No abstract available.
Fertilization in Vitro* ; Humans* ; Micromanipulation* ; Pregnancy Rate* ; Pregnancy*

OBJECTIVEBoth microsurgical subinguinal varicocelectomy (MSIV) and microsurgical high inguinal varicocelectomy (MHIV) are recommended for the treatment of varicocele, but they differ in technical complexity. This study aimed to determine the microanatomy of spermatic blood vessels in the two surgical approaches.

METHODSWe recorded the numbers of spermatic veins, arteries and lymphatics in 80 cases of MSIV and 20 cases of MHIV. We also examined the spermatic cords from 10 adult male cadavers by histological staining.

RESULTSThe numbers of medium spermatic veins (2 -5 mm in diameter) were 1.80 +/- 0.83 and 3.98 +/- 1. 99 in MHIV and MSIV, respectively, with significant difference between the two groups (t = -7.536, P < 0.01), and the total numbers of spermatic veins were 6.40 +/- 1.67 and 9.01 +/- 2.70, also with significant difference between the two (t = -4.071, P < 0.01). However, there were no significant differences between MHIV and MSIV in the numbers of small spermatic veins (diameter < or = 2 mm), large spermatic veins (diameter > or = 5 mm), arteries and lymphatics, nor in the numbers of spermatic veins and arteries of the cadavers.

CONCLUSIONThe total number of spermatic veins and the number of medium spermatic veins may be larger in MSIV than in MHIV, but the medium spermatic veins do not increase surgical difficulty, and MSIV is not more complicated than MHIV.

Adult ; Arteries ; anatomy & histology ; Humans ; Male ; Micromanipulation ; Microsurgery ; Middle Aged ; Spermatic Cord ; anatomy & histology ; blood supply ; Varicocele ; pathology ; surgery ; Veins ; anatomy & histology ; Young Adult

Although the fertilization rate exceeds to 80~90% with much progress in vitro fertilizaton and embyo transfer(IVF-ET) program, the prgnancy rate rmains at 20~30%, and the endometrial implantaion rate per embryo transferred at 10~15%. Recently, many attempts have been made to improve embrynic implantion after IVF-ET including serveral procedures of assisted hatching(AH) using micromanipulation, and pregnacies and births have been obtained after AH. This clinical study was performed to develop and estabilish AH as an effective procedure to improve embryonic implantioan in IVF-ET patients who had previous repeated failure of standard IVF-ET more than 2 times(Group R), were more than 37 years old(Group A), or had high basal serun FSH levels more than 15 mIU/ml(Group F). From January, 1995 to Februry, 1996, 132 cycles of AH using partial zona dissection(PZD) were performed in 104 infertile patients, and the outcomes of AH were analyzed according to pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation(COH) was 9.9+/-7.1 in 71 cycles of 54 patients who had previous repeated failure more than two times(Group I: Group R,R+A,R+F, and R+A+F), 8.4+/-5.9 in 62 cycles of 46 patients whose age was more than 37 years old(Group II : Groups A, R+A, A+F, and R+A+F), and 8.7+/-6.5 in 49 cycles of 47 patients who had high basal serum FSH levels more than 15 mIU/ml(Group III:Groups F,R+F, A+F, and R+A+F). The number of embroys transferred after AH was 4.7 +/-1.8 in Group I, 4.2 +/-1.9 in Group II, and 4.2+/-2.0 in Froup III. The mean cumulative embryo score(CES) was 56.8+/-30.0 in Group I, 52.6+/-30.6 in Group II, and 52.6+/-29.9 in Group III. There were no significant differences in the numbers of oocytes rerieved and embryos transferred, and CES among 3 groups. The overall clinical pregnancy rate was 14.4%(19/132) per cycle and 18.3%(19/104) per patient. THe clinical pregnancy rate per cycle and per patient was 12.7%(9/71) and 16.7%(9/54) in Group I, 4.8% (3/62) and 6.5%(3/46) in Group II, and 26.5%(13/49) and 27.7%(13/47) in Group III, and there was a significant difference between Group II and Group III. In conclusion, AH of human embryos using micromanipulation might be promising for IVF-ET patients, especially with the past history of repeated failure, old age, and high basal serum FSH level and AH will provide a range of novel techiques which may dramatically improve the implanatation and pregnancy rates in IVF-ET program and contribute much to effective management of infertile couples.
Embryo Transfer* ; Embryonic Structures* ; Family Characteristics ; Fertilization ; Herpes Zoster ; Humans ; Micromanipulation* ; Oocytes ; Parturition ; Pregnancy Rate

