Massively Parallel CRISPR-Cas9 Knockout Screening in Sheep Granulosa Cells for FSH Response Genes.

Follicle-stimulating hormone (FSH) regulates ovarian follicle development through specific gene expression programs. Granulosa cells (GCs) are somatic cells surrounding the oocytes, secreting gonadotropins to regulate ovulation and promote follicular development. By analyzing the effects of different doses of FSH on the proliferation of GCs, we found that adding 10 ng/mL of FSH, as the optimal concentration, could promote the growth of GCs. Furthermore, we have successfully constructed the first CRISPR-Cas9 knockout library targeting the genes on chromosomes 2 and 3 and the X chromosomes of the sheep massively parallel coding gene, as well as an ovarian GCs knockout cell library. For the first time, we have exposed the knockout cell library to a concentration of 10 ng/mL FSH to explore the underlying mechanisms. Through this screening, we have identified 836 positive-negative screening genes that are responsive to FSH, thereby revealing the regulatory mechanisms and screening the functionality of candidate genes. Next, RNA-Seq of control (0 ng/mL), low (10 ng/mL), and high (100 ng/mL) doses of FSH revealed 1708 differentially expressed genes, and combined with 836 genes, we obtained 129 FSH dose-dependent genes with extremely significant differences. This enables us to delve deeper into investigating and identifying the mechanisms by which FSH regulates GCs. More generally, we have discovered new regulatory factors and identified reproductivity-associated major effectors. These findings provide novel research directions for further studies on sheep reproduction.

Chemical profile and quantitative comparison of constituents in different medicinal parts of Lactuca indica during varied harvest periods using UPLC-MS/MS method.

Lactuca indica L. cv. Mengzao (LIM), acknowledged as a pivotal "One Root of Medicine and Food", boasts dual applications in both culinary and medicinal domains. This research delves into the influence of various harvest periods (vegetative, budding, blossom, and fruiting) on distinct medicinal parts (roots, stems, leaves, flowers, and seeds) of LIM, employing plant metabolomics to assess its chemical constituents. A total of 66 chemical constituents were identified in LIM, with 11 chemical components emerging as potential markers for distinguish medicinal parts. Notably, nutritional organs exhibited elevated levels of cichoric acid, rutin and chlorogenic acid. Specifically, leaves during the budding stage displayed the highest chicoric acid content at 11.70 mg·g-1. Conversely, reproductive organs showed heightened concentrations of cichoric acid, rutin and chlorogenic acid, with seeds exhibiting the peak cichoric acid content at 4.53 mg g-1. This study enriches our understanding of LIM by offering novel insights into quality assessment and the comprehensive utilization of its diverse parts.

Development of clinical trials for non-small cell lung cancer drugs in China from 2005 to 2023.

Objective: Over the past few decades, the development of anti-cancer drugs in China has made outstanding achievements based on the support of national policies. To assess the progress of non-small cell lung cancer (NSCLC) drugs, we conducted a statistical analysis of clinical trials of drugs targeting NSCLC in China from 2005 to 2023. Methods: We downloaded, screened and analysed the data from three official websites, the Centre for Drug Evaluation of China National Medical Products Administration website (NMPA), ClinicalTrials.gov and the Chinese Clinical Trial Registry (ChiCTR). Results: From January 1, 2005 to April 15, 2023, a total of 1,357 drug clinical trials that met the standards were included, and the number of registered drug clinical trials has been increasing year by year, reaching the maximum of 199 in 2021. Among them, the maximum of 462 items (34.05%) in phase II clinical trials, followed by 333 (24.54%) in phase III clinical trials, and 139 (10.24%) in phase IV clinical trials. In all drug clinical trials, industry sponsored trials (ISTs) have 722 items (53.21%), which are higher than investigator-initiated trials (IITs). The clinical trials of chemical drugs have a maximum of 723 items (53.28%), while biopharmaceuticals have grown rapidly in the past 10 years, with a total of 374 (27.56%), and 48.19% of the drug clinical trials of combined medication. In addition, the geographical distribution of the leading units and participating units of Chinese drug clinical trials are uneven, and economic regions such as Beijing, Shanghai, Jiangsu are obviously ahead of other regions. Conclusion: From 2005 to 2023, the clinical trials of registered drugs for the treatment of NSCLC increased rapidly. Among them, due to the development of immunotherapy, the clinical trials of biopharmaceuticals and drugs for combined medication are growing most rapidly, while the exploration of the original drugs is a little far from enough. Our research provides a direction for the future drug clinical trials of NSCLC, laying foundation for further extending the survival rate of patients with NSCLC.

