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Biomedical subjects

J Miao

Publications and source records attributed to J Miao.

At least 55 records · Page 3Linked to original sources

Progressive myoclonus epilepsy EPM1 locus maps to a 175-kb interval in distal 21q.

The EPM1 locus responsible for progressive myoclonus epilepsy of Unverricht-Lundborg type (MIM 254800) maps to a region in distal chromosome 21q where positional cloning has been hampered by the lack of physical and genetic mapping resolution. We here report the use of a recently constituted contig of cosmid, BAC, and P1 clones that allowed new polymorphic markers to be positioned. These were typed in 53 unrelated disease families from an isolated Finnish population in which a putative single ancestral EPM1 mutation has segregated for an estimated 100 generations. By thus exploiting historical recombinations in haplotype analysis, EPM1 could be assigned to the approximately 175-kb interval between the markers D21S2040 and D21S1259.

Adult↗

[Clinical pathological study on 23 pulmonary embolism proved by autopsy].

OBJECTIVE: To improve the early diagnosis of pulmonary embolism. METHODS: The clinical and pathological data of 23 cases of autopsy-proved pulmonary embolism in Beijing hospital from 1951 to 1994 were reviewed. RESULTS: 23 patients: male 18, female 5, age ranged from 34 to 82 years. Misdiagnosis rate was 87%. The unexplained dyspnea, chest pain, sudden onset dyspnea and sudden death were the most frequent symptoms. Arterial blood gas data frequently demonstrated hypoxia and hypocapnia. The most important causes of an incorrect diagnosis could be the failure of suspecting pulmonary embolism, because of the variability of clinical manifestation and the presence of other concomitant diseases. CONCLUSION: All patients with otherwise severe unexplained dyspnea or chest pain have to be sent for perfusion lung scintigraphy, accordingly, underdetection of pulmonary embolism and mortality of pulmonary embolism could be reduced.

Adult↗

Expression of human interleukin-11 cDNA in E. coli.

A 551-bp hIL-11 gene fragment that includes no nucleotide sequences encoding signal polypeptide and the initial 8 amino acids of the mature protein was cloned into a high-level expression vector pEx31B of E. coli. The authors identified the recombinant plasmid, designated pEx31-IL11, by restriction endonucleases digestion and DNA sequencing. The resulting recombinant plasmids were then used to transform E. coli strain HB101, and expression in the PL promoter system, which is temperature-regulated, was achieved. The expressed fusion protein amounts to 50% of total bacterial proteins. The hIL-11 protein expressed in E. coli was fused to the N-terminal 99 amino acids of the MS2 polymerase to form the inclusion body. These recombinant proteins can be purified to about 80% by extracting inclusion body with urea. One IL-6-dependent cell line 7 TD1 was used for bioassay. The recombinant hIL-11 protein was preliminarily purified and renatured to a specific activity of 10(5)U/mg, even in the presence of an excess of a neutralizing anti-IL-6 antibody.

DNA, Complementary↗

[A randomized controlled clinical trial of sulperazone as compared with cefotaxime in the treatment of bacterial infections].

A randomized controlled clinical trial of sulperazone as compared with cefotaxime was conducted. 207 patients with bacterial infections entered the study. The overall clinical efficacy rate of sulperazone and cefotaxime was 95.15% and 90.38% respectively. The bacterial clearance rate in the two groups was 84.7% and 80.6% respectively. The adverse drug reactions in both groups were mild with the rate of 7.77% and 8.65% respectively; there was no statistical difference between the two groups. The results of disc susceptibility test showed that the sensitive rate of the clinical isolates to sulperazone was 90.9%, being significantly higher than that of cefotaxime 69.3% (P < 0.001).

Adolescent↗

Cloning, expression and purification of the ligand-binding region of human IL-6R in E. coli and its preliminary functional identification.

