OBJECTIVES: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. MATERIALS AND METHODS: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from LN2 and letting them at room temperature for 10 min followed by 37degrees C water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. RESULTS: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospermic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. CONCLUSIONS: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.
Baths ; Cryopreservation ; Fertilization ; Humans ; Insemination ; Linear Energy Transfer ; Pregnancy ; Pregnancy Rate ; Seminiferous Tubules ; Sperm Injections, Intracytoplasmic ; Spermatozoa* ; Water

Hereditary pancreatitis is a rare autosomal dominant inherited disease with 80% penetration rate. The disease is characterized by recurrent episodes of pancreatitis often beginning in childhood, positive family history with at least two other affected members and no known precipitating factors. Most forms of hereditary pancreatitis are caused by one of two commoner mutations, R122H in exon 3 and N29I in exon 2 of the cationic trypsinogen (CT) (PRSS1) gene, located on chromosome 7. These genetic defects are speculated to cause excessive trypsin activity or to prevent inactivation of prematurely activated trypsin, resulting in pancreatitis. We performed mutation analysis of a Korean family with two members having clinically suspicious hereditary pancreatitis. We analyzed the CT gene in DNA samples extracted from peripheral blood of five family members. First of all, polymerase chain reaction and restriction enzyme digestion were performed in exon 3 of the CT gene. And then DNA products were purified and sequenced. We found out that three members of the family, the mother and two daughters, had a R122H mutation of the CT gene. We report the first family of hereditary pancreatitis associated with the CT gene mutation, an arginine to histidine amino acid substitution at residue 122, in Korea.
Amino Acid Substitution ; Child ; DNA Mutational Analysis ; Female ; Gastrointestinal Hemorrhage/*etiology ; Humans ; Mutation ; Pancreatic Pseudocyst/*complications ; Pancreatitis/complications/*genetics ; Trypsinogen/*genetics

BACKGROUND: Metabotropic glutamate receptors (mGluRs) participate in the induction of synaptic plasticity phenomena, such as long-term potentiation and long-term depression that are thought to be at the origin of learning and memory. They are also likely to play a role in modulating glutamate-induced neurotoxicity. It will become apparent that mGluRs are excellent targets for the development of drugs that modulate excitatory synaptic transmission. But there were several controversies about the exact role of group 1 mGluRs subtype 5 (mGluR5). This study was designed for evaluation of the neuroprotective role of mGluR5. METHODS: Fifty male Sprague-Dawley rats were divided into three groups, control, MK-801 and lamotrigine. The hippocampus and basal ganglia were removed at 6 hours and 3 days after the one hour transient middle cerebral artery occlusion. The gene expression of mRNA of the brain samples were evaluated by using reverse transcriptase polymerase chain reaction technique. RESULTS: The gene expression of mGluR5 mRNA in hippocampus was increased by 101.96 +/- 18.45% at 6 hours after ischemia and decreased by 50.70 +/- 15.73% at 3 days after ischemia (p<0.01). MK-801 and lamotrigine attenuated the ischemia-induced increases of gene expression of mGluR5 mRNA. In MK-801 group, the expression in basal ganglia was increased by only 0.23 +/- 5.41% at 6 hours after ischemia and decreased by 9.82 +/- 4.35% at 3 days after ischemia. In MK-801 group, the expression in hippocampus was decreased by 3.45 +/- 8.24% and 9.35 5.69% at 6 hours and 3 days after ischemia. In lamotrigine group, the expressions in hippocampus and basal ganglia were decreased by 26.66 +/- 9.85% and 9.45 +/- 5.22% at 6 hours after ischemia. CONCLUSIONS: From these results, the role of mGluR5 was defined as a mediator for neuronal damage after transient focal cerebral ischemia in hippocampus and basal ganglia.
Animals ; Basal Ganglia ; Brain Ischemia* ; Brain* ; Control Groups ; Depression ; Dizocilpine Maleate ; Gene Expression ; Glutamic Acid* ; Hippocampus* ; Humans ; Infarction, Middle Cerebral Artery ; Ischemia ; Learning ; Long-Term Potentiation ; Male ; Memory ; Neurons ; Plastics ; Rats* ; Rats, Sprague-Dawley ; Receptors, Glutamate* ; Receptors, Metabotropic Glutamate ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Synaptic Transmission

