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Aim: To discover novel methylation-related differentially expressed genes (MRDEGs) for cervical cancer, with a focus on their potential for early diagnosis and prognostic assessment. Materials & methods: We integrated data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. TCGA-MRDEGs were identified by analyzing differentially methylated genes (DMGs) and their correlation with gene expression. We examined GEO datasets GSE39001, GSE9750, and GSE46306 for GEO-MRDEGs. Overlapping MRDEGs were subjected to overall survival (OS) analysis to identify prognostic markers. The expression and methylation levels of these genes were validated in a total of 30 tissue samples, comprising 20 from cervical cancer patients and 10 from normal cervical tissues, using qRT-PCR and MassARRAY EpiTYPER Assay. Results: A total of 314 TCGA-MRDEGs and 40 GEO-MRDEGs were identified. Intersection analysis yielded 10 overlapping MRDEGs. Notably, NOVA1, GSTM5, TRHDE, and CXCL12 were found to have reduced expression and increased methylation in cervical cancer, which correlated with poor prognosis. The methylation status and expression levels of these genes were confirmed in tissue specimens. Conclusion: We identified four MRDEGs as potential prognostic biomarkers for cervical cancer. Their clinical utility is highlighted, but further validation in larger cohorts is required to establish their clinical significance.

To compare the impact of nanoselenium and sodium selenite on the performance, blood indices, and milk metabolites of dairy cows during the peak lactation period, two groups of dairy cows under the same conditions were selected as the control group (CON group) and treatment group (NSe group) for a 38-day (10 days for adaptation and 28 days for sampling) experiment. The control group (CON) was provided a basal diet +3.3 g/d of sodium selenite (purity1%), whereas the nanoselenium group (NSe) was offered the same diet +10 mL/d of nanoselenium (selenium concentration 1,500 mg/L). The results showed that NSe significantly increased the milk yield, milk selenium content, and feed efficiency (p 0.05). NSe significantly increased blood urea nitrogen (BUN) and alkaline phosphatase (ALP) (p 0.05). In addition, the nontargeted metabolomics of the milk was determined by LC–MS technology, and the differentially abundant metabolites and their enrichment pathways were screened. According to these findings, NSe considerably increased the contents of cetylmannoside, undecylenoic acid, 3-hydroxypentadecanoic acid, 16-hydroxypentadecanoic acid, threonic acid, etc., but decreased the contents of galactaric acid, mesaconic acid, CDP-glucose etc. Furthermore, the enriched metabolic pathways that were screened with an impact value greater than 0.1 included metabolism of niacin and niacinamide, pyruvate, citrate cycle, riboflavin, glycerophospholipid, butanoate and tyrosine. Pearson correlation analysis also revealed a relationship between different milk metabolites and blood selenium, as well as between milk selenium and blood biochemical indices. In conclusion, compared with sodium selenite, nanoselenium improves the milk yield, feed efficiency, and milk selenium content of dairy cows and regulates milk metabolites and related metabolic pathways in Holstein dairy cows during the peak lactation period, which has certain application prospects in dairy production.

