Nghiên cứu tính an toàn và tác dụng cải thiện khả năng sinh tinh của viên nang y10 tt tiếng anh

INTRODUCTION Male infertility is a disease that greatly affects the society WHO predicted it would be the rd common dangerous disease in 21 st century after cancer and heart diseases Infertility couples in the world accounts for 15% and the figure for Vietnam is 8% Causes of male infertility are enormous, but the most frequent is sperm decline Modern medicine has gained achievements in infertility due to sperm decline treatment; however, it also presents side effects Traditional oriental medicine that has experienced thousands of years of experience has also had specific male infertility treatments and brought good results Many scientific studies have proven that traditional medicine treatments have good effects on the treatment of male infertility in general and sperm decline in particular Cornu cervi parvum and Cordyceps militaris cultured in Vietnam are two valuable medicinal herbs, which are widely used in the treatment of infertility and male reproductive dysfunction, with good potential However, there have been are no scientific studies on the effects of Cornu cervi parvum and cordyceps and their therapeutic effects on treating reproductive functions We carry out the research with the following objectives: Assess the safety and effects on sperm production of Y10 on experimental animals Assess the effects on the sperm production of Y10 on patients with sperm decline PRACTICAL MEANINGS AND NEW CONTRIBUTIONS OF THE DISSERTATION The reproductive health in Vietnam has changed significantly in recent years The rate of infertile couples is increasing, especially infertility due to sperm decline in men Modern medical treatments have shown a positive role and effectiveness in the treatment of sperm decline, but it can cause physiological trauma and side effects Thus, it is always necessary to continue the search for effective, economical and safe herbal-based medicines, it is also the research direction that scientists in the country and in the world are interested in The dissertation is systematic and experimental, and scientific work The research and its application on the treatment of male infertility due to sperm decline by traditional remedies has contributed to elucidate the theory of traditional medicine and gradually modernize it, which has scientific and practical values STRUCTURE OF THE DISSERTATION The dissertation consists of 126 pages: 02 pages of introduction, 39 pages of literature review, 22 pages of subjects and research methods, 34 pages of research results, 25 pages of discussion, 03 pages of conclusion and 01 page of recommendation The thesis has 141 references (42 in Vietnamese, 64 in English, and 35 in Chinese), 45 tables, charts, 01 diagram, and 15 photos and appendices Chapter LITERATURE REVIEW 1.1 The view of modern medicine on sperm decline 1.1.1 The causes of sperm decline Sperm decline is caused by factors that affect sperm growth and disrupt that process There are main types of causes, as followed: * Reproductive hormonal disorders: Some reproductive hormones play a decisive role in sperm production in general and its single differentiation stage in particular Diseases caused by reproductive hormonal disorders lead to a huge change in the sperm count and quality * Genetic causes: testicular, microbiological and temperature factors; varicose veins of the spermatic cord damage, X-rays, radiation and chemicals, PH of semen, or medical drugs might cause sperm decline, etc * Other causes: diet, immune, living and working environment, or stress 1.1.2 Directions of treatment of sperm decline according to modern medicine * Using modern medicine: Anti-oxidants: Glutathion, L-arginin, vitamin C; Hormones: FSH (humegon, puregon), LH (pregnyl), hMG or hCG; andriol testocaps, mesterolone, testosterone undecanoate; Corticoids are used for infertility due to anti-sperm antibodies; Antibiotics are indicated for urinary