The authors described the procedures for the establishment of clone 515 of the cell strain of osteochondrosarcoma by capillary method.A single cell was sucked into a capillary tube by a microsyringe under the control of a micromanipulator.A segment of the capillary tube with the cell was then planted into the plasma clot in a Carrel flask. After the addition of conditioned medium,the flask was incubated at 37℃.Daily ob- servations were made.The propagation of the single cell,the migration of its progenies out of the capillary tube and the change in form and size of the cells were recorded. 63 days after the isolation,the progenies covered the whole bottom of the flask.First subculture was then made and the clone was established therefrom.

Objective: To detect early subclinical signs of autonomic dysfunction in the cardiovascular system and explore the mechanism of orthostatic hypotension (OH) in patients with multiple system atrophy (MSA). Methods: Eighteen male patients with possible MSA and 10 healthy men were recruited. The hemodynamic responses to head-up tilt and tilt-reversal were studied by an electrically-powered tilt table and a non-invasive cardiac output measurement (NICOM) system. Results: At supine, there was no signifi cant difference in blood pressure, heart rate (HR), stroke volume, cardiac output and total peripheral resistance between MSA patients and healthy controls. During tilting upright, OH developed in 5 MSA patients, with a 23.7±4.8 mmHg drop in systolic blood pressure. Patients with OH were older and exhibited higher scores in unifi ed Multiple System Atrophy Rating Scale part I than patients without OH. The stroke volume, cardiac output and total peripheral resistance did not differ between groups. The controls had the most signifi cant HR elevation (6.5±2.5 bpm) during tiltup, followed by patients without OH (2.8±1.6 bpm) and those with OH (-0.2±2.2 bpm). A similar trend of HR decrease was observed during return to supine posture. The process of tilt-reversal altered HR more signifi cantly than head-up tilt in controls (8.0±2.9 vs 6.5±2.5 bpm; P=0.031) and patients without OH (4.2±2.1 vs 2.8±1.6 bpm; P=0.032), but not in patients with OH (1.2±1.5 vs -0.2±2.2 bpm; P=0.380). Conclusions: The HR change during postural challenge showed signifi cant difference between MSA patients and healthy controls. Impaired HR responsiveness contributed to OH in MSA. Monitoring HR during the tilt table test may be a practical and useful method to detect early autonomic dysfunction in patients with MSA.

PURPOSE: The liver is one of the most commonly injured organs in abdominal trauma. Surgery has played a major role in treating traumatic liver injury. Recently, it was reported that conservative treatment could be the first-line management for the hemodynamically stable patients without combined intraabdominal surgical problems. However, the risk of mortality remains high in this type of injury. The aim of this study was to examine the prognostic factors intraumatic liver injury. METHODS: The medical records of 132 patients who were treated for traumatic liver injury at Ewha Womans University, Mokdong Hospital from March 1994 to June 2001 were reviewed. The initial systolic blood pressure, prehospital transportation time, the number of associated organ injury, the level of initial hemoglobin, the grade of liver injury, the treatment method, the preoperative preparation time, the amount of estimated intraoperative blood loss, and the amount of intraoperative transfusion were analyzed with a univariate analysis and a multivariate analysis. RESULTS: Among the 132 patients, 76 cases (57.6%) were managed non-surgically, and 56 cases (42.4%) underwent surgery. Out of the 17 cases (12.9%) of mortality, 14 (82.4%) were encountered in those who underwent surgery and 3 cases (17.6%) were encountered in those who were treated nonsurgically. Univariate analysis revealed that the initial systolic blood pressure, prehospital transportation time, initial hemoglobin level, the number of associated organ injury, the surgical treatment, the grade of liver injury and the amount of intraoperative transfusion were significant prognostic factors for the survival. Multivariate analysis indicated that the initial systolic blood pressure and the extent of associated organ injury were the independent prognostic factors. CONCLUSION: The patients with unstable vital signs initially and multiple associated organ injuries have a poor prognosis. Aggressive management might be helpful for improving the survival rate in these patients.
Blood Pressure ; Female ; Humans ; Liver* ; Medical Records ; Mortality ; Multivariate Analysis ; Prognosis ; Survival Rate ; Transportation ; Vital Signs

Roundworms of Toxocara canis and Toxascaris leonina are common gastrointestinal helminths of canids over the world. Humans are infected with T. canis larvae through ingestion of infective eggs in contaminated environments or larvae by consumption of raw or uncooked meat or livers. Recently, patients of clinically diagnosed toxocariasis are increasing and require correct diagnosis in Korea. The present study investigated serological cross-reactivity between crude antigens of T. canis (TCLA) and T. leonina (TLLA) larvae. We collected serum specimens from 177 toxocariasis patients who were clinically suspected in the Seoul National University Hospital and 115 healthy controls. An ELISA method for toxocariasis was used to evaluate diagnostic efficacy of TLLA for serodiagnosis of human toxocariasis. The IgG ELISA using TLLA gave 14 (14.3%) positives of 98 TCLA positive specimens among 177 suspected toxocariasis patients. Most of them showed high absorbances with TCLA. In conclusion, there is a partial cross reaction between serum specimens of toxocariasis and TLLA.
Animals ; Antibodies, Helminth/blood ; Antigens, Helminth/*immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G/blood ; Larva/immunology/metabolism ; Toxascaris/growth & development/*immunology/isolation & purification ; Toxocara canis/growth & development/*immunology/isolation & purification ; Toxocariasis/*diagnosis/parasitology

