26 results about "Testis tissue" patented technology

The invention discloses a method for separating and transplanting spermatogonial stem cells of Chinese sturgeons, and belongs to the technical field of separation of germline stem cells of fishes. The separation method comprises the following steps: washing Chinese sturgeon testis tissues with an L-15 culture medium containing penicillin / streptomycin double antibodies, and carrying out physical shearing, digestive enzyme enzymolysis and screen filtration to obtain a Chinese sturgeon testis cell suspension; performing percoll density gradient centrifugation to obtain high-purity spermatogonial stem cells of the Chinese sturgeons; and dyeing, washing, resuspending and carrying out micro-injection transplantation to enter the gonad primordium reproductive ridge part of the juvenile acceptor fish. According to the physical characteristics of the spermatogonial stem cells of the Chinese sturgeons, the spermatogonial stem cells with high purity are obtained by adopting percoll density gradient centrifugation, and the purpose of abdomen borrowing reproduction is achieved by adopting a microinjection mode, so that the preservation of germ cells of endangered fishes and the reconstruction of artificial populations are realized; and a new technical approach is provided for protecting genetic resources of the Chinese sturgeons and breaking through artificial propagation bottlenecks.

This invention discloses a method for extracting high-purity PDRN from salmon testes and its application in improving skin condition, belonging to the field of bioactive ingredient extraction technology. Frozen salmon testis tissue is cleaned of blood vessels and fatty impurities, then cryogenically pulverized at -20~-25℃. The pulverized tissue is added to a pre-treatment dispersion buffer and homogenized under high pressure to obtain a DNA fragmentation solution. The precipitate is then separated by centrifugation. The precipitate is dispersed in a dedicated enzyme digestion buffer, and enzymatic digestion with protease and RNase A is performed to inactivate the enzyme. DNase I is added to the digestion solution for enzymatic cleavage. After the reaction, a stop solution is added to terminate the reaction, and the supernatant is separated by centrifugation. Ethanol is added to the supernatant for precipitation, and centrifugation yields PDRN from salmon testes. This invention, through specific buffer systems and enzymatic digestion conditions, increases the PDRN content by 40~60 kDa, allowing for better absorption and utilization by the skin, improving facial hydration and elasticity, effectively reducing facial wrinkles, and improving facial skin condition. It can be used to prepare functional cosmetics.

The invention discloses Lap3a, Lap3b and Lap3c genes of bactrocera cucurbitae and an application of the Lap3a, Lap3b and Lap3c genes. The space-time and tissue expression profiles of the Lap3a, Lap3b and Lap3c genes of the bactrocera cucurbitae show that the three genes are specifically and highly expressed in testis of male bactrocera cucurbitae, especially in transformation regions in testis tissues of adult bactrocera cucurbitae. The Lap3a, Lap3b and Lap3c genes play important functions in the process of participating in maintaining the male reproduction of the bactrocera cucurbitae, when the Lap3a, Lap3b or Lap3c genes of male bactrocera cucurbitae are inhibited, the number of male sperms of the bactrocera cucurbitae is reduced, the reproductive power of the male bactrocera cucurbitae is reduced, the treated male bactrocera cucurbitae is mated with normal female bactrocera cucurbitae, and the egg laying amount and egg hatching rate of the female bactrocera cucurbitae are reduced. The three genes are expected to become SIT potential targets to be applied to pest control, and a new thought is provided for green control of bactrocera cucurbitae.

The invention relates to the technical field of biology, in particular to an extraction process and application of low-endotoxin salmon PDRN, and the extraction process comprises the following steps: S1, treating salmon testis tissue, performing enzymolysis, adding a surfactant and a chelating agent, performing centrifugation, and collecting supernate; s2, breaking a DNA (Deoxyribose Nucleic Acid) fragment; s3, splitting by using restriction endonuclease, filtering after redissolving, and collecting filtrate; s4, adjusting the pH value, adding modified activated carbon, filtering, collecting filtrate, and performing ultrafiltration; and S5, washing, drying and crushing to obtain the PDRN. The PDRN obtained by the extraction process disclosed by the invention is relatively high in yield and purity, low in endotoxin content and wide in application range.

An embodiment relates to polydeoxyribonucleotide (PDRN)-containing exosomes, a composition including same, and a preparation method therefor, wherein the PDRN-containing exosomes of the embodiment are derived from a fish testis tissue and include high-purity, low-molecular weight PDRN. In addition, exosomes containing a significant amount of low molecular weight PDRN can be induced through a simple process of ultrasonic treatment. Furthermore, application of ultrasound to the separated exosomes can prepare PDRN exosomes that are made to have low molecular weights, whereby the exosomes exhibit high intracellular uptake rates and thus can be advantageously used for manufacturing a tissue regeneration or anti-inflammatory composition.

