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

Gastric web is a rare gastric anomaly and almost all the reported cases have been located in the distal antrum. We experienced a case of neonatal gastric web that was located in the body of stomach, and we report the findings of the upper gastrointestinal series and ultrasonography.
Stomach* ; Ultrasonography

Gastric volvulus is a rare condition, and it is classified as the organoaxial or mesentericaxial type according to the axis of rotation. We experienced 1 case of pediatric recurrent mesenteroaxial gastric volvulus and we report here the ultrasonographic and CT findings.
Axis, Cervical Vertebra ; Stomach Volvulus*

Multiseptate gallbladder is a very rare congenital malformation, and has never been reported in a neonate. We present the first published ultrasonographic findings of multiseptate gallbladder in a neonate.
Gallbladder* ; Humans ; Infant, Newborn*

Backgound: The placement of stents in coronary arteries has been shown to reduce acute closure and restenosis in comparison to balloon angioplasty. However, clinical use of intracoronary stents is impeded by the subacute stent thrombosis and hemorrhagic complications associated with the anticoagulant regimen. It's known that the complete stent deployment with high pressure inflation and new antiplatelet agents are effective in reduction of subacute thrombosis and hemorrhage. So we evaluated initial results (success and complication rate) after high pressure-stent deployment with new anticoagulation protocol. METHODS: One hundred and ninety one patients with 201 lesions were treated with 231 stents of various types. The high pressure balloon inflation and antiplatelets agents were used in all cases. Final high pressure balloon inflation guided by IVUS were performed in 23 consecutive cases with incomplete stent deployment according to angiographic findings. RESULTS: 1) The indications of stenting (n=210) were De novo in 124 (59%), bailout procedure in 57 (27%), suboptimal result after PTCA in 19 (8%), and restenosis after PTCA in 14 (6%). The location of lesions were LAD in 101, RCA in 67, circumflex in 28, ramus intermedius in 3, and LMT artery in 2 lesions. Angiographic morphologic characteristics were type A in 2, type B in 158 (B1: 57, B2: 101), and type C in 22 lesions. 2) The angiographic and clinical success rate was 96% (192/201) and 92% (186/201) respectively. 3) In angiographic analysis, the baseline average reference vessel dirmeter was 3.33+/-0.35 mm. Baseline minimum lumen diameter (MLD) was 0.58+/-0.29 mm, with baseline percent diameter stenosis of 82.86+/-8.64%. The final stent diameter was 3.37+/-0.29 mm, with mean final percent stenosis of 0.63+/-8.25. The mean MLD after stenting was significantly increased (p<0.001). The mean MLD within stent increased 14%, from 2.91+/-0.39 mm at the nominal balloon inflation (inflation pressure=7 atm) to 3.37+/-0.29 mm at high pressure balloon inflation (inflation pressure <0A65B>12atm) (p<0.001). The length of lesions in GR I (cook), GR II, and Micro II stents were significantly longer than ones in PS, Cordis, Wiktor, Nir (p<0.001). 4) In intravascular ultrasound analysis, the mean lumen CSA at the tightest point within stent increased 11%, from 8.4+/-2.4 mm2 at the intial intravascular ultrasound to 9.4+/-2.1 mm2 at the final intravascular ultrasound (p<0.001). 5) The procedural and postprocedural complications were 2 acute closures associated with AMI and emergent CABG, 1 subacute closure which was revascularized by bail out stenting, 5 major hemorrhage requiring transfusion associated with 1 CVA and 2 metabolic acidosis induced by acute renal failure, and 5 death. CONCLUSION: The high pressure stent deployment procedure and new anticoagulation protocol associating tidopidine and aspirin without coumadin or prolonged heparin infusion allow us to obtain an acceptably low subacute thrombosis or bleeding complication rate. These results are encouraging and allow a wide use of coronary stenting.
Acidosis ; Acute Kidney Injury ; Angioplasty, Balloon ; Arteries ; Aspirin ; Constriction, Pathologic ; Coronary Vessels* ; Hemorrhage ; Heparin ; Humans ; Inflation, Economic ; Platelet Aggregation Inhibitors* ; Stents* ; Thrombosis ; Ultrasonography ; Warfarin

During resection of the femoral artery aneurysm under Halothane-N2O-O2 anesthesia, the authors experienced accidental autotransfusion in which the autologous blood aspirated and collected in plastic bag, were transfused to the patient without hematologic test. But there were no clinical problems without mild bleeding tendency.
Anesthesia ; Aneurysm ; Blood Transfusion, Autologous* ; Femoral Artery ; Hematologic Tests ; Hemorrhage ; Humans ; Plastics

No abstract available.
Facial Nerve* ; Paralysis* ; Transplants*

Kaposi's sarcoma was first descrided by Kaposi in 1872 as an idiopathic multiple hemorrhagic sarcoma. Its clinical features revealed to be erythematous red or purple macule started out, and developing into palpable dome-shaped nodules. Etiology is not defined to detail at present. Kaposi's sarcoma is classified to 4 categories; Classical, African, Epidemic and Transplant type. Epidemic or AIDS categories is found approximately 20 % of all AIDS and has strong predilection for head and neck region. The first case of Kaposi's sarcoma observed in patients with kidney transplants was reported in 1969. Kaposi's sarcoma now accounts for 5% of all tumors associated with transplanted patients and alteration of the immunosuppression may have played a key role in these recipients. The most common site of Kaposi's sarcoma in transplanted patients are extremities but rare in head and neck area. We report a case of Kaposi's sarcoma on the hard palate in the kidney transplantation patient.
Extremities ; Head ; Humans ; Immunosuppression ; Kidney ; Kidney Transplantation ; Mouth* ; Neck ; Palate, Hard ; Sarcoma ; Sarcoma, Kaposi*

No abstract available.
Multiple Myeloma* ; Plasmacytoma*