No abstract available.
Humans* ; Pregnancy* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa*

The combination of testicular sperm extraction (TESE) with ICSI can achieve normal fertilization and pregnancy rate and is affective method in obstructive and non-obstructive azoospermic patients. But, when pregnancy was not occurred, repeated testicular biopsies are not evitable. Therefore, in this study, we observed the survival rate of testicular spermatozoa and spermatozoa extracted from the seminiferous tubules after cryopreserved-thawed used for next IVF cycle with ICSI. In a total of 23 cases, obstructive azoospermia was 17 cases and non-obstructive azoospermia was 6 cases. In obstructive azoospermia, after thawing, motile spermatozoa was observed in 13 cases (76.5%). The fertilization rate with 2PN was 67.6% and 5 pregnancies (29.4%) were achieved. In non-obstructive azoospermia, motile spermatozoa was observed in 2 case (33.3%) after thawing. The fertilization rates with 2PN was 53.7% and 3 pregnancies (50.0%) were achieved. A comparison of the results of motile spermatozoa after thawed testicular spermatozoa find spermatozoa extracted from the thawed seminiferous tubule section were 3 cases (60.0%) and 12 cases (86.6%), respectively, The fertilization and pregnancy rates of thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 69.4% and 20.0%, 62.5% and 38.8%, respectively. Conclusively, thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section can achieve normal fertilization and pregnancy and cryopreservation of testicular spermatozoa and seminiferous tubule may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testis.
Azoospermia ; Biopsy ; Cryopreservation ; Fertilization ; Humans ; Pregnancy ; Pregnancy Rate ; Seminiferous Tubules* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa* ; Survival Rate ; Testis

OBJECTIVES: Klinefelter syndrome is the most common genetic cause of male infertility and presents with 47, XXY mainly or 46, XX/47, XXY mosaicism. It is characterized by hypogonadism and azoospermia due to testicular failure, however, sporadic cases of natural pregnancies have been reported. With the development of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), sperm can be retrieved successfully and ART is applied in these patients for pregnancy. It has been suggested that the risk of chromosome aneuploidy for both sex chromosome and autosome is increased in the sperms from 47, XXY germ cells. Considering the risk for chromosomal aneuploidy in the offspring, preimplantation genetic diagnosis (PGD) could be applied as a safe and more effective treatment option in Klinefelter syndrome. The aim of this study is to assess the outcome of PGD cycles by using FISH for sex chromosome and autosome in patients with Klinefelter syndrome. MATERIALS AND METHODS: From Jan. 2001 to Dec. 2003, PGD was attempted in 8 cases of Klinefelter syndrome but TESE was failed to retrieve sperm in the 3 cases, therefore PGD was performed in 8 cycles of 5 cases (four 47, XXY and one 46, XY/47, XXY mosaicism). In one case, ejaculated sperm was used and in 4 cases, TESE sperm was used for ICSI. After fertilization, blastomere biopsy was performed in 6~10 cell stage embryo and the chromosome aneuploidy was diagnosed by using FISH with CEP probes for chromosome X, Y and 17 or 18. RESULTS: A total of 127 oocytes were retrieved and ICSI was performed in 113 mature oocytes. The fertilization rate was 65.3+/-6.0% (mean+/-SEM) and 76 embryos were obtained. Blastomere biopsy was performed in 61 developing embryos and FISH analysis was successful in 95.1% of the biopsied blastomeres (58/61). The rate of balanced embryos for chromosome X, Y and 17 or 18 was 39.7+/-6.9%. The rate of aneuploidy for sex chromosome (X and Y) was 45.9+/-5.3% and 43.2+/-5.8% for chromosome 17 or 18, respectively. Embryo transfer was performed in all 8 cycles and mean number of transferred embryos was 2.5+/-0.5. In 2 cases, clinical pregnancies were obtained and normal 46, XX and 46, XY karyotypes were confirmed by amniocentesis, respectively. Healthy male and female babies were delivered uneventfully at term. CONCLUSION: The patients with Klinefelter syndrome can benefit from ART with TESE and ICSI. Considering the risk of aneuploidy for both sex chromosome and autosome in the sperms and embryos of Klinefelter syndrome, PGD could be offered as safe and more effective treatment option.
Amniocentesis ; Aneuploidy ; Azoospermia ; Biopsy ; Blastomeres ; Chromosomes, Human, Pair 17 ; Embryo Transfer ; Embryonic Structures ; Female ; Fertilization ; Germ Cells ; Humans ; Hypogonadism ; Infertility, Male ; Karyotype ; Klinefelter Syndrome* ; Male ; Mosaicism ; Oocytes ; Pregnancy ; Preimplantation Diagnosis* ; Prostaglandins D ; Sex Chromosomes ; Sperm Injections, Intracytoplasmic ; Spermatozoa

