Temperature-Controllable Liquid Crystalline Medium for Stereochemical Elucidation of Organic Compounds via Residual Chemical Shift Anisotropies.

The configuration elucidation of organic molecules continues to pose significant challenges in studies involving stereochemistry. Nuclear magnetic resonance (NMR) techniques are powerful for obtaining such structural information. Anisotropic NMR techniques, such as measurement of residual dipolar couplings (RDCs) and residual chemical shift anisotropies (RCSAs), complementing isotropic NMR parameters, provide relative configuration information. RCSAs provide valuable structural information, especially for nonprotonated carbons, yet have been severely underutilized due to the lack of an easily operational alignment medium capable of rapid transition from anisotropic to isotropic environments, especially in aqueous conditions. In this study, an oligopeptide-based alignment media (FK)4 is presented for RCSA measurements. Temperature variation manipulates the assembly of (FK)4, yielding tunable anisotropic and isotropic phases without the requirement of any special devices or time-consuming correction procedures during data analysis. Decent observed ΔΔRCSA values from sp3 carbons benefit the utilization of RCSA measurements in the structural elucidation of organic molecules highly composed with sp3 carbons. Moreover, the (FK)4 alignment medium is applicable for both RDC and RCSA measurements in one sample, further advancing the configuration analysis of molecules of interest.

Knowledge of urinary incontinence among women 6 weeks to 1 year after delivery.

BACKGROUND: Urinary incontinence (UI) is common among postpartum women, but many do not seek medical help due to limited knowledge. Understanding the level of knowledge about UI in this population is essential for improving care-seeking behaviors and implementing targeted interventions. OBJECTIVE: The objective was to examine knowledge regarding UI among postpartum women with UI within 6 weeks to 1 year after delivery. METHODS: A cross-sectional study was conducted at obstetric clinic in two level-three grade A hospitals in Shenzhen, China, from January 2023 to June 2023. Women in their 6 weeks to 1 year after delivery with UI were asked to complete a questionnaire comprising three sections: (1) demographic variable; (2) International Consultation on Incontinence Questionnaire Short Form (ICIQ-UI SF); and (3) The Urinary Incontinence Quiz (UIQ). RESULTS: A total of 1228 women completed the questionnaire. Their mean UIQ score was 6.63 ± 3.51 (minimum = 2, maximum = 15), indicating the deficiency of UI knowledge among Chinese postpartum women. A total of 86.4% of participants experienced slight or moderate postpartum UI. The results of multivariate linear regression models for UIQ reveal significant independent associations between questionnaire scores and two variables: experience in pelvic floor muscle training (PFMT) (p < 0.001) and UI treatment in the past (p < 0.001). The overall model fit was R2 = 0.559 (p < 0.001). The regression coefficients for the experience in PFMT and UI treatment in the past were 2.301 and 4.916, respectively. However, no other discernible factors were identified to distinguish between those with and without adequate knowledge. CONCLUSIONS: Postpartum women with UI within 6 weeks to 1 year after delivery had poor knowledge of UI. Targeted educational interventions focusing on PFMT and early treatment for UI are essential.

A Systematic Review of Interventions for Demoralization in Patients with Chronic Diseases.

BACKGROUND: Demoralization, a significant mental health concern in patients with chronic diseases, can have a large impact on physical symptom burden and quality of life. The present review aimed to evaluate the effectiveness of interventions for demoralization among patients with chronic diseases. METHOD: PubMed, Scopus, Embase, and Web of Science were systematically searched. Research on providing interventions to patients with chronic diseases that included quantitative data on demoralization was then systematically reviewed. RESULTS: Fourteen studies were included, most of which considered demoralization as a secondary outcome. Interventions included evidence-based meaning-centered psychotherapy, dignity therapy, psilocybin-assisted psychotherapy, and others. Ten studies used randomized controlled designs. Six of these investigated evidence-based meaning-centered therapy, and four investigated dignity therapy, showing the best empirical support for these intervention types. Most studies showed significant impacts on demoralization in patients with chronic diseases. CONCLUSION: This systematic review provides insights into potential psychological interventions for reducing demoralization in patients with chronic diseases. Randomized controlled designs and adequately powered samples, with demoralization as the primary outcome, are needed to more clearly evaluate its effectiveness.

