[go: up one dir, main page]

WO2019161707A1 - Procédé d'établissement d'un modèle simiesque de déficience en testostérone - Google Patents

Procédé d'établissement d'un modèle simiesque de déficience en testostérone Download PDF

Info

Publication number
WO2019161707A1
WO2019161707A1 PCT/CN2018/123563 CN2018123563W WO2019161707A1 WO 2019161707 A1 WO2019161707 A1 WO 2019161707A1 CN 2018123563 W CN2018123563 W CN 2018123563W WO 2019161707 A1 WO2019161707 A1 WO 2019161707A1
Authority
WO
WIPO (PCT)
Prior art keywords
eds
testosterone
injection
testicular
injected
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2018/123563
Other languages
English (en)
Chinese (zh)
Inventor
项鹏
柯琼
夏凯
邓春华
毛富祥
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sun Yat Sen University
Original Assignee
Sun Yat Sen University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sun Yat Sen University filed Critical Sun Yat Sen University
Publication of WO2019161707A1 publication Critical patent/WO2019161707A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/095—Sulfur, selenium, or tellurium compounds, e.g. thiols
    • A61K31/10—Sulfides; Sulfoxides; Sulfones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/255—Esters, e.g. nitroglycerine, selenocyanates of sulfoxy acids or sulfur analogues thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2207/00—Modified animals
    • A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/106—Primate
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0306—Animal model for genetic diseases