The objective of this experiment was to investigate the effects of Lactobacillus plantarum and molasses on the nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora of alfalfa silage in sandy grasslands. The experimental treatments included control (CK), 106 CFU/g Lactobacillus plantarum (L), 5% molasses (M), and 106 CFU/g Lactobacillus plantarum + 5% molasses (LM). The nutrient composition, fermentation quality, bacterial count, aerobic stability, and microflora were determined after 14 days and 56 days of ensiling, respectively. The results showed that the addition of L, M, and LM reduced dry matter loss (DM), neutral detergent fiber (NDF), and acid detergent fiber (ADF) content, and increased water-soluble carbohydrates (WSC) and ether extract (EE) content, compared to the CK group. Meanwhile, more lactic acid (LA) and accelerated fermentation were observed, causing the pH value to drop below 4.5 in the L, M, and LM groups after 56 days of ensiling. The addition of L, M, and LM promoted lactic acid bacteria (LAB), and inhibited yeast. The addition of L significantly increased the content of acetic acid (AA). In terms of microflora, the addition of L, M, and LM made Firmicutes become the dominant bacterial phylum earlier, while Lactobacillus, Weissella, and Pediococcus had a higher abundance. According to the result of Pearson's correlation, there is a very significant negative correlation between pH value and Lactobacillus (P < 0.01) and a very significant positive correlation between pH value and Lactococcus, Enterobacter, Enterococcus, and Leuconostoc (P < 0.01), which may be inhibited by Lactobacillus under the decreased pH value. The results of the prediction of microbial genes indicated that the addition of M could enhance the carbohydrate metabolism and membrane transport metabolism, which may contribute to LA production by LAB metabolism. In general, L, M and LM all improved the fermentation quality and reduced the loss of nutrients to varying degrees, but considering the fermentation quality, the overall effects of M and LM were better than L. M and LM are recommended to be used as silage additives in the process of alfalfa silage in sandy grasslands to improve the quality.

The present study assessed the effects of oligosaccharide-chelated organic trace minerals (OTM) on the growth performance, digestive enzyme activity, blood parameters, slaughter performance, and meat quality indexes of mutton sheep. A total of 60 East Ujumuqin × small-tailed Han crossbred mutton sheep were assigned to two groups (10 duplicates per group) by body weight (26.12 ± 3.22 kg) according to a completely randomized design. Compared to the CON group, the results of the OTM group showed: (1) no significant changes in the initial body weight, final body weight, dry matter intake, average daily gain, and feed conversion ratio (p > 0.05); (2) the activities of trypsin, lipase, and amylase in the jejunum were significantly increased (p 0.05); (5) shear force of mutton was significantly lower (p 0.05). The content of crude protein was significantly higher (p 0.05). Among these, the essential amino acids, threonine, valine, leucine, lysine in essential amino acids and arginine were significantly increased (p

The fermentation substrate was supplemented with 0% guanidinoacetic acid (GAA) (control group, CON), 0.2% GAA (GAA02), 0.4% GAA (GAA04), 0.6% GAA (GAA06) and 0.8% GAA (GAA08) for 48 h of in vitro fermentation. Gas production was recorded at 2, 4, 6, 8, 12, 24, 36, and 48 h of fermentation. The gas was collected, and the proportions (%, v/v) of H2, CH4 and CO2 were determined. The rumen fermentation parameters, including pH, ammonia nitrogen (NH3-N), microbial protein (MCP) and volatile fatty acids (VFAs), were also determined. Furthermore, the bacterial community structure was analyzed through 16S rRNA high-throughput sequencing. The gene functions were predicted using PICRUSt1 according to the Kyoto Encyclopedia of Genes and Genomes (KEGG). The results showed that with the increase in GAA supplementation levels, the MCP and the concentration of rumen propionate were significantly increased, while the concentration of isovalerate was significantly decreased (p

Water scarcity is increasingly being recognized as a global concern. Sustainable Development Goal 6 (SDG-6) was established by the United Nations to address water resource governance within its sustainable development framework. This study employs the entropy weight method and a two-stage cyclical dynamic slacks-based measure (SBM) model to scientifically evaluate water resource utilization and governance across various regions in China. The findings reveal notable disparities in both the production and governance efficiency of water resources. Recognizing governance efficiency is crucial for promoting sustainable water resource utilization and socioeconomic development. The eastern, central, and western regions encounter unique challenges in attaining sustainability. The eastern region exhibits minimal potential for enhancing technical efficiency, necessitating interventions in management strategies and resource allocation. Conversely, the challenges in the central and western regions are more pronounced, demanding immediate implementation of new technologies and equipment. The data analysis in this study yields conclusions that offer targeted improvement recommendations to address disparities across China’s eastern, central, and western regions, and this is achieved by considering various developmental stages and regional contexts. These recommendations cover areas such as technical support, financial investment, and policy incentives, with the aim of enhancing the sustainable utilization of water resources in the country.

