A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals.Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits,Ceditest(R)FMDV-NS (Ceditest(R) kit),UBI(R) FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI(R) kit) and a FMDV 3ABC-I-ELISA kitdeveloped at the Lanzhou Veterinary Research Institute.The test parameters (sensitivity and specificity) of the three kits were determined,and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits.The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest(R) kits was 98.05%,and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI(R) kits was 94.4%; the sensitivity of both Ceditest(R) and FMDV 3ABC-I-ELISA kit was 100%.However,the sensitivity of the UBI(R) kit was only 81.8%.With sera from naive or vaccinated non-infected animals,the specificity of all tests exceeded 90%.

OBJECTIVETo evaluate the safety of iodine-125 seed implantation in the liver.

METHODSTwenty New Zealand rabbits were divided into control and treatment groups and in the latter, iodine-125 seeds of 37 MBq were implanted into the liver under CT guidance whereas nonradioactive seeds were implanted in the control rabbits. Four weeks after implantation, white blood cell count, liver functions, and renal functions were measured or evaluated for comparison with those before implantation. The rabbits were then anesthetized to collect the liver tissue for pathological examination with HE staining and cell apoptosis assay.

RESULTSObvious hepatic tissue necrosis was observed around the radioactive seeds in the treatment group. At a 5 mm distance to the seeds, a distinct boundary occurred between the necrotic hepatic cells and normal cells. The control rabbits, however, had normal liver structure around the seeds implanted. In situ cell apoptosis examination showed a distinct band of apoptotic cells in the liver tissue of rabbits in the treatment group, which was not found in the control group. Two weeks after iodine-125 irradiation, alanine aminotransferase significantly increased in the treatment group (t=6.285, P<0.001), but recovered two weeks later (t=2.002, P=0.06). No significant alterations occurred in aspartate aminotransferase, blood urea nitrogen, serum creatinine, hemoglobin, serum total bilirubin, white blood cell count, or platelet count after the seed implantation.

CONCLUSIONIodine-125 seed implantation in the liver results in conformal irradiation dose distribution without obvious effects on the vital organs, demonstrating iodine-125 seed implantation as a safe and minimally invasive technique for hepatic cancer treatment.

Alanine Transaminase ; blood ; Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; In Situ Nick-End Labeling ; Iodine Radioisotopes ; adverse effects ; Liver ; pathology ; physiopathology ; radiation effects ; Male ; Rabbits ; Radiation Injuries, Experimental ; blood ; etiology ; pathology ; Random Allocation ; Time Factors

OBJECTIVETo investigate the changes of ataxia-telangiectasia mutated (ATM) phosphorylation in HepG(2) cells in relation to HepG(2) cell survival under continuous low dose-rate irradiation.

METHODSHepG(2) cells were exposed to equivalent irradiation doses delivered at either a continuous low dose-rate (7.76 cGy/h) or a high dose-rate (4500 cGy/h), and the phosphorylated ATM proteins and surviving fraction of HepG(2) cells after the exposures were compared.

RESULTSThe phosphorylation of ATM protein was maximal at 0.5 Gy irradiation delivered at either a high doserate or a continuous low doserate. As the radiation dose increased, ATM protein phosphorylation decreased under continuous low dose-rate irradiation, but remained stable under high dose-rate irradiation. With comparable ATM protein phosphorylation induced by continuous low dose-rate irradiation and high dose-rate irradiation, there was no significant difference in the surviving fraction of HepG(2) cells (P>0.05), but at a significantly lower ATM protein phosphorylation level than that induced by high dose-rate irradiation, continuous low dose-rate irradiation resulted in increased cell killing (P<0.01).

CONCLUSIONContinuous low dose-rate irradiation increases HepG(2) cells radiosensitivity as compared with high dose-rate irradiation. Increased cell killing following continuous low dose-rate irradiation is associated with reduced phosphorylated ATM protein, and inhibition of ATM phosphorylation may increase the radiosensitivity of HepG(2) cells.