The ligand-binding region of human IL-6R is taken as the target gene fragment to be cloned and expressed. With pET-3b as expressing vector, two recombinants pET-6R(B) and pET-6R(B)4 have been constructed encoding the ligand-binding region (28 kD) of hIL-6R and its dimmer (53 kD), respectively. After induction with IPTG, they produced two proteins rIL6R-28 of 28 kD and rIL6R-53 of 53 kD amounting to 50% and 30% of total bacteria proteins, respectively. The expressed products were mainly recovered as inclusion bodies. After purification and renaturation, both of them were capable of augmenting the growth-stimulating effect of IL-6 on 7TD1 cells, an IL-6 dependent cell line. The result of ELISA also revealed that both rIL6R-28 and rIL6R-53 had the obvious ligand-binding activity.

Antigens, CD↗

[The clinical significance of Pseudomonas aeruginosa typing and R-plasmid and DNA molecule hybridization in patients with cor pulmonale].

600 strains of Pa were typed from the sputa of patients with cor pulmonale in 8 hospitals. P6 type stoods first, accounting for 25.6%, and the types of PA and the difference in distribution of Pseudomonas from different districts and hospitals, might be used as an important parameter in the epidemiological study. 68 of 150 strains from patients with cor pulmonale contained plasmids. 60 isolates harboring plasmids colonies were hybridized with 6 beta-lactamase and 6 aminoglycoside modifying enzyme resistance gene probes. The experiments demonstrated the 3 strains isolated from patients with nosocomial infection came from one same strain and got a new resistance gene during spreading of nosocomial infection, and these isolates contained homologous sequence with OXA-2 and AAC (6')-1b probes.

DNA Fingerprinting↗

HIV-1 infection in pigtailed macaques.

Four pigtailed macaques were inoculated with autologous cells expressing low levels of human immunodeficiency virus type 1 (HIV-1). During the first 10 weeks, infectious virus was recovered from peripheral blood mononuclear cells (PBMCs) and lymph nodes from three of the animals. Subsequently, HIV-1 DNA was frequently detected in uncultured PBMCs from all three animals, and virus was isolated from one of them at weeks 38 and 61. The fourth animal, which was rechallenged at week 10 with cell-free virus isolated from one of the others, never became virus isolation positive, but harbored HIV-1 proviral genomes. These virus infections were accompanied by the development of varied HIV-1-specific humoral immune responses. Antibodies to gp160 were first apparent at week 8 in the three initially infected animals and persisted. The animal from whom virus was isolated at late times also developed persisting antibodies to HIV-1 p24 and gp120. Antibodies to gp120 and gp160 became apparent in the rechallenged animal at 1 week following reinoculation, but they waned with time. In vivo passage of the virus was attempted at week 6. One recipient pigtailed macaque and one recipient cynomolgus monkey failed to become detectably infected following transfusion of virus-positive blood and lymph node cells. The long-term presence of HIV-1-specific antibodies and proviral genomes in these animals, and the recovery of infectious virus more than 1 year following inoculation, are indicative of persistent infection, and confirm previous reports that pigtailed macaques are susceptible to HIV-1.

Animals↗

Localization of a gene for progressive myoclonus epilepsy to chromosome 21q22.

Progressive myoclonus epilepsy of Univerricht-Lundborg type is a clinically defined entity among the progressive myoclonus epilepsies. It is an autosomal recessive disorder. The underlying biochemical defect is unknown. We used linkage analysis to localize the gene in 12 families with the aid of polymorphic DNA markers. Close linkage was detected with three markers on distal chromosome 21. The loci BCEI and D21S154 gave the highest positive logarithm-of-odds (lod) scores of 5.49 and 4.25, respectively, at zero recombination. The third locus, D21S112, gave a lod score of 6.91 at a recombination fraction of 0.034. There was no evidence of heterogeneity. Multipoint lod scores calculated against a fixed map of the three marker loci gave a maximum four-point lod score of 10.08 at a location of the disease gene at 6.0 centimorgans distal to locus BCEI and 0.8 centimorgan proximal to locus D21S154. As markers BCEI and D21S154 have previously been localized to 21q22.3 by physical methods, our findings place the EMP1 gene locus (for progressive myoclonus epilepsy of the Unverricht-Lundborg type) in chromosome 21 band q22.3. This finding provides an opportunity to test several other epilepsy phenotypes, particularly the so-called Ramsay Hunt syndrome, for linkage to the same locus. It also is a starting point toward isolating and characterizing the gene and its protein product.