This study was carried to determine the possibility of finding motile spermatozoa and fertilization, pregnancy rate after testicular sperm extraction(TESE) with ICSI in obstructive and non-obstructive azoospermic patients. In 154 cases(132 patients), obstructive azoospermia was 77 cases and non-obstructive azoospermia was 77 cases. In obstructive azoospermia, patients generally showed normal spermatogenesis and included vas agenesis(n=8), multiple vas obstruction(n=7), epididymal obstruction (n=54). Total of 982 retrieved oocytes were obtained and 84.4% were injected. The fertilization rates with 2 PN and cleavage rate were 72.5% and 62.3%, .respectively. 30 pregnancies(38.9%) were achieved and the ongoing pregnancies were 22 cases (28.6%). In non-obstructive azoospermia, patients showed hypospermatogenesis(n=49), maturation arrest(n=4), Sertoli cell only syndrome (n=24). The various stages of spermatogenic cell could be retrieved by TESE and could be reached normal fertilization and embryo development with ICSI. Total of 1072 retrieved oocytes obtained and 80.2% were injected. The fertilization rates with 2 PN and cleavage rate were 52.8% and 68.9%, respectively. 22 pregnancies(30.1%) were achieved and the ongoing pregnancies were 19 cases(26.0%). Conclusively, the combination of TESE with ICSI using testicular spermatozoa can achieve normal fertilization and pregnancy rate and effective method in obstructive and non-obstructive azoospermic patients.
Azoospermia ; Embryonic Development ; Female ; Fertilization* ; Humans ; Oocytes ; Pregnancy Rate* ; Pregnancy* ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Spermatogenesis ; Spermatozoa*

Irreparable obstructive azoospermic patients can be treated successfully with microsurgical epididymal sperm aspiration(MESA) o. testicular sperm extraction (TESE) by intracytoplasmic sperm injection(ICSI). Obstructive azoospermic patients generally have normal spermatogenesis. The aim of this study was to see if any spermatozoa could be retrieved from non-obstructive azoospermia and to assess the efficacy of ICSI with TESE in germinal failure. 42 non-obstructive azoospermic patients revealed no spermatozoa at all in their ejaculates, even after centrifuge. The histology of 42 patients revealed 15 Sertoli cell only Syndrome, 4 maturation arrest and 23 severe hypospermatogenesis. All patients underwent extensive multiple testicular biopsy for sperm retrieval. These patients were scheduled for ICSI using testicular spermatozoa. In 25 out of 42 non-obstructive azoospermic patients, spermatozoa were recovered from multiple testicular biopsy specimen and 11 ongoing pregnancies were achieved. There are usually some tiny foci of spermatogenesis which allow TESE with ICSI in non-obstructive azoospermia. Also these patients may have sufficient sperm in the testes for ICSI, despite extremely high FSH level and small testes.
Azoospermia* ; Biopsy ; Humans ; Male ; Oligospermia ; Pregnancy ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatogenesis ; Spermatozoa ; Testis

