The effects of leptin on in vitro development of mouse embryos / 대한산부인과학회지

OBJECTIVE: To investigate the effects of leptin on in vitro development of mouse embryos. METHODS: Female C57BL mice, aged 8 to 15 weeks, were superovulated with IP injection of 5 IU of PMSG followed by 5 IU hCG at 48 hours later. And then the mice were mated with male mice. The next morning, one-cell embryos were collected and cultured in media added with various concentrations (0, 5, 50, 500 ng/mL) of leptin for 4 days. In addition, to determine whether the sensitivity to leptin varied at different stages of development, embryos at 2- or 4-cell stage were treated with the same concentrations of leptin and cultured up to blastocyst stage. The total cell number of blastocyst was assessed and the expression of leptin receptor was examined in all stages of development by immnuofluorescence. RESULTS: The rate of blastocyst formation from one-cell embryos significantly increased at culture media that leptin was added at 50 ng/mL concentration, whereas decreased at 500 ng/mL concentration compared to the control (P<0.05). The development rate of embryos, from 2-cell stage, was similar to the rate from 1-cell stage. However, the addition of leptin to culture media in 4-cell embryos had no significant effects on embryo development compared to the control. In addition, the dose-dependent stimulatory or inhibitory effect of leptin on embryo development was weakened at 2-cell and 4-cell embryo stages compared one-cell embryo stage. The total cell number of blastocyst also significantly increased at 50 ng/mL of leptin, but decreased at 500 ng/mL. Leptin receptor was expressed in all stages from one-cell embryos to blastocyst. The intensity of Ob-Rb immnuostaining was mainly stronger in one- or two-cell embryos, decreased with advancing development stages, and increased again in blastocyst. CONCLUSION: This study shows that addition of leptin to embryo culture media affects embryo development in a dose-dependent and developmental stage-dependent manner. The effects of leptin seems to be associated with the expression pattern of leptin receptor at different stages of development.

Interstitial pregnancies treated by endoscopic cornuotomy after ingection of large amount of diluted vasopressin / 대한산부인과학회지

OBJECTIVE: Our aim was to report an effective procedure of injection of large amount of diluted vasopressin in the treatment of interstitial pregnancy using endoscopic cornuotomy METHODS: This is an uncontrolled retrospective study of 17 patients interstitial pregnancies treated of by endoscopic cornuotomy from June 2001 to June 2004 in our hospital. All cases were applied by the large amount of diluted vasopressin injection. RESULTS: Among 446 ectopic pregnancies, interstitial pregnancies were 17 (3.81%). Fourteen patients (82.4%) were unruptured, three patients (17.6%) were ruptured. Mean (+/-SD) age and parity of patients were 35.6+/-3.9 years (range 30-40 years), 1.5+/-2.1 (range 0-3), respectively. At the time of operation the duration of amenorrhea and mean size of ectopic gestation were 54.5+/-9.6 day and 2.6 x 2.6 cm, respectively. The amount of injected vasopressin was 190.0+/-55.5 cc. For the cornual closure, the endosuture method was applied in 10 patients (59%), and the endoloop method was applied in 7 patients (41%) The mean (+/-SD) blood loss and operation time were 18.2+/-15.9 cc and 28.3+/-7.8 minute, respectively. In fifteen patients (88%), serum beta-hCG level from the preoperative base was declined 90% or more within postoperative day 4 and in 2 patients (12%), it was decreased within postoperative day 60. CONCLUSION: Endoscopic cornuotomy using the large amount of diluted vasopressin injection procedure is a safe and effective in the interstitial pregnancy.

Successful endoscopic operations in benign gynecologic diseases: experiences of 6,017 cases / 대한산부인과학회지