Chromosome Mapping↗

Mutagenicity of Alternaria alternata and Penicillium cyclopium isolated from grains in an area of high incidence of oesophageal cancer--Linxian, China.

In order to study the relationship between daily consumption of mouldy food and the incidence of oesophageal cancer, we examined the mutagenicity of Alternaria alternata and Penicillium cyclopium, which seriously contaminate grain in Linxian county, China. We first examined extracts of cultured strains of A. alternata. In the reverse mutation test, positive results were obtained in 85% of strains; positive results were seen in 84% of 19/20 strains in the rec assay. Eight of ten strains induced sister chromatid exchange, and two of eight strains induced chromatid breaks. Six of seven strains induced unscheduled DNA synthesis and DNA synthesis inhibition. One extract induced a higher frequency of sister chromatid exchange in lymphocytes from normal persons than in those from patients with oesophageal cancer, and the spontaneous break-points in patients were related to fragile sites and neoplasia-associated break-points. The toxins alternariol and its monomethyl ether, produced by A. alternata, were examined in the reverse mutation assay and for unscheduled DNA synthesis. The results were similar to those obtained with extracts of the different strains. Alternariol had a four to eight times greater effect than its monomethyl ether. Of 24 strains of P. cyclopium isolated from cereals in Linxian, four were cultured with rice and 19 in Raulin-Thom medium. Cultures in Raulin-Thorn medium, solution and hyphae were then extracted. The strains cultured with rice induced sister chromatid exchange, unscheduled DNA synthesis and DNA synthesis inhibition. The solution extracts of 14 strains were positive in the rec assay, and five strains were positive in the reverse mutation test.(ABSTRACT TRUNCATED AT 250 WORDS)

Alternaria↗

Relationships between Alternaria alternata and oesophageal cancer.

Although it is uncertain whether fungi can produce the mycotoxins that induce oesophageal cancer, the tumorigenicity of fungi isolated from grain in an area of high incidence of oesophageal cancer has been confirmed. Alternaria alternata is a fungus of importance in this respect. The contamination rate with A. alternata in corn from areas of high morbidity from oesophageal cancer is higher than that in low-morbidity areas. Extracts of A. alternata induced reverse mutation in Escherichia coli, unscheduled DNA synthesis in cultured human amnion FL cells, chromosomal aberrations and sister chromatid exchange in human peripheral blood lymphocytes, mutation in V79 cells and transformation of NIH3T3 cells. Alternariol methyl ether, which is an active compound produced by this fungus, also induced transformation of NIH3T3 cells, and the transformed cells grew in soft agar and were tumorigenic in nude mice. Food mildewed by this fungus induced forestomach tumours in rats. Thus, (i) A. alternata is tumorigenic, (ii) it has been isolated from corn in an area of high incidence of oesophageal cancer, and (iii) contamination with A. alternata in the area of high incidence is higher than that in the area of low incidence. We therefore believe that A. alternata is one of the causes of human oesophageal cancer.

Alternaria↗

X linked neonatal myotubular myopathy: one recombination detected with four polymorphic DNA markers from Xq28.