Definitions

  • the invention belongs to the field of animal models, and in particular relates to a method for establishing a testosterone deficiency model of monkeys.
  • Testosterone deficiency syndrome refers to the abnormality of the target organ morphology and function caused by insufficient testosterone levels in various stages of men's life, which leads to the corresponding clinical symptoms. It is closely related to the occurrence and development of major diseases such as fertility, sexual dysfunction, osteoporosis, depression, cardiovascular and cerebrovascular diseases, diabetes and metabolic syndrome. It is a male reproductive endocrine disease that seriously affects men's health, life expectancy and quality of life. TDS mainly includes late onset hypogonadism (late-onset hypogonadisim (LOH)), congenital hypogonadism, testicular injury and other diseases.
  • LH late onset hypogonadism
  • congenital hypogonadism congenital hypogonadism
  • TDS mainly uses exogenous testosterone drug supplementation therapy, supplementation of exogenous testosterone by oral or injection, although there is a certain effect, but due to obvious defects such as inconvenient use and side effects, only very A small percentage of patients with TDS receive this therapy. Therefore, exploring a new method for treating TDS has become a key scientific and technological problem that urgently needs to be solved in the social development of our country.
  • exogenous testosterone drug replacement therapy is a common treatment for TDS. Studies have confirmed that patients can benefit from testosterone replacement therapy, such as increased libido, increased bone density, improved mood and cognition, and physical enhancement.
  • exogenous testosterone drug replacement therapy has obvious defects: (1) due to individual differences in testosterone levels, the dose of testosterone is difficult to master, and the dose is too small to be effective. If the dose is too large, it is prone to side effects and complications.
  • the decrease in the number of Leydig cells or hypofunction is considered to be the core pathogenesis of TDS, suggesting that cell transplantation therapy may achieve better results.
  • the development of cell transplantation therapy depends on the development of TDS model, so the research of TDS model becomes a key issue in the research field of new methods of TDS therapy.
  • EDS ethane-1,2-dimethyl sulphonate
  • LCs showed apoptosis after 24 hours, and all LCs were cleared after 72 hours.
  • Newer LCs can be found in the testicular stroma around 2 to 3 weeks, and the number of LCs is restored to the pre-dose level in about 8 to 10 weeks.
  • Serum LH gradually increased within 14 days after EDS injection, and serum LH began to decrease due to neonatal LC secretion of testosterone within 14 to 21 days. Serum LH decreased to pre-injection level after 21 days. Within 2 to 8 weeks after EDS injection, the rats lost fertility.
  • cynomolgus primate is similar to humans in terms of anatomy, physiological function and biochemical metabolism. It is an ideal experimental animal for understanding human physiology and pathophysiology, and is the most important model in clinical transformation research.
  • data from non-human primate models is essential, so the establishment of a primate TDS model is essential.
  • the non-human primate model is very expensive. How to ensure the efficient and safe establishment of animal models is the basis for the clinical transformation of human SLCs in the treatment of TDS.
  • the purpose of the present invention is to provide a safe and stable animal model of testosterone deficiency in view of the deficiencies of existing non-human primate TDS models, and provide a good basis for studying the effectiveness and safety of human SLCs in treating TDS.
  • the present invention provides a method for establishing a testosterone deficiency model of monkeys by injecting dimethyl sulfonate (EDS) into the testicular arteries of an adult monkey to block the flow of spermatic cord blood.
  • EDS dimethyl sulfonate
  • the amount of dimethyl sulfonate injected is at least 50 mg per testicle.
  • the amount of dimethyl sulfonate injected is from 50 to 200 mg per testicle.
  • the amount of dimethyl sulfonate injected is from 50 to 100 mg per testicle.
  • the amount of dimethyl sulfonate injected is from 100 to 200 mg per testicle.
  • the specific steps are as follows: after anesthetizing the adult cynomolgus monkey, disinfecting the lower abdomen and the perineum, exposing the bilateral spermatic cord with the inguinal incision, and blocking the spermatic cord blood flow with the non-invasive forceps 20-30 Minutes; the scrotal incision exposes the testes and dimethyl sulfonate is injected into the testicular artery.
  • EDS injection 0, 1, 2, 3, 4, 5, 7, 9, 11, 13, 15, 20, 25, 30, 35, 40, 45 days after EDS injection, from 9:00 am to 10:00 am, blood was collected from the upper extremity vein, each 1-2 ml of blood was allowed to stand at room temperature for 90 minutes, centrifuged at 2000 g for 15 minutes, and serum was separated and stored at -20 ° C for testosterone and liver function.
  • Testicular tissue was obtained by testicular puncture on 4, 14, 28, and 45 days after EDS injection. After fixation with 4% paraformaldehyde, the sample was dehydrated in 10%, 20%, and 30% sucrose solution to the bottom of the tissue. Buried, frozen slicer 6 ⁇ m thick sections, 5 samples per time point. Immunofluorescence staining stained the Leydig cell-specific protein CYP11A1.
  • the time to inject dimethyl sulfonate into the testicular artery is 1-5 minutes.
  • the adult monkey is at least 6 years old.
  • the method of the invention adopts an adult monkey (preferably cynomolgus monkey), and adopts a method of injecting EDS into the testicular artery to block the blood flow of the spermatic cord, and establishes a safe and stable monkey TDS model.
  • an adult monkey preferably cynomolgus monkey
  • the method can effectively remove LCs from the testes of adult male monkeys and establish a monkey TDS model.
  • the method of the present invention reduces the side effects such as liver damage caused by EDS, thereby causing the onset of TDS.
  • Mechanism studies, pharmacodynamic studies, and other preclinical studies in translational medicine provide stable model vectors.
  • Figure 1 shows the effect of intra-articular injection of EDS in combination with blocking spermatic blood flow.
  • Figure 2 is a comparison of the effects of local injection of EDS in the testes of cynomolgus monkeys and intra-articular injection of EDS to block spermatic cord blood flow.
  • Figure 3 is a comparison of the effects of injection of EDS into the testicular arteries of the cynomolgus monkey and intra-articular injection of EDS to block the spermatic blood flow.
  • Figure 4 is a testicular staining diagram.
  • Example 1 Intra-arterial injection of EDS in the testis blocked the spermatic cord blood flow, cleared the Leydig cells, and established a testosterone deficiency model.
  • the animals used in this experiment were adult male cynomolgus monkeys, age > 6 years old, weighing 8-10 kg, a total of 8 adult male cynomolgus monkeys, randomly divided into 4 groups, 2 in each group.
