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Biomedical subjects

J G Chang

Publications and source records attributed to J G Chang.

At least 145 records · Page 8Linked to original sources

ABO genotyping by polymerase chain reaction.

ABO blood group system's genotyping by polymerase chain reaction in genomic DNA level is developed. The positions of nucleotide 258 and 700 of cDNA from A transferase were used to distinguish A, B, and O alleles by restriction enzyme digestion. To identify the 258th nucleotide, a 199- or 200-bp DNA fragment was amplified by PCR and digested with Kpn I. For the 700th nucleotide, a 128-bp PCR amplified fragment was designed and digested with Alu I. By examining the DNA fragment digested patterns, ABO genotypes were easily determined. Results obtained using this method on 20 ABO-known peripheral blood samples showed that this new technique could provide accurate ABO genotype. Biologic forensic samples, such as, blood stains, saliva stains, semen stains, hair, bone tissue, and semen contaminated with vaginal secretion were also successfully typed. This rapid, sensitive and reliable method should be applicable not only in forensic identification but also in medical examination.

ABO Blood-Group System↗

Rapid prenatal diagnosis of Hb Bart's hydrops fetalis in southeast Asia area by polymerase chain reaction.

We used the polymerase chain reaction (PCR) to amplify the breakpoint area of alpha-thalassemia-1 of Southeast Asia type and several parts of the alpha-globin gene cluster to make a differential diagnosis between alpha-thalassemia-1 and Hb Bart's hydrops fetalis. The procedure involved three primers to detect the homozygote of alpha-thalassemia-1, then amplifies the other alpha-globin gene cluster with three other pairs of primers to double check the results. The PCR products were checked again by allele specific probes. Twenty-two cases were diagnosed prenatally, two were normal, 17 were alpha-thalassemia-1, and three Hb Bart's hydrops fetalis. All cases were confirmed either by Southern blot hybridization or follow-up by sonography or after delivery. No false positive or false negative results were obtained by our strigent procedure. We conclude it to be a rapid, accurate and economic method.

Asia, Southeastern↗

Detection of beta-globin gene mutations by polymerase chain reaction.

The polymerase chain reaction and oligonucleotide probe hybridization technique were applied to the detection of two common mutations of the beta-globin gene found in Chinese, namely the 4-base pair deletion at the 41-42 codons and the C to T substitution at nucleotide 654 of IVS-2. The accuracy of the method was established using beta-thalassemia cases with known mutations or haplotypes of the restriction fragment length polymorphism (RFLP). A further 11 cases of thalassemia intermediate and thalassemia minor were then analysed with the same approach. Our results showed that 5 of the 11 cases carried the TCTT-deletion at codons 41-42. Our method is economical both in terms of materials and time needed and in an alternative to the use of the molecular RFLP approach in the prenatal diagnosis of beta-thalassemia.

Base Sequence↗

Mutation analysis of the ras gene in myelocytic leukemia by polymerase chain reaction and oligonucleotide probes.

Twenty-nine patients with acute myelocytic leukemia (AML) and 14 patients with Philadelphia chromosome-positive chronic myelocytic leukemia (CML) were analyzed to detect the presence of mutations in their ras genes by the polymerase chain reaction and oligonucleotide hybridization methods. Deoxyribonucleic acid (DNA) isolated from blood or bone marrow samples was screened for mutations in codons 12, 13 and 61 of N-ras and in codons 12 and 61 of K-ras and H-ras. We detected mutations of the ras gene in 7 patients with AML (7/29), all in N-ras. The mutations were 3 GGT- greater than GAT transitions in codon 12, 1 GGT- greater than TGT transition in codon 13, and 3 CAA- greater than AAA transitions in codon 61. No correlation has been observed between French-American-British subtypes and the incidence of N-ras mutation, nor between cytogenetic changes and the incidence of N-ras mutation. All ras gene mutations detected by the oligonucleotide hybridization method were further confirmed by direct sequencing. No mutations were detected in ras genes in samples from the 14 Philadelphia chromosome-positive CML patients (12 in chronic phase, 2 in blastic phase). These findings are in line with previous results indicating that ras gene mutations in the codons tested play only a small role in the tumorigenesis of CML.