Characteristics and Mechanism of Acute Ischemic Stroke in NAVF Patients With Prior Oral Anticoagulant Therapy.

OBJECTIVES: We aimed to analyze the characteristics and mechanisms of acute ischemic stroke (AIS) in patients with nonvalvular atrial fibrillation (NVAF) who received prior anticoagulant therapy. METHODS: We retrospectively analyzed the data of patients with NVAF and AIS between January 2016 and December 2021. Patients were divided into non-anticoagulant, adequate anticoagulant, and insufficient anticoagulant groups according to their prior anticoagulant status. Patients with prior anticoagulant therapy were further divided into warfarin and direct oral anticoagulant groups. RESULTS: A total of 749 patients (661 without anticoagulants, 33 with adequate anticoagulants, and 55 with insufficient anticoagulants) were included. Patients with adequate anticoagulant had a milder National Institute of Health Stroke Scale at presentation ( P =0.001) and discharge ( P =0.003), a higher proportion of Modified Rankin Scale (mRS) ≤2 at discharge ( P =0.011), and lower rates of massive infarction ( P =0.008) than patients without anticoagulant. Compared with the non-anticoagulant group, the proportion of intravenous thrombolysis was significantly lower in the adequate anticoagulant ( P <0.001) and insufficient anticoagulant ( P =0.009) groups. Patients in the adequate anticoagulant group had higher rates of responsible cerebral atherosclerotic stenosis ( P =0.001 and 0.006, respectively) and competing large artery atherosclerotic mechanisms ( P =0.006 and 0.009, respectively) than those in the other 2 groups. Compared with warfarin, direct oral anticoagulant was associated with higher rates of Modified Rankin Scale ≤2 at discharge ( P =0.003). CONCLUSIONS: Adequate anticoagulant therapy may be associated with milder stroke severity and better outcomes at discharge in patients with NVAF. Competing large artery atherosclerotic mechanisms may be associated with anticoagulant failure in patients with NAVF with prior adequate anticoagulant therapy.

An evaluation of the qualitative superiority of the Mongolian medicinal herb hurdan-tsagaan (Platycodi Radix) from five different geographic origins based on anti-inflammatory effects.

ETHNOPHARMACOLOGICAL RELEVANCE: The contents and types of the active compounds in medicinal herbs depend greatly on their extraction methods, sources of origin and the modes of cultivation. Platycodon grandiflorus (Jacq.) A.DC. is an ethnic medicinal herb widely cultivated in China, and its dried root, Platycodi Radix (PR), is an important ingredient in herbal formulae for attenuating lung issues in Mongolian medical practice. However, research evaluating the superiority of PR based on harvesting regions is relatively limited. AIM: This study aimed to evaluate the qualitative superiority of PR from different regions based on anti-inflammatory effect. MATERIALS AND METHODS: A total of three commercial PR samples were obtained from Anguo, Bozhou and Shangluo, and two wild samples were obtained from Chifeng and Hinggan. PR extract (PRE) was prepared by water distillation, and platycodin D content in the extract was examined by HPLC-UVD. An optimal dose of PRE was administered to BALB/c mice with S. pneumoniae pneumonia, and IL-10 and TNF-α levels in lung tissue were examined by ELISA. HepG2 cells were treated with PRE, and an analysis of differentially expressed gene and functional enrichment was performed using an HTS2 assay. RESULTS: The contents of moisture, total ash, crude extract and platycodin D in the raw roots met the quality control requirements outlined in the Chinese Pharmacopoeia (2020 edition). The platycodin D content in the aqueous extract of the roots in descending order was 24.16% in PRE_Shangluo, 22.91% in PRE_Hinggan, 21.41% in PRE_Bozhou, 17.8% in PRE_Chifeng and 15.92% in PRE_Anguo. Furthermore, administration of PREs at an optimal dose of 2.0 g/kg resulted in some anti-inflammatory effect in mice with Streptococcus pneumoniae pneumonia, among which PRE_Shangluo administration exhibited a more obvious anti-inflammatory impact as shown by a significant decrease in the plasma white cell count (p < 0.05) and IL-10 level elevation and TNF-α reduction in lung tissue (p < 0.05) after treatment. In HepG2 cells treated with 100 µg/ml of each PRE, PRE_Hinggan and PRE_Shangluo resulted in significant differential expression of genes such as nuclear factor kappa B subunit 1 (NFKB1) and significant enrichment of pathways involved in the immune system, such as PI3K-Akt, MAPK and NF-kappa B signaling pathways. CONCLUSIONS: In this study, based on the anti-inflammatory effect, the quality of PR of Shangluo origin was superior to that of PR from the other four regions.

