Effect of Vibration and Tilting Mechanical Loading Device on Parthenogenetic Embryos Development of Mice Immature Oocytes.

The growth and development of early mammalian embryos mainly take place in the fallopian tube, which not only provides nutrients for embryonic growth and development but also offers suitable mechanical conditions. The embryo culture system established in assisted reproductive technology mainly simulates the environment in which oocytes and embryos grow and develop in vivo. However, current in vitro embryo culture is mainly static and cannot completely mimic the mechanical environment in which embryos grow and develop in vivo. Therefore, to more accurately simulate the mechanical environment of embryos in the fallopian tube, we have developed a dynamic culture device to investigate the effects of mechanical stimulation on the in vitro maturation of immature oocytes and their parthenogenetic developmental potential. Immature mice oocytes were subjected to in vitro maturation by static culture and vibration (3 Hz, 6 Hz) with tilting for 15∼16 hours. The maturation of oocytes was observed after the culture period. The mature oocytes were activated by parthenogenesis and the rate of embryo compaction and formation of parthenogenetic blastocysts was analyzed. The results showed that using 3 Hz vibration and tilting can significantly improve the parthenogenetic development potential of immature mice oocytes.

Effect of Mechanical Stimulations on the Cytoskeleton of Mouse Oocytes During Maturation.

In the reproductive system of female mammals, the early embryos grow and develop in the fallopian tube, where they are stimulated by fluid flow and ciliary vibration. The mechanical environment of the fallopian tube affects the development of embryos. This study is focused on the role of mechanical stimulation on the cytoskeleton of oocytes during oocyte maturation in vitro. The 3 Hz microvibration and tilting stimulations were applied to mouse immature oocytes. The oocyte maturation rate and area of the first polar body under dynamic stimulation were compared with those of the static culture group. A fluorescence assay was used to study the localization of the cytoskeleton during oocyte maturation. The results indicated that mechanical stimulation can change the localization of F-actin and spindle, and oocytes discharged smaller areas of the first polar body.

Detection of the effect of microvibrational stimulation on human discarded immature oocytes by single-cell transcriptome sequencing technology.

OBJECTIVE: This study aimed to investigate the changes in oocytes at the transcriptome level after applying continuous microvibrational mechanical stimulation to human immature oocytes during in vitro maturation. METHODS: The discarded germinal-vesicle stage (GV) oocytes with no fertilization value after oocytes retrieval in assisted reproduction cycles were collected. Part of them was stimulated with vibration (n = 6) at 10 Hz for 24 h after obtaining informed consent; the other was cultured in static condition (n = 6). Single-cell transcriptome sequencing was used to detect the differences in oocyte transcriptome compared with the static culture group. RESULTS: The applied 10-Hz continuous microvibrational stimulation altered the expression of 352 genes compared with the static culture. Gene Ontology (GO) analysis suggested that the altered genes were mainly enriched with 31 biological processes. The mechanical stimulation upregulated 155 of these genes and downregulated 197 genes. Among them, the genes related to mechanical signaling, such as protein localization to intercellular adhesion (DSP and DLG-5) and cytoskeleton (DSP, FGD6, DNAJC7, KRT16, KLHL1, HSPB1, MAP2K6), were detected. DLG-5, which was related to protein localization to intercellular adhesion, was selected for immunofluorescence experiments based on the transcriptome sequencing results. The protein expression of DLG-5 in the microvibration-stimulated oocytes was higher than that in the static culture oocytes. CONCLUSIONS: Mechanical stimulation affects the transcriptome during oocyte maturation, causing the express changes in intercellular adhesion and cytoskeleton-related genes. We speculate that the mechanical signal may be transmitted to the cell through DLG-5 protein and cytoskeleton-related protein to regulate cellular activities.

A multitask model for realtime fish detection and segmentation based on YOLOv5.

The accuracy of fish farming and real-time monitoring are essential to the development of "intelligent" fish farming. Although the existing instance segmentation networks (such as Maskrcnn) can detect and segment the fish, most of them are not effective in real-time monitoring. In order to improve the accuracy of fish image segmentation and promote the accurate and intelligent development of fish farming industry, this article uses YOLOv5 as the backbone network and object detection branch, combined with semantic segmentation head for real-time fish detection and segmentation. The experiments show that the object detection precision can reach 95.4% and the semantic segmentation accuracy can reach 98.5% with the algorithm structure proposed in this article, based on the golden crucian carp dataset, and 116.6 FPS can be achieved on RTX3060. On the publicly available dataset PASCAL VOC 2007, the object detection precision is 73.8%, the semantic segmentation accuracy is 84.3%, and the speed is up to 120 FPS on RTX3060.

