Objectives To check marker content for appropriate quality control of Woohwangchungsimwon sterilized to ensure microbiological safety and to observe antioxidant activity for any changes in efficacy. Methods To measure any effects of sterilization on the effective compounds, 8 ingredients of Woohwangchungsimwon were screened for any changes in marker content using HPLC-DAD. Using the colorimetric method on the microplate reader any changes in total phenolic compound and flavonoid levels were observed. Antioxidant activity was measured using the DPPH, ABTS, and FRAP. Results Of the ingredients of Woohwangchungsimwon, 8 were subject to quantitative analysis before and after sterilization. 21.6 mg and 1.93 mg of Glycyrrhizin was found in Glycyrrhiza uralensis Fischer pre and post sterilization, respectively. Decursin found in Angelica gigas Nakai increased from 0.16 mg to 0.29 mg after sterilization. Bilirubin found in Gallstone of Bostaurusvar. domesticus increased from 0.24 mg to 0.33 mg. Cinnamic acid found in Cinnamomum cassia Blume increased from 0.02 mg to 0.05 mg. Ginsenoside Rb1 found in Panax ginseng C. A. Meyer decreased from 0.02 mg to 0.14 mg. Paeoniflorin found in Paeonia lactiflora Pallas increased from 1.05 mg to 1.13 mg. Amygdalin found in Armeniacae Amarum Semen increased from 2.68 mg to 2.83 mg. L-muscone found in Musk increased from 0.63 mg to 0.76 mg. As for total phenolic compound and total flavonoid content, there was a 1.22 and 4.15-fold increase. DPPH and ABTS increased by 20.45% and 20.69%, respectively. FRAP activity was 2.78 times more active post stabilization. Conclusions This study confirmed that high temperature high pressure steam sterilization, a method used to ensure microbiological safety of Woohwangchungsimwon, does not affect marker content; in other words, does not affect quality of the Woohwangchungsimwon. It could also be seen that total phenolic compound and flavonoid content increased after sterilization. An antioxidant activity test showed that there was significantly increased activity of antioxidants.
Objective: The Shanghai Holstein cattle breed is susceptible to severe mastitis and other diseases due to the hot weather and long-term humidity in Shanghai, which is the main distribution centre for providing Holstein semen to various farms throughout China. Our objective was to determine the genetic mechanisms influencing economically important traits, especially diseases that have huge impact on the yield and quality of milk as well as reproduction. Methods: In our study, we detected the structural variations of 1,092 Shanghai Holstein cows by using next-generation sequencing. We used the DELLY software to identify deletions and insertions, cn.MOPS to identify copy-number variants (CNVs). Furthermore, we annotated these structural variations using different bioinformatics tools, such as gene ontology, cattle quantitative trait locus (QTL) database and ingenuity pathway analysis (IPA). Results: The average number of high-quality reads was 3,046,279. After filtering, a total of 16,831 deletions, 12,735 insertions and 490 CNVs were identified. The annotation results showed that these mapped genes were significantly enriched for specific biological functions, such as disease and reproduction. In addition, the enrichment results based on the cattle QTL database showed that the number of variants related to milk and reproduction was higher than the number of variants related to other traits. IPA core analysis found that the structural variations were related to reproduction, lipid metabolism, and inflammation. According to the functional analysis, structural variations were important factors affecting the variation of different traits in Shanghai Holstein cattle. Our results provide meaningful information about structural variations, which may be useful in future assessments of the associations between variations and important phenotypes in Shanghai Holstein cattle. Conclusion: Structural variations identified in this study were extremely different from those of previous studies. Many structural variations were found to be associated with mastitis and reproductive system diseases; these results are in accordance with the characteristics of the environment that Shanghai Holstein cattle experience.
