86 results about "Testicular tissue" patented technology

The present invention provides a cancer-associated epitope consisting of two polypeptides, wherein the first polypeptide is derived from cytokeratin K8, and the second polypeptide is derived from cytokeratin K18. The cancer-associated epitopes become exposed during malignant transformation, especially in colon, breast, ovarian, kidney, lung and testicular tissues. Exposure of the cancer-associated epitopes occurs by cleavage and removal of the N-terminal peptides of cytokeratins K8 and K18. The invention also provides a cancer-associated epitope up to 10M-1 in cancer tissue, which is 100 times higher than that of cytokeratin K8 / K18 complex in normal tissue. The present invention provides cancer-associated epitopes, binding entities, antibodies and methods for detecting and treating cancer using the epitopes, binding entities and antibodies.

The invention provides a serum pancreatic cancer exosome specific protein, which comprises a protein ADAM10, a protein CFHR1, a protein APOA4, a protein testicular tissue protein Li 70, a protein HEL-S-78p, a protein PGLYRP2, a protein IGFALS, a protein BCHE, a protein ANPEP, a protein LBP, a protein C9, a protein PZP and a protein ANXA11. The invention further provides a re-verification process of the serum pancreatic cancer exosome specific protein, which comprises the steps of S1, separating the exosome-free serum and the exosome in a serum sample; S2, preparing an exosome PBS solution by adopting the exosome separated in the step S1; S3, checking the ultra-micro structure of the exosome through a transmission electron microscope; and S4, detecting the expression of some maker proteinson the surface of the exosome by through Western blot analysis. The serum pancreatic cancer exosome specific protein of the invention can be used for preparing related product for pancreatic cancer diagnosis, the re-verification process of the serum pancreatic cancer exosome specific protein has the advantages of absolute specificity, high throughput, no interference and ability of recognizing macromolecular isomers and small molecules.

The invention belongs to the field of biological engineering and provides a method for separating human spermatogonial stem cells. The method comprises the following steps of: mechanically separating a testicular tissue; processing the testicular tissue into a semi-liquid state; adding digestive enzyme solution I into the liquid testicular tissue in the semi-liquid state; digesting the testiculartissue into a single convoluted tubule; adding digestive enzyme solution II in the convoluted tubule; digesting the convoluted tubule into a single cell; separating reproductive cells from supportingcells in the single cell; separating G protein-coupled receptor 125 (GPR125) positive spermatogonial stem cells from glial cell line-derived neurotrophic factor (GDNF) family receptor alpha 1 (GFRA1)positive spermatogonial stem cells by adopting an immunomagnetic bead cell sorting method; and identifying the separated GPR125 positive spermatogonial stem cells and the GFRA1 positive spermatogonial stem cells by adopting an immune cell chemical method. By the method, a large number of the high-purity human spermatogonial stem cells are obtained, and sufficient cell sources are provided for theresearch of the human spermatogonial stem cells and the application of the human spermatogonial stem cells in regenerative medicine and reproductive medicine.

A method and device for freeze-drying a biological sample of mammalian cells or tissue are disclosed. The method includes placing a biological sample on or in a structure to increase a temperature ofthe biological sample and with the biological sample in a closed chamber applying a vacuum to the chamber to lower a pressure within the chamber, cooling the chamber to lower a temperature within thechamber and applying heat to the biological sample within the chamber. The biological sample can include one or more of stem cells, hematopoietic stem cells, mesenchymal stem cells, embryonic stem cells, induced pluripotent stem cells, sperm, oocytes, embryos, ovarian tissue, uterine tissue or testicular tissue. A method for rehydration of a sample is further disclosed.

The invention relates to a sperm freezing fluid and a preparation method thereof and belongs to the technical field of human assisted reproduction. The sperm freezing fluid is prepared by dissolving glycerinum into a salt solution with cane sugar, antibiotics and an indication agent in a concentration of 20-30%. Due to addition of the glycerinum and the cane sugar in the sperm freezing fluid provided by the invention, the glycerinum and the cane sugar are adopted as cell protection agents which can not only gently and effectively protect activity of sperms and completeness of DNA (Deoxyribonucleic Acid), but also protect the activity of testicular tissue cells, furthermore the activity of sperms in testicular testes is protected, the fertility rate of the sperms is increased, and the security of assisted reproductive technologies is powerfully ensured.

