in STATA version 12.0 software. The standardized mean difference (SMD) of leptin concentration in serum or semen and 95% confidence intervals (CIs) were estimated for all studies. The heterogeneity was described with I2. The sources of heterogeneity were explored via metaregression, and stratified analyses, sensitivity analyses, and publication bias were performed. Results: Nineteen studies were included in the current meta-analysis, involving 1138 cases of infertile men and 756 controls. The SMD of leptin concentration in serum was 2.002 (95% CI: 1.086, 2.918), Z-test (z) z = 4.29; p < 0.001, and I2 was 97.3%, p < 0.001. The SMD of leptin concentration in semen was 3.274 (95% CI: 2.137, 4.411), z = 5.64; p < 0.001, and I2 was 98.2%, p < 0.001. Notably, serum follicle-stimulating hormone (FSH) was slightly higher in infertile men (SMD = 3.695, z = 2.33, p = 0.020, I2 = 98.8%, p < 0.001). Other hormones, such as luteinizing hormone (LH) and testosterone, were also slightly higher, but the results were not statistically significant. In addition, sperm count (SMD = -4.533, 95% CI: -6.565, -2.501) and sperm motility (SMD = -7.894, 95% CI: -10.616, -5.172) inversely correlated with leptin levels in infertile males. Sperm abnormal forms did not show a statistically significant SMD of -0.076 (95% CI: -3.410, 3.258). Conclusion: Leptin plays a potential role in association with male infertility. This study may effectively reveal the relationship between leptin together with other hormones and its association with male infertility. These results may also provide opinions on precautionary measures.

Male nurses' work performance: A cross sectional study.

BACKGROUND: Nursing performance is closely related to the success of hospital and patient outcome. Recently, the proportion of male nurses has gradually increased. However, we do not know how these male nurses perform in clinical work. The purpose of this study was to understand the job performance of male nurses in China and to identify their risk factors. METHODS: This is a cross-sectional study. We contacted all the 30 public tertiary hospitals in Hunan Province and 26 of them cooperated. All the 647 male nurses in these hospitals were given questionnaires face-to-face between April 7 and August 8, 2020. The questionnaire included demographic information and the Schwirian's Six Dimension Scale. We also collected the family attitude and the main reason for choosing nursing. We then performed descriptive analyses and liner regressions on the collected data. RESULTS: We obtained valid questionnaires from 599 individuals. The median age of these male nurses was 26 years old and the nursing age was 4 years. They were mainly distributed in intensive care unit (ICU) (36.12%), operating room (27.42%), and emergency department (23.08%). And the means of the total scores for work performance was 176.42 (standard deviation (SD) = 20.62). The result of the regression shows that length of service, relationship status, educational level, department, main reason for choosing nursing, and family attitude are all risk factors of male nurses' work performance. CONCLUSION: Chinese male nurses are younger and have shorter working years. They mainly work in departments with higher work intensity and greater pressure. In addition, we found that years of service, education, marital status, department and main reasons for choosing nursing as factors influencing the job performance of male nurses.

[Potential turnover differences of exogenous alanine in soils of typical forests in the Xiaoxing'an Mountains, China]. / å°å ´å®‰å²­å ¸åž‹æ£®æž—åœŸå£¤ä¸­å¤–æºä¸™æ°¨é ¸çš„æ½œåœ¨å‘¨è½¬å·®å¼‚.

Taking five typical forests (Betula platyphylla forest, Quercus mongolica forest, B. platyphylla + Pinus koraiensis + Acer mono mixed forest, P. koraiensis forest, and Larix olgensis forest) in Xiaoxing'an Mountains as test objects, we investigated the effects of forest types (broadleaved forest, mixed forest, and coniferous forest) and soil layers (0-10 cm, 10-20 cm, 20-30 cm) on the turnover of exogenous alanine. The results showed that the turnover of soil exogenous alanine varied across different forest types, with significant difference in time and profile. The turnover rate was the highest in the broadleaved forest, followed by the mixed forest, and the lowest in the coniferous forest, with the half-life being 2.6-4.2, 3.6-5.5, and 4.3-7.0 h, respectively. With increasing soil depth, the potential turnover rate of exogenous alanine decreased, the half-life became longer, and soil adsorption of alanine weakened. With the addition of exogenous alanine, the content of ammonium and nitrate significantly increased, with the former being increased by 83.8%-95.3%, 80.9%-94.6% and 73.7%-93.2%, and the latter being increased by 82.9%-94.7%, 82.3%-93.8% and 78.1%-92.5% in the broadleaved forest, mixed forest, and coniferous forest, respectively. The net ammoniation and nitrification rates increased sharply at first and then decreased slowly, which were both positive. Soil layer and forest type as well as their interaction significantly affected the turnover, half-life, ammoniation, and nitrification of exogenous alanine.