Identification of calcium chelating peptides from peanut protein hydrolysate and absorption activity of peptide-calcium complex.

BACKGROUND: Peanut peptides have good chelating ability with metal ions. However, there are few studies on the chelation mechanism of peanut peptides with calcium and absorption properties of peptide-calcium complex. RESULTS: Peptides with high calcium chelating rate were isolated and purified from peanut protein hydrolysate (PPH), and the chelation rate of component F21 was higher (81.4 ± 0.8%). Six peptides were identified from component F21 by liquid chromatography-tandem mass spectrometry, and the frequency of acidic amino acids and arginine in the amino acid sequence was higher in all six peptides. Peanut peptide-calcium complex (PPH21-Ca) was prepared by selecting component F21 (PPH21). Ultraviolet analysis indicated that the chelate reaction occurred between peanut peptide and calcium ions. Fourier transform infrared analysis showed that the chelating sites were carboxyl and amino groups on the amino acid residues of peptides. Scanning electron microscopy revealed that the surface of peanut peptide had a smooth block structure, but the surface of the complex had a granular morphology. Caco-2 cell model tests revealed that the bioavailability of PPH21-Ca was 58.4 ± 0.5%, which was significantly higher than that of inorganic calcium at 37.0 ± 0.4%. CONCLUSION: Peanut peptides can chelate calcium ions by carboxyl and amino groups, and the peptide-calcium complex had higher bioavailability. This study provides a theoretical basis for the development of new calcium supplement products that are absorbed easily. © 2024 Society of Chemical Industry.

Whole-genome sequencing revealed genetic diversity, structure and patterns of selection in Guizhou indigenous chickens.

BACKGROUND: The eight phenotypically distinguishable indigenous chicken breeds in Guizhou province of China are great resources for high-quality development of the poultry industry in China. However, their full value and potential have yet to be understood in depth. To illustrate the genetic diversity, the relationship and population structure, and the genetic variation patterns shaped by selection in Guizhou indigenous chickens, we performed a genome-wide analysis of 240 chickens from 8 phenotypically and geographically representative Guizhou chicken breeds and 60 chickens from 2 commercial chicken breeds (one broiler and one layer), together with 10 red jungle fowls (RJF) genomes available from previous studies. RESULTS: The results obtained in this present study showed that Guizhou chicken breed populations harbored higher genetic diversity as compared to commercial chicken breeds, however unequal polymorphisms were present within Guizhou indigenous chicken breeds. The results from the population structure analysis markedly reflected the breeding history and the geographical distribution of Guizhou indigenous chickens, whereas, some breeds with complex genetic structure were ungrouped into one cluster. In addition, we confirmed mutual introgression within Guizhou indigenous chicken breeds and from commercial chicken breeds. Furthermore, selective sweep analysis revealed candidate genes which were associated with specific and common phenotypic characteristics evolved rapidly after domestication of Guizhou local chicken breeds and economic traits such as egg production performance, growth performance, and body size. CONCLUSION: Taken together, the results obtained from the comprehensive analysis of the genetic diversity, genetic relationships and population structures in this study showed that Guizhou indigenous chicken breeds harbor great potential for commercial utilization, however effective conservation measures are currently needed. Additionally, the present study drew a genome-wide selection signature draft for eight Guizhou indigenous chicken breeds and two commercial breeds, as well as established a resource that can be exploited in chicken breeding programs to manipulate the genes associated with desired phenotypes. Therefore, this study will provide an essential genetic basis for further research, conservation, and breeding of Guizhou indigenous chickens.

Pelvic organ prolapse after delivery: effects on sexual function, quality of life, and psychological health.