OBJECTIVE: The purpose of this study is to evaluate the application rate and effectiveness of endoscopic operations based on 6,017 cases in the field of gynecology. METHODS: Six-thousand-seventeen cases of endoscopic operations were performed from March 1993 to June 2004 in the department of Obstetrics and Gynecology, Center for Endoscopy, Good Moonhwa Hospital. Endoscopic operations were done on the majority of patients who had benign gynecologic diseases. We applied "two-surgeons system" and simplified methods of operation consisted of endoloop-ligation, endosuture and endotying. RESULTS: Our endoscopic operations were successful in 5,994 cases (99.62%) out of 6,017 attempts. Twenty-three cases (0.38%) were converted to laparotomy due to severe pelvic adhesion, excessive bleeding or difficulty in accessibility. In the first year endoscopic operations were done in 29.0% of the total gynecologic operations, but was increased to 52.9% during the next 4 years. In 2003, 93.0% and in 2004, 99.0% of the total gynecologic operations were done by endoscope. While the simple technique surgeries showed a high application rate, the more complicated technique surgeries, which began at a lower rate, showed a steady increase up to 89.3%. Blood loss and operation time were markedly declined during 3 year period and maintained at a low level thereafter. A 3 years' period of endoscopic operations (about 500 cases) is necessary for a hospital learning curve while, 26.4 cases is necessary an individual surgeon's learning curve. The major complications during the study period was 28 cases (0.47%). CONCLUSION: Endoscopic operations can be applied in almost all benign gynecologic diseases. In particular, "two-surgeons system" is a safe method with few complications and minimal learning period. These results suggest that if appropriate techniques could be learned, endoscopic operation might be preferred to laparotomy for both surgeons and patients.

Health Effects of Air Particle Produced in a Waste Treatment Plant on Residents Living in an Official Residence of Air Base

BACKGROUND: This study was conducted to reveal the health effects of air pollution on the residents living near waste treatment plant. METHODS: We sampled 190 residents as the hogh exposed group, 192 residents as the moderate group, and 139 residents as the low exposed group according to the geographical distances from the waste treatment plant. Self-administered questionnaires for prevalence of respiratory disease, irritation symptoms of mucous membrane (eye, nose, thoat, skin, and respiratory tract), gastrointestinal symptoms, other nonspecific symptoms were conducted. We measured the concentrations of total suspened partcles, Zn, Cu, Cr, Ni, Pb, Cd and As in 3 exposed area. RESULT: Airborne concentrations of total suspended particles measured in exposed area were 485, 365, and 332 micro gram/m(3) which excees the limit value. Airborne concentrations of Zn, Cu, Cr, Ni, Pb, Cd and As were not excees the limit value. The prevalence of respiratory disease of high exposed group (16.8%) was significantly higher than that of moderate group (3.6%) and low exposed group (5.0%). The sysptom score of irrtation sysptoms of mucous membrane, gastrointestinal sysptoms, and other nonspecific sysptoms were increased with exposed level. In multiple linear regression, exposure level was related to all sysptom scores significantly. CONCLUSION: These results suggest that the air pollution of waste treatment plant would influence prevalence of respiratory disease, induce irritation of mucous membrane, gastrointestinal symptoms, and other nonspecific sysptoms.

Fine Needle Aspiration Cytologic Features of Schwannomas / 대한세포병리학회지

Schwannoma is a benign encapsulated nerve sheath tumor and is histologically characterized by a mixture of Antoni type A and B tissue. A preoperative diagnosis of schwannoma by fine needle aspiration cytology or by any other means is important to preserve clinically important nerves. Therefore, it is necessary to make a specific cytological diagnosis of nerve sheath tumor. However, there are a few reports regarding cytological features of schwannoma in Korea. We experienced seven cases of solitary schwannoma and here we report their characteristic cytological features with a review of literatures. The characteristic features of schwannoma on cytology were the presence of both Antoni type A and B tissue. The cytologic findings common to all cases of schwannoma generally corresponded to the histologic findings of Antoni type A tissue, consisting of fragments of tightly cohesive fascicles with variable cellularity. Dense fibrillary substances were found, along with palisading nuclei and Verocay bodies. Individual tumor cells consisted of cohesive cells having spindle or oval nuclei, with pointed ends and indistinct cell borders. Variation in nuclear size and shape was also present. The Antoni type B consisted of scattered wavy or short spindle cells and some histiocytes and lymphocytes in the abundant myxoid background with formation of microcysts. Immunohistochemistry for S-100 protein revealed a uniformly strong positive reaction and was helpful to make more accurate diagnosis of schwannoma.

