Aeromonas immobilized on chitosan for treating high-oil wastewater from kitchens / 生物工程学报

To effectively solve the serious impact of high oil in the kitchen wastewater on the downstream treatment process, an excellent oil-degrading strain Aeromonas allosaccarophila CY-01 was immobilized to prepare Chitosan-Aeromonas pellets (CH-CY01) by using chitosan as a carrier. Oil degradation condition and efficiency of CH-CY01 pellets were assessed. The growth of immobilized CH-CY01 was almost unaffected, and the maximum degradation rate of soybean oil was 89.7%. Especially at 0.5% NaCl concentration, oil degradation efficiency of CH-CY01 was increased by 20% compared with free cells. In the presence of a surfactant (sodium dodecylbenzene sulfonate) at 1 mg/L, the degradation efficiency of oil by CH-CY01 was increased by 40%. Moreover, using the high-oil catering wastewater as the substrate, more than 80% of the solid oil was degraded with 1% (V/V) CH-CY01 pellets treatment for 7 days, significantly higher than that of free cells. In summary, immobilized CH-CY01 significantly improved the efficiency of oil degradation.

Diagnosis of Spinal Muscular Atrophy: A Simple Method for Quantifying the Relative Amount of Survival Motor Neuron Gene 1/2 Using Sanger DNA Sequencing / 中华医学杂志(英文版)

Background@#Spinal muscular atrophy (SMA) is caused by homozygous deletion or compound heterozygous mutation of survival motor neuron gene 1 (SMN1), which is the key to diagnose SMA. The study was to establish and evaluate a new diagnostic method for SMA.@*Methods@#A total of 1494 children suspected with SMA were enrolled in this study. Traditional strategy, including multiplexed ligation-dependent probe amplification (MLPA) and TA cloning, was used in 1364 suspected SMA children from 2003 to 2014, and the 130 suspected SMA children were tested by a new strategy from 2015 to 2016, who were also verified by MLPA combined with TA cloning. The SMN1 and SMN2 were simultaneously amplified by polymerase chain reaction using the same primers. Mutation Surveyor software was used to detect and quantify the SMN1 variants by calculating allelic proportions in Sanger sequencing. Finally, turnaround time and cost of these two strategies were compared.@*Results@#Among 1364 suspected SMA children, 576 children had SMN1 homozygous deletion and 27 children had SMN1 compound heterozygous mutation. Among the 130 cases, 59 had SMN1 homozygous deletion and 8 had heterozygous deletion: the SMN1-specific peak proportion on exon 7 was 34.6 ± 1.0% and 25.5 ± 0.5%, representing SMN1:SMN2 to be 1:2 and 1:3, respectively. Moreover, five variations, including p.Ser8Lysfs *23 (in two cases), p.Leu228*, p.Pro218Hisfs *26, p.Ser143Phefs*5, and p.Tyr276His, were detected in 6/8 cases with heterozygous deletion, the mutant allele proportion was 31.9%, 23.9%, 37.6%, 32.8%, 24.5%, and 23.6%, which was similar to that of the SMN1-specific site on exon 7, suggesting that those subtle mutations were located in SMN1. All these results were consistent with MLPA and TA cloning. The turnaround times of two strategies were 7.5 h and 266.5 h, respectively. Cost of a new strategy was only 28.5% of the traditional strategy.@*Conclusion@#Sanger sequencing combined with Mutation Surveyor analysis has potential application in SMA diagnosis.

Status of healthcare-associated infection management in 42 Chinesemedi-cine hospitals in Fujian Province / 中国感染控制杂志

