The Gene Polymorphism of VMAT2 Is Associated with Risk of Schizophrenia in Male Han Chinese

Objective@#To investigate the association between gene polymorphism of vesicular monoamine transporter type 2(VMAT2) and schizophrenia in Han Chinese population. @*Methods@#430 patients with schizophrenia and 470 age-sex matched controls were recruited from four mental health centers. All patients were diagnosed by two psychiatrists based on the Structured Clinical Interview for DSM Disorders (SCID). The ligase detection reactions (LDR) method was used to assess the polymorphism of the two SNPs (rs363371 and rs363324) of VMAT2. @*Results@#No associations of two SNPs with schizophrenia was found. When we stratified males and females for the analysis, we found that that in the recessive model of rs363371, there was an obvious significant association between rs363371 and schizophrenia in males (OR=0.564, 95% CI=0.357–0.892, p=0.014) but not females. For the association between rs363324 and schizophrenia, no association was found in either males or females. No association was found when stratifying early-onset schizophrenia and late-onset schizophrenia. @*Conclusion@#Our findings indicate that both rs363371 and rs363324 were not associated with schizophrenia, while it seemed that the AA genotype of rs363371 plays a protective effect in male Chinese in developing schizophrenia.

Association between adverse obstetric outcomes and abnormal maternal serum markers in the second trimester screening / 上海交通大学学报（医学版）

Objective · To evaluate the association between the abnormal maternal serum markers of alpha fetoprotein (AFP), human chorionic gonadotrophin (hCG) and unconjugated estriol (uE3) in the second trimester screening and the adverse obstetric outcomes other than trisomy 21 (T21),trisomy 18 (T18) and open neural tube defects (ONTD), and to provide local data for supporting evidence based clinical managements. Methods · A retrospective cohort study was performed in the women who received second trimester maternal serum screening in the International Peace Maternal and Child Health Hospital between 2012 and 2014, with naturally conceived singleton pregnancies. Obstetric outcomes were followed up by searching electronic medical records within the hospital. Abnormal level of marker was defined as a MOM value ≥ 99th (P99) or ≤ 1st percentile (P1) of the overall screened population. Incidence of an adverse obstetric outcome was compared between the groups with abnormal markers and the control with all markers in normal. Results · ① A total of 25616 pregnancies were included in this study, in which 4526 were identified as having various adverse obstetric outcomes. Among them 4143 pregnancies were with isolated and 383 pregnancies were with co-occurring two or more adverse outcomes. ② When compared to pregnancies with normal levels of all three serum markers, pregnancies with decreased AFP or decreased hCG did not show associations with any adverse obstetric outcomes. However, pregnancies with increased AFP, increased hCG or decreased uE3 were at increased risk for a variety of abnormal pregnancy outcome. In 18 pregnancies with an outcome of fetal chromosomal abnormalities other than T21 and T18, 9 presented with either increased AFP, increased hCG or decreased uE3, with relative risk ratios of 13.33、35.00 and 59.00, respectively. ③ The performance of those markers tended to be improved in a subset of adverse obstetric outcomes, including low birth weight

Analysis of AGG interspersion of the FMR1 gene in fragile X syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze (CGG)n repeats sequence and AGG interspersion correlated with unstable expansion of FMR1 gene in a general Chinese population.</p><p><b>METHODS</b>AmplideX FMR1 PCR Kit was used to amplify 380 X chromosomes from randomly selected 176 males and 102 females, 11 permutation carriers and 10 full mutation patients have served as controls. Results of capillary electrophoresis were analyzed with GeneMapper software Version 4.0. SPSS 11.0 software was used for statistical analysis.</p><p><b>RESULTS</b>The ratio of heterozygous females was 64.70%. The number of alleles in general males and females was 15 and 14, the classes of AGG pattern was 26 and 27, respectively. The range of alleles was between 17 to 45 CGG repeats in males and 21 to 44 CGG repeats in females, and 1 male was identified as gray zone carrier. The most frequent allele was 29 CGG repeats, which was followed by 30 and 36 repeats, while 28 CGG repeats were absent. The most common AGG pattern was 9A9A9, 99.21% of the population was detected with different forms and numbers of AGG interruption, and 6A interruption pattern was found in 10.02% samples especially in individuals with more CGG repeats. However, 57.58% of control samples had no AGG interruption, and none of the controls had 6A interruption pattern. No significant difference was observed in allele frequent distribution of (CGG)n repeats and AGG interspersion patterns between the males and females (P > 0.05), and AGGs was significantly different between general population and controls (P < 0.05).</p><p><b>CONCLUSION</b>AGGs and AGG pattern may have important roles in maintaining (CGG)n stability in general population of China, 9A9A6A9 may be a special pattern for preventing (CGG)n unstable expansion in Asian populations.</p>