This invention discloses a method for preparing sodium DNA from salmon sperm and its application, belonging to the field of biomedical technology. The preparation method includes the following steps: S1. Raw material pretreatment: Frozen salmon or salmon testis tissue is lysed at -30 to -20℃, then a mixed solution containing Tris-HCl buffer, NaOH, and NaCl is added, followed by the addition of a chitosan-embedded complex enzyme with a total enzyme activity ≥350KU / L. The enzyme is then hydrolyzed at 37±1℃ for 26-30 hours, maintaining a pH of 7.8-8.0 to obtain the enzymatic hydrolysate; S2. DNA synergistic lysis: The pH of the enzymatic hydrolysate is adjusted to 7.8, and the mixture is synergistically treated in an ice-water bath at ≤10℃ by alternating cycles of 25-35kHz variable frequency ultrasound and 100-120MPa high-pressure homogenization for 40 minutes to obtain the DNA lysate. This method solves the problems of high protein residue, uneven molecular weight distribution, low transdermal absorption, high cost, and poor environmental friendliness in traditional extraction processes, and has the advantages of high product purity, strong activity, low cost, and green environmental protection.

The application discloses a method for synchronously marking germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus, which comprises the following steps: cloning MIR17HG-a (long non-coding RNA) of Paralichthys olivaceus by using a probe primer, obtaining a target plasmid after plasmid extraction, carrying out double enzyme digestion and purification to obtain an enzyme-digested plasmid as a probe template, synthesizing Paralichthys olivaceus MIR17HG-a probe by using an SP6 / T7 RNA polymerase, preparing a tissue slice from the testis tissue of Paralichthys olivaceus, and carrying out fluorescent in situ hybridization on the tissue slice by using the Paralichthys olivaceus MIR17HG-a probe, so that the Paralichthys olivaceus MIR17HG-a probe can synchronously mark the germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus. The method solves the defects of traditional marking technology, such as the need for combined marking or multiple step-by-step marking, and fills the blank of synchronous marking of germ cells and interstitial cells in the testis tissue of Paralichthys olivaceus.

The invention discloses application of a xanthoceras sorbifolia extract in preparation of a medicine for preventing and / or treating electromagnetic radiation damage. Experiments show that the xanthoceras sorbifolia bunge flower extract can improve the weight loss of rats caused by microwave radiation; brain tissue structure change damage caused by microwave radiation is improved; the reduction of the learning and memory ability of the rats caused by microwave radiation is improved; the brain sea horse tissue amino acid neurotransmitter disorder caused by microwave radiation is improved; electroencephalogram alpha and beta wave abnormity caused by microwave radiation is improved; the sperm motility reduction caused by microwave radiation is improved; the rat serum sex hormone disorder caused by microwave radiation is improved; rat testicular tissue oxidative stress injury caused by microwave radiation is improved; the abnormal energy metabolism of testicular tissues caused by microwave radiation is improved; heart electrophysiological abnormality and heart tissue ultrastructure abnormality caused by microwave radiation are improved; the immune function decline caused by microwave radiation is improved; the influence of microwave radiation on the content of IL-2, 4 is improved; the influence of microwave radiation on the thymus and spleen tissue structure is improved.

The invention discloses a method for synchronously marking germ cells and interstitial cells in paralichthys olivaceus testis tissues, which comprises the following steps: cloning paralichthys olivaceus MIR17HG-a (long non-coding RNA) by using a probe primer, and extracting plasmids to obtain target plasmids; the method comprises the following steps of: synthesizing a paralichthys olivaceus MIR17HG-a probe through SP6 / T7 RNA polymerase, preparing paralichthys olivaceus testis tissues into tissue slices, and performing fluorescence in-situ hybridization on the tissue slices by using the paralichthys olivaceus MIR17HG-a probe, so that the paralichthys olivaceus MIR17HG-a probe can synchronously mark germ cells and interstitial cells in the paralichthys olivaceus testis tissues. The defect that a traditional marking technology needs combined marking or multi-time step-by-step marking is overcome, and the blank of synchronous marking of germ cells and interstitial cells in paralichthys olivaceus testis tissue at present is filled.