A novel "five-in-one" comprehensive medical care framework for rehabilitation and nursing.

With the evolution of society, the world has entered a moderate stage of aging. Not surprisingly, the aging problem in the world is getting more intense, resulting in the increasing demand for higher-quality and well-organized medical and elderly care services. To cope with that, many researchers have dedicated themselves to advancing the medical care system based on data or platforms. However, they have ignored the life cycle, health service and management and the inevitable shift of living scenarios for the elderly. Therefore, the study aims to improve health conditions and enhance senior citizens' life quality and happiness index. In this paper, we build a unified body for people in their old age, bridging the disconnection between medical care and elderly care and constructing the "five-in-one" comprehensive medical care framework. It should be mentioned that the system takes the human life cycle as its axis, relies on the supply side and supply chain management, integrates medicine, industry, literature and science as methods, and takes health service management as a requirement. Furthermore, a case study on upper limb rehabilitation is elaborated along the "five-in-one" comprehensive medical care framework to confirm the effectiveness of the novel system.

Recent advancements in digital health management using multi-modal signal monitoring.

Healthcare is the method of keeping or enhancing physical and mental well-being with its aid of illness and injury prevention, diagnosis, and treatment. The majority of conventional healthcare practices involve manual management and upkeep of client demographic information, case histories, diagnoses, medications, invoicing, and drug stock upkeep, which can result in human errors that have an impact on clients. By linking all the essential parameter monitoring equipment through a network with a decision-support system, digital health management based on Internet of Things (IoT) eliminates human errors and aids the doctor in making more accurate and timely diagnoses. The term "Internet of Medical Things" (IoMT) refers to medical devices that have the ability to communicate data over a network without requiring human-to-human or human-to-computer interaction. Meanwhile, more effective monitoring gadgets have been made due to the technology advancements, and these devices can typically record a few physiological signals simultaneously, including the electrocardiogram (ECG) signal, the electroglottography (EGG) signal, the electroencephalogram (EEG) signal, and the electrooculogram (EOG) signal. Yet, there has not been much research on the connection between digital health management and multi-modal signal monitoring. To bridge the gap, this article reviews the latest advancements in digital health management using multi-modal signal monitoring. Specifically, three digital health processes, namely, lower-limb data collection, statistical analysis of lower-limb data, and lower-limb rehabilitation via digital health management, are covered in this article, with the aim to fully review the current application of digital health technology in lower-limb symptom recovery.

A 1.1 Mb duplication CNV on chromosome 17 contributes to skeletal muscle development in Boer goats.

The Boer goat is one of the top meat breeds in modern animal husbandry and has attracted widespread attention for its unique growth performance. However, the genetic basis of muscle development in the Boer goat remains obscure. In this study, we identified specific structural variants in the Boer goat based on genome-wide selection signals and analyzed the basis of the molecular heredity of related candidate genes in muscle development. A total of 9 959 autosomal copy number variations (CNVs) were identified through selection signal analysis in 127 goat genomes. Specifically, we confirmed that the highest signal CNV (HSV) was a chromosomal arrangement containing an approximately 1.11 Mb (CHIR17: 60062304-61171840 bp) duplicated fragment inserted in reverse orientation and a 5 362 bp deleted region (CHIR17:60145940-60151302 bp) with overlapping genes (e.g., ARHGAP10, NR3C2, EDNRA, PRMT9, and TMEM184C). The homozygous duplicated HSV genotype (+/+) was found in 96% of Boer goats but was not detected in Eurasian goats and was only detected in 4% of indigenous African goats. The expression network of three candidate genes ( ARHGAP10, NR3C2, and EDNRA) regulating dose transcription was constructed by RNA sequencing. Results indicated that these genes were involved in the proliferation and differentiation of skeletal muscle satellite cells (SMSCs) and their overexpression significantly increased the expression of SAA3. The HSV of the Boer goat contributed to superior skeletal muscle growth via the dose effects of overlapping genes.