Unbalanced subtelomic rearrangement involving 9q and 22q in a child with mental retardation and multiple congenital anomalies / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine genetic cause for a patient with development delay and multiple congenital anomalies.</p><p><b>METHODS</b>Routine karyotype analysis was performed for the patient and his parents. Array comparative genomic hybridization (array CGH) was performed for the patient to detect cryptic chromosome aberration.</p><p><b>RESULTS</b>Karyotype analysis revealed no obvious anomaly for the patient and his parents. Array CGH has detected a 2.8 Mb heterozygous deletion at 9q34.3 and an 8.1 Mb heterozygous duplication at 22q. Fluorescence in situ hybridization analysis of the patient revealed an unbalanced subtelomeric translocation 46, XY, der(9) t(9; 22) (q34.3; q13.2q13.33) mat, which has resulted in partial trisomy 22q and partial monosomy 9q. Clinical features of the patient included developmental delay, facial dysmorphism and multiple congenital anomalies. Upon subsequent pregnancy, FISH analysis revealed that the fetus has inherited the normal chromosomes 9 and 22 from his mother. Postnatal follow-up confirmed normal development milestone and physiques in the child.</p><p><b>CONCLUSION</b>An unbalanced translocation involving 9q and 22q has been found in a child featuring multiple congenital anomalies, which is due to a balanced translocation 9; 22 in his mother. Array CGH and FISH have also helped with discovery of subtelomeric rearrangement. Prenatal diagnosis of this aberration in subsequent pregnancies with FISH can prevent the recurrence of this disease.</p>

A pilot study on spinal muscular atrophy carrier screening in Shanghai region using real-time PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To develop a screening program for spinal muscular atrophy (SMA) carriers, and to assess the carrier frequency and detection rate in Shanghai region.</p><p><b>METHODS</b>Quantitative analysis of the SMN1 gene by real-time PCR was developed using specimens from 15 SMA patients and 76 SMA parents from 38 affected nuclear families. A pilot screening was carried out for 1741 asymptomatic pregnant women. Frequencies of SMN1 alleles were determined with the Hardy-Weinberg equilibrium.</p><p><b>RESULTS</b>Forty five out of the 1741 women were identified as SMA carriers by the presence of single copy of SMN1. The frequencies of no copy, 1 copy, 2 copy and 3 copy alleles were 1.37 U+00D7 10-2, 9.45 U+00D7 10-1, 2.80 U+00D7 10-2 and 1.27 U+00D7 10-2, respectively. The adjusted SMA carrier frequency was 1:35 with a detection rate of 94.49%. For those with a negative screening result, individuals with 3 copies carried a higher residual risk.</p><p><b>CONCLUSION</b>The incidence of SMA carriers in Shanghai region is similar with that in Caucasian populations. Carrier screening has high detection efficiency. An effort should be made to further distinguish SMN1 gene copy numbers for those with more than 2 copies, since accurate determination of 2 and 3 copy allele frequencies is essential for post-screening genetic consulting.</p>