BACKGROUND: Living with pelvic organ prolapse (POP) after delivery negatively impacted women's daily living and was detrimental to their psychological health. AIM: The purpose of the present study was to explore the effects of POP after delivery on women's sexual function, quality of life (QoL), and psychological health within the first year following delivery. METHODS: A cross-sectional study was conducted with 640 female participants at obstetric clinic in 2 hospitals in Shenzhen, China. OUTCOMES: All women completed following questionnaires: short form of the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) on sexual function, Pelvic Floor Distress Inventory-20 (PFDI-20) on well-being and QoL, Symptom Checklist-90-Revised (SCL-90-R) on psychological health, and International Consultation on Incontinence Questionnaire Short Form. RESULTS: Women with symptomatic POP (N = 250) had higher mean scores (P < .05) than those without symptoms (N = 390) in PISQ-12, PFDI-20, and SCL-90-R domains. In the symptomatic POP group (N = 250), the sexually dysfunctional group (N = 137) had significantly higher mean scores (P < .001) than the sexually functional group (N = 113) in PISQ-12, PFDI-20, and SCL-90-R domains. Women with POP duration ≥6 months (N = 132) had significantly higher mean scores (P < .001) than women with POP duration <6 months (N = 118) in PISQ-12, PFDI-20, and SCL-90-R domains. The multiple linear regression model of PISQ-12 showed that constipation history, regular pelvic floor muscle training, parity, and symptomatic POP were independently correlated with the questionnaire score (P < .05). CLINICAL IMPLICATIONS: The significantly pervasive nature of POP often leads to the neglect of a substantial percentage of sexual problems and related emotional suffering, emphasizing the critical importance of raising awareness about this issue among clinicians and the general public. STRENGTHS AND LIMITATIONS: The current study brought attention to significant aspects of postpartum POP; the findings suggest issues concerning POP occurrences and their impacts on sexual function, QoL, and psychological health. Primary constraints are linked to the utilization of self-report assessments. CONCLUSION: The current study reveals a robust correlation between POP in postpartum women and compromised sexual function, QoL, and psychological health.

Development and validation of a Decision-Making Ability Scale for postpartum urinary incontinence women engaging in pelvic floor physical therapy.

OBJECTIVE: This study aimed to develop and validate a Decision-Making Ability Scale (DMA-S) for postpartum urinary incontinence (PPUI) women engaging in pelvic floor physical therapy (PFPT). METHODS: Items were created in line with a review of the literature and exploratory qualitative study with 22 women. The items were submitted for expert opinion and a pilot implementation was made with 58 women with PPUI. Furthermore, the construct validity of the scale was tested with exploratory factor analysis (EFA) (n = 220) and confirmatory factor analysis (CFA) (n = 240). Internal consistency for the Chronbach's α and test-retest reliability for the intraclass correlation coefficient (ICC) were also investigated for the DMA-S in the study. RESULTS: The results of the EFA indicated a Kaiser-Meyer-Olkin value of 0.85 and Bartlett's test of sphericity showed a χ2 value of 8352.101, p < 0.001. After removing one item with factor loading values below 0.50, the resulting factor structure accounted for 83.38% of the total variance. The fit indices of the scale model tested in the CFA were determined as χ2 /df = 1.08 < 3, root mean square error of approximation = 0.018 < 0.08, comparative fit index = 0.996 > 0.90, Tucker-Lewis index = 0.995 > 0.90, goodness-of-fit index (GFI) = 0.933 > 0.90, adjusted GFI = 0.916 > 0.90, and incremental fit index = 0.996 > 0.90. The Cronbach's α values were 0.95-0.97 for the subdimensions of the scale and 0.93 for the total scale. Data also showed a good test-retest stability (ICC = 0.984). CONCLUSION: The DMA-S is a reliable and valid tool for assessing the decision-making ability for PPUI women engaging in PFPT.

Bioinformatics-based analysis of the roles of sex hormone receptors in endometriosis development.

Endometriosis is a hormone-dependent disease in women of reproductive age and seriously affects women's health. To analyze the involvement of sex hormone receptors in endometriosis development, we performed bioinformatics analysis using four datasets derived from the Gene Expression Omnibus (GEO) database, which may help us understand the mechanisms by which the sex hormones act in vivo in endometriosis patients. The enrichment analysis and protein-protein interaction (PPI) analysis of the differentially expressed genes (DEGs) revealed that there are different key genes and pathways involved in eutopic endometrium aberrations of endometriosis patients and endometriotic lesions, and sex hormone receptors, including androgen receptor (AR), progesterone receptor (PGR) and estrogen receptor 1 (ESR1), may play important roles in endometriosis development. Androgen receptor (AR), as the hub gene of endometrial aberrations in endometriotic patients, showed positive expression in the main cell types for endometriosis development, and its decreased expression in the endometrium of endometriotic patients was also confirmed by immunohistochemistry (IHC). The nomogram model established based on it displayed good predictive value.

Study on the role of heat shock protein 90 (HSP90) gene in chicken preadipocytes proliferation and differentiation.

