Pathogenic gene mutation in a Han Chinese family with hereditary vitreous amyloidosis identified by Sanger sequencing / 临床检验杂志

Objective Our purpose was to investigate the pathogenic gene mutation of a Han Chinese family with vitreous amyloidosis.Methods The 9 individuals(proband,1 affected member and 7 unaffected members) of the family were selected and their DNA was extracted from peripheral blood.The 4 exons of transthyretin(TTR) gene were amplified by polymerase chain reaction(PCR) technique.The amplified products of TTR gene were sequencing by Sanger technique.We also selected 100 unrelated healthy individual as the control group.Results By DNA sequencing,a heterozygous mutation was found in 4 of the 9 subjects from the family.The transition of adenine to cytosine(AAG ＞ ACG) was detectable in exon 2 of TTR,which changed the amino acid composition at codon 35 (Lys35Thr).This mutation did not presented in control group.Conclusion The heterozygosis mutation of TTR gene Lys35Thr should be a pathogenic mutation for the family with vitreous amyloidosis.

A novel splicing mutation in intron 2 of DSPP gene in a family with dentinogenesis imperfecta type Ⅱ / 临床检验杂志

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

Construction of recombinant expression vector and prokaryotic expression of human epididymal sperm protein P34H / 中华男科学杂志

<p><b>OBJECTIVE</b>To acquire purified recombinant human epididymal sperm protein P34H for basic and clinical studies.</p><p><b>METHODS</b>On the basis of cloning of P34H coding region, P34H fragment was subcloned into the pQE-30 expression vector. The recombinant expression vector designated pQE-30/P34H was transformed into E. coli to induce the expression of the recombinant protein P34H on the reduction of IPTG. After sonication, the recombinant protein P34H was purified from the supernatant with Ni-NTA resin under native conditions. It was identified by SDS-PAGE analysis and DNA sequencing.</p><p><b>RESULTS</b>Recombinant expression vector pQE-30/P34H was correctly constructed, identified with PCR and double-enzyme digestion. And the results of SDS-PAGE analysis and DNA sequencing showed that the protein was what we had hoped to acquire.</p><p><b>CONCLUSION</b>Purified recombinant P34H can be acquired successfully with the above mentioned prokaryotic expression method.</p>

COL2A1 gene mutation of a family with spondyloepiphyseal dysplasia conginita / 医学研究生学报

Objective:To investigate a large Chinese family in which 9 patients over 4 generations were diagnosed with a form of autosomal dominant spondyloepimphyseal dysplasia(SEMD).Mothods:X-Ray radiograph of proand at 18-month showed absence of secondary ossification centra of femoral heads.His father at 24-year presented severe spondyloepiphyseal changes that principally involved the vertebral bodies,the femoral necks and femoral heads and characterized by generalized platyspondyly with thoracolumbar scoliosis,irregular femoral necks,absent ossification of femoral heads,flat acetabular roofs and coxa vara.The other patients had similar clinical and radiological features.Haplotyping was performed with leukocyte DNA for 5 micosatellite repeat markers from chromosome 12 and the result showed COL2A1 gene as a candidate gene.A total of 54 exons and promoter of COL2A1 gene were amplified and sequenced from all patients and available normal relatives.In addition,exon 23 of COL2A1 gene was amplified and sequenced from 10 controls simultaneously.Results:All patients were identified a 1510(G→A) transition in exon 23 of COL2A1 gene that caused a change from a COL2A1 coding region in available glycine to serine at amino acid position 504.No mutation was found in the normal relatives and 10 controls. Conclusion:The mutation of COL2A1 gene is responsible for this form of SEDC of the family.This is the first familial report of SEDC relating to 1510G→A mutation of COL2A1 gene.The detailed clinical radiogram data will be useful for extending the phenotypic spectrum of type Ⅱcollagenopathies.

Progress in studies on molecular genetics of spondyloepiphyseal dysplasia / 医学研究生学报

Spondyloepiphyseal dysplasia(SED) includes a group of disorders that cause deformation of vertebrae and epiphyses following gene mutations.Its main clinical manifestations are short stature(with a disproportionately short-trunk),chest malformation and early-onset joint degeneration.These disorders are broadly categorized into two subtypes: congenita(SEDC) and tarda(SEDT).In the 2006 revision of the International Nosology and Classification of Genetic Skeletal Disorders,372 different conditions were listed,of which 215 were associated with one or more of 140 different genes.SEDC has consistently been shown to correlate with defects in the gene COL2A1 on the long arm of chromosome 12,whose product is needed to form normal type-Ⅱ collagen.The gene responsible for SEDT is SEDL,mapped to the short arm of the X chromosome(Xp22).This paper briefly reviews the progress in the studies of molecular genetics of SEDC,SEDT and other rare forms of SED,which might provide some practical help for genetic and prenatal diagnoses of SED.

A novel RNA-splicing mutation in COL1A1 gene causing osteogenesis imperfecta typeⅠin a Chinese family / 医学研究生学报

A) in COL1A1 gene resulting in OI in a Chinese family. The detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in OI and exploring the phenotype-genotype correlations in OI.