Genetic Diversity and Population Structures in Chinese Miniature Pigs Revealed by SINE Retrotransposon Insertion Polymorphisms, a New Type of Genetic Markers.

RIPs have been developed as effective genetic markers and popularly applied for genetic analysis in plants, but few reports are available for domestic animals. Here, we established 30 new molecular markers based on the SINE RIPs, and applied them for population genetic analysis in seven Chinese miniature pigs. The data revealed that the closed herd (BM-clo), inbreeding herd (BM-inb) of Bama miniature pigs were distinctly different from the BM-cov herds in the conservation farm, and other miniature pigs (Wuzhishan, Congjiang Xiang, Tibetan, and Mingguang small ear). These later five miniature pig breeds can further be classified into two clades based on a phylogenetic tree: one included BM-cov and Wuzhishan, the other included Congjiang Xiang, Tibetan, and Mingguang small ear, which was well-supported by structure analysis. The polymorphic information contents estimated by using SINE RIPs are lower than the predictions based on microsatellites. Overall, the genetic distances and breed-relationships between these populations revealed by 30 SINE RIPs generally agree with their evolutions and geographic distributions. We demonstrated the potential of SINE RIPs as new genetic markers for genetic monitoring and population structure analysis in pigs, which can even be extended to other livestock animals.

Development and Genome Sequencing of a Laboratory-Inbred Miniature Pig Facilitates Study of Human Diabetic Disease.

Pig has been proved to be a valuable large animal model used for research on diabetic disease. However, their translational value is limited given their distinct anatomy and physiology. For the last 30 years, we have been developing a laboratory Asian miniature pig inbred line (Bama miniature pig [BM]) from the primitive Bama xiang pig via long-term selective inbreeding. Here, we assembled a BM reference genome at full chromosome-scale resolution with a total length of 2.49 Gb. Comparative and evolutionary genomic analyses identified numerous variations between the BM and commercial pig (Duroc), particularly those in the genetic loci associated with the features advantageous to diabetes studies. Resequencing analyses revealed many differentiated gene loci associated with inbreeding and other selective forces. These together with transcriptome analyses of diabetic pig models provide a comprehensive genetic basis for resistance to diabetogenic environment, especially related to energy metabolism.

Comprehensive inbred variation discovery in Bama pigs using de novo assemblies.

The Bama mini-pig (BM pig) is an inbred strain of the Bama Xiang pig (BX pig) and an important animal model used for studying human diseases. The extremely long inbreeding period renders a clear distinction between the features of the BM and BX strains, such as in their metabolism and olfactory system. However, there is limited information about differences between BM and BX animals at the genomic level. In this study, we generated genome sequencing data and used the assembly-vs-assembly approach to evaluate the phenotypic variations caused by inbreeding in these strains. Moreover, we detected differential expression of mutant genes related to the phenotypes in BX and BM pigs. We sequenced the genome of the BX pig strain and performed a series of analyses to reveal the comprehensive inbred genetic variants between BX and BM pigs. Here, the 2.56-Gb draft genome assembly for the BX pig and an N50 contig length of approximately 11.87 kb is described, and an N50 scaffold length of approximately 99 kb and the variations in the BX pig genome were identified by comparison with the BM pig reference genome. There were 1,424,354 single nucleotide polymorphisms (SNPs), 2,961,891 insertions and deletions (indels), 13,772 structural variants (SVs), and 20,606 copy number variants (CNVs) identified in the BX genome. Functional annotation of SVs and CNVs showed that the genes (ADGRE2, GPR143, olfactory receptor 52B4-like, olfactory receptor 10H1-like and SHROOM2) with both SVs and CNVs were enriched in the most of all KEGG pathways and gene ontology (GO) terms of mutant genes. ADGRE2, GPR143 and SHROOM2 were both found to have significant higher expression levels in BX pigs than in BM pigs. In the contrary, the expressions of olfactory receptor 52B4-like and olfactory receptor 10H1-like were significant lower in BX pigs than in BM pigs. In conclusion, sequence analysis of the BX pig genome revealed that the genome structure of the two pig strains has considerable genomic variation that was caused by the long inbreeding period. Moreover, qRT-PCR analysis of the mutant genes displayed a significant distinction that may be associated with phenotypic differences between these pig strains.

iTRAQ and PRM-based quantitative proteomics in T2DM-susceptible and -tolerant models of Bama mini-pig.

