Creutzfeldt-Jakob disease in a Chinese patient with a novel seven extra-repeat insertion in PRNP.
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Biomedical subjects
Publications and source records attributed to Y J Guo.
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In our previous study, butein, a chalcone derivative, was found to be an inhibitor of tyrosine kinases and the inhibition was ATP-competitive. In this work, chalcone and seven chalcone derivatives were used to analyse the relationship between the structure of these compounds and their inhibition of tyrosine kinase activity. Three of chalcone derivatives, including butein, marein and phloretin, were found to have an ability to inhibit the tyrosine kinase activity of epidermal growth factor receptor (EGFR) in vitro. IC(50) was 8 microM for butein, 19 microM for marein and 25 microM for phloretin. The structural characterisations of these inhibitors suggest that the hydroxylations at C4 and C4' of these molecules may be required for them to act as EGFR tyrosine kinase inhibitors. The inhibition of EGF-induced EGFR tyrosine phosphorylation by butein was also observed in human hepatocellular carcinoma HepG2 cells, while marein and phloretin were inactive at the doses tested. Molecular modelling suggests that butein, marein and phloretin can be docked into the ATP binding pocket of EGFR. Hydrogen bonds and hydrophobic interaction appear to be important in the binding of these inhibitors to EGFR.
Apoptosis is a complex process involving a large array of genes and mutation of any of these genes may lead to malignancy formation. Re-acquirement of FasL by tumor cells may enable them to evade the surveillance of immune system and thus contributes to the growth of tumor. Apart from traditional therapies, inducing apoptosis of tumor cell by new methods employing death receptor ligands and making use of Fas counterattack is also being developed.
Peripheral blood lymphocytes from a single swine were stimulated with Concavadin A for 17 h, and the total RNA was isolated from it. Then, the mRNA specific for porcine IFN gamma was amplified by reverse transcription polymerase chain reaction. After sequencing, the IFN gamma gene has been successfully inserted into vector pJLA-503 and highly expressed in E. coli. Recombinant porcine IFN gamma expressed as inclusion body, which was dissolved in 7 mol/L guanidine chloride and subsequently renatured by dilution in refolding buffer containing 0.5 mol/L L-arginine. In order to obtain pure protein, the renatured IFN gamma was purified by the chromatographies of SP-Sepharose FF and Sephacryl S-200 HR. As a result, the final pure product can been seen as a single band in SDS-PAGE, and the cytokine activity was verified by inhibiting the cytopathic effect.
In previous work, the cDNA encoding Cysticercus cellulose annexin32 has been cloned. With PCR method, two different restriction Sites were added to each end of the cDNA respectively. Then, the cDNA was inserted into prokaryotic expression vector pJLA-503. After inducing, most foreign protein was expressed in soluble form, which was up to 35% of the total protein of the bacteria. Subsequently, the recombinant Annexin32 was purified with (NH4)2SO4 stepwise precipitation, DEAE-Sepharose FF and Sephacryl S-200 HR chromatography. The final pure protein can been shown as a single band in SDS-PAGE, and the biological activity was verified by Western blot and anticoagulation activity assay.
OBJECTIVE: To analyse the immunostimulatory activity of CpG sequences in cysticercus cellulosae paramyosin (also named Antigen B, AgB) cDNA. METHODS: C57BL/6 mice were immunized with pcDNA3-AgB plasmid, pcDNA3-AgB' (CpG sequences were mutated), pcDNA3 or AgB protein and two weeks later, immune response was assayed by ELISA. RESULTS: IgG and IgG2a were detectable at week 2 after immunization and continually increased until week 4. The antibody levels elicited by pcDNA3-AgB were significantly higher(P < 0.05) than those elicited by others. CONCLUSION: After pcDNA3-AgB plasmid inoculation, the immune response of mouse was elicited not only by the AgB protein but also by the CpG immunostimulatory sequences in the AgB cDNA.
OBJECTIVE: To compare the chemical composition and bioactivity of polypeptides(PPs) isolated from velvet antlers of sika deer (Cervus nippon Temminck) and red deer (Cervus elaphus Linnaeus). METHOD: The two kind of polypeptides were isolated from the above mentioned velvet antlers with same technology. The chemical composition was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and MALDI-TOF mass spectrometry. Stimulant activity of cells proliferation was measured by [3H] TdR incorporation into DNA. RESULT: The graphs of SDS-PAGE and MALDI-TOF MS of velvet antler polypeptides (VAPPs) from Chinese and New zealand red deer were very similar, but there were obvious difference in respect of graph between sika deer and red deer. VAPPs 25-50 mg.L-1 showed marked proliferation-promoting activity for rabbit costed chondrocytes, either sika deer or red deer. However, the activity of sika deer VAPPs 12.5 mg.L-1 for epidermal cells was weaker than that of red deer (12.5 mg.L-1). CONCLUSION: The chemical property and bioactivity of VAPPs from sika deer and red deer are significantly different.
