79 results about "Granular leucocyte" patented technology

The invention belongs to the technical field of biological medicine, and particularly relates to a reagent for improving ovarian development and a preparation method and application thereof. The reagent for improving ovarian development is prepared by mixing the following raw materials in parts by weight: 20-30 parts of N-acetylcysteine; 5 to 10 parts of tranexamic acid; 3 to 8 parts of creatine; and 3000 parts of water. According to the reagent, the mRNA expression of PKD1P6 and follicle stimulating hormone receptors (FSHR) in ovarian tissues is up-regulated, the activity and functions of cumulus granular cells are enhanced, and apoptosis of the cumulus granular cells is inhibited, so that the effects of effectively promoting follicle development and inhibiting the apoptosis of the cumulus granular cells are achieved.

The invention relates to the technical field of animal husbandry, in particular to a synchronous oestrus-timing insemination method suitable for cattle, which comprises the following steps: firstly, performing intramuscular injection of PG (cloprostenol sodium) at different time points after calving to promote uterine recovery and luteolysis and lay a synchronous oestrus foundation; the method comprises the key steps that GnRH (gonadotropin release hormone) is given through intramuscular injection on the 43rd day, the 53rd day, the 60th day and the 69th day, pituitary is promoted to release follicle-stimulating hormone (FSH) and luteinizing hormone (LH), collection and screening of follicles are guided, follicle maturation is promoted, the oestrus uncovering rate and the oestrus conception rate are increased, a melatonin implant is implanted in a subcutaneous implantation mode on the 63rd day, and the melatonin implant is implanted in a subcutaneous implantation mode on the 63rd day. The synthesis of ovarian hormones is promoted by regulating the hypothalamus-pituitary-gonad axis, the secretion of follicle cell factors and maturation promoting factors is promoted, the development of granular cells is promoted, and the maturation of oocytes is promoted. According to the method, the pregnancy rate of a double-synchronization-timing insemination scheme which is most widely used in a large-scale dairy cow breeding pasture at present can be remarkably increased, and feasible technical support is provided for success of increasing the breeding efficiency of cows.

This invention provides the application of CTH inhibitors and / or xCT inhibitors in the preparation of drugs for inhibiting mammalian embryo implantation, relating to the field of pharmaceutical technology. This invention verifies the efficacy of cystathionine-γ-lyase (CTH) inhibitors and / or solute carrier family 7 member 11 (xCT) inhibitors in inhibiting mammalian embryo implantation. The results show that xCT inhibitors and / or CTH inhibitors can significantly inhibit the proliferation of luteinized granulosa cells and progesterone secretion in mouse models. Furthermore, in vivo injection of xCT inhibitors and / or CTH inhibitors into mouse models not only significantly inhibits ovulation and the maturation rate of released oocytes, but also significantly reduces the formation of functional corpus luteum and embryo implantation. Therefore, this application provides new ideas and methods for the development of contraceptive drugs.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.

The present application relates to the field of biological medicine and modernization of traditional Chinese medicine, and particularly relates to a therapeutic target, an inhibitor and a traditional Chinese medicine composition for treating ovarian hypofunction caused by oxidative stress and application thereof. The present application first determines SRC tyrosine kinase as a key drug target for preventing and treating the disease. The application of the SRC tyrosine kinase inhibitor (such as secaitinib) in preparing related drugs is provided, which can alleviate the damage of ovarian granulosa cells by inhibiting the SRC activity, down-regulating the expression of antioxidant enzyme related genes and reducing the level of active oxygen. Meanwhile, the present application provides a traditional Chinese medicine composition composed of mulberry, kudzu root, tuckahoe and medlar. The composition can improve the hormone level, enhance the antioxidant capacity, improve the ovarian function and fertility by inhibiting the SRC tyrosine kinase activity and the SRC-RAF1-MEK-ERK signal pathway. The present application provides a new solution for the targeted treatment of ovarian hypofunction and the modernization of traditional Chinese medicine.

