OBJECTIVETo elucidate the evolution pattern of human influenza virus A H3 subtype by detecting positive selected codons in hemagglutinin gene.

METHODSAll H3 sequences in NCBI GenBank and influenza sequence database were downloaded and two step cluster method was applied to divide sequences into six groups, which were corresponding to different period by turns. Fixed Effect Model was applied to detect positive selected codons in each group, and two step cluster method was then used again to summarize variation patterns of selective pressure among sites.

RESULTSPositive selected codons were different in groups corresponding different periods. 50 amino acid codons had been identified as positive selected sites in at least one time span. Among them, 42 codons belonged to one of the five known antigen-combinng regions. A larger amount of sites as well as relatively higher selection pressure were identified in antibody combining regions A and B. Results showed that the 50 sites could be divided into seven different patterns. While other six patterns corresponding to positive selected codons at only one time span, the sites of the seventh pattern were under positive selection in several periods.

CONCLUSIONPositive selection codons in evolution of H3A1 strains were alternated in different time period whereas antibody combining regions A and B played more important roles in the evolution process. Other 8 identified codons out of the antibody combining regions might belong to unknown antigen regions.

Amino Acid Sequence ; Codon ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; Influenza, Human ; genetics ; Selection, Genetic

OBJECTIVETo investigate the expression changes of intrinsic cytokines TGF-beta(1) and TNF-alpha, telomerase activity and bcl-2 during ongoing apoptosis of HL-60 and K562 cells induced by p53.

METHODSpN53cG (Val135), a temperature sensitive p53 mutant, which behaved like wild type p53 (wt-p53) at 32.5 degrees C, were introduced into p53-null HL-60 and K562 cells respectively by lipofectin. In the presence of G418, HL-60-pN53cG and K562 pN53cG clones expressing p53 protein were selected. The ongoing expression of intrinsic cytokines (TGF-beta(1) and TNF-alpha), bcl-2 oncogene and hTERT mRNA during the apoptosis of HL-60 and K562 cells induced by p53 and the effects of exogenous p53 gene, TGF-beta(1) and TNF-alpha antisense PS-ODNS on the apoptosis of HL-60 and K562 cells and the expression of bcl-2 were studied by RT-PCR, quantitative RT-PCR, DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometery. The levels of secreted TGF-beta(1) and telomerase activity were detected by ELISA and PCR-ELISA, respectively.

RESULTS(1) The expressions of intrinsic TGF-beta(1) and TNF-alpha mRNA were up-regulated, while that of bcl-2 and hTERT down-regulated. The levels of TGF-beta(1) in the supernatant of HL-60 and K562 cells were increased, and the level of telomerase activity decreased. (2) Antisense PS-ODNS of TGF-beta(1) and TNF-alpha could obviously inhibit the p53 inducing cell apoptosis, and restore bcl-2 mRNA and protein to pre-treated level.

CONCLUSIONSExogenous p53 induces leukemia cell apoptosis via up-regulating the expression of intrinsic TGF-beta(1) and TNF-alpha and down-regulating the expression of hTERT and bcl-2.

Apoptosis ; drug effects ; genetics ; DNA, Antisense ; pharmacology ; DNA-Binding Proteins ; Gene Expression Regulation, Neoplastic ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Mutation ; Plasmids ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection ; methods ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; genetics ; Tumor Suppressor Protein p53 ; genetics ; physiology

OBJECTIVETo study the possible relationship between coronary artery lesions and fibrinogen Bbeta-148 C/T polymorphism in children with Kawasaki disease.

METHODSFast blood samples were taken from 36 children with Kawasaki disease (21 had coronary artery lesions) and 49 age- and gender-matched healthy children (control group). Plasma levels and molecular reactivity of fibrinogen were measured with Assist Plasma Fibrinogen Activity Assay System. Polymerize chain reaction and restriction enzyme digestion were used to detect the genotypes of fibrinogen Bbeta-148C/T gene polymorphism.

RESULTSThe plasma fibrinogen levels in patients with coronary artery lesions were significantly higher than those in patients without coronary artery lesions and in the control group. T allele frequency in patients with Kawasaki disease was significantly higher than that in the control group. The patients with coronary artery lesions had more increased T allele frequency compared with the patients without coronary artery lesions.

CONCLUSIONSPlasma fibrinogen levels and fibrinogen Bbeta-148C/T polymorphism are associated with coronary artery lesions in children with Kawasaki disease.