Molecular and cytogenetic characterization of six 46, XX males due to translocations between the short arms of X and Y chromosomes / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To characterize molecular and cytogenetic abnormalities in six 46, XX males, and to investigate the clinical manifestations and underlying mechanisms in such patients.</p><p><b>METHODS</b>Clinical data of six XX male patients were collected. Karyotyping, multiple polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) were utilized to detect and locate the sex determining region (SRY) gene.</p><p><b>RESULTS</b>PCR and FISH showed that all patients were SRY-positive XX males. All patients have their SRY gene located at the tip of derivative X chromosomes, which have resulted from translocation between short arms of X and Y chromosomes. High resolution karyotyping at 550-750 band level has revealed that the translocation breakpoints were at Xp22.33 and Yp11.2 in three patients. In the remaining patients, the breakpoints were either at Xp22.32 and Yp11.31 or Xp22.31 and Yp11.2. The breakpoints at Xp22.32, Xp22.31 and Yp11.31 were rarely reported. Genotype-phenotype correlation analysis indicated that the clinical manifestations were age-specific. Four adult patients have come to clinical attention due to infertility, with typical features including azoospermia and testis dysgenesis, whereas poorly developed secondary sexual characteristics and short stature were main complaints of adolescence patients, and short stature was the sole symptom in a child patient.</p><p><b>CONCLUSION</b>Combined karyotyping, PCR and FISH are important for the analysis of XX males. Particularly, high resolution karyotyping is valuable for the refinement of chromosome breakpoints and detailed analysis of genotype-phenotype correlation.</p>

Clinical and genetic study of a case with Smith-Magenis syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the clinical feature and genetic diagnosis for Smith-Magenis syndrome (SMS).</p><p><b>METHOD</b>The clinical data, including craniofacial anomalies, physical and mental status were analyzed. Routine and high resolution G-banding was performed to analyze the karyotype of the patient and her parents, and array comparative genomic hybridization (array CGH) was used to detect small chromosome anomaly.</p><p><b>RESULT</b>A-two-year old girl was sent to our clinic for mental retardation and cardiac malformation. Some sleep problems were reported by parents, including difficulties falling asleep, shortened sleep cycles. She also had some neurobehavioral symptoms including hyperactivity and self-injurious behaviors head-banging. She had distinctive craniofacial features including low hairline, frontal bossing, a broad face, broad nasal bridge, a tented upper lip, prognathism, low-set ears and high-vaulted arch. She had moderate mental retardation. Cardiac findings included ventricular septal defect, atrial septal defect, overriding aorta and pulmonary hypertension. Primary ventriculomegaly was seen in magnetic resonance imaging (MRI). Routine karyotype analysis showed a karyotype of 46, XX. However, high resolution karyotype analysis showed a suspected partial deletion of the short arm of chromosome 17. Array comparative genomic hybridization (array CGH) finely mapped the deletion to a 3.8 Mb region on 17p11.2. The molecular karyotype was then ascertained as 46, XX.arr17p11.2(16543655-20374751)×1dn. The parents had normal karyotypes.</p><p><b>CONCLUSION</b>Smith-Magenis syndrome is a multisystem disorder characterized by developmental delay and mental retardation, distinctive craniofacial features, sleep disturbance and behavioral problems. Array comparative genomic hybridization (array CGH) finely mapped the deletion on 17p11.2.</p>

