Objective　To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone- and joint- specific development-related genes.Methods　Total RNA of bone and joint were extracted with the modified single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. The double-stranded end-blunted cDNA were synthesized using TaKaRa's cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers.Results　A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed.Conclusion　The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue-specific genes.

OBJECTIVETo determine the genomic structure of low density lipoprotein receptor related protein 5 (LRP5) gene.

METHODScDNA sequence encoding LRP5 was used to screen genomic clones containing LRP5 gene by computer hybridization approach. By comparing the cDNA sequence of LRP5 with the genomic sequences, the genomic structure of LRP5 was determined, and then it was conformed by amplifying and sequencing the sequences of exons and splicing junction.

RESULTSThe genomic sequence of LRP5 gene was 131.6 kb in length, containing 23 exons and 22 introns. Three single nucleotide polymorphisms were detected within the coding sequences of LRP5 gene, namely A459G in exon 2, C2220T in exon 10 and G4416C in exon 21. Four polymorphic markers, D11S1917, D11S4087, D11S1337 and D11S4178, located in the 5' flank sequence, introns 1, 4, and 13 of the LRP5 gene, respectively.

CONCLUSIONThe characterization of genomic structure of LRP5 gene allows the investigators to detect disease-causing mutation within the gene and further study the function of LRP5 gene.

Base Sequence ; DNA ; chemistry ; genetics ; Exons ; Genes ; genetics ; Humans ; Introns ; LDL-Receptor Related Proteins ; Low Density Lipoprotein Receptor-Related Protein-5 ; Polymorphism, Single Nucleotide ; Receptors, LDL ; genetics ; Sequence Analysis, DNA

OBJECTIVETo determine the linkage between Smith-Fineman-Myers syndrome (SFMS) and X-linked nuclear protein(XNP) locus.

METHODSPolymerase chain reaction and denaturing polyacrylamide gel electrophoresis were used to genotype two polymorphic short tandem repeats within XNP gene.

RESULTSOne of the two short tandem repeats was informative in SFMS family from Shandong, China. Recombination between SFMS locus and XNP gene was observed in the SFMS family.

CONCLUSIONXNP gene is not associated with the disease in the SFMS family from Shandong, China. SFMS exhibits locus heterogeneity at molecular level.

Abnormalities, Multiple ; genetics ; Craniofacial Abnormalities ; genetics ; DNA Helicases ; Female ; Genetic Linkage ; Growth Disorders ; genetics ; Humans ; Intellectual Disability ; genetics ; Male ; Muscle Hypotonia ; genetics ; Nuclear Proteins ; genetics ; Pedigree ; Phenotype ; Polymorphism, Genetic ; Recombination, Genetic ; Syndrome ; X Chromosome ; X-linked Nuclear Protein

Objective　To analyze the genetic polymorphism of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 in Chinese Hans. Methods　Polymerase chain reaction-single strand length polymorphism(PCR-SSLP) was used to genotype 100 randomly selected individuals of the Han nationality at 8 STR loci(D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359) on chromosome 11 and 5 STR loci(D19S247, D19S714, D19S433, D19S246, D19S254) on chromosome 19. Results　Eight alleles and 26 genotypes, 9 alleles and 15 genotypes, 6 alleles and 16 genotypes, 12 alleles and 40 genotypes, 6 alleles and 19 genotypes, 12 alleles and 48 genotypes, 8 alleles and 20 genotypes, 7 alleles and 13 genotypes were observed at D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359. The heterozygosities for the 8 STR loci were 87%, 68%, 73%, 92%, 71%, 86%, 75% and 71%, respectively. Ten alleles and 19 genotypes, 10 alleles and 26 genotypes, 10 alleles and 24 genotypes, 11 alleles and 29 genotypes, 8 alleles and 18 genotypes were observed at D19S247, D19S714, D19S433, D19S246, D19S254. The heterozygosities for the 5 STR loci were 63%, 82% 72%, 81% 74%, respectively. Conclusion　 The distribution of allele frequencies of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 was consistent with the Hardy-Weinberg equilibrium and the highly genetic polymorphism was observed in Chinese Han population.

OBJECTIVETo map the gene responsible for nonsyndromic hearing impairment in a consanguineous family.