Kim, Jeong A;Cho, Eun Seok;Jeong, Yong Dae;Choi, Yo Han;Hong, Jun Ki;Kim, Young Sin;Chung, Hak Jae;Baek, Sun Young;Sa, Soo Jin
Journal of the Korea Academia-Industrial cooperation Society
/
v.21
no.9
/
pp.251-258
/
2020
This study investigated the effect of Zardaverine supplementation in freezing extender, on kinetic characteristics of post-thawed boar sperm. Cryopreservation of boar sperm is an important technique of assisted reproductive technology and genetic resource banking. Although this technique is particularly useful, freeze-thaw cycles associated with sperm cryopreservation significantly reduce sperm quality. Semen from mature Duroc boars were collected and cryopreserved in freezing extenders (LEY) treated with varying concentrations of Zardaverine (0, 20, 50, 75, 100 𝜇M). The time-dependent kinetic characteristics of post-thawed spermatozoa were determined after thawing by applying computer-assisted sperm analysis (CASA). We observed that the motility immediately after thawing was significantly higher in 20 𝜇M stocks than in control (0 𝜇M) and the other treatments (p<0.05). Curvilinear velocity (VCL) in 0 𝜇M and 20 𝜇M stocks were significantly higher than the other treatment groups, except 75 𝜇M (p<0.05). Higher average path velocity (VAP) was obtained at 20 𝜇M as compared to 100 𝜇M, whereas amplitude of head lateral displacement (ALH) was significantly higher at 20 𝜇M than 50 𝜇M and 100 𝜇M (p<0.05). No differences were obtained for Straight-line velocity (VSL) and Linearity (LIN). In conclusion, our results indicate that Zardaverine improves the motility, VCL, VAP, and ALH of post-thawed boar sperm.
Objectives: This review investigated the research on warm herbal foot bath therapy (WHFT) for insomnia. Methods: A search was conducted on the China National Knowledge Infrastructure (CNKI) database to collect relevant studies published up to August 29, 2023. Randomized controlled trials (RCTs) comparing WHFT and sleeping pills in patients with insomnia were included. The methodological quality of the included studies was assessed using the Cochrane risk-of-bias assessment tool. The results of the meta-analysis were presented as risk ratios (RRs) or mean differences (MDs) and their 95% confidence intervals (CIs). Results: A total of 11 RCTs were included. WHFT as monotherapy resulted in a significantly higher total effective rate (TER) (RR, 1.25; 95% CI, 1.15 to 1.36; I2=25%) and an improved Pittsburgh Sleep Quality Index (PSQI) global sore (MD, -3.10; 95% CI, -4.24 to -1.95; I2=73%) compared to benzodiazepines. Additionally, WHFT as a combined therapy with benzodiazepines resulted in a significantly higher TER (RR, 1.15; 95% CI, 1.04 to 1.27; I2=0%) and an improved PSQI global score (MD, -2.23; 95% CI, -4.09 to -0.38; I2=80%) compared to benzodiazepines alone. In network analysis visualizing the components of HWFT, four clusters were discovered, and Polygoni Multiflori Ramuls and Ziziphi Spinosae Semen were the key herbs used in WHFT. Overall, the methodological quality of the included studies was poor. Conclusions: There was limited evidence that WHFT as a monotherapy or combined therapy was effective in improving insomnia. The findings can be used as basic data for future WHFT research in South Korea.
Kim, Seok-Hyun;Pang, Myung-Geol;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chang, Yoon-Seok
Clinical and Experimental Reproductive Medicine
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v.18
no.1
/
pp.63-71
/
1991
To establish the normal fertile range in the results of the sperm zona-free hamster ova penetration assay (SPA) in Korean male, SPA using the low temperature ($4^{\circ}C$) capacitation in TEST-yolk buffer (TYB) was performed in 67 fertile and 26 infertile men. Sperm parameters in routine semen analysis were also checked and compared with the results of SPA. Sperm concentration, motility and motility index (MI) were significantly higher in fertile group compared with infertile group: $96.0{\pm}46.6$ vs $43.6{\pm}31.9{\times}10^6/ml$, $65.5{\pm}14.8%$ vs $45.8{\pm}23.6%$ and $46.31{\pm}13.29$ vs 27.40{\pm}17.98$, respectively. In fertile group, the hamster ova penetration rate (PR) was $98.5{\pm}5.0%$ (80%-100%), and the penetration index (mean penetrations per ovum, PI) was $9.59{\pm}6.35$(3.1-29.0). All the fertile men showed PI>3.0. In infertile group, PR was $24.6{\pm}24.8%$ (0%-70%), and PI was $0.40{\pm}0.42$ (0-1.3). Both PR and PI were significantly lower in infertile group. There was a significant correlation beween PI and sperm motility or MI, respectively, in fertile group whereas there was no correlation in infertile group. These data suggest that SPA using the low temperature capacitation in TYB can be a valuable diagnostic tool for the assessment of male fertility in vitro and provide an important supplement to the traditional tests of sperm quality.