Comparative analysis of the genetic diversity of the neutral microsatellite loci and second exon of the goat MHC-DQB1 gene.

This study compared and analyzed the genetic diversity and population structure of exon 2 of the DQB1 gene and 13 autosomal neutral microsatellite markers from 14 Chinese goat breeds to explore the potential evolutionary mechanism of the major histocompatibility complex (MHC). A total of 287 haplotypes were constructed from MHC-DQB1 exon 2 from 14 populations, and 82 nucleotide polymorphic sites (SNPs, 31.78%) and 172 heterozygous individuals (79.12%) were identified. The FST values of the microsatellites and MHC-DQB ranged between 0.01831-0.26907 and 0.00892-0.38871, respectively. Furthermore, 14 goat populations showed rich genetic diversity in the microsatellite loci and MHC-DQB1 exon 2. However, the population structure and phylogenetic relationship represented by the two markers were different. Positive selection and Tajima's D test results showed the occurrence of a diversified selection mechanism, which was primarily based on a positive and balancing selection in goat DQB. This study also found that the DQB sequences of bovines exhibited trans-species polymorphism (TSP) among species and families. In brief, this study indicated that positive and balancing selection played a major role in maintaining the genetic diversity of DQB, and TSP of MHC in bovines was common, which enhanced the understanding of the MHC evolution.

Identification of differentially expressed long non-coding RNAs and messenger RNAs involved with muscle development in Dazu black goats through RNA sequencing.

This study aimed to explore the genetic basis of muscle development in goats. The transcriptome dataset for differentially expressed lncRNAs (DELs) and differentially expressed genes (DEGs) of goat muscle at different developmental stages were obtained using RNA-Seq. A total of 447,806,481 and 587,559,465 clean reads in the longissimus dorsi muscle of Dazu black goats between 75d embryonic stage and 1d after birth were generated through Illumina paired-end sequencing, and their mapping rates were 89.82 and 90.99%, respectively. Moreover, 4517 DEGs and 648 DELs were identified, and 4784 lncRNA-mRNA targeting relationships were predicted. Gene function annotation results showed that 4101 DEGs were significantly enriched to 1098 GO terms, and 2014 DEGs were significantly enriched to 40 KEGG pathways, including many GO terms and pathways related to muscle development, such as cell differentiation and Wnt signaling pathway. Then, 10 DELs and 20 DEGs were randomly selected for RT-qPCR verification, and the agreement rate between the verification and RNA-Seq results was 90%, indicating the high reliability of the RNA-Seq data analysis. In conclusion, this study obtained several mRNAs and lncRNAs related to the muscle development of Dazu black goats and identified several targeted regulatory pairs of lncRNA-mRNA. This study may serve as a reference to understand the genetic basis and molecular mechanism of muscle development in goats.

Bioinformatic Prediction of Depression-Related Signaling Pathways Regulated by miR-146a in Peripheral Blood of Patients with Post-Stroke Depression.

It was aimed to explore the differential expression of miR-146a-5p in peripheral blood of patients with post-stroke depression (PSD), and to analyze its mechanism using bioinformatics. Stroke patients were selected as the research objects, and were divided into PSD ones and non-post-stroke depression (N-PSD) ones with the National Institutes of Health stroke scale (NHISS) and Hamilton Depression Scale-17 terms (HAMD-17) scores. Peripheral blood of patients was collected for serum miR-146a-5p detection. Targetscan7.1, miRDB, DIANA TOOLS, and more databases were used to predict the target genes of miR-146a-5p. String11.0 was applied to construct a protein interaction network, and GO and KEGG pathway enrichment analysis of target genes was performed. Compared with that of N-PSD patients, serum miR-146a-5p levels in PSD patients were significantly increased (P<0.05). The receiver operator characteristic (ROC) curve suggested that the sensitivity and specificity of miR-146a-5p in predicting PSD were 0.703 and 0.811, respectively. The human miR-146a-5p sequence was highly conserved, with a total of 43 target genes. It involved analysis of activity, signaling pathways, and transcriptional regulation, as well as related signaling pathways such as Toll-like receptors (TLR), neurotrophic factors, and nuclear factor kappa-B (NF-κB). In conclusion, the expression level of miR-146a-5p was abnormally increased in PSD patients, and it could be taken as a candidate marker for the diagnosis of PSD. miR-146a-5p could affect PSD through signaling pathways of TLRs, neurotrophic factors, and NF-κB.

