• Journal of Embryo Transfer
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• v.4 no.1
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• pp.28-34
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• 1989

These experiments were carried out to determine the effect of cell stage in embryo bisection on the sub-Sequent in vitro and in vivo development in mouse. The embryos of ICR mouse were microsurgicaily bisected at 2-cell, 4-cell, 8-cell, morula and blastocyst stage using a microsurgical blade attached a micromanipulator. These demi-embryos without zona pellucida were cultured up to blastocyst stage and transferred to pseudopregnant mice, and the development of these demi-embryos was compared with the results of intact embryos of the corresponding cell stage. The successful rate of mouse embryo bisection at 4-cell stage (59.0%) was significantly (p <0.05) lower than those at 8-cell (75.6%), 2ce11 (80.7%) or morula stage (84.8%), and highest at blastocyst stage (95.7%). When the bisected embryos without any damage from microsurgery were cultured in vitro up to blastocyst,the in vitro de'velopment of demi-embroys bisected at morula to blastocyst was 91.6 to 95.3%, which was similar to the culture result of intact embryos of corresponding stage. However, the in vitro development of demi-em-bryos bisected at 2- to 8-cell stage was signiflcantiy (p <0.05) lower.The post-transfer implantation rate of demi-embryos developed in vitro to eu-blastocyst were 19.6 and 25.4% in demi-embryos bisected at morula and blastocyst stage,respectively and not significantly (P <0.05)different from the result of intact embryos of the same stage. However, the implantation rates of demi-embryos bisected at 2- or 8-cell stage were significantly (P <0.05) lower than the result from the intact embryos of the corresponding stage.

• Journal of Embryo Transfer
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• v.13 no.3
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• pp.219-226
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• 1998

Enucleation of oocytes is an important limiting step for embryo cloning. We propose an enucleation technique based on the removal of chromatin after oocyte activation by aspirating the second polar body containing complemented chromatin. In a preliminary experiment to determine an optimal age of oocytes enucleation in rabbits, oocytes were enucleated at 15~20 hours post hCG. Recently ovulated oocytes were enucleated at a higher rate than aged oocytes. Microsurgical removal of the complemented chromatin in the second polar body was significantly more effective in enucleating than aspiration of a larger cytoplasm volume surrounding the first polar body of metaphase-arrested oocytes(96.8% versus 70.4%; P〈0.05). Moreover, compared with a nuclear transplantation protocol based on enucleation of metaphase-arrested oocytes and preactivated oocytes followed by treatment with 5 $\mu$M ionomycin for 5 min and 2 mM DMAP for 1 hr, there was no significant difference in the rate of blastocyst development. The ease with which modified technique can be performed is likely to render this technique widely useful for research and practice on mammalian cloning.

• Archives of Plastic Surgery
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• v.38 no.2
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• pp.212-216
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• 2011

Purpose: Ameloblastomas are rare benign tumors of odontogenic origin, and compose about 1% of all oral and maxillomandibular cysts and tumors. Because this neoplasm has a high rate of local recurrence, segmental mandibulectomy with a 1~2 cm safety margin and immediate microsurgical reconstruction is an accepted treatment modality. The authors experienced four mandibular reconstruction cases that underwent secondary dental implantation. Here, the authors describe these cases and their long-term results. Methods: Four patients with ameloblastoma of the mandible underwent segmental mandibulectomy and reconstruction with a free fibula osseous flap from January 1999 to May 2005, followed by secondary dental implantation. Recurrence, bony union, implant osseointegration, and functional and aesthetic results were evaluated by radiologic imaging, by physical examination, and by using photographs. Results: All free flaps survived with no evidence of flap loss. To date, no recurrence has been noted clinically or radiologically. Imaging after mandibular reconstruction with a free fibular flap revealed satisfactory bony unions and mandibular contours. The patients achieved good aesthetic and functional results after the secondary implantation. Conclusion: Mandibular reconstruction using a fibular osseous flap and secondary dental implantation can produce good functional and aesthetic results after segmental mandibulectomy for ameloblastoma.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.229-238
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• 1997

Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$\times$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$\pm$0.74 and 15.2$\pm$1.32${\mu}{\textrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.