PURPOSE: The supply of donor organs does not meet the demand of transplantation. To find a possible way of expanding the donor pool, we have evaluated the risk factors of cadaveric donors that influence the graft survival. METHODS: Between Feb. 1995 and Dec. 1999, we performed 200 cases of cadaveric kidney transplantation. The graft survival rates in 1 yr, 2 yr, & 3 yr are 93.0%, 91.1%, & 89.9%, respectively. We defined the marginal donor as a donor who possesses one of the risk factors of donor age > or = 50, or < or =5 year old (n=18), ICU stay > or =10 days (n=46), history of hypertension (n=30), CPR > or =20 min (n=7), prolonged hypotension (SBP < or =80 mmHg, over 6 hr) (n=40), high dose inotropic support (dopamine > or =20microgram/kg/min, or dobutamine > or =15microgram/kg/min) (n=78), serum creatinine level > or =2.5 mg/dl (n=16), or cold ischemic time > or =12 hr (n=34). We compared graft survival rates between the marginal donor group and the non-marginal donor group, and analyzed risk factors affecting graft survival by univariate and multivariate methods. RESULTS: CPR > or =20 min (71.5%, 71.5%, - vs 93.8%, 91.8%, 90.6%, p=0.027), prolonged hypotension (85.0%, 85.0%, 80.3% vs 95.0%, 92.6%, 92.6%, p=0.028) and serum creatinine > or =2.5 mg/dl (75.0%, 75.0%, 75.0% vs 94.6%, 92.4%, 91.0%, p=0.001) affect graft survival in univariate analysis. There is, However, no risk factor that affects graft survival in multivariate analysis. CONCLUSION: We can increase the supply of donor organs for kidney transplantation by the selective use of high-risk cadaver donor. However, a prospective randomized study including recipient factors is needed to define the acceptable conditions better.
Risk Factors

Purpose: Currently, trastuzumab plus chemotherapy is the standard first-line therapy for human epidermal growth factor receptor 2 (HER2)-positive advanced or metastatic gastric cancer (mGC) or esophagogastric junction cancer. However, it is not clear whether the prognosis of HER2-positive mGC treated with trastuzumab plus chemotherapy is better than that of HER2-negative mGC treated with chemotherapy as the first-line therapy. Methods: We performed a retrospective study comparing the prognosis of mGC according to first-line treatment with trastuzumab plus chemotherapy or chemotherapy only, at the Korea Cancer Center Hospital from 2011 to 2018. The Kaplan-Meier method and Cox proportional hazards model were used for univariate and multivariate survival analyses. Results: The median overall survival of trastuzumab group was 26.1 months and that of chemotherapy group was 14.8 months (P=0.047). Trastuzumab group had a longer median progression-free survival than chemotherapy group (23.4 vs. 9.2 months, P=0.026). By univariate analysis, sex, age, World Health Organization (WHO) histology, HER2 status, primary tumor site, extent of disease, number of lesions, number of metastatic, measurability of disease, prior gastrectomy, and chemotherapy group are statistically significant. Using multivariate analysis, number of lesions, number of metastatic, prior gastrectomy, and trastuzumab group (hazard ratio, 0.594; 95% confidence interval, 0.384–0.921; P=0.020) were found to be independent prognostic factors of overall survival. Conclusion The result suggests prognosis of HER2-positive mGC treated by trastuzumab plus chemotherapy could be better than that of HER2-negative mGC treated by chemotherapy only. Well-designed prospective cohort studies are needed to confirm the results of this study. HER2 testing should be performed routinely in all patients newly diagnosed with mGC.