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Biomedical subjects

S Li

Publications and source records attributed to S Li.

At least 1,477 records · Page 82Linked to original sources

[Interruption of perinatal transmission of HBV by Chinese recombinant hepatitis B vaccine].

Sixty-two infants who were born from HBsAg and HBeAg both positive mothers were vaccinated with 20 micrograms/dose recombinant hepatitis B vaccine, which was made in Beijing Institute of Biologic Products (Vaccinia virus recombinant hepatitis B vaccine) and Changchun Institute of Biologic Products (Chinese hamster ovary cells recombinant hepatitis B vaccine), at 0, 1, 6 months. All the babies were followed up for 9 or 12 months. The results showed that the protective rates of Beijing vaccine and Changchun vaccine were 79.2% and 80.2%, respectively, the positive rates of anti-HBs were 78.8% and 79.3%, respectively. Local or systematic reactions were not found in all the babies 3 days after injection. These results suggested that the Chinese recombinant hepatitis B vaccine was safe and effective. It was better than the plasma vaccine with the same dose.

Female↗

[Pathologic characteristics of diffuse alveolar damage induced by paraquat in rat lungs].

Pathologic characteristics and events of paraquat (PQ) induced diffuse alveolar damage were observed by pathohistologic and ultrastructural studies of the lungs of rats, which were given a single intraperitoneal injection of PQ 25mg/kg body weight and sacrificed 6 hours to 45 days later. Results showed that the capillary endothelial and type I epthelial cells were mainly damaged, and associated with interstitial oedema, haemorrhage and hyaline membrane formation of the alveoli, and accentuated alveolitis. The lesions were located in the alveolar structural units and very diffuse in distribution. When the pulmonary damage became irreversible, it then led to fibrosis.

Animals↗

[Cytogenetic studies on a human cecum adenocarcinoma cell line (HCe-8693)].

Human undifferentiated cecum adenocarcinoma cell line (HCe-8693) was established by Zhejiang Cancer Hospital in 1986. This paper reports the results of cytogenetic studies on the 23rd (early) and 125th (late) passages of this cell line. The numerical distribution range of chromosomes was rather narrow, showing a near diploid karyotype, and no significant difference was observed between early and late passages. The modal number of chromosomes was 49. G-banding karyotypic analysis revealed that the karyotype of this cell line was close to that of normal diploid cells, with few, easily identified abnormal chromosomes. The most significant changes were a distal deletion of the short arm of chromosome No. 1 and a trisomy of chromosome No. 13. The morphological characteristics and possible correlation with pathogenesis of colorectal cancer were briefly discussed.

Adenocarcinoma↗

[Difference in the distribution and content of nucleolar protein B23 between interphase and mitotic HeLa cells].

Protein B23 is one of the two major protein components in eukaryocytic nucleolus. Previous studies indicate that the cellular content and distribution of protein B23 are correlated with the synthetic rate of ribosomal RNA (rRNA) and cell growth. It is more abundant in tumor and growing cells than in normal resting cells. It may translocate from nucleolus to nucleoplasm after serum deprivation, or after being treated with RNA inhibitors such as actinomycin D, toyocamycin, or doxorubicin. In order to search for the role of protein B23 in cell division, using an anti-B23 monoclonal antibody, we detect the cellular content and distribution of protein B23 in interphase and mitotic HeLa cells, respectively. Two microtubule drugs, colcemid and taxol, are used to arrest the mitotic cells. The results show the cellular content of protein B23 in mitotic cells is much higher than that in interphase cells. The intracellular distribution of protein B23 in mitotic cells is distinguished from that in interphase cells too.

Blotting, Western↗

Age-related alterations of proteoglycan in mouse tracheal cartilage matrix: an electron histochemical analysis with the cationic dye of polyethyleneimine.