• Archives of Reconstructive Microsurgery
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• v.16 no.2
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• pp.100-107
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• 2007

Treatment of giant cell tumor of distal radius can be treated in several ways according to the aggressiveness of the tumor. But the management of giant cell tumor involving juxta-articular portion has always been a difficult problem. In some giant cell tumors with bony destruction, a wide segmental resection may be needed for preventing to recur. But a main problem is preserving of bony continuity in bony defect as well as preservation of joint function. We have attempted to overcome these problems by using a microvascular technique to transfer the fibula with peroneal vascular pedicle or anterior tibial vessel as living bone graft. From April 1984 to July 2005, we performed the reconstruction of wide bone defect after segmental resection of giant cell tumor in 14 cases, using Vascularized Fibular Graft, which occur at the distal radius. VFG with peroneal vascular pedicle was in 8 cases and anterior tibial vessel was 6 cases. Recipient artery was radial artery in all cases. Method of connection was end to end anastomosis in 11 cases, and end to side in 3 cases. An average follow-up was 6 years 6 months, average bone defect after wide segmental resection of lesion was 6.8 cm. All cases revealed good bony union in average 6.5 months, and we got the wide range of motion of wrist joint without recurrence and serious complications. Grafted bone was all alive. In functional analysis, there was good in 7 cases, fair in 4 cases and bad in 1 case. Pain was decreased in all cases but there was nearly normal joint in only 4 cases. Vascularized fibular graft around wrist joint provided good functional restoration without local recurrence.

• Archives of Reconstructive Microsurgery
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• v.13 no.1
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• pp.24-28
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• 2004

The vascularity of a skin island in fibula osteocutaneous free flap often depends on musculocutaneous perforators that find their origin in the proximal peroneal artery. But a potential drawback has been reported on the unreliability of the skin paddle. The perforating vessels to the skin paddle of the fibula osteocutaneous free flap were rarely derived from a common tibio-fibula trunk, an anterior tibial artery and a posterior tibial artery. Previous studies have emphasized total loss of the overlying skin paddle, if the expected perforating vessels are not present. We report here on our experience that the skin paddle of the fibula osteocutaneous free flap was vascularized not by a peroneal artery but a direct branch of the posterior tibial artery. There were no intraseptal nor intramuscular pedicles in the posterior crural septum which connected to the overlying skin island. Therefore, we performed microsurgical anastomoses between distal peroneal vessels of the fibula and the perforating branches of the posterior tibial vessels of the skin paddle. The anastomosed skin paddle was salvaged with a peroneal flow through vascular anastomosis and was transferred to the bone and intraoral soft tissue defects with the fibula graft. The patient had no evidence of vascular compromise in the postoperative period and showed good healing of the intraoral skin flap.

• Archives of Reconstructive Microsurgery
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• v.13 no.1
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• pp.58-62
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• 2004

Purpose: To cover the exposed tendons and bones after trauma and cure the concomitant osteomyelitis in the lower extremities, gracilis muscle free flaps are frequently preferred. 32 cases of gracilis muscle free flap we had done were analysed according to the indications, specification of flap length and width, pedicle length, vessels used in the anastomosis and final healing after at least over 1 year follow up. Materials and methods: From August 1995 through November 2002, we have performed 32 cases of gracilis muscle free flap transplantation with the general microsurgical procedures in the lower extremities. Open fracture of the middle and distal tibia were 12, exposed heel 6, crushing injury in the foot 5 cases, open fracture of the ankle 4, chronic osteomyelitis of the tibia 3 and osteomyelitis of the tarsal bones 2. Tailored flap length were ranged from 16 cm to 4 cm, width were from 5cm to 4cm. Pedicle length averaged around 4 cm. Anastomosis of one artery and two veins in both of donor and recipient were performed in 17 cases and one artery and one vein in 15 cases. Results: All flaps were survived, except 2 cases. Final flap healing was satisfactory to both of the patients and microsurgeon. Conclusion: Gracilis muscle free flaps are frequently chosen to cover the exposed components and cure the osteomyelitis in the lower extremities.