Child, Preschool ; Coronary Artery Disease ; etiology ; Female ; Fibrinogen ; analysis ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; genetics ; Polymorphism, Genetic

Long-term potentiation (LTP) is thought as a generative mechanism underlying learning and memory via storing information in central nervous system. Electro-neurophysiological assay for LTP is generally used in screening the drugs that can facilitate learning and memory. By using in vivo LTP technique, isolichenin was found to facilitate LTP induction by a tetanic stimulation (20 pulses/100 Hz) in dentate gyrus. This tetanic stimulation by itself, however, cannot induce LTP. Previous study showed the reagent being able to facilitate LTP-induction, like methanol extract of saffron (MES), usually can antagonize the inhibiting effect of 30% ethanol on LTP induction (30 pulses/60 Hz). Isolichenin may also fall into such kind of drugs. Interestingly, comparatively study showed that isolichenin failed to antagonize the inhibiting effect of 30% ethanol on LTP induction (30 pulses/60 Hz). This result indicates a different unknown mechanism existing in the effect of isolichenin on LTP or memory formation.
Animals ; Crocus ; chemistry ; Dentate Gyrus ; drug effects ; physiology ; Long-Term Potentiation ; drug effects ; physiology ; Male ; Plant Extracts ; pharmacology ; Polysaccharides ; pharmacology ; Rats ; Rats, Wistar

BACKGROUNDTo find out the timing of serologic responses after illness onset and distribution of IgG antibody to SARS-CoV in SARS cases of transmission chain or non-transmission chain.

METHODSThe IgG and IgM antibodies to SARS-CoV were tested by indirect ELISA in serum samples from 301 clinically diagnosed SARS cases.

RESULTSTotally 158 SARS cases were involved in 15 chains of transmission. The positive rates of SARS-CoV IgG in those chains were 85.70%-100.00% and the overall rate was 94.30% (149/158). The chain of transmission could spread to four generations, but the SARS cases were reduced with increase of generations. There was no significant difference among positive rates of SARS-CoV IgG for generations, Chi square=5.11, P greater than 0.05. The positive rate of SARS-CoV IgG in cases who were not in chain of transmission was 12.59%(18/143) which was statistically significantly different from that of cases in chain of transmission, Chi square=199.64, P less than 0.001. During days 0-7,8-14,15-21,22-30 after onset, the cumulated positive rate of SARS-CoV IgG was 16.67%, 40.00%, 70.00% and 93.10%, respectively, then was kept at the level above 90% and lasted for 217 days. The cumulated positive rate of SARS-CoV IgM during days 0-7 after onset was the same to that of IgG. During days 8-14, 55.17% of cases had seroconversion for IgM which reached a peak (86.96%) during days 21-30. Then the rate rapidly declined.

CONCLUSIONMore than 94% of cases with SARS could produce IgG antibody when they were infected by SARS-CoV. Detecting SARS-CoV IgG could provide a diagnostic evidence for case confirmation. SARS-CoV IgG appeared as early as 7 days after onset and reached the peak at about weeks 4. Then the high rate of antibody was maintained for more than 6 months.

Antibodies, Viral ; blood ; Disease Transmission, Infectious ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; transmission

AIMTo study the anticancer effect of artesunate and its mechanism.

METHODSTo observe the effect of artesunate on the growth of solid tumor and the expression of PCNA, Bcl-2 and Bax genes in mice bearing H22 solid hepatic carcinoma.

RESULTSAfter administration of artesunate (ig, 300 mg.kg-1.d-1 x 7 d), growth of solid tumor was obviously inhibited, the tumor inhibitory rates were 49.1%, 48.7% and 46.6% and 46.6% in 3 experiments. The apoptosis of liver cancer cells were increased. Immunohistochemical staining showed that the number of Bcl-2 protein positive cells were decreased, but the number of Bax protein positive cells were increased. The PCNA positive cells were significantly lower than those in the control group.

CONCLUSIONArtesunate showed obvious anticancer activity on H22 hepatic carcinoma bearing mice and undergo apoptosis of liver cancer cells. The mechanism of anticancer effect of artesunate may be related to down-regulation of the expression of PCNA and Bcl-2 genes and up-regulation of the expression of Bax gene.

Animals ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Apoptosis ; Artemisinins ; therapeutic use ; Disease Models, Animal ; Female ; Gene Expression ; Hepatocytes ; metabolism ; pathology ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Proto-Oncogene Proteins ; biosynthesis ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Random Allocation ; Sesquiterpenes ; therapeutic use ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein

Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; SARS Virus ; immunology ; isolation & purification ; Seroepidemiologic Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; virology

OBJECTIVETo study the specific binding of the artificial clonal aryl hydrocarbon receptor translocator (ARNT) with the natural aryl hydrocarbon receptor (AhR) and the recolonization by polyclonal antibody. The dose-response relationship with tetrachlo-rodibenzo-dioxin (TCDD) was also studied to develop TCDD detection method and the binding degree related to dose response.

