Genetic analysis of health traits and their associations with longevity, fertility, production, and conformation traits in Holstein cattle.

Health traits have high economic values in dairy cattle breeding, which can cause considerable financial loss through involuntary culling. In this study, fourteen health traits were analysed, including five composite health traits: reproductive disorders, udder health (UH), digestive disorders, metabolic disorders, locomotory diseases (LD), and nine independent health traits: gestation disorders and peripartum disorders, irregular estrus cycle and sterility, metritis (ME), mastitis (MA), abomasal displacement (AD), enteritis (EN), and ketosis, claw diseases (CD), laminitis complex. This study analysed variance components for health traits through both single and bivariate repeatability animal models. All health traits showed low heritability, ranging from 0.001 to 0.025. Most of the health traits in five categories showed negative genetic correlations, ranging from -0.012 (CD and EN) to -0.634 (ME and EN). Strong positive genetic correlations appeared within the same category, ranging from 0.469 (EN and AD) to 0.994 (UH and MA, LD and CD). Furthermore, approximate genetic correlations were evaluated between health traits and routinely collected traits (longevity, fertility, production, and conformation). In general, the low to moderate approximate genetic correlations were estimated between health traits and routinely collected traits. The estimated correlations between health traits and longevity, fertility, production, and conformation traits could provide an indirect reference for disease-resistance breeding in Holstein cattle.

Genetic analysis of longevity and their associations with fertility traits in Holstein cattle.

The increase of longevity is intended to reduce involuntary culling rates, not extend the life span, and it reflects the ability of animals to successfully cope with the environment and disease during production. Sire model, animal model and repeatability animal models were used to estimate the (co) variance components of longevity and fertility traits. Six longevity and thirteen fertility traits were analysed, including herd life (HL), productive life (PL), number of days between first calving and the end of first lactation or culling (L1); number of days between first calving and the end of the second lactation or culling (L2); number of days between first calving and the end of the third lactation or culling (L3); number of days between first calving and the end of the fourth lactation or culling (L4); age at first service, age at first calving (AFC), the interval from first to last inseminations in heifer (IFLh), conception rate of first insemination in heifer, days open (DO), calving interval, gestation length, interval from calving to first insemination (ICF), interval from first to last inseminations in cow (IFLc), conception rate of first insemination in cow, calving ease (CE), birth weight, and calf survival. The estimated heritabilities (±SE) were 0.018 (±0.003), 0.015 (±0.003), 0.049 (±0.004), 0.025 (±0.003), 0.009 (±0.002) and 0.011 (±0.002) for HL, PL, L1, L2, L3 and L4, respectively. Strong correlations were appeared in HL and PL; the genetic and phenotypic correlation coefficients were 0.998 and 0.985, respectively. There were high genetic and phenotypic correlations which were observed in L1 and L2, L2 and L3, L3 and L4, respectively. All fertility traits of heifer showed medium to high heritability, while the cow showed low heritability. All heifer fertility traits had low genetic associations with longevity traits, ranging from -0.018 (L2 and IFLh) to 0.257 (L3 and AFC). Most of the fertility traits showed negative correlations with longevity traits in different parities, and we recommend DO, ICF, IFLc and CE as indirect indicators of longevity traits in dairy cows, but we also need to take into account the differences between parities.

miRNA-mRNA associations with inosine monophosphate specific deposition in the muscle of Jingyuan chicken.