This study investigated the effects of ultrasound treatment on the quality of salted Culter alburnus fish. The results showed that with the increasing ultrasound power, the structural degradation of muscle fibers was intensified, and the conformation of myofibrillar protein was significantly changed. The high-power ultrasound treatment group (300 W) had relatively higher thiobarbiturate reactive substance content (0.37 mg malondialdehyde eq/kg) and peroxidation value (0.63 mmol/kg). A total of 66 volatile compounds were identified with obvious differences among groups. The 200 W ultrasound group exhibited fewer fishy substances (Hexanal, 1-Pentene-3-ol, and 1-Octane-3-ol). Compared with control group, ultrasound groups (200, 300 W) contained more umami taste-related amino peptides such as γ-Glu-Met, γ-Glu-Ala, and Asn-pro. In the ultrasound treatment group, L-isoleucine and L-methionine, which may be used as flavor precursors, were significantly down-regulated, while carbohydrates and its metabolites were up-regulated. Amino acid, carbohydrate, and FA (fatty acyls) metabolism products in salted fish were enriched by ultrasound treatment, and those products might ultimately be related to the taste and flavor of salted fish.

ABSTRACT This experiment was conducted to investigate the effects of lactic acid bacteria preparations on microbial diversity and community structure of calves. On days 1 and 7 of the trial period, feces were collected into sterile tubes and labeled (Day 1: control group D1DZ, experimental group D1SY, and Day 7: control group D7DZ, experimental group D7SY). Twenty Angus calves (150±10 kg) were selected and randomly divided into two groups of 10 calves each. The control group fed a basal diet. In addition to feeding the basal diet, the experimental group was given 15 mL lactobacillus preparation orally at 09:00 and 16:00 h every day. Calves were allowed free feeding and drinking water. All other feeding environments and management conditions remained consistent with the experiment lasting for seven days. At the end of the experiment, the fecal microflora of the calves was analyzed using 16S rRNA sequencing techniques. The 16S rRNA analysis data were processed using the Excel 2007 software and analyzed by the IBM SPSS statistical software (Statistical Analysis System, version 22). The Alpha diversity index analysis showed that the Chao and the Ace indices were significantly different after feeding supplemented with lactic acid bacteria. The PCA analysis showed that the fecal flora structure differed significantly after supplementation with the lactic acid bacteria preparation. Further analysis showed that the lactic acid bacteria increased Firmicutes, Patescibacteria, Rikenellaceae_RC9_gut_group, Clostridium_sensu_stricto_1, and prevotellaceae_UCG-003 in the feces. Therefore, we speculate that lactic acid bacteria preparations play an important role in animal production and are beneficial to the diversity of the fecal microflora of the calves.

BackgroundEstimating surgical case duration accurately is an important operating room efficiency metric. Current predictive techniques in spine surgery include less sophisticated approaches such as classical multivariable statistical models. Machine learning approaches have been used to predict outcomes such as length of stay and time returning to normal work, but have not been focused on case duration. ObjectiveThe primary objective of this 4-year, single-academic-center, retrospective study was to use an ensemble learning approach that may improve the accuracy of scheduled case duration for spine surgery. The primary outcome measure was case duration. MethodsWe compared machine learning models using surgical and patient features to our institutional method, which used historic averages and surgeon adjustments as needed. We implemented multivariable linear regression, random forest, bagging, and XGBoost (Extreme Gradient Boosting) and calculated the average R2, root-mean-square error (RMSE), explained variance, and mean absolute error (MAE) using k-fold cross-validation. We then used the SHAP (Shapley Additive Explanations) explainer model to determine feature importance. ResultsA total of 3189 patients who underwent spine surgery were included. The institution’s current method of predicting case times has a very poor coefficient of determination with actual times (R2=0.213). On k-fold cross-validation, the linear regression model had an explained variance score of 0.345, an R2 of 0.34, an RMSE of 162.84 minutes, and an MAE of 127.22 minutes. Among all models, the XGBoost regressor performed the best with an explained variance score of 0.778, an R2 of 0.770, an RMSE of 92.95 minutes, and an MAE of 44.31 minutes. Based on SHAP analysis of the XGBoost regression, body mass index, spinal fusions, surgical procedure, and number of spine levels involved were the features with the most impact on the model. ConclusionsUsing ensemble learning-based predictive models, specifically XGBoost regression, can improve the accuracy of the estimation of spine surgery times.

