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Biomedical subjects

C Lin

Publications and source records attributed to C Lin.

At least 145 records · Page 8Linked to original sources
PubMed1999

Deletion and nonsense mutations of the connexin 32 gene associated with Charcot-Marie-Tooth disease.

Two patients with a mild to moderate phenotype of Charcot-Marie-Tooth disease were identified to carry the mutations of the connexin (Cx) 32 gene. One of the patient had a novel nonsense mutation of tryptophan at amino acid 132 and the other had a deletion of the Cx 32 gene. Our study indicated that a loss of Cx 32 function contributes to a major pathogenesis of X-linked Charcot-Marie-Tooth disease.

Adult↗
PubMed1999

Capillary zone electrophoresis: an additional technique for the identification of hemoglobin variants.

Two capillary zone electrophoresis kits (Hb A2 and Hb A1c) were tested for confirmation and identification of hemoglobin variants. The capillary zone electrophoresis experiments were performed at pH 4.7 (Hb A1c kit) and 8.7 (Hb A2 kit) in a 24 cm uncoated fused silica capillary tube (25 microm I.D.). Normal hemoglobins and common hemoglobin variants, including Hbs S, D-Punjab, C, E, O-Arab, and G-Philadelphia, were successfully separated by both methods within a few minutes. Both systems provided completely different elution profiles of normal and abnormal hemoglobin fractions tested and were complementary. The inter-assay coefficient of variations of the migration times of hemoglobin variants were less than 1.0 and 1.3% by the Hb A2 and Hb A1c, respectively. This permits a higher resolution of some hemoglobin variants in low concentrations, like Hb S in newborns, compared with conventional electrophoresis methods. The present capillary zone electrophoresis methods are sensitive, rapid, not labor intensive, and highly selective for the separation of hemoglobin variants. Combination of both methods with some conventional methods, such as isoelectrofocusing, allows identification of Hbs C, E, O-Arab, S, and D-Punjab, as well as their quantification. We have demonstrated that the conventional electrophoresis methods (electrophoresis at pH 6.5 in citrate agar gel and electrophoresis at pH 8.6 on cellulose acetate) can be advantageously replaced by the present capillary zone electrophoresis methods in a clinical laboratory practice for the detection and quantification of hemoglobin variants.

Electrophoresis, Capillary↗
PubMed1999

[Rapid detection of trisomy 21 by quantitative polymerase chain reaction].

OBJECTIVE: Studying gene diagnosis of trisomy 21 by means of quantitative polymerase chain reaction(PCR). METHODS: Polymorphic short tandem repeat(STR) at D21S11 served as the gene marker, and a pair of primers was synthesized. The samples from 11 normal subjects and 28 cases of trisomy 21 were analyzed by quantitative PCR. RESULTS: 10 normal subjects showed two bands with a ratio of 1:1, and a subject showed one band. Of 28 cases of trisomy 21, 24 cases showed two bands with a ratio of 2:1, 3 cases showed three bands with a ratio of 1:1:1, and 1 case showed one band. CONCLUSION: Polymorphic STR at D21S11 is a valuable gene marker for diagnosis of trisomy 21. Trisomy 21 can be diagnosed rapidly and accurately within 24 hours by quantitative PCR.

Down Syndrome↗
PubMed1999

Effects of the configuration of a multi-epitope chimeric malaria DNA vaccine on its antigenicity to mice.

OBJECTIVE: Four B and Th cell epitopes were selected from conservative domain of Plasmodium falciparum antigens to construct two groups of chimeric malaria DNA vaccines with different configurations and their antigenicities were studied. METHODS: The partially synthesized oligonucleotide was annealed, PCR amplified and cloned into a mammalian cell expression vector. By using a pair of isocaudamers on the vector, different single copies of B epitopes were multiplied and were tenderly stringed into two groups of chimeric DNA vaccine with different configurations. BALB/c mice were immunized with these DNA plasmids by either intramuscular or intradermal injections. RESULTS: The antisera from the immunized mice tested by ELISA showed that only the configuration which had a single copy of universal T helper cell epitope, CS. T3, located at the C terminal of the multi-copy B cell epitopes induced a high antibody response. The T helper cell epitope at any other position of the peptide, or the double T helper cell epitopes configured with the B cell epitopes did not enhance antibody response, and some configurations even decreased the humoral response to a B cell epitope. CONCLUSION: This study demonstrated that both combination and configuration of the epitope may affect the antigenicity of a chimeric multiple antigen.

Animals↗
PubMed1999

Lack of human papillomavirus types 16 and 18 DNA in esophageal squamous carcinoma cell lines.