The application discloses a kind of marker for predicting oocyte copy number variation and application thereof, the marker includes the combination of CLEC11A, P4HB, EFEMP2, IL32, FTL, FLNA, COL6A3, ACPP and APOO protein.By the marker of the application, the expression level of granulosa cell secretory protein can be determined by granulosa cell transcriptome sequencing, and the genomic CNV condition of corresponding oocyte can be inferred.The detection object of the technical solution is discarded granulosa cell in assisted reproductive technology, so it will not have any impact on oocyte, and it is non-invasive;The present study is a transcriptome and methylation group detection from single cell level, with high-throughput characteristics, and all granulosa cells and oocytes are one-to-one matched, which can reflect the difference between single cell level COCs.

The present invention relates to a medium composition for in vitro fertilization and / or in vitro culture comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a method using same. The compound or composition suppresses reactive oxygen species and lipid peroxidation in ovarian granulosa cells to preventing cytotoxicity and mitochondrial dysfunction caused by apoptosis and ferroptosis, leading to an improvement in the quality of aged oocytes, and to increase the developmental rate and blastocyst formation rate of pre-implantation embryos, thereby improving the efficiency of in vitro fertilization. In addition, treatment with the medium composition during the vitrification process of in vitro–fertilized embryos improves re-expansion and survival rates, thereby reducing structural and metabolic damage. Accordingly, the present invention can be advantageously applied to an assisted reproductive technology for treating infertility and subfertility, and in vitro fertilization and / or in vitro culture for improving the propagation efficiency of livestock.

The invention discloses a method for promoting ovulation of sexual maturity zebrafish by using Gap27, and relates to the technical field of artificial breeding of fish, a gap connecting protein inhibitor Gap27 is applied to ovulation induction of zebrafish, an intercellular communication network in follicles is accurately intervened, the problem of asynchronous ovulation is solved, and in the final stage of maturation of oocytes, ovulation of sexual maturity zebrafish is promoted. Close substance and information exchange is carried out among follicular granule cells through a gap connecting channel formed by connecting protein Cx43, and Gap27 is used as a specific peptide fragment, can simulate an extracellular ring structure of Cx43, and competitively binds with Cx43 on an adjacent cell membrane, so that formation and functions of the gap connecting channel are physically blocked, and the follicular granule cells are separated from each other. When ovarian tissues of sexually mature female fish are exposed in a Gap27 solution, the transmission of inhibitory signals from granular cells to oocytes can be effectively interrupted, the rupture of nuclear membranes of the cells is promoted, and the final maturation process is completed.

The application relates to the field of biological medicine, and particularly discloses application of a sophora japonica extract in preparation of a product for improving the reproductive performance of ewes. The sophora japonica extract can effectively inhibit pyroptosis of ovarian granulosa cells by down-regulating expression of cysteine protease B, further reverses follicle atresia and ovulation disorders caused by pyroptosis, and improves the reproductive performance; animal experiments show that the sophora japonica extract can optimize the follicle development structure of ewes, inhibit overdevelopment of large follicles, and significantly improve the ovulation efficiency, thereby providing a new scheme for inhibiting pyroptosis of ovarian granulosa cells and improving the reproductive performance of livestock.

Provided herein are methods and compositions for differentiating induced pluripotent stem cells into granulosa-like cells by overexpressing transcription factors such as NR5A1 and a RUNX family protein (e.g., RUNXI and / or RUNX2).

The application discloses a doxorubicin scavenger for protecting ovarian fertility and a preparation method thereof, comprising a DNA complex wrapped by a red blood cell membrane, wherein the red blood cell membrane is surface-modified by an anti-Mullerian duct antibody alphaAMH; the doxorubicin scavenger is specifically combined with AMH which is highly expressed in ovarian tissues through alphaAMH, is enriched in the ovarian part in a targeted manner, adsorbs doxorubicin through a DNA fragment in the DNA complex, reduces toxic damage of doxorubicin to ovarian cells, and further protects ovarian fertility. The doxorubicin scavenger can adsorb injected doxorubicin Dox, reduce side effects of doxorubicin Dox on ovarian granulosa cells, and further protect ovarian function and fertility.