In this study, we examined the effects of Heat Shock Protein 90 (HSP90) on adipocyte proliferation and differentiation in chickens. To achieve this, we constructed RNA interference (RNAi) vectors to target HSP90 and transfected the vectors into primary adipocytes. After transfection, oil red O staining was performed to determine the status of triglyceride accumulation in the cells, whereas the CCK-8 cell kit and 5-Ethynyl-2'-Deoxyuridine (EdU) assays were used to determine cell proliferation. Thereafter, the mRNA and protein expression levels of PPARγ, FAS, SREBP-1c, and HSP90 were determined, and the results showed that after the interference of HSP90, the mRNA and protein expression levels of HSP90 in the chicken adipocytes decreased significantly compared to the control and blank groups (p < 0.05). The decreased mRNA and protein expression of PPARγ, FAS, and SREBP-1c was related to adipocyte differentiation (p < 0.05). However, HSP90 interference had no effect on adipocyte proliferation (p > 0.05). Taken together, the results of this study showed that HSP90 influenced the expression of PPARγ and adipose-differentiation-related genes, thereby regulating triglyceride accumulation and adipocyte differentiation in chickens.

BIBR1532 Affects Endometrial Cell Proliferation, Migration, and Invasion in Endometriosis via Telomerase Inhibition and MAPK Signaling.

OBJECTIVES: The effect of telomerase inhibitor BIBR1532 on endometriotic cells was investigated to explore the inhibitory effect of targeting telomerase on endometriosis. DESIGN: In vitro primary cell culture study. Participants/Materials: Primary endometrial cells derived from eutopic and ectopic endometrium in patients with endometriosis. SETTING: The study was conducted in the university hospital. METHODS: Paired eutopic and ectopic endometrial cells were collected from 6 patients from January 2018 to July 2021. A TRAP assay was performed to detect the telomerase activity of the cells. MTT, cell cycle, apoptosis, migration, and invasion assays were performed to study the inhibitory effect of BIBR1532. Enrichment analysis was performed to identify the key pathways involved in endometriosis progression and telomerase action. Then, Western blotting was used to investigate the expression of related proteins. RESULTS: BIBR1532 treatment significantly inhibited the growth of eutopic and ectopic endometrial cells, with apoptosis and cell cycle signaling involved. Migration and invasion, important characteristics for the establishment of ectopic lesions, were also inhibited by BIBR1532. The MAPK signaling cascade, related to telomerase and endometriosis, was decreased in eutopic and ectopic endometrial stromal cells with the treatment of BIBR1532. LIMITATIONS: The severe side effects of telomerase inhibitors might be the main obstacle to clinical application, so it is necessary to find better drug delivery methods in vivo. CONCLUSIONS: The telomerase inhibitor BIBR1532 affects endometrial cell proliferation, migration, and invasion in endometriosis.

Effects of Dietary Ferulic Acid on Intestinal Health and Ileal Microbiota of Tianfu Broilers Challenged with Lipopolysaccharide.

Lipopolysaccharide (LPS) has been considered the primary agent to establish animal models of inflammation, immunological stress, and organ injury. Previous studies have demonstrated that LPS impaired gastrointestinal development and disrupted intestinal microbial composition and metabolism. Ferulic acid (FA) isolated from multiple plants exhibits multiple biological activities. This study investigated whether FA ameliorated intestinal function and microflora in LPS-challenged Tianfu broilers. The results showed that LPS challenge impaired intestinal function, as evidenced by decreased antioxidant functions (p < 0.05), disrupted morphological structure (p < 0.05), and increased intestinal permeability (p < 0.05); however, these adverse effects were improved by FA supplementation. Additionally, FA supplementation preserved sIgA levels (p < 0.05), increased mRNA expression levels of CLDN and ZO-1 (p < 0.05), and enhanced epithelial proliferation (p < 0.05) in the ileal mucosa in LPS-challenged chickens. Moreover, FA supplementation rectified the ileal microflora disturbances in the LPS-challenged broilers. The results demonstrate that dietary FA supplementation decreased LPS-induced intestinal damage by enhancing antioxidant capacity and maintaining intestinal integrity. Furthermore, FA supplementation protects intestinal tight junctions (TJs), elevates secretory immunoglobulin A (sIgA) levels, and modulates ileal microflora composition in LPS-challenged broilers.