OBJECTIVE: To establish the determination method of Shikimic acid. METHOD: Using HPLC method as the determination method. The separation was performed in a SiO2-NH2 column with a mobile phase of Acetonitrile-2% H3PO4 water solution (95:5); The sample wavelength was 213 nm, reference wavelength 300 nm. RESULT: The average collection was 98.5%, RSD 1.67% (n = 5). CONCLUSION: This method is suitable for the determination of Shkimic acid in herb medicines and preparation containing shikimic acid.
The reported transmission of avian H9N2 influenza viruses to humans and the isolation of these viruses from Hong Kong poultry markets lend urgency to studies of their ecology and pathogenicity. We found that H9N2 viruses from North America differ from those of Asia. The North American viruses, which infect primarily domestic turkeys, replicated poorly in inoculated chickens. Phylogenetic analysis of the hemagglutinin and nucleoprotein genes indicated that the Asian H9N2 influenza viruses could be divided into three sublineages. Initial biological characterization of at least one virus from each lineage was done in animals. Early isolates of one lineage (A/Chicken/Beijing/1/94, H9N2) caused as high as 80% mortality rates in inoculated chickens, whereas all other strains were nonpathogenic. Sequence analysis showed that some isolates, including the pathogenic isolate, had one additional basic amino acid (A-R/K-S-S-R-) at the hemagglutinin cleavage site. Later isolates of the same lineage (A/Chicken/Hong Kong/G9/97, H9N2) that contains the PB1 and PB2 genes similar to Hong Kong/97 H5N1 viruses replicated in chickens, ducks, mice, and pigs but were pathogenic only in mice. A/Quail/Hong Kong/G1/97 (H9N2), from a second lineage that possesses the replicative complex similar to Hong Kong/97 H5N1 virus, replicated in chickens and ducks without producing disease signs, was pathogenic in mice, and spread to the brain without adaptation. Examples of the third Asian H9N2 sublineage (A/Chicken/Korea/323/96, Duck/Hong Kong/Y439/97) replicated in chickens, ducks, and mice without producing disease signs. The available evidence supports the notion of differences in pathogenicity of H9N2 viruses in the different lineages and suggests that viruses possessing genome segments similar to 1997 H5N1-like viruses are potentially pathogenic in mammals.
Piperidine alkaloids compose most of the venom of the red imported fire ant, Solenopsis invicta, and we examined how six of these alkaloids varied across worker size and age. In a colony sampled intensively, the relative abundance of each alkaloid was highly correlated with worker size with one exception, and ratios of saturated to unsaturated alkaloids were positively correlated with worker size. Similarly, both the abundance and ratios of alkaloids differed significantly between the small and large workers sampled from colonies across Texas, USA. Young and old workers produced less venom than ants of intermediate age (3-7 weeks), and ratios of saturated to unsaturated alkaloids increased significantly with worker age. The differences in venom composition correspond to the size- and age-based functional roles of workers.
OBJECTIVE: To investigate the chemical constituents of Phyllanthus urinaria. METHOD: Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical and spectral analyses. RESULT AND CONCLUSION: Fourteen compounds were isolated and seven of them were identified as corilagin(I), rutin(II), brevifolincarboxylic acid(VI), isostrictiniin(IX), geraniin (X), gallic acid(XI) and ellagic acid (XII). Compound VI was found from P. urinaria for the first time and compound IX was found from genus Phyllanthus for the first time.
OBJECTIVE: To investigate the chemical constituents of Phyllanthus urinaria. METHOD: Various chromatographic techniques were employed for isolation and purification. The structures were elucidated by spectral analyses. RESULT AND CONCLUSION: A novel polyphenolic compound was isolated and named phyllanthusin F.
