AIM： To describe a Chinese family affected by a severe form of Axenfeld-Rieger syndrome （ARS） and characterize the molecular defect in PITX2 in the family. METHODS： Patients presented with typical ARS from a Chin...AIM： To describe a Chinese family affected by a severe form of Axenfeld-Rieger syndrome （ARS） and characterize the molecular defect in PITX2 in the family. METHODS： Patients presented with typical ARS from a Chinese family were investigated. We performed genome- wide linkage scan and exome sequencing to identify the pathogenic mutations, Candidate mutations were verified for co-segregation in the whole pedigree using Sanger sequencing, Real-time polymerase chain reaction （RT- PCR） and Western blotting were performed to verify the expression of the pathogenic gene. RESULTS： Genome-wide linkage and exome sequencing analyses showed PITX2 as the disease candidate gene. A〉G substitution at position -11 of 3＇ss of exon 5 （IVS5- 11A〉G） that co-segregated with the disease phenotype was discovered in the family. The PITX2 messenger ribonucleic acid and protein levels were about 50% lower in patients with ARS than in unaffected family members in the family, CONCLUSION： Our findings implicate the first intronic mutation of the PITX2 gene in the pathogenesis of a severe form of ARS in a Chinese family. This study highlights the importance of a systematic search for intronic mutation in ARS cases for which no mutations in the exons of PITX2 have been found.展开更多
Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but imp...Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but improvements are still needed. However, the ABCA12 null mice failed to respond to retinoid treatment, which impedes the development of novel cure strategies for HI. Here we generated an ethylnitrosourea mutagenic HI pig model (named Z9), which carries a novel deep intronic mutation IVS49-727 A>G in the ABCA12 gene, resulting in abnormal mRNA splicing and truncated protein production. Z9 pigs exhibit significant clinical symptom as human patients with HI. Most importantly, systemic retinoid treatment significantly prolonged the life span of the mutant pigs via improving epidermal maturation, decreasing epidermal apoptosis, and triggering the expression of ABCA6. Taken together, this pig model perfectly resembles the clinical symptom and molecular pathology of patients with HI and will be useful for understanding mechanistic insight and developing therapeutic strategies.展开更多
Lipoprotein lipase (LPL) gene is a multifunctional protein, playing a major role in the hydrolysis of triglycerides in chylomicrons and very low density lipoproteins (VLDL). According to cDNA of Landrace breed (GenBan...Lipoprotein lipase (LPL) gene is a multifunctional protein, playing a major role in the hydrolysis of triglycerides in chylomicrons and very low density lipoproteins (VLDL). According to cDNA of Landrace breed (GenBank accession: X62984), a pair of primers was designed for amplification of intron 3. Sequencing analysis indicated that a G1200A exits in Large White breed by cloning and sequencing. The mutation can be detected by PCR-EcoT22I-RFLP. Polymorphism analysis in a resource family showed that significant difference exits in carcass traits. Pigs with AA genotype had more 6th-7th rib fat thickness (13%,P< 0.05), thorax-waist-fat thickness (11.7%,P=0.065 3), buttock fat thickness (13.1%,P=0.073 0) and average fat thickness (10.4%,P=0.050 3) than pigs with BB genotype. LPL gene showed mainly in the pattern of additive effect and all the dominant effect were not significant. The value of additive effect of 6th-7th rib fat thickness, buttock fat thickness and average fat thickness was -0.17±0.07(P< 0.05),-0.12±0.06( P< 0.05),-0.12±0.06(P< 0.05),respectively. The allelic frequency is significantly different between indigenous Chinese breeds and European breeds.展开更多
The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis ...The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis of Parkinson＇s disease,30 Han Chinese patients with early-onset Parkinson＇s disease and 38 normal controls were studied to determine the sequence changes of 1,4,6 and 7 exon sections.In the 30 patients with Parkinson＇s disease,a heterozygous intron mutation（nt 119,G→G/A）in exon 1 was detected in one case;a homozygous intron mutation（nt 526500,T→C）between intron 3 and exon 4 in fourteen cases was found;a heterozygous intron mutation（nt 526607,G→G/A）between intron 3 and exon 4 was observed in eight cases;an exon 6missense mutation（nt 754317,C→C/T;codon 193,CGG→CGG/TGG;aa 193,Arg→Arg/Trp）in three cases was seen;and an exon 7 missense mutation（nt 941943,C→A/C;codon 272,CTC→CTC/ATC;aa 272,Leu→Leu/lle）was found in one case.These changes were not found in the normal population.The results indicated that the PARK2 exons 6 and 7 mutations are possibly pathogenic mutations,along with the intron 3-exert 4 and exon 1 mutations.PARK2 gene mutations are possible factors leading to the onset of Parkinson＇s disease.展开更多
Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in ...Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in intron 1 and Parkinson’s disease （PD） .Methods We screened all 7exons and exon-intron boundary regions of DJ-1 by PCR and direct nucleotide sequencing in 90 Chinese patients with EOPD.We also compared the allele and genotype frequencies of the g.168<sub>1</sub>85del polymorphism展开更多
基金Supported by China Postdoctoral Science Foundation Funded Project(No.2017M612211)the National Natural Science Foundation of China(No.81300742+2 种基金No.81600721)the Shandong Province Medical and Health Technology Development Project(No.2016WS0265)the Science and Technology Plan of Qingdao(No.15-9-1-35-jch)
文摘AIM： To describe a Chinese family affected by a severe form of Axenfeld-Rieger syndrome （ARS） and characterize the molecular defect in PITX2 in the family. METHODS： Patients presented with typical ARS from a Chinese family were investigated. We performed genome- wide linkage scan and exome sequencing to identify the pathogenic mutations, Candidate mutations were verified for co-segregation in the whole pedigree using Sanger sequencing, Real-time polymerase chain reaction （RT- PCR） and Western blotting were performed to verify the expression of the pathogenic gene. RESULTS： Genome-wide linkage and exome sequencing analyses showed PITX2 as the disease candidate gene. A〉G substitution at position -11 of 3＇ss of exon 5 （IVS5- 11A〉G） that co-segregated with the disease phenotype was discovered in the family. The PITX2 messenger ribonucleic acid and protein levels were about 50% lower in patients with ARS than in unaffected family members in the family, CONCLUSION： Our findings implicate the first intronic mutation of the PITX2 gene in the pathogenesis of a severe form of ARS in a Chinese family. This study highlights the importance of a systematic search for intronic mutation in ARS cases for which no mutations in the exons of PITX2 have been found.