METHODS(1) The target genes including AhR-PAS, AhR-C and ARNT-PAS were amplified by RT-PCR by using the total RNA purified from the liver cells of C57BL/6J mice as templates to construct pGEX-5X1 recombinants. The recombinant plasmids were expressed in E. coli. (2) The rabbits were immuned by the clonal fusion proteins: AhR-PAS, AhR-C to prepare the polyclonal antibody. (3) The natural AhR from the hepatic cytosol of C57BL/6J mice was extracted. The artificial cloning expressed fusion protein:GST-ARNT-PAS and the natural AhR were incubated in different dose of TCDD. The quantity of the heterodimer through affinity adsorption and Western blots were measured.

RESULTS(1) The target proteins including AhR-PAS, AhR-C and ARNT-PAS were successfully cloned and expressed in E. coli. (2) The detection limit of polyclonal antibody AhR-PAS and AhR-C were 5 ng and 1 ng, respectively. (3) The total protein concentration prepared from the liver cells was 60.5 mg/ml. The artificial clonal protein ARNT-PAS could specifically bind to the natural AhR complex with the existence of TCDD. The detection limit of TCDD was 0.25 pmol which was 80 pg approximately.

CONCLUSIONA TCDD detection method based on the aryl hydrocarbon receptor system was established and the detection limit might reach pg grade.

Animals ; Cells, Cultured ; Limit of Detection ; Liver Extracts ; chemistry ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; analysis ; Rabbits ; Receptors, Aryl Hydrocarbon ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the epidemic situation and dominant strain of influenza in children with acute respiratory infection (ARI) during Flu season from Oct. 2005 to Mar. 2006 in Taiyuan.

METHODSMadin-darby canine kidney (MDCK) cell culture and hemagglutination inhibition (HI) assay were used to isolate and identify type A influenza viruses (H1N1 and H3N2) and B influenza viruses from clinical samples collected from outpatients who visited the Department of Pediatric because of ARI from Oct. 2005 to Mar. 2006. Oct. 2005 and Mar. 2006, we collected 415 blood samples from children and adults to detect the influenza virus antibody titers by HI test to exclude respiratory diseases.

RESULTS7 strains of H1N1 were isolated from 87 clinical specimens, with a positive rate of H1N1 as 8.04%. Out of 415 blood samples being collected, the positive rates and the geometric mean titer of H1N1 antibody Mar. 2006 were significantly higher in 0-3, 3-7 and 7-18 year-olds than Oct.2005.

CONCLUSIONH1N1 epidemic influenza did occur among children in winter and spring of 2005--2006 in Taiyuan city.

Adolescent ; Animals ; Antibodies, Viral ; blood ; Cell Line ; Child ; Child, Preschool ; China ; epidemiology ; Dogs ; Hemagglutination Inhibition Tests ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; isolation & purification ; Influenza B virus ; isolation & purification ; Influenza, Human ; epidemiology ; Population Surveillance

BACKGROUNDEarly diagnosis assumes a vital role in an effective treatment of Alzheimer's disease (AD). Most of the current studies can only make an AD diagnosis after the manifestation of typical clinical symptoms. The present study aimed to investigate typical and other biomarkers of AD to find a possible early biomarker.

METHODSA total of 14 5XFAD mice (at 3 and 6 months old), with 14 age-matched wild-type (WT) mice as control, were enrolled in this case-control study. Morris water maze test was performed to evaluate the cognitive function; buried food pellet test and olfactory maze test were employed to investigate the olfactory function; immunofluorescence to detect amyloid deposition and positron emission tomography to examine 2-deoxy-2-(18F) fluoro-D-glucose ([18F]-FDG) uptake in the hippocampus and cerebral cortex.

RESULTSWith the increasing age, cognitive performance (P = 0.0262) and olfactory function were significantly deteriorated (day 1 P = 0.0012, day 2 P = 0.0031, day 3 P = 0.0160, respectively) and the (18F)-FDG uptake was markedly decreased in multi-cerebral regions including the olfactory bulb (P < 0.0001), hippocampus (P = 0.0121), and cerebral cortex (P < 0.0001). Of note, in 3-month-old 5XFAD mice, a significant decline of (18F)-FDG uptake in the olfactory bulb was found when compared with that of age-matched WT mice (P = 0.023) while no significant difference was present when the uptakes in other cerebral regions were compared.

CONCLUSIONSThe decline of (18F)-FDG uptake in the olfactory bulb occurs earlier than other incidents, serving as an earlier in vivo biological marker of AD in 5XFAD mice and making early diagnosis of AD possibly.

Alzheimer Disease ; diagnosis ; Amyloid ; analysis ; Animals ; Animals, Genetically Modified ; Biomarkers ; analysis ; Fluorodeoxyglucose F18 ; metabolism ; Glucose ; metabolism ; Mice ; Olfactory Bulb ; metabolism ; Positron-Emission Tomography