Expression of bcl-2 and p53 Protein in Primary Gastric Lymphomas

The bcl-2 gene is a proto-oncogene which extends cell survival by blocking apoptosis. Bcl-2 expression has been detected in many types of nodal and MALT lymphoma. The p53 gene is a tumor suppressor gene and p53 mutation is the most common genetic alteration in human malignancies. The relationship between the expression of bcl-2 and p53 protein in primary gastric lymphoma has been rarely reported. The authors investigated the expression of bcl-2 and p53 protein in 37 cases of primary gastric lymphoma by immunohistochemical method using bcl-2 and p53 monoclonal antibodies. There were five cases of low grade B-cell MALT lymphomas and thirty two cases of high grade B-cell lymphomas. Fifteen of 37 cases (41%) showed bcl-2 protein expression in the cytoplasm of tumor cells and 26 cases (70%) showed p53 protein expression in the nucleus of tumor cells. Bcl-2 protein was detected in 4 of 5 (80%) low grade MALT lymphomas, and in 11 of 32 (34%) high grade lymphomas. There was no significant correlation between bcl-2 expression and histologic grade of primary gastric lymphomas (p>0.05). p53 protein was positive in 25 of 32 (78%) high grade lymphomas, and in 1 of 5 (20%) low grade MALT lymphomas. The expression of p53 protein is significantly higher in high grade lymphoma than in low grade MALT lymphoma (p<0.05). The p53 expression in the bcl-2 negative cases (86%) was significantly higher than in the bcl-2 positive cases (47%). There was an inverse relationship between bcl-2 and p53 expression in primary gastric lymphoma. These results suggest that bcl-2 and p53 expression in primary gastric lymphoma may be involved in the transition from low grade MALT lymphoma to high grade lymphoma.

Synaptic Reorganization of Dentate Mossy Fibers and Expression of Calcium Binding Proteins in Hippocampal Sclerosis of Temporal Lobe Epilepsy

This study was designed to identify expression of calcium-binding proteins and synaptic reorganizations of dentate mossy fibers in hippocampal sclerosis of human temporal lobe epilepsy. Hippocampal neuronal density was quantitively analyzed in temporal lobe epilepsy group (n=50) to investigate the degree of hippocampal sclerosis and it was compared with that of autopsy control (n=3). To verify the distribution of calcium-binding proteins in neurons of epileptic hippocampi, the parvalbumin (PV)-immunoreactive and calbindin-D28K (CB)-immunoreactive neurons were quantitively analyzed in each area of Ammon's horn by immunohistochemical stain. Also, to clarify synaptic reorganizations of the dentate mossy fibers, a part of each hippocampus was examined under light microscopy and transmission electron microscopy using Timm sulphide silver method. In epileptic hippocampi, severity of hippocampal sclerosis (HS) was graded four, which consisted of 3 cases with no HS, 6 mild HS, 12 moderate HS, and 29 severe HS. The hippocampal neuronal loss was most prominent in CA1, followed by CA4 and CA2. Expression of calcium-binding proteins was more prevalent in CA2 of all groups. The proportion of PV-immunoreactive neurons in CA1 and CA4 significantly increased in the moderate and severe HS group, whereas the proportion of CB-immunoreactive neurons did not correlated with the severity of HS. Timm granules were noted in inner molecular supragranular layer of dentate gyrus of epileptic hippocampi and they tended to increase in proportion along with the severity of hippocampal sclerosis. Transmission electron microscopy showed that supragranular Timm granules corresponded to synaptic terminals of mossy fibers. These results suggest that parvalbumin appears to have more protective effect against neuronal loss and that mossy fiber synaptic reorganization seems to play a major role in pathogenesis of hippocampal sclerosis of human temporal lobe epilepsy.

Cytokine Expression on Microglial Proliferation and Apoptosis in Rat Lumbar Spinal Cord Following Unilateral Sciatic Nerve Transection

