35 results about "Seminiferous tubule structure" patented technology

The invention discloses novel growth hormone releasing hormone analog peptides and an application thereof in preparing medicines for treating infertility. Experiments discover that 2D, 2E or 2F peptide has obviously high hypophysis GH releasing activity and hypophysis hormone releasing specificity. Tested by the conjugation reaction of an in-vitro GHRH dimer peptide and a hypophysis GHRH receptor, 2D, 2E and 2F dimer peptides have extremely high hypophysis receptor binding activity, wherein the 2F dimer peptide has the maximum binding activity. With the 2F peptide as the representative, infertility model treatment finds out that, compared with a normal saline group and a pure cyclophosphamide control group, in the 2F dimer peptide group, the spermatocytes and the spermatogonia in the seminiferous tubules are obviously increased, the seminiferous tubule cells are arranged in order and large in volume, the cavities of the seminiferous tubules are reduced and even disappear, and dose dependency is shown. All the facts indicate that the GHRH peptides with the 2F peptide as the representative have an obvious effect of stimulating the proliferation and the maturation of spermatogonia/oogonia, thereby promoting reproduction; and as a result, the novel growth hormone releasing hormone analog peptides can be applied to medicines for treating infertility.

The invention discloses a separation and culture method and application of testicle mesenchymal stem cells. The method comprises the following steps: separating a testicle from a 7-day or 2-month mouse, removing the envelope and vessels to expose a seminiferous tubule, digesting the seminiferous tubule with collagenase IV, and filtering the digested cell suspension; and on the basis of the cell specific marker of CD51, separating with a flow cytometry to obtain single cells, and collecting the cells for expressing CD51, thereby separating the CD51 positive testicle mesenchymal stem cells. The invention also discloses testicle mesenchymal stem cells separated according to the method, self-renewal and proliferation cells with multi-directional differentiation potential obtained in a culture medium, and application of the cells in preparing compositions for treating diseases related to low testosterone level.

A low effective dose of a biologically acceptable chemical sterilant is injected into the dorsal cranial portion of a scrotal testis of a male animal for the purpose effecting sterilization of the animal. The injection of the chemical sterilant into the dorsal cranial portion has an effect on the epithelium of the tubuli recti, rete testis and ductus efferentes in addition to stopping spermatogenesis in the seminiferous tubules.

The invention discloses a separation and culture method of testicular mesenchymal stem cells for expressing nidogen and an application of the testicular mesenchymal stem cells. The separation and culture method comprises the following steps: providing a nidogen-green fluorescent protein transgenic mice model adopting C57BL/6 as the genetic background; raising the mice for one or more than one week, separating the testis, removing envelopes and blood vessels, exposing the seminiferous tubule, then digesting by using collagenase IV, and filtering digested cell suspension; and sorting by a flow cytometer to obtain single cells and collecting the cells for expressing green fluorescent protein with the fluorescence intensity being 10 times or more than 10 times of the fluorescence intensity of negative control cells so as to separate the testicular mesenchymal stem cells for expressing the nidogen. The invention provides testicular mesenchymal stem/progenitor cells separated according to the preparation and culture method and self-renewed and proliferated cells obtained by culture in a culture medium and provides the application of the cells in preparing a composition for treating related diseases with low testosterone level.

The invention relates to an application of an antioxidant drug in the pharmaceutical science field, and especially relates to an application of N-acetylcysteine in preparation of drugs for preventing and treating gonads and other tissue DNA damage and tissue damage induced by synchrotron radiation X-rays. According to experimental results, N-acetylcysteine is indicated to be capable of protecting the double-strand DNA damage caused by synchrotron radiation X-ray radiation, and protecting testicular seminiferous tubule cell layer number reduction caused by the synchrotron radiation X-ray radiation; and the antioxidant is indicated to have important application value in the preparation of the drugs for preventing and treating the synchrotron radiation damage.