Streptococcus humanilactis sp.nov., isolated from healthy nursing mother's breast milk.

Two bacterial strains were isolated from the breast milk of two healthy nursing mothers. The isolates were Gram-positive, catalase-negative, coccus-shaped, chain-forming organisms. Analysis of the 16S rRNA gene sequences of strain IMAU99125T shared 99.7 and 99.6% similarity with Streptococcus mitis ATCC 49456 T and Streptococcus pseudopneumoniae ATCC BAA-960 T, respectively. The nearly complete 16S rRNA gene sequences of IMAU99125T and IMAU99674 strains were very closely related (with only 0.06% difference between them). Sequence analysis of the gyrB and rpoB genes also indicated that IMAU99125T was closely related to S. mitis ATCC 49456 T (94.7% and 97.1%, respectively) and S. pseudopneumoniae ATCC BAA-960 T (94.4% and 97.1%, respectively). Average nucleotide identity (ANI) values between strain IMAU99125T and S. mitis ATCC 49456 T and S. pseudopneumoniae ATCC BAA-960 T, were 93.3% and 92.7%, respectively. Genome-to-genome distance (GGD) values between strain IMAU99125T and S. mitis ATCC 99125 T and S. pseudopneumoniae ATCC BAA-960 T were 53.4% (50.7-56.0) and 50.4% (47.7-53.0), respectively. The major fatty acids of the strain were C16:0 (51.4%). On the basis of the results of phenotypic and phylogenetic analyses, we propose that the two strains be classified as representing a novel species of the genus Streptococcus, namely Streptococcus humanilactis sp.nov. The type strain is IMAU99125T (= GDMCC 1.1876 T = KCTC 21157 T). The genome of Streptococcus humanilactis sp. nov. is comprised of 2,027,143 bp. The DNA G + C content of the strain is 40.0 mol%.

Adoption of Compound Echocardiography under Artificial Intelligence Algorithm in Fetal Congenial Heart Disease Screening during Gestation.

This research was aimed at exploring the diagnostic and screening effect of composite echocardiography based on the artificial intelligence (AI) segmentation algorithm on fetal congenital heart disease (CHD) during pregnancy, so as to reduce the birth rate of newborns with CHD. A total of 204 fetuses with abnormal heart conditions were divided into group II, group C (optimized with the AI algorithm), and group W (not optimized with the AI algorithm). In addition, 9,453 fetuses with normal heart conditions were included in group I. The abnormal distribution of fetal heart and the difference of cardiac Z score between group II and group I were analyzed, and the diagnostic value of group C and group W for CHD was compared. The results showed that the segmentation details of the proposed algorithm were better than those of the convolutional neural network (CNN), and the Dice coefficient, precision, and recall values were higher than those of the CNN. In fetal CHD, the incidence of abnormal ultrasonic manifestations was ventricular septal defect (98/48.04%), abnormal right subclavian artery (29/14.22%), and persistent left superior vena cava (25/12.25%). The diagnostic sensitivity (75.0% vs. 51.5%), specificity (99.6% vs. 99.2%), accuracy (99.0% vs. 98.2%), negative predictive value (88.5% vs. 78.5%), and positive predictive value (99% vs. 57.7%) of echocardiography segmentation in group C were significantly higher than those in group W. To sum up, echocardiography segmented by the AI algorithm could obviously improve the diagnostic efficiency of fetal CHD during gestation. Cardiac ultrasound parameters of children with CHD changed greatly.

Root Carbon Resources Determine Survival and Growth of Young Trees Under Long Drought in Combination With Fertilization.