The invention relates to an effect of an REV-ERBalpha agonist SR9009 in the aspect of delaying ovarian senescence, and aims at the problem of fertility decline caused by cliff-type decline of ovarian functions of women over 35 years old, the SR9009 is applied to activate an REV-ERBalpha circadian rhythm signal channel in ovarian tissues, inhibit excessive autophagy of granular cells and maintain a primordial follicle dormancy state, so that the agonist SR9009 can be used for delaying ovarian senescence. Therefore, the original follicle depletion speed is delayed, follicle storage is maintained, and the ovary reproductive function cycle is prolonged. The effect is verified by a mammal model: REV-ERBalpha protein expression in an ovary of an aging individual is reduced, rhythmicity expression is disordered, autophagy of granulosa cells can be effectively inhibited in the ovary of the aging individual intervened by the SR9009, and the number of residual primordial follicles and the number of total follicles are obviously greater than those of an untreated control group. The invention provides an innovative treatment strategy of targeted rhythm regulation and control for clinical intervention of ovarian aging.

The present application relates to a kind of two-dimensional metal polyphenol complex nanomaterials that can be enriched in ovary, preparation method and application thereof, the nanomaterial can be enriched in ovary, including two-dimensional metal polyphenol complex as substrate, and follicle-stimulating hormone polypeptide grafted to the surface of the substrate;Wherein, the surface of the two-dimensional metal polyphenol complex grafts-COOH functional group and forms 2D MPC-COOH intermediate, the follicle-stimulating hormone polypeptide is covalently grafted with the-COOH on the surface of 2D MPC-COOH intermediate and obtains the two-dimensional metal polyphenol complex nanomaterial.The technical effect of the present application is that the preparation method of nanomaterial is simple, and nanoparticle dispersibility is good, and biosafety is high, and nanomaterial can physically absorb BRD7 target protein through its special structure, block BRD7 into cell nucleus, and then realize the safe down-regulation of KAT7 path and the effective reversal of granulosa cell stress aging.

The application discloses a molecular genetic marker affecting pig sexual maturity and application thereof. The application takes RBP1 as a research object, and adopts molecular and cell biology methods to study the correlation between RBP1 and the initial estrus period. It is found that six SNP sites in the RBP1 promoter region are related to the initial estrus period. Further research shows that the promoter activity of the CC genotype of the g.80408109T>C site is significantly higher than that of the TT genotype, the promoter activity of the AA genotype of the g.80409632A>G site is significantly higher than that of the GG genotype, and it is confirmed that RBP1 can inhibit the ferroptosis of granulosa cells. It can be seen that the mutation in the RBP1 promoter region can affect the expression of RBP1, and RBP1 can accelerate the initial estrus start by inhibiting the ferroptosis of pig ovarian granulosa cells. The application has good application value for the research on ovarian follicle development and initial estrus start.

The invention discloses an ergothioneine composition for improving ovarian vitality and a preparation method and application thereof, the composition comprises D-ribose and ergothioneine, the concentration of the D-ribose in the composition is 5-15 mM, and the concentration of the ergothioneine in the composition is 100-300 [mu] M. The pharmaceutical composition is used for regulating the redox balance of ovarian granular cells, inhibiting inflammatory response and up-regulating anti-mullerian hormone secretion so as to improve the ovarian activity.

This invention discloses a Polygonatum sibiricum vesicle preparation for treating premature ovarian failure, its preparation method, and its application. The preparation method includes: cleaning Polygonatum sibiricum, juicing, filtering to remove residue, and collecting the juice; subjecting the juice to gradient centrifugation to remove residual plant tissue, plant cells, and plant cell debris; then using an extraction reagent for stepwise precipitation, collecting the precipitate, and resuspending it to obtain the final product. The Polygonatum sibiricum vesicle preparation prepared by this invention can be efficiently taken up by ovarian granulosa cells. It effectively treats premature ovarian failure by activating the Nrf2 signaling pathway, upregulating antioxidant enzyme expression, scavenging reactive oxygen species, restoring mitochondrial membrane potential, inhibiting apoptosis, and restoring the expression levels of estrogen, follicle-stimulating hormone, and anti-Müllerian hormone. The preparation method of this invention is standardized, biocompatible, and safe, and has broad clinical application prospects.