1. Inosine monophosphate (IMP), is an essential component for meat flavour and microRNAs (miRNAs) play a vital role in its post-transcriptional regulation. However, the mechanism of how miRNA expression affects muscle-specific IMP deposition is unclear.2. The following study performed transcriptome sequencing and bioinformatics analysis of breast and leg muscle, which have significantly different IMP content in Jingyuan chicken. The differential miRNA-mRNAs were screened out and correlation analysis with IMP content was performed.3. A total of 39 differentially expressed miRNAs (DE miRNAs) and 666 differentially expressed mRNAs (DE mRNAs) were identified between breast muscles and leg muscles. Using miRNA-mRNA integrated analysis, 29 miRNA-target gene pairs were obtained, composed of 13 DE miRNAs and 28 DE mRNAs. Next, purine metabolism, glycolysis/gluconeogenesis, pyruvate metabolism and the biosynthesis of amino acid pathways as necessary for muscle IMP-specific deposition were identified. The differentially expressed gene PKM2, which was significantly enriched in all four pathways, is involved in IMP anabolism in the form of energy metabolism and enzyme activity regulation. The correlation analysis suggested that the gga-miR-107-3p-KLHDC2 negative interaction may be a key regulator in IMP deposition.4. This study explores the functional mechanism of IMP-specific deposition in Jingyuan chicken muscles at the miRNA and mRNA levels and highlights multiple candidate miRNAs and mRNAs for molecular-assisted breeding.

LncRNA LINC00958 promotes tumor progression through miR-4306/CEMIP axis in osteosarcoma.

OBJECTIVE: To investigate the mechanism by which LINC00958 affects osteosarcoma progression through miR-4306/CEMIP axis. PATIENTS AND METHODS: The microarray data (GSE66673) for gene expression in osteosarcoma cells were obtained from the Gene Expression Omnibus (GEO) database, and differentially expressed genes were analyzed by bioinformatics tools. Real-time quantitative PCR (RT-qPCR) was performed to detect the expression levels of LINC00958, miR-4306, and CEMIP in osteosarcoma tissues and cell lines. Western blot was performed to detect the expression levels of CEMIP. Subcellular fractionation analysis and RNA Fluorescence in situ hybridization (FISH) assay were performed to analyze the subcellular localization of LINC00958. The target relationship between LINC00958, CEMIP, and miR-4306 was verified by public bioinformatics database and dual-luciferase reporter assay. RNA immunoprecipitation (RIP) assay was performed to detect LINC00958 and miR-4306 bound to AGO2. The biological functions of LINC00958 and miR-205 on proliferation, cell cycle, apoptosis, migration, and invasion of osteosarcoma cells were evaluated by gain-of-function and loss-of-function experiments. Tumorigenic and metastatic abilities of cells in vivo were detected by xenograft tumor experiments and tumor metastasis assays in nude mice. Correlation between miR-4306 and LINC00958 or CEMIP expression in osteosarcoma tissues was analyzed using Pearson correlation analysis. Kaplan-Meier analysis was performed to assess the relationship between LINC00958 expression and overall survival of osteosarcoma patients. RESULTS: LINC00958 expression levels significantly increased in osteosarcoma tissues while miR-4306 expression levels significantly decreased, and the expression of these two genes was negatively correlated. Subcellular fractionation analysis and RNA FISH assay demonstrated that LINC00958 was mainly localized in the cytoplasm. Dual-luciferase reporter gene assay verified that LINC00958 competitively bound miR-4306 and repressed its expression. Silencing of LINC00958 inhibited proliferation, cell cycle, metastasis, and invasion of osteosarcoma cells while inducing cellular apoptosis. The introduction of miR-4306 inhibitors reversed the tumor-suppressing effect of silencing LINC00958. miR-4306 binds to CEMIP and suppressed its expression. Xenograft tumor experiments and tumor metastasis assays in nude mice demonstrated that silencing LINC00958 inhibited osteosarcoma cells' growth and metastasis while inhibiting miR-4306 reversed this effect. Kaplan-Meier analysis showed that high expression of LINC00958 was significantly associated with poor prognosis of osteosarcoma patients. CONCLUSIONS: LINC0095 promotes tumorigenesis and metastasis in osteosarcoma by competitively inhibiting miR-4306 expression, leading to elevated expression of CEMIP.

[The application of intravoxel incoherent motion diffusion imaging and 3D arterial spin labeling perfusion imaging in the evaluation of acute cerebral infarction].

