http://blog.sina.com.cn/bb13 Study on the improvement of human sperm acrosome enzyme activity by Qiang Jing Decoction Bin1 Wang J1 Wang Qiansheng1 Li Guofeng2 (1. Department of Gynecology, The First Affiliated Hospital of Guangxi College of Traditional Chinese Medicine, Nanning 530023 Bin Bin, Department of Gynecology, The First Affiliated Hospital of Guangxi College of Traditional Chinese Medicine 2. Department of Gynecology, Yangjiang Traditional Chinese Medicine Hospital, Yangjiang 529500, Guangdong Province, China) Abstract. OBJECTIVE: To observe the effect of Chinese herbal medicine Qiang Jing Decoction on human sperm acrosome enzyme activity. METHODS: Sixty patients diagnosed with low acrosome enzyme activity and oligozoospermia were divided into two groups of 30 cases each by random number table method. The treatment group was given orally the strong sperm decoction, 1 dose daily, morning and evening, and the control group was given orally the spermogenic capsule, 4 capsules/time, 3 times/day. The treatment cycle was three months for both groups. Both groups measured acrosomal enzyme activity by modified Kennedy method before and after treatment, and measured sperm dynamic quality by Weili Color Sperm Quality Testing System before, at the end of 4 weeks, at the end of 8 weeks, and after treatment. The obtained data were processed by SPSS 11.5 statistical package. RESULTS: 1. Both Strong essence decoction and spermogenic capsule improved the acrosome enzyme activity and the density and vitality of semen of the patients, and the difference between before and after was significant; 2. The effect of strong essence decoction in enhancing sperm vitality was similar to that of spermogenic capsule, and the difference between the two groups was not significant; 3. The effect of strong essence decoction in improving the acrosome enzyme activity of sperm was better than that of spermogenic capsule, and the difference between the two groups was significant. Conclusion: The Chinese herbal medicine compound Qiang Jing Decoction, which mainly tonifies the kidney and strengthens the spleen, and also clears heat, relieves dampness and activates blood circulation to dispel blood stasis legislation, has therapeutic effects on male infertility caused by decreased sperm acrosome enzyme activity in humans. Keywords: strong semen decoction; male infertility; acrosome enzyme activity; Chinese medicine therapy Sperm acrosome enzyme acts as a lysis zona pellucida when sperm penetrate the zona pellucida of oocytes, thus its activity level can directly affect conception [1~3]. The acrosome enzyme activity test is an important indicator of the quality of live sperm [4-5]. Based on our long-term clinical experience, we concluded that “deficiency of both spleen and kidney, combined with dampness, heat, stasis and toxicity” [?] [?] The experimental formula Qiang Jing Decoction [6], which is guided by this theory, is effective in tonifying the kidney and strengthening the spleen, as well as invigorating blood, clearing heat, and relieving dampness, etc. Studies have shown that this formula has the effect of improving sperm quality, repairing testicular pathological damage in model mice [7], and reducing the apoptosis rate of spermatogenic cells [8]. In this study, the effect of Qiang Jing Decoction on human sperm acrosome enzyme activity was observed in a controlled manner to provide a human experimental basis for the study of this type of formula in the treatment of male infertility. 1 Materials and methods 1.1 Materials 1.1.1 Sources of medical records and diagnostic and exclusion criteria 1.1.1.1 Sources of medical records All 60 cases were obtained from patients attending the male outpatient clinic of the First Affiliated Hospital of Guangxi College of Traditional Chinese Medicine from February 2009 to December 2009. 1.1.1.2 Diagnostic criteria: ① Oligozoospermia: refer to the diagnostic criteria recommended by the WHO Laboratory Test Manual for Human Semen and Sperm-Cervical Mucus Interaction [9] in 2001. That is, sperm density <20×106/ml, or total sperm count <40×106 is oligospermia; the number of fast forward-moving sperm (class a) + slow or sluggish forward-moving sperm (class b) <50%, or class a <25%, or class a + class b + non-forward-moving sperm (class c) <60% within 60 min after ejaculation is weak spermatozoa; ② acrosome hypofunction: sperm acrosome (②) acrosome hypofunction: spermatozoa with acrosome activity lower than the lower limit of the normal value recommended by the kit (normal value: 48.2 ~ 218.7 μIU/106 sperm) are considered as acrosome hypofunction; (③) married men aged 22 ~ 45 years old, with infertility duration of 1 ~ 10 years. 1.1.1.3 Inclusion criteria: three of the diagnostic criteria were met. 1.1.1.4 Exclusion criteria: The following conditions were excluded from this observation: chromosomal abnormalities causing infertility; serious reproductive tract infections; combined liver and kidney function and cardiovascular and cerebrovascular abnormalities requiring long-term medication; patients with neurosis and psychiatric disorders; patients with varicocele II or above; patients with sperm density <5×106/ml. 1.1.1.5 Withdrawal criteria: The safety and compliance of medication should be recorded at biweekly follow-up visits, and the trial should be withdrawn if there are two missed visits, or if the medication compliance is <80%, or if the treatment alone is not suitable due to serious adverse events; the trial should also be withdrawn if the spouse is pregnant during the treatment period. Those who withdrew kept their medical records, and their last test result was taken as the final result. 