• The Journal of Korean Orthopaedic Ultrasound Society
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• v.5 no.1
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• pp.1-8
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• 2012

Purpose: The purpose of this study is to report the preliminarily result of the radiologic disappearance of the calcific material, regardless of the size, type or location, on one-time ultrasonographic (US) assisted needling for calcific tendinitis of the shoulder. Materials and Methods: From March to August 2011, 46 patients (47 shoulders) with symptomatic calcific tendinitis were treated by one-time US assisted needling. Initially, a diagnostic US was performed with patient to determine the locations, numbers and sizes of calcific deposits. After 1% lidocaine local anesthesia, the calcific material was punctured with an 18-gauge needle under US monitoring. If no calcific material was aspirated after 2 or 3 additional attempts, the deposits was performed multiple puncture to achieve decompression. And then all patients were performed subacromial corticosteroid injection. All patients were followed up 4 weeks after procedure. To assess the radiologic disappearance after one-time US assisted needling, simple radiographs of the treated shoulder were performed and size, dense, and morphology of the calcific deposits were compared with those in baseline radiographs. For clinical evaluations, visual analogue scale for pain and function (PVAS and FVAS), and American Shoulder and Elbow Surgeons (ASES) score were assessed. Results: There were 11 male and 35 female patients with the mean age of 53.8 years (28-71). The morphology of the calcific deposits were 31 type A and 16 type B by French Arthroscopic Society classification and mean size was $2.9{\pm}6.7$ mm before the procedure. At 4 weeks after the index procedure, the radiographic unchanged group was included in 10 cases and changed group was 37 cases. No intergroup difference for the clinical results after the procedure was evident, but group FAS classification before the procedure was significantly different (p=0.011). Conclusion: At 4 weeks after one-time US assisted needling for calcific tendinitis of the shoulder, the radiographic size- or dense-changed cases were showed in 79%, regardless of the size, type or location of the calcific material. But the radiographic nearly or complete disappearance were showed in only 21%.

• Journal of Chest Surgery
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• v.37 no.7
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• pp.547-552
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• 2004

Recently, autologous bone marrow cell transplantation (CTx) for angiogenesis and myogenesis in ischemic myocardium has been extensively investigated to improve heart functions. This study was designed to evaluate the effects of CTx with off-pump coronary artery bypass grafting (OPCAB) in patients who were not feasible for complete revascularization. Material and Method: Four male patients underwent CTx and OPCAB simultaneously. Bone marrow was aspirated from iliac bone. Mean 1.5 ${\times}$ 10$^{9}$ mononuclear cells including mean 6.7 ${\times}$ 10$^{6}$ CD34 + cells and 3.7 ${\times}$ 10$^{6}$ AC133 + cells were obtained and concentrated with 10 cc. These cells were transplanted into non-graftable ischemic myocardium after OPCAB. The heart function of all patients were evaluated using the MIBI scan, echocardiogram and MRI preoperatively. The effects of CTx was evaluated using MIBI scan and echocardiogram at 1 month postoperatively. Result: An average of 2 grafts were bypassed to left anterior descending artery territory. Other territories were transplanted with isolated mononuclear cell. All patients had uncomplicated postoperative course. After 1 month follow up, there were improvement in symptom, ejection fraction (from 49% to 55%) on echocardiogram and myocardial perfusion on MIBI scan in all patients. Conclusion: These preliminary data showed improvement of heart function and myocardial perfusion and also showed the feasibility and safety of combined therapy with OPCAB and CTx in ischemic myocardium. However, the effectiveness of CTx alone cannot be readily assessed. Further randomized, controlled studies are required to evaluate the effectiveness of CTx alone.