基金the Strategic Priority Research Programs of CAS(XDA16030300)the National Natural Science Foundation of China(81671274,31272440,and 31801031)+1 种基金the National Transgenic Project of China(2016ZX08009003-006-007)the Elite Youth Program of the Chinese Academy of Agricultural Sciences(ASTIP-IAS05).
文摘Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but improvements are still needed. However, the ABCA12 null mice failed to respond to retinoid treatment, which impedes the development of novel cure strategies for HI. Here we generated an ethylnitrosourea mutagenic HI pig model (named Z9), which carries a novel deep intronic mutation IVS49-727 A>G in the ABCA12 gene, resulting in abnormal mRNA splicing and truncated protein production. Z9 pigs exhibit significant clinical symptom as human patients with HI. Most importantly, systemic retinoid treatment significantly prolonged the life span of the mutant pigs via improving epidermal maturation, decreasing epidermal apoptosis, and triggering the expression of ABCA6. Taken together, this pig model perfectly resembles the clinical symptom and molecular pathology of patients with HI and will be useful for understanding mechanistic insight and developing therapeutic strategies.
文摘Lipoprotein lipase (LPL) gene is a multifunctional protein, playing a major role in the hydrolysis of triglycerides in chylomicrons and very low density lipoproteins (VLDL). According to cDNA of Landrace breed (GenBank accession: X62984), a pair of primers was designed for amplification of intron 3. Sequencing analysis indicated that a G1200A exits in Large White breed by cloning and sequencing. The mutation can be detected by PCR-EcoT22I-RFLP. Polymorphism analysis in a resource family showed that significant difference exits in carcass traits. Pigs with AA genotype had more 6th-7th rib fat thickness (13%,P< 0.05), thorax-waist-fat thickness (11.7%,P=0.065 3), buttock fat thickness (13.1%,P=0.073 0) and average fat thickness (10.4%,P=0.050 3) than pigs with BB genotype. LPL gene showed mainly in the pattern of additive effect and all the dominant effect were not significant. The value of additive effect of 6th-7th rib fat thickness, buttock fat thickness and average fat thickness was -0.17±0.07(P< 0.05),-0.12±0.06( P< 0.05),-0.12±0.06(P< 0.05),respectively. The allelic frequency is significantly different between indigenous Chinese breeds and European breeds.
文摘The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis of Parkinson＇s disease,30 Han Chinese patients with early-onset Parkinson＇s disease and 38 normal controls were studied to determine the sequence changes of 1,4,6 and 7 exon sections.In the 30 patients with Parkinson＇s disease,a heterozygous intron mutation（nt 119,G→G/A）in exon 1 was detected in one case;a homozygous intron mutation（nt 526500,T→C）between intron 3 and exon 4 in fourteen cases was found;a heterozygous intron mutation（nt 526607,G→G/A）between intron 3 and exon 4 was observed in eight cases;an exon 6missense mutation（nt 754317,C→C/T;codon 193,CGG→CGG/TGG;aa 193,Arg→Arg/Trp）in three cases was seen;and an exon 7 missense mutation（nt 941943,C→A/C;codon 272,CTC→CTC/ATC;aa 272,Leu→Leu/lle）was found in one case.These changes were not found in the normal population.The results indicated that the PARK2 exons 6 and 7 mutations are possibly pathogenic mutations,along with the intron 3-exert 4 and exon 1 mutations.PARK2 gene mutations are possible factors leading to the onset of Parkinson＇s disease.
文摘Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in intron 1 and Parkinson’s disease （PD） .Methods We screened all 7exons and exon-intron boundary regions of DJ-1 by PCR and direct nucleotide sequencing in 90 Chinese patients with EOPD.We also compared the allele and genotype frequencies of the g.168<sub>1</sub>85del polymorphism