Animals ; Ataxia Telangiectasia Mutated Proteins ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cell Survival ; radiation effects ; DNA-Binding Proteins ; metabolism ; Dose-Response Relationship, Radiation ; Humans ; Mice ; Phosphorylation ; radiation effects ; Protein-Serine-Threonine Kinases ; metabolism ; Radiation Tolerance ; radiation effects ; Time Factors ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo study the relationship between the degree of white matter damage and changes in brain function in premature infants early after birth according to amplitude-integrated electroencephalogram (aEEG) and raw EEG (with burst-suppression patterns).

METHODSThirty-eight premature infants of less than 32 weeks' gestational age and with white matter damage, including 20 cases of mild white matter damage and 18 cases of severe white matter damage, were included in the study. Forty-two premature infants without white matter damage were selected as a control group. After birth, they were examined using aEEG and brain ultrasound once a week until four weeks after birth or a corrected gestational age of 32 weeks. The white matter damage and control groups were compared in terms of aEEG patterns and amplitudes and burst suppression ratio (BSR) on EEG.

RESULTSThe white matter damage and control groups had highly discontinuous patterns and had no complete sleep cycles. The lower amplitude was significantly smaller in the severe white matter damage subgroup than in the mild white matter damage subgroup and control group. There was alternating burst-suppression activity on the raw EEG in the white matter damage and control groups; and the severe white matter damage subgroup had a significantly longer suppression time and a significantly higher BSR on EEG compared with the mild white matter damage subgroup and control group.

CONCLUSIONSBrain function monitoring should be performed in premature infants with white matter damage early after birth so as to detect cases of severe white matter damage in time.

Brain ; pathology ; Electroencephalography ; Humans ; Infant, Newborn ; Infant, Premature ; physiology ; Leukomalacia, Periventricular ; physiopathology

BACKGROUNDEndothelial progenitor cells (EPCs) derived from bone marrow may differentiate into endothelial cells and participate in endothelial repair. These cells can be mobilized into peripheral blood by cytokines, including granulocyte colony-stimulating factor (G-CSF). In the present study, we investigated the effects of G-CSF on neointimal formation and restenosis in a canine model of arterial balloon injury.

METHODSSixteen male beagle dogs were injected subcutaneously with 20 microg x kg(-1) x d(-1) recombinant human G-CSF (n = 8) or normal saline (n = 8) for 1 week. On the fifth day of treatment, the dogs underwent renal arterial angioplasty. At 8 weeks after arterial balloon injury, angiographic observations were made and injured arteries were processed for morphometric analysis of neointimal formation.

RESULTSPeripheral white blood cell counts were increased by 3.34-fold compared to baseline on the fifth day of administration of G-CSF. Angiographies revealed that one stenosis had occurred among the eight injured renal arteries from dogs treated with G-CSF, whereas all injured renal arteries from dogs treated with normal saline remained patent. The mean extent of stenosis among injured arteries was 18.3% +/- 17.9% in the G-CSF treated group compared to 12.5% +/- 7.6% in the saline treated control group (P = 0.10). G-CSF treatment slightly increased neointimal thickness (0.42 +/- 0.15 mm vs 0.25 +/- 0.06 mm, P = 0.08) with an intima to media ratio of 0.83 +/- 0.49 vs 0.54 +/- 0.18 (P = 0.11).

CONCLUSIONSG-CSF treatment does not attenuate neointimal hyperplasia and restenosis formation in a canine model of renal arterial injury, suggesting that the therapeutic strategy for preventing restenosis by stem cell mobilization should be investigated further.