Effects of microvibration stimulation on developmental potential of discarded germinal vesicle oocytes of human.

Objective: This research aims to study the effects of continuous microvibration stimulation on the parthenogenetic development of human germinal vesicle oocytes. Methods: Ninety-five discarded germinal vesicle oocytes from intracytoplasmic sperm injection treatment (ICSI) cycles performed at Amcare Women's & Children's Hospital between January and December 2021 were used for conventional static culture as well as 10 Hz microvibration culture. We investigated the differences between the two groups in terms of oocyte maturation rate, parthenogenetic activation rate, and parthenogenetic blastocyst formation rate. Results: The static culture and 10 Hz microvibration culture of 95 oocytes showed that the parthenogenetic blastocyst formation rate in the microvibration culture group was significantly higher than those in the traditional static culture group. Conclusion: A continuous microvibration stimulation can significantly improve the parthenogenetic developmental potential of human immature oocytes.

Study of a QueryPNet Model for Accurate Detection and Segmentation of Goose Body Edge Contours.

With the rapid development of computer vision, the application of computer vision to precision farming in animal husbandry is currently a hot research topic. Due to the scale of goose breeding continuing to expand, there are higher requirements for the efficiency of goose farming. To achieve precision animal husbandry and to avoid human influence on breeding, real-time automated monitoring methods have been used in this area. To be specific, on the basis of instance segmentation, the activities of individual geese are accurately detected, counted, and analyzed, which is effective for achieving traceability of the condition of the flock and reducing breeding costs. We trained QueryPNet, an advanced model, which could effectively perform segmentation and extraction of geese flock. Meanwhile, we proposed a novel neck module that improved the feature pyramid structure, making feature fusion more effective for both target detection and instance individual segmentation. At the same time, the number of model parameters was reduced by a rational design. This solution was tested on 639 datasets collected and labeled on specially created free-range goose farms. With the occlusion of vegetation and litters, the accuracies of the target detection and instance segmentation reached 0.963 (mAP@0.5) and 0.963 (mAP@0.5), respectively.

Research on remote sensing image extraction based on deep learning.

Remote sensing technology has the advantages of fast information acquisition, short cycle, and a wide detection range. It is frequently used in surface resource monitoring tasks. However, traditional remote sensing image segmentation technology cannot make full use of the rich spatial information of the image, the workload is too large, and the accuracy is not high enough. To address these problems, this study carried out atmospheric calibration, band combination, image fusion, and other data enhancement methods for Landsat 8 satellite remote sensing data to improve the data quality. In addition, deep learning is applied to remote-sensing image block segmentation. An asymmetric convolution-CBAM (AC-CBAM) module based on the convolutional block attention module is proposed. This optimization module of the integrated attention and sliding window prediction method is adopted to effectively improve the segmentation accuracy. In the experiment of test data, the mIoU, mAcc, and aAcc in this study reached 97.34%, 98.66%, and 98.67%, respectively, which is 1.44% higher than that of DNLNet (95.9%). The AC-CBAM module of this research provides a reference for deep learning to realize the automation of remote sensing land information extraction. The experimental code of our AC-CBAM module can be found at https://github.com/LinB203/remotesense.

Identification of two novel poleroviruses and the occurrence of Tobacco bushy top disease causal agents in natural plants.

