Transcriptomic and metabolomic dissection of skeletal muscle of crossbred Chongming white goats with different meat production performance.

BACKGROUND: The transcriptome and metabolome dissection of the skeletal muscle of high- and low- growing individuals from a crossbred population of the indigenous Chongming white goat and the Boer goat were performed to discover the potential functional differentially expressed genes (DEGs) and differential expression metabolites (DEMs). RESULTS: A total of 2812 DEGs were detected in 6 groups at three time stages (3,6,12 Month) in skeletal muscle using the RNA-seq method. A DEGs set containing seven muscle function related genes (TNNT1, TNNC1, TNNI1, MYBPC2, MYL2, MHY7, and CSRP3) was discovered, and their expression tended to increase as goat muscle development progressed. Seven DEGs (TNNT1, FABP3, TPM3, DES, PPP1R27, RCAN1, LMOD2) in the skeletal muscle of goats in the fast-growing and slow-growing groups was verified their expression difference by reverse transcription-quantitative polymerase chain reaction. Further, through the Liquid chromatography-mass spectrometry (LC-MS) approach, a total of 183 DEMs in various groups of the muscle samples and these DEMs such as Queuine and Keto-PGF1α, which demonstrated different abundance between the goat fast-growing group and slow-growing group. Through weighted correlation network analysis (WGCNA), the study correlated the DEGs with the DEMs and identified 4 DEGs modules associated with 18 metabolites. CONCLUSION: This study benefits to dissection candidate genes and regulatory networks related to goat meat production performance, and the joint analysis of transcriptomic and metabolomic data provided insights into the study of goat muscle development.

Characterization and Comparative Analysis of Whole-Transcriptome Sequencing in High- and Low-Fecundity Chongming White Goat Ovaries during the Estrus Phase.

Reproductive performance is one of the most important economic traits in the goat industry. Increasing the number of goats is an effective measure to improve production efficiency and reduce production costs. Ovaries are important reproductive organs in female mammals that directly affect the estrous cycle and reproductive abilities. Understanding the complex transcription network of non-coding RNAs (lncRNAs, circRNAs, and miRNAs) and messenger RNA (mRNA) could lead to significant insights into the ovarian regulation of the reproductive processes of animals. However, the whole-transcriptome analysis of the non-coding RNAs and mRNA of the ovaries in Chongming white goats between high-fecundity (HP) and low-fecundity (LP) groups is limited. In this study, a whole-transcriptome sequencing approach was used to identify lncRNA, circRNA, miRNA, and mRNA expression in the ovaries of Chongming white goats during the estrus phase using RNA-Seq technology. More than 20,000 messenger RNAs (mRNAs), 10,000 long non-coding RNAs (lncRNAs), 3500 circular RNAs (circRNAs), and 1000 micro RNAs (miRNAs) were identified. A total of 1024 differential transcripts (724 mRNAs, 112 lncRNAs, 178 circRNAs, and 10 miRNAs) existing between the HP and the LP groups were revealed through a bioinformatics analysis. They were enriched in the prolactin signaling pathway, the Jak-STAT signaling pathway, and the GnRH signaling pathway, as well as various metabolic pathways. Differentially expressed mRNAs (such as LYPD6, VEGFA, NOS3, TNXB, and EPHA2) and miRNAs (such as miR-10a-5p) play key roles in the regulation of goat ovaries during the estrus phase. The enrichment of pathways related to reproduction, such as the Hippo, Hedgehog, PI3K-AKT, and MAPK signaling pathways, suggests that they might be involved in the prolificacy of goat ovaries. Overall, we identified several gene modules associated with goat fecundity and provided a basis for a molecular mechanism in the ovaries of Chongming white goats.

chi-miR-99b-3p Regulates the Proliferation of Goat Skeletal Muscle Satellite Cells In Vitro by Targeting Caspase-3 and NCOR1.

