Biomedical subjects

S Zhao

Publications and source records attributed to S Zhao.

At least 271 records · Page 15Linked to original sources

PubMed1999

[Changes in computed tomogram in cerebral parenchymal cysticercosis treated with albendazole].

AIM: To observe the changes in cerebral computed tomogram (CT) in cerebral parenchymal cysticercosis after albendazole treatment. METHODS: Cerebral CT scanning was conducted in 57 patients with cerebral cysticercosis in our hospital before, during and after albendazole treatment. RESULTS: Cerebral CT might be normal before onchospheres became cysticercariae in the brain. Small cystic lesions could turn into other CT signs of cerebral cysticercosis after albendazole treatment. CONCLUSION: Normal cerebral CT cannot rule out the disease. Small cystic lesions are the earliest pathological and active signs of the disease. Nodular focus occurs after the death of Cysticercus and calcification is the final outcome of the disease.

Adolescent↗

PubMed1999

[Effect of space flight on ultrastructure of chloroplast in Agastache rugosa].

OBJECTIVE: To substantiate the effects of space condition on medicinal plants, the seeds of Agastache rugosa were set up in retrievable satellites. METHODS: After returning to earth, the ultrastructural changes of chloroplasts were examined by means of electron microscope. RESULTS: Vesicles could be seen in the chloroplasts in the cells of earth controlled group, it was few and could not be found in the chloroplasts in the cells of weightless group and hit group. Some chloroplasts were degrading in the cells of weightless group, but this phenomenon was not observed in the cells of the other two groups. CONCLUSIONS: These results showed that space environment had some effect on the ultrastructure of chloroplasts in Agastache rugosa.

Agastache↗

PubMed1999

[Detection of human papillomavirus in laryngeal carcinoma with digoxigenin labelled probe prepared by polymerase chain reaction].

OBJECTIVE: To study the relationship between laryngeal carcinoma and human papillomavirus (HPV), and describe distribution, expression of genome types of HPV in laryngeal carcinoma. METHODS: We used a non-radioactive digoxigenin labeled probe prepared by polymerase chain reaction (PCR) to test the fresh tissue samples from 146 cases with different laryngeal lesions. The labelled probe of HPV consensus primers was obtained, which enabled detection of nine types of HPV DNAs that were HPV 6, 11, 16, 18, 31, 33, 35, 42 and 58. The genome types of HPV DNA positive cases were determined by using multiple primers PCR. RESULTS: The results turned out to be as follows: The HPV infection positive rate was 45.6%(31/68) for laryngeal carcinoma, 20.0% (3/15) for neck metastatic lymphnode, 11.8%(2/17) for precarcinomatous lesions and 6.3%(1/16) for vocal cord polypus. 15 cases of normal tissues adjacent to carcinoma and another 15 normal tissues opposite to the carcinoma were all HPV DNA negative. HPV16, 18 types are the main types of laryngeal carcinoma and HPV6, 11 types often appear in benign lesions of larynx. CONCLUSION: These results implicated that the occurrence and development of laryngeal carcinoma may be closely related to HPV infection.

DNA Probes, HPV↗

PubMed1999

[Simultaneous determination of tungsten, molybdenum and titanium by k-factor dual-wavelength standard addition method].

In this paper, a new method for simultaneous spectropthotometric determination of tungsten, molybdenum and titanium is proposed. The method is based on the difference of absorption spectra of the three elements forming coloured complexes with PF in acidic medium and in the presence of nonionic surfactant Tween-80. The absorbance of sample was measured at wavelengths of 503, 530 and 560 nm, and the data were treated by the principle of the k coefficients and dual-wavelength standard addition method. The method has been applied to the determination of tungsten, molybdenum and titanium in steels, and satisfactory results were obtained.

English Abstract↗

PubMed1998

Virulence genes of Shiga toxin-producing Escherichia coli isolated from food, animals and humans.