The D-loop of mitochondrial DNA contain C tracts(homopoymeric cytosines) in the positions of 16183 -16193, 303 -315, 568 -573 of GI 1944628 of NCBI. When the sequence of these positions are analyzed, the sequencing reactions or their signals after the C tract were abruptly decreased and cannot clarify the nucleotide signals in many cases. It is hard to characterize of the C tract. To characterize the C tract, we tried to analyze the sequence of clone and amp-length polymorphism with silver stain after amplification of small fragments including C tract of 16183 -16193 and 303-315.These results were compared with the results of characterization of nucleotides after separation of each nucleotide signal from the electropherogram which were increased in the Y axis to intensify the small overlapped signals using CHROMAS program. All methods we tried showed similar pattern of heteroplasmies of homopolymeric cytosines. Conclusively, it is not necessary to try resequencing reaction after cloning or amp-length polymorphism using silver stain. Intensification of the small overlapped signals in Y axis and separation of each signal is enough method to characterize the C tract.
Axis, Cervical Vertebra ; Clone Cells ; Cloning, Organism ; DNA, Mitochondrial* ; Nucleotides ; Silver

Natural phytoestrogens such as the isoflavones genistein and daidzein, and the flavones quercetin exhibit anti-cancer properties. This study was purpose to investigate the anti-proliferative effect of phytoestrogens on acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) cells, and their synergistic antileukemic effect in combination with chemotherapeutic drugs. Optimal dosage of genistein, quercetin and in combination with chemicals for leukemia cells were determined by experiments. Cell viability, apoptosis induction and cell cycle arrest were detected by trypan blue staining, MTT assay, optical microscopy, flow cytometry (FCM). The schedule treatment of combination of genistein and chemicals was determined. The results showed that genistein exhibited a dose- and time-dependent inhibitory effect on cell proliferation in NB4 and HL-60 cells, induced apoptosis and cell cycle arrest in G2/M phase. Quercetin had evident inhibitory effect on the proliferation of K562 and K562/A cells. The combination of genistein and chemicals exerted a synergistic effect on cell growth inhibition. In conclusion, this study demonstrated the synergistic antileukemic effect of genistein with chemotherapeutic drugs on leukemic cells. This combination appears to be a new idea for the clinical novel treatment of leukemia.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Drug Synergism ; Genistein ; pharmacology ; HL-60 Cells ; Humans ; Isoflavones ; pharmacology ; Leukemia, Monocytic, Acute ; pathology ; Leukemia, Myeloid, Acute ; pathology ; Phytoestrogens ; pharmacology ; Quercetin ; pharmacology

OBJECTIVETo investigate the effects of hepatocyte growth factor (HGF) on adriamycin-induced apoptosis of gastric cancer cells in vitro.

METHODSAn eukaryotic expression plasmid (pIRES2-EGFP) containing HGF (pIRES2-EGFP-HGF) was constructed. Human gastric cancer cell line MKN-45 cells were transfected in vitro with pIRES2-EGFP containing HGF or not. RT-PCR and Western blot were used to determine the target gene expression. Function of HGF was determined by MDCK cell scattering assay. Cell viability was tested by MTT assay. Apoptosis was evaluated by DNA fragmentation assay as well as flow cytometry using PI staining.

RESULTSThe HGF transfected MKN45 cells could stably express HGF mRNA, and secrete HGF protein to the cell culture median which was detected to exhibit normal function. The cell inhibition rate induced by adriamycin in HGF-transfected cells was decreased as compared to that of parental and mock transfected cells. When treated with adriamycin at 0.1 microg/ml, the parental and mock transfected cells present typical apoptotic ladder on DNA electrophoresis while HGF transfected cell did not. The apoptotic rate was decreased in HGF transfected cells as compared with that of parental and mock transfected cells.

CONCLUSIONHGF gene transfection may suppress adriamycin-induced apoptosis of gastric cancer cells.

Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; genetics ; physiology ; DNA Fragmentation ; drug effects ; Dose-Response Relationship, Drug ; Doxorubicin ; pharmacology ; Flow Cytometry ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatocyte Growth Factor ; genetics ; metabolism ; physiology ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection

PURPOSE: There have been several recent reports of laparoscopy-assisted procedure for limited, or a laparoscopic, resections of small lesion of the liver. However, there are few reports on total laparoscopic surgery in anatomic resections, which is due to high level of skill required for the procedure and the risk of an air embolism. The aim of this study was to determine the safety of total laparoscopic surgery in anatomic liver resections during our early experiences. METHODS: The 6 patients included in this study were all treated with a totally laparoscopic anatomic resection of the liver at Ewha Womans University, Mokdong Hospital between September 2002 and January 2003. All 6 cases were diagnosed with an intrahepatic duct (IHD) stone, with stricture and/or common bile duct (CBD) stones. Of these 6 cases, 2 underwent a totally laparoscopic left lateral segmentectomy of the liver. In the remaining 4 cases, totally laparoscopic left lobectomies and CBD explorations, with T-tube insertion, were performed. In all cases, 4 trocars were used. During the operation, the intraperitoneal pressure of the pneumoperitoneum was maintained between 8 and 12 mmHg using CO2. The instruments used included Ligasure(R), Autosonix(R), 0 degrees and 30 degrees camera, fan retractor, Endo(vascular)- GIA 30 , and a large LapBag(R). All cases were managed in the supine position, with 15 degrees of left tilting. The specimen were extracted through a 4 to 5 cm extension of the upper trocar site. The T-tube exited through a 12mm site in the right upper port, and a drain was inserted into the sub-hepatic area. RESULTS: The patients comprised of 5 women and a man, with a mean age of 50.2 years old. The mean operative time was 469.2 (+/-141.4) minutes. In one case, 2 pints of packed red blood cells had to be transfused during the operation. The mean time to diet was 3.3 days. The mean postoperaitve hospital stay was 11.7 days. There was no conversion to open surgery, morbidity or mortality. The mean extensional incision size for the extraction of the specimen was 4.3 cm. CONCLUSION: In the anatomic resection of the liver, especially for benign liver disease, a total laparoscopic resection could be another relatively safe option for selected cases.
Common Bile Duct ; Constriction, Pathologic ; Conversion to Open Surgery ; Diet ; Embolism, Air ; Erythrocytes ; Female ; Humans ; Laparoscopy ; Length of Stay ; Liver Diseases ; Liver* ; Mastectomy, Segmental ; Mortality ; Operative Time ; Pneumoperitoneum ; Supine Position ; Surgical Instruments

The present authors investigated intestinal parasitic infections among North Korean residents and refugees in China in 2003. The Kato-Katz method was applied to 236 residents and soldiers in a town on the North Korea-China border and to 46 people at a refugee camp in China. Only eggs of Ascaris and Trichuris were detected, with egg positive rates of 41.1% and 37.6%, respectively. The total egg positive rate was 55.0% and most of those who were egg positive were only lightly infected. Women of 61.2% and men of 53.1% were egg positive. The refugees from rural areas showed higher egg positive rates than those from urban areas. The present investigation confirmed high prevalence of soil-transmitted intestinal helminths in rural borderline areas of North Korea.
Trichuris/*growth & development ; Trichuriasis/*epidemiology ; Male ; Korea/epidemiology ; Intestinal Diseases, Parasitic/*epidemiology/parasitology ; Humans ; Female ; Child ; Ascaris/*growth & development ; Ascariasis/*epidemiology ; Animals ; Adult ; Adolescent

We investigated a small-scale serological survey to screen tissue-parasitic helminthiases of North Koreans as one of research programs for re-unification of Korea. Soil-transmitted helminthiases were found highly prevalent among North Korean residents at the border with China. ELISA using 4 tissue-parasitic helminth antigens was applied to 137 residents living in Cheongjin-shi, Hamgyeongbuk-do, North Korea and 133 female refugees in South Korea in 2004-2005. Among a total of 270 samples, 31 (11.5%), 25 (9.3%), and 11 (4.1%) were positive for specific IgG antibodies to antigens of Clonorchis sinensis, Taenia solium metacestode, and sparganum, respectively. The overall positive rate was 21.5%; 38.2% in males and 15.8% in females. The present finding suggests that tissue parasites, such as C. sinensis, T. solium metacestode and sparganum are highly prevalent in some limited areas of North Korea. These foodborne tissue-parasitic helminthiases should be considered for future control measures of parasitic diseases in North Korea.
Adolescent ; Adult ; Aged ; Animals ; Antibodies, Helminth/blood ; Antigens, Helminth/blood ; Child ; Child, Preschool ; Clonorchis sinensis/isolation & purification ; Enzyme-Linked Immunosorbent Assay ; Female ; Helminthiasis/*epidemiology/*parasitology ; Humans ; Immunoglobulin G/blood ; Korea/epidemiology ; Male ; Middle Aged ; Prevalence ; Seroepidemiologic Studies ; Sparganum/isolation & purification ; Taenia solium/isolation & purification