Adolescent↗

Analysis of ras gene mutations in gastrointestinal cancers.

Point mutations of c-Ki-ras genes were analyzed in 33 samples of human gastrointestinal malignancy. DNA extracted from the frozen tissue was amplified by polymerase chain reaction (PCR) and analyzed by direct sequencing and slot-blot hybridization at codons 12, 13 and 61 of c-Ki-ras. In 7 cases out of 19 colorectal cancers, point mutations were found: 3 cases at codon 12, 1 at codon 13, 2 at codon 61 of c-Ki-ras and one case had double mutations at codon 12 and codon 13. In 11 cases of gastric cancer, 4 showed point mutations: 1 at codon 12 and 3 at codon 13 of c-Ki-ras. In 3 pancreatic carcinomas, 2 had point mutations: one at codon 12 and the other at codon 13 of c-Ki-ras. The results indicate that c-Ki-ras gene point mutations are involved in the tumorigenesis of the human gastrointestinal system.

Adult↗

Full-length sequence of the cDNA for human erythroid beta-spectrin.

Spectrin is the major molecular consituent of the red cell membrane skeleton. We have isolated overlapping human erythroid beta-spectrin cDNA clones and determined 6773 base pairs of contiguous nucleotide sequence. This includes the entire coding sequence of beta-spectrin. The sequence translates into a 2137 amino acid, 246-kDa peptide. beta-Spectrin is found to consist of three distinct domains. Domain I, at the N terminus, is a 272-amino acid region lacking resemblance to the spectrin repetitive motif. Sequences in this region exhibit striking sequence homology, at both nucleotide and amino acid levels, to the N-terminal "actin-binding" domains of alpha-actinin and dystrophin. Between residues 51 and 270 there is 55% amino acid identity to human dystrophin, with only four single amino acid gaps in alignment. Domain II consists of 17 spectrin repeats. Several sequence variations are observed in typical repeat structure. Homology to alpha-actinin extends beyond domain I into the N-terminal portion of domain II. Domain III, 52 amino acid residues at the C terminus, does not adhere to the spectrin repeat motif. Combining knowledge of spectrin primary structure with previously reported functional studies, it is possible to make several inferences regarding structure/function relationships within the beta-spectrin molecule.

Amino Acid Sequence↗

Eosinophilic gastroenteritis with eosinophilic ascites: report of a case.

Eosinophilic gastroenteritis is a relatively uncommon disease of unknown etiology. Eosinophilic ascites resulting from significant serosal involvement is the rarest clinical subtype. The case reported here is of a 30-year-old male presenting with abdominal pain, diarrhea, and ascites. His personal history included childhood asthma, allergic rhinitis, and recurrent urticaria. The clinical picture was characterized by peripheral eosinophilia and eosinophilic infiltrates of the stomach and small bowel. Computed tomogram (CT) of the abdomen showed generalized thickening of the gastric and small bowel wall. Paracentesis revealed exudative ascites rich in eosinophils. The patient experienced an impressive response to steroid therapy.

Adult↗

Detection of beta-globin gene from single hairs.

DNA can be extracted from hair shafts and hair roots. The content of DNA in hairs is usually limited: the root end of hairs may contain 0.5 micrograms DNA and shed hairs contain less than 10 ng DNA. DNA analysis with restriction fragment length polymorphism (RFLP) requires microgram amounts of DNA. Such DNA cannot be obtained from such samples as single hairs or blood stains. Even with a little amount of DNA, specific genes of DNA can be greatly amplified to more than 10(6)-fold in vitro by polymerase chain reaction (PCR). We extracted the DNA from a half, 1, and 2 hair roots and 4 hair shafts and amplified the DNA with primer pairs (5'-GCACCATTCTAAAGAATAAC-3', 5'-GGATTGTAGCTGCTATTAGC-3') of the beta-globin gene, covering part sequences of the IVS-2 region by using the polymerase chain reaction for 50 cycles. The electrophoresis of the PCR product revealed a 131 base pairs band. Finally, we hybridized the PCR product with an IVS-2 probe (5'-GGGTTAAGGCAATAGCAAT-3'). A fragment of the IVS-2 extending from nucleotide 612 to 742 of the beta-globin gene can be demonstrated in the slot blot filter of hair roots and hair shafts, but not in the 1 microgram of blood which was not amplified by PCR. Detection of the beta-globin gene from single hairs by using the PCR may be useful for the diagnosis of thalassemia from single hairs.