The presence of virulence genes, encoding enterohemorrhagic Escherichia coli (EHEC)-hemolysin (EHEC-hlyA), intimin (eae), and Shiga toxins 1 (stx1) and 2 (stx2), in 178 isolates of pathogenic E. coli, was determined using the polymerase chain reaction with primers specific for each virulence gene. The tested organisms were 120 isolates of E. coli O157:H7 from human patients, cattle, sheep and foods, 16 non-O157:H7 EHEC isolates from patients suffering from hemorrhagic colitis or hemolytic uremic syndrome, 15 non-O157:H7 Shiga toxin-producing E. coli (STEC) isolates from cattle and foods, 26 isolates of enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC) and enterotoxigenic E. coli (ETEC), and an E. coli K12 strain. Results revealed that all isolates of O157:H7 carried EHEC-hlyA, eae, and one or both stx genes; 15 of the 16 non-O157:H7 EHEC isolates had EHEC-hlyA, but all possessed eae and one or both stx genes; only seven of the 15 non-O157 STEC isolated from cattle and foods contained both EHEC-hlyA and eae genes. The EPEC, EIEC, ETEC, and the E. coli K12 strain did not carry these virulence genes, except eight EPEC isolates were positive for eae. Results suggest that a combination of EHEC-hlyA and eae genes could serve as markers to differentiate EHEC from less pathogenic STEC, and other pathogenic or non-pathogenic E. coli.

Adhesins, Bacterial↗

PubMed1998

Binding of the catalytic subunit of protein phosphatase-1 to the ryanodine-sensitive calcium release channel protein.

A number of studies have reported that the activity of the ryanodine-sensitive calcium release channel (ryanodine receptor) in the junctional sarcoplasmic reticulum of skeletal and cardiac muscle can be modulated by protein phosphorylation-dephosphorylation through activation of endogenous protein kinases and/or by addition of exogenous protein kinases and protein phosphatases. In this study, we have investigated the possibility that protein phosphatase-1 (PP1) is targeted to the junctional sarcoplasmic reticulum by the direct isolation of PP1-binding proteins on PP1-Sepharose affinity columns. The results show that the ryanodine receptor of both skeletal and cardiac muscle bind to this affinity support, and are released at supraphysiological salt concentrations in a relatively pure state. Reciprocal experiments demonstrated that PP1 binds to the immobilized muscle ryanodine receptor. The direct binding of PP1 to the ryanodine receptor was supported by the finding that tryptic fragments of the receptor were retained on PP1-Sepharose. The ability of PP1 to dephosphorylate the ryanodine receptor that was phosphorylated by protein kinase A was also demonstrated. These studies show that PP1 is targeted to the junctional sarcoplasmic reticulum by binding to the ryanodine receptor, and provide a biochemical basis for the possibility that PP1 may play a role in the regulation of calcium flux via protein phosphorylation-dephosphorylation mechanisms.

Animals↗

PubMed1998

Peptides that block hepatitis B virus assembly: analysis by cryomicroscopy, mutagenesis and transfection.

Peptides selected to bind to hepatitis B virus (HBV) core protein block interaction with the long viral surface antigen (L-HBsAg) in vitro. High resolution electron cryomicroscopy showed that one such peptide binds at the tips of the spikes of the core protein shell. The peptides contain two basic residues; changing either of two acidic residues at the spike tip to an alanine greatly reduced the binding affinity. Transfection of hepatoma cells with a replication-competent HBV plasmid gave significantly reduced production of virus in the presence of peptide, in a dose-dependent manner. These experiments show that the interaction of L-HBsAg with core particles is critical for HBV assembly, and give proof of principle for its disruption in vivo by small molecules.

Antiviral Agents↗

PubMed1998

Identification and characterization of the human HCG V gene product as a novel inhibitor of protein phosphatase-1.

The catalytic subunit of mammalian protein phosphatase-1 (PP1) is known to bind to a number of regulatory subunits, whose functions include the targeting of the catalytic subunit to the molecular proximity of its substrate proteins. In addition, PP1 is potently inhibited by several inhibitory polypeptides that include inhibitor-1 and inhibitor-2. In this study the yeast two-hybrid system was used to screen a human cDNA library for putative PP1-binding proteins. Ten putative positive clones were identified, one of which was found to be a partial cDNA of the hemochromatosis candidate gene V (HCG V) whose function was previously unknown. The full-length protein of 126 amino acid residues was expressed in Escherichia coli as a glutathione S-transferase fusion protein and also as a nonfusion protein. The recombinant protein inhibited recombinant and rabbit muscle protein phosphatase-1 with IC50s of ca. 1 nM, but did not inhibit PP2A. The term inhibitor-3 is proposed for this novel inhibitor. It is extremely hydrophilic, is heat stable, and behaves anomalously on SDS-PAGE with an apparent molecular mass of 23 kDa and on gel filtration with a relative molecular weight of 55 000, in contrast to its calculated molecular mass of 14 kDa. These characteristics are shared by the previously described protein phosphatase-1 inhibitor-2 and inhibitor-1 proteins.