In spite of much progress in in vitro fertilization and embryo transfer(IVF-ET) program,the pregnancy rate remains at 20~30%, and the endometrial implantation rate per embryotransferred at 10%. Although IVF-ET is widely applied in the treatment of coupleswith male factor infertility, it may fail in many infertile couples with normal semen parameters,and certain couples cannot be accepted for standard IVF-ET due to unfertilization orextremely low fertilization rate of oocytes. Recently, several procedures of microassistedfertilization(MAF) using micromanipulation have been introduced, and pregnancies and birthshave been obtained after intracytoplasmic sperm injection(ICSI).This clinical study was performed to develop and establish ICSI as an effective procedureof MAF in infertile couples who could not be accepted for standard IVF-ET becauseof extremely impaired semen characteristics(Group A) and because of failure in fertilizationof extremely low fertilization rate of oocytes with the conventional fertilization technique inthe previous IVF-ET cycles(Group B). From March, 1995 to December, 1996, a total of 114cycles of IVF-ET with ICSI in 65 infertile patients were included in study group, and theoutcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score(CES), and pregnancy rate.In Group A, 34 patients were evaluated with semen score such as number of totalmotile sperms, and then divided into 4 groups accordingly. In 62 ICSI cycles, the numberof oocytes retrieved after controlled ovarian hyperstimulation(COH) was 12.4+/-6.8, and thenumber of oocytes optimal for ICSI procedure was 8.8+/-5.5. The fertilization rate of 65.7+/-23.6% could be obtained after ICSI. The number of embryos transferred was 4.4+/-2.2 withthe mean CES of 50.5+/-34.3 in ICSI cycles. The overall pregnancy rate was 24.2%(15/62)per cycle and 44.1%(15/34) per patient. There were no significant differences in the pregnancyrates among 4 groups. Although more mature oocytes were retrieved, the fertilizationrate was significantly lower in Group A-1 compared with Group A-IV. However, semenscore did not clearly affect the outcomes of ICSI in couples with severe male factor infertility.In Group B, the number of oocytes retrieved after COH was 10.5+/-6.1 in 49 previouscycles, and 10.8+/-5.7 in 52 ICSI cycles. In ICSI cycles, the number of oocytes optimal forICSI procedure was 8.5+/-5.1 with the fertilization rate of 72.4+/-22.5%. The number ofembryos transferred was 1.4+/-2.4 in previous cycles, and 4.7+/-1.8 with the mean CES of 50.4+/-29.9 in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 30.8%(16/52) percycles and 51.6%(16/31) per patients.In conclusion, MAF of human oocytes with ICSI is a promising fertilization method forIVF-ET patients, especially with few spermatozoa for the conventional methods of in vitroinsemination and with the past history of failure in fertilization or low fertilization rate inthe previous cycles, and ICSI using micromanipulation procedures applied to human oocyteswill provide a range of novel techniques which may dramatically improve the pregnancy ratein IVF-ET program and contribute much to the effective management of infertile couples.
Embryonic Structures ; Family Characteristics ; Fertilization in Vitro ; Fertilization* ; Humans* ; Infertility ; Male ; Micromanipulation ; Oocytes* ; Pregnancy ; Pregnancy Rate ; Semen ; Sperm Injections, Intracytoplasmic* ; Spermatozoa

This paper deals with the manufacturing state of the art of biochip, and introduces a new method--laser microtechnology, including its developing procedure, characteristics and function in biochip production.
Biosensing Techniques ; instrumentation ; methods ; Computer-Aided Design ; Equipment Design ; Lab-On-A-Chip Devices ; Lasers ; Microchip Analytical Procedures ; methods ; Micromanipulation