This study was carried out to elucidate the cytokine mRNAs expression and morphological features according to a microglial proliferation and apoptosis in a rat lumbar spinal cord, after a right sciatic nerve transection. The control group was composed of 5 rats (Spraque-Dawley) and the experimental group was composed of 70 rats. On post operation day (pod) 1, 2, 3, 5, and 7 eight rats were sacrificed on those days. On pod 10 five rats were sacrificed as well as five rats sacrificed on post operation weeks 2, 3, 4, 5, and 6. On light microscopy, activated microglia were often found in a perineuronal position around motoneurons in the ventral gray matter and more randomly distributed throughout the neuropil in the dorsal gray matter of lumbar spinal cord. GSA I-B4-positive microglia began to increase from 1 day after transection, and reached peak at 2~3 days and it persisted at 5~7 days and decreased thereafter. TUNEL-positive microglia was not observed in control group and began to increase from 5 days after transection and increased gradually until 3 weeks and decreased thereafter. On in situ RT-PCR, the positive signal for IL-1alpha and IL-6 mRNA was found mainly in the cytoplasm of the activated microglia and astrocytes at 1 day after transection and showed stronger signal at 3 days and decreased gradually until 10 days. TNF-alpha mRNA was detected 1 day after transection and remained for 7 days and localized to activated microglia as well as probably some astrocytes. The signal intensity of IL-1alpha and IL-6 mRNA was generally stronger than TNF-alpha mRNA. On transmission electron microscopy, there were chromatin condensation with margination toward nuclear membrane and condensation of cytoplasm at 3 days after transection. Apoptotic bodies were found after 5 days and increased gradually until 3 weeks. According to the above findings, it is concluded that apoptosis appears to be one mechanism by which activated microglia are gradually eliminated and cytokine expression seems to played an active role in the microglial turnover.

Immunohistochemical Detection of p53 Gene Mutation in Urine Samples in the Patients with Bladder Cancer / 대한세포병리학회지

Although bladder cancers are very common, little is known about their molecular pathogenesis. It is known, that p53 alteration is found in about 60%p of muscleinvasive bladder cancer, necessiating aggressive therapy and poor outcome. We examined the nuclear expression of p53 protein, using D07 monoclonal antibody in the urine samples, from 31 patients with transitional cell carcinoma of the bladder to investigate the correlation of p53 overexpression with histologic grades and depth of invasion. The positive rate of p53 protein was 27%o in superficial bladder tumor, but increased up to 71% in the invasive bladder carcinomas. The overexpression of p53 protein increased according to Mostofi grading system from 18% in grade I, 45% in grade Il, and up to 100% in grade ill. The p53 expression tended to be higher in the invasive and high grade bladder cancers than in the superficial and low grade ones(p<0.05). These results suggest that immunohistochemical analysis of the urine specimen in the bladder cancer patients could be a useful method of screening for the presence of p53 mutant protein. The mutant p53 protein expression may be an indicator of bladder cancer with more proliferative potential and/or aggressive biologic behavior.

Diagnostic Application of p53 IMMUNOSTAINING in Bronchial Brush Specimens / 대한세포병리학회지

Abnormalities of p53 gene are common in lung cancers and are associated with immunologically detectable p53 protein. p53 immunoreactivity is uncommon in normal cells but is frequently seen in neoplasia. Therefore, assessment of p53 expression may assist in the cytological diagnosis of malignancy. The usefulness of p53 immunostaining as a marker of malignancy in the cytological analysis of bronchial brush specimens from the patients with lung cancers was investigated in this study. A total of 71 bronchial brush samples submitted for cytologic diagnosis were immunostained with D07, a monoclonal antibody to recombinant p53 protein. Resultant p53 data were correlated with cytologic diagnosis and clinical information. Of the 17 smears with a benign cytodiagnosis, all were p53 negative. Of the 40 cases with a malignant cytodiagnosis(histologically confirmed), 35 were p53 positive and 5 were negative. Of the 14 cases that were cytologically suspicious but nondiagnostic for malignancy, 11 were p53 positive, 9 of which were subsequently proved to be malignant by histologic examination, and the remaining 2 cases were tuberculosis clinically. Forty four of 51 histologically confirmed lung carcinomas were p53 positive, including 25 of 28 squamous cell carcinomas, 13 of 17 small cell carcinomas, 3 of 3 adeno- carcinomas, and 3 of 3 large cell undifferentiated carcinomas. These results suggest that p53 immunostaining could be of value as a marker of malignancy in the cytologic examination of bronchial brush specimens. Furthermore, we have shown the possible clinical utility of p53 immunostaining in cytopathological diagnosis, that is, as a valuable adjunct to morphological assessment in the analysis of cytopathologically suspicious cases.