Amino Acid Sequence↗

PubMed1998

Antiproliferative and antifibrotic effects of mimosine on adult cardiac fibroblasts.

Prolyl 4-hydroxylase catalyzes the hydroxylation of collagen pro-alpha chains for the deposition of cardiac collagen. The effect of prolyl 4-hydroxylase on synthesis and degradation of collagen was studied in cultured adult cardiac fibroblasts using mimosine, a prolyl 4-hydroxylase inhibitor. Mimosine inhibited [3H]thymidine incorporation in cultured fibroblasts in a dose-dependent manner (100-600 microM). Immunofluorescence in fibroblasts and biochemical detection of mature type I collagen in culture serum revealed a strong inhibition of synthesis and secretion of mature collagens, respectively, in the presence of 200 microM mimosine. Western blot analysis for procollagen was carried out in cultured fibroblasts, and 200 microM mimosine treatment was associated with increased intracellular accumulation of procollagen from 4.14+/-0.27 to 10. 19+/-0.37 (arbitrary units). Immunofluorescence studies confirmed a marked increase of intracellular procollagens in fibroblasts treated with mimosine, which suggests a loss of coordinated monomeric procollagen synthesis and secretion of triple helical mature collagens. Modest inhibition of collagen type I mRNA abundance was observed in mimosine-treated fibroblasts, whereas no effect was noted for mRNAs of collagen type III, alpha-prolyl 4-hydroxylase or beta-prolyl 4-hydroxylase when compared to untreated control values. Treatment of fibroblasts with 200 microM mimosine was associated with elevation of matrix metalloproteinase (MMP)-9 activity. The cytotoxicity of mimosine treatment was found minimal at the concentrations indicated above. Thus the antifibrotic effects induced by mimosine on cultured adult cardiac fibroblasts was associated with inhibition of prolyl 4-hydroxylase and diminished extracellular secretion of procollagen, despite the reactive elevation of intracellular procollagen synthesis. We suggest that specific inhibition of prolyl 4-hydroxylase may provide a novel therapeutic approach for the modulation of cardiac fibrosis.

Animals↗

PubMed1998

Ligation of cell surface CD38 protein with agonistic monoclonal antibody induces a cell growth signal in myeloid leukemia cells.

CD38 is expressed during early stages of differentiation in normal and leukemic myeloid cells. Recently, CD38 has been shown to participate in intracellular signal transduction pathways following its ligation with CD38-specific mAbs. In this study we report that ligation of CD38 by one such agonistic mAb (IB4) induced proliferation of cultured leukemic cells in vitro. In HL-60, KG-1A, NB4, and OCI-AML-3 myeloid leukemia cell lines, IB4 mAb induced an increase in the proliferating cell fraction as determined by cell number, clonogenic assay, and flow cytometric analysis. The presence of Ab caused a dose-dependent increase in the number of CFU and an increase in cell divisions. HL-60-Dox cells (a HL-60-doxorubicin-resistant cell line), which have no detectable CD38 expression, failed to respond to IB4 mAb. The effect of CD38 ligation on cell growth was also evaluated in freshly isolated leukemic cells from patients with acute myelogenous leukemia (AML). A significant increase in the proliferating cell fraction (S+G2M) was observed in 50% of the patients incubated with IB4 mAb. In five of the six AML patients, anti-CD38 mAb stimulated the proliferation of AML colony-forming cells. These results suggest that ligation of CD38 can induce the proliferation of leukemic cells and may play a role in the propagation of leukemic cell clones in certain cohorts of AML patients.

ADP-ribosyl Cyclase↗

PubMed1998

[Identification of a mutation hotspot in exon 8 of Wilson's disease gene by cycle sequencing].

OBJECTIVE: To screen for mutation hotspot of Wilson's disease (WD) gene in Chinese. METHODS: Cycle sequencing was used to detect mutation in exon 8 of WD gene in 30 patients with WD. RESULTS: The same missense mutation, Arg778Leu, was identified in 14 WD patients, four of which were homozygous and the others were heterozygous for this mutation. The frequency of this mutation was 30%. CONCLUSION: The codon 778(CGG-->CTG) of exon 8 in WD gene was one of mutation hotspots in Chinese.

Adolescent↗

PubMed1998

[Comparison of genotype and intellectual phenotype in untreated phenylketonuric children].