Two cases of partial trisomy 8p derived from paternal reciprocal translocation or maternal insertion translocation: clinical features and genetic abnormalities / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the origin of aberrant chromosomes involving the short arm of chromosome 8 in two mentally retarded children, and to correlate the karyotype with abnormal phenotype.</p><p><b>METHODS</b>Routine G-banding was performed to analyze the karyotypes of the two patients and their parents, and array comparative genomic hybridization (array CGH) was used for the first patient for fine mapping of the aberrant region.</p><p><b>RESULTS</b>The first patient presented with only mental retardation. The father had normal karyotype. The mother had an apparent insertion translocation involving chromosomes 8 and 3 [46, XX, inv ins (3; 8) (q25.3; p23.1p11.2)], the karyotype of the child was ascertained as 46, XX, der(3) inv ins (3; 8)(q25.3; p23.1p11.2). Array CGH finely mapped the duplication to 8p11.21-8p22, a 26.9 Mb region. The other patient presented with mental retardation, craniofacial defects, congenital heart disease and minor skeletal abnormality. The mother had normal karyotype. The father had an apparently balanced translocation involving chromosome 8p and 11q, the karyotype was 46, XY, t(8; 11)(p11.2; q25). The karyotype of the child was then ascertained as 46, XX, der(11)t(8; 11)(p11.2; q25).</p><p><b>CONCLUSION</b>These results suggested that partial trisomy 8p was primary cause for the phenotypic abnormalities of the two patients, whereas a mild phenotypic effect was observed in patient 1. Parental karyotype analysis could help define the aberrant type and recurrent risk evaluation. In contract to routine karyotype analysis, aberrant regions could be mapped by array CGH with higher resolution and accuracy.</p>

Optimization of polymerase chain reaction assay combined with capillary electrophoresis to detect the pre- and full mutation of FMR1 gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To develop an efficient, high resolution PCR assay suitable for detection of the (CGG)n repeats of the fragile X mental retardation 1 (FMR1) gene by optimizing the PCR system in combination with capillary electrophoresis.</p><p><b>METHODS</b>Three standard samples and twelve samples that were verified by Southern blot analysis including both male and female in the normal, pre- and full mutation range were used in this study to evaluate the enhanced PCR system. All amplicons were electrophoresed by agarose, polyacrylamide and capillary electrophoresis to compare the results.</p><p><b>RESULTS</b>The enhanced PCR assay developed in this study was able to detect a full mutation with (CGG)n being larger than 260 repeats in a male. An expanded pre-mutation allele with (CGG)n as large as 183 repeats in a female was also amplified. The capillary electrophoresis method used in this study was able to distinguish two alleles with 1 CGG repeat difference and the results were reproducible.</p><p><b>CONCLUSION</b>A high resolution PCR assay is developed, which is much more efficient than the general PCR systems. It is suitable for the clinical screening of FMR1 gene and will greatly reduce the number of Southern blot analysis needed in clinical application.</p>

A case with partial trisomy 7 (q34→qter) derived from a paternal reciprocal translocation t(7;14)(q34;q32) / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the origin of chromosomal aberrants in a mentally retarded children, and to correlate the karyotype with phenotype.</p><p><b>METHODS</b>Routine G-banding were performed to analyze the karyotype of the patient and her parents, and array comparative genomic hybridization (array CGH) were used for finely mapping the aberrant regions.</p><p><b>RESULTS</b>The mother had a normal karyotype. The father had an apparently balanced translocation involving chromosome 7q and 14q, the karyotype was 46, XX, t(7;14) (q34;q32), the karyotype of the child was then ascertained as 46, XX, der(14) t(7;14) (q34;q32.33) pat. Array CGH finely mapped the duplication to 7q34-qter, a 17.09 Mb region, and a very small associated deletion of distal chromosome 14 to 14q32.33-qter, a 2.27 Mb region. The patient presented some frequently seen features in partial trisomy 7q cases such as mental retardation, low birth weight, small nose, cleft palate, low-set ears and short neck.</p><p><b>CONCLUSION</b>This result suggested that partial trisomy 7q exert mainly phenotypic effect on the patient. Parental karyotype analysis could help define the aberrant type.</p>

