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ABSTRACT: Within the poultry industry, hens' reproductive performance is of great economic significance. The development and growth of follicles is a key aspect of hen egg production, and ovarian follicle growth and development are closely associated with granulosa cells (GCs) proliferation and the synthesis of steroid hormones. It has been confirmed by numerous studies that microRNAs (miRNAs) play important roles in the steroid hormone synthesis and proliferation of GCs. In this study, we examined the main miRNAs influencing hens' ability to reproduce, identified the miR-223 that is mainly expressed in atretic follicles based on sequencing, and investigated its role in GCs. Then, we used miR-223 mimic and inhibitor to knockdown or overexpress miR-223 expression. The result showed that miR-223 significantly inhibits both the steroid hormone synthesis and the proliferation of GCs. Subsequently, the results of the dual luciferase reporter experiment and bioinformatics prediction demonstrated that cysteine rich transmembrane BMP regulator 1 (CRIM1) was a downstream target gene of miR-223, and overexpression of miR-223 prevented CRIM1 expression. The function of CRIM1 was further investigated, and we observed a significant reduction in the synthesis of steroid hormones and the proliferation of GCs after transfection with CRIM1 siRNA. The opposite function of miR-223 was observed for CRIM1 in our study. Additionally, we demonstrated the involvement of the miR-223/CRIM1 axis in GCs through modulation of the AKT signaling pathway. Our data demonstrate the pivotal role of the miR-223 in the proliferation and steroid hormone synthesis of chicken GCs, which helps to explain how non-coding RNA (ncRNA) affects chicken reproductive function.

ABSTRACT: Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs that have been implicated in mediating granulosa cell (GC) proliferation and apoptosis. CircRAB11A was found to have a significantly higher expression in normal follicles compared to atrophic follicles. In this study, we determined that the knockdown of circRAB11A resulted in the inhibition of proliferation and promotion of apoptosis in GCs of chicken. Moreover, circRAB11A was found to act as a sponge for miR-24-5p, both member RAS oncogene family (RAB11A) and epidermal growth factor receptor (EGFR) were revealed to be targets of miR-24-5p through a dual-luciferase reporter assay. RAB11A or EGFR promoted proliferation and suppressed apoptosis in GCs through the phosphatidylinositol-kinase (PI3K)/AKT or extracellular signal-regulated kinase (ERK)1/2 pathway. These findings suggest that circRAB11A may function as a competing endogenous RNA (ceRNA) by targeting the miR-24-5p/RAB11A and miR-24-5p/EGFR axes and activating the ERK1/2 and PI3K/AKT pathways, offering a potential avenue for exploring the mechanism of follicle development.

As a zoonotic parasite, Cryptosporidium spp. could cause severe diarrhea mainly in calves and children globally. Monitoring and prevention of Cryptosporidium spp.’s prevalence is of great significance in both economy and public health aspects. In this study, specific primers and probes were designed within the conserved region of 18S rRNA gene for Cryptosporidium spp. and recombinase polymerase amplification assays based on the fluorescence monitoring (real-time RPA) as well as combined with a lateral flow strip (LFS RPA) were developed. Both of the two RPA assays allowed the exponential amplification of the target fragment within 20 min. After incubation on a metal bath at 42 °C, the LFS RPA results were displayed on the lateral flow strip within 5 min while real-time RPA allowed the real-time observation of the results in Genie III at 39 °C. The RPA assays showed high specificity for Cryptosporidium spp. without any cross-reaction with other tested pathogens causing diarrhea in cattle. With the recombinant plasmid DNA containing the 18S rRNA gene of Cryptosporidium spp. serving as a template, the limit of detection for real-time RPA and LFS RPA assays were 14.6 and 12.7 copies/reaction, respectively. Moreover, the RPA assays were validated by testing diarrheic cattle fecal samples and compared with a real-time PCR. The positive ratio of Cryptosporidium spp. was 24.04 % (44/183) and 26.23 % (48/183) in both RPA assays and real-time PCR assay, respectively, and the kappa coefficient value was 0.942. The diagnostic specificity and diagnostic sensitivity of both RPA assays were 100 % and 91.67 %, respectively. Forty-one of 48 positive samples were successfully sequenced and four Cryptosporidium species were detected, including C. parvum (n = 20), C. andersoni (n = 17), C. bovis (n = 3) and C. ryanae (n = 1). The developed RPA assays are easy to operate and faster to obtain the detection results, and they are suiting for the point-of-care detection and facilitating the prevention and control of Cryptosporidium spp. infections.