This study was to investigate pregnancy rate of IVM/IVF/IVC Korean cattle (registered in government) embryos according to transport time course. For the production of embryos, oocytes recovered from slaughtered excellent grade cow and highly motile frozen-thawed bull semen (purchased from LIMC, KPN#497) was used. In vitro produced embryos were cultured in CR1aa medium for 8 days and some of them were frozen. The rate of average cleavage (>2-cell) was 83.0% (308/371) and blastocyst rate at day 8 was 34.7% (107/308). Among in vitro produced blastocyst embryos at day 8, most healthy embryos were freshly transferred on production day and some frozen embryos were direct transferred on appropriate day. These embryos were produced in a laboratory, embryo transfer (ET) was planned in 10 areas of the remote island (Jeju) from the laboratory by airplane. Thus, we examined the pregnancy rate in recipient cow according to embryo of transport time course before ET. From embryo transferred 44 recipient cows, overall pregnancy was 40.9% (18/44), these 18 cows were all calved [single, 94% (17/18); twin, 6% (1/18)] and total embryo implantation rate was 26% (19/66). Comparing transport time in the base of 6 hr, pregnancy rate in ET group required less 4 hr (60%, 9/15) was significantly higher than that required more 6 hr (26.3%, 5/19). In direct ET of freezing embryos, the pregnancy rate was 40% (4/10). However, it was difficult to find the meaning of temperature, pH and corpus luteum quality of recipients on comparison of pregnancy rate. When the cell death level of embryos according to storage time in thermos (straw container) before ET was measured by TUNEL staining, apoptotic index was increased with storage time-dependent. These results demonstrated that long distance transfer of IVM/IVF/IVC embryos is possible and the time of embryo transport is very important for the pregnancy rate on field trial.
Cryopreservation is commonly used for an efficient utilization of semen, oocytes and embryos but has disadvantage in the survival, development of the post-thawed eggs. The high risk in the survival, development of eggs after thawing is thought to be caused by inappropriate internal regulation of $Ca^{2+}$ and/or formation of intracellular ice crystals. In this experiment, we tested whether the $Ca^{2+}$ current (iCa), a decisive factor to $Ca^{2+}$ entry, was altered in post-thawed oocytes by using whole cell voltage clamp technique. The quality and survival rates of the oocytes derived from both fresh and frozen groups were examined by morphology and FDA-test. Vitrified oocytes (VOs) were incubated for 4 hr after thawing and then donated to this experiment. Ethyleneglycol-ficoll-galactose (EFG) was used as a cryoprotectant for vitrification. The membrane potential was held at -80 mV and step depolarizations of 250 ms were applied from -50 mV to 50 mV in 10 mV increments. The survival rates showed a higher in VOs vitrified with EFG containing $Ca^{2+}$ than in VOs vitrified with EFG under the $Ca^{2+}$-free condition (82.0% vs 14%). In group with/without $Ca^{2+}$, the survival rates were significantly (P<0.01) difference. In the fresh metaphase II oocytes (FOs), current-voltage (I-V) relationship showed that iCa began to activate at -40 mV and reached its maximum at -10 mV. With same voltage pulses, inward currents were elicited in VOs. I-V relationships observed in VOs were similar to those in FOs. Time constants of activation and inactivation of the inward current shown in VOs were not different to those in FOs. This accordance in I-V relations and time constants in FOs with those in VOs indicates that the inward currents in FOs are unaltered by vitrification and thawing. Therefore, vitrification with EFG does not play as a factor to deteriorate $Ca^{2+}$ entry across the membrane of the oocytes.
Kim, Eun-Ji;Talha, Nabeel A.H.;Jeon, Yu-Byeol;Yu, Il-Jeoung
Journal of Animal Reproduction and Biotechnology
/
v.34
no.1
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pp.57-63
/
2019
This study was conducted to find out the effect that ${\kappa}-Carrageenan$ has on the properties of dog sperm when it was added to the cryoprotectant. Extender basically was contained 1.21 g Trizma base, 0.67 g citric acid, 0.4 g glucose, 0.03 g penicillin G, 0.05 g streptomycin sulfate. Extender1 was added with 0.1%, 0.2%, 0.3%, and 0.5% carrageenan, while extender2 was supplemented with glycerol. After freezing-thawing, the motility, viability, acrosome integrity, apoptosis, and ROS (reactive oxygen specifications) of sperm were measured to analyze the effects of the supplementation of carrageenan. Total Motile (TM), Rapid Progressive Motile (RPM), Medium Progressive Motile (MPM), and Immotile were measured through the CASA system after thawing in 37 degree water. Extender with 0.2% ${\kappa}-carrageenan$ ($64.26{\pm}0.49$) was significantly higher than control ($40.24{\pm}8.27$) (p < 0.05). RPMs of extender with 0.1%, 0.2% ${\kappa}-carrageenan$ ($57.64{\pm}6.34$, $56.47{\pm}1.35$) were significantly higher than the other groups (p < 0.05). Acrosome integrity was measured by dyeing to PSA-FITC with an epifluorescence microscope. Normal acrosome ratio of extender with 0.5% ${\kappa}-carrageenan$ ($61{\pm}8.03$) was higher than the other groups (p < 0.05). Apoptosis was measured with a FACSCalibur flow cytometer using FITC (FITC Annexin V Apoptosis Detection Kit). Treated groups of ${\kappa}-carrageenan$ of 0.1% ($0.81{\pm}0.05$), 0.2% ($0.85{\pm}0.05$) were significantly higer (p < 0.05) than control. Modified SYBR/PI staining was used for determination of viability and DCF staining was used for evaluation of ROS. Viability and ROS were not significantly different from other groups. In conclusion, adding a certain concentration of carrageenan to the extender of cryopreservation, carrageenan contributes to the improvement of the sperm motility, acrosome integrity and prevention of apoptosis.