Tobacco bushy top disease (TBTD) is a devastating tobacco disease in the southwestern region of China. TBTD in the Yunnan Province is often caused by co-infections of several plant viruses: tobacco bushy top virus (TBTV), tobacco vein distorting virus (TVDV), tobacco bushy top virus satellite RNA (TBTVsatRNA) and tobacco vein distorting virus-associated RNA (TVDVaRNA). Through this study, two new poleroviruses were identified in two TBTD symptomatic tobacco plants and these two novel viruses are tentatively named as tobacco polerovirus 1 (TPV1) and tobacco polerovirus 2 (TPV2), respectively. Analyses of 244 tobacco samples collected from tobacco fields in the Yunnan Province through RT-PCR showed that a total of 80 samples were infected with TPV1 and/or TPV2, and the infection rates of TPV1 and TPV2 were 8.61% and 29.51%, respectively. Thirty-three TPV1 and/or TPV2-infected tobacco samples were selected for further test for TBTV, TVDV, TBTVsatRNA and TVDVaRNA infections. The results showed that many TPV1 and/or TPV2-infected plants were also infected with two or more other assayed viruses. In this study, we also surveyed TBTV, TVDV, TBTVsatRNA and TVDVaRNA infections in a total of 1713 leaf samples collected from field plants belonging to 29 plant species in 13 plant families and from 11 provinces/autonomous regions in China. TVDV had the highest infection rates of 37.5%, while TVDVaRNA, TBTV and TBTVsatRNA were found to be at 23.0%, 12.4% and 8.1%, respectively. In addition, TVDV, TBTV, TBTVsatRNA and TVDVaRNA were firstly detected of co-infection on 10 plants such as broad bean, pea, oilseed rape, pumpkin, tomato, crofton weed etc., and 1 to 4 of the TBTD causal agents were present in the samples collected from Guizhou, Hainan, Henan, Liaoning, Inner mongolia and Tibet autonomous regions. The results indicated that TBTD causal agents are expanding its host range and posing a risk to other crop in the field.

Multi-Phase Equilibrium and Solubilities of Aromatic Compounds and Inorganic Compounds in Sub- and Supercritical Water: A Review.

Supercritical water oxidation (SCWO), as a novel and efficient technology, has been applied to wastewater treatment processes. The use of phase equilibrium data to optimize process parameters can offer a theoretical guidance for designing SCWO processes and reducing the equipment and operating costs. In this work, high-pressure phase equilibrium data for aromatic compounds+water systems and inorganic compounds+water systems are given. Moreover, thermodynamic models, equations of state (EOS) and empirical and semi-empirical approaches are summarized and evaluated. This paper also lists the existing problems of multi-phase equilibria and solubility studies on aromatic compounds and inorganic compounds in sub- and supercritical water.

Role of tripartite motif protein 27 as a gametogenesis-related protein in human germ cells.

BACKGROUND: The distribution and functional integrity of members of the tripartite motif (TRIM) protein family are essential for cell proliferation, development and apoptosis, and TRIM proteins have been linked to various cancers. To explore the diagnostic potential and mechanisms of TRIM27 in human spermatogenesis and oogenesis, we analyzed its localization pattern and putative roles in human testes and ovaries. METHODS: TRIM27 mRNA and protein levels in human testes and ovaries were investigated using RT-PCR and western blotting, respectively. TRIM27 was abundantly transcribed in human testes and ovaries, particularly during the early stages of spermatogenesis, and localized in the nuclei of primary spermatocytes. Immunofluorescence also revealed a diffuse distribution in the cytoplasm of round spermatids, and the protein was abundant in ovary tissue during various stages of oogenesis development. RESULTS: TRIM27 mRNA and protein was abundantly transcribed in male and female human germ cells by RT-PCR and western blotting in the human testes followed by the ovary. Immunohistochemical results revealed TRIM27 protein was abundant in the sex body of primary spermatocytes undergoing meiotic prophase during the first cycle of spermatogenesis. Moreover, Trim27 was diffusely localized in the cytoplasm of spermatids and round spermatids. Furthermore, TRIM27 was localized to both the nucleus and cytoplasm of human ovary cells. CONCLUSIONS: TRIM27 as a gametogenesis-related protein could play multiple roles in the regulation of sex body formation and germ cell proliferation during spermatogenesis and oogenesis. The identification and characterization of TRIM27 enhances our understanding of the molecular mechanisms underpinning its functions, and provides insight into its potential role in the pathogenesis of germ cell differentiation and infertility.

Supercritical Fluid Extraction of Metal Chelate: A Review.

Supercritical fluid extraction (SFE), as a new green extraction technology, has been used in extracting various metal species. The solubilities of chelating agents and corresponding metal chelates are the key factors which influence the efficiency of SFE. Other main properties of them such as stability and selectivity are also reviewed. The extraction mechanisms of mainly used chelating agents are explained by typical examples in this paper. This is the important aspect of SFE of metal ions. Moreover, the extraction efficiencies of metal species also depend on other factors such as temperature, pressure, extraction time and matrix effect. The two main complexation methods namely in-situ and on-line chelating SFE are described in detail. As an efficient chelating agent, tributyl phosphate-nitric acid (TBP-HNO3) complex attracts much attention. The SFE of metal ions, lanthanides and actinides as well as organometallic compounds are also summarized. With the proper selection of ligands, high efficient extraction of metal species can be obtained. As an efficient sample analysis method, supercritical fluid chromatography (SFC) is introduced in this paper. Recently, the extraction method combining ionic liquids (ILs) with supercritical fluid has been becoming a novel technology for treating metal ions. The kinetics related to SFE of metal species is discussed with some specific examples.