• Archives of Reconstructive Microsurgery
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• v.13 no.1
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• pp.1-6
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• 2004

Advances in microvascular techniques and refinements in microsurgical tissue transfers have enabled surgeons to combine different tissue components and reconstruction into a single-stage operation in extensive or composite defect following injury. Some problems and consideration for extensive or composite defects are form, shape, function, and dimension of the defect sites. Therefore combination of two or more flaps is required to reconstruct extensive or composite defect. This paper presents our clinical experience of four cases of combined free flaps with or without sequential microanastomosis in reconstruction of upper extremity based on peroneal flowthrough, thoracodorsal, and dorsalis pedis vascular system. Satisfactory results were obtained without flap loss and complications. The free flaps were combined in th following fashion; two cases by bridge fashion, one by chimeric microanastomosis and one by simple chimeric fashion. The median follow-up time on all patients was 21.7 months. Donor site morbidity was minimal. Extensive soft tissue or composite defects can be effectively covered by various combined flaps. Even though the risk for complication exists, the options of combination with or without sequential microanastomosis can add a functional or sensory dimension to reconstruction of complex wounds.

• Archives of Reconstructive Microsurgery
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• v.17 no.2
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• pp.101-107
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• 2008

Purpose: Free flaps from the great toe are an established method for reconstruction of absent or partially amputated thumbs. However, options differ as to which technique represents the ideal solution for each level of amputation. Various methods of distal thumb reconstruction have been proposed. We prefer to transplant the entire great toe nail complex with the almost all of the pulp rather than a portion of the nail. This paper reflects our experience in using the great toe mini wrap-around flap for distal thumb reconstruction. Materials and Methods: In the period from October of 2005 to July of 2007, 9 patients were treated for traumatic thumb defects localized at the distal phalanx of the thumb. The patients included seven men and two women. The mean age was 44 years (range, 21~60) and the dominant right hand was involved in seven of nine patients. Results: The transferred flaps have survived completely in all cases. The mean range of motion in the interphalangeal joints was 51o, with 73% of the normal uninjured opposite hands. The two-point discrimination was 10.5 mm (range, 5~13 mm). In Semmes-Weinstein monofilament test, the sensibility was 4.31 in 4 cases, 3.61 in 3 cases and 2.83 in 2 cases. The pinch power was 64% (range, 55~95%) of the opposite hand. All patients were satisfied with the appearance of the reconstructed thumb and felt comfortable at final follow-up. Conclusion: We have successfully reconstructed 9 cases of traumatic distal thumb defects using the mini wrap-around free flap. The mini wrap-around free flap in great toe is an excellent alternative method for distal thumb reconstruction in selected patients.

• Archives of Reconstructive Microsurgery
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• v.17 no.2
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• pp.82-86
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• 2008

Purpose: Soft tissue defect can occur on the posterior aspect of the elbow after trauma or fracture fixation. To cover the defect and maintain elbow functions, various flap surgeries including latissimus dorsi muscle flap, lateral arm flap and radial forearm flap can be performed. We present the clinical results of transposition lateral arm flap for coverage of the elbow defect and discuss the cause of posterior soft tissue necrosis after fracture fixation. Materials and Methods: Two patients who had posterior soft tissue defect of the elbow after open reduction of the fractures around the elbow were treated with transposition lateral arm flap. The mean size of skin defect was 20 $cm^2$. The flap was elevated with posterior radial collateral artery pedicle and transposed to the defect area. Donor defect was covered with split thickness skin graft. The elbow was immobilized for 1 week in extended position and active range of motion was permitted. Results: All two cases of transposition lateral arm flap survived without marginal necrosis. The average range of motion of the elbow was 10~115 degrees. Mayo elbow performance score was 72 and Korean DASH score was 23. Conclusion: When elbow fractures are fixed with three simultaneous plates and screws, skin necrosis can occur on the posterior aspect of the elbow around olecranon area. If the size of skin defect is relatively small, transposition lateral arm flap is very useful option for orthopaedic surgeons without microsurgical technique.