Objective: To investigate the clinical value of magnetic resonance (MR) intravoxel incoherent motion (IVIM) diffusion imaging and 3D pseudo continuous arterial spin labeling (3D-pcASL) perfusion imaging in the evaluation of acute cerebral infarction. Methods: MR images of 49 patients with unilateral acute cerebral infarction diagnosed and treated in Affiliated Yancheng Hospital of Southeast University Medical College from October 2015 to February 2019 were retrospectively analyzed. High signal infarction area (S(D)) on diffusion image slice with the biggest lesion level and abnormal perfusion area (S(CBF)) on the corresponding level were measured. The presence of ischemic penumbra (IP) was represented by S(CBF)> S(D), and patients were divided into group IP and group non-IP. Regions of interest were set on the infarction core, brain tissue near the edge of the lesion (BNL) and their corresponding contralateral regions. The values of apparent diffusion coefficient (ADC), true diffusion coefficient (D), perfusion related diffusion coefficient (D(*)), perfusion fraction (f) and cerebral blood flow (CBF) of each region of interest were recorded and relative values of infarction lesion to its contralateral region (rADC, rD, rD(*), rf, rCBF) were calculated. Differences of each parameter value between infarction core, BNL and their corresponding contralateral regions and of each relative parameter value between infarction core and BNL, and between the two groups were compared.The differential diagnostic efficacy of relative parameter value with differences between groups was analyzed by receiver operating characteristics (ROC) curve. The correlations of each relative parameter value of 3D-pcASL and IVIM sequences were analyzed. Results: The ADC, D, f and CBF values of infarction core were significantly lower than those of contralateral regions in both groups (all P<0.01). Among all parameters of BNL in both groups, only the CBF value of group IP was significantly lower than that of contralateral region ((27.58±3.53) vs (41.20±5.66) ml·100 g(-1)·min(-1), P<0.01). The rADC, rD, rf and rCBF of infarction core were significantly lower than those of BNL in both groups (all P<0.01). The rCBF of BNL in group IP was significantly lower than that in group non-IP (0.68±0.12 vs 0.97±0.15, P<0.01), and the area under the curve was 0.949, the optimal threshold was 0.823, and the youden index was 0.855 for identifying the two groups. Other relative parameters values of infarction core and BNL had no statistical difference between the two groups. There were positive correlations between rCBF and rADC, rD, rf (r=0.428,0.335,0.565) of infarction core, rADC and rD, rf (r=0.853,0.602) of infarction core, also rADC and rD (r=0.336) of BNL (all P<0.05). Conclusions: IVIM can effectively evaluate the difusion and perfusion information of acute cerebral infarction lesions. However, its perfusion related parameters are not as good as 3D-pcASL in IP evaluation, which should be flexibly selected according to the actual needs of patients' condition evaluation.

[Inhibitory effect of DAPT on Notch signaling pathway in curcumin mediated photodynamic therapy for cervical cancer xenografts in nude mice].