Type 2 diabetes mellitus (T2DM) is a complex, multifactorial metabolic disease, and the number of patients with T2DM has continued to increase in recent years. Large-scale proteomic studies on animal models of T2DM are of great importance to understand the pathophysiology of T2DM. Therefore, in our study, Isobaric tags for relative and absolute quantification (iTRAQ) and Parallel reaction monitoring (PRM) were used for proteomic analysis of skeletal muscles from T2DM-susceptible and -tolerant Bama mini-pig models induced by a high-fat, high-sugar diet. In our proteomic analysis, a total of 1646 proteins and 13 differentially expressed proteins (DEPs) were identified by iTRAQ-mass spectrometry, and 6 differentially expressed proteins were validated by PRM. Gene Ontology (GO) analysis revealed that most DEPs were extracellular matrix (ECM) proteins and participated in several biological processes, such as negative regulation of JAK-STAT cascade, negative regulation of STAT cascade, roundabout signaling pathway and peptide cross-linking via chondroitin 4-sulfate glycosaminoglycan, and the molecular functions of roundabout binding, glycosaminoglycan binding, heparin binding, sulfur compound binding, collagen binding, and kinase inhibitor activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis results showed that the differentially expressed proteins were involved in 14 pathways, including human disease pathways, metabolic pathways, signal transduction pathways, signaling molecules and interaction pathways, and the cellular process pathways associated with phagosomes and focal adhesion. In conclusion, the proteomics based on iTRAQ and PRM in T2DM-susceptible and -tolerant Bama mini-pig models showed that changes in amino acid metabolism, inflammation-associated pathways and the impaired function and environment of extracellular matrix are risk factors associated with increased pathogenesis of T2DM in Bama mini-pig.

Comparative analysis on liver transcriptome profiles of different methods to establish type 2 diabetes mellitus models in Guangxi Bama mini-pig.

AIMS/INTRODUCTION: Streptozotocin (STZ) is a permanent diabetogenic compound and often used in animal diabetes modeling. The aim of this study is to compare the liver transcriptome of type 2 diabetes models (T2DM) in Guangxi Bama Mini-pig (GBM pig) induced by STZ or Non-STZ. RESEARCH DESIGN AND METHODS: 22 female GBM pigs were divided into 4 groups. Ctr group (4 pigs): standard diets; DM1_HH group (10 pigs): high fat and high carbohydrate diets; DM2_HS group (4 pigs): high fat and high carbohydrate diets + STZ; DM3_SH group (4 pigs): STZ + high fat and high carbohydrate diets. Fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG) and total cholesterol (TC) were measured monthly. Glucose disappearance rate was evaluated by intravenous glucose tolerance test (IVGTT). Three pigs liver samples as biological replicates in each group were used for transcriptome sequencing analysis. RESULTS: All pigs injected with STZ were identified as diabetic. But only 4 of 10 pigs in DM1_HH group met the diabetes model standard. The most severe insulin resistance was observed in DM2_HS group. The FBG of DM1_HH, DM2_HS, DM3_SH and Ctr group were 7.20 ±â€¯0.17, 14.13 ±â€¯0.45, 7.98 ±â€¯0.99 and 4.43 ±â€¯0.27 mmol/L, respectively; the FINS were 53.67 ±â€¯1.82, 33.38 ±â€¯2.32, 49.91 ±â€¯1.50 and 39.78 ±â€¯1.14 mU/L, respectively. Compared with DM1_HH group, liver transcriptome showed that 7 genes were up-regulated while 10 were down-regulated in DM2_HS, 16 genes were up-regulated while 14 were down-regulated in DM3_SH. There were 4 co-expressive genes in DM2_HSvsDM1_HH and DM3_SHvsDM1_HH, including cystein rich 61 gene (CYR61), ribosomal protein L15 gene (RPL15), regulator of G protein signaling 1 gene (RGS1) and LOC102166695 gene. CONCLUSION: STZ can rapidly induce T2DM model in GBM pig, but STZ caused abnormal expression of genes in the liver.