  • the first group (also referred to as "solvent group") 2 cynomolgus monkeys were injected intraarticularly with vehicle (DMSO: H2O, 1:3, V/V). EDS was dissolved in vehicle, the second group of 2 cynomolgus monkeys were intra-articularly injected with 50 mg EDS; the third group of 2 cynomolgus monkeys were intra-articularly injected with 100 mg of EDS; the fourth group of 2 cynomolgus monkeys were injected with 200 mg of testicular artery. EDS.
  • the cynomolgus monkey was anesthetized, the lower abdomen and the perineum were disinfected, and the bilateral spermatic cords were exposed by the inguinal incision.
  • the spermatic cord blood flow was blocked with a non-invasive forceps for 20-30 minutes; the scrotal incision exposed the testes, and the corresponding dose of EDS Slowly inject into the testicular artery for an injection time of 1-5 minutes.
  • EDS injection 0, 1, 2, 3, 4, 5, 7, 9, 11, 13, 15, 20, 25, 30, 35, 40, 45 days after EDS injection, from 9:00 am to 10:00 am, blood was collected from the upper extremity vein, each 1-2 ml of blood was allowed to stand at room temperature for 90 minutes, centrifuged at 2000 g for 15 minutes, and serum was separated and stored at -20 ° C for testosterone and liver function.
  • Testicular intra-arterial injection of 50, 100, 200mg/testis EDS combined with blocking spermatic cord blood flow can significantly reduce serum testosterone levels in cynomolgus monkeys.
  • Testosterone levels in the 50 mg/testis group were reduced to 30% of normal levels, and testosterone levels in the 100 and 200 mg/testis groups were below the lower limit of detection of 0.13 ng/ml.
  • Testosterone levels in the 50, 100, 200 mg/testis group returned to pre-dose levels 45 days after EDS injection.
  • ALT and AST of the liver group showed no significant changes in liver damage, and were at normal levels; 1 day after EDS injection, ALT and AST increased in 50, 100, 200 mg/testis group, and the dose The increase was increasing and returned to pre-dose levels within 7 days.
  • Testicular tissue was obtained by testicular puncture on 4, 14, 28, and 45 days after EDS injection. After fixation with 4% paraformaldehyde, the sample was dehydrated in 10%, 20%, and 30% sucrose solution to the bottom of the tissue. Buried, frozen slicer 6 ⁇ m thick sections, 5 samples per time point. Immunofluorescence staining stained the Leydig cell-specific protein CYP11A1.
  • Figure 4 is a testicular staining diagram. Among them, A: testicular staining on the 4th day after EDS injection, the test shows that the Leydig cells were completely cleared; B: Testicular staining on the 14th day after EDS injection, the test shows that the Leydig cells began to recover; C: EDS Testicular staining on the 28th day after injection, the figure shows that the number of Leydig cells was restored by about 50%; D: Testicular staining on the 45th day after EDS injection, the figure showed that the number of Leydig cells was restored to normal. Testicular sections show that EDS only affects Leydig cells, but does not affect other cells.
  • Example 2 Comparison of local injection of EDS in the testes of cynomolgus monkeys and intra-articular injection of EDS to block spermatic blood flow.
  • the cynomolgus monkey On the day of the experiment, the cynomolgus monkey was anesthetized and disinfected in the lower abdomen and perineum.
  • the first group also known as the "local group” was injected with 100 mg/testis EDS in two testes of the two cynomolgus monkeys; the second group (also called " The two groups of cynomolgus monkeys exposed the bilateral spermatic cords with an inguinal incision, and the spermatic cord blood flow was blocked with a non-invasive forceps for 20-30 minutes; the scrotal incision exposed the testicles, and an equal amount of EDS was slowly injected into the testicular artery. The injection time is 1-5 minutes.
  • EDS injection 0, 1, 2, 3, 4, 5, 7, 9, 11, 13, 15, 20, 25, 30, 35, 40, 45 days after EDS injection, from 9:00 am to 10:00 am, blood was collected from the upper extremity vein, each 1-2 ml of blood was allowed to stand at room temperature for 90 minutes, centrifuged at 2000 g for 15 minutes, and serum was separated and stored at -20 ° C for testosterone and liver function.
  • EDS testicular local injection of EDS group (local group) testosterone level was not affected
  • testicular intra-arterial injection of EDS + block spermatic cord blood flow group (combination group) cynomolgus monkey serum testosterone level is lower than detection
  • the lower limit was 0.13 ng/ml and returned to pre-dose levels 45 days after EDS.
  • B, C After EDS injection, the local group showed no significant changes in liver damage index ALT, AST, at normal level; 1 day after EDS injection, the combined group ALT, AST mildly increased, and returned to administration within 4 days Pre-level.
  • Example 3 Comparison of the effect of injection of EDS and testicular intra-articular injection of EDS on the spermatic cord blood flow in cynomolgus monkey testicular arteries.
  • the cynomolgus monkey On the day of the experiment, the cynomolgus monkey was anesthetized and disinfected in the lower abdomen and perineum.
  • the first group of 2 cynomolgus monkeys were injected with 100 mg/testis EDS in both testicular arteries; the second group (also called “combined group”) was treated with the inguinal inner ring.
  • the incision exposes the bilateral spermatic cord, and the spermatic cord blood flow is blocked with a non-invasive forceps for 20-30 minutes; the testicular is exposed by the scrotal incision, and an equal amount of EDS is slowly injected into the testicular artery for an injection time of 1-5 minutes.
  • EDS injection 0, 1, 2, 3, 4, 5, 7, 9, 11, 13, 15, 20, 25, 30, 35, 40, 45 days after EDS injection, from 9:00 am to 10:00 am, blood was collected from the upper extremity vein, each 1-2 ml of blood was allowed to stand at room temperature for 90 minutes, centrifuged at 2000 g for 15 minutes, and serum was separated and stored at -20 ° C for testosterone and liver function.
  • the method of the present invention blocks the spermatic blood flow by EDS injection in the testicular arteries of cynomolgus monkeys, and continuously and long-term observation of liver and other indicators, and proves that the method can effectively remove LCs from the testes of adult male cynomolgus monkeys.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Environmental Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Emergency Medicine (AREA)
  • Zoology (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Animal Husbandry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé d'établissement d'un modèle simiesque de déficience en testostérone, celui-ci adoptant le procédé d'injection d'acide diméthyl-éthanesulfonique dans l'artère testiculaire de singes adultes en combinaison avec le blocage du flux sanguin dans le cordon spermatique. Le procédé fournit un modèle animal de déficience en testostérone sûr et stable.
PCT/CN2018/123563 2018-02-24 2018-12-25 Procédé d'établissement d'un modèle simiesque de déficience en testostérone Ceased WO2019161707A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810157030.1 2018-02-24
CN201810157030.1A CN108524485A (zh) 2018-02-24 2018-02-24 一种猴睾酮缺乏模型的建立方法