An infertile 45,X male carrying an unbalanced(Y,13) translocation:A clinical cytogenetic and molecular study / 医学研究生学报

Objective: To report a case of azoospermia with a karyotype of 45,X,der(Y)t(Y;13)(q11.2;q12),-13,accompanied with slight bilateral gynecomastia and multiple nodules.Methods: The karyotype was identified by karyotyping and FISH,and the breakpoints of the Y chromosome and the copy number of the BRCA2 gene in 13q12 determined by PCR-STS and DNA polymorphic analysis.The testis and nodule tissues of the patient were obtained for biopsy.Results: FISH confirmed SRY and centromere of the Y chromosome on the questionable 13 chromosome and the karyotype to be 45,X,der(Y)t(Y;13)(q11.1;q12),-13.ish der(Y)(SRY+,DYZ3+,wcp13+).PCR-STS showed the deletion of regions AZFa,b and C,with a breakpoint located inYq11.1 below sY82.No deletion of the BRCA2 gene was observed.The patient was diagnosed with Sertoli cell-only syndrome by testicular biopsy and with angiolipomata by pathological examination of the nodule tissue.Conclusion: The patient's phenotype of complete masculinization could be attributed to presence of the SRY gene,and his azoospermia with small testis to the absence of a fragment from Yq11.1 to Yqter.However,the molecular mechanism of angiolipoma remains unknown.

Intratracheal medication by aerosol rebreathing method in emergency treatment of acute respiratory tract injury caused by inhalation of phosphorus trichloride / 医学研究生学报

Objective: To explore the emergency treatment of acute respiratory tract injury caused by inhalation of phosphorus trichloride.Methods: The clinical data of 16 patients with acute respiratory tract injury caused by inhalation of phosphorus trichloride were analyzed.Intratracheal inhalation of salbutamol sulfate solution and pulmicort repules atomized liquid by aerosol rebreathing method was performed immediately after the diagnosis was made.Anti-inflammatory,fluid replacement,and other symptomatic treatment were given at the same time.Treatment lasted for 3-5 days.Results: The symptoms and signs improved significantly after 3-5 times of inhalation.Among the 16 patients,15 were cured and one improved;the cure rate was 93.75%.The shortest observation period was 2 days and the longest 7 days with an average period of 2.38 days.All cases were followed up for 3 months.No complications were found except one with pleural thickening.Conclusion: Early treatment of acute respiratory tract injury with intratracheal corticosteroid and?2 receptor activator is satisfactory for patients with inhalation of phosphorus trichloride.The method is fast-acting,efficient,with less side-effects,convenient,and easily accepted by patients.

Influence of all-trans retinoic acid on the invasive potential of epithelial ovarian carcinoma cell line COC2 / 医学研究生学报

Objective: To observe the influence of all-trans retinoic acid(ATRA) on the expressions of E-cadherin,heparanase and VEGF in epithelial ovarian carcinoma cell line COC2,and to investigate the anti-metastatic potential and possible action mechanism of ATRA.Methods: We used flow cytometry to examine the expressions of E-cadherin,heparanase and VEGF proteins in the epithelial ovarian carcinoma cell line COC2 treated with different concentrations of ATRA.Results: ATRA significantly increased the expression of E-cadherin and decrease that of VEGF and hepareanse in a dose-dependent manner.Conclusion: ATRA can inhibit cell proliferation,improve cell-cell adhesion and downregulate the expressions of VEGF and heparanse proteins,suggestive of an anti-angiogenic and anti-metastatic potential.

Relationship between DNA ploidy heterogeneity and clinical biological behavior on patients with malignant tumors / 医学研究生学报

Objective:To explore the relationship between DNA ploidy heterogeneity and clinical biological behavior on patients with malignant tumor.Methods:The DNA ploidy heterogeneity of tumor tissue was measured in 163 patients with malignant tumors by flow cytometry.The relations were analyzed between DNA ploidy heterogeneity and clinical stage,pathological grade,metastasis rate of patients with malignant tumors.Results:The rates of DNA ploidy heterogeneity were significantly different in different tumors.The rates of heterogeneity raised with increase of clinical stage and pathological grade(P

Prenatal molecular diagnosis of two pregnancies in familial G504S mutation of COL2A1 gene resulting spondylepiphyseal dysplasia congenita / 临床检验杂志

Objective To report the prenatal molecular diagnosis for two gravida in a family with spondylepiphyseal dysplasia congenita(SEDC)caused by G504S mutation of COL2A1 gene.Methods DNA of the two fetuses was extracted from amniotic fluid at the 19+3 and 18+6 weeks of gestation respectively.Direct sequencing of two samples were performed after amplifying exon 23 of COL2A1 containing the potential mutation.The femur length and biparietal diameter of the first fetus were measured by sonographic scans every two weeks from 17+3 weeks to 27+3 weeks of gestation,and for the second fetus these parameters were measured from 16+1 to 19+1 weeks of gestation.Results Sequncing analysis revealed the first fetus and his mother presented the same mutation which is specifically associated with SEDC,but the second fetus did not show the mutation of COL2A1 gene.Biparietal diameters of the both fetuses were appropriate for gestational age.Femur length of the second fetus was normal for gestational age but that of the first fetus was shortened evidently after the 23 week of gestation.The parents of the first fetus determined to terminate the pregnancy.A medical termination was carried out at 27+5 weeks of gestation and a male fetus with a relatively large head and short limbs was delivered.The radiological findings of the fetus were consistent with SEDC including generalized platy spondesand shortened long bones.Conclusions Prenatal molecular diagnosis is important for the fetus with risk of SEDC and useful for genetic counseling.Genotype of fetus with risk of SEDC can be identified before sonographic scan.Molecular genetic analysis in conjunction with sonographic monitoring was helpful in prenatal diagnosis of SEDC.