The invention belongs to the technical field of biological medicines, and particularly relates to application of an SIRT5 inhibitor MC3482 in preparation of a medicine for treating premature ovarian failure. According to the invention, a mouse POI model is constructed through the induction of cyclophosphamide (CTX), and the MC3482 is proved to be capable of inhibiting the activity of SIRT5, reducing excessive macrophage infiltration of ovary caused by CTX, reducing M1 type macrophage activation and proinflammatory factor release, further inhibiting the apoptosis of ovarian granular cells, protecting follicular reserve and recovering ovarian functions. The invention clarifies the action effect and application scheme of MC3482 for preventing and treating POI, provides a new treatment strategy for chemotherapy-induced iatrogenic POI, and has important clinical transformation value.

Featured are methods, compositions, and apparatuses for the in vitro maturation of oocytes. In particular, the disclosure features methods of inducing oocyte maturation in vitro, by co-culturing a female subject's oocytes with an ex vivo composition containing a plurality of ovarian support cells (e.g., granulosa cells). Additional methods for administering follicular triggering agents and retrieving oocytes from the female subject are provided. Such methods, compositions, and apparatuses are particularly useful for assisted reproduction technology (ART) procedures.

The invention discloses an application of butyric acid or a derivative thereof in preparation of a zearalenone reproductive toxicity resisting preparation. Also disclosed is an antitoxic feed composition comprising a specific base ration and a butyric acid derivative. In-vivo and in-vitro tests and molecular mechanism researches prove that butyric acid and derivatives thereof (such as tributyrin and sodium butyrate) can effectively relieve zearalenone (ZEN)-induced animal ovary injury. The action mechanism of the traditional Chinese medicine composition is that follicular atresia is reduced, the balance of reproductive hormones (such as GnRH, E2, AMH and P4) is recovered, and clinical symptoms caused by ZEN poisoning such as vulva swelling and the like are relieved by activating a BMPs / SMAD signal channel, especially up-regulating SMAD4 protein expression and inhibiting oxidative stress (ROS) and Caspase-3 mediated granular cell apoptosis.

The application discloses application of an IRS1 gene promoter region SNP site as a pig sexual maturity molecular genetic marker. The present application takes the IRS1 gene promoter region as a research object, and studies the correlation between the IRS1 gene promoter region and sow sexual maturity by using molecular and cell biology methods: it is found that g.128307611C>A and g.128307308C>T are significantly correlated with the age of sexual maturity of Duhei pig populations; then, the influence of different genotypes of the above sites on the activity of the IRS1 gene promoter is studied by transfecting different vectors. Further, the sow ovarian granulosa cells are taken as experimental materials, and experimental techniques such as transcriptome sequencing, total iron colorimetry and malondialdehyde colorimetry are used to study the influence of the IRS1 gene on the transcription level and ferroptosis of the sow ovarian granulosa cells, and it is found that the IRS1 gene influences the different genes of the cell transcriptome, and is mainly enriched in signal pathways such as iron ion homeostasis, and inhibits the ferroptosis of the cells.

The application relates to a method for establishing an early ovarian failure rat model, which comprises the following steps: taking healthy rats, and performing gavage on the rats according to a dosage of 10-20 mg / Kg of a phytostilbene preparation, wherein the dosage is the ratio of phytostilbene to the weight of the rats, and obtaining the early ovarian failure rat model after four days of administration. The early ovarian failure rat model is obtained by performing gavage on the rats after the phytostilbene preparation, the model is simple to establish, and the model establishment time is short. Through ovarian pathological observation, it can be seen that the simulated animal model has obvious ovary tissue damage, follicular granulosa cell destruction and apoptosis, and follicle atresia and other symptoms. The obtained model has good stability and repeatability. The phytostilbene is used to successfully establish the early ovarian failure rat model, lays a good foundation for establishing a stable early ovarian failure model for basic research, and provides a reliable method for the construction of the early ovarian failure model.