and genital infections The common bacteria are Chlamydia trachomatis, E.coli, and U Urealyticum - Using of traditional medicine: the trend of research using alternative therapies and traditional medicine to treat sperm decline is increasing - Surgery: surgery should be indicated for patients with impaired spermatozoa that affect the sex life, such as varicose veins, ectopic testicles, testicular water or inguinal hernia Connective surgery should be indicated for patients with no sperms by connecting vas deferens IUI: a method of putting a small volume of sperm that has been selected by sperm washing technique into the uterus near the time of ovulation; IVF and embryo transfer into the uterus: a technique for in vitro fertilization ICSI: sperm injection directly into the ovule to create embryos 1.2 Traditional medicine theory on sperm decline 1.2.1 Traditional medicine theory on male genital and reproductive functions Traditional medicine does not have the name of sperm impairment but the disease is classified as impotence in modern medicine 1.2.2 Causes and mechanism of pathogenesis of sperm decline according to traditional medicine Traditional medicine suggests that the causes of emotions, climatical conditions in excess, trauma, or long-term internal injuries will affect the viscera organs, causing physiological disorders of the viscera organs, thereby leading to failure sperm decline and infertility 1.2.3 Treatment of sperm decline according to Traditional Medicine Traditional methods of treatment of sperm decline include traditional medicine, acupuncture, elec-accupuncture, macrobiotic medicine, or a combination of the methods for the most effective treatment * Medication therapies Deficiency of Kidney and Jing: Therapy: warming Kidney and strenthening yang Formulas: wu zi yan zong wan in combination with gui wan Ingredients:Aconitum Camichaeli Debx.,Laurus cassia L, Codonopsis pilosula , Astragalus propinquus, Rehmannia glutinosa Libosch, Dioscorea opposita, Cornaceae , Cortex Moutan,-Cortex Paeoniae Suffuticosae, Alisma plantago aquatic L., Poria cocos Wolf.,Zingiber officinale, Glycyrrhiza uralensis, and Glycyrrhiza uralensis Deficiency of Gate of life Fire: Therapies: warming kidneys, strengtening Yang Nourishing kidneys to promote Jing production Formulas: jin gui shen qi wan in combination with bao yuan tang Ingredients: Aconitum fortunei Hemsl., Cinnamomum cassia Nees & Eberth, Codonopsis pilosula (Franch) Nannf., Astragalus propinquus., Rehmannia glutinosa (Gaertn.) Libosch., Dioscorea persimilis Prain et Burk., Cornus controversa., Paeonia suffruticosa Andr., Alisma plantago aquatica L, Poria cocos Wolf., Zingiber officinale, and Glycyrrhiza uralensis Deficiency of spleens and Jing: Therapies:Warming and tonifying spleens and kidneys Toniffying Qi and promoting the production of Jing Formulas: shi zi tang in combination with iu jun zi tang Ingredients: Cuscutasinesis – Lamk, Morus Alba L , Lycium barbarumL., Ligustrum lucidum Ait., Selinum monnieri L., Rubus idaeus, Rosa laevigata Michx., Fructus Schisandra Deficiency of Qi and blood: Therapies: Tonifying Qi and nouishing blood Tonifying kidney to stimulate the Jing production Formulas: yi jing zi yu tang in combination with ba junzi tang Ingredients: Placenta Hominis, Cornaceae, Codonopsis pilosula, Astragalus propinquus, Atractylodes macrocephala, Wolfiporia extensa, Dioscorea persimilis Prain et Burk, Glycyrrhiza uralensis, Angelica sinensis, Paeonia lactiflora Pall., Rehmannia glutinosa Libosch, Lycium barbarumL., Gynochthodes officinalis, and Epimedium Stagnation of liver qi: Therapies: Dispersing the stagned liver qi Activating Qi and blood circulation to promote the Jing production Formulas: Ingredients: xue fu zhu yu tang Prunus persica Stokes, Carthamus tinctorius, Paeonia liacliflora Pall, Ligusticum striatum, Angelica sinensis, Bupleurum chinesnis DC., Liquidambar formosana Hance, and Squama Manidis Wetness heat evil attacking the lower warmer: Therapies: