Prenatal molecular diagnosis of four fetuses at high risk for X-linked adrenoleukodystrophy / 中华妇产科杂志

Objective To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy(X-ALD).Methods The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells.Maternal contamination was evaluated by paternity test.PCRRFLP,sequencing and denaturing high performance liquid chromatography(DHPLC)were used to detect the ABCD1 gene of fetal genome.Results In the pedigree 1,the PCR product(799 bp)of the fetus 1 and her father(normal control)could be digested with BcnI. No P560L mutation,which was present in the index patient,was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing.In the pedigree 2,the PCR product(232 bp)of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation,which was present in the propositus,was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing.In the pedigree 3,the PCR product(271 bp)was digested with AciI.the pattern of the fetus 3 and the propositus being the same,and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing.In the pedigree 4,the PCR product(269 bp)was analyzed with the DHPLC,and the pattern of elution peaks of the fetus 4 and her father was similar,but different from that of the propositus.No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing.Judging from the sex of the fetuses,fetuses 1 and 2 were normal homozygotes,fetus 3 was an ALD hemizygote,and fetus 4 was a normal hemizygote.Conclusion A new protocol for X-ALD prenatal molecular diagnosis is proposed,which would ensure the accuracy of prenatal diagnosis.

Characteristics of the cerebral vascular hemodynamic changes in acute stroke patients / 中国组织工程研究

BACKGROUND: The understanding about characteristics and variation trends of cerebral vascular hemodynamic indexes in high-risk stroke population,in prior and post-stroke period patients as well as in normal people,particularly the cerebral vascular hemodynamic index(CVHD) prior to stroke can improve diagnosis efficiency of stroke and increase sensitivity and specificity of stroke diagnosis.OBJECTIVE: To study the features of CVHD in acute stoke,in order to provide evidences for early evaluation and rehabilitation intervention.DESIGN: Case analysis and observational study based on patients.SETTING: A neurological department in a general hospital of military area command of Chinese PLA and a prevention and cure institute of cerebrovascular diseases.PARTICIPANTS: Totally 55 (male 38,female 17) inpatients in the Neurological Department of Fuzhou General Hospital of Chinese PLA from June to December 2001 were enrolled in the study.METHODS: CVHD was measured during the first week after onset and the data were analyzed.MAIN OUTCOME MEASURES: The CVHD,blood pressure and cerebral vascular hemodynamic parameters.RESULTS: The CVHD of each patient is below 75,median 32.5. The mean velocity,blood flow,maximum velocity,minimum velocity and difference between diastolic and critical pressures were lower than corresponding ones of healthy men in seventies( P ＜ 0.01 ). And the pulse wave velocity,peripheral resistance,specific impedance and critical pressure increased( P ＜ 0. 01).CONCLUSION: During the first week after stroke,the CVHD decreases and many hemodynamic parameters alters significantly. Examination of CVHD can reveal cerebral circulation insufficiency or predict an attack of cerebral stroke.

Study on a test of screening to predict stroke-using cerebral vascular hemodynamic indexes / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate efficacy and optimal cut-off-point through cerebral vascular hemodynamic indexes (CVHI) examination to predict stroke.</p><p><b>METHODS</b>A number of 20,333 people at 35 years old and over were checked by CVHI and accumulative score was calculated according to the value of detected indexes. Risk factors of stroke were investigated simultaneously. One hundred and sixty-eight patients with stroke occurred during 4-year following up. Typical syndromes and signs stroke were used as golden standard to evaluate screening efficacy of CVHI.</p><p><b>RESULTS</b>Score of optimal cut-off-point of cerebral vascular hemodynamic indexes was under 75 in ROC curve analyses. Sensitivity, specificity, accuracy, positive and negative predictive values, positive and negative likelihood ratios as well as Youden's index for predicting stroke within 4 years after examination were found to be 87.50%, 67.70%, 67.86%, 2.21%, 99.85%, 2.71, 0.18 and 0.55 respectively. Sensitivity and positive predict values for predicting cerebral vascular thrombosis were superior to predicting cerebral hemorrhage. Positive predicting value in risk exposure population was higher than that of overall population. Coefficiency of variation of cerebral vascular hemodynamic examination was 4.03%. The agreement rate of examination between two physicians was 97.62% and Kappa value was 0.94.</p><p><b>CONCLUSION</b>The score of optimal cut-off-point of cerebral vascular hemodynamic indexes examination was 75. Both Efficacy and reliability for predicting stroke seemed to be good, especially for predicting cerebral vascular thrombosis.</p>

Mutational analysis of seven Chinese pedigrees with X-linked adrenoleukodystrophy / 解放军医学杂志

Objective To identify the mutational genotype in seven Chinese families with X-linked adrenoleukodystrophy (X-ALD). Methods The coding region of ABCD1 gene of seven patients was amplified in 4 segments by PCR after reverse transcription using RT-PCR technology. The PCR products were purified and directly sequenced. To confirm the mutations, the genomic DNA was analyzed by PCR-restrictive digestion or direct sequencing of purified PCR products. Results Six base substitutions (709CA, 807GA, 1161CT, 2065CT, 2113TC and 2235CT), one base deletion (1801delAG) and one base insertion (1126 ins GCCATCG) were identified in seven X-linked adrenoleukodystrophy pedigrees, resulting in five missense mutations (A141T, R259W, P560L, L576P and R617C), two frame shift mutations (fs I246 and fs E471) and one nonsense mutation (S108X), respectively. Conclusion Four novel ABCD1 mutations, namely S108X, fs I246, R259W and L576P, were detected in Chinese X-linked adenoleukodystrophy patients. There was different ABCD1 gene mutation in different pedigree and no obvious correlation between the type of mutation and phenotype was found.