METHODSFirstly, X chromosome scanning was used to exclude X chromosome. Secondly, candidate gene analyzing and genome scanning were performed by homozygosity mapping. Then, additional markers flanking the tightly linked marker were tested to confirm linkage and decide the candidate region.

RESULTSThe nonsyndromic hearing impairment of this family was autosomal recessive. Twenty-five known genes were excluded. Autosomal genome scanning indicated that D17S1293 was tightly linked with disease gene. And further study mapped the disease gene to a 5.07 cM interval bounded by D17S1850 and D17S1818.

CONCLUSIONThe disease gene of the family is mapped to a 5.07 cM interval between D17S1850 and D17S1818, which is a new locus of autosomal recessive nonsyndromic hearing impairment.

Chromosome Mapping ; methods ; Chromosomes, Human, Pair 17 ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; Chromosomes, Human, X ; genetics ; Consanguinity ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Hearing Loss, Sensorineural ; genetics ; Humans ; Male ; Microsatellite Repeats ; Pedigree

OBJECTIVETo study the prevalence of methylenetetrahydrofolate reductase (MTHFR) C677T genotype and its association with deep vei n thrombophilia in Chinese.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was conducted to examine mutation with 63 deep vein thrombophilic patients and 80 health controls in Shandong Hans. The genotype frequencies were calculated by gene counting in patients and controls, and an analysis was made on the association of MTHFR C677T mutation with deep venous thrombosis in Shandong Hans.

RESULTSIn case- controls, the frequencies of C/T heterozygote were 41.27% and 43.75%; whereas those of T/T homozygote were 52.38% and 36.25%. Significantly elevated mutation was observed in patients(Chi-square=6.372, P 0.01 OR(T/T)=4.552 95% confidence interval:1.440-14.390, Chi-square =6.742 P=0.009).

CONCLUSIONThe C677T mutation of methylenetetrahydrofolate reductase gene is a risk factor associated with deep vein thrombophilia in Shandong Hans.

China ; DNA ; genetics ; Gene Frequency ; Genotype ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; Odds Ratio ; Oxidoreductases Acting on CH-NH Group Donors ; genetics ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Thrombophilia ; enzymology ; genetics ; Venous Thrombosis ; enzymology ; genetics

OBJECTIVETo analyze clinical features and mutation in MYH9 gene for a family featuring autosomal dominant May-Hegglin anomaly.

METHODSClinical and pathological features of all family members were analyzed. Blood samples were collected from the proband and other family members, and genomic DNA was extracted. Potential mutations of MYH9 gene exons 10, 25, 26, 30, 38 and 40 were screened with PCR and direct sequencing. After a mutation was identified in the proband, other affected members as well as healthy members from this family were analyzed with a pair of primers to amplify the mutant site. The PCR products were digested with Taq I enzyme and analyzed with agarose gel electrophoresis.

RESULTSAll affected members had bleeding tendency and typical features including giant platelets, thrombocytopenia and characteristic Dohle body-like leukocyte inclusions. A heterozygous missense mutation c.5521G>A (p.Glu1841Lys) in exon 38 of the MYH9 gene was identified in all affected members from this family.

CONCLUSIONThe variant, c.5521G>A (p.Glu1841Lys) of MYH9, has co-segregated with the phenotype in the family. The mutant site is a hot spot in Chinese population.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Exons ; Female ; Genes, Dominant ; Hearing Loss, Sensorineural ; Humans ; Male ; Molecular Motor Proteins ; genetics ; Mutation ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Thrombocytopenia ; diagnosis ; genetics

Lung cancer, one of the most prevalent cancer types globally, underscores the critical importance of early detection and precise diagnosis in its management. However, conventional pathological diagnostic methods encounter various limitations, including the intricacies of the diagnostic process and the challenges in achieving uniform results. In contrast to traditional pathology, digital pathology integrates digitized pathological information with artificial intelligence, and offers a pathway for rapid and accurate diagnostic assistance. It holds the potential to delve into the tumor characteristics and microenvironment information, thereby supporting precision diagnosis and treatment for lung cancer. This article elucidates the recent applications of digital pathology in lung cancer diagnosis, staging, treatment, and prognosis. Additionally, it addresses the challenges currently faced by digital pathology.