Clearing heat Eliminating dampness Expelling toxic Formulas: ong dan zuo gan tang in combination with liu wei di huang wan Ingredients: Gentiana scabra Bunge., Phellodendron amurense, Gardenia jasminoides ellis, Lonicera japonica Thunb., Forsythia suspensa, Semen Plantaginis, Alisma plantago-aquatica, Rehmannia glutinosa Libosch, Cornaceae, Dioscorea persimilis Prain et Burk., and Cortex MoutanCortex Paeoniae Suffuticosae * Alternative therapies Common therapies are accupuncture and elec-accupuncture which are often used together to get highest effectiveness * Accupunture: Use the ponts of Ming men, Yao yuan guan (Du.3) , Guan yuan (Ren.4), Zhong ji (Ren.3), San jin jiao (Sp 6), Zhi shi (UB.52), Thai yuan (Lu.9), Zu san li (St.36), etc 1.5 Overview of Cornu cervi parvum and Cordyceps militaris Cornu cervi parvum: Characteristics: Sweet and salt taste Warm Benefit liver and kidney meridians Effects: Strengthen Yang Nourishing Jing blood Strengthening bones Regulating meridian points It is used in cases of kidney Yang imbalance, deficiency of Jing blood, impotence, fatigue, dizziness, back ache, etc Cordyceps militaris: Characteristics: Sweet Neutral Benefit lung and kidney meridians Effects: According to ‘ben cao gang mu shi yi’ the herb maintains lung Qi, nourishes kidneys and good for oldage In traditional medicine, Cornu cervi parvum Cordyceps militaris are two precious herbs that tonify kidneys and nourish Jing When they are combined, they would be beneficial for both kidney Yang and Yin, Jing and Qi They, hence, have been used for longs in male infertility effectively They are soaked in wine Chapter SUBJECTS AND METHODS OF THE STUDY 2.1 Research material Studied madication: Y10 made from Cornu cervi parvum purchased in Huong Son - Ha Tinh and Cordyceps militaris cultured at Military Medical Academy, meeting its standards 2.2 Research subjects Studied animals: Adult white mice, purebred Swiss lines, sexes, weight 18 - 22g, total of 470 mice Adult white rats, purebred Wistar lines, both sexes, weighing 160 - 180g, a total of 80 rats Studied patients: 30 military men with sperm examined at the Embryo Technology Training and Research Center - Military Medical Academy They were consistent with disease selection and exclusion criteria according to traditional medicine and modern medicine, and volunteered to the research 2.3 Location and time The experimental research was conducted at the Department of Clinical Pharmacy - Military Medical Academy, from May 2017 to October 2017 The clinical research was conducted at the Embryo Technology Training and Research Center - Military Medical Academy, from October 2017 to December 2017 2.4 Research Methods 2.4.1 The study on the safety and effects of Y10 on the experimental animals * The acute and semi-chronic toxicity study The acute toxicity was determined in accordance with the guidelines of the Ministry of Health and the OECD by oral route with increasing doses LD50 was calculated by the Litchfield-Wilcoxon method The semi-chronic toxicity was assessed according to the regulations of the Ministry of Health of Vietnam and the guidance of OECD, WHO 30 mice were divided into groups, 10 each; the control group drank distilled water Study group 1: Y10 224mg decoction / kg / day Study group 2: Y10 672mg decoction / kg / day in 90 consecutive days Their general condition, body weight, hematopoietic function, liver and kidney function, liver and kidney histopathology were observed and recorded before and after the treatment * The study of toxicity on reproductive abiliity of Y10 The white mice of both sexes were randomly divided into study groups, with 60 each including 20 males and 40 females The control group consisted of 20 males and 40 females drinking distilled water -Group 1: 20 males and 40 females taking Y10 384mg decoction/kg/ day - Group 2: 20 males and 40 females taking Y10 1152mg decoction / kg / day -Group 3: 20 males taking Y10 384mg decoction / kg / day and 40 females