• Clinical and Experimental Pediatrics
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• v.49 no.3
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• pp.278-286
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• 2006

Purpose : Although suprapubic bladder aspiration(SBA) is the most reliable technique for identifying bacteriuria in young infant, no report has been published about the clinical manifestations of urinary tract infection(UTI) diagnosed by aspirated urine in Korea. This study was performed to examine clinical manifestations and related factors of UTI confirmed by a positive urine culture obtained by SBA in young infants. Methods : We reviewed medical records of 159 infants younger than six months of age, who had been admitted for UTI to Korea University Ansan Hospital from January 2002 to June 2005. Results : The male:female ratio was 5.1 : 1. The most common pathogen in urine culture was Escherichia coli(92.5 percent). Most of the gram-negative pathogens had high sensitivity to amikacin, or third generation cephalosporins and had low sensitivity to ampicillin, trimethoprim-sulfamethoxazole, or gentamicin. Hydronephrosis and vesicoureteral reflux(VUR) were present in 32.7 percent and 27.7 percent respectively. Renal cortical defects observed on DMSA scans were detected in 37.1 percent. Prolonged fever duration and hospital day, high erythrocyte sedimentation rate(ESR) and C-reactive protein(CRP) levels, hydronephrosis and VUR were related to the renal cortical defects(P<0.05). Rates of UTI-associated bacteremia and aseptic meningitis were 8.3 percent and 6.6 percent. Conclusion : Early differential diagnosis is very important in infants younger than 6 months of age with UTI because the clinical findings are non-specific and UTI-associated bacteremia and aseptic meningitis are concomitantly found. Because prolonged fever and higher ESR and CRP levels are risk factors of the renal cortical defects, radiologic evaluations and nephrologic follow-up were needed in identifying the predisposing congenital abnormalities and chronic renal scarring.

• Childhood Kidney Diseases
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• v.11 no.1
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• pp.59-64
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• 2007

Purpose : Suprapubic aspiration(SPA) has been considered the "gold standard" for obtaining urine in non-toilet trained infants. Ultrasound(US)-guided SPA improves the success rate of the procedure and reduces the complications. However, many physicians perceive SPA as invasive and prefer the use of urethral catheterization (Ucat). We compared the success rate, complications and accuracy of US-guided SPA and Ucat. Methods : 121 infants who visited Ewha Womans University Mokdong Hospital with suspected urinary tract infection(UTI) were investigated. For the first study, the study infants were randomly assigned to either the US-guided SPA(n=32) or Ucat(n=32) groups. The success rate and complications of both procedures were compared. For the second study, US-guided SPA and Ucat were performed simultaneously(n=57). The accuracy of urethral catheterization was subsequently analyzed. The criteria for success was defined as the collection of more than 0.5 mL of urine. UTI was diagnosed by the presence of uropathogens over 105 colony-forming units (CFU)/mL. Results : The overall success rate of the US-guided SPA was 96.9%(71.9% in first attempts, 25.0% in second attempts) which was not significantly different compared to 96.9%(90.6% in first attempts, 6.3% in second attempts) in the Ucat(P>0.05) group. The aspirated urine volume was $7.4{\pm}3.7mL$ in the US-guided SPA group, which was not significantly different to $4.5{\pm}2.6mL$ in the Ucat(P>0.05) group. The accuracy of Ucat in comparison to the US guided SPA was low with sensitivity 59.5%, specificity 86.6%, false-positive rate 13.3% and false-negative rate 40.5%. Conclusion : US-guided SPA should be encouraged as the best method to collect the urine in non-toilet trained infants with UTI.

• Journal of Korean Ophthalmic Optics Society
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• v.7 no.1
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• pp.39-43
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• 2002

