Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

ObjectiveTo evaluate the efficacy and safety profiles of enteeavir (ETV) in patients with advanced schistosomiasis and hepatitis B virus (HBV) co-infection.Methods Totally sixty patients with advanced schistosomiasis and HBV co-infection were enrolled in this study.The patients were divided into ETV treatment group (n=30) and rhubarb treatment group who refused to receive antiviral treatment (n=30).The patients were treated with ETV or rhubarb thelepus ball on the basis of routine supportive therapy for 52 weeks.The hepatic fibrosis markers (e.g.hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin),alanine transaminase (ALT),HBV DNA,Child-Pugh score between two groups were compared.Intention to treat (ITT) population was used for analysis.The measurement data and the enumeration data were analyzed by t test and x2 test,respectively.ResultsAfter 52-week treatment,the hepatic fibrosis markers (hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin) were significantly improved in ETV treatment group compared to the rhubarb treatment group (t =3.952,3.765,3.857,3.122 and 3.735,respectively; all P＜0.05),and the fibrosis of liver tissue in ETV treatment group was significantly improved compared with rhubarb treatment group (x2 =11.207,P＜0.05).The ALT level,HBV DNA,Child-Pugh score after 52-weeks treatment in ETV treatment group were statistically reduced compared with rhubarb treatment group (t =3.287,4.382 and 3.872,respectively; all P＜0.05),meanwhile,the ALT normalization rate and HBV DNA undetectable rate were significantly increased in ETV treatment group (x2 =17.376 and 39.095,respectively; both P＜0.05).In addition,no obvious adverse reaction was observed during ETV treatment.Conclusion Entecavir is safe and effective in patients with advanced schistosomiasis and HBV co-infection.

Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene.Methods SMN1 gene was amplified specifically with double allele-specific PCR(AS-PCR).Meanwhile,one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes.Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately.Conclusion Compared to PCR-RFLP and DHPLC used in the past,this approach can diagnose homozygous deletion of SMA much more accurate,easier and more convenient without completed following analyses.

Objective:To learn about the iodine nutrition level and its spatial distribution status in key populations in Hubei Province, so as to provide a basis for adjustment of iodine supplementation policy and the realization of scientific and accurate iodine supplementation.Methods:In 2020, a sampling was carried out in Hubei Province according to the "National Iodine Deficiency Disorders Monitoring Plan (2016 Edition)" to monitor the concentration of salt iodine and urinary iodine of key populations (children ages 8 - 10 years old and pregnant women). The spatial distribution of iodine nutrition levels was analyzed by spatial epidemiology.Results:The median salt iodine of 17 263 children's family salt samples was 25.0 mg/kg, and the median urinary iodine (MUI) was 217.0 μg/L. There was significant spatial aggregation in the distribution of urinary iodine level in children at the county level ( Moran's Index = 0.36, P < 0.001). The significant hot spot areas with high urinary iodine level among children were located in Shiyan City and Xiangyang City, while the significant cold spot areas with low urinary iodine level were mainly concentrated in Yichang City. The median salt iodine of 8 618 pregnant women's family salt samples was 25.1 mg/kg, the MUI was 176.3 μg/L. The urinary iodine level among pregnant women at the county level was spatially clustered ( Moran's Index = 0.22, P = 0.003) . The significant hot spot areas with high urinary iodine level among pregnant women were mainly in Enshi Tujia and Miao Autonomous Prefecture, the significant cold spot areas were mainly concentrated in Yichang City. Conclusions:In 2020, the iodine nutrition of children in Hubei Province is at a super appropriate level (200 - 299 μg/L), and the iodine nutrition status of pregnant women is more sensitive, which is close to the lower limit of the appropriate level (150 μg/L). The urinary iodine level of children and pregnant women has significant spatial aggregation at the county level. Targeted intervention will be needed in counties (dictricts) where the urinary iodine level is lower or higher than the normal range, to achieve accurate and scientific iodine supplementation.

OBJECTIVETo characterize the mutation spectrum of phenylalanine hydroxylase (PAH) gene and perform prenatal diagnosis for families with classical phenylketonuria.

METHODSBy stratified sequencing, mutations were detected in the exons and flaking introns of PAH gene of 44 families with classical phenylketonuria. 47 fetuses were diagnosed by combined sequencing with linkage analysis of three common short tandem repeats (STR) (PAH-STR, PAH-26 and PAH-32) in the PAH gene.

RESULTSThirty-one types of mutations were identified. A total of 84 mutations were identified in 88 alleles (95.45%), in which the most common mutation have been R243Q (21.59%), EX6-96A>G (6.82%), IVS4-1G>A (5.86%) and IVS7+2T>A (5.86%). Most mutations were found in exons 3, 5, 6, 7, 11 and 12. The polymorphism information content (PIC) of these three STR markers was 0.71 (PAH-STR), 0.48 (PAH-26) and 0.40 (PAH-32), respectively. Prenatal diagnosis was performed successfully with the combined method in 47 fetuses of 44 classical phenylketonuria families. Among them, 11 (23.4%) were diagnosed as affected, 24 (51.1%) as carriers, and 12 (25.5%) as unaffected.

CONCLUSIONPrenatal diagnosis can be achieved efficiently and accurately by stratified sequencing of PAH gene and linkage analysis of STR for classical phenylketonuria families.

Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Fetal Diseases ; diagnosis ; enzymology ; genetics ; Genetic Testing ; Humans ; Infant ; Infant, Newborn ; Male ; Microsatellite Repeats ; Middle Aged ; Pedigree ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; diagnosis ; enzymology ; genetics ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; Young Adult

The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.