Publications (1)

Publication Number Publication Date
WO2019161707A1 true WO2019161707A1 (fr) 2019-08-29

Family

ID=63485780

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/123563 Ceased WO2019161707A1 (fr) 2018-02-24 2018-12-25 Procédé d'établissement d'un modèle simiesque de déficience en testostérone

Country Status (2)

Country Link
CN (1) CN108524485A (fr)
WO (1) WO2019161707A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108524485A (zh) * 2018-02-24 2018-09-14 中山大学 一种猴睾酮缺乏模型的建立方法
CN110257433B (zh) * 2019-05-14 2022-08-16 中山大学 酪氨酸血症i型猴模型及其建立方法和应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102883710A (zh) * 2010-04-12 2013-01-16 克劳拉斯医疗有限公司 口服睾酮酯制剂以及包含其的治疗睾酮缺乏症的方法
CN103118684A (zh) * 2010-08-16 2013-05-22 英诺泰斯托比利时公司 用于治疗睾酮缺乏的睾酮溶液
CN105255826A (zh) * 2015-11-27 2016-01-20 中山大学 人诱导多功能干细胞向睾丸间质细胞的诱导分化方法及其用途
CN108524485A (zh) * 2018-02-24 2018-09-14 中山大学 一种猴睾酮缺乏模型的建立方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102883710A (zh) * 2010-04-12 2013-01-16 克劳拉斯医疗有限公司 口服睾酮酯制剂以及包含其的治疗睾酮缺乏症的方法
CN103118684A (zh) * 2010-08-16 2013-05-22 英诺泰斯托比利时公司 用于治疗睾酮缺乏的睾酮溶液
CN105255826A (zh) * 2015-11-27 2016-01-20 中山大学 人诱导多功能干细胞向睾丸间质细胞的诱导分化方法及其用途
CN108524485A (zh) * 2018-02-24 2018-09-14 中山大学 一种猴睾酮缺乏模型的建立方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MA, XUE ET AL.: "Effects of Ethane Dimethane Sulfonate on Fetal Leydig Cells in Neonatal Rats", JOURNAL OF PEKING UNIVERSITY (HEALTH SCIENCES), vol. 45, no. 5, 31 October 2013 (2013-10-31), pages 770 - 773, XP055633422, ISSN: 1671-167X *
SUMATHI, R.: "Ethane dimethylsulphonate selectively destroys Leydig cells in the adult bonnet monkeys (Macaca radiata", ASIAN JOURNAL OF ANDROLOGY, vol. 1, no. 3, 31 January 1999 (1999-01-31), pages 115 - 120, XP055633415, ISSN: 1008-682X, Retrieved from the Internet <URL:http://www.asiaandro.com/archive/1008-682X/1/115.htm> *