1.1.2 Drugs Strong essence decoction: using single herbal formula granules, produced by Jiangsu Jiangyin Tianjiang Pharmaceutical Co., Ltd. Its main drug composition and dosage (equivalent to the amount of drinking tablets): Cuscuta sinensis 20g, Fructus Lycii 20g, Yimoucao 30g, Deer horn cream 10g, Astragalus membranaceus 30g, Radix Angelicae Sinensis 10g, Conjugate 10g, Oyster 30g, Shenquan 10g, etc. Raw essence capsule: produced by Zunyi Liao Yuanhe Tang Pharmaceutical Co., Ltd, batch number: 20080701. specification: 24 capsules/box, 0.4g/capsule. 1.2 Experimental method 1.2.1 Patient grouping The patients were divided into the control group taking Shengjing capsule and the treatment group taking Qiangjing decoction by random number table method, 30 cases in each group. 1.2.2 Drug administration method and specimen collection ①Drug administration method: The control group was given raw essence capsule orally, 4 capsules/time, 3 times/day; the treatment group was given strong essence decoction, 1 dose/day, the drugs were poured into a cup, added about 200 ml of hot boiling water, stirred to melt, if there was a clot, it could be left to soak for about 10 minutes and then continue to stir, both divided into 2 portions, 1 portion each in the morning and evening. Both groups were treated continuously for 12 weeks. Other drugs used for the treatment of this disease were stopped 2 weeks before and during the treatment period, and were suspended during the period of cold and fever; if other diseases needed to be treated, drugs that had no effect on this disease were used. ②Specimen collection: Sperm was collected by masturbation method before the start of treatment and within 3 days at the end of the 4th, 8th and 12th week of treatment, respectively, with abstinence for 3-7 days before sperm collection, and the collected semen was sent for examination immediately. 1.2.3 Testing of samples ①Sperm dynamics: testing was performed with the Weili Color Sperm Quality Testing System (WLJY-9000 type). ②Sperm acrosome enzyme activity: detected by modified Kennedy method. 1.2.4 Criteria for determining efficacy: Refer to the WHO Standard Examination and Diagnosis Manual for Infertile Couples [10] and the relevant efficacy criteria in the Guidelines for Clinical Research on New Chinese Medicines [11] of the Ministry of Health combined with the actual clinical situation for revision. Cured: spouse conception or sperm acrosome enzymes, density and motility were restored to normal; Effective: although the sperm routine test was abnormal, the sperm acrosome enzymes, motility and a-grade or a+b-grade motile sperm were elevated ≥ 30%, and the sperm density was elevated ≥ 2×106/ml after treatment of oligospermia; Ineffective: those who did not reach the above criteria after treatment. 1.2.6 Statistical processing method All experimental data were processed by SPSS11.5 statistical package, and the difference was considered significant at P < 0.05 and highly significant at P < 0.01. 2 Experimental results 2.1 Symptoms, signs and general observations All the patients in the two groups of this experiment took all the drugs as required, and no significant adverse reactions were observed. 2.2 Semen test results The data of acrosome enzyme activity, semen volume, sperm density, sperm vitality, sperm viability, liquefaction time and pH value in the test conducted before treatment in the two groups were tested by t-test, P > 0.05, and the difference was not significant. After treatment, the acrosome enzyme activity, sperm density, sperm motility and viability of the two groups were compared with those before treatment in the same group by t-test, P < 0.05, and the difference was significant. There was a significant difference in the enhancement of acrosome enzyme activity in the strong semen decoction group compared with the spermogenic capsule group by t-test, P < 0.05. The difference between the two groups in terms of semen volume, liquefaction time and pH before and after treatment was not significant by t-test, P > 0.05. Comparison of sperm density, sperm vitality and viability between the two groups before and after treatment, by t-test, P > 0.05, the difference was not significant. See Table 1: Table 1 Comparison of semen specimen testing before and after treatment (±s) Group Number of cases Acrosome enzyme activity (μIU/106 sperm) Semen volume (ml) Sperm density (×106/ml) Strong sperm decoction group Before treatment After treatment Before treatment After treatment 30 30 30 30 30 29.879±10.513△ 49.548±8.443*◆ 31.686±8.748 42.797±9.000* 3.54±0.81△ 3.61±0.62 3.68±0.77 3.57±0.51 18.31±7.34△ 28.15±6.25* 18.87±6.70 27.46±6.19* Spermatin capsule group Note: △ P > 0.05 by t-test before treatment compared with spermatin capsule group; *Spermatin capsule group compared with the same group after treatment by t-test. P < 0.05 by t-test after treatment compared with before treatment; ◆P < 0.05 by t-test compared with the sperm production capsule group after treatment. Table 1 (continued) Comparison of semen specimens tested before and after treatment (±s) Group Number of cases Sperm viability class a (%) Sperm viability class a+b (%) Sperm viability class a+b+c (%) Strong sperm decoction group Sperm production capsule group Before treatment After treatment Before treatment After treatment 30 30 30 30 30 8.16±4.57△ 23.53±5.72* 6.08±4.28 22.52±4.10* 15.90±6.11△ 47.63±10.36* 15.67±4.88 45.34±7.56* 30.29±8.00△ 64.04±12.40* 30.82±7.82 60.48± 6.81* Table 1 (continued) Comparison of semen specimen testing before and after treatment (±s) Group Number of cases Liquefaction time (min) pH value Strong semen decoction group Raw semen capsule group Before treatment After treatment Before treatment After treatment 30 30 30 30 30 22.0±5.54△ 21.57±3.91 22.0±5.42 22.57±5.96 7.48±0.28△ 7.48± 0.23 7.45±0.34 7.38±0.34