Objective To understand the status of healthcare-associated infection(HAI)management in traditional Chinese medicine hospitals as well as integrated traditional Chinese and Western medicine hospitals in Fujian Pro-vince,analyze the existing problems and weak links,and put forward corresponding improvement measures.Methods A questionnaire was designed through literature and expert consultation,from March to April 2016,42 secondary and above traditional Chinese medicine hospitals as well as integrated traditional Chinese and Western medicine hos-pitals in 8 cities of Fujian Province were conducted on-site investigation,data were analyzed.Results A total of 42 hospitals participated in the investigation,92.86% were traditional Chinese medicine hospitals,7.14% were inte-grated traditional Chinese and Western medicine hospitals;all hospitals set up HAI management committees and HAI management groups of clinical departments,there were 100 HAI management professionals(66 were full-time,34 were part-time),nursing staff accounted for 63.00%,junior college and undergraduate personnel accoun-ted for 84.00%,staff with intermediate and senior professional titles accounted for 79.00%.There were significant differences in academic disciplines and education levels among administrators in secondary and tertiary hospitals(P<0.05). All hospitals carried out HAI case surveillance,only 2.38% achieved HAI informational software monito-ring,83.33% carried out comprehensive and targeted monitoring,42.86%,71.43%,and 80.95% of hospitals car-ried out targeted monitoring on multidrug-resistant organisms,surgical site infection,and intensive care unit respec-tively.Conclusion The environment of majority of Chinese medicine hospitals in Fujian Province improved signifi-cantly,organizations of HAI management is rational,staffing and quality of HAI management personnel is imbal-anced,HAI monitoring is still at preliminary stage,lack information management,HAI management in key depart-ments is not optimistic.

Arthroscopic debridement and meniscectomy in treatment of meniscus injury combined with osteoarthritis / 中国内镜杂志

Objective To investigate the effect of arthroscopic debridement and meniscectomy in treatment of moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage. Methods 156 cases diagnosed with moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage were collected from October 2011 to October 2014. Lysholm knee score and preoperative examinations such as anteroposterior, lateral, axial radiographs, the standing full leg length X-ray film and MRI scan of the knee were recommended to definitively understand the osteoarthritis staging and meniscus injury grading. All patients were treated with arthroscopic debridement and meniscectomy. After operation, physical rehabilitation exercises and regular clinical follow-up were carried out as planned. The Lysholm knee score data from preoperation and terminal follow-up was statistical analyzed. Results No patient experienced any perioperative and postoperative complications. Statistical analysis showed that the Lysholm knee score of postoperation was significantly higher than that of preoperation [(87.3 ± 7.9) vs (67.5 ± 4.9), P < 0.05). Conclusion Arthroscopic debridement and meniscectomy in treatment of moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage, gains beneficial effects for its minimal invasion and quick recovery.

Multislice CT in diagnosis of associated carpal bone fractures in distal radial fractures / 中华全科医师杂志

Clinical data and radiological findings of 78 patients with distal radial fractures,who underwent plain X-ray film and muhislice CT (MSCT) examinations,were retrospectively analyzed.Twenty nine associated carpal bone factures were detected on X-ray film in 21 cases;while 47 associated carpal bone fractures were detected on MSCT in 29 cases (P ＜ 0.05).The missed diagnosis rate of X-ray was 38％.Results indicate that MSCT can significantly improve the detect rate,which should be recommended for diagnosis of associated carpal bone fractures in distal radial fractures.

Detection of homozygous deletions in spinal muscular atrophy with genomic DNA sequencing / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect homozygous deletions of survival motor neuron (SMN) gene with genomic DNA sequencing, and to assess the value of genetic testing for the diagnosis of spinal muscular atrophy (SMA).</p><p><b>METHODS</b>Polymerase chain reaction (PCR) was used for amplifying SMN gene in 100 SMA patients and 110 controls. Four different bases (g.31957, g.32006, g.32154 and g.32269) between SMN1 and SMN2 within the amplified segments were identified with genomic DNA sequencing. Homozygous deletion of SMN1 or SMN2 was determined by the presence or absence of base peaks at such four sites. Multiplex ligation-dependent probe amplification (MLPA) was carried out to confirm the results of genomic DNA sequencing.</p><p><b>RESULTS</b>In the 100 SMA samples, only SMN2 specific base peaks were detected at the four sites, for which the copy numbers of SMN1 and SMN2 was 0:2 or 0:3, suggesting homozygous deletion of SMN1 gene. By contrast, only SMN1 specific base peaks were detected in 5 samples, for which the ratio of SMN1:SMN2 was 2:0, indicating homozygous deletion of SMN2. At four different sites, SMN1/SMN2 heterozygous peaks were detected in the remaining 105 samples, for which SMN1:SMN2was 2:2, suggesting non-deletion of SMN1 or SMN2. The results of sequencing were consistent with those of MLPA.</p><p><b>CONCLUSION</b>Genomic DNA sequencing is a rapid, accurate and economic method for the diagnosis of homozygous deletion of SMA.</p>

