BACKGROUND: Human cells are almost never spontaneously immortalized in vitro. We tried to immortalize human fetal hepatocytes (h-FH) and evaluate the differentiational status and its change. METHODS: Hepatocytes were isolated from a liver fragment of 20 week old fetus and infected with amphotropic recombinant retrovirus containing a temperature- sensitive mutant of SV40 large T antigen and neomycin phosphotransferase gene. G418 resistant colonies were cloned and expanded. The cells which were able to divide more than 30 times were used to analyze various functions. RESULTS: The immortalization rate was 3.3 x 10-8 and two cell lines (C11, D21) were established. C11-60, C11-80, D21-30 and D21-60 (suffix number means the cell division counts) were evaluated. D21-30 was thougt to be imcompletely immortalized because a considerable portion of cells died during culture. The morphology was similar to that of epithelial cells except for D21-30 which looked like fibroblast. The cells grew rapidly at 33oC but stopped growing at 39oC. T antigen and p53 was expressed at 33oC but disappeared at 39oC, which suggest that T antigen binds to p53. Chromosomal changes were so marked that it was impossible to discriminate exact number. Albumin was secreted as about 1/10 as that of h-FH, but alpha-fetoprotein secretion stopped after immortalization. Telomerase was activated in both cell lines except for the incompletely immortalized cells D21-30. Telomere was elongated in competely immortalized cell lines, but it was rather shortened in D21-30 compared to that of h-FH. Macroscopic colonies did not develop in soft agar assay. CONCLUSIONS: We successfully immortalized human fetal hepatocytes. Although the cells are not likely to have oncogenicity, the functions are not so good, possibly due to marked chromosomal changes which are thought to occur before telomerase is activated during immortalization step.
Agar ; alpha-Fetoproteins ; Antigens, Viral, Tumor* ; Cell Division ; Cell Line ; Clone Cells ; Epithelial Cells ; Fetus ; Fibroblasts ; Hepatocytes* ; Humans* ; Kanamycin Kinase ; Liver ; Retroviridae* ; Telomerase ; Telomere

We report a case of ring chromosome 13 with a distal deletion of 13q32.2-->qter observed in a fetus who was referred to our institution at term due to severe growth restriction and multiple congenital malformations on ultrasonographic examination.This boy was born by vaginal delivery at 39 weeks in gestation. His weight, head circumference and height were less than the 3 percentile of gestational age. Apgar score was 7 at 1 minute and 9 at 5 minutes. He showed microcephaly, large forehead, low set ears, hypertelorism, flat nasal bridge, and micrognathia. The genitalia was ambiguous, showing severe hypoplasia of the penis. The anus was ectopic, displaced anteriorly from its normal position but with a normal opening and function. Neurologic examination was normal. Echocardiogram done at 2 weeks of life showed a persistent foramen ovale and a ventriculoseptal defect (type II) with increased pulmonary hypertension. MRI examination of the brain showed poorly demarcating corpus callosum suspecting agenesis of corpus callosum. Also, cerebellar vermis was small and hypoplastic, mimicking a variant form of Dandy-Walker malformation. MRI of the pelvis showed a tubular structure in pelvic cavity, suspicious of uterine remnant, between urinary bladder and rectum, and a inguinal hernia was noted in the left side. In the abdominal cavity enlarged adrenal glands were noted, and hormonal study showed elevated 17-alpha-OH-progesterone (168.9 ng/ml) with normal 17-KS and 17-OHCS levels. Gastrointestinal and urogenital system were otherwise normal. Cytogenetic analysis of the parents were both normal but the newborn showed 46, XY, r (13), de novo, with deletion points q32.2-->qter. Our findings are in line with previous reports about chromosome 13 deletions, in which loss of the "critical point" leads to major malformations like brain anomalies and ambiguous genitalia.
Abdominal Cavity ; Adrenal Glands ; Agenesis of Corpus Callosum ; Anal Canal ; Apgar Score ; Brain ; Chromosomes, Human, Pair 13 ; Corpus Callosum ; Cytogenetic Analysis ; Dandy-Walker Syndrome ; Disorders of Sex Development ; Ear ; Fetus ; Foramen Ovale ; Forehead ; Genitalia ; Gestational Age ; Head ; Hernia, Inguinal ; Humans ; Hypertelorism ; Hypertension, Pulmonary ; Infant, Newborn ; Magnetic Resonance Imaging ; Male ; Microcephaly ; Neurologic Examination ; Parents ; Pelvis ; Penis ; Pregnancy ; Rectum ; Ring Chromosomes ; Urinary Bladder ; Urogenital System

OBJECTIVE: The purpose of study was to assess in a longitudinal study of maternal plasma concentrations of Tumor necrosis factor-alpha(TNF-alpha), Vascular cell adhesion molecule-1(VCAM-1), Lipid peroxide (malonaldehyde, MDA) in uncomplicated pregnancy. METHODS: Blood was collected from healthy women at 4 to 41 weeks' gestation and non-pregnant women. Plasma samples were measured by immunoassay for TNF-alpha, VACM-1 and by colorimetric assay for lipid peroxide, and data were statistically analyzed. RESULTS: Plasma concentration of TNF-alpha was not significantly elevated during first trimester compared with non-pregnant women, but significantly elevated during second and third trimester compared with non-pregnant women. Plasma concentration of VCAM-1 was significantly elevated during first trimester compared with non-pregnant women. Plasma concentration of lipid peroxide was not significantly elevated during pregnancy compared with non-pregnant women. CONCLUSION: The plasma concentration of TNF-alpha and VCAM-1 were significantly higher than that of non-pregnant state during second and third trimester in case of TNF-alpha, and during first trimester in case of VCAM-1. But the plasma concentration of lipid peroxide during pregnancy was not significantly different from that of non-pregnant, and the plasma concentration was kept up constant levels during gestation. These were seems to be meant that abnormal pregnancy would be happened if the level is above or below the measured level.
Cell Adhesion ; Female ; Humans ; Immunoassay ; Longitudinal Studies ; Necrosis ; Plasma* ; Pregnancy Trimester, First ; Pregnancy Trimester, Third ; Pregnancy* ; Tumor Necrosis Factor-alpha* ; Vascular Cell Adhesion Molecule-1*