OBJECTIVE: The main feature of phenylketonuria(PKU) is mental retardation. Although classical PKU is defined as that the hepatic phenylalanine hydroxylase (PAH) activity ranges 0-1% of normal enzyme, the untreated PKU patients show a wide range of intellectual phenotype. This study sought to find the molecular basis of such variation of intellectual phenotype among PKU. METHODS: 45 classical PKU patients included in the research were screened for detecting six mutant alleles which were rather common among Chinese PKU patients, i.e. R243Q, R413P, Y204C, Y356X, W326X and R111X. PCR-ASO and PCR-SSCP techniques were used. The expression of those mutant PAH genes was analysed by methods of site-directed mutagencies. 27 PKU patients whose two mutant alleles were both defined were involved in this study. The IQ of these patients were tested by DDST system. RESULTS: Among 27 patients, 4/27 (15%) were mild retardation, 10/27(37%) were moderate, the severe mental retardation accounted for 12/27(44%). The relationship between genotype and intellectual phenotype in this group was examined. It was found that the intellectual phenotype of 8 patients were compatible with genotype but not well matched in 19 cases. The enzyme activity of Y204C expressed in vitro was 100%, but all 3 patients with Y204C/Y204C were severely mental retarded. Enzyme activities of R413P and Y356X were <3% and 0 respectively in expression analysis, but the patients in this group had mild or moderate mental retardation. CONCLUSION: Intellectual phenotype was not well matched with the genotype in classical PKU patients, so that genotype can not be used to predicte the intellectual phenotype in PKU patients.

Child↗

PubMed1998

[Screening mutations in phenylketonuria by means of nonradioactive reverse dot blot hybridization].

OBJECTIVE: To establish a simple, accurate and rapid method for screening of the mutant genes in phenylketonuria (PKU). METHODS: Four exons harboring the mutations, Y204C (exon6, E6), R243Q(E7), Y356X(E11) and R413P(E12), were amplified by polymerase chain reaction (PCR) with incorporation of biotinylated deoxynucleotide(biotinylated-11-dUTP or biotinylated-14-dCTP). Hybridization between immobilized allele-specific oligonucleotide probes and biotin-labelled amplified DNA was performed and nonradioactively detected by a colorimetric reaction using streptavidin-alkaline phosphatase. RESULTS: The methods of non-radioactive reverse dot blot hybridization were established to screen the mutations. We detected the genotypes of five PKU patients and found that three of them carried R243Q mutation and one of the three also carried Y356X mutation. These results were confirmed by PCR-single strand conformation polymorphism. CONCLUSION: This method is suitable for rapid screening for common mutations in Chinese PKU patients.

Genetic Testing↗

PubMed1998

[Chromosome assignment of a novel microsatellite polymorphic marker (D14S1435) by fluorescence in situ hybridization].

OBJECTIVE: Chromosome assignment of a novel microsatellite marker (D14S1435) containing CA repeats and showing 11 alleles and 0.85 of PIC value in Chinese population) isolated from microdissected human chromosome 14q24.3 DNA library by fluorescence in situ hybridization(FISH). METHODS: The polymorphic marker was used as probe to screen a human Lambda/DASH genome library, a positive clone was isolated and the recombinant phage DNA was extracted as primary material. The insert was recovered through BamH I digestion to complete and low melting point agarose gel electro- phoresis, after processed with Sau3A I the probe was prepared by linker- catch PCR method and the following fluorescence in situ hybridization(FISH) was conducted to metaphase chromosomes. RESULTS: this novel STR was precisely mapped to human chromosome 14q24.3, and this demonstrated the validity of isolating chromosome band-specific genetic markers from probe pool made by chromosome microdissection. CONCLUSION: Confirmed by chromosome fluorescence in situ hybridization, the novel STR obtained by chromosome microdissection, PCR and subcloning method was really from the micro- dissected region. This not only increases the numbers of genetic markers used for linkage analysis in the region,but also provides a novel valuable genetic marker for the gene diagnosis and gene cloning of the possible disease-causing genes located in the region of 14q24.3.

Chromosome Mapping↗

PubMed1998

[A study of genetic heterogeneity in Pfeiffer syndrome].

OBJECTIVE: To understand the molecular pathology of Pfeiffer syndrome. METHODS: DNA from peripheral blood was examined in 4 families with Pfeiffer syndrome by SSCP-sequence analyses and PCR-restriction enzyme digestion. RESULTS: The authors found the mutations of FGFR2 gene in two families, an A to G transition in the 3' acceptor splice site of intron 8 in a family, and Asp321 Ala substitution in exon 9 in another family. In addition, the mutation in exon 5 of FGFR1 gene (Pro252Arg) was found in a family. CONCLUSION: These findings reveal the genetic heterogeneity of Pfeiffer syndrome and can help one understand the molecular mechanism of the disease.