OsmiR396/growth regulating factor modulate rice grain size through direct regulation of embryo-specific miR408.

microRNAs (miRNAs) are promising targets for crop improvement of complex agricultural traits. Coordinated activity between/among different miRNAs may fine-tune specific developmental processes in diverse organisms. Grain size is a main factor determining rice (Oryza sativa L.) crop yield, but the network of miRNAs influencing this trait remains uncharacterized. Here we show that sequestering OsmiR396 through target mimicry (MIM396) can substantially increase grain size in several japonica and indica rice subspecies and in plants with excessive tillers and a high panicle density. Thus, OsmiR396 has a major role related to the regulation of rice grain size. The grain shape of Growth Regulating Factor8 (OsGRF8)-overexpressing transgenic plants was most similar to that of MIM396 plants, suggesting OsGRF8 is a major mediator of OsmiR396 in grain size regulation. A miRNA microarray analysis revealed changes to the expression of many miRNAs, including OsmiR408, in the MIM396 plants. Analyses of gene expression patterns and functions indicated OsmiR408 is an embryo-specific miRNA that positively regulates grain size. Silencing OsmiR408 expression (miR408KO) using CRISPR technology resulted in small grains. Moreover, we revealed the direct regulatory effects of OsGRF8 on OsMIR408 expression. A genetic analysis further showed that the large-grain phenotype of MIM396 plants could be complemented by miR408KO. Also, several hormone signaling pathways might be involved in the OsmiR396/GRF-meditated grain size regulation. Our findings suggest that genetic regulatory networks comprising various miRNAs, such as OsmiR396 and OsmiR408, may be crucial for controlling rice grain size. Furthermore, the OsmiR396/GRF module may be important for breeding new high-yielding rice varieties.

High sensitivity fiber cladding SPR strain sensor based on V-groove structure.

How to couple the light in the fiber core to the cladding is an urgent issue that need to be done for the fabrication of the fiber-cladding SPR sensor, and there is no report about the fiber SPR strain sensor. Hereby, we propose and demonstrate a high sensitivity fiber cladding SPR strain sensor based on V-groove structure. By CO2 laser, the V-groove is fabricated on the single-mode fiber, and the light in the fiber core is effectively coupled to the cladding. The cladding 2cm behind the V-groove is coated with sensing gold film, and a multimode fiber is spliced with the sensing probe to construct the novel fiber cladding SPR sensor. On the basis of the investigation of the effects of different V-groove depth, number and period on the performance of fiber SPR refractive index sensor, a high sensitivity strain SPR sensor is designed and fabricated by employing the characteristic that the V-groove will deform with strain. The testing results indicate that the average refractive index sensitivity of the sensor is 2896.4nm/RIU, and the strain wavelength sensitivity is 25.92pm/µÎµ which is much higher than that of the fiber interference and grating strain sensors, and the strain light intensity sensitivity is -4.4×10-4 a.u./µÎµ. The proposed fiber cladding SPR strain sensor has the advantages of simple structure and convenient manufacture, and can be used for working in a narrow space.

MUSTN1 is an indispensable factor in the proliferation, differentiation and apoptosis of skeletal muscle satellite cells in chicken.

The yield and quality of the skeletal muscle are important economic traits in livestock and poultry production. The musculoskeletal embryonic nuclear protein 1 (MUSTN1) gene has been shown to be associated with embryonic development, postnatal growth, bone and skeletal muscle regeneration; however, its function in the skeletal muscle development of chicken remains unclear. Therefore, in this study, we observed that the expression level of MUSTN1 increased in conjunction with the proliferation of chicken skeletal muscle satellite cells (SMSCs). Knockdown of MUSTN1 in SMSCs downregulated the expression of cell proliferation genes as Pax7, CDK-2 and differentiation-relate genes including MyoD, MyoG, MyHC and MyH1B, whereas it upregulates the expression of cell apoptosis gene (Caspase-3) (P < 0.05). However, the combined analysis of CCK-8 and EdU showed that the cell vitality and EdU-positive cells of the si-MUSTN1 transfected group were significantly lower than those of the negative siRNA group (P < 0.05). In addition, the knockdown of MUSTN1 significantly increased the cell population in the G0/G1 phase and significantly decreased the cell population in the G2/M phase (P < 0.05), whereas the overexpression of MUSTN1 showed opposite effect. Taken together, our findings indicates that MUSTN1 is an important molecular factor that is responsible for regulating muscle growth and development in chickens, particularly, proliferation and differentiation of SMSCs.

