No abstract available.

No abstract available.

In this study, we are to produce the single chain variable fragment (scFv) antibodies against surface protein of hepatitis B virus (HBV) using antibody phage display technique. Balb/c mice were immunized with preS1 and cDNAs of heavy and light chains of splenic B cells from immunized mice were prepared using RT-PCR. Two cDNAs were linked with (64S) linker DNA under recombination PCR to produce single chain Fv DNA. After digestion of scFv DNA with Sp 1 and Not 1, the digested DNA was ligated into pCANTAB 5E and electroporated into E. coli XL1-Blue to prepare scFv-library. The size of library was 1 * 10' pfu/ml. Phage antibodies (phabs) against preS1 were rescued with M13K07 helper phages, and preS1-binders were selected through 3 times of panning using 96 well microtitre plates. Phage antibody clones were assayed directly for the ability to bind preS1 by ELISA. And then 7 phage antibody clones had high ELISA signals against preS1. Phabs from preS1-specific pMsc-17 had the strongest ELISA signal to preS1. Phabs from pMsc-17 were used for Western blot to preS1 and the results revealed that it was specific to preS1. To prepare the soluble scFv antibody, phabs from pMsc-17 were transfected into non-suppressor E. coli HB2151, and grown under 1 mM IPTG. Soluble scFv antibody was mainly accumulated in the periplasmic space, but small amount of antibody was secreted into culture media.
Animals ; Antibodies ; B-Lymphocytes ; Bacteriophages* ; Blotting, Western ; Cell Surface Display Techniques ; Clone Cells ; Culture Media ; Digestion ; DNA ; DNA, Complementary ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Isopropyl Thiogalactoside ; Mice* ; Periplasm ; Polymerase Chain Reaction ; Recombination, Genetic ; Single-Chain Antibodies*

Human monoclonal antibodies have considerable potential in the prophylaxis and treatment of viral disease. By cloning human Ig gene segments from the B cells of volunteer into pComb3 phagemid vector, antibody library was created of filamentous phage particles displaying Fab fragments on their surface after being rescued with M13KO7 helper phages. The size of library was 7x10' pfu. Phage antibodies (phabs) were panned against biotinylated preS1 using streptavidine coated Dynabead. The soluble Fab antibodies were prepared from phagemid colonies and assayed directly for the ability to bind preS1 by ELISA. And then 3DW and SGW specific to preS1 which have both heavy and light chain to form Fab fragment, were selected. The soluble Fab antibody from 3DW was expressed highly at the concentration of 0.1 - 1.0 mM of IPTG, and 5 hours postinduction. The soluble antibodies from 3DW and SGW showed their relative affinities of 2x10' M ', and Sx10 M ', respectively, and the specificities to preS1 on ELISA. Our results suggest that antibody phage display library is very useful method to generate the human monoclonal antibody and that the human Fab monoclonal antibodies specific to preS1 selected in this study open the way to treat hepatitis B as a component of passive irnmunotherapeutics.
Antibodies ; Antibodies, Monoclonal ; B-Lymphocytes ; Bacteriophages* ; Clone Cells ; Cloning, Organism ; Enzyme-Linked Immunosorbent Assay ; Genes, Immunoglobulin ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Humans* ; Immunoglobulin Fab Fragments ; Isopropyl Thiogalactoside ; Streptavidin ; Virus Diseases ; Volunteers

A 25-year-old 7 month pregnant woman with mitral stenosis was hospitalized because of life threatening hemoptysis dispite intensive medical treatment. Emergency mitral commissurotomy resulted in prompt cessation of hemorrhage and at nine months follows-up, the hemoptysis has not recurred. Mitral valve surgery appears to be consitently associated with rapid and sustained cessation of hemoptysis. Therefore, surgery should be considered in hemoptysis due to mitral stenosis, particulary when the danger of asphyxiation exists.
Adult ; Emergencies* ; Female ; Hemoptysis* ; Hemorrhage ; Humans ; Mitral Valve ; Mitral Valve Stenosis* ; Pregnant Women