The invention relates to the technical field of livestock breeding biology, and discloses a method for promoting porcine ovary granular cell proliferation, which comprises the following steps: adding a porcine follicular fluid exosome into a porcine ovary granular cell culture system, and acting for a certain time at an effective concentration, so as to improve the proliferation activity of granular cells. After the exosome treatment, the apoptosis rate of granular cells can be reduced, the cell survival rate can be improved, and the granular cells can be promoted to secrete steroid hormones, including the estradiol level and the expression of progesterone synthetase. Cell experiments prove that the follicular fluid exosome can effectively improve the activity of granular cells, promote the process of a cell cycle, inhibit cell apoptosis and remarkably enhance the synthesis capacity of steroid hormones such as estradiol and progesterone, and a new intervention strategy is provided for improving follicular development and reproductive performance of sows.

The invention provides an application of an E3 ubiquitin ligase RLIM gene inhibitor in preparation of a medicine for treating ovarian aging or injury. Experiments find that the expression level of the RLIM gene in the ovary is positively correlated with the aging degree, and the expression of the RLIM gene is remarkably improved along with age increase. In-vivo and in-vitro experiments show that ovarian granular cell senescence can be effectively delayed and ovarian functions can be improved by inhibiting RLIM expression, and particularly, the proportion of atresia follicles is reduced, the number of healthy follicles is increased, and serum AMH and E2 levels are remarkably improved. The pharmaceutical composition provided by the invention comprises a siRNA inhibitor (preferably, siRLIM-2) of specific targeting RLIM and a nano-liposome carrier, and efficient targeting delivery can be realized through tail vein injection. The treatment scheme has no toxic effect on important organs, is high in safety, and provides a new effective means for treatment of ovarian aging and related diseases.

This invention relates to the field of biomedical technology and discloses a biomarker for diagnosing polycystic ovary syndrome (PCOS) and its application. The diagnostic biomarker is circSPECC1(4), which is formed by reverse splicing and circularization of the fourth exon of the SPECCC1 gene. It has a length of 1580 nt and possesses a closed circular RNA structure. circSPECC1(4) is specifically highly expressed in ovarian granulosa cells of PCOS patients, and there is no significant difference in the mRNA expression level of its parent gene SPECCC1. circSPECC1(4) possesses RNase R nuclease resistance stability and is mainly located in the cytoplasm. Knocking down the expression of circSPECC1(4) can significantly promote apoptosis of ovarian granulosa cells and arrest the ovarian granulosa cell cycle at the G0 / G1 phase. This invention clearly demonstrates that circSPECC1(4) can serve as a specific diagnostic biomarker for PCOS, fully verifying the structural characteristics, expression specificity, and pathological regulatory function of this circular RNA, filling the gap in the existing field of PCOS diagnosis which lacks highly specific and stable molecular diagnostic targets.

The invention relates to the technical field of biological medicines, and particularly discloses application of formononin in preparation of a medicine for regulating and controlling BNIP3-mediated excessive mitochondrial autophagy. The invention discloses that ononin can be used as a direct small molecule inhibitor of BNIP3 protein, specific binding of ononin with BNIP3 is verified through molecular docking, surface plasmon resonance and molecular dynamics simulation in multiple dimensions, and in an iron overload induced ovarian granulosa cell injury model, the ononin can be used as an iron overload induced ovarian granulosa cell injury model. The formononin effectively inhibits mitochondrial excessive autophagy mediated by a BNIP3-PINK1 / Parkin axis through targeting BNIP3, remarkably recovers mitochondrial membrane potential, reduces mitochondrial active oxygen level and improves cell oxygen consumption rate (OCR), so that stressful high-flux and low-efficiency breathing states are corrected, cell senescence phenotypes are delayed, and cell apoptosis is inhibited. And a new small molecule drug strategy with a clear structure and a clear action mechanism is provided for iron overload related ovarian function impairment.