OBJECTIVE: We used nucleated erythrocytes in maternal blood for prenatal determination of the fetal gender as the preliminary experiment for the screening of fetal genetic status and the BclI DNA polymorphism in an attempt to clarify the origin of erythrocytes in maternal blood. METHODS: In seventeen pregnant women, venous blood was withdrawn and the nucleated erythrocytes were recovered by magnetic activated cell sorting (MACS) and immunostaining. After isolation of nucleated erythrocytes by micromanipulation, we performed nested PCR for amelogenin gene to identify the fetal gender and performed BclI DNA polymorphism to clarify the origin of erythrocytes. RESULTS: We could amplify the minute DNA in a single cell by primer extension preamplification and nested PCR of amelogenin gene in 94 (48.7%) cells and could identify the fetal gender by 58.8%. BclI DNA polymorphism revealed that the several cells, which did not reveal the specific band of Y chromosome in spite of the pregnancy of male fetuses, must be the cells from mother. CONCLUSION: Through this study, we could conclude that several nucleated erythrocytes in maternal blood circulation can originate from mother, therefore we must develop the new method to identify the nucleated erythrocyte of fetal origin. Considering that we must apply for the larger number of pregnant women to screen, the procedure was multi-step and complex. Therefore, we must design the new scheme to utilize the nucleated erythrocytes in maternal blood.
Amelogenin ; Blood Circulation ; DNA ; Erythroblasts ; Erythrocytes ; Female ; Fetus ; Humans ; Male ; Mass Screening ; Micromanipulation ; Mothers ; Polymerase Chain Reaction ; Pregnancy ; Pregnant Women ; Y Chromosome

OBJECTIVE: We have introduced a method of characterization of invading glioma cells by using molecular analysis of marginal invading tumor cells and molecular profiles of glioma tumor margin. METHODS: Each of tumor core and marginal tissues was obtained in 22 glioma patients. Tumor core cells and marginal cells from each glial tumor were collected by laser capture microdissection or intraoperative microdissection under the operating microscope. Expression of MMP-2, MMP-9, CD44 and RHAMM mRNA by invading glioma cells compared with tumor core was confirmed by realtime-PCR of twenty-four glioma specimens. Clinical data also were reviewed for invasion and recurrence pattern of the gliomas radiologically and invasive rim pattern microscopically. RESULTS: Overall results of the molecular analysis showed that relative overexpression of MMP-2, MMP-9 and RHAMM were noted at the invasive edge of human glioma specimens comparing to the tumor core but CD44 was highly expressed in the tumor core comparing to the margin. High marginal expression of MMP-2 and MMP-9 were noted in poorly ill-defined margin on the pathological finding. High marginal expression of CD44 and MMP-2 were demonstrated in the midline cross group on the radiological review, and that of RHAMM and MMP-2 were showed in the aggressive recurrence group. High expression of MMP-2 seems to be involved in the various invasion-related phenomenons. CONCLUSION: Up-regulation of MMP-2, MMP-9, CD44 and RHAMM was noted in invasive edge of gliomas according to the various clinical situations.
Glioma* ; Humans ; Laser Capture Microdissection ; Microdissection ; Recurrence ; RNA, Messenger ; Up-Regulation