taking distilled water - Group 4: 20 males taking Y10 1152mg decoction / kg / day and 40 females taking distilled water The treatment duration was 60 days, at a certain time everyday (8 am) After 60 days, a male was kept with two females in a separate cage (generation P) and the reproduction process was monitored - The conception and fetal development were monitored * The mutagenic toxicity study on chromosomes of Y10 on white mice According to OECD guideline 475 (2002), 90 healthy white mice, divided into groups, each group of 30 rats including 15 male rats and 15 female rats (male and female rats kept separately): + Group (control group): distilled water + Group 2: Y10 384mg dectoction / kg / day + Group 3: Y10 1152mg decoction / kg / day Mice were given the medication daily, once a day at am, in weeks (28 days) After the groups finished drinking the decoction, they were injected with colcemide into the abdominal cavity to stop cell division in the middle of cell division, when the chromosomes were most typical hours after the colcemide injection, the chromosome samples from the bone marrow were made by Ford method and the chromosomes from the testes by Evan method 50 chromosome patterns of each mouse were analyzed at the Diakinesis-metaphase stage when the chromosomes were short, evenly sprung and were not overlapped * Evaluation of the effects of sperm enhancement of Y10 on white rats that caused sperm decline with Natri valproate Adult male rats, randomly assigned to five study groups, 10 each + Group 1: were not caused sperm decline, drank distilled water + Group 2: caused sperm decline, drank distilled water + Group 3: caused sperm decline, drank testosteron undecanoat 16 mg / kg / day + Group (batch 1): caused sperm decline, drank Y10 224 mg decoction / kg / day + Group (batch 2): caused sperm decline, drank Y10 448mg decoction / kg / day The group of mice that were caused sperm decline by Natri valproate at the dose of 500mg / kg / day in weeks Mice in non-sperm-declinecaused groups were given distilled water at the same volume in weeks After weeks, mice were killed to serve as samples for the evaluation of research indicators, including: testosteron, sperm density, sperm motility, sperm morphology, percentage of structural morphology sperm abnormalities, testicular histopathological changes, size of spermatogenesis diameter, genital organs (testes, epididymis, seminal vesicles, Cowper glands, glans, prostate gland, and levator ani muscles) The genital organs were determined out of every 100g the mice’s body weight 2.4.2 Clinical research We conducted inclusive studies and open clinical trials, compared the differences in the indexes between before and after the treatment The patients with sperm decline were examined according to modern and traditional medicine, they undertook all tests and if they were eligible, they would be selected for the study groups * Criteria for selecting patients 30 patients (male soldiers) were selected based on the following criteria: Patients who voluntarily took part in the research, stopped using drugs that affect sperm count and quality for at least 75 days - Criteria for selecting patients according to modern medicine: Age: 16 to 56; Sperm decline according to WHO standards onseminogram (2010) - Criteria for selecting patients according to traditional medicine: men had ‘deficiency of kidney and Jing’ - Criteria for excluding patients: The patients who did not agree to take part in the study, did not strictly followed guidances, or refused to continue the treatment * Dosage: tablets per day, twice, hours after eating, in months * Methods of clinical examination and clinical monitoring The medical record for each patient is made according to a unified form based on the criteria of the World Association of Southern Studies, combined with với four methods for physical examination of traditional medicine, the patients were examined and their test results were recorded * Laboratory tests - Biochemical blood before and after the treatment: urea, creatinine, AST, ALT - Quantification of LH, FSH, and serum testosterones before the treatment; quantification of LH, FSH, and serum testosteron in selected patients after the treatment - Seminogram before and after the treatment The tests were done at the Institute of Medical Research - Military Medical Academy * Evaluation criteria - Epidemiological characteristics: distribution of patients by age and types of infertility (infertility I or infertility II) - Some clinical signs due to unwanted effects of the medication: rashes, digestive disorders (loose stool, constipation , etc), dizziness, etc - Clinical symptoms due to renal impairment before and after the treatment - ALT, AST, urea, serum creatinine before and after the treatment - Serum testosteron, LH, FSH concentrations before and after the treatment - Semen before and after treatment (Table 2.3) - The percentage of patients whose wives were pregnant and gave birth after the treatment * Data processing The research data is processed by biomedical statistical method, using statistical software SPSS.17.0 The difference was statistically significant when p 0.05) * Changes in histopathology: macroscope morphology images of liver organs, spleens, and kidneys of rats in research groups and had even dark reddish brown color, smooth surface, no lumps or hemorrhages When being pressed down, there were no differences compared to that of the control group 3.1.3 Results of the toxicity study on fertility Table 3.1 The percentage of female mice conceived in the groups Groups Control group Group Group Group Group P generation % female mice p conceived 60.94 % 59.86 % 63.28 % 61.72 % 66.37% F1 generation % female mice conceived p 71.65 % > 0.05 70.14 % 74.56 % 72.43 % 77.91% > 0.05 Comments: There was no difference in the pregnancy rate between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.2: The number of corpus luteum / mother mouse in the groups Groups Control group Group Group Group Group P generation corpus luteum / p mother mouse corpus luteum / mother mouse p 13.64 ± 2.57 12.36 ±2.08 12.45±3.14 12.18±2.56 13.09±2.81 12.27±2.35 F1 generation > 0.05 14.22 ±2.86 13.69±3,02 14.06 ±2.65 13.91 ±3.14 > 0.05 Comments: There was no difference in the average number of corpus luteum / mother rat between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.3: Number of alive fetus / mother mouse in the groups(%) Groups Control group Group Group Group Group P generation Number of alive fetus / mother mouse F1 generation Number of alive fetus / mother mouse p 97.65 % 96.92% 98.45% 97.26 % 98.19% p 98.02% > 0.05 97.65% 98.69% 97.91% 98.54% > 0.05 Comments: There was no difference in the number of pregnant fetuses / mother rat between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.4: Number of early fetal deaths / female mouse in the groups(%) Groups Control group Group Group Group Group P generation Number of early fetal deaths / female mouse p 2.94 % 3.16 % 2.08 % 2.75 % 3.21 % F1 generation Number of early fetal deaths / female mouse p 3.62 % > 0.05 3.09% 4.17% 2.98% 3.81% > 0.05 Comments: There is no difference in the number of early fetal death / mother rat between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.5: Number of stillbirths / mother mouse in each group (%) Groups Control group Group P generation Number of stillbirths / p mother mouse 1.62 % F1 generation Number of stillbirths / mother mouse p 2.36% 1.81% 2.09% > 0.05 Group 2.03% 1.86% Group 2.16% 2.47% Group 1.25% 2.18% > 0.05 Comments: There was no difference in the number of stillbirth fetal fetuses / female rat between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.6: Number of dissipated eggs / mother mouse in the groups (%) P generation Groups Number of dissipated eggs / mother mouse Control group Group p F1 generation Number of dissipated eggs / mother mouse 4.48% 4.62% p 3.96% > 0.05 > 0.05 4.12% Group 4.26% 3.97% Group 4.32% 4.31% Group 