Neomorphogenesis of cartilage using chondrocyte-polymer constructs is a potential source for development of cartilage reconstruction. Current tissue engineering techniques of neocartilage rely on in vivo implantation of polymer-chondrocyte constructs. The purpose of this study was to find a way to bioengineer cartilage in vitro by entrapping chondrocytes in a molded autologous fibrin glue. Chondrocytes isolated from the cartilage of rabbit joints were combined with fibrinogen extracted by a single cryoprecipitation of autologous plasma, and they were then polymerized with thrombin to create a fibrin glue with a final cell density of 2.5x10(6) cells/ml. The collagen for a control study was used as a polymer. The polymer-chondrocyte constructs were cultured for 4 weeks and the fibrin-chondrocyte constructs molded in the shape of a human ear were cultured for 6 weeks in vitro. Morphometric, histochemical, and histomorphometric analysis including glycosaminoglycan quantitation confirmed the following results: 1) Highly-concentrated autologous fibrinogen was easily extracted by a single cryoprecipition of autologous olasma. 2) The fibrin-chondrocyte constructs demonstrated the presence of actively proliferating chondrocytes with the production of cartilaginous matrix(collagen and glycosaminoglycan) at 1 week after culture, as well as gross and histologic evidence similar to those of normal cartilage at 3-4 weeks after culture. 3) The collagen-chondrocyte constructs demonstrated lower degrees of hardness and transparency, as well as a lower density of cells and glycosaminoglycan during the culture period. 4) Neocartilage generated from fibrin-chondrocyte constructs in the shape of a human ear nearly retained their original configuration and size without degeneration for 6 weeks of culture in vitro. This study demonstrated a novel method for bioengineering the molded cartilage in vitro using autologous fibrin glue as a matrix scaffold. The generated cartilage showed gross and histologic evidence similar to those of normal cartilage, retaining the original gross dimension. With further refinement, this may be a new application of tissue engineering for the reconstruction of cartilage.
Bioengineering ; Cartilage* ; Cell Count ; Chondrocytes* ; Collagen ; Ear ; Fibrin Tissue Adhesive* ; Fibrin* ; Fibrinogen ; Fungi ; Hardness ; Humans ; Joints ; Plasma ; Polymers ; Thrombin ; Tissue Engineering*

The ambulatory electrocardiographic examinations were performed in 31 patients (mean age of 59.4+/-9.3 yrs : male 16 cases, female 15 cases) with ischemic heart disease to evaluate the clinical features of ST segment more than 1 mm persisting for 45 seconds or longer. The incidence of associated disease are angina pectoris 14 cases, acute myocardial infarction 3 cases, old myocardial infarction 7 cases, hypertension 19 cases, diabetes mellitus 5 cases, cerebrovascular disease 4 cases, aortic regurgitation 2 cases, ventricular arrhythmia 1 case and chronic renal faliure 1 case. 93.7% of 252 monitored episodes of transient myocardial ischemia were silent. The incidence and duration of transient myocardial ischemia were 8.1+/-6.7 episodes/day (7.6+/-6.5episodes/day for silent myocardial ischemia, 0.5+/-0.9 episodes/day for silent ischemia, 7.6+/-14.1mins/day for symptomatic ischemia). The heart rate at the onset of ST segment depression is higher in symptomatic episode than silent episode (94.6+/-19.7 vs 82.1+/-17.4/min,. p<0.05). But duration of ST segment depression is longer in silent episode than symptomatic episode(32.4+/-97.7 vs 14.8+/-10.2/min,. p<0.01). Maximal ST segment depression was similar between silent and symptomatic episode (1.61+/-0.65 mm, 1.97+/-0.84 mm, repectively). 55.5% of silent episodes occurred during sleep or resting state and 60% of symptomatic episodes occurred during strenuous effort, exercise or eating (p<0.01). Transient myocardial ischemia developed not more frequently in the morning probably because the 24 hour Holter electrocadiographic examination was performed during hospitalization in the majority of cases.
Angina Pectoris ; Aortic Valve Insufficiency ; Arrhythmias, Cardiac ; Depression ; Diabetes Mellitus ; Eating ; Electrocardiography ; Female ; Heart Rate ; Hospitalization ; Humans ; Hypertension ; Incidence ; Ischemia ; Male ; Myocardial Infarction ; Myocardial Ischemia*