Current increases in not only the intensity and frequency but also the duration of drought events could affect the growth, physiology, and mortality of trees. We experimentally studied the effects of drought duration in combination with fertilization on leaf water potential, gas exchange, growth, tissue levels of non-structural carbohydrates (NSCs), tissue NSC consumption over-winter, and recovery after drought release in oak (Quercus petraea) and beech (Fagus sylvatica) saplings. Long drought duration (>1 month) decreased leaf water potential, photosynthesis, and NSC concentrations in both oak and beech saplings. Nitrogen fertilization did not mitigate the negative drought effects on both species. The photosynthesis and relative height increment recovered in the following rewetting year. Height growth in the rewetting year was significantly positively correlated with both pre- and post-winter root NSC levels. Root carbon reserve is critical for tree growth and survival under long-lasting drought. Our results indicate that beech is more sensitive to drought and fertilization than oak. The present study, in a physiological perspective, experimentally confirmed the view that the European beech, compared to oak, may be more strongly affected by future environmental changes.

Genome-Wide Selection Sweep between Wild and Local Pigs from Europe for the Investigation of the Hereditary Characteristics of Domestication in Sus Scrofa.

The phenotypic characteristics of existing domestic pigs (DPs) greatly differ from those of wild boar (WB) populations thousands of years ago. After thousands of years of human domestication, WB and DP have diverged greatly in terms of genetics. Theoretically, worldwide local pigs have independent contributions from their local WBs at the beginning of Sus scrofa domestication. The investigation of the vicissitude of the heredity material between domestic populations and their wild ancestors will help in further understanding the domestication history of domestic animals. In the present study, we performed a genome-wide association scan (GWSA) and phylogeny estimation with a total of 1098 public European Illumina 60K single nucleotide polymorphism data, which included 650 local DPs and 448 WBs. The results revealed that the phylogenetic relationship of WBs corresponds to their geographical distribution and carries large divergence with DPs, and all WB breeds (e.g., HRWB, SBWB, and TIWB) presents a closely linkage with the middle WB (e.g., HRWB, and PLWB). In addition, 64 selected candidate genes (e.g., IDH2, PIP5K1B, SMARCA2, KIF5C, and TJP2) were identified from GWSA. A total of 63 known multiple biological functional pathways were annotated by 22 genes, and ubiquinone and other terpenoid-quinone biosynthesis pathways that belong to the metabolism of cofactors and vitamins were significantly enriched (p < 0.05). The most frequent (28.57%) pathways were classified under metabolism. We confirmed that the middle European WB has made an important genetic contribution to the entire European WB populations. A series of selected genes discovered from this study provides the scientific community with a deeper understanding of the heredity performance of metabolism and emotion and the real purpose behind domestication.

Identification of differentially expressed microRNAs in ovulatory and subordinate follicles in Dazu black goats.

How multiple ovulations happen in prolific goats is still unknown even though studies on ovarian physiology and folliculogenesis have made extensive progress. MicroRNAs (miRNAs) are a class of endogenous regulatory factors and regulate structural gene expression mainly at the post-transcriptional level. In this study, in the follicular phase, six large follicles were collected from prolific Dazu black goat and used to generate RNA libraries for RNA sequencing. Based on the litter size and average number of ovulatory follicles in Dazu black goats, the largest three follicles were sorted as ovulatory follicles, and the remaining as subordinate ones. In total, 418 known miRNAs and 110 novel miRNAs were found, and the expression of six randomly selected miRNAs was validated by quantitative PCR analysis. Nine miRNAs were differently expressed between the ovulatory and subordinate follicles (p < 0.01). Chi-miR-582-5p, novel-130, chi-miR-214-3p, and chi-miR-500-5p were upregulated in the ovulatory group, and chi-miR-383, chi-miR-130b-5p, chi-miR-92a-3p, chi-miR-125b-5p, and novel-9 were downregulated in the same group. Chi-miR-130b-5p and chi-miR-214-3p were predicted to target at LHR (XM_013967581.1), GDF9 (NM_001285708.1), BMP15 (NM_001285588.1), and CYP19A1 (XM_013967046.1). In conclusion, nine miRNAs were differently expressed between ovulatory and subordinate follicles, and chi-miR-130b-5p and chi-miR-214-3p were predicted to regulate the expression of genes involved in gonadotropin hormone signaling and oocyte-derived growth factors. Additional studies are needed to confirm these findings.

A nonsynonymous SNP within the AMH gene is associated with litter size in Dazu black goats.