To clarify possible causes, pathogenesis, and appropriate prevention method of remote intracerebral hemorrhage (RICH), we analyzed the clinical findings and the possible predisposing factors of six cases who developed RICH among the 206 surgical series of cerebral aneurysm operated in our hospital over recent 5-year period. The locations of aneurysm were anterior communicating artery in three cases and internal carotid artery in three other cases. The sites of RICH were dependent regions considering the operative position in five of six cases, con-tralateral cerebellum in three cases, bilateral cerebellum in one case, and contralateral occipital area in one case. Peripheral low density around the RICH, suggesting hemorrhagic infarction, was observed on computed tomography in four cases. No patient had preoperative hypertension; however, significant elevation of blood pressure was observed intraoperatively or postoperatively in three cases. The factors which could have induced brain shift(large amount of removed or drained cerebrospinal fluid, large amount of infused mannitol, too low PaCO2) were observed in all cases except one case. There was no case with coagulopathy or underlying occult lesion. Two patients in whom detection of RICH was delayed showed poor outcomes. The possible underlying mechanisms involved in such complication seem to be shifting of brain due to sudden decreased intracranial pressure and excessive removal of cerebrospinal fluid, and subsequent injury of blood vessels such as compression or breakdown of vein. Moreover, the sudden elevation of blood pressure may have played a role as contributing factor. Therefore, consideration should be given to the maintenance of an adequate volume of intracranial cerebrospinal fluid and the appropriate blood pressure to prevent this complication. Early detection and immediate treatment with awareness of the possibility of this complication should be borne in mind when treating these patients to prevent such complications and to obtain good results.
Aneurysm* ; Arteries ; Blood Pressure ; Blood Vessels ; Brain ; Carotid Artery, Internal ; Causality ; Cerebellum ; Cerebral Hemorrhage* ; Cerebrospinal Fluid ; Hemorrhage ; Humans ; Hypertension ; Infarction ; Intracranial Aneurysm ; Intracranial Pressure ; Mannitol ; Veins

Since introduction of magnetic resonance imaging(MRI), six cases of multiple intracranial meningiomas without signs of neurofibromatosis were found among the forty eight cases of meningiomas. The incidence of multiple intracranial meningiomas in our series was 12.5% of all meningiomas. We analyzed six cases of multiple intracranial meningiomas that were evaluated by MRI. Two tumors were found in five patients, whereas the other one patient had more than ten tumors at different sites. The tendency of unihemispheric distribution was not seen and two cases had bilaterally located tumors and other two cases had tumors above and below the tentorium without tentorial involvement. Relatively high occurrence of meningioma in posterior fossa was observed. Half of the six cases had posterior fossa tumors and 6 tumors(28.6%) of the total 21 tumors were located in the posterior fossa. All the cases were female. Mean age on admission was 61.3 years, ranged from 44 to 77 years. All six patients were treated surgically and three of them underwent multiple staged operations. Histologically, of the 12 tumors removed, 5 were transitional, 4 were fibroblastic, 3 were meningothelial. The histological subtypes of tumors in each case were different in two cases, but identical in other three cases. The surgical results, although the follow-up periods(ranged 3 months to 2.5 years) were not long enough in some cases, were favorable.
Female ; Fibroblasts ; Follow-Up Studies ; Humans ; Incidence ; Infratentorial Neoplasms ; Magnetic Resonance Imaging ; Meningioma* ; Neurofibromatoses

No abstract available.
Humans ; Risk Factors*

In order to observe the influence of halothane upon the blood pressure during general anesthesia, the author prepared a formula, expressing change of blood pressure by times series and analyzed the types and distribution patterns in this experiment. The results obtained were as follows: 1) Blood pressure was high at the time of intubation. 2) Generally, blood pressure increased at the time of intubation and then stabilized within 20 minutes. 3) Most common patterns were dentified. (ijEE) type was 73~74% that is most common type, and (iEEE) type was 40~44%.
Anesthesia* ; Anesthesia, General ; Blood Pressure* ; Halothane* ; Intubation

No abstract available.