The invention belongs to the technical field of culture media, and discloses a culture medium for in-vitro culture of cynomolgus monkey testicular tissue. Through an innovative culture technology and in combination with a culture formula of adding ANGPTL4 and IGF1, the survival rate and spermatogenesis efficiency of testis tissues are optimized, an efficient and reliable technical means is provided for reproductive medicine research and fertility preservation, and the method is particularly suitable for pre-adolescent individuals or wild protection animals or humans needing to preserve fertility potential. The innovative rotary culture technology and the culture scheme that 20 ng / mL of ANGPTL4 and 20 ng / mL of IGF1 are additionally supplemented are beneficial to the integrity of testicular tissues in the in-vitro culture process and promote proliferation and differentiation of germ cells, and the spermatogenesis efficiency is improved. And a feasible technical scheme and a new insight can be provided for the research of male animal in-vitro spermatogenesis.

The invention relates to the technical field of animal reproductive biology, and discloses a single-cell multi-omics method for analyzing a cattle testis development and spermatogenesis regulation mechanism, which comprises the following steps: (a) acquiring testis tissue samples of cattle at different development stages; (b) respectively carrying out single cell RNA sequencing (scRNA-seq) and single cell nucleus ATAC sequencing (snATAC-seq) on the testicular tissue sample, so as to obtain transcriptome data and chromatin accessibility data. According to the single-cell multi-omics method for analyzing the regulation and control mechanism of testis development and spermatogenesis of the cattle, by integrating single-cell RNA sequencing and single-cell nucleus ATAC sequencing, nine main cell types and specific marker genes (including newly found marker genes such as CPEB1, ADGRG1 and the like) of testis of the Simmental cattle from birth to puberty are systematically identified; the composition change rule of each cell type in different development stages is finely described, and a comprehensive cell map is provided for understanding the development of the bovine testis.

The invention belongs to the technical field of medicines, and particularly relates to application of a Chinese chive seed extract in preparation of a medicine for treating oligoasthenospermia. Based on network pharmacology prediction and animal experiment verification, it is found that by activating a PI3K / AKT signal channel, testicular cell apoptosis is inhibited, spermatogenesis disorder is improved, and therefore the sperm number and vitality of an oligoasthenospermia model rat are remarkably improved. Clinical treatment research in combination with levocarnitine oral liquid proves that the traditional Chinese medicine composition can reduce the sperm DFI level and improve the semen quality. Animal research proves that the compound can regulate the level of serum testosterone (T), reduce the levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH), repair pathological damage of testis tissues, and antagonize testis cell apoptosis induced by tripterygium glycosides by up-regulating protein expression of p-PI3K / PI3K, p-AKT / AKT and Bcl-2.

The invention discloses an RNA (Ribonucleic Acid) molecule, a nano-liposome and application of the RNA molecule to preparation of a product for treating testis injury and / or testis aging. The nucleotide sequence of the RNA molecule is as shown in SEQ ID NO. 1. The RNA molecule and the nano-liposome loaded with the RNA molecule provided by the invention show an excellent anti-inflammatory function and a remarkable targeting property in vivo, and most of the RNA molecule and the nano-liposome loaded with the RNA molecule can be taken by testis macrophages, so that the treatment effect is effectively improved, the repair and regeneration of testis tissues are promoted, the spermatogenic ability of testis is remarkably improved, the normal reproductive endocrine environment is recovered, and the curative effect is good. The testosterone level is improved. The invention provides a brand new technical strategy for treating testis injury, improving testis aging and improving male fertility, and is expected to play an important role in future clinical application.

The invention belongs to the technical field of biology, and provides a vitrification rapid cryoprotectant for preserving pre-puberty male testis tissue and a freezing method aiming at the problems that an existing testis tissue preserving method is unstable in preserving effect, long in consumed time and complex in operation. The cryoprotectant comprises a cryoprotectant A, a cryoprotectant B and a cryoprotectant C, the cryoprotectant A comprises a DMEM (Dulbecco Modified Eagle Medium), fetal calf serum and ethylene glycol; the cryoprotectant B comprises a DMEM (Dulbecco Modified Eagle Medium), fetal calf serum and ethylene glycol; the cryoprotectant C comprises a DMEM (Dulbecco Modified Eagle Medium), fetal calf serum, cane sugar and ethylene glycol. According to the method, the pre-puberty male testicular tissue can be effectively preserved at low temperature, the high cell viability is obtained, and the structural integrity of the testicular tissue is well preserved.

The invention discloses application of sodium 4-phenylbutyrate in a product for improving dyszoospermia of obese males, and belongs to the technical field of biological medicines. High-fat feed is adopted to construct a rat obesity model, the protection effect of 4-PBA on the reproductive function of obese rats is studied, and experimental results show that after the 4-PBA intervenes the obese rats, the testosterone content can be obviously increased, the estradiol level can be reduced, the sperm motility can be improved, the sperm malformation rate can be reduced, the spermatogenic dysfunction can be obviously relieved, and the anti-obesity effect on testis tissue morphology, testis tissue morphology, testis tissue morphology, testis tissue morphology and testis tissue morphology can be improved. The functions of supporting cell mitochondria, endoplasmic reticulum and MAM structures are obviously improved, and a new treatment strategy and a new drug target are provided for intervening and controlling obesity induced testis dyszoospermia and improving fertility of obese males.