Formation of the mammalian secondary palate is a highly regulated and complex process whose impairment often results in cleft palate, a common birth defect in both humans and animals. Loss-of-function analysis has linked a growing number of genes to this process. Here we report that Lhx8, a recently identified LIM homeobox gene, is expressed in the mesenchyme of the mouse palatal structures throughout their development. To test the function of Lhx8 in vivo, we generated a mutant mouse with a targeted deletion of the Lhx8 gene. Our analysis of the mutant animals revealed a crucial role for Lhx8 in palatogenesis. In Lhx8 homozygous mutant embryos, the bilateral primordial palatal shelves formed and elevated normally, but they often failed to make contact and to fuse properly, resulting in a cleft secondary palate. Because development of other craniofacial structures appeared normal, the impaired palatal formation in Lhx8-mutant mice was most likely caused by an intrinsic primary defect in the mesenchyme of the palatal shelves. The cleft palate phenotype observed in Lhx8-mutant mice suggests that Lhx8 is a candidate gene for the isolated nonsyndromic form of cleft palate in humans.
AIM: To study the effects of velvet antler (VA) total polypeptides (VATP) and VA polypeptides, VAP-A, VAP-B, and VAP-C on proliferation of chondrocytes and osteoblast precusors. METHODS: Chondrocytes (rabbit and human fetus) and osteoblast precusors (chick embryo) were incubated in the culture medium containing VATP or VAP-A, VAP-B, and VAP-C. [3H]TdR incorporation into DNA was measured. Fracture healing-promoting action of VATP was determined in rats. RESULTS: VATP 50-200 mg.L-1 and VAP-B 12.5, 25, and 50 mg.L-1 showed most marked proliferation-promoting activity for rabbit costed chondrocytes and increased incorporation of [3H]TdR from (73 +/- 9) Bq (control group) to (272 +/- 55), (327 +/- 38), and (415 +/- 32) Bq, respectively (P < 0.01). The activity of VAP-A was weaker than that of VAP-B, and VAP-C had no activity. VATP 10 and 20 mg.kg-1 by local injection into the cross-section fracture area accelerated healing of radial fracture. The healing rate of VATP-treated group was higher (75%) than that of control group (25%) (P < 0.05). CONCLUSION: VATP accelerated fracture healing by stimulating proliferation of chondrocytes and osteoblast precursors.
AIM: To study the effects of shikimic acid (SA) on focal cerebral ischemic injury after middle cerebral artery thrombosis (MCAT). METHODS: Thrombosis was induced by FeCl3 in middle cerebral artery of rats. The influences of SA on neurologic deficit (ND), infarct size (IS), brain edema, and cerebral blood flow (CBF) in ischemic region were observed. RESULTS: SA 25 and 50 mg.kg-1 i.p. for 3 d before MCAT attenuated ND, and reduced IS by 51% and 42%; and decreased brain water content from 80.7% to 79.8% and 79.9%; and increased CBF after ischemia from 50.2% of the preischemic level to 75.5% and 73.3%, respectively. In pathologic examination, there was much less thrombosis in MCA in the rat with the pretreatment by SA 25 mg.kg-1. The extent of brain ischemia was much less than that of control. CONCLUSIONS: SA reduced focal cerebral ischemic injury induced by middle cerebral artery thrombosis.
AIM: To study the inhibitory effect of chlorpromazine (Chl), verapamil, and aspirin on hepatocyte apoptosis induced by the cessation of phenobarbital (Phe) treatment in mice. METHODS: Liver DNA content, ratio of liver weight/body weight, DNA fragmentation, DNA electrophoresis, the end-labeling test (TUNEL), and the morphologic changes of liver cells as indices of liver mass and hepatocyte apoptosis were applied to investigate (1) the kinetic process of hepatocyte proliferation induced by Phe 75 mg.kg-1 i.p. and the regression of hyperplastic liver caused by withdrawal of Phe in mice, (2) the effect of Chl 25 mg.kg-1, verapamil 50 mg.kg-1 or aspirin 60 mg.kg-1 i.p. on mouse hepatocyte apoptosis, and (3) the time course of effects of Chl on the regression of liver size and DNA fragmentation content after withdrawal of Phe. RESULTS: The process of hepatocyte proliferation and regression induced by administration and withdrawal of Phe in mice consisted of 4 phases: proliferation, plateau, rapid regression, and slow regression phases. In the rapid regression phase, the typic changes of hepatocyte apoptosis were found, which was prevented in early period by the Ca(2+)-calmodulin antagonist Chl, but not by verapamil or aspirin. CONCLUSION: The Ca(2+)-calmodulin played an important role in the hepatocyte apoptosis caused by withdrawal of Phe.
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