OBJECTIVE: To assess the role of human papillomavirus (HPV) in the carcinogenesis of the esophageal carcinomas in a high-incidence geographical area in Northern China. METHODS: The presence of HPV types 16 and 18 DNA in the three esophageal carcinoma cell lines was examined by type-specific polymerase chain reaction (PCR). RESULTS: No HPV type 16 and 18 DNA was found in any of the three esophageal carcinoma cell lines. CONCLUSION: The results suggest that the role of HPV in human esophageal carcinogenesis in a high-incidence geographical area remains to be elucidated.

Adult↗
PubMed1999

[Adenovirus-transferred antisense c-myc selectively induces tumor cell cycle arrest and apoptosis].

OBJECTIVES: To construct the recombinant adenovirus encoding antisense c-myc fragment and to investigate its effect on tumor cell proliferation and apoptosis. METHODS: The shuttle plasmid encoding antisense c-myc was constructed by cloning c-myc cDNA fragment in the reverse direction into the pAdCMV. Then the plasmid pJM17 and the shuttle plasmid were co-transferred into 293 cells with liposome for homologous recombination to acquire recombinant adenovirus. The cell cycle, apoptosis, and the expression of related genes of the in vitro transferred human adenocarcinoma cell lines GLC-82 and SPC-A-1, as well as the human embryonic diploid lung cell line 2BS were studied. The effects on colony formation and treating effect on transplanted tumor in nude mice were also studied. RESULTS: The recombinant adenovirus encoding antisense c-myc fragment was obtained with the titer of 3.3 x 10(10) pfu/ml. RT-PCR and Western blot showed that the expression of c-myc was reduced 3-5 days after infection. Simultaneously, obvious G1 arrest and apoptosis as well as the alteration of cyclin D1, bcl-2 and bax gene expression were observed in the infected tumor cells. The infected 2BS cells showed no such changes. The colony formation ability in vitro and the growth of the hetero-transplanted tumor in nude mice were also reduced. CONCLUSIONS: The reduced expression of c-myc gene could induce human adenocarcinoma cells tumor-cell-specific G1 arrest and apoptosis, while it has no effect on normal cell.

Adenocarcinoma↗
PubMed1999

[In vitro and in vivo evaluation of the safety of Ad-RA538].

OBJECTIVE: To assess the safety of adenovirus-mediated transfer of the RA538 (Ad-RA538) for the treatment of cancer and to furthermore in preparation for a clinical trial of Ad-RA538. METHODS: RT-PCR was used to detect the transcription of Ad-RA538 in HeLa cells infected with extracts from HeLa cells previously infected with Ad5-RA538. Cell counting was made to observe the effects of Ad-RA538 on the growth of the normal human fetal lung cell line 2BS. The virus was intraperitoneally injected into 2 groups of BalB/C mice at a dosage of 10(7) pfu and 10(9) pfu. Blood samples were taken from the mice to test the liver and renal function. PCR were used to screen the vital organs for the presence of adenovirus DNA. Microscopic examination of the vital organs was performed to observe the pathogenicity of Ad-RA538. RESULTS: Ad-RA538 was a replication-defective virus. It could infect 2BS cells effectively, but could not inhibit 2BS cell growth. No mouse died and no signs of general toxicity were seen following intraperitoneal injection of Ad-RA538. The adenoviral vector was present in the liver, spleen, kidney and stomach of mice injected with 10(9) pfu Ad-RA538. Six and 12 days after injection, mild inflammation was observed in the liver of mice received 10(9) pfu Ad-RA538. CONCLUSION: Ad-RA538 is safe both in vivo and in vitro, and clinical trials of Ad-RA538 can be performed.

Adenovirus E1 Proteins↗
PubMed1999

[Apoptosis of human melanoma cell line WM-983A by p16 gene transduction].

OBJECTIVE: To further understand the mechanism of action of the tumor-suppressor gene p16. METHODS: An adenoviral expression vector with full length cDNA of p16 gene insert was constructed (Ad-p16) and transfected into WM-983A cells, the p16 gene of which was point mutated at codon 126. The effect of exogenous p16 gene on the growth of WM-983A cells was examined in vitro and in vivo. RESULTS: Expression of p16 gene in WM-983A cells was confirmed by Western blot. The in vitro growth of the Ad-p16 transfected WM-983A cells was significantly inhibited (inhibition rate: 78%) as compared to mock (Ad-LacZ) transfected WM-983A cells. Colony-forming activity in vitro of the Ad-p16 transfected WM-983A cells was completely inhibited. Morphologically, the Ad-p16 transfected cells appeared apoptotic which was confirmed by the appearance of pre-G1 on flow cytometry and DNA fragmentation. The growth of WM-983A xenografts in nude mice was retarded by intra-tumoral injection of Ad-p16. CONCLUSION: p16 gene participates in the induction of cell apoptosis. It is promising to use it for gene therapy of cancer, especially when combined with other apopptosis-inducing agents.

