Kang, Jung-A;Ka, Kang-Hyeon;Kim, Jun Young;Yoon, Seong-Tak;Kim, Seong Hwan
Journal of Mushroom
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v.17
no.4
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pp.191-196
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2019
The edible ectomycorrhizal mushroom Amanita javanica is a valuable species protected by forest law in Korea. However, basic characterization data on its use as an important forest resource has been limited. This study was performed to determine mycelia growth characteristics of the domestically isolated Amanita javanica strain NIFoS 1267 on potato dextrose agar media under diverse culture conditions. Physical factors temperature, pH, and light, as well as chemical factors salts, heavy metals, and pesticides were examined for their effects on the growth of the mushroom strain. The mycelia of A. javanica strain exhibited optimal growth when cultured in dark at 30℃ in media with a pH of 5-6. Normal levels of growth were observed in media containing up to 2% saline. At a heavy metal ion content of 50 ppm, mycelial growth was not affected by arsenic ion but was affected by cadmium and lead ions. In the tests performed with two pesticides used in Korean forests, the growth of the mushroom strain was not affected by the presence of abamectin, but was inhibited in media containing acetamiprid, emamectin benzoate, or thiacloprid. These results are expected to facilitate artificial cultivation of A. javanica as a new commercial product.
The objective of this study was to investigate the in vitro / in vivo embryonic development after vitrification of mouse zygotes and the chrom osomal normality of delivered live young after embryo transfer. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glyc이, 30% Ficoll a and 0.3 M sucrose in phosphate buffer saline c containing 10% FBS ) . After mouse zygotes were exposed to EFS40 at 25"C for 30 sec., they were immediately plunged into LN$_2$. Vitrified thawed mouse zygotes were cultured upto bIastocysts in M16 for 4 days. The rates of in vitro development were 71.5% under this condition. Cultured blastocysts were transferred to recipients (3 day of pseudopregnant) on one or both uterus horns (6-8 embryos per a uterus horn). And all recipients were allowed to produce litters. The results obtained in these experiments were summarized as follows: The pregnancy rates and in vivo survival rates, live fetus rates, for vitrified zygotes (80.0, 39.6%) were not significantly difference in those of control zygotes (77.8%, 50.0%). Also, all of live-born young mice were chromosomally normal (n=40). This results suggested that proposed rapid vitrification procedures can be effectively use in 1-cell mouse zygotes cryopreservation.
Kim, Han-Soo;Baik, Seong-Wan;Kim, Inn-Se;Chung, Kyoo-Sub
The Korean Journal of Pain
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v.1
no.2
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pp.192-198
/
1988
Sixty patients, of ASA physical status class I for elective operations in the lower abdomen, perineum, or lower extremities, were studied in a comparative prospective trial to evaluate the efficacy of epidural morphine and ketamine for postoperative analgesia. They were divided into two groups: an epidural morphine sulfate group (EMS group; 30 patients), and an epidural ketamine hydrochloride group (EKH group; 30 patients). Indwelling epidural catheters were placed in the patients' lumber areas (L3-4) and then all patients were anesthetized with thiopental, nitrous oxide, and halothane. After the patients had fully recovered from the anesthesia, the analgesic agents were administered epidurally via the catheter when the patients complained of pain in the postoperative period. The groups were given either 0.1 mg/kg of morphine sulfate or 0.5 mg/kg of ketamine hydrochloride administered in a volume of 10 ml of normal saline. Patients were observed for the onset and duration of postoperative analgesia and for other effects. Total doses were $5.7{\pm}0.6\;mg$ of morphine sulfate in the EMS group and $27.9{\pm}3.3\;mg$ of ketamine hydrochloride in the EKH group. The onset of analgesia was detectable within 35 min.($23.5{\pm}6.3$ min) in 86.7% (26 cases) of the EMS group and within 10 min. ($7.8{\pm}3.7$ min.) in 76.7% (23 cases) of the EKH group. Mean duration of postoperative analgesia was $22.3{\pm}2.1\;hr$. in the EMS group. In the EKH group, the duration of analgesia was shorter and variable, the range of duration was from 2 hr. to 24 hr., Cardiopulmonary changes were statistically insignificant ih both groups. Side effects such as nausea, vomiting, urinary retention, pruritus, dizziness, and headache were observed in EMS group. In the EKH group, there was no discomfort except dizziness (3 cases) and headache (1 case). Epidural ketamine was a safe technique for postoperative analgesia, but because of the variability and relative shortness in the duration of analgesia the use of this technique will require further clinical trials.