In the present study, we investigated the effects of cellulase and Lactiplantibacillus plantarum (L. plantarum) on the fermentation quality, microbial diversity, gene function prediction, and in vitro rumen fermentation parameters of the Caragana korshinskii (C. korshinskii) silage. The experimental treatment groups included control (CK), cellulase (C), L. plantarum (L), and cellulase + L. plantarum (CL). Triplicate silos were sampled after 14 and 56 days of ensiling. The microbial diversity of C. korshinskii silage at 56 days was analyzed using Illumina MiSeq sequencing technology, and the effects of C. korshinskii silage on rumen fermentation were evaluated by the in vitro artificial rumen gas production method. The results showed that the addition of cellulose and L. plantarum treatments reduced ammonia-nitrogen (NH3-N), dry matter (DM), neutral detergent fiber (NDF), and acid detergent fiber content during ensiling. Compared with the CK group, higher lactic acid content was observed in the silage of the C and L groups, but the CL group had a higher acetic acid (AA) content. Compared with the CK group, the relative abundance of Lactiplantibacillus significantly increased, but that of Enterococcus, Weissella, Enterobacter, and Pediococcus significantly decreased in all other groups except the C group (p < 0.05). The results of gene function prediction were as follows: pyruvate kinase, 1-phosphofructokinase, and fructokinase were lactate production enzymes, which suggested the association of the high AA content in the CL group to the high abundance of 6-phosphate glucose dehydrogenase. The abundance of H + -transporting ATPase and ATP-binding cassette (ABC) transporters in the L and CL groups was higher than that in the CK groups. Metabolism of terpenoids and polyketides, mismatch repair, DNA replication, nucleotide excision repair, and homologous recombination increased in the CL group compared with those in the CK group. Compared with the CK group, NH3-N and microbial protein content and the degradation rates of DM and NDF increased in the L and CL groups. To conclude, the L and CL groups had increased Lactiplantibacillus abundance, improved fermentation quality, and high DM degradation rates compared with the CK group.

Most coalbed methane reservoirs in China are of low porosity, low permeability and with extensive structural coals, which is difficult to be reformed, resulting in low single well gas production and recovery rates.It is the the key target for the development of coalbed methane industry to choose high efficiency reservoir and well reconstruction technology to effectively improve low efficiency wells.This paper systematically dissects the reasons for low production under the influence of "geological reservoir conditions, engineering construction reforms and drainage management control", summarizes the related technologies and application effects of the secondary reconstruction of coalbed methane wells, and provides suggestions for targeted reforms of different types of low-efficiency wells.The secodary reconstructable low-yield factors in coalbed methane wells include insufficient fractures propagation, fractures/pipe coal fines blockage, and limited pressure drop area.During secondary reconstruction, the distribution of coal structure, primary fracture morphology, reservoir permeability, variations of gas and water production, and applicability of drainage equipment need to be considered.The secondary reconstruction technology covers physical, chemical, microbial, and other methods.Secondary hydraulic fracturing, indirect fracturing, and anhydrous fracturing in physical methods, as well as acidification and permeability increasing technology and foam acidification in chemical methods are widely used.In secondary reconstruction, the reservoir adaptability of the reconstruction technology should be analyzed based on the geological conditions, primary reconstruction results, engineering and drainage status, and the purpose of secondary reconstruction should be determined based on the engineering drainage situation.In the secondary reconstruction, re-damage of the reservoir should be avoided to achieve effective transformation and improve the coalbed methane production from single well and well patterns.