Clinical application of multiplex ligation-dependent probe amplification for the detection exonic copy number alterations in the Dystrophin gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To clarify advantages and disadvantages of using multiplex ligation-dependent probe amplification (MLPA) for detecting exonic deletions and duplications of the Dystrophin gene, and to explore the appropriate management for single-exon abnormality detected by MLPA.</p><p><b>METHODS</b>MLPA were performed to detect exonic copy number changes in 70 Duchenne/Becker muscular dystrophy (DMD/BMD) patients diagnosed by clinical and histological findings. PCR, DNA sequencing and real-time PCR were applied to the samples in which MLPA indicated single-exon deletion or duplication.</p><p><b>RESULTS</b>Of all 70 patients, MLPA detected exonic deletions in 42 (60%), including 12 with single-exon deletion and one with ambiguous single-exon deletion. Exon duplications were found in 7 patients (10%), among which two were single-exon duplication. 21 patients showed normal results (30%). For the 12 patients with single-exon deletion, MLPA results were confirmed by PCR in 11. In one patient, a deletion of two nucleotides (c.4470-4471delAA) was found by sequencing. A novel two-nucleotide deletion (c.4746-4747delCT) was identified in the patient with the ambiguous single-exon deletion. For the two patients showed single-exon duplication, MLPA results were confirmed by real-time PCR.</p><p><b>CONCLUSION</b>MLPA should be the first choice in detecting Dystrophin gene exon deletions and duplications. Single-exon deletion/duplication resulted from MLPA should be further evaluated by other methods.</p>

Genetic tests and clinical re-evaluation of 85 children with suspected spinal muscular atrophy / 中华儿科杂志

<p><b>OBJECTIVE</b>Spinal muscular atrophy (SMA), characterized by degeneration of the anterior horn cells in the spinal cord and symmetric proximal muscle weakness, is the most common autosomal recessive neuromuscular disease in infants and children. In Caucasian population, about 95% of clinically typical patients lack both copies of the telomeric survival motor neuron gene (SMN 1). However, the detection rate of the homozygous absence in Chinese patients is still controversial, which may lead to reduced confidence in the SMA genetic testing in clinical practice. The purpose of the current study was to determine the frequency of homozygous deletions of SMN 1 in Chinese patients, to evaluate the significance of the SMN 1 homozygous deletion assay in clinical applications, and the impact of the clinical re-visit followed by the genetic testing.</p><p><b>METHODS</b>Totally 85 patients initially suspected of SMA were referred for SMA genetic testing. A polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) assay was used to detect the homozygous absence of SMN 1. Clinical re-visit was performed by the pediatric neurology specialists according to the international SMA diagnostic criteria, and histological examinations were carried out when they were necessary.</p><p><b>RESULTS</b>Absence of both copies of SMN 1 exon 7 were found in 57 (67%) of the 85 patients, and 28 patients (33%) had at least one copy. For the 28 patients with negative results, 19 were followed up by the pediatric neurologists. The clinical diagnosis of SMA could be excluded in 15 patients, but retained in the other 4 patients after the clinical re-evaluation and histological examinations. Thus, approximately 95% of the patients with clinically typical SMA in our cohort lacked both copies of SMN 1. Homozygous deletions of SMN 1 were detected in 96% (22/23), 93% (28/30) and 100% (7/7) of the patients with SMA type I, type II and type III, respectively. There was no significant difference in the deletion frequency among the subtypes.</p><p><b>CONCLUSIONS</b>The frequency of homozygous deletions of SMN 1 in this series of Chinese SMA patients was about 95%, which is similar to that reported in Caucasian population. The genetic test of homozygous deletions of SMN 1 should be considered as the first line test for the Chinese patients suspected of SMA. The clinical re-visit and re-evaluation which is essential in clinical diagnosis, genetic counseling and medical management, should be routinely performed after the genetic testing.</p>