The objective of the present study is to assess the contribution of bulk flow to the regulatory mechanism of amniotic fluid volume and its ionic concentration in the membranes surrounding the amniotic fluid. For quantitative assessment, we prepared 4 kinds of artificial amniotic fIuids (isotonic isovolumetric, hypotonic isovolumetric, isotonic hypervolumetric and hypotonic hypervolumetric ones) by replacing 70% of amniotic fluid of pregnant rabbits with water or normal Tyrode solutions. Isoosmotic saline of 0.5 ml volume containing 0.05% Censored and 15 mM/l LiCl was administered initially into amniotic sacs of all subject animals. Samples of amniotic fluid were collected in after 30 and 90 minute intervals; the concentrations of Censored, $Na^+\;and\;Li^+$ were determined and compared. Followings are the results obtained. 1. from isovolumetric and increased Congcord group, we couldn't find significant change in $Li^+\;and\;Na^+$ concentration in isotonic amniotic fluid. However, $Na^+$ concentration increased significantly as well as a striking increase in Censored concentration in hypotonic amniotic fluid. 2. In isovoIumetric and decreased Censored group, the rate of $[Li^+]$ decrement and the rate of $[Na^+]$ increment were much higher in hypotonic amniotic fluid than in isotonic. 3. In hypervolumetric and increased Censored group, the rate of $Na^+$ efflux increased proportionately with the increment of Censored concentration up to 0.98, which was higher than the rate of $Li^+$ efflux in isotonic amniotic fluid. However, the increment of $Na^+$ concentration was rather related with the initial $Na^+$ concentration in hypotonic amniotic fluid, showing inverse relationship. $Li^+$ concentration increased only when there was a marked increase in Censored concentration and approached near a maximum value or 1. 4. For hypervolumetric and decreased Censored group, the observations were identical to isovolumetric and decreased Censored group. From these results the following conclusions could be made: 1) There is no net movement of water or monovalent cations across the membranes surrounding amniotic fIuid in isotonic isovolumetric condition. In contrast, there is a net efflux of amniotic fluid by osmotic bulk flow, resulting in elevation of $Na^+$ concentration in hypotonic isovolumetric condition. 2) In hypervolumetric conditions, there is a massive efflux of amniotic fluid or solvent drag through the surrounding membranes by fiItrative bulk flow, where the rate of $Na^+$ efflux has a linear relationship with that of water efflux. This is assumed to be carried out through enlarged and newly opened intercellular spaces resulting from increased intraamniotic pressure. 3) Once increasing intraamniotic pressure reaches a point allowing $Li^+$ to pass through during osmotic bulk flow in hypotonic amniotic fIuid, $Na^+$ influx seems to occur by diffusion simultaneously or immediately thereafter, too.
Efficient oocyte activation is a key step for the success of nuclear transfer in cloning. Ionomycin sequentially combined with 6-DMAP is now widely used to activate normal oocytes for analytical studies of oocyte activation and to activate reconstructed oocytes after nuclear transfer. The present study investigated sources of oocytes, duration of ionomycin and 6-DMAP, laser and electric stimulation in goat oocyte activation in order to optimize the protocols. Goat ovaries were collected in individual abattoirs during the breeding season and were delivered to the laboratory within 6 h in saline with 100 IU/ml streptomycin and 0.05 mg/ml penicillin. The oocytes were denuded from the cumulus cell by pipetting with 0.2% hyaluronidase in PBS at 20~22 hr post maturation. Oocytes with the polar body were selected and assigned to four groups for parthenogenetic activation. To examine the effect of duration of ionomycin treatment, oocytes after 20~22 hr of maturation were treated with 2.5 uM ionomycin for 1 or 5 min times and then cultured in 2 mM 6-DMAP for 2 or 4 hr. The activated oocytes were cultured in mSOF at $38.5^{\circ}C$ in $CO_2$ 5%, $O_2$ 5% and $N_2$ 90% multi incubator. Cleavage and blastocyst development was observed at 48 hr and day 8 of culture $in$$vitro$, respectively. Activation rates of oocytes exposed to ionomycin for 1 min(86.4%) were significantly higher than those treated for 5 min(74.3%) duration. This indicated that 1 min ionomycin treatment was most suitable for activation of goat oocytes. The duration of 6-DMAP treat duration was in 2 mM 6-DMAP for 2 hr after 1 min exposure to 2.5 uM ionomycin. The activation rate of oocytes incubated in 6-DMAP for 2 hour(82.5%) was significantly higher than those in oocytes treated with 4 hr(75.5%).