OBJECTIVE: This study was performed to determine the exact pattern of FHIT expression of the cervical carcinoma cell per se by microdissection and to investigate the clinical significance of the FHIT alteration in cervical cancer. METHODS: RT-PCR for FHIT transcript was performed in 18 cervical cancer tissues. Microdissection was performed using laser capture microdissection device and RNA was extracted by RT-nested PCR. PCR products were compared with known aberrant FHIT transcripts. Immunohistochemical analysis was performed to evaluate correlation between the altered expression of FHIT protein and clinical parameters. RESULTS: Six different size of aberrant FHIT transcripts were observed in cervical cancer tissues. Six of 18 (33.3%) cervical cancer sections exhibited full-length normal FHIT transcript only. Aberrant FHIT transcripts with normal one were observed in 9 (50%) and only aberrant transcripts in 3 (16.7%) frozen sections. Five normal cervical tissues expressed only a normal FHIT transcript. The sequences of the 6 different sizes of aberrant FHIT transcripts showed (1) deletion of exons 4-8, (2) deletion of exons 4-7, (3) deletion of exons 5-8, (4) deletion of exons 5-7, (5) deletion of exons 5-7 and insertion of intronic sequences, 153 bp, (6) deletion of exons 5-7 and insertion of intronic sequences, 84 bp. Microdissection of paired cervical tumor and normal stroma showed expression of aberrant FHIT transcripts only in tumor tissues. CONCLUSION: Aberrant FHIT expression was observed frequently in cervical carcinoma and they were observed mainly in cervical cancer cells by microdissection, but not in normal stromal cells. However, absence of FHIT expression did not correlate with clinical prognostic factors in cervical carcinoma.
Exons ; Frozen Sections ; Introns ; Laser Capture Microdissection ; Microdissection* ; Polymerase Chain Reaction ; RNA ; Stromal Cells ; Uterine Cervical Neoplasms*

Rapid progress has been made in the field of infertility since the first IVF (in vitro fertilization) baby was born in 1978. Controlled ovarian stimulation with FSH is currently the standard procedure for ovarian stimulation before follicular aspiration. Gonadotropin-releasing hormone agonists and antagonists have been used to prevent endogenous LH surge during controlled ovarian hyperstimulation.The goal of controlled ovarian stimulation with gonadotropins is to obtain a large number of mature oocytes and thereby improve the likelihood of obtaining an adequate number of embryos for subsequent transfer. IVF was initially presented as a treatment for tubal factor infertility but was quickly utilized in other areas in the field of infertility, such as male factor infertility and even ovarian failure. ICSI (intracytoplasmic sperm injection) is a more recent approach for male factor treatment, which allows the sperm to be directly injected into the egg using micromanipulation. Preimplantation genetic diagnosis can be performed on embryos prior to the embryo transfer. The complications associated with the IVF program include ovarian hyperstimulation syndrome and multiple pregnancies. The multiple pregnancies are directly related to the practice of transferring multiple embryos at embryo transfer. Each IVF clinic publishes its pregnancy rates. However, comparisons between clinics are difficult because the success rates vary depending on the distribution of underlying causes and age of the patients. The current take-home-baby rate is only 34.7%. In 2005, the Korean government enacted a law to regulate many aspects of IVF practice.
Embryo Transfer ; Embryonic Structures ; Female ; Fertilization in Vitro* ; Gonadotropin-Releasing Hormone ; Gonadotropins ; Humans ; Infertility ; Jurisprudence ; Male ; Micromanipulation ; Oocytes ; Ovarian Hyperstimulation Syndrome ; Ovulation Induction ; Ovum ; Pregnancy ; Pregnancy Rate ; Pregnancy, Multiple ; Preimplantation Diagnosis ; Reproductive Techniques, Assisted ; Sperm Injections, Intracytoplasmic ; Spermatozoa

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 (C57BLfemale symbol/CBAmale symbol). 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+/-14.0,) than in 6/8(26.5+/-6.2), 5/8(25.0+/-5.5), and 4/8(17.8+/-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+/-10.2), compared with the control (50.2+/-14.0), 7/8 (56.0+/-22.2), and 6/8 (55.3 +/-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.
Animals ; Biopsy ; Blastocyst ; Blastomeres ; Coculture Techniques* ; Cryopreservation ; Dimethyl Sulfoxide ; Embryonic Structures* ; Fertilization in Vitro ; Herpes Zoster ; Humans ; Mental Competency* ; Mice* ; Micromanipulation ; Preimplantation Diagnosis* ; Prostaglandins D ; Serum Albumin, Bovine ; Sucrose ; Survival Rate ; Vero Cells ; Zona Pellucida