• Journal of Embryo Transfer
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• v.13 no.2
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• pp.107-115
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• 1998

The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.0$\pm$11.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.7$\pm$1.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1210-1217
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• 1999

This study was carried out to develop a newly designed ovum pick-up(OPU) instrument for ultrasound-guided transvaginal follicular aspiration in cows. This new instrument consists of out- & inner-layer stainless pipes and a grip with a trigger(hand) switch. Some gauge types of disposable needles and tubes can be attached to this inner pipe. With this instrument, while grasping an ovary with one hand, the other hand can handle in apiration and vacuum on/off with the least assitant's help. With this instrument the mean recovery rate of bovine follicular oocytes was 45.2%. In recovered oocytes, usable oocytes(Grade I & II) were 30.4% and this rate meant 1.4 oocytes per ovary. For 30 days after initial aspiration with this instrument, some adverse effects such as adhesion, hemorrhage, hematoma and other mass formation in/with ovaries were also examined by rectal examination, ultrasonographic and endoscopic images. Adhesion was found in one ovary 1 week after aspiration, and hemorrhagic lesion was found 1-2 days and petechia were found 3-5 days after aspiration and there was no remarkable adverse effects. It was found that this instrument could be applicable and safe for ovum pick-up in cows.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.2
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• pp.115-121
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• 1990

Ultrasonically guided oocyte collection gradually replaces laparoscope in many IVF center. In present study, we compare the efficacy of both methods in our IVF program. Totally 377 cycles which were undertaken in vitro fertilization treatment were divided into 2 groups. Ultrasonically guided transvaginal follicular aspiration was performed in 188 cycles and laparoscopic follicular aspiration was performed in 189 cycles under local anesthesisa. The mean age for both groups was similar. Follicular recruitment was achieved with human menopausal gonadotropin (hMG) or a com bination of clomiphene citrate and hMG or a combination of FSH and hMG. In the ultrasonically guided aspiration group, 1821 follicles were aspirated with 61.8% of recovery rate (1125 oocytes), 81.5% of embryo transfer rate (145 cycles) and (17%), 26 cases intrauterine pregnancies were estabilished. In the laparoscopic group, 604 follicles were aspirated with 68.7% recovery rate (445 oocytes) and a 79.9% ET rate (127 cycles), 11 cases (8.7%) intrauterine pregnancies were estabilished. A valid comparison of these data is not possible because the 2 groups are dissimilar for factors known to influence oocyte development and recovery. No statistically significant differences could be demonstrated between 2 groups in all but the recovery rate and clinical pregnancy rate, In ultrasound group, the clinical pregnancy rate was significantly higher than that of laparoscope group. The potentially detrimental effect of CO2 pnemoperitonium present during laparoscope but not in ultrasound guided recovery on ova quality may underlie the observed difference in the clinical pregnancy rate between the 2 groups. Ultrasound guided aspiration seems to be as effective as laparoscopy in terms of oocyte retrieval and conception rate. Furthermore, the procedure is simple and inexpensive, it may replace laparoscopy as a method for oocyte collection in most patients who undergo IVF.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.147-156
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• 1997

Ultrasound-guided follicular aspiration was performed in Holstein heifers once weekly with or without pretreatment of single or multiple decreasing doses using a total of 400 mg FSH. Oocytes were aspirated with a 6.5 MHz convex-array ultrasound trasducer designed for intravaginal use. All the visible follicles larger than 4 mm in diameter were punctured with a 17 gauge, 55 cm needle at each aspiration session and the follicular fluids containing oocytes were obtained by vacuum suction. The results obtained were as follows: As a preliminary experiment, the recovery rates of folicular oocytes by ultrasound-guided aspiration from the isolated ovaries of Korean native cows were compared between suction methods using manual syringe or vacuum pump. The recovery rate of oocytes using vacuum pump (80.7%) was significantly (P<0.05) higher than that using manual syringe (47.1%). The follicles were counted by their size in diameter with ultrasound image, and recovery rates and grades of follicular oocytes collected by ultrasound-guided aspiration were investigated in Holstein heifers pretreated with or without FSH. A group of heifiers were injected with multiple decreasing doses (twice a day for 3 days) of a total of 400 mg FSH. The other 2 groups were injected with a single dose of 400 mg FSH mixed with 25% PVP. Ultrasound observation of follicle population and/or ultrasound-guided transvaginal oocyte aspiration were performed 12 hrs following the last FSH injection in the multiple dose group, and 48 or 60 hrs after FSH injection in the single dose groups. Most of the visible follicles had small size of less than 3 mm in diameter in unstimulated heifers (71.0%), but medium size in all the heifers treated with FSH. (70.5 to 92.8%). The number of OPU follicles per session (4.6$\pm$1.9) were much less, compared to the vilsible follicle counts (9.7$\pm$2.2), in the nustimulated heifers due to the small dominant follicles. Among 4 goups of heifers the most visible as well as OPU follicles were observed in the heifers at 60 hrs following treatment of a single dose of 400 mg FSH (21.2$\pm$2.3 and 21.0$\pm$2.0), and the differences in both the follicle counts between the groups was found significant (P<0.05) The rates of oocyte recovery from the follicles by ultrasound-guilded aspiration were varied 46.3 to 75.0% in the heifers unstimulated and treated with a single dose of 400 mg FSH, but the group difference was not significant. The number of recovered oocytes per session a, pp.ared to be highest at aspiration at 60 hrs following single FSH (10.6$\pm$2.2) than at aspiration at 48 hrs after single FSH (7.8$\pm$2.7) or in the unstimulated heifers (3.4$\pm$3.0). The proportion of grade I and II oocytes to all oocytes collected was varied 31.8 to 64.0% between the groups. However, there was found no significant difference in both the number of oocytes recovered per session and the percentage and the percentage of grade I and II oocytes. From the above results it was concluded that the more oocytes of superior quality might be recovered economically by ultrasound-guided aspiration at 60 hrs following the pretreatment of a single dose of 400 mg FSH and by suction using a vacuum pump system of about negative pressure of 75 to 85 mmHg.