The application discloses a pharmaceutical-grade PDRN and a preparation method thereof, and belongs to the technical field of biotechnology. The method comprises the following steps: (1) grinding salmon testis tissue in a pre-cooled grinding buffer to obtain a homogenate; (2) using Forskolin to induce the grinding homogenate; the induction conditions are as follows: first, adding Forskolin to 8-12 muM, and standing for 1-3 hours at 4 DEG C; then, increasing the temperature to 10-20 DEG C, adding Forskolin to 18-22 muM and 2 mM glutathione, and treating for 2 hours; and (3) extracting the PDRN through SDS-sodium chloride.

The application relates to a kit for detecting the differentiation state of fruit fly testis spermatogonia and an application thereof, wherein the kit comprises reagents for detecting the expression level of a Bckdhb gene, the Bckdhb gene being a dominant expression gene in fruit fly testis tissue. By using a UAS / Gal4 system to specifically knock down Bckdhb in spermatogonia and driving the Bckdhb by Bam-Gal4, it is found that the down-regulation of the Bckdhb expression level can cause the spermatogonia differentiation to be blocked, the testis top nucleus deepening distance is increased, the germ cell differentiation is defective, meanwhile, the Bckdhb mRNA level is significantly decreased, and the Piwi mRNA level is significantly increased. The expression level of the Bckdhb can be detected to effectively judge the differentiation state of the spermatogonia. The kit provided by the application can be used for fruit fly testis spermatogonia differentiation research, and has important scientific research application value.

The invention provides a separation method and an identification method of bactrian camel testis sertoli cells, and belongs to the technical field of cell separation. The method comprises the following steps: mixing 0.08-0.12% of collagenase IV solution and 0.23-0.27% of trypsin-EDTA (Ethylene Diamine Tetraacetic Acid) according to a volume ratio of 1: 1, digesting a bactrian camel testis tissue block, and then combining differential adhesion and hypotonic treatment to obtain bactrian camel testis supporting cells with the purity of 95% or above, meanwhile, reducing cell damage, and enabling the cell survival rate to exceed 85%; and the identification is completed by combining morphological observation, immunofluorescence staining and secretion function detection. The separated bactrian camel testicular sertoli cells can be used for double-hump reproductive performance research, and a foundation is laid for the reproductive performance of bactrian camels.

The invention provides a mononuclear transcriptomics technology-based analysis method for interaction of testicular sertoli cells and germ cells of plateau livestock, and belongs to the technical field of bioinformatics. The analysis method comprises the following steps: carrying out mononuclear transcriptome sequencing by taking testis tissues of plateau livestock at different development stages as materials, dividing a supporting cell subset and a germ cell subset according to marker genes, drawing a quasi-time differentiation track, screening differential expression genes and enriching functions, and constructing a supporting cell and germ cell interaction molecular network. According to the method, molecular characteristics, differentiation tracks and interaction networks of supporting cells and germ cells of the testis of the Tibetan sheep in different development stages are analyzed, and the problems that in the prior art, cell resolution is insufficient, development dynamics are deficient, variety specificity information is deficient and the like are solved; a brand new technical approach and theoretical basis are provided for clarification of cell molecular basis of low reproductive capacity of Tibetan sheep and mining of key regulatory factors.

The application belongs to the field of animal reproductive biotechnology, and discloses application of support cell exosomes in promoting in-vitro culture of pig testis tissue, the in-vitro culture comprising: placing pig testis tissue pieces on a three-dimensional culture support and culturing by using a sequential medium system; wherein the sequential medium system comprises M1 medium used for initial culture and M2 medium used for subsequent culture, the M1 medium comprising StemPro-34 SFM basic medium, and the M2 medium comprising alpha-MEM basic medium. The technical problems of poor in-vitro culture effect of testis tissue and difficulty in maintaining the spermatogenesis process in the prior art are solved.