Objective: Curcumin was used as photosensitizers in photodynamic therapy on cervical cancer xenografts in nude mice.Analysis the expression changes of Notch and downstream gene as NF-κB and VEGF before and after DAPT inhibition of Notch signaling pathway in vivo experiments.Our aim was to investigate the possible mechanism of Notch signaling pathway in the treatment of cervical cancer with PDT. Methods: A cervical cancer model of nude mice was established by subcutaneous inoculation of human cervical cancer Me180 cells 200 µl.After the success of the model, the experimental animals were divided into 4 groups (model group, curcumin PDT group, simple DAPT group, curcumin-PDT+ DAPT group), each group was 12.Tumor volume changes were analyzed and HE staining was observed in each group.MRNA and protein expression of Notch1 and its downstream NF-κB, VEGF were detected by RT-PCR, immunohistochemistry and Western blot before and after inhibition of Notch signaling pathway by DAPT. Results: Except the control group, the tumor volume of the other three groups remained unchanged or slightly reduced after 1-7 days of treatment.The difference was significant (P<0.05). HE staining showed the most obvious necrosis of curcumin-PDT group with DAPT.Both DAPT and curcumin-PDT could reduce the expression level of Notch1 in mRNA.The inhibition rates were 42.17% and 40.54%, respectively.And the inhibitory effect of curcumin-PDT with DAPT on Notch-1 was the strongest (79.22%) (P<0.01), and two of them had synergistic effect after combination with curcumin-PDT.But the expression of Notch-2 has no obvious inhibitory effect (P>0.05). Both DAPT and curcumin-PDT can inhibit the protein expression of Notch1, NF-κB and VEGF, and two of them have synergistic effect after combined use. Conclusions: DAPT can effectively block the Notch signaling pathway and inhibit the proliferation of cervical cancer cell line Me180.The application of DAPT to inhibit Notch signaling pathway after photodynamic therapy can achieve synergistic effect, which is mainly related to the down-regulation of the expression of Notch1 and NF-κB.Notch signaling pathway may be one of the targets of curcumin-PDT photodynamic therapy.

Analysis of gluconate metabolism for pyruvate production in engineered Escherichia coli based on genome-wide transcriptomes.

For pyruvate-producing strains, intracellular reduced nicotinamide adenine dinucleotide (NADH) accumulation is the main reason for the glycolysis inhibition. Comparing with glucose, using sodium gluconate as carbon source brought a decrease in NADH production and an increase in pyruvate production in engineered strain YP211. In order to explore the metabolic advantages of gluconate, genome-wide transcriptome analysis was employed to compare the metabolic differences between the two carbon sources. The results showed that the transcription of the genes gntU, gntK, and gntT responsible for transport and phosphorylation of gluconate, and genes edd and eda belonging to the Entner-Doudoroff (ED) pathway, was significantly enhanced. This suggested that the shortest route for the synthesis of pyruvate from gluconate was activated, and the synthesis of NADH was halved. Besides, the transcription of genes glpABCDTKF related to the glycerol metabolism was significantly enhanced, which might be because glycerol metabolism pathways were activated in the absence of glucose. These results provided valuable information for the further design of metabolic pathways in the construction of pyruvate-producing strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Comparing with glucose, using sodium gluconate as carbon source brought a decrease in nicotinamide adenine dinucleotide and an increase in pyruvate production in engineered strain YP211. From the genome-wide transcriptome analysis, the Entner-Doudoroff pathway was activated strongly in gluconate metabolism, which innovatively provided a shorter and more effective pathway for pyruvate production.

[Application of seven prediction models of vaginal birth after cesarean in a Chinese hospital].

OBJECTIVE: To evaluate the seven existing vaginal birth after cesarean (VBAC) screening tools and to identify additional factors that may predict VBAC or failed trial of labor in China. METHODS: In the study, 53 patients with 1 previous cesarean delivery who then delivered between January 1, 2007 and Novenber 31, 2014 were recruited. The average age of the patients was (32.1±3.5) years,the average gestational age was (38.0±2.3) weeks. There was no significant difference of the successful group and the failed group in the maternal/neonatal mortality and morbidity, also in the incidence of the postpartum hemorrhage and the postpartum infection. The probability of VBAC was calculated for each participant using 7 prediction models created by Weinstein, Flamm, Grobman, Gonen, Troyer, Smith and Torri. The data were analyzed using t test, rank-sum test, and receiver operating curve analysis. RESULTS: 44 trial of labor patients had a vaginal birth after cesarean delivery, and the successful rate was 83%. The scores between the successful group and the failed group had significant difference when evaluated by Weinstein and Grobman scoring models only. After recalculating the successful rate of VBAC in different score levels according to the references, there was significant difference between the rates of different score levels when evaluated by the Weinstein model. The successful rates of different score levels were higher compared to the references (<50%) when evaluated by the Troyer (70%), Gonen (60%), Torri (85.7%) models. The area under the receiver operating characteristic curve of Weinstein prediction model (0.746) and Flamm prediction model (0.723) were more than 0.7, and there was no significant difference between the seven models. CONCLUSION: Among the seven scoring models, the Weinstein model is more applicable to the population of our country, but a new model more applying to Chinese women still needs to be created.

