• The Journal of Korean Medicine
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• v.26 no.4
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• pp.12-21
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• 2005

Objectives: In the present study, we investigated whether an aqueous extract of Armeniacae semen induces apoptotic neuronal cell death upon mouse N2a neuroblastoma cells. Methods: 1. Cell viability was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTI) assay. 2. For in situ detection of apoptotic cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining. 3. The fraction of cells was revealed by flow cytometric analysis used that. 4. For detection of apoptotic DNA cleavage, DNA fragmentation assay was performed. 5. For detection of bax and bcl-2, Western blot analysis was performed. 6. Caspase enzyme activity was measured using caspase-3 assay. Results: From the present results, N2a neuroblastoma cells treated with Armeniacae semen extract exhibited several characteristics of apoptosis. A treatment of Armeniacae semen extract was shown to increase the expression of Bax, a proapoptotic protein, and the treatment decreased the expression of Blc2, an anti-apoptotic protein. In addition, Armeniacae semen extract increased the caspase-3 enzyme activity. Conclusions: The present results show that Armeniacae semen extract induces apoptotic cell death in mouse N2a neuroblastoma cells.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.311-318
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• 1996

Mycoplasmas have long been suspected of contributing to involuntary infertility in couples. However considerable disagreement exits concerning the role of genital mycoplasma infection in human infertility. Several investigators have noted abnormalities in the semen analysis of men with positive mycoplasma cultures, and early epidemiologic studies indicated that Ureaplasma urealyticum was linked to human reproductive failure on the basis of higher frequencies of isolation from infertile versus fertile couples and successful pregnancies in infertile couples after doxycycline therapy. However, subsequent investigators have questioned these findings because there are many studies in which treatment for mycoplasma in the male or female did not demonstrate an improved pregnancy rate, and semen samples from unexplained infertile men containing ureaplasmas have not revealed poorer motility, fewer spermatozoa and more aberrant forms. The objective of this study were to investigate the incidence rate of mycoplasma in semen and to investigate whether the presence of mycoplasma in semen makes significant difference to the semen volume, sperm motility and sperm counts. The results were that the rate of isolation of mycoplasma species was 70.3%. Semen volume is $2.84{\pm}1.01ml$ for culture negative and $3.15{\pm}1.42ml$ for culture positive group. Sperm motility is $46.23{\pm}15.80%$ for culture negative and $50.09{\pm}15.69%$ for culture positive group, and sperm count is $95.47{\pm}47.14({\times}(P)10^6/ml)$ for culture negative and $86.73{\pm}47.59({\times}10^6/ml)$ for culture positive group. In conclusion, we suggest that the presence of mycoplasma in semen makes no significant differences to the sperm parameters.

• Korean Journal of Pharmacognosy
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• v.32 no.2 s.125
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• pp.157-162
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• 2001

For the quality control of Cassiae Semen, anthraquinone compound, aurantio-obtusin, was isolated from the MeOH extract of Cassiae Semen (Leguminosae) and identified by the spectroscopic analysis. A quantitative analysis of aurantio-obtusin using HPLC method showed that the average contents of aurantio-obtusin were $0.03{\pm}0.01%$ in 50 samples collected throughout the various regions of Korea.

• Archives of Pharmacal Research
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• v.26 no.2
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• pp.157-161
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• 2003

The major ingredient of Persicae Semen is a cynogenic compound, amygdalin (D-mandelonitrile-$\beta$-gentiobioside). Controversial results on the anticancer activity of amygdalin were reported due to its conversion to its inactive isomer, neoamygdalin. In order to inhibit the epimerization of amygdalin, we used newly developed simple acid boiling method in preparation of Persicae Semen extract. HPLC analysis revealed most of amygdalin in Persicae Semen extract was active D-form. Persicae Semen extract was used to analyze its effect on cell proliferation and induction of apoptosis in human promyelocytic leukemia (HL-60) cells. Persicae Semen extract was cytotoxic to HL-60 cells with $IC_{50}$ of 6.4 mg/mL in the presence of 250 nM of $\beta$-glucosidase. The antiproliferative effects of Persicae Semen extract appear to be attributable to its induction of apoptotic cell death, as Persicae Semen extract induced nuclear morphology changes and internucleosomal DNA fragmentation.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.2
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• pp.142-149
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• 2021

Objective: Bacteriospermia and urogenital infections are common problems in male infertility. This study aimed to evaluate the effects of bacteriospermia on sperm parameters and clinical outcomes in semen samples infected with two common bacteria (Staphylococcus saprophyticus and Escherichia coli) in northern Iran. Methods: Microbiological tests were performed to isolate and identify organisms from 435 semen samples from infertile couples. Semen samples were assessed according to the World Health Organization criteria. The protamine status, chromatin structure, chromatin condensation, and acrosome reaction of sperm and assisted reproductive outcomes were determined in couples with different male infertility factors. Results: Among the total cases, the two most prevalent pathogens were considered: S. saprophyticus (38.2%) and E. coli (52.9%). In the semen samples infected with E. coli, the spontaneous acrosome reaction and abnormal chromatin condensation were more common (p<0.05). Significant increases in abnormal chromatin condensation and deprotamination were seen in the presence of S. saprophyticus. In washed semen, tight adhesion between the sperm midpiece and S. saprophyticus was observed. There was also a significant decrease in the fertilization rate using semen samples infected with S. saprophyticus and E. coli during in vitro fertilization cycles (p<0.001). In addition, the presence of S. saprophyticus and E. coli in semen samples was associated with a lower likelihood of clinical pregnancy in couples with various factors of male infertility. Conclusion: Poor results of assisted reproductive techniques may be correlated with semen samples infected with two common bacteria in northern Iran.