The invention discloses a method for two-dimensional electrophoresis extraction and separation of total protein of a rat and mouse testicular tissue sample. The method comprises rat and mouse testicular tissue treatment, a preparation method for a testicular tissue total protein sample, a two-dimensional electrophoresis technology system, gel dyeing and spectrum analysis. The method optimizes a tedious universal two-dimensional electrophoresis technology system with relatively poor stability, greatly reduces time and reagent consumption for finding out conditions, saves experiment cost, and obtains a satisfactory two-dimensional electrophoresis spectrum that is convenient for subsequent analysis. The method is more suitable for the extraction of the rat and mouse testicular tissue protein sample than a universal homogenate extraction method, a lysate extraction method, a lysate extraction / acetone precipitation method at present. The method not only reduces the degradation of the protein but also increases solubility of the protein without increasing cost of the reagent, so that the two-dimensional electrophoresis spectrum with relatively many protein sites, clear horizontal stripes and relatively good repeatability can be obtained.

The invention relates to a method for separating pig spermatogonia stem cells. The method comprises steps of preparing solutions and separating spermatogonia stem cells, wherein in the step of preparing solutions, the solutions to be prepared comprises nutrient solution, digestive enzyme solution, buffer solution, collagen solution, gelatin solution and laminin solution, the step of separating spermatogonia stem cells comprises substeps of cutting pig testicular tissue into pieces and adding enzyme for digestion, filtering by a cell strainer, carrying out enrichment culture and difference adherent culture of the spermatogonia stem cells, finally collecting the spermatogonia stem cells, wherein the difference adherent culture comprises collagen coating adherent culture and laminin coating adherent culture. The method is characterized in that the nutrient solution comprises DMEM / F12, FBS (fetal bovine serum), sodium pyruvate, penicillin and streptomycin; according to the collagen coating adherent culture, after being cultured for required time, the cell is blown and beaten and then continuously cultured. In the collagen coating adherent culture, the spermatogonia stem cell is slightly blown and beaten and is not attached to the wall of the collagen, so that loss of the spermatogonia stem cell is reduced, and the yield of the spermatogonia stem cell is improved.

The invention discloses application of astragalin in preparation of a drug for treating diabetic testicular dysfunction. An experimental result shows that when the astragalin is used in a safety doserange, the dose is 30 mg / kg of astragalin, the astragalin can remarkably protect sperm injury induced by a diabetes model, improve pathologic change of testicular tissue, improve the oxidation resistance ability and regulate and control protein expression of inflammatory cytokines, and it is proved that the astragalin has the effect of improving the diabetic testicular dysfunction.

The invention discloses a complete testis culture medium and application thereof. The method comprises the following steps: separating human testis tissues into small blocks, placing the small blocks on 0.25-1.5% of agar blocks which are subjected to complete testis culture medium replacement for at least 8 hours in advance, adding a complete testis culture medium to reach 1 / 3-2 / 3 of the height of the agar blocks, and culturing for 10-100 days in a 5% carbon dioxide environment at 32-37 DEG C. The model provides a platform for in-vitro research on human testicular spermatogenesis, and provides a research platform for supporting research aiming at understanding testicular organ generation complexity and developing urgently needed male infertility treatment.

The invention discloses a method for separating and culturing pig spermatogonial stem cells by a mechanical process. The method of the invention is characterized in that the characteristics of animal testicular tissue are combined, and the testicular tissue can be separated by the mechanical process and then cultured, so that the method for separating and purifying spermatogonial stem cells can be enriched. According to the invention, the pig spermatogonial stem cells can be obtained without enzyme digestion, the method of the invention has the advantages of fast cells clone and good state ofcells, and solves the problems of difficult separation, small quantity and high requirement of culture condition of spermatogonial stem cells. Compared with the separating technology of the prior art, the method of the invention has simple operation, and is capable of reducing the difficulty to obtain cells, substantially reducing the experimental cost and raising the success rate of the experiments.

The invention relates to a cell nucleus size and shape measuring method. Testicular tissue slices of male animals are analyzed by using a microscopic image analysis system, so that leptotene stage spermatocytes, pachytene stage spermatocytes and spermatids are quickly and automatically classified. The method has huge advantages of quickness, accuracy and human and material resource saving, and provides an important evaluation basis for reproductive system toxicity evaluation of dogs and other male animals in non-clinical safety evaluation.