This study was investigated the anti-angiogenic activities of Cnidium officinale Makino and Tabanus bovinus by using Chorioallantoic membrane (CAM) and rat cornea. First experiment, the fertilized chicken eggs were kept in a humidified egg incubator at $37^{\circ}C$. After 4 days incubation, about 3 ml of albumin was aspirated from eggs with an hypodermic needles through a small hole drilled at the narrow end of the eggs, and the shell membrane on the floor of the air sac was peeled away. Embryos with chorioallantois of 3~5 mm in diameter were employed for the assay of antiangiogenic activity. Retinoic acid was used as a positive control for this experiment. After 48 h of treatment. branching pattern of blood vessels below coverslip containing retinoic acid ($10{\mu}g/egg$) was dramatically decreased. A simiar angiogenic inhibition was observed in the CAM treated with $50{\mu}g/egg$ of Cnidium omcinale Makino and Tabanus bovinus extracts. Second experiment, rat corneal neovascularization was induced by suturing (one stitch) the cornea with 10-0 nylon, and terramycin was applied on the cornea for I week to prevent corneal inflammation. In the cornea of rats untreated with herbal extracts, numerous vessels were usually seen invading the cornea by day 2 or 3 after suture, and reaching the lesion area within 5~6 days. Corneal neovascularization was gradually increased and peaked at 3 weeks. In contrast to this, herbal extracts conspicuously inhibited the angiogenesis, Oral administration of herbal extracts (20 mg/kg body weight/day) for 4 weeks significantly inhibited the rat corneal angiogenesis induced by suture, and the length of blood vessels in herbal medicine-treated rat cornea was conspicuously lower than that in control animals. A similar phenomenon was also observed in the rat cornea treated with thalidomide (200 mg/kg body weight/day). These findings indicate the anti-angiogenic properties of Cnidium officinale Makino and Tabanus bovinus, suggesting that these properties may be one of the pharmacological mechanisms underlying the anti-tumor and anti-metastatic activities of herbal extracts tested in this study.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.79-87
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• 2005

The ovaries from Hanwoo and Holstein were collected from labattoir and transferred to laboratory. Oocytes were aspirated and incubated in $CO_2$ incubator for 24 hours for maturation. Oocytes were coincubated with the sperms for 5 hours. Cleaved oocytes were selected 48 hours after coincubation and half of the medium was changed newly every 48 hour until blastocyst formation. Cleavage rate and blastocyst rate were investigated according to different breeds and different status of cumulus cells surrounding the oocytes. Blastocysts were either transferred to the recipients or frozen until use. The result of embryo transfer with fresh or frozen embryos was investigated. The rate of male offspring following embryo transfer was also investigated. The rate of cleavage was $66.4\%$ for Hanwoo and $62.4\%$ for Holstein oocytes. The rate of cleavage according to status of oocyte was shown highest in the oocytes completely surrounded with cumulus cells and lowest in denuded oocytes for both Hanwoo and Holstein oocytes. The rate of blastocyst from cleaved oocytes was $40.6\%$ for Hanwoo and $36.9\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with fresh IVF embryos was $57.2\%$ for Hanwoo and $53.3\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with frozen IVF embryos was $40.9\%$ for Hanwoo and $36.4\%$ for Holstein. The rate of male calf produced by embryo transfer was $63.6\%$ for Hanwoo and $50.0\%$ for Holstein.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.163-168
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• 2006

The present study was carried out to investigate in vitro development and post-thawed survivability of bovine embryos according to different ovary transport temperatures. Bovine ovaries were collected at a local slaughterhouse and were transported at 4 different temperature categories to laboratory: $7{\sim}10^{\circ}C\;(T1),\;11{\sim}17^{\circ}C\;(T2),\;18{\sim}25^{\circ}C\;(T3)$ and above $26^{\circ}C$ (control group). The cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured. The rates of maturation (to metaphase II), cleavage and development to blastocysts were compared among treatment groups. Furthermore, frozen-thawed blastocysts were in vitro cultured to compare the survivability among groups. The maturation rates in the T1, T2 and T3 groups ($60.0{\sim}68.2%$) were significantly lower than that in the control group (81.8%, p<0.05). The cleavage rates in the T1 and T2 groups (52.6 and 54.5%) were significantly lower than that in the control group (83.6%, p<0.05). However, there was no difference in the development rate to blastocysts among all groups ($27.9{\sim}33.0%$, p>0.05). The survivability of frozen-thawed embryos was significantly lower in the T1 group (46.2%) than those in the T2, T3 and control groups ($68.8{\sim}7.13%$, p<0.05). In conclusion, the results suggest that ovary transport temperature at $26^{\circ}C$ may be optimal for the better in vitro development and the survival of frozen-thawed embryos produced in vitro Furthermore, exposure of ovary to temperature below $10^{\circ}C$ during transport may significantly decrease both in vitro development and survivability of frozen-thawed blastocysts.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.255-262
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• 2006

The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.9-20
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• 1999

The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.