Differentiated miRNA expression and validation of signaling pathways in apoE gene knockout mice by cross-verification microarray platform

The microRNA (miRNA) regulation mechanisms associated with atherosclerosis are largely undocumented. Specific selection and efficient validation of miRNA regulation pathways involved in atherosclerosis development may be better assessed by contemporary microarray platforms applying cross-verification methodology. A screening platform was established using both miRNA and genomic microarrays. Microarray analysis was then simultaneously performed on pooled atherosclerotic aortic tissues from 10 Apolipoprotein E (apoE) knockout mice (apoE-/-) and 10 healthy C57BL/6 (B6) mice. Differentiated miRNAs were screened and cross-verified against an mRNA screen database to explore integrative mRNA-miRNA regulation. Gene set enrichment analysis was conducted to describe the potential pathways regulated by these mRNA-miRNA interactions. High-throughput data analysis of miRNA and genomic microarrays of knockout and healthy control mice revealed 75 differentially expressed miRNAs in apoE-/- mice at a threshold value of 2. The six miRNAs with the greatest differentiation expression were confirmed by real-time quantitative reverse-transcription PCR (qRT-PCR) in atherosclerotic tissues. Significantly enriched pathways, such as the type 2 diabetes mellitus pathway, were observed by a gene-set enrichment analysis. The enriched molecular pathways were confirmed through qRT-PCR evaluation by observing the presence of suppressor of cytokine signaling 3 (SOCS3) and SOCS3-related miRNAs, miR-30a, miR-30e and miR-19b. Cross-verified high-throughput microarrays are optimally accurate and effective screening methods for miRNA regulation profiles associated with atherosclerosis. The identified SOCS3 pathway is a potentially valuable target for future development of targeted miRNA therapies to control atherosclerosis development and progression.

Analysis of chromosomal karyotypes, age and height on diagnosis of 232 patients with Turner syndrome / 中华实用儿科临床杂志

Objective To study the distribution characteristics of chromosomal karyotypes,age and height on diagnosis of patients with Turner syndrome(TS).Methods Karyotypes analysis was performed in 1015 girls with short stature who visited the hospital in recent 27 years.And statistical analysis of the chromosomal karyotypes,age and height on the diagnosis of patients with TS were performed by using the SPSS 11.0 software.Results 1.Two hundred and thirty-two (22.9％) patients were diagnosed with TS.2.A total of 26 kinds of abnormal karyotypes were detected in 232 patients with TS and the 45,X (32.4％,75/232 cases) and 46,Xi (Xq) (16.4％,38/232 cases) were the more common karyotypes.In addition,there were 111 cases(47.8％) with 21 kinds of mosaicisms which were related to the number and/or structural anomalies of one X chromosome.3.The median(range) age on the diagnosis was (10.90 ±4.55) years old.TS was mainly discovered during adolescence.The diagnosis rates at different ages were as follows:＜2 years 15 cases(6.4％),2-＜7 years 35 cases(15.1％),7-＜ 12 years 93 cases(40.1％) and 12-＜ 18 years 89 cases(38.4％).4.The median height Z score was-3.60 ± 1.20.There was a significant negative linear correlation between age and Z score of height on the diagnosis of TS (r =-0.613,P ＜ 0.000).5.Eighty-six point five percent (64/74 cases)TS patients with age over 13 years presented pubertal delay at diagnosis,which occurred only in 27.2 ％(22/81 cases) of the control group,and there existed a significant difference(OR =16.297,P =0.000).Conclusions 1.Chromosomal karyotypes in TS are in diverse ness.2.In TS patients,as the age increases,delay of height and pubertal development could be more obvious to observe and more characteristic manifestation of pubertal development would appear.3.The diagnosis of TS was later in children in our cotmtry than that in the developed countries.The analysis of chromosomal karyotypes should be detected earlier in the girls with unexplained short stature,height below-2 Z score of the mean age or the height below the specific lower limit of the peers,so as to benefit the diagnose of TS earlier.