3.98% 2.68% Comments: There was no difference in the number of dissipated eggs / mother between the Y10 and control groups in P and F1 generations (p> 0.05) Table 3.7: Number of mice per a litter Groups Control group Group Group Group Group F1 generation Number of mice per a litter p 12.08 ±1.93 12.14 ± 2.54 12.19± 1.98 12.23 ± 1.64 12.45± 2.46 > 0.05 Comments: There was no difference in the number of mice / litter between the Y10 and control groups in P and F1 generations (p> 0.05) Because the proportion of male and female mice in each litter (F1 generation) in each group was equal, the number of mice per litter in groups was similar We randomly selected the number of female mice and the number of male mice in the groups so that the number of pairs in the group was the same Table 3.8: Number of dead mice per litter (%) Groups F1 generation Number of dead mice per litter p Control group Group Group Group Group 1.68 % 2.08 % 1.98% 1.62 % 2.15% > 0.05 Comments: Mice born to F1 generation mother mice: acted and moved normally No deformities in all groups Conclusion: Results of the genetic toxicity study (reproductive toxicity) showed that Y10 capsules did not significantly affect the normal development of fetus and mices born to P and F1 generations 3.1.4 Research results on chromosomal toxicity Table 3.9 The effects of Y10 on the number of bone marrow cell chromosomes Indexes Number of cells monitored Number of Aneuploid Rate of Aneuploid (%) Number of polyploidy Rate of polyploidy (%) Group 172 2.05 3.07 Groups Group 169 1.18 2.96 Group 160 1.25 1.88 p > 0.05 > 0.05 Comments: on chromosomal samples from bone marrow cells in groups taking preparations at both low and high doses continuously in 28 days, the rate of occurrence of chromosomal disorders was not different from that of the control group (p> 0.05) Table 3.10 Effect of Y10 on bone marrow cell chromosome structure Indexes Number of cells monitored Number of chromosomal chromatid disorders Rate of chromosomal chromatid disorders (%) Number of chromosomal structural disorders Rate of chromosomal structural disorders (%) Rate of chromosome cluster disorders Group 172 Group 169 Grou p3 160 0 1.12 0.00 0.00 0 1.69 0.00 0.00 0 p > 0.05 > 0.05 Comments: The rate of chromosomal structural disorders in the chromosomal samples of bone marrow cells in the mice taking the decoction at low and high doses did not differ from that of the control group (p> 0.05) Control group Low dose group Photo 3.5 White bone marrow cell chromosome s (X 1000) After the groups finished drinking the decoction, they were injected with colcemide into the abdominal cavity to stop cell division in the middle of cell division, when the chromosomes were most typical hours after the colcemide injection, the chromosome samples from the bone marrow was made by Ford method and the chromosomes from the testes by Evan method 50 chromosome patterns of each mouse were analyzed at the Diakinesis-metaphase stage when the chromosomes were short, evenly sprung and were not overlapped Table 3.11 Effects of the decoction on testicular chromosomes Types of mutations Number of chromosome < 40% Number of chromosome = 40% Number of chromosome > 40% Number of normal chromosome % Number of gender chromosome % Control group Group 6.48 ± 0.84 7.04 ± 0.80 90.87 ± 1.32 90.98 ± 1.43 1.23 ± 0.64 1.14 ± 0.54 1.25 ± 0.67 1.19 ± 0.59 7.82 ± 1.41 8.23 ± 1.52 Group 7.30 ± 1.01 89.50 ± 0.65 1.08 ± 0.61 1.10 ± 0.75 8.90 ± 1.39 p > 0.05 > 0.05 > 0.05 > 0.05 > 0.05 Comments: there was no difference in the frequency of chromosomal mutations of the testes between the study and control groups (p> 0.05) Conclusion: The Y10 did not cause chromosomal mutations in bone marrow and testes at the doses and times in the experiment 3.2 Research on the effects of increasing sperm function of Y10 in the experiment 3.2.1 Effects of Y10 on mice’s serum testosterone Table 3.12 Ratio serum testosterone Hi Group Testosteron (ng/ml) 3.51 ± 2.10 Group 1.55 ± 