Duck is often used in meat fraud as a substitute for more expensive meats. Rapid detection of duck ingredient in meat products is of great significance for combating meat fraud and safeguarding the interests of consumers. Therefore, we aim to develop duck-specific recombinase polymerase amplification (RPA)-based assays for the rapid detection of duck ingredient in animal-derived foods. Using Cytb gene as target, the real-time RPA and RPA combined with lateral flow strips (LFS RPA) were developed successfully for the rapid detection of ducks in 20 min at 39 °C and 40 °C, respectively. The assays did not show cross-reactions with 6 other livestock and poultry. The developed RPA assays could detect 10 pg duck genomic DNA per reaction and 0.1 % (w/w) duck ingredient in duck and mutton mixed powder within 30 min, including a rapid nucleic acid extraction. Furthermore, duck ingredient could be detected in 30 different actual foods including heat-processed meats and blood products. Therefore, duck-specific real-time RPA and LFS RPA assays were successfully developed with good specificity and sensitivity, which could enable rapid detection of duck ingredient in the field and provide technical support for combating the meat fraud.

Abstract Background Radiation-induced insufficiency fractures (IF) is frequently occult without fracture line, which may be mistaken as metastasis. Quantitative apparent diffusion coefficient (ADC) shows potential value for characterization of benign and malignant bone marrow diseases. The purpose of this study was to develop a nomogram based on multi-parametric ADCs in the differntiation of occult IF from bone metastasis after radiotherapy (RT) for cervical cancer. Methods This study included forty-seven patients with cervical cancer that showed emerging new bone lesions in RT field during the follow-up. Multi-parametric quantitative ADC values were measured for each lesion by manually setting region of interests (ROIs) on ADC maps, and the ROIs were copied to adjacent normal muscle and bone marrow. Six parameters were calculated, including ADCmean, ADCmin, ADCmax, ADCstd, ADCmean ratio (lesion/normal bone) and ADCmean ratio (lesion/muscle). For univariate analysis, receiver operating characteristic curve (ROC) analysis was performed to assess the performance. For combined diagnosis, a nomogram model was developed by using a multivariate logistic regression analysis. Results A total of 75 bone lesions were identified, including 48 occult IFs and 27 bone metastases. There were significant differences in the six ADC parameters between occult IFs and bone metastases (p

Abstract Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has become a public health emergency of international concern. SARS-CoV-2 RNA detection is the diagnostic criterion for coronavirus disease 2019 (COVID-19). Nevertheless, RNA detection has many limitations, such as being time-consuming and cost-prohibitive, and it must be performed in specialized laboratories. Virus antibody detection is a routine method for screening for multiple viruses, but data about SARS-CoV-2 antibody detection are limited. Method Throat swabs and blood were collected from 67 suspected SARS-CoV-2 infection patients at the Affiliated Hospital of Zunyi Medical University and Zunyi Fourth People’s Hospital isolated observation departments. Throat swab samples were subjected to SARS-CoV-2 RNA detection by real-time PCR. Blood was used subjected to SARS-CoV-2 IgG/IgM detection by an enzyme-linked immunosorbent assay (ELISA) and gold immunochromatography assay (GICA). Blood underwent C-reactive protein detection by immunoturbidimetry, and white blood cells, neutrophil percentages and lymphocyte percentages were counted and calculated, respectively. Clinical symptoms, age and lifestyle habits (smoking and drinking) in all patients were recorded. Data were analysed using SPSS version 19. The results were confirmed by T and χ2 tests; correlations with detection results were analysed by kappa coefficients. Odds ratio (OR) and corrected OR values were analysed by logistic regression. P

Objective To establish a real-time recombinase-aid amplification (RAA) method for rapid detection of Pseudomonas aeruginosa. Methods Specific primers and exo probes based on ecfX gene of P. aeruginosa were designed in this study, and the validity of the method was evaluated by sensitivity, specificity and suspected strains detection. Results Real-time RAA was performed successfully at 39℃ for 20 min. Only the P. aeruginosa strains but not other bacteria were amplified, showing the good specificity. The limit of detection was 3.0×103 fg genomic DNA per reaction, and 1.0×103 CFU P. aeruginosa pure culture per reaction. The developed real-time RAA was further evaluated on 36 suspected of P. aeruginosa, which were identified successfully to be P. aeruginosa.The detection result were the same with those of a real-time PCR assay and the VITEK 2 Compact. Conclusion The developed real-time RAA assay is a rapid, simple and reliable tool for accurate detection of P. aeruginosa of diverse origins.

Okra is widely used as vegetable, food ingredient, and traditional medicine in China for various applications. The main healthy properties are accepted to originate from the high polysaccharide content of okra. In this study, a homogenous pectic polysaccharide WOP-2 was extracted, isolated, and purified from okra by DEAE-cellulose and Sepharose CL-6B column, and its molecular weight was estimated to be 580 kDa. Monosaccharide composition was analyzed to be Rha (21.4%), GalA (34.9%), Gal (29.6%), GlcA (4.5%), Glc (5.9%), and Ara (3.7%) by HPLC with PMP derivatization. Combined with NMR analysis, WOP-2 was elucidated to be a rhamnogalacturonan I backbone with type-II arabinogalactan side-chains substituted partly at O-4 of Rhap. WOP-2 exhibits significant anti-hyperglycemic activity on STZ-induced diabetic mice by inhibiting the lipid peroxidation chain reaction probably. Overall, these results provide new insight into structure–activity relationships of polysaccharide from okra and provide impetus towards the development of polysaccharide-based therapeutics on diabetes.