Also Published As

Publication number Publication date
CN108524485A (zh) 2018-09-14

Similar Documents

Publication Publication Date Title
Plemel et al. Remyelination after spinal cord injury: is it a target for repair?
Yang et al. JAK 2/STAT 3 activation by melatonin attenuates the mitochondrial oxidative damage induced by myocardial ischemia/reperfusion injury
CN102497873B (zh) 用于增加生存年限和健康年限的组合物和方法
Montagnese et al. Structural plasticity in the rat supraoptic nucleus during gestation, post-partum lactation and suckling-induced pseudogestation and lactation
Chen et al. Treatment with enriched environment reduces neuronal apoptosis in the periinfarct cortex after cerebral ischemia/reperfusion injury
Shen et al. Cortical spreading depression preconditioning mediates neuroprotection against ischemic stroke by inducing AMP‐activated protein kinase‐dependent autophagy in a rat cerebral ischemic/reperfusion injury model
THORN et al. Pheochromocytoma of the adrenal associated with persistent hypertension; case report
Xu et al. Protective effect of lithium chloride against hypoglycemia-induced apoptosis in neuronal PC12 cell
CN112494479B (zh) 茶黄素在制备卵巢功能保护药物方面的应用
WO2019161707A1 (fr) Procédé d&#39;établissement d&#39;un modèle simiesque de déficience en testostérone
Tang et al. Hypoxic preconditioned mesenchymal stem cells ameliorate rat brain injury after cardiopulmonary resuscitation by suppressing neuronal pyroptosis
Li et al. Reproductive and developmental toxicity assessment of human umbilical cord mesenchymal stem cells in rats
Liu et al. The effects of human umbilical cord mesenchymal stem cell transplantation on female fertility restoration in mice
CN112716953B (zh) Cdn1163在制备缓解或者治疗神经性疼痛的药物中的应用
KR20180035739A (ko) 조직 재생 및 저하된 조직 기능의 회복을 자극하기 위한 제제로서의 디카르복시산의 비스아미드 유도체
CN115137715A (zh) 一种姜黄素在制备治疗早发性卵巢功能不全及卵巢响应匮乏药物中的应用
Sun et al. The potential role of melatonin on mental disorders: insights from physiology and pharmacology
CN101584686B (zh) 丙戊酸钠在制备治疗视网膜疾病的药物中的应用
WO2025152870A1 (fr) Utilisation de finérénone dans la préparation d&#39;un médicament pour le traitement et/ou la prévention de maladies de la dysfonction ovarienne
CN114984028B (zh) 黄芪甲苷的用途
Zhang et al. Effect of combination therapy with neural stem cell transplantation and teramethylpyrazine in rats following acute spinal cord injury
Yeni et al. Effect of Curcumin in Decreasing MDA Level in Preeclampsia-Induced Human Umbilical Vein Endothelial Cell (HUVEC)
KR20140041263A (ko) 말초혈액 단핵세포를 유효성분으로 포함하는 예방 또는 치료용 약제학적 조성물
CN114010660A (zh) 一种ASP联合hAD-MSCs治疗POI的实验方法
Aljuboury et al. Effect of Hydrocortisone on Spermatogenesis in Male Rats

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18906870

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18906870

Country of ref document: EP

Kind code of ref document: A1