ObjectivesThe objective of the study was to explore the added value of whole-exome sequencing (WES) in abnormal fetuses with detailed prenatal ultrasound and postnatal phenotype with normal karyotype and chromosomal microarray analysis (CMA).MethodsParents of fetuses with structural abnormalities by prenatal ultrasound who consented to provide fetal samples were prospectively recruited from January 2017 to December 2019. With aneuploidies or cases with copy number variations (CNVs) excluded, WES was performed for cases with normal karyotype and CMA results. Detailed prenatal ultrasound and postnatal imaging or pathology features were recommended for further interpretation of genetic variants.ResultsWES was performed for 94 eligible fetuses, DNA samples of which were extracted from 53 parent–fetus trios and 41 proband-only fetal tissues. A diagnostic genetic variant was identified in 37 (39.4%) of 94 fetuses, and 34 (64.2%) were detected in 53 trios, which was significantly greater than 3 (7.3%) in 41 proband-only cases (p < 0.001). In 34 trios with diagnostic genetic variants, 23 (67.6%) were de novo and 11 (32.4%) were inherited with two homozygous and nine heterozygous variants. Fourteen (14.9%) of 94 fetuses had a variant of uncertain significance (VUS). Among 94 cases, six affected pregnancies continued and 88 terminated, and 57 of 88 terminated cases underwent postmortem examinations. With accurate phenotypes demonstrated by prenatal ultrasound and postnatal autopsies, the clinical phenotypes were correlated in 33 (89.2%) of 37 cases with specific genotypes, with the highest matching ratio in skeletal diseases (20/33, 60.6%).ConclusionWES has added value in the genetic diagnosis of abnormal fetuses with normal karyotypes and CMA, particularly in skeletal diseases. Using WES in various anomalous fetuses can broaden the understanding of prenatal phenotypes and genetic variants.

Objective To analyze the relationship between nicotine dependence and depression among the elderly of Dai ethnic group in Jinggu county, Yunnan Province. Methods By using the method of multi-stage stratified random sampling, 1 409 rural Dai people aged 60 and over in Jinggu county, Yunnan Province were investigated by face-to-face questionnaire. Results The current smoking rate of rural Dai elderly in Jinggu county, Yunnan Province was 18.6% (37.1% for males and 0.8% for females). The smoking rate of males was significantly higher than that of females (P < 0.01). At present, the nicotine dependence rate of smokers was 32.8% (32.4% for males and 50.0% for females). There was no significant difference in nicotine dependence among Dai elderly people of different genders (P > 0.05), age groups (P > 0.05), education (P > 0.05), and annual family per capita income (P > 0.05). The prevalence rate of depression in the surveyed population was 52.0% (49.4% for males and 54.5% for females). Multivariate Logistic regression model analysis showed that patients with nicotine dependence were more likely to suffer from depression than those without nicotine dependence (OR=1.517, 95%C%: 1.0662.158). Conclusion Nicotine dependence is positively correlated with depression in the elderly of Dai ethnic group in Yunnan Province. Taking effective measures to reduce the nicotine dependence rate of smokers is helpful to improve the mental health level of the elderly. [ABSTRACT FROM AUTHOR]

Converting from conventional tillage to no-tillage influences the soil aggregate-size distribution and thus soil organic carbon (SOC) stabilization. However, the dynamics of soil aggregation and the straw-derived carbon (C) incorporation within aggregate fractions are not well understood. An experiment was established in 2004 to test the effects of two treatments, no-tillage with residue (NT) and conventional tillage without residue (CT), on the soil aggregate-size distribution and SOC stabilization in a continuous maize (Zea mays L.) cropping system located in the semiarid region of northern China. Soil samples were collected from the 0–10 cm layer in 2008, 2010 and 2015, and were separated into four aggregate-size classes (>2, 0.25–2, 0.053–0.25, and 2 and 0.25–2 mm) and associated SOC concentration compared with CT. Additionally, to compare straw-derived C incorporation within NT and CT aggregate fractions, 13C-labeled straw was incubated with intact NT and CT soils. After 90 days, the highest proportion of 13C-labeled straw-derived C was observed in the >2 mm fraction, and this proportion was lower in NT than that in CT soil. Overall, we conclude that long-term continuous NT increased the proportion of macroaggregates and the C concentration within macroaggregates, and the physical protection provided by NT is beneficial for soil C sequestration in the continuous maize cropping system in semiarid regions of northern China.