Age-related alterations of the proteoglycans (PGs) contents and size were observed in the tracheal cartilage matrix of ddY mice at various ages with the cationic dye of polyethyleneimine (PEI). In young animals (aged 1 day to 1 month after birth), the PGs content and size increased, resulting in the formation of the territorial matrix (TM) and the decrease of the space between the PEI aggregates and the collagen fibers. In 1 month old mice, the TM could be distinguished from the interterritorial matrix (ITM). In adult mice (from 2 to 6 months), the PGs content and size kept stable, but the rearrangement of the PGs and collagen fibers was observed. From 6 months of age on, the decrease of PGs content and size occurred mainly in the ITM. The developing stopping sign was recognized in 1 month and the senile sign from 6 months in the ddY mice as for the complete formation of the TM in 1 month and the decrease of the PGs content and size from 6 months. These results demonstrated the age-related alteration of PGs in the ultrastructural level first and could contribute to the aging study of the tracheal cartilage.

Aging↗

An E2F binding sequence negatively regulates the response of the insulin-like growth factor 1 (IGF-I) promoter to simian virus 40T antigen and to serum.

The promoter of the Insulin-like growth factor I (IGF-I) gene is activated by the Simian Virus 40 large T antigen (SVLT), and one of the elements responding to SVLT activation has been localized to a short 124 bp immediately upstream of the first initiation of transcription site. This short promoter contains an E2F binding site, that, in gel shifts, binds a protein complex, but only when the promoter activity is reduced or absent. A mutation in the E2F binding site deregulates the activity of the promoter, which becomes active even in those conditions in which the wild type promoter is inactive. By using antibodies in gel retardation analyses, we can show that the different protein complexes include, at least, the following proteins: E2F, cyclin A and p107. We conclude that the short IGF-I promoter is negatively regulated by an E2F binding site that complexes with several proteins. Our data suggest that disaggregation of these complexes by the action of SVLT (or other activators) increases expression from the promoter, thus establishing a link between the regulation of cell proliferation by growth factors and the E2F-associated proteins.

3T3 Cells↗

[Computer assisted studies on the structural requirements for the mutagenicity in Salmonella reversion assay for organophosphorus pesticides].

Computer assisted molecular fragment evaluation (CAMFE) program has been established and applied to the data set analysis of the mutagenicity of 59 organophosphorus pesticides (OPPs) in Salmonella reversion assay. The results favour the hypothesis that the mutagenic properties of OPPs lie in their alkylation to DNA mainly by the molecular fragments containing methyl groups rather than those containing ethyl groups. Oxygen atom in phosphoryl group may prompt the mutagenic ability induced by OPPs; However, the mutagenic ability may be decreased by sulfur atom, the substitute of oxygen atom at the same site in PPs structure. It was suggested that, among the most relevant fragments, the methoxyphosphinyl group appears as the common structural subunit responsible for the activities detected in the Salmonella reversion assay, while the ethyl thionophosphinyl group appears as the descriptor of negative results.

Insecticides↗

[Comparative study of single strand conformation polymorphism of 4.5S RNA gene in enterobacteria].

A recently developed technique, non-isotopic single strand conformation polymorphism analysis (PCR-SSCP), was applied to study the conserved feature of 4.5S RNA gene in enterobacteria. The 4.5S RNA gene was amplified by the polymerase chain reaction, using the template DNA extracted respectively from five strains of Escherichia coli and three strains of different genera in Enterobacteriaceae, i.e. Proteus vulgaris, Serratia marcescens and Enterobacter aerogenes. The PCR products were then carried out SSCP analysis. The experimental results showed that there seemed to be no detectable differences in the size and single strand conformation of 4.5S RNA genes from above strains, except the negative strand conformation of Enterobacter aerogenes. Thus it can be seen that the secondary structures of 4.5S RNA gene in enterobacteria are quite conservative.

Base Sequence↗

A study of c-myc oncogene expression and amplification in colorectal cancer.

