Objective To evaluate the clinical utility of CMV pp65 antigenemia by CMV brite Kit for predicting active/reactive CMV infection S as well as of CMV diseases in bone marrow or peripheral stem cell transplant patients. We also investigated the efficacy of preemptive therapy guided by detection of CMV antigenemia. Methods A total of 210 EDTA anticoagulant plasma samples from 36 bone marrow or Peripheral Stem Cell Transplant Patients were prospectively collected from September 1999 to April 2000. The specific CMV antibody IgG/IgM of all patients were detected by ELISA. We detected CMV pp65 antigenemia by indirect immunofluorescence assay using CMV Brite Kit. All blood samples were detected weekly from week 3 after bone marrow transplantation until 100 days or antigenemia turning negative/dischage or death. Ganciclovir preemptive therapy was initiated at first positive pp65 antigenemia. Results Of 36 bone marrow or Peripheral Stem Cell Transplant Patients, 16 patients occurred positive pp65 antigenemia, 15 patients suffered from symptomatic CMV infections or CMV diseases. In 14 patients of positive pp65 antigenemia receiving gaciclovir therapy at first antigenemia, 2 patients died (mortality rate 14.2%), 12 patients of pp65 antigenemia became negative. Otherwise, 2 untreated cases died. The study showed a significant difference in mortality rate between treated and untreated patients (P

Objective To discuss curative results of breast-conserving therapy for early primary breast cancer. Methods Fifty-two patients with stage Ⅰ~Ⅱ breast cancer(solitary tumor ≤ 3 cm in size) underwent local lumpectomy and axillary lymph node dissection from March 2000 to September 2005 in this hospital.After operation,the radiation therapy,chemotherapy,and endocrine therapy were given.Results The breast specimens were pathologically examined and no infiltrating margin was found.The rate of good cosmetic effects was 86.5%(45/52).Follow-up examinations in 50 cases for 10~36 months(median,16 months) found no local recurrence and distant metastasis.Conclusions The cosmetic and clinical results of breast-conserving surgery for stage Ⅰ~Ⅱ primary breast cancer are satisfactory.

AIM:To investigate the regulatory effects of phosphatylinositol 3-kinase/protein kinase B (PI3K/PKB) signaling pathway on the expression of osteopontin ( OPN) in transforming growth factor-β1 ( TGF-β1 )-induced hu-man hepatic stellate cells .METHODS:Human hepatic stellate cell line LX-2 was cultured in DMEM and stimulated by TGF-β1 at the final concentration of 2.5, 5, 10 and 20 μg/L for 24 h or at final concentration of 10 μg/L for 12 h, 24 h and 48 h.LX-2 cells were pretreated with wortmannin , a specific inhibitor of PI3K/PKB signaling pathway , at final con-centration of 0.1 μmol/L for 1 h, followed by incubation with TGF-β1 at final concentration of 10μg/L for 24 h.The cells were collected.The expression of OPN was detected by real-time PCR and Western blotting .RESULTS: In LX-2 cells, the expression of OPN was apparently elevated when incubated with TGF-β1 .With the increase in TGF-β1 concentration or the extension of incubation hours , the expression of OPN was increased gradually in a dose-and time-dependent manner with certain limits.LX-2 cells pretreated with wortmannin and incubated with TGF-β1 had a significant decrease in the OPN expression as compared with control group (P<0.01).CONCLUSION:The expression of OPN in TGF-β1-induced LX-2 cells is regulated by the PI3K/PKB signaling pathway.

Objective To study the validity of Dynamic Occupational Therapy Cognitive Assessment for Children (DOTCA-Ch) on the cognitive function of children with cerebral palsy. Methods 30 children with cerebral palsy were assessed with DOTCA-Ch and Chinese Binet Test of Intelligence by trained rehabilitation medicine graduates within 1 week. The correlation between these tests were analyzed. Results The total DOTCA-Ch scores, spatial orientation and thought operation function were strongly correlated with IQ, while spatial perception,praxis and optomotor tissue were slightly correlated with IQ. Conclusion DOTCA-Ch can comprehensively assess the cognitive function of children with cerebral palsy.

OBJECTIVETo investigate the clinical and molecular genetics characteristics of a patient with mild androgen insensitivity syndrome (MAIS).

METHODSClinical data of the patient was collected, and DNA was isolated from peripheral blood sample. Eight exons of AR gene were amplified by PCR with specific primers and directly sequenced by Sanger method. The results were compared with standard sequences from GenBank. Online Polyphen-2 software was applied to predict the effect of mutation on the protein function and compare the conservation of the sequence at the mutation site in various species. The exon of the AR gene containing the mutated site was analyzed in 90 unrelated normal males using PCR and restrictive digestion with Sfa NI.

RESULTSSequence analysis has detected a novel missense mutation in codon 176 of exon 1 (Ser176Arg) of the AR gene. Analysis with polyphen-2 software has indicated the codon to be highly conserved across various species, and that the S176A mutation has caused damage to the protein structure and function (prediction score=0.999). The same mutation was not detected in 90 healthy males.

CONCLUSIONThe S176A mutation of the AR gene may contribute to the mild androgen insensitivity syndrome.

Adult ; Amino Acid Sequence ; Androgen-Insensitivity Syndrome ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Receptors, Androgen ; genetics

OBJECTIVETo assess the association of PEX10 gene and 1p36 copy number variations in 1p36 region with concurrent epilepsy through analyzing 3 cases.

