BACKGROUND: DNA content analysis of human solid tumor is now widely performed by flow cytometric study. One of the most interesting and potentially observation in this field is that proliferative activity(S-Phase fraction of cell cycle) may profoundly affect the prognosis. METHOD: S-Phase fraction(SPF) have been measured by flow cytometric method using tumor cells isolated from paraffin embedded tissue. To evaluate the prognostic significance, SPF of small lung cancer cell was assessed in 42 patients who died after receiving anticancer chemotherapy. RESULTS: 1) Mean survival time of patients with small cell lung cancer was 190(± 156) days, Survival time were shortened, when TNM stage and PS scale were advanced. 2) Mean value of SPF of patients with small cell lung cancer was 27.4(±8.5)%. SPF had nothing to do with advance of TNM stage and PS scale. 3) In each identical TNM stage, there were not statistic significance between SPF and survival times. 4) There was a tendency like that higher SPF, better chemotherapeutic CONCLUSION: We could not find statistic significance between SPF and survival times, but SPF was a good predictive factor for chemotherapeutic response.
DNA ; Drug Therapy ; Humans ; Lung Neoplasms* ; Lung* ; Paraffin ; Prognosis ; Small Cell Lung Carcinoma ; Survival Rate

BACKGROUND: To study the prognosis of patients with lung cancer, many investigators have reported the methods to detect cell proliferation in tissues including PCNA, thymidine autoradiography, flow cytometry and Ki-67. PCNA, also known as cyclin, is a cell related nuclear protein with 36KD intranuclear polypeptide that is maximally elevated in S phase of proliferating cells. In this study, PCNA was identified by paraffin-embedding tissue using immunohistochemistry which has an advantage of simplicity and maintenance of tissue architecture. The variation of PCNA expression is known to be related with proliferating fraction, histologic type, anatomic(TNM) stage, degree of cell differentiation, S-phase fraction and survival rate. We analyzed the correlation between PCNA expression and S-phase fraction, survival. METHODS: To investigate expression of PCNA in primary lung cancer, we used immunohistochemical stain to paraffin-embedded sections of 57 resected primary non-small cell lung cancer specimen and the results were analyzed according to the cell type, cell differentiation, TNM stage, S-phase fraction and survival. RESULTS: PCNA expression was dMded into five group according to degree of staging(-, +, ++, +++,++++). Squamous cell type showed high positivity than in adenocarcinoma. Nonsignificant difference related to TNM stage was noticed. Nonsignificant difference related to degree of cell differentiation was noticed. S-phase fraction was increased wit advance of PCNA positivity, but t could not reach the statistic significance. The 2 year survival rate and median survival time were -50% 13 months, +75% 41.3 months, ++73% 33.6 months, +++67% 29.0 months, ++++25% 9 months with statistic significance (P<0.05, Kaplan-Meier, generalized Wilcox). CONCLUSION: From this study. PCNA expression was high positive n squamous cell cancer. And, there was no relationship between PCNA positivity and TNM stage, cellular differentiation or S-phase fraction. But, the patients with high positive PCNA staining showed poor survival rate than the patients with lower positive PCNA. It was concluded that PCNA immunostaining is a simple and useful method for survival prediction in paraffin embedded tissue of non-small cell lung cancer.
Adenocarcinoma ; Autoradiography ; Carcinoma, Non-Small-Cell Lung* ; Cell Differentiation ; Cell Proliferation ; Cyclins ; Flow Cytometry ; Humans ; Immunohistochemistry ; Lung Neoplasms ; Neoplasms, Squamous Cell ; Nuclear Proteins ; Paraffin ; Prognosis ; Proliferating Cell Nuclear Antigen* ; Research Personnel ; S Phase ; Survival Rate* ; Thymidine

Tricho-rhion-phalangeal syndrome is characterized by the triad of slow growing, brittle hair and early loss of hair, distinctive faces which include a long philtrum and pear-shape nose, and peripheral cone shape epiphysis with brachyphalangia. Tricho-rhion-phalangeal syndrome is probably not so much uncommon. The tricho-rhion-phalangeal syndrome, however, is not well recognized to orthopaedic surgeons due to the minor finger deformities. We report a case of tricho-rhion-phalangeal syndrome with brief review of literature.
Congenital Abnormalities ; Epiphyses ; Fingers ; Hair ; Lip ; Nose ; Surgeons