S-type fiber surface plasmon resonance strain sensor.

The fiber surface plasmon resonance (SPR) sensor is widely used in high-sensitivity refractive index detection, and there are few reports on SPR sensors used for strain measurement. In this paper, we propose and demonstrate an S-type fiber strain sensor based on SPR. The simulation and testing results indicate that the smaller the vertical axis offset of S-type fiber is, the larger the incidence angle of SPR is, and the closer the working band of SPR is to the shorter wavelength direction. By electrofusion, we fabricated an S-type structure on the single-mode fiber, and by the S-type structure, the high-order cladding mode was excited. A 50 nm gold film was coated on the surface of the fiber cladding behind the S-type structure. The evanescent field of the cladding mode contacted the gold film to produce SPR, and the strain can change the vertical axis offset of the S-type fiber and further change the incidence angle of SPR; hence an S-type fiber strain sensor based on SPR was realized. When the refractive index of the ambient medium is 1.345 RIU, the vertical axis offset and length of the S-shaped structure are 87 and 501 µm, respectively, the resonance wavelength of the fiber SPR strain sensor changes from 648.06 to 631.77 nm with a strain detection range of 0-1200µÎµ, and its sensitivity is -14.38pm/µÎµ. The proposed sensor provides a new solution for the strain measurement of the fiber SPR sensor, which is expected to be used in the fields of engineering, health monitoring, and early warning.

Single-mode fiber curvature sensor based on SPR.

A fiber surface plasmon resonance (SPR) sensor is widely used in high-sensitivity refractive index measurement, but there is less research on curvature measurement. In this paper, a single-mode fiber curvature sensor based on SPR is designed and fabricated. By employing bending, the transmitted light in the fiber core leaks into the cladding. A 50 nm gold film is coated outside the cladding, and the evanescent field of the cladding after bending contacts the gold film to cause SPR. When the curvature changes, the coupled cladding mode and intensity are different; that is, the SPR incident angle and evanescent field intensity are different, so as to realize the dual parameters of SPR resonance wavelength and depth of the resonance valley changing with curvature. By experiments, the influence of different cutoff wavelengths of single-mode fiber on the performance of the sensor is studied. The testing results indicate that with the decrease in cutoff wavelength of the single-mode fiber, the valley depth sensitivity of the sensor increases, and the half height width (FWHM) decreases. When the cutoff wavelength of the single-mode fiber is 630 nm, the valley depth sensitivity of the sensor is 0.0088a.u/m-1, the wavelength sensitivity is 0.26nm/m-1, and the average FWHM is only 21 nm. The proposed single-mode fiber curvature sensor based on SPR has a narrow FWHM and an opening threshold. It can also realize no opening threshold by introducing a coreless fiber, which provides a new solution, to the best of our knowledge, for the diversified detection of fiber SPR sensors.

Comprehensive analysis of differently expression mRNA and non-coding RNAs, and their regulatory mechanisms on relationship in thiram-induced tibial dyschondroplasia in chicken.

Thiram pollution is one of the main causes of tibial dyschondroplasia (TD) induced by feed sources. Several studies have speculated that miRNA, circRNA and lncRNA may have significant impact on the development of TD, however, the specific mRNAs and noncoding RNAs and their respective regulatory mechanisms and functions in the development of TD have not been explored. Therefore, in this present study, we screened the differentially expressed mRNA, miRNA, circRNA and lncRNA by whole-transcriptome sequencing (RNA-seq) and differentially expressed genes (DEGs) enrichment, as well as constructed the interaction network among the mRNA-miRNA, mRNA-lncRNA and mRNA-miRNA-circRNA. The sequencing results were verified by fluorescence real-time quantitative PCR (RT-qPCR). The results obtained in this study, revealed that the cells were atrophied and disordered in the TD group, and the expression of BMP6, TGF-ß and VEGF were significantly reduced. A total of 141 mRNAs, 10 miRNAs, 23 lncRNAs and 35 circRNAs of DEGs were obtained (p<0.05) Theses DEGs were enriched in the adhere junction and insulin signaling pathways. In addition, the mRNA-miRNA-circRNA network suggested that several pivotal ceRNA showed a regulatory relationship between the transcripts with miRNA, circRNA or lncRNA. Taken together, the results in the present study, represent an insight for further functional research on the ceRNA regulatory mechanism of TD in broilers.