The present invention relates the function of Dickkopf 3 (Dkk3 ) in testis using shRNA mediated knock down model. Specifically, the present invention provides knock down model comprising reduction in Dkk3 activity. The knock down model of Dkk3 consisting of non human vertebrates which have, incorporated in their genome, shRNA construct targeting mammalian Dkk3 gene exhibits low testis weight, low sperm count and has low litter size. The knock down model displays disrupted seminiferous tubules and are subfertile. The present invention model has a role in sex determination as its interruption leads to sex reversal of XY gonads, converting males to females. Sex reversal role of Dkk3 knock down model can find its utility in agricultural applications. The present invention describes Dkk3 as a dual function protein which is associated with sex determination as well as is essential for the process of spermatogenesis. Such testicular functional studies are useful as a model for various disease states of infertility or subfertility and for identifying a potential treatment to overcome idiopathic infertility.

The invention relates to application of LamG3/4/5 of a laminin alpha2 functional region in preparation of a medicine for preventing male contraceptive induced testis injury, and belongs to the technical field of medicine preparation. An amino acid sequence of the LamG3/4/5 of the laminin alpha2 functional region is shown as SEQ ID NO. 1. Overexpression drugs of LamG3/4/5 of the laminin alpha 2 functional region can effectively prevent testis injury induced by male contraceptives, and can play an obvious role in preventing structural integrity damage of seminiferous epithelium of seminiferous tubules in a testis, lesion of the seminiferous tubules and cytoskeletal protein damage/destruction of the seminiferous epithelium of the seminiferous tubules in the testis which are caused by oral administration of Adjudin.

The invention discloses a preparation method of a buffalo testis single-cell suspension solution. The preparation method comprises the following steps: (1) pre-treating raw materials: taking a buffalotestis and disinfecting, then shearing to remove tunica albuginea; washing and cutting into small blocks; (2) digesting: transferring the sheared testis into a culture dish; adding a primary digestive juice for digesting until single seminiferous tubules are obtained; centrifuging and removing supernatant; (3) carrying out primary re-suspension: re-suspending precipitates by utilizing a PBS (Phosphate Buffer Solution); centrifuging and removing supernatant; after repeating for two times, obtaining a bottom-layer re-suspension solution; (4) carrying out cell separation: after carrying out secondary digestion, placing the solution on a microscope and observing; blowing through a pipette to form single cells; after neutralizing, collecting the cells into a centrifugal tube and centrifuging;removing supernatant to obtain a primary cell suspension solution; (5) carrying out secondary re-suspension: re-suspending by utilizing a culture solution and filtering through a cell net to obtain the buffalo testis single-cell suspension solution. The invention provides the preparation method of a buffalo spermatogonial stem cell suspension solution, which has high separation speed and good effect; the method is simple and feasible to operate and is easy to realize.

A method for detecting the nucleoproteins of spermatogenic cells through utilizing indirect immunofluorescent staining comprises the following steps: separating convoluted seminiferous tubules from the testis tissues of an animal, utilizing a hypotonic solution to process spermatogenic cells, and detecting the nucleoproteins of the spermatogenic cells through adopting indirect immunofluorescent staining, wherein the hypotonic solution comprises 17mM of sodium citrate, 50mM of sucrose, 5mM of EDTA, 0.5mM of DTT, 0.5mM of PMSF and 30mM of Tris and having a pH value of 8.2. The method uses the hypotonic solution to carry out permeable treatment of the cells in order to rapidly expand the cells, disperse chromatins and fully release the proteins in cell nuclei, so the spermatogenic cells can well combine with primary antibodies, thereby the combination efficiencies of secondary antibodies are enhanced, fluorescent staining is complete and full, and the preparation of spermatogenic cell chromosomes and karyotyping are successfully carried out.

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 relates to a testicular tissue cryopreservation method based on single seminiferous tubule perfusion, which comprises the following steps: making testicular tissue into the single seminiferous tubule, soaking a single seminiferous tubule in a cryopreservation protective agent 1 for 10 minutes, and soaking and perfusing the seminiferous tubule by using a cryopreservation protective agent 2, so as to maximally reduce the permeation damage of the protective agent while achieve vitrification cooling, quickly transferring the poured spermatogenic tubule to a Cryo-genic single sperm cryopreservation slice, after sleeving of the Cryo-genic tubule, directly conducting immersing in liquid nitrogen to quickly achieve cooling, and quickly conducting soaking in a recovery solution aftercooling. Compared with a traditional testicular cell suspension cryopreservation method, the method disclosed by the invention has the advantages that (1) the complete structure of the seminiferous tubule is reserved; (2) the oxidative stress level of testicular cells is effectively reduced; (3) the death of a large number of spermatogonial cells and supportive cells after cryopreservation is effectively reduced; and (4) the recovery rate and activity of sperms in the spermatogenic tubules are high.