We previously found that chi-miR-99b-3p was highly expressed in the skeletal muscle of 7-month-old (rapid growth period) goats and speculated that it may be associated with muscle development. To further investigate the role of chi-miR-99b-3p in goats, we found that chi-miR-99b-3p acted as a myogenic miRNA in the regulation of skeletal muscle development. Dual-luciferase reporter assays, qRT-PCR, and Western blot results confirmed that Caspase-3 and nuclear receptor corepressor 1 were direct targets for chi-miR-99b-3p as their expression was inhibited by this miR. Cell proliferation and qRT-PCR assays showed that chi-miR-99b-3p promoted proliferation through relevant targets and intrinsic apoptosis-related genes in goat skeletal muscle satellite cells (SMSCs), whereas inhibition of chi-miR-99b-3p had the opposite effect. Furthermore, integrative transcriptomic analysis revealed that overexpression of chi-miR-99b-3p induced various differentially expressed (DE) genes mainly associated with the cell cycle, relaxin signaling pathway, DNA replication, and protein digestion and absorption. Notably, most of the cell-cycle-related genes were downregulated in SMSCs after miR-99b-3p upregulation, including the pro-apoptosis-related gene BCL2. In addition, 47 DE miRNAs (16 upregulated and 31 downregulated) were determined by Small RNA-sequencing in SMSCs after chi-miR-99b-3p overexpression. Based on the KEGG enrichment analysis, we found that these DE miRNAs were involved in the biological pathways associated with the DE genes. Our study demonstrated that chi-miR-99b-3p was an effective facilitator of goat SMSCs and provided new insights into the mechanisms by which miRNAs regulate skeletal muscle growth in goats.

Ages of weaning influence the gut microbiota diversity and function in Chongming white goats.

To study the effect of weaning age on the gut microbiota diversity in the lambs of Chongming white goats, fresh feces from the lambs weaned at 30, 45, and 60 days of age were collected 3 days after weaning at 33, 48, and 63 days of age, for microbial composition analysis by 16S rRNA sequencing. The serum concentrations of lipid metabolites were also investigated at the fecal collection dates. Serum and feces from the ewe-reared groups at 33, 48, and 63 days of age were used as controls. The alpha diversity increased significantly after weaning and with the aging of the lambs. Levels of Ruminococcaceae, Lachnospiraceae, and Ruminococcus varied significantly according to the weaning treatment in lambs (P < 0.05). Butyrate-producing gut bacteria such as Ruminococcaceae_UCG-010, Ruminococcaceae_UCG-013, Ruminococcaceae_UCG-014, Ruminococcaceae_UCG-005, Ruminococcaceae_UCG-002, Lachnospiraceae_AC2044_group, and Lachnospiraceae_NK4B4 were identified as significantly increased genera (P < 0.05) in the feces of weaned Chongming white lambs. Additionally, the abundance of fiber degradation-associated bacteria including Ruminococcaceae_UCG-005, Ruminococcus_1, and Ruminococcus_2 significantly increased with lamb weaning age (P < 0.05). Correlation analysis showed that Lachnospiraceae_AC2044_group, norank_f__Bacteroidales_S24-7_group, and Ruminococcaceae_UCG_005 were negatively correlated, and Lachnoclostridium was positively correlated with levels of cholesterol, while Blautia showed positive correlation with low-density lipoprotein cholesterol in serum samples from weaned lambs. This study helped to understand the maturing development of gut microbiota in Chongming white goats under weaning stress. KEY POINTS: • Effects of weaning age on the gut microbiota diversity in Chongming white goat lambs were studied. • Some butyrate-producing gut bacteria were significantly increased after weaned. • Correlations of gut microbiota and lipid metabolites were analyzed.

Factors Affecting Preventive Treatment Outcomes for Patients With Newly Diagnosed Chronic Migraine and Their Compliance With Treatment Recommendations in Chongqing Province, China: An Open-Label Prospective Study With Retrospective Baseline.

Objective: This study aims to investigate the factors affecting the efficacy of first oral prophylaxis in patients with chronic migraine (CM) and to assess patient compliance with their medication regimens. Method: To identify the therapeutic effect of prevention medication in 740 patients with newly diagnosed CM that did not receive any preventive treatments after 4 weeks in an open-label prospective study with retrospective baseline from January 2016 to January 2018, the factors that may affect the outcomes of preventive treatment were analyzed based on the demographic characteristics, migraine characteristics, family history of headache, and history of medication overuse. Moreover, the patients were followed up to evaluate their compliance with and the side effects of the medication at 4 weeks and at 12 weeks. Results: After 4 weeks of prophylaxis, 94.3% (n = 698) of the patients persisted with taking the medicine. The treatment was effective for 61.7% of CM patients (n = 431) and ineffective for 38.3% (n = 267). The results showed that the effectiveness of the preventive treatment was related to the number of headaches per month, and the effect was better for patients with headaches for 15-20 days/month than for those with headaches for 26-30 days/month (OR = 2.78, 95% CI: 1.26-5.75, P = 0.006). After 12 weeks of treatment, only 34.5% (n = 255) of the patients persisted with taking the medicine. The most common reason for non-compliance in CM patients is appointment difficulty in a headache clinic (31.8%). Conclusion: The effect of CM prophylaxis was related to the frequency of headache. Only 34.5% of the patients continued to take medicine after 12 weeks of treatment, suggesting that patient compliance needs to be enhanced in the prophylaxis of CM. For the Chinese headache society, the best way to increase patient compliance should be treatment at dedicated headache centers and timely visits to headache specialists.