Journal of Korean Society of Occupational and Environmental Hygiene
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v.31
no.3
/
pp.202-212
/
2021
Objectives: The reproductive health of female workers has been extensively investigated in South Korea and other countries worldwide. However, few studies have discussed the reproductive health of male workers. In this study, we reviewed the recent literature that reports on the effects of occupational exposure on the reproductive health of male workers and the health of their children. Methods: In May 2020 we used the PubMed search engine to search the literature over the last 10 years and chose case-control, cohort, and cross-sectional studies and reviews. We selected epidemiological studies that investigated the association between pre-pregnant occupational exposure and the reproductive health of male workers and the health of their children. We excluded case reports, non-epidemiological studies (animal experiments, cellular-level experiments, and similar articles), and studies that described postnatal occupational exposure. Results: We eventually selected 23 studies. The studies that included exclusively male workers reported that those employed in the agricultural sector or those exposed to pesticides showed lower blood levels of reproductive hormones and a high risk of lympho-hematopoietic system cancer in their children. Male workers exposed to complex organic solvents and organic compounds showed a high risk of poor semen quality, increased time to pregnancy, decreased blood levels of reproductive hormones, and a high risk of lympho-hematopoietic system cancer in their children. Male workers employed in occupations that involved significant social contact, or in the leather and livestock industries, and in occupations with high levels of exposure to lead and organic solvents showed a high risk of malignancies, including lympho-hematopoietic system cancer, neuroblastoma, and central nervous system tumors in their children. Studies that investigated both male and female workers reported that children of male smelters showed a high risk of premature birth, and children of male workers exposed to metals showed a high risk of hypospadias and cryptorchidism. Children of male welders and workers employed in the glass, ceramic, and tile industries showed a high risk of premature birth. Conclusion: The findings of this study will serve as basic data for further research on male workers' reproductive health and provide a scientific basis for the development of strategies to protect the reproductive health of males employed in high-risk occupations. Moreover, the results of this study may provide guidelines to improve the understanding of and knowledge on male workers' reproductive health.
This study was carried out to assess the effect of superovulation response and quality of embryos recovered from donor cows after a single subcutaneous injection of FSH dissolved in polyethylene glycol (PEG). Cows were allocated into control and 3 experimental treatment groups. In control, cows were injected intramuscularly 50 mg FSH twice daily for 4 days. Group 1 were injected subcutaneously with a single dose of 400 mg FSH dissolved in 30% PEG solution. Group 2 were injected subcutaneously with a single dose of 200 mg FSH dissolved in 30% PEG solution. Finally in group 3, cows were injected twice 200 mg FSH dissolved in 30% PEG solution by subcutaneous. Superovulation was initiated by injection of FSH between Day 8 and 14 of the estrus cycle (Day 0, the day of estrus), and followed by injection of 25 mg PGF$_2$$\alpha$ at 48 h after first FSH injection. Cows were then artificially inseminated (AI) with semen twice at 48 and 60 h after PGF$_2$$\alpha$ injection. At 7 days after the second AI, embryos collected non-surgically by flushing the uterine horns and were counted and compared morphologically as being transferable and degenerated among different superovulation treatments. Furthermore, progesterone and estradiol -17 $\beta$ in plasma were measured by radioimmunoassay following different treatments at given days All cows of treated groups were observed heat. but control group was showed 77.8%. Superovulation response was observed as 77.8, 87.5, 88.9, and 100% in control, Groups 1, 2 and 3 The mean number of corpus lutea (CL) detected in Group 1 were 19.6, which was, respectively significantly (P<0.05) higher than those of other groups (11.1, 13.4 and 7.6, respectively). However, there did not differ on the mean number of total embryos recovered and of transferable embryos between control and treated groups.
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