The invention discloses a function and an application of virus source tyrosine phosphatase in development of male testis of plutella xylostella, namely an application of a protein tyrosine phosphatase gene CvBV22-9 coded by cotesia vestalis virus in induction of apoptosis of testis cells of larvae of plutella xylostella. According to the invention, the CvBV22-9 gene is screened and identified as a main factor for inducing testis apoptosis, and the gene is continuously and highly expressed in a parasitized testis tissue and has tissue specificity. After the expression of the CvBV22-9 is knocked down, the testis apoptosis rate is obviously reduced; and about 90.80% of testis cell apoptosis can be induced by using recombinant baculovirus to over-express the gene. The low-concentration recombinant baculovirus can also destroy the testis structure and lead to decrease of the egg laying amount of mating female insects. According to the invention, a transgenic fruit fly model is successfully constructed, specific expression of CvBV22-9 in fruit fly testis is realized by using a UAS-Gal4 system, and 97.30% of testis apoptosis is induced.

The invention belongs to the technical field of animal reproductive biology, and discloses an application of a sustentacular cell exosome in promoting in-vitro culture of a pig testis tissue, and the in-vitro culture comprises the following steps: placing a pig testis tissue block on a three-dimensional culture bracket, and culturing by adopting a sequential culture medium system; wherein the sequential culture medium system comprises an M1 culture medium for initial culture and an M2 culture medium which is used for subsequent culture and added with exosomes separated and extracted from porcine supporting cells, the M1 culture medium comprises a StemPro-34 SFM basal culture medium, and the M2 culture medium comprises an alpha-MEM basal culture medium. The technical problems that in the prior art, the testicular tissue in-vitro culture effect is poor, and the spermatogenesis process is difficult to maintain are solved.

The invention discloses an application of L-phenylalanyl-L-lysine in improving a spermatogenic function. The L-phenylalanyl-L-lysine is proposed for the first time to be capable of remarkably increasing sperm concentration, improving sperm malformation, improving sperm motility and effectively improving the spermatogenic function. Meanwhile, L-phenylalanyl-L-lysine is used for treating oligoasthenozoospermia, can promote thickening of spermatogenic tubule spermatogenic cell layers, increase spermatogenic cell areas, improve sperm differentiation and formation processes, relieve spermatogenic dysfunction of testicular tissues and increase the total number of sperms, is remarkable in treatment effect and high in safety, and has excellent clinical transformation prospects.

The application belongs to the technical field of animal models, and provides a method for constructing a mouse testis tissue stress granule animal model, which comprises the following steps: after the mouse is anesthetized, the testis and epididymis of one side of the mouse are taken out; a sodium arsenite solution containing fast green dye is injected into the testis of the mouse through the entrance of the rete testis, and the concentration of the sodium arsenite solution is 30-50 mM; the wound of the mouse is sutured, the mouse is killed after the mouse recovers for 3 hours, and the testis tissue is separated for an immunofluorescence experiment; when obvious stress granules are formed, it is indicated that the model construction is successful. The application also provides an application of the method for constructing an animal model in identifying the generation of stress granules of spermatocytes and in identifying the generation of RNA binding protein SERBP1. The application provides a method for exploring the role of SERBP1 protein in the stress granules of the mouse testis tissue, a related mechanism and a possible molecular pathway, and provides a new theoretical basis for guiding the generation and recovery of cell stress granules under special conditions.

A method and composition for modulating male fertility are disclosed, centered on the targeted alteration of transmembrane channel-like (TMC) protein function, activity, or gene expression. Small molecules or pharmaceutically acceptable salts or solvates thereof are provided to modulate the activity or expression of TMC proteins, such as TMC5, in testis tissue. The compositions may include TMC5 channel blockers, dimerization inhibitors, TMC5-CIB1 interaction inhibitors, or small interfering RNAs targeting TMC5 mRNA, optionally formulated with a pharmaceutically acceptable carrier and capable of crossing the blood-testis barrier. Methods of diagnosing male infertility are also provided, comprising quantitating TMC5 expression or activity and comparing to a standard to identify infertility. These approaches enable reversible, non-hormonal regulation of spermatogenesis and fertility, offering new therapeutic strategies for male contraception and infertility treatment.

The invention relates to the technical field of non-targeted metabonomics, and discloses an application of pterostilbene in preparation of a medicine for preventing or treating dyszoospermia, and the medicine contains pterostilbene as an active component. In addition, the pterostilbene inhibits oxidative stress reaction by adjusting PI3K / AKT / mTOR signal channels and improves the pathological structure of testis tissues so as to prevent or treat cyclophosphamide-induced spermatogenesis dysfunction. According to the application, CTX is used for establishing a mouse dyszoospermia model, network pharmacological analysis and experimental verification are combined to discuss key action targets and pathways of PTS for preventing and treating dyszoospermia and a specific molecular mechanism, and meanwhile, metabonomics is combined to obtain the pterostilbene capable of preventing cyclophosphamide-induced male testicular dysfunction.