Altered expression of miRNAs and mRNAs reveals the potential regulatory role of miRNAs in the developmental process of early weaned goats.

MicroRNAs (miRNAs) play pivotal roles in growth, development, and stress responses. However, the regulatory function of miRNAs in early weaned goats remains unclear. Deep sequencing comparison of mRNA and miRNA expression profiles showed that 18 miRNAs and 373 genes were differentially expressed in pre- and post-weaning Chongming white goats. Bioinformatics analysis indicated that these differentially expressed genes are involved in cellular processes, developmental processes, and growth in terms of biological process analysis. KEGG analysis showed that downregulated genes were enriched in salivary secretion, bile secretion, vascular smooth muscle contraction, and calcium signaling pathways. Additionally, a miRNA-mRNA co-expression network of the 18 dysregulated miRNAs and their 107 target mRNAs was constructed using a combination of Pearson's correlation analysis and prediction by miRanda software. Among the downregulated miRNAs, two (chi-miR-206 and chi-miR-133a/b) were muscle development-related and the others were cell proliferation associated. Further RT-qPCR analysis confirmed that downregulated miRNAs (chi-miR-99b-3p, chi-miR-224, and chi-miR-10b-5p) were highly expressed in muscle tissues (heart, spleen, or kidney) of the rapid growth period (7-month old) in Chongming white goats. The results of the present study suggested that weaning induced cell proliferation repression in post-weaning goats, providing new insight into the mechanism of muscle development of goats, although additional details remain to be elucidated in the future.

Testing of diagnosis criteria of tension-type headache: A multicenter clinical study.

Objective Tension-type headache is *These authors contributed equally to this work. usually manifested as head pain without associated symptoms, and the validation of diagnostic criteria presented are lacking and highly required in the International Classification of Headache Disorders. The aim of the present study was to explore the diagnosis criteria of tension-type headache in a multicenter-based sample from Chongqing, China. Methods Clinical characteristics and demographics were systematically and prospectively collected between March 2014 and December 2015 from 15 participating hospitals in Chongqing, using a semi-structured face-to-face interview. All patients were asked to complete a headache diary for at least 4 weeks. Results Out of 1832 patients with headache, 150 patients (97 female/53 male, 44.56 ± 11.9 years old) were diagnosed with tension-type headache based on the standard International Classification of Headache Disorders, 3rd edition beta version, and interestingly, 114 (76%) patients were diagnosed with tension-type headache based on the alternative criteria. One patient was excluded because only two of the four characteristics were fulfilled. Thirty-five (23.3%) patients did not meet the alternative criteria because of associated symptoms, including mild nausea (n = 6), photophobia (n = 1), and phonophobia (n = 28). All patients with TTH had mild or moderate headaches, 98.0% of patients suffered from non-pulsating headaches, 99.3% of patients said their headaches were not aggravated by routine physical activity, and 77.3% of patients had bilateral headache. Conclusions Non-pulsating headaches and headaches that are not aggravated by routine physical activity may represent core criteria for screening patients with tension-type headache. Nausea might not be an exclusion feature for diagnosis of TTH, but an important criterion for screening. Further studies are needed.

Toxic effects of nanomaterial-adsorbed cadmium on Daphnia magna.

Chemical immobilization technologies involving the use of chemical absorbents such as nanomaterials have been recommended for the remediation of Cd-contaminated water and soil. The impact of nanomaterials or nanomaterials coexisting with other contaminants on aquatic organisms has been reported, but information on the toxic effects of nanomaterial-adsorbed cadmium (Nano-Cd) on aquatic organisms is lacking. This study aimed to investigate the acute and sub-acute toxicity of Nano-Cd on Daphnia magna by using a method developed based on the standard Organisation for Economic Co-operation and Development (OECD) 202 guidelines. The toxicity of cadmium chloride (Cd2+), nano-manganese dioxide-cadmium (nMnO2-Cd), 20nm nano-hydroxyapatite-cadmium (nHAP20-Cd), and 40nm nano-hydroxyapatite-cadmium (nHAP40-Cd) to D. magna was in the following order: Cd2+> nMnO2-Cd > nHAP20-Cd > nHAP40-Cd. Further, nMnO2-Cd, nHAP20-Cd, and nHAP40-Cd showed acute toxicity to D. magna of level II grade according to the Commission of the European Communities and OECD standards. Exposure to low and medium, but not high, Nano-Cd concentrations increased the activities of peroxidase, superoxide dismutase, catalase, and anti-superoxide anion. Thus, Nano-Cd, particularly at high concentrations, could exert oxidative damage in D. magna. An increase in Cd2+ and Nano-Cd concentrations gradually increased the malondialdehyde content, indicating cell membrane damage caused by the production of excessive O2-. Thus, the use of nanomaterials after adsorption of Cd is associated with a potential risk to aquatic organisms.