Sequential projection pursuit with kernel matrix update and symbolic model selection.

This paper proposes a novel way for generating reliable low-dimensional features with improved class separability in a kernel-induced feature space. The feature projections rely on a very efficient sequential projection pursuit method, adapted to support nonlinear projections using a new kernel matrix update scheme. This enables the gradual removal of structure from the space of residual dimensions to allow the recovery of multiple projections. An adaptive kernel function is employed to unfold different types of data characteristics. We follow a holistic model selection procedure that, together with the optimal projections, dimensionality, and kernel parameters, additionally optimizes symbolically the projection index that controls the actual measurement of the data interestingness without user interaction. We tackle the underlying complex bi-level optimization model as a mixture of evolutionary and gradient search. The effectiveness of the proposed algorithm over existing approaches is demonstrated with benchmark evaluations and comparisons.

Ambient-temperature incubation for the field detection of Escherichia coli in drinking water.

AIMS: Escherichia coli is the pre-eminent microbiological indicator used to assess safety of drinking water globally. The cost and equipment requirements for processing samples by standard methods may limit the scale of water quality testing in technologically less developed countries and other resource-limited settings, however. We evaluate here the use of ambient-temperature incubation in detection of E. coli in drinking water samples as a potential cost-saving and convenience measure with applications in regions with high (>25°C) mean ambient temperatures. METHODS AND RESULTS: This study includes data from three separate water quality assessments: two in Cambodia and one in the Dominican Republic. Field samples of household drinking water were processed in duplicate by membrane filtration (Cambodia), Petrifilm™ (Cambodia) or Colilert® (Dominican Republic) on selective media at both standard incubation temperature (35­37°C) and ambient temperature, using up to three dilutions and three replicates at each dilution. Matched sample sets were well correlated with 80% of samples (n = 1037) within risk-based microbial count strata (E. coli CFU 100 ml−1 counts of <1, 1­10, 11­100, 101­1000, >1000), and a pooled coefficient of variation of 17% (95% CI 15­20%) for paired sample sets across all methods. CONCLUSIONS: These results suggest that ambient-temperature incubation of E. coli in at least some settings may yield sufficiently robust data for water safety monitoring where laboratory or incubator access is limited.

A case of hypoplastic left heart syndrome and bicuspid aortic valve in monochorionic twins.

The etiology of hypoplastic left heart syndrome (HLHS) remains unclear. Since a genetic cause for HLHS has not been obvious, it is generally considered to be inherited in a multifactorial manner. Studies of twins are valuable in elucidating the genetic contribution to a birth defect such as HLHS. We report a case of monochorionic twins in whom one has HLHS and the other has a bicuspid aortic valve. Predisposing genetic and environmental influences on individuals with identical genotypes, such as twins, may result in discordance of left-sided flow lesions.

Evidence for a role of endogenous nerve growth factor in the functional sex difference in the adrenergic innervation of the male and female mouse submandibular gland.

In view of the remarkable sex difference in nerve growth factor (NGF) content in mouse submandibular gland (SMG), a direct comparison of the adrenergic innervation as represented by noradrenaline (NA) content in SMG, rate of losing NA after superior cervical ganglionectomy, post-reserpine recovery of NA and survival and growth of implanted neurons of superior cervical ganglion into SMG was carried out. The results showed that the NA contents in the intact male SMG and in the male SMG during the recovery from reserpinization were higher than those in the female. There were marked sex differences in patterns of the time-course of post-ganglionectomy loss of NA in SMG and in the ability of SMG to support survival and growth of the implanted SCG neurons. These results indicate that NGF in the mouse SMG may play a functional role in the adrenergic neurons innervating the gland.