OBJECTIVE: To investigate the relationship between the rotator cuff tear (RCT) and the muscle strength in hemiplegic side, and the effects of paralysis on the affected shoulders in hemiplegic patients. METHODS: A cross-sectional observational study performed in a university hospital was presented. The study enrolled 55 participants with hemiplegia of diverse degree of motor paresis, excluding those with bilateral lesions, history of major trauma or other co-existing musculoskeletal disorders of the shoulder. The main outcome measurements were muscle strength of the affected upper extremity (based on Medical Research Council scale), RCTs of the bilateral shoulders (by ultrasonography), and presence of shoulder pain, subluxation of the glenohumeral joint, passive range of motions, and subacromial spurs. RESULTS: Comparing each side of the shoulders, the prevalence of shoulder pain and supraspinatus muscle tear was higher (p<0.0001, p=0.007), and the range of motion was restricted (p<0.0001, p<0.0001, p<0.0001, p<0.0001) in the affected side. There was a significant trend toward higher prevalence of RCT and shoulder subluxation in the weaker shoulder (p=0.019, p<0.0001). In a multivariate analysis, Manual Muscle Test grade of less than three was an independent risk factor for RCT (p=0.025). CONCLUSION: RCT in hemiplegia had a linear trend with muscle weakness in the affected side and the degree of weakness was an independent risk factor for the occurrence of RCT. In addition, shoulder pain, limitation of range of motions, and RCT were more frequent on the hemiplegic side of the shoulders. It is the first study to reveal a linear trend between RCT and upper extremity weakness and will provide physicians an insight to the management of RCTs in hemiplegic patients.
Hemiplegia ; Humans ; Multivariate Analysis ; Muscle Strength ; Muscle Weakness ; Muscles ; Paralysis ; Paresis ; Prevalence ; Range of Motion, Articular ; Risk Factors ; Rotator Cuff ; Shoulder ; Shoulder Joint ; Shoulder Pain ; Tendon Injuries ; Upper Extremity

Klinefelter's syndrome (KFS) is a gonosomal aberration disease that occurs in males, and is characterized by 47, XXY karyotype, hypogonadism and a lack of secondary sexual characteristics. A potential link between this hormonally deficient syndrome and autoimmune disease, particularly systemic lupus erythematosus (SLE), has been reported. On the other hand, KFS is rarely reported to be accompanied by rheumatoid arthritis (RA), and there are no Korean cases reported. We report the first Korean case of a KFS patient with sero-positive RA and discuss the role of the pathogenesis of RA with KFS.
Aluminum Hydroxide ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Carbonates ; Hand ; Humans ; Hypogonadism ; Karyotype ; Klinefelter Syndrome ; Lupus Erythematosus, Systemic ; Male ; X Chromosome

BACKGROUND: The manifestation of Membranoproliperative glomerulonephritis (MPGN) is variable from asymptomatic hematuria, nephrotic syndrome to advanced chronic kidney disease. The etiology, pathogenesis, and treatment of MPGN are unclear. The incidence of idiopathic MPGN is also rare in Korea, and the clinical course was rarely studied. So we investigated clinical course of idiopathic MPGN type 1. METHODS: From March 1990 to November 2004, renal biopsy was done in about 1500 patients in Dong-A universty hospital. Among them, 16 patients were diagnosed as idiopathic MPGN. Fourteen patients were observed over 6 months. We analyzed clinical data of these patients retrospectively. RESULTS: Male to female ratio of total 16 patients is 2.2:1. Mean age of patients was 37.6 years (14-76 years) at the time of diagnosis. Thirteen cases (81%) had nephrotic range proteinuria, 8 cases (50%) had hypertention, 12 cases (75%) had edema, and 4 cases (25%) had decreased renal fuction (serum creatinine>1.2 mg/dL) at the time of diagnosis. Average observation time was 55.7+/-37.4 (6-122)months. Six cases ended up with progressive renal failure. Three cases achieved complete remission. Renal survival time for 50% of cases was 92 months. In our study, edema and decreased renal fucntion at the time of diagnosis were related with later development of progressive renal failure (p<0.05). Of the total 14 cases, 4 cases ware treated conservatively, 6 cases were treated with anti-platelet agents, and 4 cases were treated with anti-platelet agents and prednisone. In conservative treatment group, 1 case ended up with progressive renal failure and 1 case achieved complete remission. In anti-platelet agents treatment goup, 4 cases ended up progressive renal failure. In combined anti-platelet agents and prednisone treatment group, 1 case ended up progressive renal failure and 2 cases achieved complete remission. However, the effect of each treatment is inconclusive because the number of the patients are too small. CONCLUSIONS: We suspect that deceased renal function and edema at the time of diagnosis may be risk factor predicting progressive renal failure in patients with idiopathic MPGN type 1.
Biopsy ; Diagnosis ; Edema ; Female ; Glomerulonephritis ; Glomerulonephritis, Membranoproliferative* ; Hematuria ; Humans ; Incidence ; Korea ; Male ; Nephrotic Syndrome ; Prednisone ; Proteinuria ; Renal Insufficiency ; Renal Insufficiency, Chronic ; Retrospective Studies ; Risk Factors