Objectives: This study was performed to analyze the toxicity and to find the lethal dose of the test substance Water-soluble Carthami-flos pharmacopuncture (WCF) when used as a single intravenous-dose in 6-week-old, male and female Sprague-Dawley rats. Methods: The experiment was conducted at Biotoxtech according to Good Laboratory Practices. 20 female and 20 male Spague-Dawley rats were divided into 4 groups of 5 female and 5 male animals per group. The rats in the three experimental groups received single intravenous injections with 0.125-mL, 0.25-mL and 0.5-mL/animal doses of WCF, Groups 2, 3, and 4, respectively, and the control group, Group 1, received a single intravenous injection with a 0.5-mL dose of normal saline. Clinical signs were observed and body weight measurements were carried out for 14 days following the injections. At the end of the observation period, hematology, clinical chemistry, histopathological tests and necropsy were performed on the injected parts. Results: No deaths occurred in any of the groups. Also, no significant changes in body weight, hematological parameters or clinical chemistry test results between the control group and the experimental groups were observed. Visual inspection after necropsy showed no abnormalities. Histopathological tests on the injected parts showed no significant differences, except for Group 1 females; however, the result was spontaneous generation and had no toxicological meaning because it was not dose-dependent. Therefore, this study showed that WCF had no effect on the injected parts in terms of clinical signs, body weight, hematology, clinical chemistry, and necropsy. Conclusion: As a result of single intravenous-dose tests of the test substance WCF in 4 groups of rats, the lethal dose for both males and females exceeded 0.5 mL/animal. Therefore, WCF is a relatively safe pharmacopuncture that can be used for treatment, but further studies should be performed.
Kim, Yu-Jong;Jo, Su-Jeong;Choi, Young-Doo;Kim, Eun-Jung;Kim, Kap-Sung;Lee, Seung-Deok
Journal of Pharmacopuncture
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v.17
no.3
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pp.25-30
/
2014
Objectives: This study was performed to evaluate the single-dose intravenous toxicity of Guseonwangdo-go glucose 20% pharmacopuncture. Methods: Forty Sprague-Dawley rats were divided into four groups of five males and five females per group: an intravenous (IV) injection of 1.0 mL of normal saline solution per animal was administered to group 1 (G1, control group); an IV injections of 0.1, 0.5, and 1.0 mL of Guseonwangdo-go glucose pharmacopuncture per animal were administered to experimental groups 2, 3, and 4 (G2, G3, and G4), respectively. General symptoms, body weights, hematological and biochemical test results, and necropsy histopathological observation were recorded in all groups. In the statistical analyses, significance was determined by using the one-way analysis of variance (ANOVA). The significance level was 0.05 in all comparisons. Results: For 14 days, no deaths or abnormalities were observed in any of the 4 groups. The body weights of all groups continuously increased during the observation period. In the hematological test, the WBC count was significantly increased in female rats of G4 compared to the control group, but this difference was considered not to be statistically meaningful. No significant biochemical changes were observed. On necropsy, crust formation was observed in one rat of the control group, and granulation tissues were observed around the injection site in one rat of G4; these changes were concluded to have been caused by injection of the needle into a vein. Conclusion: The findings suggest that the lethal dose of Guseonwangdo-go glucose pharmacopuncture is more than 1.0 mL per animal in both male and female rats. Thus, we can conclude that Guseonwangdo-go glucose pharmacopuncture injection is relatively safe to use in acute toxicity tests. Further studies are needed to establish more detailed evidences of its toxicity.