0.68 ↓ 55.95* % Group 3.10 ± 1.27 ↑ 99.81** % Group 2.77 ± 1.27 ↑ 78.71** % Group 2.94 ± 1.38 ↑ 89.35** % Groups % changes p p1.3.4.5-2< 0.01 p3.4.5-1> 0.05 p4.5-3> 0.05 p4-5> 0.05 Comments: The serum testosteron concentrations in the groups 3, 4, increased significantly compared to the group (p 0, 05) There was no significant difference between the groups 3, 4, (p> 0.05) 3.2.2 Effects of Y10 on sperm count and quality Table 3.13 Effects of Y10 on sperm density Groups Group Group Group Group Group sperm density (× 106/mL) 81.58 ± 23.99 39.14 ± 11.90 69.11 ± 23.31 79.09 ± 15.44 82.45 ± 14.69 % changes ↓ 52.03* % ↑ 76.57**% ↑ 102.09**% ↑ 110.67**% p p1.3.4.5-2< 0.01 p3.4.5-1> 0.05 p4.5-3> 0.05 p4-5> 0.05 Comments: The sperm density of the group decreased significantly compared to that of the group 1, the difference was statistically significant with p 0.05 Rate of Sperm mobility (%) Slow Nonprogressive progressive 4.47 ± 1.54 6.34 ± 1.60 9.13 ± 4.10 9.43 ± 3.39 5.46 ± 1.48 6.38 ± 1.60 4.60 ± 1.61 6.94 ± 2.17 4.95 ± 1.54 6.47 ± 1.50 p-2< 0.05 p-2< 0.05 p3.4.5-1> 0.05 p3.4.5-1> 0.05 Non-mobile 51.00 ± 9.02 60.32 ± 9.42 53.59 ± 9.15 53.72 ± 9.72 53.08 ±10.19 p-2< 0.05 p3.4.5-1> 0.05 p4.5-3> 0.05 p4-5> 0.05 p4.5-3> 0.05 p4-5> 0.05 p4.5-3> 0.05 p4-5> 0.05 p4.5-3> 0.05 p4-5> 0.05 Comments: Mice in the groups 3, and had significantly higher rates of rapid progressive sperms than the group (p 0.05); meanwhile, the percentages of sperms that did not progress and progress slowly decreased significantly compared to the group (p 0.05 p4.5-3> 0.05 p4-5> 0.05 * compared to the control group ** compared to the group Comments: The percentage of sperms having abnormal structural morphology in the groups 3, 4, decreased significantly compared to that of the group (p 0.05) The figures for the groups 3, 4, were higher than that of the groups using Y10, but there was no statistically significant difference (p> 0.05) ) 3.2.3 Effects of Y10 on the weight of male genital organs Table 3.16 Weight of the genital organs (n = 10) Weight of the genital organs (g/100g thể trọng) Lô nghiên cứu Group Group semin Prosta Cowp Epidid al Testicle te er ymis vesicle gland gland s 0.221 0.032 X 0.889 ± 0.252 ± 0.119 ± ± ± ± 0.165 0.031 0.030 0.062 0.021 SD 0.159 0.098 0.026 X 0.682 ± 0.213 ± ± ± ± ± 0.174 0.024 0.028 0.015 0.018 SD p2-1 < 0.05 < 0.05 < 0.05 < 0.05 > 0.05 Levat or ani Glans muscl es 0.036 0.326 ± ± 0.019 0.069 0.035 0.291 ± ± 0.016 0.038 > 0.05 < 0.05 X Group ± SD p3-1 p3-2 X Group ± SD p4-1 p4-2 X Group ± SD p5-1 p5-2 0.831± 0.247 ± 0.206 0.032 > 0.05 < 0.05 > 0.05 < 0.05 0.832 ± 0.250 ± 0.201 0.103 > 0.05 < 0.05 > 0.05 < 0.05 0.834 ± 0.252 ± 0.194 0.081 > 0.05 < 0.05 > 0.05 < 0.05 0.208 ± 0.030 > 0.05 < 0.05 0.210 ± 0.041 > 0.05 < 0.05 0.211 ± 0.062 > 0.05 < 0.05 0.113 ± 0.028 > 0.05 < 0.05 0.114 ± 0.038 > 0.05 < 0.05 0.115 ± 0.046 > 0.05 < 0.05 0.027 ± 0.016 > 0.05 > 0.05 0.028 ± 0.012 > 0.05 > 0.05 0.026 ± 0.014 > 0.05 > 0.05 0.036 ± 0.020 > 0.05 > 0.05 0.035 ± 0.020 > 0.05 > 0.05 0.036 ± 0.019 > 0.05 > 0.05 0.328 ± 0.058 > 0.05 < 0.05 0.325 ± 0.126 > 0.05 < 0.05 0.330 ± 0.107 > 0.05 < 0.05 Comments: Testicle weight, epididymis, seminal vesicles, prostate gland, levator ani muscles of the groups decreased significantly compared to the group (p 0.05) 3.2.4 Effects of Y10 on histopathology of mice’s testes A Histopathology of mice’s testes (mouse 08, contol group) HE, x 400 C Histopathology of B Histopathology of mice’s testes (mouse 15, group 2) HE,x 400 mice’s testes (mouse 24, group 3) HE, x 400 D Histopathology of mice’s testes (mouse 36, group 4) HE, x 400 E Histopathology of mice’s testes (mouse 42, group 5) HE, x 400 Photo 3.8 Histopathological picture of mouse testicle histogram (HE x 400) Comments: In: The size and image of the sperm tubes of the two groups using Y10 (Groups and 5) and the control group were similar to the group and the tissue space was not much thicker than the physiological group Table 3.17 Diameter of spermatophores of