MRI study on predicting the collapse of avascular necrosis of the femoral head / 中华放射学杂志

Objective To study the risk factors of MRI for the prediction of collapse in patients with avascular necrosis of the femoral head.Methods Twenty-two patients (39 hips) diagnosed avascular necrosis of femoral head by MR were enrolled in our study.The following MR appearances were evaluated:bone marrow edema,joint fluids,signal intensity and location of the lesion.The volume and surface area of the necrosis zone were calculated.The time of follow-up was 18-84 months (median,25 months).Logistic regression analysis was used to predict the risk factors by SPSS 13.0.The maximum value of Youden index was selected as the critical point to predict the collapse of femoral head and to define the sensitivity,specificity and accuracy.Results In the 39 hips with femoral head necrosis,21 hips had collapse.Bilateral collapse occurred in 5 cases.In 25 hips with the necrosis surface larger than 25％,collapse occurred in 21 (84％); In 8 hips with the volume of femoral head necrosis larger than 30％,collapse occurred in all cases; 1n 33 hips with the necrosis locating at the superolateral quadrant,collapse occurred in 21 (63.6％); In 22 hips with necrotic areas showing heterogeneous signal intensity,collapse occurred in 18(81.8％) ;In 25 hips with large amount of joint effusion,collapse occurred in 16 (64％) ;in 18 hips with bone marrow edema,collapse occurred in 13 (65％).Joint fluid,heterogeneous signal intensity and lesions in the superolateral quadrant,volume ratio,and area ratio were the high risk factors,while bone marrow edema was a relatively low risk factor.The area under ROC curves for area ratio of NASA was greater than that for volume ratio (0.987 vs 0.902).When the critical value for area ratio was 26.7％,the true positive rate was 95.2％,true negative rate was 94.4％,and Youden's index was 0.896.Conclusions The collapse of necrosis of femoral head may result from many factors.The femoral head was easy to collapse when it had large enough area of necrosis and mixed signal intensity,a large amount of joint effusion,bone marrow edema,and superolateral quadrant location.The critical value for area ratio to predict the collapse of femoral head was about 26.7％.The area ratio is more accurate than volume ratio in predicting the collapse of necrosis of femoral head.

Compound mutations (R237X and L375P) in the fumarylacetoacetate hydrolase gene causing tyrosinemia type I in a Chinese patient / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mutations in fumarylacetoacetate hydrolase (FAH) gene can lead to tyrosinemia type 1 (HT1), a relatively rare autosomal recessive disorder. To date, no molecular genetic defects of HT1 in China have been described. We investigated a Chinese family with a HT1 child to identify mutations in FAH.</p><p><b>METHODS</b>DNA sequencing was used for mutations screening in FAH gene. Real-time polymerase chain reaction (PCR) was performed to determine the FAH gene expression level. To confirm the presence of degradation by the nonsense-mediated mRNA decay pathway (NMD), the fragments containing R237X mutations were analyzed by primer introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and cDNA sequencing. Finally, the effects of the mutations reported in this study were predicted by online softwares.</p><p><b>RESULTS</b>A boy aged 3 years and 8 months was diagnosed clinically with HT1 based on his manifestations and biochemical abnormalities. Screening of FAH gene revealed two heterozygous mutations R237X and L375P transmitted from his mother and father respectively. In this pedigree, the amount of FAH mRNA relative to a healthy control was 0.44 for the patient, 0.77 for his mother and 1.07 for his father. Moreover, both PIRA-PCR and cDNA sequencing showed significant reduction of the FAH mRNA with R237X nonsense mutation. The missense mutation of L375P was not reported previously and prediction software showed that this mutation decreased the stability of protein structure and affected protein function.</p><p><b>CONCLUSIONS</b>This is the first case of HT1 analyzed by molecular genetics in China. The R237X mutation in FAH down- regulates the FAH gene expression, and the L375P mutation perhaps interrupts the secondary structure of FAH protein.</p>