Animals ; Dogs ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Hyperplasia ; Male ; Recombinant Proteins ; Renal Artery ; injuries ; pathology ; Tunica Intima ; pathology

Entire 3ABC sequence of FMDV containing a 6 x his tag coding sequence at the N-terminal was obtained through PCR amplification using a pair of specific primers, subcloned into shuttle plasmid of pMelBac-B with a melittin secretion signal sequence and finally constructed recombinant plasmid of pMel-3ABC. After co-transfected the recombinant plasmid and linearized Bac-N-Blue DNA into Sf9 insect cell under intermediary agent of the Cellfectin, the result showed that we have already acquired recombinant baculovirus by screen of plaque assay and identification of PCR. Though the recombinant baculovirus infecting the Sf9 cells again, experiments indicated that 3ABC gene could express in insect cells and the expressed protein was secreted in the supernatant of Sf9 cell culture possessing favourable biological activities detected by adopting two methods of SDS-PAGE and Western blot. The result verified that the protein could respond with sera derived from FMDV infected animals, but have no responsibility with sera derived from health animals and vaccinated animals detected by indirect ELISA using antigen of expressed protein after purification with Ni-NTA his bind resin. Therefore, this study has established a solid foundation for establishing an effective diagnosis method to discriminating the FMDV infected animals from vaccinated animals.
Animals ; Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Cattle ; Cell Line ; Cloning, Molecular ; Culture Media, Conditioned ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Foot-and-Mouth Disease ; immunology ; virology ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; metabolism ; Gene Expression Regulation, Viral ; Immune Sera ; immunology ; Plasmids ; genetics ; Recombinant Proteins ; immunology ; secretion ; Sheep ; Spodoptera ; Swine ; Transfection ; methods ; Viral Nonstructural Proteins ; genetics ; immunology ; secretion

To investigate the influence of G-CSF mobilization on functions of donor T lymphocyte subpopulation and acute graft-versus-host disease, peripheral blood samples of 20 healthy donors were collected before and after G-CSF mobilization. The whole blood was diluted with IMDM in ratio of 1:1 and then incubated with PMA + ionomycin + monensin at 37 degrees C, 5% CO2 for 4 hours. After being mobilized and stained, the IL-4, IFN-gamma and IL-2 positive cells were counted with three-color flow cytometry. The results showed that before G-CSF mobilization, the percentages of donor's CD3(+)IFN-gamma(+), CD4(+)IFN-gamma(+), CD8(+)IFN-gamma(+) T cells were 3.2% (0% - 45.9%), 1.3% (0% - 23.8%) and 1.5% (0% - 22.2%) respectively. The percentage of above mentioned cells in donor increased to 19.2% (0% - 53.9%), 9.5% (0% - 49.5%), 7.5% (0% - 38.1%) respectively after G-CSF mobilization. The IL-2 positive CD3(+), CD4(+) and CD8(+) T cell percentage in pre-G-CSF mobilized donors was 1.5% (0% - 31%), 0.8% (0% - 30.0%) and 0% (0% - 5.3%) respectively and subsequently increased to 25.7% (0% - 51%), 19.8% (0% - 39.7%), 4.6% (0% - 20.9%) respectively after G-CSF mobilization. The IL-4 positive T subpopulation did not increased significantly after G-CSF mobilization. In the early stage after peripheral blood stem cell transplantation, donor's Tc1 percentage in aGVHD group was significantly higher than that in non-aGVHD group. The morbidity of severe aGVHD in high Tc2 percentage group was significantly lower than that in low Tc2 percentage group. It is concluded that the donor's type I T cells increase after G-CSF mobilization, the Tc1 percentage of G-CSF mobilized donor is correlated with the occurrence of aGVHD in the early stage after HSCT, the percentage of Tc2 in donor is negatively correlated with aGVHD morbidity in recipients.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; adverse effects ; Hematopoietic Stem Cell Mobilization ; adverse effects ; methods ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology

China ; Manganese/toxicity* ; Stainless Steel

Female ; Glomerulonephritis, IGA ; metabolism ; pathology ; Glomerulonephritis, Membranous ; metabolism ; pathology ; Glomerulosclerosis, Focal Segmental ; metabolism ; pathology ; Humans ; Kidney ; metabolism ; pathology ; Male ; Renal Insufficiency, Chronic ; metabolism ; pathology