Animals↗
PubMed1999

[Taxonomic studies on Echinochasmus fujienensis and its related species by random amplified polymorphic DNA analysis and experimental infection].

AIM: To explore the identification and differentiation of Echinochasmus fujianensis, Echinochasmus japonicus(Jiangxi strain and Fujian strain), Echinochasmus liliputanus Anhui and Echinochasmus per foliatus Hubei. METHODS: Random amplified polymorphic DNA analysis (RAPD) and experimental animal infection were performed. RESULTS: 469 polymorphic DNA fragments were obtained by 28 primers from 4 Echinochasmus species and strains in Fujian, Anhui and Jiangxi. 20.8% and 97.6% of the fragments in Echinochasmus fujianensis were the same as those in Echinochasmus japonicus Fujian strain and in Echinochasmus liliputanus Anhui, respectively, 99.7% of the fragments were the same between Echinochasmus japonicus Jiangxi strain and Fujian strain. CONCLUSION: Echinochasmus fujianensis and Echinochasmus liliputanus Anhui are the same species. Echinochasmus fujienensis is an independent species different from Echinochasmus japonicus. Polyinfection of Echinochasmus fujianensis, Echinochasmus japonicus and Echinochasmus perfoliatus exist in all the 3 provinces, Hubei, Anbui and Fujian, of which Echinochasmus fujienensis is a dominant species.

Animals↗
PubMed1999

Evaluation of a capillary electrophoresis method for routine determination of hemoglobins A2 and F.

Hemoglobin A2 (Hb A2) and hemoglobin F (Hb F) are important analytes in the diagnosis and follow up of Hb diseases. We evaluated a new capillary zone electrophoresis (CZE) kit for Hb A2 and Hb F measurements. The imprecision ranged from 3% to 6% for Hb A2 and Hb F at physiological and pathological concentrations. The method compared well with cation-exchange HPLC for Hb A2 and Hb F and with anion-exchange chromatography in microcolumns (MAEC), for Hb A2. Nevertheless, higher results were obtained [Hb A2 CZE (%) = 1.233 Hb A2 HPLC - 0.2; Hb A2 CZE (%) = 1.190 Hb A2 MAEC + 0.1; Hb FCZE (%) = 1.118 Hb FHPLC + 0.4], and new reference values had to be determined (Hb A2 2.7-3.8%; Hb F <1.2%). Quantification of Hb A2 was not influenced by Hb S. Measurement of Hb F was accurate and precise except at low concentrations in Hb AS patients. This new CZE kit is rapid, precise, and reliable, and seems appropriate for use in clinical laboratories.

Electrophoresis, Capillary↗
PubMed1998

Transgenic knockout mice exclusively expressing human hemoglobin S after transfer of a 240-kb betas-globin yeast artificial chromosome: A mouse model of sickle cell anemia.

Sickle cell anemia (SCA) and thalassemia are among the most common genetic diseases worldwide. Current approaches to the development of murine models of SCA involve the elimination of functional murine alpha- and beta-globin genes and substitution with human alpha and betas transgenes. Recently, two groups have produced mice that exclusively express human HbS. The transgenic lines used in these studies were produced by coinjection of human alpha-, gamma-, and beta-globin constructs. Thus, all of the transgenes are integrated at a single chromosomal site. Studies in transgenic mice have demonstrated that the normal gene order and spatial organization of the members of the human beta-globin gene family are required for appropriate developmental and stage-restricted expression of the genes. As the cis-acting sequences that participate in activation and silencing of the gamma- and beta-globin genes are not fully defined, murine models that preserve the normal structure of the locus are likely to have significant advantages for validating future therapies for SCA. To produce a model of SCA that recapitulates not only the phenotype, but also the genotype of patients with SCA, we have generated mice that exclusively express HbS after transfer of a 240-kb betas yeast artificial chromosome. These mice have hemolytic anemia, 10% irreversibly sickled cells in their peripheral blood, reticulocytosis, and other phenotypic features of SCA.

Anemia, Sickle Cell↗
PubMed1998

Four novel mutations of the connexin 32 gene in four Japanese families with Charcot-Marie-Tooth disease type 1.

DNA-based mutation analysis on the connexin 32 gene was performed in 49 families with Charcot-Marie-Tooth disease (CMT) type 1 but without duplication involving the chromosomal region, 17p12-p11.2. Mutations were identified in five of the 49 families, and four of the five mutations were hitherto undescribed: Va137Met, Glu57His, Arg142Glu, Val177Ala. X-linked CMT sometimes lacks evidence for X-linked transmission and cannot be differentiated from CMT type 2, especially in females with mildly decreased nerve conduction velocity. Therefore, molecular analysis is useful for molecular pathology of their disease.