Limitation of PCR-RFLP method for the detection of genetic mutations in spinal muscular atrophy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the applicability and limitation of PCR-restriction fragment length polymorphism (PCR-RFLP) method for genetic diagnosis of spinal muscular atrophy (SMA).</p><p><b>METHODS</b>PCR-RFLP was applied to detect potential deletion in exons 7 and 8 of SMN1 gene in 935 suspected cases with SMA. Multiplex ligation-dependent probe amplification(MLPA) was carried out to analyze dosage alteration of SMN1 gene in 339 of such cases. To confirm the accuracy of PCR-RFLP method for homozygous and heterozygous deletions detection, the consistency of PCR-RFLP and MLPA results were assessed with a Pearson Chi-square test.</p><p><b>RESULTS</b>Homozygous deletion of exon 7 of SMN1 was detected in 590 suspected cases. The rate of diagnosis was therefore 63.1% (590/935). For the 339 suspected cases, PCR-RFLP and MLPA respectively identified 194 and 196 homozygous deletions in the exon 7 of SMN1 gene, suggesting a good consistency (98.9%)(Chi-square = 0.2, P = 0.88). However, only 4 of 339 cases was found to carry a heterozygous deletion of SMN1 exon 7 by PCR-RFLP, in contrast with 17 detected by MLPA. The consistency only reached 23.5%, for which statistical significance was detected (Chi-square = 8.29, P< 0.01).</p><p><b>CONCLUSION</b>Although PCR-RFLP is a simple, specific and efficient method for SMA diagnosis, it has obvious limitation for the diagnosis of 5%-10% SMA patients who have carried a compound heterozygous mutation.</p>

Compound heterozygous mutation in two unrelated cases of Chinese spinal muscular atrophy patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Infantile proximal spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. Approximately 90% - 95% cases of SMA result from homozygous deletion of survival motor neuron gene 1 (SMN1) and 5% cases are caused by compound heterozygous mutation (a SMN1 deletion on one allele and a subtle mutation on the other allele).</p><p><b>METHODS</b>In this research, two unrelated patients were clinically diagnosed according to the criteria of proximal SMA. Genetic diagnosis was performed to detect the homozygous deletion of exon 7 of SMN1 by PCR-restriction fragment length polymorphism (RFLP) and genomic sequencing. Multiplex ligation-dependent probe amplification (MLPA) analysis was carried out to measure copy numbers of SMN1, SMN2 and neuronal apoptosis inhibitor protein (NAIP) in the patients. Further sequencing of SMN1 allele-specific PCR (AS-PCR) and SMN1 clones were also performed to analyze the point mutation of SMN1 gene. Additionally, the pedigree analysis of these two families was carried out to identify the transmission of the mutation.</p><p><b>RESULTS</b>The inconsistent results using PCR-RFLP and genomic sequencing showed homozygous deletion of exon 7 of SMN1 and heterozygous deletion accompanied with a suspicious mutation in SMN1 gene, respectively. MLPA analysis of these two cases exhibited one SMN1 copy deletion. One identical c.863G > T (p.Arg288Met) mutation was found in two cases by sequencing the SMN1 clones, which confirmed that both cases were SMA compound heterozygotes. One case showed partial conversion to form hybrid SMN (SMN2 I7/SMN1 E8) identified by clones sequencing and another case carrying 3 SMN2 implied complete conversion from SMN1 to SMN2.</p><p><b>CONCLUSION</b>p.Arg288Met is more a disease-causing mutation than a polymorphism variation, and children with this mutation may have more severe phenotypes.</p>

Point mutation analysis of SMN1 gene in patients with spinal muscular atrophy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the point mutations in survival motor neuron gene 1 SMN1 gene and confirm the existence of compound heterozygous mutations in Chinese patients with spinal muscular atrophy (SMA).</p><p><b>METHODS</b>Three unrelated patients were diagnosed and clinically typed according to the criteria of proximal SMA established by the International SMA Consortium. Multiplex ligation-dependent probe amplification (MLPA) analysis was carried out to measure the copy numbers of SMN1, SMN2 and neuronal apoptosis inhibitory protein gene (NAIP)in the patients. The point mutation analysis of SMN1 gene was performed by reversed transcript-polymerase chain reaction (RT-PCR) and cloning sequencing. The MLPA assay and point mutation analysis were also performed in the family members to confirm the transmission of the mutations.</p><p><b>RESULTS</b>Two point mutations were identified in the present study, i.e., the p.Leu228X in one patient and p.Arg288Met in two patients. The mutation p.Arg288Met was first reported in Chinese and p.Leu228X was first reported in Mainland Chinese. The case carrying p.Leu228X mutation was diagnosed as SMA I with 2 copies of SMN2, and the cases with p.Arg288Met were diagnosed as SMA I and SMA II , respectively, with 3 copies of SMN2 gene.</p><p><b>CONCLUSION</b>The mutations p.Leu228X and p.Arg288Met caused severe clinical phenotypes, SMA I or SMA II. This study suggested that the compound heterozygous mutations of SMN1 existed in Chinese SMA patients, which was rarely reported previously in Chinese. It was necessary to detect the point mutation in SMN1 for genetic diagnosis of those patients with heterozygous deletion of SMN1, which would be beneficial to prenatal diagnosis and genetic counseling in these families.</p>