Base Sequence↗

Waldenström's macroglobulinemia with unilateral massive pleural effusion: report of a case.

One rare case of Waldenström's macroglobulinemia with massive right-sided pleural effusion is reported. A 66-year-old woman was admitted to our hospital because of exertional dyspnea and chronic cough of three months' duration. Chest X-ray examination showed massive right-sided pleural effusion. Routine blood examination revealed severe anemia. Sternal bone marrow aspirate revealed a hypercellular marrow with replacement of 50% of the normal hematopoietic cells by a spectrum of lymphoid cells, including small lymphocytes, a few plasma-cytoid lymphocytes and immature plasma cells. The diagnosis was confirmed by serum protein electrophoresis and immunoglobulin assay. The effect of chemotherapy was satisfactory in the first two years as pleural effusion subsided completely and the marrow returned to near normal. But, the response was poor during the last 6 months. She died about 4 years after her first admission.

Aged↗

Infectivity in asymptomatic hepatitis B carriers: its correlation with HBeAg/anti-HBe status.

Sera collected from 190 asymptomatic HBsAg carriers with varying status of HBe markers were tested for HBV DNA using the slot-blot hybridization method and the results were compared with serum alanine aminotransferase (ALT) levels. No significant difference was observed in the positive rate of serum HBV DNA between patients showing either HBeAg (+)/normal ALT or HBeAg (+)/high ALT. On the other hand, in cases with positive anti-HBe or negative for both HBeAg and anti-HBe, statistically significant differences could be shown in HBV DNA positivity between normal and high ALT subjects. In the group of 100 patients possessing anti-HBe antibody, 50% of the cases with high ALT levels were positive for serum HBV DNA, whereas all of the cases with normal ALT levels were negative for serum HBV DNA (p less than 0.0002). In 21 patients showing HBeAg (-)/anti-HBe(-), HBV DNA could be detected in the serum of 60% of the cases with high ALT levels, but in none of the cases with normal ALT levels (p less than 0.05). Our results suggest that HBeAg alone is a reliable marker in the prediction of infectivity in asymptomatic carriers. In cases showing anti-HBe (+), loss of infectivity could be ascertained if the patient had normal serum ALT, but, continuous viral replication should be suspected in cases with high serum ALT values.

Adolescent↗

Molecular analysis of hemoglobin H disease in Taiwan.

The molecular basis of seven Chinese patients in Taiwan with hemoglobin H disease was investigated and was found to be heterogeneous in the mutation type. They were alpha-thalassemia-1 mutation combined with hemoglobin Constant Spring, an undetermined nondeletion form of alpha-thalassemia and a deletion form of alpha-thalassemia-2 mutations. The alpha-thalassemia-1 mutation was shown to be the --SEA type I haplotype.

Adult↗

[Campylobacter pyloridis in active chronic gastritis and peptic ulcer].

From Oct. 1986 to Jan. 1987, a total of 74 patients presenting for endoscopy were studied. These included 30 cases of active chronic gastritis and 44 cases of peptic ulcer. Biopsy specimens were taken during the endoscopy and sent to the laboratory for Campylobacter pyloridis culture within 2 hours. The culture was done by inoculated with chocolate agar, Brucella agar and 0.2% urea broth for urease activity. Results showed that, 53% (16/32) of active chronic gastritis, 77% (10/13) of gastric ulcer and 84% (26/31) of duodenal ulcer were positive for Campylobacter pyloridis. The specificity and sensitivity of urease positive rate are 64% and 90% respectively. It is higher as compared with bacteria culture. In addition, we found that 10 days would be needed for the routine culture and identification of this organism. But it took only 30 minutes to 6 hours for urease activity test. Therefore, we suggested that urease activity test could be a rapid diagnostic method for detecting Campylobacter pyloridis.

Campylobacter↗