Purpose: This study was performed to introduce an in vivo hybrid multimodality technique involving the coregistration of micro-computed tomography (micro-CT) and high-resolution magnetic resonance imaging (HR-MRI) to concomitantly visualize and quantify mineralization and vascularization at follow-up in a rat model. Materials and Methods: Three adult female rats were randomly assigned as test subjects, with 1 rat serving as a control subject. For 20 weeks, the test rats received a weekly intravenous injection of 30 ㎍/kg zoledronic acid, and the control rat was administered a similar dose of normal saline. Bilateral extraction of the lower first and second molars was performed after 10 weeks. All rats were scanned once every 4 weeks with both micro-CT and HR-MRI. Micro-CT and HR-MRI images were registered and fused in the same 3-dimensional region to quantify blood flow velocity and trabecular bone thickness at T0 (baseline), T4 (4 weeks), T8 (8 weeks), T12 (12 weeks), T16 (16 weeks), and T20 (20 weeks). Histological assessment was the gold standard with which the findings were compared. Results: The histomorphometric images at T20 aligned with the HR-MRI findings, with both test and control rats demonstrating reduced trabecular bone vasculature and blood vessel density. The micro-CT findings were also consistent with the histomorphometric changes, which revealed that the test rats had thicker trabecular bone and smaller marrow spaces than the control rat. Conclusion: The combination of micro-CT and HR-MRI may be considered a powerful non-invasive novel technique for the longitudinal quantification of localized mineralization and vascularization.
Objectives : This study was designed to examine the effects of Polygonatum odoratum EtOH ext. on lowering lipid and anti-oxidation using hyperlipidemic rat. Methods : Male rats weighting $195.21{\pm}4.93g$ were divided into 4 groups and fed high fat diet for 8 weeks. Each of 7 rats was divided into a control and sample group. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Polygonatum odoratum(100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of anti-oxidative activity and tumor necrosis factor-$\alpha$($TNF-{\alpha}$). Results : 1. Concentration of plasma free fatty acid, low density lipoprotein-cholesterol showed a significant decrement in the 200 mg/kg and 300 mg/kg Polygonatum odoratum EtOH ext. groups than that of control group. Concentration of plasma total cholesterol, triglyceride showed a significant decrement in all Polygonatum odoratum EtOH ext. groups than that of control group. However, concentration of plasma high density lipoprotein-cholesterol was not significantly different in all the treatment groups. 2. Concentration of liver total cholesterol showed a significant decrement in the 200 mg/kg and 300 mg/kg Polygonatum odoratum EtOH ext. groups than that of control group. Concentration of liver triglyceride(TG) showed a significant decrement in all Polygonatum odoratum EtOH ext. groups than that of control group. 3. Concentration of plasma thiobarbituric acid reactive substance, and liver thiobarbituric acid reactive substance showed a significant decrement in the 200 mg/kg and 300 mg/kg Polygonatum odoratum EtOH ext. groups than that of control group. 4. The values of glutathione peroxidase activity showed a significant increment in all Polygonatum odoratum EtOH ext. groups than that of control group. The values of superoxide dismutase activity and catalase activity showed a significant increment in the 200 mg/kg and 300 mg/kg Polygonatum odoratum EtOH ext. groups than that of control group. 5. The values of plasma aspartate aminotransferase and alanine aminotransferase activities were not significantly different in all treatment groups. 6. Concentration of liver $TNF-{\alpha}$ showed a significant decrement in the 200 mg/kg and 300 mg/kg Polygonatum odoratum EtOH ext. groups than that of control group. Conclusions : Based on the results in this study, the Polygonatum odoratum EtOH ext. showed a positive effect in lowering lipid and anti-oxidation.
Objective : Frutus gardeniae, seed of Gardenia jasminoides Ellis is one of the crude drugs used for the treatment of inflammatory condition in oriental medicine. Methodes : The present study aimed to examine the analgesic effect and anti-inflammatory effect of Frutus gardeniae extract (FGE) on a rat model of ankle sprain pain, and the relations between FGE-induced effect and endogenous nitric oxide (NO) and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord. As a chronic pain model, ankle sprain pain model was used to test the effect of FCE injection applied to acupuncture point. After the induction of ankle sprain, rats subsequently showed a reduced stepping force of the affected limb for at least the next 4 days. The reduced stepping force of the limb was presumably due to a painful knee. FGE dissolved in normal saline was injected several acupoints. Results : After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 8 hours. FGE produced significant improvement of stepping force of the hindlimb affected by the ankle sprain lasting at least 4 hours. FGE produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. In addition, FGE injection showed inhibitory effect on the paw edema induced by ankle sprain. Both NO production and iNOS, COX-2 protein expression increased by ankle sprain were suppressed by FGE. FGE on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the ankle sprain than either FGE or EA did. The present study suggest that FGE produces a potent analgesic effect on the ankle sprain pain model of the rat and that FGE-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.
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