Analysis of survival motor neuron gene conversion in patients with spinal muscular atrophy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the type and frequency of gene conversion from SMN1 to SMN2 in Chinese patients affected with spinal muscular atrophy (SMA), and to explore the relationship between gene conversion and clinical phenotype.</p><p><b>METHODS</b>Non-homozygous deletion of SMN1 gene exon 8 was screened among 417 patients with SMN1 exon 7 homozygous deletions. To analyze and verify the types of gene conversion, genomic DNA sequencing, multiplex ligation-dependent probe amplification (MLPA), and gene subcloning and sequencing were carried out.</p><p><b>RESULTS</b>Thirty-one patients (7.4% of all) with non-homozygous deletions of SMN1 exon 8 were detected. Through series of experiments, the fusion genes SMN1/SMN2 in all cases were delineated. Five types of gene conversions were identified, which included SMN2-I7b/SMN1 E8, SMN2-I7a/SMN1 I7b, SMN2-E7/SMN1 I7a, SMN1 I6/SMN2 E7/SMN1 I7a and SMN2-E7/SMN1 I7a/SMN2 I7b. Such conversions were found in the type I-III patients. For 10 patients with type I-III SMA and 3 copies of SMN2 gene produced by conversion, the average survival age was 5 year and 4 months.</p><p><b>CONCLUSION</b>Partial conversions of SMN1 gene have been found among Chinese SMA patients. The type of conversion and frequency seem to be different from those of other races. Gene conversion to some extent may impact on survival time and rate of SMA patients, especially type I SMA.</p>

A novel missense mutation of the ubiquitin protein ligase E3A gene in a patient with Angelman syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Angelman syndrome (AS) is a neurogenetic disorder caused by an expression defect of the maternally inherited copy of ubiquitin protein ligase E3A (UBE3A) gene from chromosome 15. Although the most common genetic defects include maternal deletions of chromosome 15q11-13, paternal uniparental disomy and imprinting defect, mutations in the UBE3A gene have been identified in approximately 10% of AS patients.</p><p><b>METHODS</b>A Chinese girl of 28 months presented clinical manifestation of AS. Genetic diagnosis and molecular genetic defects were studied by methylation-specific PCR (MS-PCR) and linkage analysis by short tandem repeat (STR). We further performed sequence analysis of all the coding exons and flanking sequences of the UBE3A gene. The novel mutation screening was also performed in 100 unrelated healthy individuals to exclude the possibility of identifying a polymorphism variation.</p><p><b>RESULTS</b>The MS-PCR analysis of the patient showed biparental inheritance of chromosome 15 with a normal methylation pattern in the 15q11-q13 region. And STR analysis revealed that the patient also inherited biparental alleles for six microsatellites. A novel mutation, cDNA1199 C> A (p.P400H), in exon 9 of the maternal UBE3A gene, was identified in the patient. Meanwhile, the mutation was observed in the patient's mother who had a normal phenotype.</p><p><b>CONCLUSIONS</b>It is necessary to perform the UBE3A gene mutation analysis in non-deletion/non-UPD/non-ID patients with AS. The clinical picture of the patient is concordant with that observed in previously reported AS patients with UBE3A mutation.</p>