Expression of c-myc oncogene transcripts in colorectal neoplasia was studied in paraffin embedded tissue sections from 25 patients undergoing surgery and from the rectal carcinoma cell line HR-8348 by using in situ hybridization, and its amplification was investigated in tumor and normal mucosa tissue from 25 coloproctomy samples by slot blot hybridization. Overexpression of this gene was seen in 78% (7/9) of the benign adenomas and 91% (20/22) of the malignancies sampled. There was no significant correlation between overexpression and the histologic type or grade, and no significant relationship between the level of expression and clinical stage was found, although overexpression was apparently more common in tumors with metastasis. Amplification of the gene was found in 0 of 4 benign adenomas and 7 of 22 malignancies. No obvious correlation was found between amplification and histological type or grade, though amplification was more frequent in tumors with metastasis. Amplification was also found in 2 adenomas with malignant change. The results suggest that multiple factors are involved in the progression of colorectal cancer, and in situ hybridization with a nonradiolabeled probe is useful in the detection of gene expression.

Adenocarcinoma↗

Molecular cloning and expression of a rat kappa opioid receptor.

At least three types of opioid receptors have been identified in the nervous system. In this paper we report molecular cloning and expression of a rat kappa opioid receptor. PCR was performed on double-stranded cDNA derived from poly(A)+ RNA of the rat striatum with primers similar to those of Libert and co-workers [Libert, Parmentier, Lefort, Dinsart, Van Sande, Maenhaut, Simons, Dumont and Vassart (1989) Science 244, 569-572]. One of the PCR products, which had 65% sequence similarity to the mouse delta opioid receptor, was used to screen a rat striatum cDNA library. Two positive clones were isolated and found to be identical. The clone had a 2.1-kb insert, which was termed RKOR-1. RKOR-1 has an open reading frame of 1140 bp and encodes a 380-amino-acid protein. Hydropathy analysis indicates that RKOR-1 has seven putative transmembrane domains with short intra- and extra-cellular loops. Membranes of Cos-7 cells transfected with RKOR-1 exhibited high specific binding for [3H]diprenorphine ([3H]DIP), a non-selective opioid ligand. Naloxone inhibited [3H]DIP binding with stereospecificity. [3H]DIP binding was potently inhibited by selective kappa opioid ligands, with Ki values in the nanomolar or subnanomolar range, but much less potently inhibited by drugs selective for mu or delta receptors. Thus, RKOR-1 represents an opioid receptor with kappa characteristics.

Amino Acid Sequence↗

Raf-1 protein kinase activates the NF-kappa B transcription factor by dissociating the cytoplasmic NF-kappa B-I kappa B complex.

Addition of mitogenic growth factors to quiescent cells triggers complex signal transduction cascades that result in the reprogramming of gene expression and entry into the cell cycle. We have found that an oncogenic variant of the c-Raf-1 protein kinase stimulated the expression of promoters containing NF-kappa B binding sites. In situ immunofluorescence analysis revealed elevated nuclear levels of the p65 subunit of NF-kappa B in v-raf-transformed NIH 3T3 cells. Incubation of HeLa cell cytoplasmic extracts with a purified recombinant glutathione S-transferase-raf fusion protein in the presence of ATP released active NF-kappa B that could be detected by electrophoretic gel mobility shift assay. Coincubation of purified recombinant I kappa B and glutathione S-transferase-raf in the presence of ATP resulted in the phosphorylation of I kappa B. Coexpression of GAL4 (activation domain)-I kappa B and GAL4 (DNA-binding domain)-raf fusion proteins in yeast resulted in stimulation of a GAL4-responsive reporter gene, indicating that I kappa B and Raf interact physically in vivo. These results indicate that the Raf-1 kinase functions in signal transduction in part by activating the NF-kappa B transcription factor by phosphorylating I kappa B in the cytoplasmic I kappa B-NF-kappa B complex to release active NF-kappa B.

3T3 Cells↗

Reductive dehalogenation by cytochrome P450CAM: substrate binding and catalysis.