Investigation of Mesoporous Graphitic Carbon Nitride as the Adsorbent to Remove Ni (II) Ions.

The mesoporous graphitic carbon nitride (mpg-C3N4/r, r was defined as the initial silica/dicyandiamide mass ratio) was successfully synthesized by heating the mixture of silica and dicyandiamide in a nitrogen atmosphere. The morphology and structure of mpg-C3N4/r were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Brunauer-Emmett-Teller surface area measurement (BET), X-ray powder diffraction (XRD), and Fourier Transform Infrared spectroscopy (FT-IR). The adsorption performances of Ni (II) ions by mpg-C3N4/r were investigated. With increasing of r value, the BET specific surface area of the synthesized mpg-C3N4/r increased; the highest specific surface area of mpg-C3N4/1.5 increased up to 169.3 m2/g. This work shows that mpg-C3N4/1.5 is a promising, high-efficiency adsorbent that can be used to purify the water of a low Ni (II) ions concentration. The maximum adsorption capacity of Ni(II) ions by mpg-C3N4/1.5 was 15.26 mg/g. The adsorption properties of Ni (II) ions by mpg-C3N4/r complied well with pseudo-second-order kinetics and Langmuir isotherm model.

Clinical observation of the time course of raised intracranial pressure after subarachnoid hemorrhage.

The time course of intracranial pressure (ICP) after subarachnoid hemorrhage (SAH) is not well known. This retrospective study was conducted to investigate the occurrence and the dynamic variation of raised ICP post-SAH. ICP was prospectively studied in 120 patients with SAH who were admitted to neurocritical care within 24 h of hemorrhage. Patients underwent continuous ICP monitoring for at least 7 days, unless they died. Clinical status on admission, radiographic tests, treatment details and neurological outcome on discharge were analyzed in relation to ICP. The highest daily mean ICP and the day when ICP reduced to normal levels were assessed. Of the 120 patients studied, 112 (93.3 %) encountered ICP elevation whilst in hospital. The daily mean ICP was higher in Hunt and Hess grades IV-V patients than grades I-III patients (P = 0.01). The elevated ICP remained at a higher level for the initial 3 days (grades I-III patients) or 4 days (grades IV-V patients), after which the pressure decreased towards normal levels. The in-patient mortality was significantly increased in the high ICP variability group (P = 0.001), which was divided by the cutoff point using receiver operating characteristic curve analysis. Raised ICP mainly occurs within 8 days post-SAH, especially the initial 3-4 days. Those highlight the need for earlier management of ICP after SAH.

Pilot study of amitriptyline in the prophylactic treatment of medication-overuse headache: a 1-year follow-up.

OBJECTIVE: This study aims to evaluate the long-term efficacy of low-dose amitriptyline combined with abrupt withdrawal in outpatients of medication-overuse headache (MOH) in an open-label design. METHODS: We evaluated the effectiveness of early introduction of low-dose amitriptyline combined with abrupt withdrawal in outpatients with MOH over a 1-year observational period. The primary outcome measures were the reduction in number of headache days and days with use of acute headache medication after 3 months and after 12 months. A number of secondary outcome measures, as well as safety and tolerability, were assessed. The responders were defined as patients with ≥50% reduction in headache frequency from baseline and being without medication overuse. RESULTS: Thirty-three patients completed the study. Significant reductions in headache frequency and medication consumption were observed at both months 3 and 12, compared to baseline (P < 0.05 for all). Neither the primary nor the secondary endpoints differed significantly between months 3 and 12. At the 12-month follow-up, 58% of the patients were considered as responders (N = 19); 73% remained cured of MOH (N = 24); 64% had reverted to episodic headaches (N = 21); 27% had relapsed into MOH (N = 9). CONCLUSION: Given these results, early introduction of low-dose amitriptyline combined with abrupt withdrawal could be considered as a choice for patients with MOH.