Mutation analysis of SMN1 gene in patients with spinal muscular atrophy / 中华儿科杂志

<p><b>OBJECTIVE</b>Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder. About 80% - 90% of SMA patients are missing both copies of SMN1, and 5% - 10% of patients are compound heterozygotes. In the present study, we aimed to analyze survival motor neuron 1 (SMN1) gene mutation in three patients with spinal muscular atrophy and their families to explore the effect of mutation on SMN protein function and the relationship between mutation and clinical phenotype.</p><p><b>METHOD</b>According to the international criterion, all patients were diagnosed by a neurologist. Patient 1 is a 5 years old boy with SMA type II. Patient 2, female, 2.5 years old, was SMA type II. Patient 3, female, 9 years old, was SMA type III. The brother of patient 3 was SMA type II, too. The age at last examination was 14 years. Genomic DNA was extracted from peripheral blood leukocytes by using standard phenol/chloroform method and total RNA was extracted from whole blood with QIAamp RNA Blood Mini Kit. PCR/RFLP was used to detect the homozygosis deletion of the SMN1 exon 7, and multiplex ligation-dependent probe amplification (MLPA) were performed to analyze the gene dosage of SMN1 and SMN2 for each patient and his/her family members; reverse transcriptase (RT)-PCR and clone sequencing were conducted for identifying the point mutation of SMN1 in three patients. The sequencing of genomic DNA and MLPA were carried out in the 3 families members to confirm the transition of mutation.</p><p><b>RESULT</b>No homozygous deletion of the SMN1 exon 7 was observed in any member of the 3 families. Case 1 and case 2 had one SMN1 copy compound with c.400G > A (p.Glu134Lys) mutation on it and SMN2 was two copies, respectively. Case 3 and her brother also had one copy of SMN1 and two copies of SMN2, and a mutation c.689C > T (p.Ser230Leu) occurred on the retained SMN1. All point mutations were from their fathers and deletion come from their mothers for SMN1 gene.</p><p><b>CONCLUSION</b>In this work, p.Glu134Lys and p.Ser230Leu mutations were identified in three unrelated families and p.Glu134Lys from two patients was first discovered in Chinese SMA. The p.Glu134Lys mutation within the SMN Tudor domain prevents the binding of SMN and Sm. The fact that p.Ser230Leu results in a polar amino acid substituted for non-polar amino acid possibly affects the structure of SMN and then damages its function. SMN1 point mutation analysis is not only advantageous to the diagnosis of those patients with heterozygous deletion of SMN1, but will be beneficial to the prenatal diagnosis and genetic counseling for their families.</p>

Genetic and clinical study on 17 cases of Angelman syndrome with deletion of 15q11-13 / 中华儿科杂志

<p><b>OBJECTIVE</b>Angelman syndrome (AS) is a neurodevelopmental genetic disorder that maps to 15q11-13. The primary phenotypes are attributable to loss of expression of imprinted UBE3A gene within this region which can arise by means of a number of mechanisms. The purpose of this study was to make a genetic diagnosis and to analyze the clinical features in suspected patients with AS.</p><p><b>METHOD</b>A total of 17 cases were diagnosed clinically as AS including 7 males and 10 females. The age at the time of diagnosis ranged from 8 months to 5 years. Genetic diagnosis was made by methylation-specific PCR (MS-PCR), linkage analysis by short tandem repeat (STR) and chromosome karyotype analysis. According to the international diagnostic criteria of AS, the related characteristic clinical features of the AS patients with deletion of 15q11-13 were analyzed and summarized.</p><p><b>RESULT</b>Deletion of 15q11-13 was confirmed by genetic diagnosis in 17 AS patients. No abnormal findings were observed when they were born. Developmental delay in movement, speech impairments and happy disposition were observed in 100% (17/17) AS patients. And the severe speech deficit was much easier and more obvious to observe than movement. About 80% (14/17) - 90% (15/17) AS patients presented frequent clinical characteristics, such as seizures and abnormal EEG. However, microcephaly could only be observed in 35% (6/17) AS patients. Regarding the associated findings of AS, 41% (7/17) - 77% (13/17) AS patients could be observed with flat occiput/occipital groove, prognathia, wide mouth, wide-spaced teeth, frequent drooling, excessive mouth behaviors, hypopigmented skin, light hair compared to parents, flexed arm position during ambulation and sleep disorder etc. These features occurred at a higher frequency in those patients of > 2 years old group than that of < 2 years old group.</p><p><b>CONCLUSION</b>The testing strategies of MS-PCR and STR linkage analysis combined with chromosome karyotype analysis were appropriate to the molecular genetic diagnosis of AS. In our analysis of clinical features, there was a lower rate of small head circumference (HC) in 35% patients compared with 80% patients in Caucasian with microcephaly, which might be attributable to the phenotypic heterogeneity in different races. And the birth history, movement and speech development and main clinical features of the Chinese AS patients were consistent with those of other studies. Clinical analysis in patients of different age groups showed that findings associated with AS would be more easily observed with the age increasing. Genetic diagnosis should be performed in clinically suspected AS patients.</p>