Scriptaid Upregulates Expression of Development-Related Genes, Inhibits Apoptosis, and Improves the Development of Somatic Cell Nuclear Transfer Mini-Pig Embryos.

The present study was undertaken to investigate the mechanisms by which Scriptaid treatment improves the developmental competence of somatic cell nuclear transfer (SCNT) mini-pig embryos in vitro. We found that treatment with 500 nmol/L Scriptaid for 15 hours significantly improved the development of mini-pig SCNT embryos. Compared with the control group, the blastocyst rate was higher (18.3% vs. 10.7%; p < 0.05). The acetylation level on H3K14 of the Scriptaid-treated group was higher compared with the control group in SCNT embryos at two-cell, four-cell, and blastocyst stages (p < 0.05). After Scriptaid treatment, histone deacetylase gene HDAC5 expression level was significantly decreased in four-cell embryos and blastocysts, while the expression levels of the embryos' development-related genes AKT, Oct4, and apoptosis inhibited gene PGC-1α were significantly increased in blastocysts (p < 0.05). The number of apoptotic cells per blastocyst in the Scriptaid-treated group was lower compared with the control group (p < 0.05). These results indicate that Scriptaid repressed HDCA5 gene expression, increased the acetylation level of H3K14, upregulated the expression of AKT, Oct4, and PGC-1α genes, improved embryos' development, and reduced apoptosis, which favors development of the SCNT mini-pig embryos to blastocysts.

Chemiluminescent Detection for Estimating Relative Copy Numbers of Porcine Endogenous Retrovirus Proviruses from Chinese Minipigs Based on Magnetic Nanoparticles.

Chinese Bama minipigs could be potential donors for the supply of xenografts because they are genetically stable, highly inbred, and inexpensive. However, porcine endogenous retrovirus (PERV) is commonly integrated in pig genomes and could cause a cross-species infection by xenotransplantation. For screening out the pigs with low copy numbers of PERV proviruses, we have developed a novel semiquantitative analysis approach based on magnetic nanoparticles (MNPs) and chemiluminescence (CL) for estimating relative copy numbers (RCNs) of PERV proviruses in Chinese Bama minipigs. The CL intensities of PERV proviruses and the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were respectively determined with this method, and the RCNs of PERV proviruses were calculated by the equation: RCN of PERV provirus = CL intensity of PERV provirus/CL intensity of GAPDH. The results showed that PERVs were integrated in the genomes of Bama minipigs at different copy numbers, and the copy numbers of PERV-C subtype were greatly low. Two Bama minipigs with low copy numbers of PERV proviruses were detected out and could be considered as xenograft donor candidates. Although only semiquantitation can be achieved, this approach has potential for screening out safe and suitable pig donors for xenotransplantation.

Milk fat globule is an alternative to mammary epithelial cells for gene expression analysis in buffalo.

Owing to the difficulty in obtaining mammary gland tissue from lactating animals, it is difficult to test the expression levels of genes in mammary gland. The aim of the current study was to identify if milk fat globule (MFG) in buffalo milk was an alternative to mammary gland (MG) and milk somatic cell (MSC) for gene expression analysis. Six buffalos in late lactation were selected to collect MFG and MSC, and then MG was obtained by surgery. MFG was stained with acridine orange to successfully visualise RNA and several cytoplasmic crescents in MFG. The total RNA in MFG was successfully isolated and the integrity was assessed by agarose gel electrophoresis. We analysed the cellular components in MFG, MG and MSC through testing the expression of cell-specific genes by qRT-PCR. The results showed that adipocyte-specific gene (AdipoQ) and leucocyte-specific genes (CD43, CSF1 and IL1α) in MFG were not detected, whereas epithelial cell marker genes (Keratin 8 and Keratin 18) in MFG were higher than in MSC and lower than in MG, fibroblast marker gene (vimentin) in MFG was significantly lower than in MG and MSC, milk protein genes (LALBA, BLG and CSN2) and milk fat synthesis-related genes (ACC, BTN1A1, FABP3 and FAS) in MFG were higher than in MG and MSC. In conclusion, the total RNA in MFG mainly derives from mammary epithelial cells and can be used to study the functional gene expression of mammary epithelial cells.