A three generation family with X linked myotubular myopathy (MTM1) was studied with several polymorphic markers from the distal long arm of the X chromosome. A recombination between the disease gene and four markers (loci DXS52, DXS134, DXS15, F8C) from the Xq28 cluster was detected. A new polymorphic marker (U6.2) defining the locus DXS304 in the Xq27-28 region proximal to the Xq28 cluster did not show any recombination with MTM1. These results suggest the following order of loci in distal Xq: cen-DXS42-DXS105-(DXS304, MTM1)-(DXS52, DXS134, DXS15, F8C)-tel.

Cell Nucleus↗

[Amplification and rearrangement of proto-oncogene c-abl in human leukemia cells].

DNA of peripheral blood or bone marrow leukocytes from 8 normal subjects, 7 cases of acute lymphocytic leukemia (ALL), 2 of acute myelogenous leukemia (AML) and 1 of chronic myelogenous leukemia (CML), having been digested by endonuclease Eco RI or Pst I separately, was hybridized with the probes of 3' fragment (Pst I/Hind III) or 5' fragment (Hinc II/Pst I) of Abelson murine leukemia virus (A-MuLV) oncogene v-abl. The proto-oncogene c-abl, which is homologous to v-abl, was found amplified in 4 ALL, 1 CML and 1 AML. In one of these 4 ALL, c-abl was amplified even over 100 times. A new c-abl BamH I fragment with 6.7 kilobase pairs (kb) in length was observed in 2 ALL and 1 CML out of these 6 cases with amplification, but none of this fragment was found in the normal subjects or other leukemia patients. These 3 patients with the presence of 6.7 kb fragment were high risk ones and 2 of them had died, suggesting that 6.7 kb fragment be the index of poor prognosis. The amplification and rearrangement of c-abl imply the activation of proto-oncogene in leukemogenesis.

Gene Amplification↗

Activated c-Ha-ras oncogene with a guanine to thymine transversion at the twelfth codon in a human stomach cancer cell line.

The rat fibroblast cell line Rat 1 was transfected with total DNA of a gastrocarcinoma cell line, BGC-823. The transforming gene was cloned from the genomic library of the secondary transformants using in situ hybridization with a probe of the human Alu repeat sequence. This cloned gene is homologous to the protooncogene c-Ha-ras. The activation lesion of the transforming gene was identified by sequence analysis as a single nucleotide substitution of thymine for guanine in the 12th codon. This results in the substitution of valine for glycine at the 12th amino acid of the Mr 21,000 protein.

Adenocarcinoma↗

Molecular cloning and nucleotide sequence analysis of a transforming gene from human gastrocarcinoma cell line.

Mouse and rat fibroblasts were transfected with total DNA from human gastrocarcinoma cell line BGC-823. It was shown by hybridization assay that the genome of one of the rat secondary foci contains transforming genes from the human gastrocarcinoma cell line, which are homologous to the protooncogene c-Ha-ras in the normal cells. The genomic library of the rat secondary foci was constructed, using lambda phage EMBL3 as the vector. The transforming gene Ha-ras of the human gastrocarcinoma cell was thus cloned by screening the library with the probes of human Alu repeat sequence and c-Ha-ras. The nucleotide sequences of the first and second exons were analysed by M13-dideoxy method. The result shows that the nucleotide sequence of the transforming gene is the same as that of the normal protooncogene except one nucleotide difference in the first exon.

Amino Acid Sequence↗

[DNA transformation activity of a human gastrocarcinoma cell line].

DNAs of three cell lines of human gastrocarcinoma (MGC-803, BGC-823 and PACM-82) and two fresh solid tumors of human stomach cancer were used to transfect NIH3T3 and Rat-1 cells. The transformed cells were selected with high concentration of glucose and low concentration of serum, or with medium containing Geneticin (G418) after co-transfection of pSVneo and DNAs of stomach cancer cell line or primary transformants. From the second round transfection, we had obtained transformants which could grow with high colony forming efficiency in soft agarose and were tumorigenic in nude mice. The southern blot analysis showed that the cellular DNA of the transformants contained human Alu repeat sequence and the transformed gene from stomach cancer cell line (BGC-823) and was homologous to proto-oncogene c-Ha-ras. The transforming gene is able to induce neoplastic transformation of NIH3T3 and Rat-1 cells.