AMH, KISS1R and GDF9 genes play a vital role in human and animal reproduction and might be used as the genetic markers for the reproduction traits selection. The aim of this study was to screen the single nucleotide polymorphisms (SNPs) within the AMH, KISS1R and GDF9 genes and to determine the correlations between these SNPs and the litter size in goats. Nine single SNPs within these genes were used for genotyping of the 190 Dazu black goat populations by SNaPshot technique. The polymorphisms of nine SNPs within these genes were detected in Dazu black goats. The significant correlation was observed between one SNP (g.89172108A > C) within the AMH gene and the litter size of second born in Dazu black goats (p < 0.05). The SNP was located in exon 4 (XM_018050765.1) of the AMH gene and was one nonsynonymous substitution, which resulted in a change of an amino acid from Glutamine to Proline (Gln38Pro). These results suggested that the nonsynonymous SNP g.89172108A > C of AMH gene could be used as a potential genetic marker for Marker-assisted selection (MAS) in goats breeding programs.

Prognostic role of serum ammonia in patients with sepsis-associated encephalopathy without hepatic failure.

Objectives: Our previous study shows that serum ammonia in sepsis patients without hepatic failure is associated with a poor prognosis. The relationship between serum ammonia level and the prognosis of sepsis-associated encephalopathy (SAE) patients without hepatic failure remains unclear. We aimed to explore the relationship between serum ammonia levels and the prognosis of patients with SAE. Materials and methods: This study is a retrospective cohort study. We collected 465 patients with SAE admitted to the intensive care unit (ICU) from Medical Information Mart for Intensive Care IV (MIMIC IV) from 2008 to 2019. Patients with SAE were divided into a survival group (369 patients) and a non-survival group (96 patients). We used the Wilcoxon signed-rank test and the multivariate logistic regression analysis to analyze the relationship between serum ammonia levels and the prognosis of patients with SAE. R software was used to analyze the dataset. Results: The primary outcome was the relationship between serum ammonia level and hospital mortality of SAE. The secondary outcomes were the relationship between serum ammonia level and hospital stays, simplified acute physiology score (SAPS II), Charlson, Glasgow coma scale (GCS), sequential organ failure assessment (SOFA), and lactate level of SAE. The mortality of patients with SAE was 20.6%. The serum ammonia level was not significantly associated with hospital mortality, longer hospital stays, higher SAPS II and Charlson scores, and lower GCS of patients with SAE. The serum ammonia level was associated with higher SOFA scores and lactate levels in patients with SAE. The SAPS II and Charlson scores were independent risk factors for death in patients with SAE. Conclusion: Serum ammonia level was associated with higher SOFA scores and lactate levels in patients with SAE. In addition, the SAPS II and Charlson scores can be used to assess the prognosis of patients with SAE. Therefore, we should closely monitor serum ammonia, SAPS II, and Charlson levels in patients with SAE.

Genetic Basis of Follicle Development in Dazu Black Goat by Whole-Transcriptome Sequencing.

The follicle development (FD) is an important factor determining litter size in animals. Recent studies have found that noncoding RNAs (ncRNAs) play an important role in FD. In particular, the role of the regulatory mechanism of competing endogenous RNAs (ceRNAs) that drive FD has attracted increasing attention. Therefore, this study explored the genetic basis of goat FD by obtaining the complete follicular transcriptome of Dazu black goats at different developmental stages. Results revealed that 128 messenger RNAs (mRNAs), 4 long noncoding RNAs (lncRNAs), 49 microRNAs (miRNAs), and 290 circular RNAs (circRNAs) were significantly differentially expressed (DE) between large and small follicles. Moreover, DEmRNAs were enriched in many signaling pathways related to FD, as well as GO terms related to molecular binding and enzyme activity. Based on the analysis of the ceRNA network (CRN), 34 nodes (1 DElncRNAs, 10 DEcircRNAs, 14 DEmiRNAs, and 9 DEmRNAs) and 35 interactions (17 DEcircRNAs-DEmRNAs, 2 DElncRNAs-DEmiRNAs, and 16 DEmRNA-DEmiRNAs) implied that the CRN could be involved in the FD of goats. In conclusion, we described gene regulation by DERNAs and lncRNA/circRNA-miRNA-mRNA CRNs in the FD of goats. This study provided insights into the genetic basis of FD in precise transcriptional regulation.