the study groups Groups Group Group Group Group Group (1) (2) (3) (4) (5) Diameter of spermatophores (µm) 128,56 ± 8,60 116,81 ± 9,95 125,95 ± 10,18 125,86 ± 9,88 126,83 ± 10,21 p p1,3,4,5-2< 0,05 p3,4,5-1> 0,05 p4,5-3> 0,05 p4-5> 0,05 Comments: Y10 repaired testicular histopathological lesions, which significantly increased the diameter of the spermatic tubes compared to the non-medication-infected group Y10 at two dosage levels (280 and 560 mg / kg / 24h) had the effect of restoring the diameter of the sperm tube to the equivalent of group (p> 0.05) 3.3 Results of assessing the safety and increasing the ability of sperm stimulation of Y10 in patients with sperm decline 3.3.1 Result of serum testosterone, LH, and FSH concentrations Table 3.18 Serum concentration of testosterone, LH, FSH Indexes LH (IU/l) FSH (IU/l) Testosteron (nmol/l) Before the treatment x After the treatment x Pbefore-after ± SD 6.02 ± 2.14 8.16 ± 4.01 ± SD 5.08 ± 2.06 6.85 ± 3.69 < 0.05 < 0.05 14.65 ± 6.27 16.89 ± 6.42 < 0.05 Comments: serum LH and FSH levels after the treatment decreased compared to before the treatment, the serum testosterone levels after the treatment increased compared to before the treatment The LH and serum FSH of the patients with hormonal disorders increased while the serum testosterone reduced After the treatment, LH and serum FSH decreased, the increase in the serum testosterone was a positive change, the concentration of sex hormones returned to the normal physiological limits 3.3.2 Results of semen in patient Table 3.19 Ratio of semen samples according to sperm classification Number of semen samples Low sperm count Weak sperms Abnormal sperms Low sperm count and weak Weak and abnormal sperms Low sperm count weak and abnormal Normal seminogram results Total Before the treatment After the treatment Pbeforeafter n % n % 9 20.00 30.00 30.00 3.33 8 23.33 26.67 26.67 > 0.05 > 0.05 6.67 3.33 < 0.05 10.00 3.33 16.67 > 0.05 > 0.05 Comments: The results from Table 3.31 show that the percentage of semen samples with low sperm count, weak sperm and deformity decreased after the treatment (p 0.05 6.79 ± 1.26 6.34 ± 1.49 < 0.05 Comments: The semen volume after thettreatment increased, the number of white blood cells after the treatment decreased significantly (p 0.05 No chromosomal structural disorders and chromosomal cluster disorders were found The effects of improving fertility of Y10 - On experimental animals The male white rats that caused sperm deficiency with Natri valproate, Y10 224 mg of medicinal herbs / kg / day and 448 mg of medicinal herbs / kg / day in weeks had the same indicators (serum testosterol concentration, sperm quantity and quality, weight of sexual organs and testicular histology) as the control group This effect is equivalent to the testosteron 16mg / kg / day - On patients with sperm decline - Y10 capsules / day, in months, is safe and effective in treating patients with sperm decline - increases the concentration of serum testosterone, regulates the secretion of LH and FSH, and increases the sperm count and quality - The ‘very good clinical treatment results’ (the semen increases to normal) is 16.67%; Good (increase in sperm count and quality c) is 66.67%; The average (increases the either the sperm count or quality) is16.67%; No result is 0% - improves symptoms of the disease according to traditional medicine - does not cause any undesirable effects; does not alter blood biochemical indices; does not change the hematological index RECOMMENDATIONS It is possible to conduct more research to assess the effect of the medication on other types of sperm decline according to traditional medicine, as well as other pharmacological effects such as antioxidant, liver function protection, enhancement immune system, cardiovascular effects, etc to prove the therapeutic of Y10 It is advisable to massively produce the medication due to its effects proven in the study  ... 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