Animals↗

Spatial distribution of [14C]-lidocaine and blood flow in transmural and lateral border zones of ischemic canine myocardium.

The purpose of this study was to determine the spatial distribution of lidocaine relative to blood flow in ischemic, normal and border zone canine myocardium. Ischemic zone tissue was distinguished from normal zone tissue by a special microsphere technique in adjacent sections 4 to 5 mm wide from the center to the lateral border of the ischemic region in 14 open chest dogs. Gamma-labeled microspheres were separated by a special technique from carbon-14 ([14C])-lidocaine in the same tissue sample. Blood flow (mean value +/- 1 standard deviation) was reduced to 46 +/- 25 percent of normal in the ischemic subepicardium and 17 +/- 18 percent of normal in the subendocardium. [14C]-lidocaine was 0.56 +/- 0.12 microgram/g in normal myocardium 10 minutes after bolus injection of [14C]-lidocaine; it was reduced to 91 +/- 15 percent of normal in ischemic subepicardium and 58 +/- 12 percent of normal in the subendocardium. Blood flow and lidocaine concentration were uniformly lowest in gross samples from the central and intermediate ischemic zones, and highest in the gross samples from the border normal zone (p less than 0.05). The values for flow and lidocaine in samples from the border ischemic zone were intermediate, that is, higher than values from central ischemic (p less than 0.05) and lower than values from border normal zone samples (p less than 0.05). However, the labeling technique for normal zone tissue revealed that the values of blood flow and lidocaine in the gross samples from the lateral border of the ischemic zone were intermediate between those of adjacent ischemic and normal samples because of the mixture of overlapping normal and ischemic tissues components--not because of a unique mildly ischemic region. Both blood flow and lidocaine concentration were lower in the subendocardial third than in the subepicardial third of the ischemic zone (p less than 0.05) even after the contribution of normal zone tissue was subtracted, suggesting a gradient of ischemia across the transmural border zone. In conclusion, lidocaine is distributed uniformly in ischemic components from the center to the lateral border of the ischemic zone, but there is an endocardial to epicardial gradient. Both lateral and transmural border zone distributions must be considered to understand the mechanisms of drug effects in myocardial ischemia.

Animals↗

Diet and vitamin nutrition of the high esophageal cancer risk population in Linxian, China.

In order to examine the nutritional status of the population of Linxian (in Henan, China) known to be at high risk for esophageal cancer (EC), we analyzed blood samples and conducted 3-day dietary surveys on 3 groups of normal adults, age 40-50, from Henan province. Two groups were from Linxian, where the EC mortality rate is about 138/100,000 (Group C, consisting of volunteers with a family history of EC, and Group NC, consisting of those who did not have a family history of EC). The third group, Group F, was from Fanxian, where the EC mortality rate is about 24/100,000. The average plasma retinol level was found to be about 0.27 micrograms/ml in all 3 groups; about 13%-20% of the individuals had a level less than 0.2 micrograms/ml. The average plasma carotene level of all 3 groups appeared to be adequate, ranging from 0.5-1.0 micrograms/ml. About 23% of the subjects in both Groups C and NC had low (less than 2 micrograms/ml) plasma ascorbate levels; in Group F, 16% had low ascorbate levels. Less than 12% of the individuals in each of the 3 groups had erythrocyte glutathione reductase activation coefficients less than 1.2, indicating a widely occurring riboflavin deficiency. Consumption of alcoholic beverages was not found to be a contributing factor to EC. The results of the present study reveal a deficiency of vitamins in the study populations, but do not indicate that the average nutritional status of the volunteers in Group C was worse than that of Groups NC and F with regard to vitamin A, ascorbate, and riboflavin.

Adult↗