• Clinical and Experimental Reproductive Medicine
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• v.3 no.2
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• pp.35-42
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• 1976

The human semen ejaculated in a form of liquid state, coagulates immediately after ejaculation, and then liquefies again. However, the mechanisms of neither coagulation and liquefaction of semen have not been explained clearly so far, and very limited numbers of report are available, although the spermatology and andrology made rapid progress. This clinical study has been undertaken to investigate the liquefaction phenomena and practicability of the results might be applied to fertility and infertility problems. As a preliminary study, in this report the liquefaction time of various semen groups is measured and analysed. The following results are obtained: 1. An average liquefaction time of semen of a total of 60 subjects: 25 minutes. 2. An average liquefaction time of semen according to sperm count: 1) Normospermia group (20 cases): 34 minutes. 2) Oligospermia group (20 cases): 21 minutes. 3) Azoospermia group (20 cases): 20 minutes. 3. An average liquefaction time of semen according to abstinence period: 1) Less than 3 days group (30 cases): 22 minutes. 2) More then 5 days group (30 cases): 28 minutes. In conclusion: 1. The liquefaction time of semen of the normospermia group is longer than oligospermia group or azoosermia group. 2. The liquefaction time of semen may not be greatly influenced by the various factors such as abstinence period, semen volume, semen pH, age of the subjects and so on. 3. In routine semen analyses, it is recommended to begin the analysis at least 25 minutes after the ejaculation. 4. Further studies are required in conjunction with practical application of liquefaction mechanism in infertility and fertility control.

• Korean Journal of Pharmacognosy
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• v.32 no.1 s.124
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• pp.39-42
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• 2001

The seeds of Areca catechu L. has been used for the treatment of the diseases caused by parasites in East Asia. As a part of a research for standardization of crude drugs, we have determined the content of arecoline in the seeds of Arecae Semen purchased from various regions of Korea. The HPLC method for quantitative analysis of arecoline in Arecae Semen was established and reproducible results and chromatographic isolation of arecoline was accomplished successively. It suggested that the content of arecoline in Arecae Semen was $0.2726\;{\pm}\;0.05532%$.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.3
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• pp.196-201
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• 2022

Objective: This prospective consecutive study investigated the variation in sperm DNA fragmentation (SDF) in multiple semen samples from patients with cancer. Methods: Eighty-one patients with various cancers underwent multiple semen collections on 3 consecutive days for sperm cryopreservation prior to cancer treatment. A commercial Halosperm kit was used to measure SDF. Within- and between-subject coefficients of variation were estimated via random-effects analysis of variance to assess the consistency of semen parameters and SDF. Intraclass correlation coefficients (ICCs) were calculated to assess the magnitude of the between-subject component of variance relative to the total variance. Results: The volume of semen in the day-2 and day-3 samples was significantly lower compared with the day-1 sample. Most parameters showed high ICC values, suggesting that within-subject fluctuations were small relative to the between-subject variability. The highest ICC values were identified for the SDF (ICC, 0.68; 95% confidence interval [CI], 0.45-0.84) and semen volume (ICC, 0.67; 95% CI, 0.45-0.84). Conclusion: Our findings showed that repeated ejaculates from patients with cancer had stable SDF levels.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.965-972
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• 2005

To investigate the effects of Trichosanthes semen extract on the growth and apoptosis of human uterine cervical carcinoma cells. Effects of Trichosanthes semen extract on the growth of ME-180 cells were assayed by MTT assay. Apoptosis induced by Trichosanthes semen extract was detected by fluorescent microscopy, DNA fragmentation analysis and flow cytometry. Caspase-3 and caspase-8 activities were assayed. Trichosanthes semen extract induced ME-180 cells to die in a dose- and time-dependent manner. ME-180 cells treated with Trichosanthes semen extract exhibited typical characteristics of apoptosis. The population of Sub-G1 cells increased significantly, and the cells represented the reduced size, condensed chromatin and apoptotic bodies. They showed the decreased mitochondrial membrane potential and increased activities of caspase-3 and caspase-8. The results suggest that Trichosanthes semen extract induced ME-180 cell apoptosis and the activation of caspase and mitochondrial pathway were involved in the process of Trichosanthes semen extract-induced apoptosis.

• Korean Journal of Pharmacognosy
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• v.40 no.1
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• pp.18-24
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• 2009

GCSB-5 preparation is a purified extract from a mixture six herbal medicines (Acanthopanacis Cortex, Achyranthis Radix, Saposhnikoviae Radix, Cibotii Rhizoma, Glycine Semen Nigra, Eucommiae Cortex) that have been widely used in traditional medicine to treat various bone disorders. This study was carried out to obtain the HPLC analysis method that can be used to establish quantitative analysis of Glycine Semen Nigra and Eucommiae Cortex for standardization of GCSB-5 preparation. HPLC analysis methods for the simultaneous determination of genistin (Glycine Semen Nigra) and geniposide (Eucommiae Cortex) were established for the quality control of herbal medicinal raw material and preparation. And validation of HPLC analysis methods were conformed for verification of HPLC methods by check to specificity, linearity, intra-day precision, inter-day precision and accuracy following ICH guideline. As the result of quantitative analysis, the contents of genistin and geniposide in the raw material of GCSB-5 preparation were 0.0426-0.0427 mg/g and 0.431-0.432 mg/g. And GCSB-5 preparation contained genistin of 0.0202-0.0203 mg/capsule and geniposide of 0.211-0.212 mg/capsule, respectively.