The invention discloses an application of dendrobium polysaccharide in preparation of drugs to prevent or restore reproductive injury after chemotherapy. The dendrobium polysaccharide applied to prevent and treat male reproductive injury caused by radiotherapy and chemotherapy has obvious therapeutic effect and small side effects, and can effectively reduce the damage caused by nitrogen mustard anticancer drugs to the reproductive system, which not only makes the sperm quantity of mice rise significantly, but also improves the sperm quality, and effectively improves the antioxidant activity ofmouse testicular tissue, and in mouse testicular tissue, the level of SOD, GPx, GSH and CAT is obviously increased while the level of MDA is obviously decreased. The dendrobium polysaccharide with wide source of raw materials is simple in extraction and purification method, high in sugar content and suitable for large-scale production and popularization and has a good prospect in the preparationand / or treatment of reproductive injury after chemotherapy.

The invention provides a cryopreserved recovery reagent set for a pet testicular tissue. The cryopreserved recovery reagent set comprises (1) a cryopreserved reagent 1: 2-7W / V% of lycium barbarum polysaccharide, 0.5-5W / V% of cholesterol, 6-14V / V% of dimethyl sulfoxide, 2-7V / V% of propylene glycol and 80-90V / V% of MEM culture medium; (2) a cryopreserved reagent 2: 8-10W / V% of lycium barbarum polysaccharide, 0.5-5W / V% of cholesterol, 6-20V / V% of dimethyl sulfoxide, 5-15V / V% of propylene glycol and 70-80V / V% of MEM culture medium; (3) a cryopreserved reagent 3: 15-20W / V% of lycium barbarum polysaccharide, 0.5-5W / V% of cholesterol, 6-20V / V% of dimethyl sulfoxide, 5-15V / V% of propylene glycol and 70-80V / V% of MEM culture medium. The invention also provides a recovery reagent set for the pet testicular tissue and a cryopreserved recovery method for the pet testicular tissue.

The invention discloses a colloidal gold reagent strip for rapidly detecting Boule protein. A sample pad, a gold label pad, a chromatography film and a water absorption pad of which the ends are sequentially connected with one another are arranged on a rubber plate according to the sample flow direction; a detection area T and a quality control area C are formed in the chromatography film; the sample refers to supernatant of lysate of testicular tissue to be detected; a colloidal gold-anti-Boule protein polyclonal antibody-1 conjugate is adsorbed to the gold label pad; an anti-Boule protein polyclonal antibody-2 is coated in the detection area T; an anti-rabbit IgG is coated in the quality control area C. The detection method comprises the following steps: 1, taking the supernatant of lysate of testicular tissue to be detected; 2, dripping 2-3 droplets of the supernatant of lysate of testicular tissue to be detected on the sample pad; and 3, observing the result within 5 minutes. According to the reagent strip, the existence of the Boule protein in the lysate of the testicular tissue is detected by adopting a double antibody sandwich method and an immunochromatographic method. The detection process is high in sensitivity, high in specificity and high in accuracy, the sperm development block condition is clear at a time, the detection speed is high, the result can be obtained within 5 minutes, and the operation is simple.

The invention discloses combined enzyme for efficiently digesting goat testicular tissue. The combined enzyme comprises collagenase, deoxyribonuclease, neutral protease, and ethylene diamine tetraacetic acid. The combined enzyme has the advantages of simple formula and good usage effect, and can obtain single cell from the goat testicular tissue in a stable mode, the cell survival rate and proportion of germline stem cell in the obtained cell are high, and the after culture, the typical germline stem cell clone appears.

The invention discloses a separation and culture method of human testicular mesenchymal stem cells. The invention researches and provides a specific marker, namely CD248 protein, specifically expressed in the human testicular mesenchymal stem cells (SLC) in human testicular tissues so as to identify and separate high-purity human SLC from the human testicular tissues. Therefore, a method for separating and obtaining the high-purity SLC from the human testicular tissues is established; and the obtained SLC does not have other types of cells in a testis and has high purity. Meanwhile, a culturesystem of the human SLC is optimized and improved; a serum-free culture medium of the obtained testicular mesenchymal stem cells has clear components and is good for standardization; and an efficientand stable culture and amplification system of the testicular mesenchymal stem cells is established. According to the separation and culture method disclosed by the invention, the high-purity human SLC can be obtained from the human testicular tissues and high-quality seed cells are provided for clinical transformation of treating male hypogonadism by the testicular mesenchymal stem cells; and theseparation and culture method has important meanings on promoting characteristic researches of the human SLC, especially clinical transformation of the SLC.