METHODSThe karyotypes of 3 patients were determined by high resolution chromosome banding, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) combined with single nucleotide polymorphism array (SNP) technology. Real-time PCR was carried out to determine the mRNA levels of PEX10 gene in peripheral blood of the patients.

RESULTSNo abnormality was found upon high resolution karyotyping. MLPA analysis showed that all of the 3 patients had a copy number variation of subtelomeric region in the short arm of chromosome 1, which was confirmed by FISH and SNP chip analyses. Case 1 and case 2 both had an epilepsy phenotype, and their copy number variations have encompassed the PEX10 gene. On the other hand, case 3 has absent epilepsy, and its PEX10 gene copy number was normal. Family investigation confirmed that the chromosome abnormalities in all of the 3 cases were of de novo type. Compared with healthy controls, real-time PCR showed that mRNA of the PEX10 gene was increased in case 1 but decreased in case 2.

CONCLUSIONThe abnormal expression of PEX10 gene resulting from copy number variations of 1p36 region may be associated with the epilepsy phenotype.

Child ; Chromosomes, Human, Pair 1 ; DNA Copy Number Variations ; Epilepsy ; genetics ; Female ; Humans ; Peroxins ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptors, Cytoplasmic and Nuclear ; genetics

OBJECTIVETo investigate the phenotype-genotype association of isodicentromere Y chromosome by analysis of two female patients carrying the chromosome with sexual development disorders.

METHODSThe karyotypes of the two patients were determined by application of conventional G banding of peripheral blood samples and fluorescence in situ hybridization (FISH). PCR was applied to detect the presence of SRY gene.

RESULTSConventional karyotype analysis showed case 1 to be a mosaic: mos.45,X[38]/46,X,+mar[151]/47,XY,+mar[5]/47,X,+mar × 2[2]/46,XY[4], FISH showed that 12 different cell lines were presented in the karyotype of case 1 and partial cell lines with SRY gene, the marker is an isodicentromere Y chromosome [idic(Y)(p)]. No mutation was found in the SRY gene. The karyotype of case 2 was mos.45,X[25]/46,X,+mar[35]. FISH showed the marker to be an idic(Y)(p) without the SRY gene.

CONCLUSIONThe karyotype of patients carrying idic(Y)(p) seems unstable, and female patients have the characteristics of short stature and secondary sexual hypoplasia. Karyotype analysis combined with FISH analysis can accurately determine the breakpoint of idic(Y) and identify the types of complex mosaic, which may facilitate genetic counseling and prognosis.

Adolescent ; Child ; Chromosomes, Human, Y ; Disorders of Sex Development ; genetics ; Female ; Humans ; Karyotype ; Sex Chromosome Aberrations ; Sex-Determining Region Y Protein ; genetics

OBJECTIVETo determine the molecular etiology for a muscular dystrophy pedigree with target region sequencing platform using hereditary myopathy capture array.

METHODSSpecific gene testing was performed based on the clinical diagnosis. Since no pathogenic mutation was found, target region sequencing with hereditary myopathy capture array combined with Sanger sequencing and bioinformatics analysis were employed in turn. PolyPhen and NCBI were used to evaluate the pathogenicity of identified mutation and conservation of the gene.

RESULTSTarget region sequencing indicated the proband has carried a heterozygous c.3353 A>C mutation of COL6A3 gene, which was confirmed by Sanger-sequencing in 4 affected individuals from the family. The same mutation was not detected in healthy members of the pedigree. Bioinformatics analysis suggested that the mutation has caused a highly pathogenic amino acid substitution from Histidine to Proline. The affected patients featured normal intelligence with mild myogenic damage by muscle biopsy, slightly increased serum creatine kinase and slow disease progression, which was consistent with Bethlem myopathy.

CONCLUSIONTarget region sequencing is an effective and efficient method for genetic testing. The heterozygous c.3353A>C mutation in exon 8 of the COL6A3 gene probably underlies the Bethlem myopathy with autosomal dominant inheritance.

Adult ; Amino Acid Sequence ; Amino Acid Substitution ; Base Sequence ; Collagen Type VI ; genetics ; Contracture ; genetics ; Exons ; Female ; Heterozygote ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Muscular Dystrophies ; congenital ; genetics ; Mutation, Missense ; Pedigree ; Young Adult

Objective: To investigate the effects of fiber surface deposited with silicon dioxide films by atomic layer deposition on properties of dental fiber-reinforced composites. Methods : SiO2 films were deposited on the surface of quartz fiber by atomic layer deposition(ALD). Then the quartz fiber was used to manufacture fiber resin composites, which were divided into four groups: A(no soaking agent removal), B(soaking agent removed), C(soaking agent removed and silanization), and D(soaking agent removed, 600 ALD cycles performed and then silanization). Scanning electron microscopy, water contact angle test, hygroscopicity test, CCK8 test and three-point bending test were used to investigate the properties of fiber resin composites. Results: The surface morphology of the quartz fiber treated by ALD was smooth and had no obvious change compared with that before treatment. Moreover, the quartz fiber showed hydrophobicity after silanization. The results of three-point bending test revealed that the mechanical properties of fiber-resin composites modified by ALD were significantly improved(P<0.05). When viewed by scanning electron microscopy, a good interfacial bonding could be seen between quartz fibers and the resin matrix in Group D. In addition, it was found that Group D had low absorbability, low solubility and good biocompatibility. Conclusion It is shown that deposition of SiO2 films on the quartz fiber by ALD can significantly enhance the mechanical properties of fiber-reinforced composites.