Adolescent↗
PubMed1998

Separation of human globin chains by micellar electrokinetic capillary chromatography.

A new separation method of human globin chains by micellar electrokinetic capillary chromatography (MECC) is described. In this method, a 25 mM phosphate buffer (pH 2.5) containing 7 M urea and 1% (w/v) reduced Triton X-100 buffer system was used. All experiments were performed in a 47 cmx50 microm I.D. uncoated fused-silica capillary. The separation voltage was set at 19 kV. Normal globin chains derived from normal adults and newborns, alpha, beta, delta, Ggamma and Agamma globin chains as well as common variant globin chains were successfully separated within 20 min. High reproducible migration times of globin chains (CVs of intra- and inter-assay were less than 1% and 2% respectively), and quantification of Ggamma and Agamma chains (CVs for intra- and inter-assay were less than 5% and 10%, respectively) were obtained. This new MECC method provides primary information on structural modification of globin chains. It can be an important diagnostic tool in clinical laboratory practice in the field of hemoglobinopathies.

Adult↗
PubMed1998

Mutagenesis associated with nitric oxide production in macrophages.

To better understand the mechanisms through which persistent infections/inflammation increase cancer risks, we assessed the potential genotoxic properties of NO produced by macrophages. We recently showed that mouse macrophage RAW264.7 cells were capable of resuming exponential growth after stimulation for NO production by interferon-gamma (IFN-gamma) and/or lipopolysaccharide. Here, we report that increases in mutant fraction (MF) in the endogenous, X-linked, hprt gene of the cells are associated with NO exposure. Cells stimulated with 100 units/ml IFN-gamma continuously for 14 and 23 days produced a total of 9.8 and 14 micromol of NO per 10(7) cells, respectively. MFs in the hprt gene of NO-producing cells were 16.6 and 31.3 x 10(-5), respectively, compared with 2.2 and 2.5 x 10(-5) in untreated cells. Addition of an NO synthase inhibitor, N-monomethyl-L-arginine, to the culture medium decreased NO production and MF by 90% and 85%, respectively. Reverse transcription-PCR and DNA sequencing revealed that NO-associated hprt mutations did not differ significantly from those arising spontaneously, with the exception that certain small deletions/insertions and multiple exon deletions were observed only in the former. MF also increased significantly in cells stimulated for only 4 days with lipopolysaccharide plus IFN-gamma for higher rates of NO production. The types and proportion of hprt mutations induced under these conditions were strikingly similar to those associated with long-term NO exposure. These results indicate that NO exposure results in gene mutations in RAW264.7 cells through mechanisms yet to be identified and may also contribute to spontaneous mutagenesis.

Animals↗
PubMed1998

Potent, selective, and orally bioavailable tricyclic pyridyl acetamide N-oxide inhibitors of farnesyl protein transferase with enhanced in vivo antitumor activity.

We previously reported compound 1 as a potent farnesyl protein transferase (FPT) inhibitor that exhibited reasonable pharmacokinetic stability and showed moderate in vivo activity against a variety of tumor cell lines. The analogous C-11 single compound, pyridylacetamide 2, was found to be more potent than 1 in FPT inhibition. Further studies showed that modification of the ethano bridge of the tricyclic ring system by conversion into a double bond with concomitant introduction of a single bond at C-11 piperidine resulted in compound 3 that had superior FPT activity and pharmacokinetic stability. Compound 4, a 5-bromo-substituted analogue of 3, showed improved FPT activity, had good cellular activity, and demonstrated a remarkably improved pharmacokinetic profile with AUC of 84.9 and t1/2 of 82 min. Compound4 inhibited the growth of solid tumor in DLD-1 model by 70% at 50 mpk and 52% at 10 mpk.

Administration, Oral↗
PubMed1998

Enhancement of blue-light sensitivity of Arabidopsis seedlings by a blue light receptor cryptochrome 2.

Cryptochrome is a group of flavin-type blue light receptors that regulate plant growth and development. The function of Arabidopsis cryptochrome 2 in the early photomorphogenesis of seedlings was studied by using transgenic plants overexpressing CRY2 protein, and cry2 mutant plants accumulating no CRY2 protein. It is found that cryptochrome 2 mediates blue light-dependent inhibition of hypocotyl elongation and stimulation of cotyledon opening under low intensities of blue light. In contrast to CRY1, the expression of CRY2 is rapidly down-regulated by blue light in a light-intensity dependent manner, which provides a molecular mechanism to explain at least in part that cryptochrome 2 functions primarily under low light during the early development of seedlings.

Amino Acid Sequence↗