Objective This study aimed to investigate serum levels of the cystatin C (CysC)/creatinine (Cr) ratio and renal serum markers (CysC, Cr, urea, and uric acid [UA]) for different ages and by sex. We also aimed to establish pediatric reference intervals for the serum CysC/Cr ratio. Methods Serum samples were collected from 4765 healthy children (0–18 years old). Serum markers of renal function were measured, and the CysC/Cr ratio of each participant was calculated and statistically analyzed. Results The renal marker CysC did not substantially change after 1 year old. Cr, urea, and UA levels generally increased with age. However, the serum CysC/Cr ratio steadily decreased with age. The CysC/Cr ratio showed significant differences in age among all age groups and varied with sex, except for in the 1 to 6-year-old groups. The overall serum CysC/Cr ratio in girls was higher than that in boys. Conclusion Reference intervals of the serum CysC/Cr ratio in the pediatric population were established. These intervals need to be partitioned by age and sex.

Cultivation substrate water status is of great importance to the production of netted muskmelon (Cucumis melo L. var. reticulatus Naud.). A prediction model for the substrate water status would be beneficial in irrigation schedule guidance. In this study, the machine learning random forest model was used to forecast plant substrate water status given the phenotypic traits throughout the muskmelon growing season. Here, two varieties of netted muskmelon, “Wanglu” and “Arus”, were planted in a greenhouse under four substrate water treatments and their phenotypic traits were measured by taking the images within the visible and near-infrared spectrums, respectively. Results showed that a simplified model outperformed the original model in forecasting speed, while it only uses the top five most significant contribution traits. The forecast accuracy reached up to 77.60%, 94.37%, and 90.01% for seedling, vine elongation, and fruit growth stages, respectively. Combining the imaging phenotypic traits and machine learning technique would provide a robust forecast of water status around the plant root zones.

Junggar basin is one of the most important oil and gas exploration blocks in China. Mahu sag, as the most important hydrocarbon generation sag, is rich in oil and gas resources and has good exploration prospects. Taking the reservoir of lower Wuerhe formation in Madong slope belt of Mahu sag as the research object, this paper studies the reservoir characteristics and main controlling factors of lower Wuerhe formation on the basis of core data and logging data. The results show that the main rock type of lower Wuerhe formation is glutenite, and there are four types of reservoir space: primary intergranular pore, residual intergranular pore, intragranular dissolved pore and intergranular dissolved pore. Reservoir porosity ranges from 5.12% to 10.30%, with an average of 7.15%, and permeability ranges from 0.01 to 100mD, with an average of 0.24mD. Reservoir development in this area is mainly controlled by tectonism, sedimentary microfacies, diagenesis and other factors, in which sedimentation is the main controlling factor. Compaction is the main factor that destroys reservoir physical properties, and the nose convex structure formed by tectonic action is the favorable area of structural reservoir. Combined with the drilling logging data and oil test and production data, the favorable area of lower Wuerhe formation in the study area is predicted, and it is pointed out that type I1, type I2 and type II are the dominant reservoirs. The study of reservoir characteristics and main controlling factors of lower Wuerhe formation in Madong slope belt has guiding significance for exploration in the study area. [ABSTRACT FROM AUTHOR]

The Jurassic in the Sanquan bulge of Luliang Oilfield, Junggar Basin, China is rich in oil and gas resources. However, the area is located in a desert area and its geological features are very complex. Therefore, the use of seismic data for reservoir prediction is an effective method. In this study, the three-dimensional seismic data system was used to carry out the fine fault characterization, structural interpretation, reservoir characterization and trap evaluation of the Jurassic strata. The research results show that the use of coherent horizontal slices can effectively realize fault recognition. The fault system of the Sanquan Uplift can be subdivided into two groups, one group is in NE-SW direction, the other group is in NW-SE direction. The study area is dominated by normal faults, which can be subdivided into two stages, the first stage is the early Yanshanian fault, and the other is the middle-late Yanshanian fault. The structural interpretation results show that different provenance sources of the Jurassic Toutunhe Formation have different seismic response characteristics. The source in the northwest direction shows a poorly continuous medium-weak reflection in the seismic axis, while the north-east source shows a superimposed feature and a moderately strong reflection with good continuity in the seismic axis. The argillaceous barrier zone between the two provenances showed blank reflection characteristics. According to regional seismic and geological fine interpretation, sedimentary reservoir research and reservoir profile analysis, we found that the Jurassic trap types are mainly: (1) Toutunhe Formation lithological traps developed on the perimeter; (2) The fault block, nose or fault anticline traps developed on the anticline belt of the Jurassic Xishanyao Formation. Finally, we identified the anticlines and fault anticlines traps in the Toutunhe Formation and the fault block traps in the Xishanyao Formation. [ABSTRACT FROM AUTHOR]