OBJECTIVETo study the clinicopathological and molecular genetic characteristics of hereditary nonpolyposis colorectal cancer (HNPCC), to enable the early diagnosis and to evaluate the treatment.

METHODSWe analyzed 12 families of HNPCC from Wenzhou, Zhejiang province, China. Mismatch repair genes hMSH2 and hMLH1 expression and microsatellite instability of tumor tissue were studied using microdissection, microsatellite analysis, immunohistochemical staining and Gene Scan analysis. Direct DNA sequencing of hMSH2 and hMLH1 were performed subsequently.

RESULTSAltogether 32 patients with colorectal cancer were recognized in 12 HNPCC families, with the median age of 45.2 years (75.0% before the age of 50 years). The proximal tumors accounted for 51.1%, while multiple colorectal cancers accounted for 34.4%. Poor differentiation cancers occupied half of the patients (53.1%). And 68.8% of the patients had the tumor of Dukes A and B. Among 12 HNPCC families, 7 cases in 6 HNPCC families developed extracolonic cancer. 13 cases died during follow up of 1 - 23 years. The median survival time was 6.4 years. 19 alive cases followed up from 1 to 28 years. All tumors (9/9) displayed microsatellite instability, with the half losing hMSH2 or hMLH1 expression. In the 5 genetic analyzed kindreds 3 possessed germline mutation. Two of three mutations have not been reported in the worldwide database previously.

CONCLUSIONHNPCC showed distinct clinicopathological characteristics. Microsatellite instability analysis and immunohistochemical staining might be the effective screening methods before direct DNA sequencing for the detection of mutation in mismatch repair genes. It is important to analyze the members of affected families.

Adaptor Proteins, Signal Transducing ; Adult ; Aged ; Carrier Proteins ; China ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; metabolism ; pathology ; DNA Mutational Analysis ; DNA, Neoplasm ; chemistry ; genetics ; DNA-Binding Proteins ; analysis ; genetics ; Family Health ; Female ; Humans ; Immunohistochemistry ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; Mutation ; Neoplasm Proteins ; analysis ; genetics ; Nuclear Proteins ; Proto-Oncogene Proteins ; analysis ; genetics

OBJECTIVETo identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level.

METHODSIn two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family.

RESULTSA nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family.

CONCLUSIONThe mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.

Alternative Splicing ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; China ; Chromosomes, Human, X ; genetics ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Linkage ; Humans ; Male ; Membrane Transport Proteins ; Mutation ; Osteochondrodysplasias ; genetics ; pathology ; Pedigree ; Transcription Factors

AIMIdentification of the rodent counterparts of human and canine epididymal cDNAs HE3, HE4 and Ce8/Ly6G5C by sequence homology and analysis of their expression patterns and regulation level in the rat.

METHODS"Electronic screening" of Expressed Sequence Tag (EST) and genomic databases, followed by RT-PCR and Northern blot analysis.

RESULTSRodent ESTs and genomic sequences homologous to HE3, HE4 and Ce8/Ly6G5C were identified in the public databases and the "full-length" rat cDNAs cloned. To emphasise their homology to the human and canine genes, they were named Me3/Re3, Me4/Re4 and Re8 for mouse and rat counterparts, respectively. mRNA expression patterns were analysed in rats, including rat HE1 and HE5/CD52 counterparts as controls. Re3 and Re8 mRNAs were only found in the rat epididymis, while Re4 showed a broader tissue distribution. Within the epididymis, Re3 and Re4 mRNAs were detected in all regions; Re8, on the other hand, was restricted to the caput. During postnatal development, Re3 and control mRNAs were found from the earliest stages investigated, while Re8 mRNA was observed only from day 24 postnatum, corresponding to the onset of spermatogenesis in the prepubertal testis. Castration and testosterone supplementation of adult male rats suggested that none of the cloned mRNAs was directly androgen-regulated. Efferent duct ligation, however, showed that Re8 mRNA levels depended on testicular factors other than androgens.

CONCLUSIONThe novel rodent cDNAs can now be used to monitor epididymal gene expression more closely and to set up various regulatory and functional studies.

Amino Acid Sequence ; Animals ; Blotting, Northern ; Carrier Proteins ; chemistry ; genetics ; DNA, Complementary ; chemistry ; Dogs ; Epididymal Secretory Proteins ; chemistry ; genetics ; Epididymis ; chemistry ; Glycoproteins ; chemistry ; genetics ; Humans ; Male ; Molecular Sequence Data ; Proteins ; chemistry ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology ; Tissue Distribution ; beta-Defensins

To detect chromosome translocation t(11;18) (q21;q21) and the nuclear expression of bcl-10 in gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma in Chinese, a possible API2-MALT fusion transcript specific to t(11; 18) (q21; q21) in tumors from 42 cases of primary gastrointestinal lymphoma (29 cases of low grade MALT lymphoma, 13 cases of transformed MALT lymphoma) and 40 cases of diffuse large B cell lymphoma was examined by means of RT-PCR and proved by DNA-sequencing. Bcl-10 expression was examined by immunohistochemical method. The results showed that t(11;18) (q21;q21) was 14% positive in cases of low grade MALT lymphomas and 46% positive in transformed MALT lymphomas, but none in cases of DLBCL. Bcl-10 nuclear expression was seen 61% in low grade MALT and 69% in transformed MALT lymphoma. It was suggested that t(11;18) (q21;q21) was related to the prognosis and development of highly advanced MALT lymphoma but not relevant to DLBCL. Bcl-10 nuclear expressions were not significantly different between these two groups, which remains to be explained.
Adaptor Proteins, Signal Transducing ; B-Cell CLL-Lymphoma 10 Protein ; Carrier Proteins ; analysis ; Cell Nucleus ; chemistry ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 18 ; Humans ; Immunohistochemistry ; Lymphoma, B-Cell, Marginal Zone ; chemistry ; genetics ; Translocation, Genetic

OBJECTIVETo detect the trans-factors specifically binding to the strong enhancer element (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene.

METHODSYeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library to identify potential trans-factors that can interact with core sequence of GPEI(cGPEI). Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of transfactors to cGPEI.

RESULTScDNA fragments coding for the C-terminal part of the transcription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. The binding of c-Jun and ANT to GPEI core sequence were confirmed.

CONCLUSIONSRat c-jun transcriptional factor and ANT may interact with cGPEI. They could play an important role in the induced expression of GST-P gene.

Animals ; Biological Assay ; methods ; Carrier Proteins ; chemistry ; isolation & purification ; DNA Primers ; DNA, Complementary ; genetics ; Enhancer Elements, Genetic ; genetics ; Enzyme Induction ; Gene Expression Regulation ; Gene Library ; Glutathione Transferase ; genetics ; Lung ; Rats ; Sequence Analysis, DNA ; Yeasts

The present study was designed to investigate polymorphism in Duffy binding protein (DBP) gene of Plasmodium vivax isolates of Korea. Thirty samples were obtained from P. vivax patients in Yonchon-gun, Kyonggi-do in 1998. The PCR products of the samples were subjected to sequencing and hybridization analyses of the regions II and IV of P. vivax DBP gene. Two genotypes, SK-1 and SK-2, were identified on the basis of amino acid substitution and deletion. The genotype of 10 isolates was SK-1 and that of 20 isolates was SK-2. Most of the predicted amino acids in the region II of DBP gene were conserved between the Korean isolates and Belem strain except for 4-5 amino acid substitutions. In the region IV of DBP, a 6-bp insert that was shown in the Sal-1 allele type was found in SK-1, and a 27-bp insert that was shown in the Papua New Guinea allele type was found in SK-2. In conclusion, the present findings suggest that two genotypes of P. vivax coexist in the endemic area of Korea.
Amino Acid Sequence ; Animals ; *Antigens, Protozoan ; Base Sequence ; Carrier Proteins/*analysis/chemistry/*genetics ; DNA, Protozoan/genetics ; Genotype ; Human ; Korea ; Malaria, Vivax/parasitology ; Molecular Sequence Data ; Plasmodium vivax/*genetics/isolation & purification ; Polymerase Chain Reaction ; *Polymorphism (Genetics) ; *Protozoan Proteins ; Receptors, Cell Surface/*analysis/chemistry/*genetics ; Support, Non-U.S. Gov't

OBJECTIVECloning and characterization of a novel gene by exon trapping and exon linking at chromosome 8p22.

METHODSA novel gene was cloned using exon trapping and exon linking, and its expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot.

RESULTSA sequence containing 3 exons was found. The sequence is homologous with the putative gene AK024799 which consists of 2880 bp cDNA with 1362 bp open reading frame and codes 454 amino acids with an SH(2) domain. The gene was named SH(2)A at chromosome 8p22. SH(2)A gene is ubiquitously expressed in various tissues with three transcripts. The aberrant expression of SH(2)A gene in some cancers was detected.

CONCLUSIONSH(2)A is a novel docking protein of SH(2) signaling protein family, which may play an important role in cellular signal transduction. It relates to the pathogenesis of tumor.

Animals ; Blotting, Northern ; COS Cells ; Carrier Proteins ; Chromosome Mapping ; Chromosomes, Human, Pair 8 ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Exons ; Gene Expression ; Genes ; genetics ; Guanine Nucleotide Exchange Factors ; Humans ; Introns ; Membrane Proteins ; genetics ; Molecular Sequence Data ; RNA ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; rac1 GTP-Binding Protein ; src Homology Domains ; genetics

OBJECTIVETo identify the mutation of the methylmalonic aciduria (cobalamin deficiency) CblC type, with homocystinuria (MMACHC) gene in a pedigree with methylmalonic aciduria.

METHODSThe MMACHC gene mutation was detected using polymerase chain reaction (PCR) and DNA sequencing. The MMACHC gene of 50 healthy people was also sequenced as control.

RESULTSA new mutation of 146_154 del CCTTCCTGG was found in the patient and his father, and was absent in the controls.

CONCLUSIONA new mutation (146_154 del CCTTCCTGG) in the MMACHC gene was detected in a Chinese family with methylmalonic aciduria.

Amino Acid Metabolism, Inborn Errors ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Carrier Proteins ; chemistry ; genetics ; Case-Control Studies ; Child, Preschool ; DNA Mutational Analysis ; Exons ; genetics ; Fathers ; Female ; Humans ; Male ; Methylmalonic Acid ; metabolism ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Pregnancy ; Protein Structure, Secondary