Abstract Background Whole-exome sequencing (WES) over the last few years has been increasingly employed for clinical diagnosis. However, one caveat with its use is that it inevitably fails to detect disease-causative variants that occur within noncoding RNA genes. Our experience in identifying pathogenic variants in the noncoding RNU4ATAC gene, in a Chinese family where two successive foetuses had been affected by severe microcephaly, is a case in point. These foetuses exhibited remarkably similar phenotypes in terms of their microcephaly and brain abnormalities; however, the paucity of other characteristic phenotypic features had made a precise diagnosis impossible. Given that no external causative factors had been reported/identified during the pregnancies, we sought a genetic cause for the phenotype in the proband, the second affected foetus. Results A search for chromosomal abnormalities and pathogenic copy number variants proved negative. WES was also negative. These initial failures prompted us to consider the potential role of RNU4ATAC, a noncoding gene implicated in microcephalic osteodysplastic primordial dwarfism type-1 (MOPD1), a severe autosomal recessive disease characterised by dwarfism, severe microcephaly and neurological abnormalities. Subsequent targeted sequencing of RNU4ATAC resulted in the identification of compound heterozygous variants, one being the most frequently reported MOPD1-causative mutation (51G>A), whereas the other was a novel 29T>A variant. Four distinct lines of evidence (allele frequency in normal populations, evolutionary conservation of the affected nucleotide, occurrence within a known mutational hotspot for MOPD1-causative variants and predicted effect on RNA secondary structure) allowed us to conclude that 29T>A is a new causative variant for MOPD1. Conclusions Our findings highlight the limitations of WES in failing to detect variants within noncoding RNA genes and provide support for a role for whole-genome sequencing as a first-tier genetic test in paediatric medicine. Additionally, the identification of a novel RNU4ATAC variant within the mutational hotspot for MOPD1-causative variants further strengthens the critical role of the 5′ stem-loop structure of U4atac in health and disease. Finally, this analysis enabled us to provide prenatal diagnosis and genetic counselling for the mother’s third pregnancy, the first report of its kind in the context of inherited RNU4ATAC variants.

Polyhydramnios is sometimes associated with genetic defects. However, establishing an accurate diagnosis and pinpointing the precise genetic cause of polyhydramnios in any given case represents a major challenge because it is known to occur in association with over 200 different conditions. Whole exome sequencing (WES) is now a routine part of the clinical workup, particularly with diseases characterized by atypical manifestations and significant genetic heterogeneity. Here we describe the identification, by means of WES, of novel compound heterozygous truncating variants in the LMOD3 gene [i.e., c.1412delA (p.Lys471Serfs*18) and c.1283dupC (p.Gly429Trpfs*35)] in a Chinese family with two successive fetuses affected with polyhydramnios, thereby potentiating the prenatal diagnosis of nemaline myopathy (NM) in the proband. LMOD3 encodes leiomodin-3, which is localized to the pointed ends of thin filaments and acts as a catalyst of actin nucleation in skeletal and cardiac muscle. This is the first study to describe the prenatal and postnatal manifestations of LMOD3-related NM in the Chinese population. Of all the currently reported NM-causing LMOD3 nonsense and frameshifting variants, c.1412delA generates the shortest truncation at the C-terminal end of the protein, underscoring the critical role of the WH2 domain in LMOD3 structure and function. Survey of the prenatal phenotypes of all known LMOD3-related severe NM cases served to identify fetal edema as a novel presenting feature that may provide an early clue to facilitate prenatal diagnosis of the disease.