• Journal of Embryo Transfer
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• v.14 no.3
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• pp.203-210
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• 1999

In this stuyd, the effect of the dominant follicle aspiration for the superovulatory response in HanWoo was investigated. The criterion for the presence or absence of a dominant follicle based on their morphological examination. The dominant follicle was aspirated 48hr before the onset of superovulation treatment by 6.5MHz convex probe connected with a carrier and superovulation induced by FSH (Super-Ov Tyrer, Texas, U.S.A) adminstered twic a day s.c. over 4 day in a decreasing regimen. From 13 HanWoo scanned daily to determine the presence and growth of the dominant follicle, its an average diameter of 15.4mm was measured and an average diameter of corpora lutea was 18.7mm on day of follicular aspiration. In the experiment, a follicular remove by ultrasound-guided aspiration, the ovarian response was significantly enhanced when animals were superovulated in the aspiation of a dominant follicle compare with animals superovulated non-aspiration of a dominat follicle. In the aspiration of a dominant follicle donors yieleded more corpora lutea(14.4$\pm$4.7 vs 8.6$\pm$3.4) and transferable embryos(8.9$\pm$4.2 vs 5.4$\pm$2.7) than control. In cows in which the dominant follicle had been aspirated under sonographical control 2 days before superovuation, the number of corpus lutea and transferable embryos were significantly enhanced compared with animals superovulated in the presence of a dominant follicle (14.4$\pm$4.7 vs 6.9$\pm$2.7, ; 8.9$\pm$4.2 vs 3.3$\pm$1.6). After 7 days of artificial insemination, the embryos at 7 days were cllected by uterine flushing after dominant follicle insemination, the embryos at 7 days were collected by uterine flushing after dominant follicle aspiration and superovulation treatment, and evaluated their quality by morphological criteria. Sixteen embryos with excellent and good grade were transferred into 8 recipient cows. Six pregnancies were identified at 60 and 120 days of gestation by rectal palpations. In conclusion, the present study showed that 1) the presence or absence of a dominant follicle signficicnatly affects superovulatory responses, and 2) ultrasound-guided follicular aspiration of the dominant follicle and superovuation treatment provides an accurate and procedure to increase ovarian responses in HanWoo.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.121-129
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• 1999

The objective of this study was to produce calves by transfer of embryos derived from slaughter house(SHD) and ultrasound-guided ovum pick-up (OPU). At 60 hrs after injection of 400 mg FSH dissolved in 25% polyvinylpyrrolidone(PVP) by single dose, ultrasound-guided follicular oocyte aspiration was ferformed. Day-7 and day-8 blastocysts produced by in vitro maturation (IVM), fertilization (IVF) and culture(IVC) of the oocytes derived from SHD and OPU were nonsurgically transferred into recipients. The results obtained were as follows. The cleavage rate and the development rate to blastocysts were not significantly (P<0.05) different between the oocytes obtained by SHD (72.9% vs. 34.1%) and OPU (75.9% vs. 38.4%). The oocyte recovery rate from the number of follicles by ultrasound-guided aspiration were not significantly (P<0.05) different between Holstein (61.7%) and Hanwoo (60.1%), but the rate of oocytes useful for IVF was significantly (P<0.05) higher in Hanwoo (69.3%) than Holstein (59.6%). The cleavage rate and the development rate to blastocysts was not significantly (P<0.05) different between Holstein (74.9% vs. 39.2%) and recipients on day 8 of estrus cycle resulted in 13 pregnancies (34.2%). One of them was sacrificed during gestation period due to mastitis and another was aborted spontaneous. The resulting 14 calves were morphologically normal at birth. Seventy eight fresh OPU-IVF embryos were transferred into 21 recipients on day 8 of estrus cycles, resulting in pregnancy of 12 recipients (41.4%). Two of them were sacrificed during gestation period due to mastitis and the other two were aborted. Nevertheless, the 11 OPU-calves have been born normally.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.195-202
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• 1997