Acrocephalosyndactylia↗

PubMed1998

[Detection of SMN gene deletions in spinal muscular atrophy].

OBJECTIVE: Survival motor neuron gene(SMN) and neuronal apoptosis inhibitory protein gene (NAIP) have been identified as the candidates of progressive spinal muscular atrophy (SMA)-determining genes. The aims of this study were to investigate the absence of SMN gene exon 7 in Chinese SMA patients, to confirm the relationship between the deletion of the SMN and SMA further, and to establish methods for gene diagnosis and prenatal diagnosis of SMA. METHODS: PCR-SSCP with silver staining method was used to detect the genomic DNA of 37 SMA patients and 30 normal individuals for deletions of SMN exon 7. RESULTS: Homozygous deletion of the SMN exon 7 was identified in 86.7%(13/15) of type I SMA patients and 86.4%(19/22) of type II patients. In the 88 controls (including parents of patients and normal individuals), homozygous absence of SMA exon 7 was only found in a mother of a patient. CONCLUSION: The data support that homozygous absence of SMN exon 7 is strongly associated with SMA. The percentage of homozygous deletions in this study is almost as high as that reported by other researchers. This method is useful, reliable and effective for gene diagnosis and prenatal diagnosis of SMA.

Cyclic AMP Response Element-Binding Protein↗

PubMed1998

Expression of Gq alpha and PLC-beta in scar and border tissue in heart failure due to myocardial infarction.

BACKGROUND: Large transmural myocardial infarction (MI) leads to maladaptive cardiac remodeling and places patients at increased risk of congestive heart failure. Angiotensin II, endothelin, and alpha1-adrenergic receptor agonists are implicated in the development of cardiac hypertrophy, interstitial fibrosis, and heart failure after MI. Because these agonists are coupled to and activate Gq alpha protein in the heart, the aim of the present study was to investigate Gq alpha expression and function in cardiac remodeling and heart failure after MI. METHODS AND RESULTS: MI was produced in rats by ligation of the left coronary artery, and Gq alpha protein concentration, localization, and mRNA abundance were noted in surviving left ventricle remote from the infarct and in border and scar tissues from 8-week post-MI hearts with moderate heart failure. Immunohistochemical staining localized elevated Gq alpha expression in the scar and border tissues. Western analysis confirmed significant upregulation of Gq alpha proteins in these regions versus controls. Furthermore, Northern analysis revealed that the ratios of Gq alpha/GAPDH mRNA abundance in both scar and viable tissues from experimental hearts were significantly increased versus controls. Increased expression of phospholipase C (PLC)-beta1 and PLC-beta3 proteins was apparent in the scar and viable tissues after MI versus controls and is associated with increased PLC-beta1 activity in experimental hearts. Furthermore, inositol 1,4,5-tris-phosphate is significantly increased in the border and scar tissues compared with control values. CONCLUSIONS: Upregulation of the Gq alpha/PLC-beta pathway was observed in the viable, border, and scar tissues in post-MI hearts. Gq alpha and PLC-beta may play important roles in scar remodeling as well as cardiac hypertrophy and fibrosis of the surviving tissue in post-MI rat heart. It is suggested that the Gq alpha/PLC-beta pathway may provide a possible novel target for altering postinfarct remodeling.

Animals↗

PubMed1998

[Rapid selection of the gradually shortened DNA clones by using PCR amplification].

OBJECTIVE: To establish a simple technique with which the gradually shortened DNA clones can be selected rapidly and accurately so that the sequencing and identification of the full length cDNA or longer DNA fragments can be performed as quick as possible. METHODS: Primers were designed according to the sequence flanking the multiple cloning site of pGEM vector and were used to amplify the inserted fragments in a series of deletion sub-clones originated from a clone. with longer DNA insert fragment. These sub-clones which contain the gradually shortened fragments can be selected directly. This method was compared with the routine method in which the restriction endonuclease was used to digest DNA samples. RESULTS: A series of deletion sub-clones were identified accurately with this new method and the cycle sequencing could be performed on these PCR products as well. This method was characteristic of more accurate, more simple and less time-consuming, compared with the routine method. CONCLUSION: The method presented is very effective for rapid selection of the gradually shortened inserted fragments constructed in pGEM vector. It can greatly save time and materials, and the strategy in the report could also fit for selecting the clones containing gradually shortened fragments in other plasmid vectors.

DNA, Complementary↗