Retroperitoneal duodenal rupture is rare and is often difficult to diagnose on the plain abdominal x-ray. From a review of the plain abdomen films of 21 cases with retroperitoneal duodenal rupture, confirmed by operation, pneumoretroperitoneum was revealed in 16 cases; Air in the peritoneum was manifested as a bubbly shadow in 12 cases, a renal halo in 9 cases, air shadow along the right psoas margin in 2 cases, air along the diaphragmatic crus in 2 cases and air in the right properitoneal fat in 2 cases, US and CT also revealed air bubbles and fluid collection around the right kidney. We recommend the plain abdomen as a useful diagnostic method for detection of pneumoretroperitoneum.
Abdomen* ; Kidney ; Methods ; Peritoneum ; Retropneumoperitoneum ; Rupture*

Mannitol is an osmotic diuretic agent useful in a variety of clinical conditions. This study is based on acid-base and electrolyte changes seen after the intravenous infusion of hypertonic mannitol for the prevention of cerebral edema. The study subjects were divided into 3 groups: for group A, an amount of 300-900 mL 15% mannitol was intravenously infused over the period of 60 to 90 minutes; for group B, 1,200-2,600 mL over 12 to 24 hours; and for group C, 3,200-4,900 mL over more than 24 hours. In group A, blood pH is increased from 7.43+/-0.07 to 7.46+/-0.04, and plasma HCO3- from 25.3+/-2.1 to 28.9+/-2.9 mEq/L, but plasma K+ is decreased from 4.3+/-0.6 to 3.7+/-0.8 mEq/L. In group B, blood pH is increased from 7.42+/-0.02 to 7.47+/-0.06, and plasma HCO3- from 25.2+/-1.8 to 29.1+/-2.9 mEq/L, but plasma K+ is decreased from 4.2+/-0.3 to 3.8+/-0.5 mEq/L. In group C, blood pH is increased from 7.41+/-0.01 to 7.52+/-0.04, and plasma HCO3- from 24.9+/-1.2 to 27.7+/-2.5 mEq/L, but plasma K+ is decreased from 4.2+/-0.1 to 3.9+/-0.2 mEq/L. These results showed that intravenous infusion of mannitol could induce metabolic alkalosis and hypokalemia, regardless of its dose. The mannitol induced metabolic alkalosis may be due to increased renal HCO3- production.
Alkalosis* ; Brain Edema ; Hydrogen-Ion Concentration ; Hypokalemia ; Infusions, Intravenous ; Mannitol ; Plasma

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*

No abstract available.
Aged ; Anti-Bacterial Agents/*adverse effects ; Anticonvulsants/therapeutic use ; Cephalosporins/*adverse effects ; Consciousness Disorders/diagnosis/drug therapy/*etiology ; Diabetic Nephropathies/complications/*therapy ; Electroencephalography ; Female ; Humans ; Pneumonia, Bacterial/complications/*drug therapy ; *Renal Dialysis ; Status Epilepticus/diagnosis/drug therapy/*etiology ; Treatment Outcome ; Uremia/therapy

All urate transport occurs across the renal epithelial cells of the proximal tubule. Most of the filtered urate is reabsorbed in the S1 segment of the early proximal tubule. This is followed by tubular secretion in the S2 segment of the proximal tubule and approximately 50% of the filtered urate flows back into the tubular lumen. Most of the secreted urate undergoes postsecretory reabsorption that occurs predominantly in the last S3 segment of the proximal tubule. Recently, four proteins that transport urate have been identified at the molecular level. These proteins are an electrogenic urate uniporter, urate transporter/channel (UAT), two members of the organic anion transporter (OAT) family, OAT1 and OAT3, and a protein with some homology to OAT4, designated URAT1.
Epithelial Cells ; Humans ; Ion Transport ; Uric Acid*