The invention discloses an application of dendrobium polysaccharide in the preparation of reproductive injury recovery drugs after cancer chemotherapy. The dendrobium polysaccharide has obvious therapeutic effect and small side effects, which can effectively reduce the damage caused by platinum anticancer drugs to the reproductive system, not only the sperm quantity of mice is significantly increased, the sperm quality is also improved, the antioxidant activity of mouse testicular tissue is also effectively improved, and in mouse testicular tissue, the level of SOD, GPx, GSH and CAT is obviously increased while the level of MDA is obviously decreased. The dendrobium polysaccharide with wide source is simple in extraction and purification method, and the obtained dendrobium polysaccharide is high in sugar content and suitable for large-scale production; the dendrobium polysaccharide has a good prospect in the preparation and clinical application of reproductive injury recovery drugs after cancer chemotherapy.

The invention discloses an application of bioactive polypeptide PACAP in preparation of drugs for improving fertility of obese men, and belongs to the field of damaged fertility diseases of the obesemen. The bioactive polypeptide PACAP drugs provided by the invention can significantly prevent and relieve obesity-induced male fertility damage (mainly by improving blood lipid and sex hormone levels, reducing a testicular tissue oxidative stress level, improving obesity-induced testicular spermatogenic cell damage, inhibiting high lipid-induced apoptosis of spermatogonia GC-1, and improving sperm function and early embryo quality after in-vitro fertilization), and repair pathological change and reproductive system function of fertility damage of the obese men, the relationship between obesity and male infertility can be initially understood, the protective effect of the PACAP on obesity-induced male reproductive damage can be clarified, and a theoretical basis, an experimental basis anda targeted treatment strategy are provided for clinical diagnosis and treatment of obese male infertility.

The invention discloses a human testicular tissue suspension cryoprotectant and a preparation method thereof. The human testicular tissue suspension cryoprotectant comprises the following raw materials: sodium chloride, potassium chloride, calcium chloride, magnesium chloride, monopotassium phosphate, sodium bicarbonate, HEPES, caffeine, sodium pyruvate, pentoxifylline, glycine, glucose and sucrose. The human testicular tissue suspension cryoprotectant also comprises sodium lactate and glycerol. The invention further discloses the preparation method of the human testicular tissue suspension cryoprotectant and a freezing method adopting the human testicular tissue suspension cryoprotectant. The cryoprotectant is specially designed for a testicular tissue suspension, the testicular sperm cryopreservation and resuscitation effect can be improved, the problems that potential safety hazards exist, the cryopreservation and resuscitation effect is poor, and movable testicular sperms are not easy to find are effectively solved, and the human testicular tissue suspension cryoprotectant has quite important clinical application value.

The invention provides application of luteolin to the preparation of a medicine for treating an oxidative stress injury of testicular tissue, and relates to the field of male reproduction and endocrinic medicines. An active component of a medicinal composition is the luteolin. Discovered through an experiment, the injury, which is caused by hydrogen peroxide and triptolide, of a testicular sustentacular cell can be reversed through the intervention treatment of the luteolin; the apoptosis of the testicular sustentacular cell induced by the hydrogen peroxide is inhibited; the accumulation of ROS (Reactive Oxygen Species) is inhibited; the collapse of a cellular mitochondrial membrane potential caused by the hydrogen peroxide is reversed; the expression of a downstream antioxidase gene can be activated; the autologous oxidation-resistance capacity of the sustentacular cell is improved; the injury to the testicular tissue of a mouse induced by the triptolide is effectively reversed. Besides, the luteolin can be also used for inhibiting the increment of the permeability of a blood-testis barrier, which is caused by the triptolide-induced oxidative stress injury, through regulating and controlling gap junction, occlusion junction and tight junction protein; the completeness of the blood-testis barrier is protected. Therefore, the luteolin can be applied to the preparation of the medicine for treating the oxidative stress injury of the testicular tissue.