MR discrimination of early atypical tuberculous spondylitis from pyogenic spondylitis / 中国医学影像技术

Objective To detect the MRI manifestations and discrimination of tuberculous spondylitis and pyogenic spondylitis with atypical features in early stage. Methods Six patients with pathologically proved tuberculous spondylitis and 7 patients of pyogenic spondylitis with atypical clinical features and were included. MRI features of the vertebral bodies, intervertebral discs, paraspinal soft tissues and their enhancement patterns were analyzed. Chi-Square test was used to compare the MRI features of two diseases. Results Patients with pyogenic spondylitis had a significantly higher incidence of disk space narrowing (8 intervertebral bodies), abnormal signal in superior/inferior of vertebral body (12 intervertebral bodies) and endplate with high signal (13 intervertebral bodies), which were not seen in the patients with tuberculosis spondylitis (P<0.05).Patients with tuberculous spondylitis had a significantly higher incidence of local abnormal signal in anterior of vertebral body (4 intervertebral bodies) and paraspinal abscess spanning vertebral body (5 intervertebral bodies), while none of them was found in patients with pyogenic spondylitis (P<0.05). Conclusion MRI is accurate for the differentiation of tuberculous spondylitis and pyogenic spondylitis with atypical feature in early stage.

A study on genetic diagnosis for Angelman syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the genetic diagnosis of Angelman syndrome(AS), and provide information for clinic diagnosis and counseling to AS families.</p><p><b>METHODS</b>Methylation specific-PCR (MS-PCR) was used for primary diagnosis of 16 clinically suspected AS cases, and linkage analysis by short tandem repeat (STR) was applied to detect the molecular genetic defect in the nuclear families.</p><p><b>RESULTS</b>In this study, 10 AS patients were identified by MS-PCR, and 9 of them with maternal deletion in chromosome 15q11-q13, 1 with imprinting defect in chromosome 15q11-q13 were confirmed by STR linkage analysis.</p><p><b>CONCLUSION</b>Most of the AS patients could be confirmed by MS-PCR. And STR linkage analysis can detect the molecular defect of AS. It is very important for disease diagnosis, genetic counseling and prenatal diagnosis to perform the related genetic diagnosis.</p>

Mutation analysis and one novel mutation detection of 6-pyruvoyl tetrahydropterin synthase gene in children with tetrahydrobiopterin deficiency / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the distribution character of the mutations of 6-pyruvoyl tetrahydropterin synthase (PTPS) gene and to provide effective basis for gene diagnosis of tetrahydrobiopterin deficiency (BH4D) in children with hyperphenylalaninemia.</p><p><b>METHODS</b>Direct sequencing was performed for screening the PTPS gene mutations in 5 patients with clinically suspected BH4D and their parents. The nature of the novel mutations were deduced by the sequences alignment and the structural analysis of mutant protein. Artificial construct restriction site was used to detect the novel mutation in the control samples. The dates of urinary pterin analysis and BH4 loading test were retrospectively analyzed after gene analysis.</p><p><b>RESULTS</b>Four PTPS gene mutations (N52S, P87S, D96N, and L127F) were detected in our study. The genotypes of four PTPS deficiency patients were identified as N52S/L127F, P87S/D96N, N52S/D96N, and D96N/ -. As a novel mutation that has not been reported previously, the mutation L127F was not detected in 50 normal controls. This novel mutation L127F was inherited from the patient's mother, and this mutant site was highly conserved by sequences alignment and the protein structural analysis. Four of the five cases with hyperphenylalaninemia and suspicious BH4D, whose urinary biopterin percentage was lower than 2% , were diagnosed as PTPS deficiency during 5-20 months old. The remaining one case was excluded from BH4D.</p><p><b>CONCLUSIONS</b>The mutant characterization of PTPS gene was coincident with other early studies in Chinese. The novel mutation L127F was considered as a pathogenetic mutation and associated with severe clinical phenotype.</p>