PURPOSE: To investigate whether growth hormone (GH) has a protective effect on neurons in hippocampal slice cultures of neonatal rats exposed to oxygen-glucose deprivation (OGD). METHODS: Cultured hippocampal slices of 7-day-old rats were exposed to OGD for 60 min. Then, the slices were immediately treated with three doses of GH (5, 50, or 500 micrometer) in media. The relative fluorescent densities of propidium iodide (PI) uptake in the slices and relative lactate dehydrogenase (LDH) activities in the media were determined and compared between each GH-treated group of slices and untreated slices (control) at 12 and 24 h after OGD. Immunofluorescent staining for caspase-3 and TUNEL staining were performed to observe the effect of GH on apoptotic neuronal death. RESULTS: The relative fluorescent densities of PI uptake in CA1 and dentate gyrus (DG) of the hippocampal slices in each GH-treated group were not significantly different from those in the untreated slices at 12 and 24 h after OGD (P>0.05). Treatment with GH could reduce the relative LDH activities in the media of the GH-treated groups only at 12 h after OGD (P<0.05). Expression of caspase-3 and TUNEL positivity in CA1 and DG of the slices treated with 50-iM GH were not different from those of the untreated slices at 12 and 24 h after OGD. CONCLUSION: Treatment of hippocampal slice cultures with GH after OGD does not show a definitive protective effect on neuronal death but can reduce the LDH efflux of the slices in media at 12 h after OGD.
Animals ; Apoptosis ; Caspase 3 ; Dentate Gyrus ; Growth Hormone ; In Situ Nick-End Labeling ; L-Lactate Dehydrogenase ; Neurons ; Propidium ; Rats

PURPOSE: To investigate whether growth hormone (GH) has a protective effect on neurons in hippocampal slice cultures of neonatal rats exposed to oxygen-glucose deprivation (OGD). METHODS: Cultured hippocampal slices of 7-day-old rats were exposed to OGD for 60 min. Then, the slices were immediately treated with three doses of GH (5, 50, or 500 micrometer) in media. The relative fluorescent densities of propidium iodide (PI) uptake in the slices and relative lactate dehydrogenase (LDH) activities in the media were determined and compared between each GH-treated group of slices and untreated slices (control) at 12 and 24 h after OGD. Immunofluorescent staining for caspase-3 and TUNEL staining were performed to observe the effect of GH on apoptotic neuronal death. RESULTS: The relative fluorescent densities of PI uptake in CA1 and dentate gyrus (DG) of the hippocampal slices in each GH-treated group were not significantly different from those in the untreated slices at 12 and 24 h after OGD (P>0.05). Treatment with GH could reduce the relative LDH activities in the media of the GH-treated groups only at 12 h after OGD (P<0.05). Expression of caspase-3 and TUNEL positivity in CA1 and DG of the slices treated with 50-iM GH were not different from those of the untreated slices at 12 and 24 h after OGD. CONCLUSION: Treatment of hippocampal slice cultures with GH after OGD does not show a definitive protective effect on neuronal death but can reduce the LDH efflux of the slices in media at 12 h after OGD.
Animals ; Apoptosis ; Caspase 3 ; Dentate Gyrus ; Growth Hormone ; In Situ Nick-End Labeling ; L-Lactate Dehydrogenase ; Neurons ; Propidium ; Rats

PURPOSE: We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. METHODS: The hippocampus of 7-day-old rats was cut into 350 micrometer slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into 15 micrometer thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. RESULTS: 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P< 0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. CONCLUSION: The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.
Animals ; Anoxia ; Apoptosis ; Brain ; Caspase 3 ; Cell Death ; Fluoresceins ; Fluorescence ; Fluorescent Antibody Technique ; Hippocampus ; In Situ Nick-End Labeling ; Ischemia ; Neurons ; Propidium ; Pyramidal Cells ; Rats ; Reperfusion