The present invention provides an active substance of Morchella, its use and a composition thereof for improving disturbance of reproductive function, especially, for manufacturing a pharmaceutical composition to improve disturbance of reproductive function induced by obesity or metabolic syndrome. The composition with the active substance of Morchella can effectively improve the structural integrity of testicular tissues and sperm, increase testosterone levels in the blood, and reduce the oxidation stress in sperms.

The invention belongs to the field of biological engineering and provides a method for separating human spermatogonial stem cells. The method comprises the following steps of: mechanically separating a testicular tissue; processing the testicular tissue into a semi-liquid state; adding digestive enzyme solution I into the liquid testicular tissue in the semi-liquid state; digesting the testicular tissue into a single convoluted tubule; adding digestive enzyme solution II in the convoluted tubule; digesting the convoluted tubule into a single cell; separating reproductive cells from supporting cells in the single cell; separating G protein-coupled receptor 125 (GPR125) positive spermatogonial stem cells from glial cell line-derived neurotrophic factor (GDNF) family receptor alpha 1 (GFRA1) positive spermatogonial stem cells by adopting an immunomagnetic bead cell sorting method; and identifying the separated GPR125 positive spermatogonial stem cells and the GFRA1 positive spermatogonial stem cells by adopting an immune cell chemical method. By the method, a large number of the high-purity human spermatogonial stem cells are obtained, and sufficient cell sources are provided for the research of the human spermatogonial stem cells and the application of the human spermatogonial stem cells in regenerative medicine and reproductive medicine.

The invention discloses circRNA of target competitive binding of oar-miR-29b and an application of the circRNA. Healthy male Tibetan sheep testicular tissues in different development stages are used as test materials, circular RNA in testis is identified and analyzed through an RNA-seq technology and a related bioinformatics analysis method so as to dig the circRNA related to testis development and function maintenance, the circRNA is circ_024949 and / or circular RNA circ_026259, the nucleotide sequence of the circRNA circ_024949 is shown as SEQ ID No:1, and the nucleotide sequence of the circular RNA circ_026259 is shown as SEQ ID No:2. On the basis, the invention discloses the application of the circRNA as a marker for detecting the testis function or development condition of Tibetan sheep, to preparation of a medicine for regulating and controlling the testis function or development of the Tibetan sheep and to expression analysis or research of a downstream target gene BOLL. The circular RNA circ_029155 can be used for detecting and evaluating the testis development and spermatogenesis functions of the sheep, and lays a foundation for further exploring a molecular action mechanism of the testis development and spermatogenesis of the sheep and subsequent application and development.

The invention provides an application of dendrobium polysaccharide in the preparation of drugs for prevention and treatment of reproductive injury caused by arsenic poisoning. The dendrobium polysaccharide has obvious therapeutic effect on reproductive injury induced by arsenic exposure in male rats, which can significantly increase the number and motility of sperms, increase the incidence of sperm acrosome reaction and decrease the rate of sperm deformity; the SOD activity, GPx activity, CAT activity and GSH content in testicular tissue are also significantly increased, while MDA content is also significantly decreased; the dendrobium polysaccharide has rarely toxic and side effects and no adverse effect on normal growth and physiological state. The dendrobium polysaccharide with wide source of raw materials is simple in extraction and purification method, high in sugar content and suitable for large-scale production and promotion and has a good prospect in the preparation and / or treatment of reproductive injury drugs caused by arsenic poisoning and clinical application.

The invention discloses a method for constructing a machin progenitor leydig cell immortal line. The method comprises the following steps: by taking machin testicular tissues, obtaining leydig cells of a high ratio, culturing in a complete medium for 20-28 hours, replacing fresh culture solution, continuously culturing, and performing cell passage to the leydig cells presenting fibroblast growth in uniform form; infecting the leydig cells by utilizing slow virus solution carrying Large T and Egfp, performing fluorescence labeling cell screening, and performing cell passage to a cell strain for stably expressing Egfp; and screening the cell strain for stably expressing Egfp, identifying the progenitor leydig cells, continuously performing cell passage for over 50 generations, and finally constructing the machin progenitor leydig cell immortal line. The in-vitro cell passage of the progenitor leydig cell immortal line with the transferred Large T exceeds 62 generations, the leydig cell labeled proteins such as CYP11A1, CYP17A1 and STAR are expressed, marker genes of mesenchymal stem cells such as Pdgfral and Lifr are expressed, the cell shape is maintained into the fiber shape, and a characteristic progenitor leydig cell immortal line is obtained.