This invention discloses a method for the isolation, culture, and identification of chicken sexual gonadal cells, belonging to the field of biotechnology. The invention selects chicken embryos hatched to 18.5 days old, isolates the gonads after PCR sex identification, and uses 0.25% trypsin-EDTA digestion followed by filtration through a 70μm filter to obtain a single-cell suspension with a survival rate >90% and free from PGC contamination. A sex-specific culture system is established, allowing cells to be stably passaged and maintain a sex-specific phenotype. Through morphological, qRT-PCR, immunofluorescence, and Western blot multidimensional identification, accurate identification of male testicular Sertoli cells and female ovarian granulosa cells is achieved. This method is simple to operate, has good reproducibility, and produces high cell viability and purity. It can provide a stable gonadal microenvironment for the in vitro directed differentiation of chicken primordial germ cells, significantly improving gamete induction efficiency. It is suitable for research on avian reproductive and developmental mechanisms, transgenic breeding, and germplasm resource preservation.

This invention belongs to the field of biomedical technology, specifically relating to the application of a composition in the preparation of a drug for the prevention and treatment of polycystic ovary syndrome (PCOS). The composition comprises bovine spleen peptide and adenosine; the mass ratio of bovine spleen peptide to adenosine is 1:(1-2.5). Bovine spleen peptide, as a bioactive polypeptide, significantly improves insulin resistance associated with PCOS by targeting and activating the PI3K-Akt signaling pathway and promoting insulin receptor substrate phosphorylation. Adenosine, as a natural bioactive substance, significantly reduces serum luteinizing hormone and testosterone levels in rats and increases follicle-stimulating hormone levels, thereby effectively alleviating hyperandrogenism symptoms such as hirsutism and acne in PCOS. Furthermore, this composition can significantly improve polycystic pathological changes in the ovarian tissue of model rats, reduce the number of cystic dilated follicles, restore the granulosa cell layer structure, alleviate ovarian stromal hyperplasia, and promote normal follicular development.

The invention belongs to the technical field of biological medicine, and particularly relates to application of irisin in treatment of polycystic ovarian syndrome, and the specific application of irisin in treatment of polycystic ovarian syndrome is as follows: improving the ovarian morphology and ovarian dysfunction of a patient suffering from PCOS (polycystic ovarian syndrome); the ovarian ferroptosis and iron autophagy of the PCOS patient are improved; ferroptosis of primary ovarian granular cells and KGN cells induced by high androgen is improved; iris is used for regulating and controlling an NCOA4-FTH pathway to reverse iron autophagy of ovarian granulosa cells, and through a PCOS mouse model induced by hyperandrogen, specific targets of irisin for inhibiting ferroptosis of the ovarian granulosa cells are disclosed, abnormal iron metabolism of PCOS patients is reversed, and the normal ovulation function of the PCOS patients is recovered; a safe alternative is provided: irisin can be generated endogenously by movement, avoiding the risk of metabolic disorder caused by exogenous hormone therapy.

The invention relates to an application of an HK1 gene in mouse ovarian granular cells, in particular to an application in regulating proliferation and apoptosis of the mouse ovarian granular cells and / or steroid hormone synthesis. According to the invention, mouse ovarian granular cells are taken as a research object, and the expression of the HK1 gene is interfered to verify that the HK1 gene is taken as a key target spot for regulating and controlling the functions of the mouse ovarian granular cells so as to influence steroid hormone synthesis and cell proliferation and apoptosis processes. The invention not only provides more reliable technical support for ovarian function regulation, but also provides new thinking and technical guidance for mechanism research of reproductive diseases and preparation and development of related drugs.