[Analysis of child with pyruvate carboxylase deficiency type A due to compound heterozygous variants of the PC gene].

OBJECTIVE: To analyze the clinical features and genetic basis for a child with pyruvate carboxylase deficiency type A (PCD-A). METHODS: Clinical data of the child was retrospectively analyzed. The child and his parents were subjected to trio-whole exome sequencing, and candidate variants were verified by bioinformatics analysis. RESULTS: The child was admitted due to fever with vomiting and disturbance of consciousness. His clinical manifestations included severe decompensated acidosis, hypotension and intractable shock. Cranial MRI showed abnormal signal in the brain, and chest X-ray revealed acute pulmonary edema. DNA sequencing revealed that he has harbored compound heterozygous variants of the PC gene, namely c.182T>C (p.I61T) and c.2581G>A (p.V861M), which were respectively inherited from his father and mother. Neither variant was retrievable in the ClinVar and HGMD databases. Through prediction of protein structure, both variants may affect the functional stability of the protein product. CONCLUSION: The compound heterozygous variants of the PC gene probably underlay the PCD-A in this child. Combined with the clinical features, the child was ultimately diagnosed as PCD-A. Above finding has enriched the spectrum of PC gene variation underlying PCD-A.

Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens.

BACKGROUND: miRNAs regulate circadian patterns by modulating the biological clocks of animals. In our previous study, we found that the clock gene exhibited a cosine expression pattern in the fallopian tube of chicken uterus. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in the chicken uterus and, if so, how they regulate egg-laying rhythms is unclear. In this study, we selected 18 layer hens with similar ovipositional rhythmicity (each of three birds were sacrificed for study per 4 h throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens. RESULTS: We identified six circadian miRNAs that are mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of the experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in the chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in the chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in the chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. The expression of miR-449c-5p showed an opposite trend to that of ATP2B4 within a 24 h cycle in the chicken uterus; it inhibited mRNA and protein expression of ATP2B4 in the uterine tubular gland cells. In addition, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. We found similar results after ATP2B4 knockdown by miR-449c-5p. Taken together, these results indicate that ATP2B4 promotes uterine Ca2+ trans-epithelial transport. CONCLUSIONS: Clock-controlled miR-449c-5p regulates Ca2+ transport in the chicken uterus by targeting ATP2B4 during eggshell calcification.

A functional polymorphism of inhibin alpha subunit at miR-181b-1-3p-binding site regulates proliferation and apoptosis of chicken ovarian granular cells.

INHA, the gene encoding the inhibin alpha subunit, was involved in folliculogenesis in mammals, but no study was reported for its working pathway in birds. Here we hypothesize that gene polymorphism in INHA 3'UTR might influence miRNAs binding efficiency and further affect the function of this gene. Thus, we investigated the association between the 3'UTR single-nucleotide polymorphisms (SNPs) in INHA and the laying performance in chickens and further explore their possible molecular cascades in granulosa cells (GC). Five SNPs were detected in Tianfu green-shell layers and g. 22,178,975 G > A was significantly associated with total egg numbers at the age of 300 days (EN, n = 286). Birds carrying the AA genotype laid more EN than those with GG (P < 0.05). The allele transition from G to A in the 3'UTR of INHA gene destroyed a binding site which was targeted by miR-181b-1-3p. The expression abundances of INHA mRNA increased firstly and then decreased with follicle growing, and reached the top in the sixth largest pre-ovulation follicle, whereas miR-181b-1-3p levels in chicken pre-hierarchical follicles had the contrary tendency. Further studies indicated that high levels of miR-181b-1-3p increased apoptosis and reduced GC proliferation while miR-181b-1-3p inhibitors decreased apoptosis and promoted GC proliferation. Additionally, depression of INHA increased apoptosis and reduced GC proliferation via a caspase-3-dependent mitochondrial pathway. Generally, the mutation in INHA 3'UTR was tightly correlated with egg production in chickens, and blocked a binding site of miR-181b-1-3p. miR-181b-1-3p inhibited GC proliferation and promoted apoptosis by targeting INHA.