The invention provides a novel application of plant fruit pulp. The invention relates to application of plant fruit pulp in testis function protection, application of plant fruit pulp in prevention and / or treatment of oligospermia and asthenospermia, application of plant fruit pulp in protection of male fertility and application of plant fruit pulp in prevention and / or treatment of testis tissue lesion, testosterone level decline, testis function decline, sperm number reduction, asthenospermia and asthenospermia caused by metabolic disorder. The sperm motility rate is reduced; the sperm morphology is abnormal. The plant fruit pulp can effectively prevent and treat symptoms such as testicular tissue lesion, testosterone level decline, testicular hypofunction, sperm number reduction, sperm motility rate reduction and sperm morphology abnormality caused by metabolic disorder. The application provides a basis for protecting testis functions, preventing and / or treating oligospermia and asthenospermia and protecting male fertility by directly utilizing the plant fruit pulp.

The invention provides a molecular breeding method based on the polymorphism of a candidate gene PRL (Prolactin) and relates to the technical field of poultry breading, in particular to a method for utilizing the polymorphism of the candidate gene PRL to perform molecular breeding. Under the condition of focusing on a gene effect of a PRL gene in a gonadal axis of a chicken, i.e. hypothalamus-hypophysis-ovary or testis tissue to reproductive characters of the chicken, mRNA differential expression or difference of translating proteins thereof, factors influencing the expression of the reproductive characters of the chicken are researched and a genetic molecular mechanism of the reproductive characters of the chicken is primarily illustrated, so that a function of the candidate gene during a reproductive developing process can be objectively understood, and meanwhile, the analysis to the influence thereof on a reproductive performance can be facilitated, a basis for marking auxiliary breeding can be provided, the improvement of the reproductive characters can be quickened and the potential economic benefit of the candidate gene can be utilized.

A method for freeze-drying a biological sample of mammalian cells or tissue including placing a biological sample on or in a structure to increase a temperature of the biological sample and with the biological sample in a closed chamber applying a vacuum to the chamber to lower a pressure within the chamber, cooling the chamber to lower a temperature within the chamber and applying heat to the biological sample within the chamber. The biological sample can include one or more of stem cells, hematopoietic stem cells, mesenchymal stem cells, embryonic stem cells, induced pluripotent stem cells, sperm, oocytes, embryos, ovarian tissue, uterine tissue or testicular tissue.

The invention belongs to the technical field of instruments for reproductive medicine, and particularly relates to a package instrument capable of improving sperm-obtaining efficiency. The package instrument mainly comprises a fixing device and a poking device; the fixing device comprises a fixing rod, a first connection rod, a second connection rod and a fixing frame; the first connection rod isdetachably connected with the lower portion of the fixing rod, a second connection rod is connected with the lower portion of the first connection rod, and the second connection rod is connected withthe fixing frame; the poking device comprises a tissue poking rod, a third connection rod and a tissue poking device; and the third connection rod is detachably connected with the lower portion of thetissue poking rod, and the tissue poking device is connected with the lower portion of the third connection rod. The package instrument capable of improving the sperm-obtaining efficiency has the beneficial effects that by using the package instrument, testicular tissue can be well fixed, and damage to sperms in traditional sperm-obtaining operation processes and the like is avoided at the same time, so that the probability of success of sperm-obtaining operation is significantly increased.

The invention discloses novel applications of exogenous nerve growth factor (NGF). Establishment of rat cryptorchism models is realized; control groups are obtained via exogenous injection of NGF; real-time quantitative PCR, in-situ hybridization, immunofluorescence, immunohistochemistry, and transmission electron microscopy are adopted so as to observe morphological change of cryptorchidism tissue, and expression change of NGF, HoxA10mRNA and protein in testicular tissue, and confirm that NGF is capable of promoting formation of testicular sperms. So that, NGF can be widely used for preparation of medicines which are used for promoting sperm generation, promoting cryptorchism model testicular tissue development, reducing cryptorchidism tissue damage, and treating cryptorchidism.