Biological reductive dehalogenation reactions are important in environmental detoxification of organohalides. Only scarce information is available on the enzymology underlying these reactions. Cytochrome P450CAM with a known X-ray structure and well-studied oxygenase reaction cycle, has been studied for its ability to reduce carbon-halogen bonds under anaerobic conditions. The reductive reactions functioned with NADH and the physiological electron-transfer proteins or by using artificial electron donors to reduce cytochrome P450CAM. Halogenated methane and ethane substrates were transformed by a two-electron reduction and subsequent protonation, beta-elimination, or alpha-elimination to yield alkanes, alkene, or carbene-derived products, respectively. Halogenated substrates bound to the camphor binding site as indicated by saturable changes in the Fe(III)-heme spin state upon substrate addition. Hexachloromethane was bound with a dissociation constant (KD) of 0.7 microM and caused > 95% shift from low- to high-spin iron. Ethanes bearing fewer chlorine substituents were bound with increasing dissociation constants and gave lesser degrees of iron spin-state change. Camphor competitively inhibited hexachloroethane reduction with an inhibitor constant (KI) similar to the dissociation constant for camphor (KI = KD = 0.9 microM). Rate determinations with pentachloroethane indicated a 100-fold higher enzyme V/K compared to the second-order rate constant for hematin free in solution. These studies on substrate binding and catalysis will help reveal how biological systems enzymatically reduce carbon-halogen bonds in the environment.

Camphor 5-Monooxygenase↗

Mixture analysis of pursuit eye-tracking dysfunction in schizophrenia.

Results of two recent studies suggest that a distinct subgroup of schizophrenic patients and their relatives have particularly deviant eye tracking. Such heterogeneity could be of considerable importance, as it may indicate significant pathophysiologic or etiologic heterogeneity in schizophrenia. An analysis of 101 consecutive-admission schizophrenic patients confirmed the existence of two distinct subgroups of patients with higher and lower levels of spatial [root mean square (RMS)] eye-tracking error. However, there was no heterogeneity in the disturbance of pursuit eye movements. Anticipatory saccades, which by definition add very large amounts of spatial tracking error, were more frequent in the "high" RMS error group. Rates of anticipatory saccades were similar in the "low" RMS error patient group and normal controls, and there was no heterogeneity in the expression of anticipatory saccades. Apparent heterogeneity in global indices of eye-tracking impairment in schizophrenia appears to be a measurement artifact reflecting the powerful influence of anticipatory saccades on global performance indices, rather than true heterogeneity in the expression of any specific eye movement abnormality.

Adolescent↗

The anticodon and discriminator base are important for aminoacylation of Escherichia coli tRNA(Asn).

A gel shift assay that distinguishes the aminoacylated form from the deacylated form of tRNAs was used to study the requirements for aminoacylation of Escherichia coli tRNA(Asn) in vivo. tRNA(Asn) derivatives containing single base changes in their anticodons or discriminator bases were constructed, and the extent of in vivo aminoacylation was determined directly. Substitution of U35 with C35 or U36 with C36 abolished aminoacylation of the tRNA. Substitution of G34 with C34 converted tRNA(Asn) into a lysine acceptor. Thus, each of the anticodon nucleotides are important for aminoacylation of tRNA(Asn). Substitution of discriminator base G73 with A73 affected the extent of aminoacylation in vivo indicating that the discriminator base also contributes to aminoacylation of tRNA(Asn).

Acylation↗

Myogenin gene disruption results in perinatal lethality because of severe muscle defect.

Myogenin is a member of the basic helix-loop-helix (bHLH) gene family and converts multipotential mesodermal cells to myoblasts. The four members of the myoD family show unique spatio-temporal expression patterns and therefore may have different functions during myogenesis. Here we inactivate the myogenin gene in order to understand its role in myogenesis. Homozygous mutations are lethal perinatally owing to the resulting major defects in skeletal muscle. The extent of disorganization of muscle tissue differs in three regions. In the latero-ventral body wall, most cells, including myogenic cells, disappear and there is rapid accretion of fluid. In the limbs, cells of the myogenic lineage exist, but they are severely disrupted, and some of them are mono-nucleate with properties of myoblasts. In contrast, there are many axial, intercostal and back muscle fibres to be seen, although fibres are mainly disorganized and Z-lines are not present in most myofibrils. These findings are evidence that myogenin is crucial for muscle development in utero and demonstrate that other members of the myogenic gene family cannot compensate for the defect.