Arsenic trioxide (As2O3) was introduced into the treatment of refractory or relapsed acute promyelocytic leukemia and showed a striking effectiveness in China and United States multicenter study. However, the mechanistic basis for the carcinogenic or therapeutic effects of arsenics is still poorly understood. So, this study is performed to determine whether As2O3 induces apoptosis through intrinsic caspase cascades in acute promyelocytic leukemia HL-60 cells. MATERIALS AND METHODS: HL-60 cells were treated with As2O3 to investigate apoptosis through signaling of caspase cascades and mitochondrial dysfunction. RESULTS: As2O3 (>0.5 uM) decreased the viability of HL-60 cells in a dose-dependent manner, which was revealed as apoptosis shown chromatin condensation and ladder pattern DNA fragmentation. As2O3 increased the catalytic activity of caspase family cysteine proteases including caspase-3 and -9 proteases. Consistently, PARP, an intracellular biosubstrate of caspase-3 protease, was cleaved from 116 kDa to 85 kDa fragments. It also induced the change of mitochondrial membrane potential. Morever, As2O3 resulted in the increase of Bak. CONCLUSION: These data suggest that As2O3 induces apoptosis of HL-60 cells through activation of intrinsic caspase protease with mitochondrial dysfunction.
Apoptosis* ; Arsenic* ; Caspase 3 ; China ; Chromatin ; Cysteine Proteases ; DNA Fragmentation ; HL-60 Cells* ; Humans ; Leukemia, Promyelocytic, Acute ; Membrane Potential, Mitochondrial ; Peptide Hydrolases ; Strikes, Employee ; United States

No abstract available.
Animals ; Calcium ; Rats* ; Urinary Bladder*

No abstract available.
Apnea* ; Doxapram*

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

BACKGROUND: Interactions between nimodipine, a calcium channel blocker, used perioperatively for the treatment of subarachnoid hemorrhage, and vecuronium, rocuronium, and atracurium were studied with phrenic nerve-hemidiaphragm preparations of rats. METHODS: Male 200-300 g Sprague-Dawley rats were randomly allocated into four groups (control, NMD(5), NMD(50) and NMD(500) group, n = 10, respectively) according to the nimodipine concentration, and three groups (control, NMD(2D) and NMD(7D), n = 10, respectively) according to the pretreatment duration. A square wave 0.1 Hz supramaximal stimuli was applied to the phrenic nerve-hemidiaphragm preparation and the twitch height response was recorded with mechanomyography. The dose-response curves were measured, and ED(5), ED(50), ED(90), and ED(95) of each vecuronium, rocuronium, and atracurium in different concentrations of nimodipine of 5, 50, and 500 ng/ml and rocuronium in pretreatment with nimodipine 2.5 mg/kg/d for 2 and 7 days were calculated using an inhibitory sigmoid Emax model. RESULTS: The dose-response curves of rocuronium and atracurium were significantly shifted to the left in NMD(500) group, and significantly shifted to the right in NMD(7D) group (P < 0.05). In NMD(500) group, ED(50), ED(90), and ED(95) of rocuronium and atracurium were significantly reduced, and those of rocuronium in NMD7D group were significantly increased compared with the control group (P < 0.05). CONCLUSIONS: Nimodipine 500 ng/ml in the phrenic nerve-hemidiaphragm preparation of rat increased sensitivity to rocuronium and atracurium, and the dose-response curve was significantly shifted to the left, but following pretreatment for 7 days, nimodipine decreased the potency of rocuronium, and the dose-response curve was significantly shifted to the right.
Animals ; Atracurium* ; Calcium Channels ; Colon, Sigmoid ; Humans ; Male ; Nimodipine* ; Rats* ; Rats, Sprague-Dawley ; Subarachnoid Hemorrhage ; Vecuronium Bromide*

No abstract available.
Osteogenesis Imperfecta* ; Osteogenesis*