The invention discloses use of cyanidin-3-O-glucoside (C3G) and/or protocatechuic acid (PCA) in improving heat tolerance of testis. The cyanidin-3-O-glucoside and/or protocatechuic acid can relieve phenomena of atrophy of testis seminiferous tubules, vacuolation of spermatogenic epithelium, reduction of diameter and thickness of testis lumens and the like. The C3G and/or the metabolite PCA thereof can reduce a phosphorylation level of eIF2alpha and an expression of stress particles by reducing an external stress pressure suffered by the testis so as to increase heat tolerance of testis, improve an antioxidant system of the testis and regulate an IRE1alpha-XBP1 pathway so as to relieve oxidative stress and endoplasmic reticulum stress-mediated apoptosis and relieve spermatogenesis disorder and testis injury induced by heat stress. The C3G and/or the metabolite PCA thereof provides a reliable theoretical basis and hasimportant significance for relieving a male reproductive injury induced by the testis in a heat stress state due to bad living habits, occupational factors and pathological reasons in the future.

The invention provides application of a sea grape extract in preparation of a pharmaceutical composition or health food. The sea grape extract is obtained by hot water extraction, can improve fasting blood-glucose, insulin concentration in blood and oral glucose tolerance so as to relieve diabetes mellitus, and can improve sperm motility, sperm variation and seminiferous tubule atrophy so as to prevent or treat reproductive disorder, wherein the application mode of the pharmaceutical composition comprises oral administration, soaking, spraying or injection, and the effective dose of the sea grape extract in the human body is 40-100 mg/kg per day.

A transgenic method using recombinant retrovirus as mediation to transfect stem spermatogonium in body includes cloning target gene to retrovirus carrier, transforming PA317 cells, collecting and concentrating the recombinant virus generated by clone of positive cells, injecting it in the contorted seminiferous tubule of mouse, and reproducing filial generation. Its advantages are simple operation and high repeatability.

The invention discloses an SCOS mouse modeling prejudging method based on Raman spectrum, comprising the following steps: a, obtaining mouse sample Raman spectrum feature; b, obtaining Raman spectrum seminiferous tubule spectral peak vector sample database based on the obtained mouse sample Raman spectrum feature; c, obtaining seminiferous tubule Raman spectrum based on the obtained Raman spectrum seminiferous tubule spectral peak vector sample database; d, extracting feature spectral peak based on the obtained seminiferous tubule Raman spectrum; and e, detecting the seminiferous tubule based on the extracted feature spectral peak. The SCOS mouse modeling prejudging method based on Raman spectrum, provided by the invention, can overcome the defects in the prior art, such as complex operation process, long consumed time and low reliability, so as to realize the advantages of simple operation process, short consumed time and high reliability.

The invention discloses an automatic segmentation method for various tissues in a mouse testis pathological section based on deep learning. The method belongs to the field of machine learning and image processing. The method comprises the following specific steps: pretreating mouse testis cross section slices; establishing a mouse seminiferous tubule segmentation model based on ResNet; and establishing Unet-based segmentation of multiple types of germ cells and multiple types of tissue regions in the mouse seminiferous tubule. Firstly, seminiferous tubule pre-segmentation performedon a mouse testis cross section full-scanning image by using ResNet in combination with a sliding window and a pixel-by-pixel segmentation method; then, multiple types of cell nucleuses and multiple types of tissue regions in the seminiferous tubule are respectively segmented by using Unet. According to the method, good performance is obtained, and a good image analysis basis is provided for establishing a mouse seminiferous tubule automatic staging system; in the future, technicians extract histological characteristics of cell nucleuses and tissue regions in the mouse seminiferous tubule, and the histological characteristics are used for training an automatic staging classifier of the mouse seminiferous tubule.