Animals↗

Elimination of band compression in sequencing gels by the use of N4-methyl-2'-deoxycytidine 5'-triphosphate.

Taq DNA polymerase, Sequenase, and the large fragment of E.coli polymerase I effectively utilize N4-methyl-2'-deoxycytidine 5'-triphosphate (N4-methyl-dCTP) in the place of dCTP in dideoxynucleotide terminator sequencing reactions on single-stranded templates. When the resulting fragment mixtures are resolved on sequencing gels, they are found to be free of band compressions even in cases where such compressions remain unresolved by the substitution of 7-deaza-dGTP for dGTP. Sequencing reactions using N4-methyl-dCTP instead of dCTP are somewhat more prone to false stops than are sequencing reactions using 7-deaza-dGTP instead of dGTP; this difference is more pronounced when sequencing with Sequenase at 37 degrees C than when sequencing with Taq DNA polymerase at 72 degrees C. For the three polymerases investigated, replacement of dCTP by N4-methyl-dCTP does not fundamentally change the characteristic variations in band intensities seen in the C-lane. N4-methyl-dCTP can also be used for sequencing double-stranded DNA and for DNA amplification by the polymerase chain reaction.

Base Sequence↗

Priming with recombinant influenza virus followed by administration of recombinant vaccinia virus induces CD8+ T-cell-mediated protective immunity against malaria.

Live vectors expressing foreign antigens have been used to induce immunity against several pathogens. However, for the virulent rodent malaria parasite Plasmodium yoelii, the use of recombinant vaccinia virus, pseudorabies virus, or Salmonella, expressing the circumsporozoite protein of this parasite, failed to induce protection. We generated a recombinant influenza virus expressing an epitope from the circumsporozoite protein of P. yoelii known to be recognized by CD8+ T cells and demonstrated that this vector induced class I major histocompatibility complex-restricted cytotoxic T cells against this foreign epitope. Immunization of mice with this recombinant influenza virus, followed by a recombinant vaccinia virus expressing the entire circumsporozoite protein, induced protective immunity against sporozoite-induced malaria. The sequence of immunization appears to be crucial, since a primer injection with recombinant vaccinia virus, followed by a booster injection with recombinant influenza virus, failed to induce protection. The protection induced by immunization with these recombinant viruses is mostly mediated by CD8+ T cells, as treatment of mice with anti-CD8 monoclonal antibody abolishes the anti-malarial immunity. The use of different live vectors for primer and booster injections has a synergistic effect on the immune response and might represent an effective general strategy for eliciting protective immune responses to key antigens of microbial pathogens.

Amino Acid Sequence↗

A selective defect in IgG2b switching as a result of targeted mutation of the I gamma 2b promoter and exon.

LPS stimulation of B lymphocytes induces germline transcription of and subsequent switching to the gamma 2b gene. Mature germline transcripts contain an I exon (non-coding) spliced to the C gamma 2b exons. To investigate the role of germline transcription and/or transcripts in heavy chain class switching, we have replaced the germline I gamma 2b promoter and I exon in ES cells with an expressed neomycin resistance gene. The mutated chromosome retains the downstream target sequence for switch recombination (S regions) and all sequences necessary for expression of a switched gamma 2b gene. Wild-type or mutant ES cells were injected into RAG-2 deficient blastocysts to generate somatic chimeras in which all lymphocytes were ES-cell derived. Chimeras derived from injection of heterozygous mutant ES cells had normal levels of serum IgG2b, but their splenic B cells showed a partial decrease in ability to switch to gamma 2b. Strikingly, B lymphocytes from chimeras derived by injection of homozygous mutant ES cells were deficient in IgG2b production both in vivo and in vitro, but normal with respect to production of other Ig heavy chain isotypes. Additional studies demonstrated that lack of ability to produce IgG2b by the mutant B cells correlated with lack of germline transcription and resulted from a specific defect in class-switch recombination to S gamma 2b. Together, these studies demonstrate that the I region is an important regulatory element for control of class-switch recombination.

Animals↗