The present study was carried out to evaluate the effect of superovulation treatments on ovarian responses, oocyte recovery rates and grades of collected oocytes using an ultrasound-guided transvaginal approach in Korean native cows. Superovulation in cows was induced with two different regimenes: 1) FSH-decreasing dose(n=8): the cows were received twice per day for three days of the total dose of 400 mg of FSH-p, 2) FSH-single dose(n=9): the cows were administrated a single dose of 400 mg of FSH-p in 25% PVP. The Observation of visible follicles and collection of oocytes were performed 12 hours following the last FSH in FSH-decreasing dose group and 48 hours after the FSH-single dose injection. All visible follicles larger than 6 mm were punctured and aspirated with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use. The mean number of visible follicles(> 6 mm) was significantly(P<0.05) higher in the FSH-decreasing dose treatment (22.811.9) and FSH-single dose treatment (20.612.0) groups than the non-treatment group(7.0$\pm$8). The mean recovery rate of oocytes was not significantly(P<0.05) different between the treatment and control groups, but the mean number of collected oocytes was significantly(P<0.05) higher in the FSH-decreasing dose treatment( 12.611.5) and FSH-single dose treatment (11.813.6) groups than the non-treatment group(3.7$\pm$0.5). In conclusion, the FSH-single dose treatment at superovulation in cows for ultrasound-guided aspiration might increase the number of aspiratable follicles and the recovery rate of follicular oocytes as the FSH-decreasing dose treatment.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.199-208
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• 2000

This study was carried out to investigate of factors concerning to ultrasound-guided follicular aspiration; level of vacuum pressure, diameter of use needle, effect of FSH hormone and conception rate after embryos transfer. 1. Oocytes collection number were 4.2$\pm$2.9 e.a to luteuml phase and follicular phase were 4.4$\pm$3.5 e.a to ovaries of Hanwoo. 2. We taked proper level of aspiration vacuum pressure was 40~120 mmHg to oocytes collection. Oocytes collected number were 4.2$\pm$3.2, 4.3$\pm$3.4, 4.5$\pm$3.4 e.a. to 40, 80, 120mmHg, respectively, follicles aspiration rate were 49, 47, 45%. 3. Effect of collection needle diameter was not difference significantly(P＜0.05), oocytes collected number were 4.4$\pm$3.5 e.a to 170 and 3.0$\pm$1.8 e.a to 18G needle, collected oocytes quality were no difference significantly (P＜0.05). 4. Follicles increase number to FSH hormone injection were 6.2$\pm$2.3 e.a to intramuscle and 1.1$\pm$2.7 e.a. to epithelial injection method. 5. Conception rate derived from E.T. was 11.1% to freezing embryos and 46.2% to fresh E.T., difference significantly(P＜0.05).

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4169-4173
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• 2014

Background: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is highly accurate in diagnosing mediastinal lymphadenopathies of lung cancer and benign disorders. However, the utility of EBUS-TBNA in the diagnosis of mediastinal lymphomas is unclear. The aim of this study was to determine the diagnostic value of EBUS-TBNA in patients with suspected lymphoma. Materials and Methods: Sixty-eight patients with isolated mediastinal lymphadenopathy and suspected of lymphoma were included in the study. EBUS-TBNA was performed on outpatients under moderate sedation. The sensitivity, specificity, negative predictive value and diagnostic accuracy of EBUS-TBNA were calculated. Results: Sixty-four patients were diagnosed by EBUS-TBNA, but four patients with non-diagnostic EBUS-TBNA required surgical procedures. Thirty-five (51.5%) patients had sarcoidosis, six (8.8%) had reactive lymphadenopathy, nine (13.3%) had tuberculosis, one (1.5%) had squamous cell carcinoma, two (2.9%) had sarcoma and fifteen (22%) had lymphoma (follicular center cell, large B-cell primary, and Hodgkin lymphomas in three, two, and ten, respectively). Of the 15 lymphoma patients, thirteen were diagnosed by EBUS and two by thoracotomy and mediastinoscopy. The sensitivity, specificity, negative predictive value, and diagnostic accuracy of EBUS-TBNA for the diagnosis of lymphoma were calculated as 86.7%, 100%, 96.4%, and 97%, respectively. Conclusions: EBUS-TBNA can be employed in the diagnosis of mediastinal lymphoma, instead of more invasive surgical procedures.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.211-218
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• 1997

This experiment was carried out to develop the anesthetic methods for ultrasonography and a new simplified disposable needle guidance device for ovum pick-up(OPU) in cows. Three different anesthetic methods were applied as. 1) epidural analgesia only with 2% lidocaine(20~30 ml), 2) epidural analgesia with 2% under general sedation with xylazine, 3) epidural analgesia with 2% lidocaine under general sedation with detomidine. We evaluated the anesthetic effects with items such as relaxation of anal sphincter, tail movement and rectal wall, retractability of both ovaries, additional anesthesia and possibility of OPU. Through this experiment, the above three anesthetic methods were applicable to OPU, but the epidural anlagesia under general sedation with detomldine was most effective for OPU. We developed a new disposable needle guidance device with stainless steel tube. With this, disposable needles can be easily attatchable to any other intravaginal probes. And also, it was found to he practical, economic and effective for OPU with the recovery rate of 51.2%.