Cytogenetic and molecular genetic study of a case with 8p inverted duplication deletion syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To define the origin and the precise location of the aberrant fragments on the short arm of the chromosome 8 in a mentally retarded boy, and to understand the mechanism, the characteristic clinical features and the recurrent risk associated with this abnormality.</p><p><b>METHODS</b>High-resolution chromosomal banding was performed to analyze the karyotype of the patient and his parents, array comparative genomic hybridization (array-CGH) was employed to investigate the precise location of the aberrant fragments, and quantitative real-time PCR was used to confirm the results.</p><p><b>RESULTS</b>The rearranged chromosome 8 in the patient was inverted and duplicated for region 8p11.2-p23.1, and deleted for region 8p23.1-pter. In between, a 5.70 Mb single copy region was present, which was delimited by the two olfactory receptor (OR) gene clusters.</p><p><b>CONCLUSION</b>This is a case of classic inv dup del(8p) syndrome, which is characterized by severe mental retardation, brain malformation and specific facial dysmorphism, and is induced by non-allelic homologous recombination (NAHR) between the OR genes on 8p23.1. Prenatal diagnosis should be performed to monitor the recurrent risk of inv dup del(8p), as well as the other three harmful consequences resulted from the same NAHR mechanism. To the best of our knowledge, this is the first case of inverted duplicated 8p syndrome identified in Mainland China.</p>

Successful prenatal diagnosis following elimination of maternal cell contamination in a family with recessive dystrophic epidermolysis bullosa / 中华皮肤科杂志

Objective To perform a DNA-based prenatal diagnosis in a family with recessive dys-trophic epidermolysis bullosa, and to develop a strategy to eliminate matemal cell contamination in arnniotic fluid samples. Methods Amniocentesis was carried out at gestation week 16, amniotic fluid culture was used to separate fetal cells from maternal blood cells. Peripheral blood was obtained from the proband, and her parents. Genomic DNA was extracted from peripheral blood and aminotic cells. Subsequently, PCR and direct sequencing were performed to detect pathogenic mutations in the COL7A1 gone. Karyotype analysis was used to confirm paternal information in amniotic fluid. Linkage analysis between micro-satellite markers was performed to confirm the fetal genotype. Resulta Centrifugation showed visible contamination of aminotic cells by blood cells. Direct sequencing revealed that the proband was a carrier of both maternal mutation, R525X in exon 12, and paternal mutation, R2610X in exon 105, while the fetus only carried the maternal mutation, R525X. The second direct sequencing and hapiotype analysis after elimination of mater-nal blood cells by amniotic fluid culture confirmed that the fetus was a carrier of maternal mutation with nor-real phenotype. The pregnancy continued and a clinically unaffected girl was born at gestation week 40.Conclusion The accuracy of DNA-based prenatal diagnosis could be improved by the combination of direct sequencing, amniotic fluid culture, karyotype analysis and linkage analysis, etc.

The utility of Liebowitz Social Anxiety Scale in the patients with social anxiety disorder in Chinese / 中国神经精神疾病杂志

Background During the past two decades, a number of rating scales were developed to facilitate diagnosis and assessment of subjects with social anxiety disorder. One of the most commonly used scales for the assessment of social anxiety disorder is the Liebowitz social anxiety scale (LSAS). The LSAS is widely used in epidemiologic investigations and clinical researches,and its assessment in the pharmacotherapy efficacy for social anxiety disorder is superior to any other scale. So we designed this study to explore the validity and reliability of the LSAS in Chinese patients with social anxiety disorder and normal control, and to find the difference of the scores between the patients self -report version and clinician-administered version. Methods Fifty five patients meeting the DSM-Ⅳ diagnostic criteria for social anxiety disorder and 168 normal controls who were screened from 222 college students were rated by LSAS, social phobia scale and self-made General Information Forms. Results The Cronbath α of LSAS for the patients and the normal controls was 0.83 and 0.77, respectively. The 4-week test-retest reliability for total scores and its factors scores of LSAS in 31 normal controls were ranging from 0.68 to 0.79. The ROC area under curve value in discriminating the patients from normal controls was 0.87±0.03; the total score of 35 was considered to be the best cut-off score for LSAS, then its sensitivity was 0.77 and its specificity was 0.81; and no significant difference between the self-report version and clinician-administered version. Conclusions The LSAS is good in internal consistency and test-retest reliability, and has high sensitivity and specificity in discriminating the patients and the controls. There is no significant difference in the total score and each factor scores of LSAS between self-report version and clinician-administered version.