Impact of hepatitis B virus carrier serostatus on neonatal outcomes after IVF-ET.

An increasing number of infertile, hepatitis B virus-infected individuals have opted for assisted reproductive technology. However, the impact of the hepatitis B virus carrier serostatus on neonatal outcomes has not been evaluated. Data from 504 patients who delivered singletons were analyzed. In females, hepatitis B surface antigen, hepatitis B e antigen, and antibody to hepatitis core antigen seropositivity significantly decreased the gestational age at delivery. In contrast, the male hepatitis B virus serostatus did not affect the gestational age at delivery. Multiple linear regression analysis showed that maternal weight, gestational age at delivery, and infant gender were significantly related to birth weight. The present retrospective study showed that in females, hepatitis B surface antigen, hepatitis B e antigen, and antibody to hepatitis core antigen seropositivity was significantly associated with the gestational age at delivery after in vitro fertilization-embryo transfer (IVF-ET).

Global transcriptome analysis of peripheral blood identifies the most significantly down-regulated genes associated with metabolism regulation in Klinefelter syndrome.

The molecular pathogenesis of Klinefelter Syndrome (KS) is not fully understood. The aim of this study was to determine differences in gene expression patterns between KS patients and control individuals to help identify disease-related genes and biological pathways. Gene expression profiles of five KS patients and five healthy men were determined by microarray; 21 differentially expressed genes with a fold-change >1.5 and q-value <0.05 were identified between the groups. Genes associated with metabolism regulation and encoding liver fatty acid-binding protein (FABP1), aldehyde dehydrogenase 1 family member L1 (ALDH1L1), and vitronectin (VTN) were the most-significantly down-regulated in KS, as confirmed by quantitative reverse transcription PCR. Notably, none of these differentially expressed genes are normally found on the X chromosome. Thus, our results indicate that aberrant metabolism is involved in the pathogenesis of KS. Further elucidation of the how aberrant expression of metabolism-related genes affect the pathogenesis of KS may lead to the development of novel preventative and therapeutic strategies.

The safety of long-term cryopreservation on slow-frozen early cleavage human embryos.

OBJECTIVE: To evaluate the impact of cryopreservation storage time on cleavage-stage embryo survival rate, pregnancy rate, implantation rate, singleton birth weight, and live birth rate. METHODS: This study was a retrospective analysis, including 867 thaw cycles and 3,367 embryos. Women who underwent IVF-FET cycles between 2005 and 2012 were analyzed. The patients were divided into four groups, as follows: group 1 (12-23 months); group 2 (24-35 months); group 3 (36-48 months); and group 4 (≥48 months). RESULTS: The storage time did not have a significant effect on survival, damage rate of the blastomeres, implantation rate, pregnancy rate, singleton birth weight, and live birth rate for embryos frozen at cleavage stages. CONCLUSION: Storage time did not influence the survival and pregnancy outcomes of slow-frozen early cleavage human embryos. The developmental potential of cryopreserved human embryos with different storage times does not appear to have a negative influence on further development.

Spreading area and shape regulate apoptosis and differentiation of osteoblasts.

The in vivo observations have indicated that at the remodeling sites of bone, the spreading area or shape of preosteoblasts is confined by the mineralized matrix. But it remains unknown whether this spreading confinement regulates the differentiation or apoptosis of osteoblasts. In the present study, osteoblast-like cells (MC3T3-E1) were seeded on micropatterned islands with different area and shape. The expression of three osteogenic differentiation markers was measured by immunofluorescence staining and apoptotic cells were detected using a terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labelling assay kit. The membrane fluorescence staining results showed that the actual spreading area of micropatterned osteoblasts coincided with the designed value. When the area of a micropatterned cell was confined as 314 or 615 µm(2), which was lower than that of freely spreading osteoblasts, the circular shape promoted the expression of osteogenic differentiation markers and the percentage of apoptotic osteoblasts compared with the branched shape. This shape-regulated differentiation and apoptosis of osteoblasts with confined spreading area were abolished when actin polymerization was inhibited by cytochalasin D. The present study gives an insight into the roles of spreading morphology on osteoblastic differentiation and apoptosis.