发作性运动诱发性运动障碍一家系致病基因定位与突变分析

Objective To study the clinical characteristics and genetic cause of a Chinese family affected with paroxysmal kinesigenic dystonia(PKD).Methods The detailed clinical data and the blood samples of the affected patients with PKD and their relatives were collected.After genomic DNA was extracted from blood leukocytes,target linkage analysis Was performed using multiplex PCR by microsatellite marker's located in the reported critical region on chromosome 16.All exons and flanking regions of SCNN1G and ITGAL genes were amplified by PCR-sequence.Results In this three-generation 12 member family,5 individuals have been diagnosed as PKD.Target linkage analysis suggested the disease gene linked to chromosome 16.between D16S3396 and D16S3057 with two-point LOD score of 1.47 at recombination fraction(θ)=0.0.All affected individuals shared a common haplotype which co-segregated with the phenotype.Except for 8 reported SNPs,no pathologic sequence variants were found in candidate genes SCNN1G and ITGAL.Conclusions The studied family is genetically linked to the reported critical locus of PKD on chromosome 16.SCNN1G and ITGAL were ruled out as the causative genes for the studied pedigree.Further genetic analysis in this family may reveal new genetic cause responsible for PKD.     

遗传性痉挛性截瘫一家系的临床特点与遗传学分析

Objective To study the clinical and genetic features of a family with hereditary spastic paraplegia(HSP).Methods The patients were from a large Linyi family.Five members were clinically diagnosed with HSP according to Harding's criteria Blood samples were collected from family members.Genomic DNA was extracted from total blood samples using a standard phenol-chloroform extraction.The genetic linkage analysis was performed using microsatellite markers.Two-point linkage analysis was performed using the LINKAGE program.Five members underwent detailed neurological examinations and 4 members underwent electrophysiological analysis,cervical and thoracic MRI and serum enzymes.Results Linkage analysis mapped the AD-HSP locus to chromosome 2p12(SPG31)in this family.Positive LOD scores were obtained for SPG31 markers on chromosome 2 with a maximum multipoint LOD score of Z=1.8.Analysis of the REEP1 gene revealed a heterozygous G-to-A mutation at nucleotide position c417+1 donor site in exon 5.resulting in splice-site mutation.The symptoms of the patients manifested as stiffness,instability or weakness of the legs.MRI of the thoracic revealed atrophies of the spinal cord in the proband's son.Conclusions SPC31 patients have the clinical features of the typical HSP characteristics.REEP1 gene is the pathogenic gene.with REEP1 c417+1G＞A heterozygous mutation.     

Genetic relationship between serum pregnancy-associated plasma protein-A gene polymorphism and ischemic cerebrovascular disease in a Northern Han Chinese population

The present study recruited 193 patients with ischemic cerebrovascular disease from Inpatient and Outpatient Departments at the Affiliated Hospital of Qingdao University Medical College, China from August 2008 to May 2010, as well as 120 healthy volunteers from the Medical Examination Center at the Affiliated Hospital of Qingdao University Medical College, China, who served as controls for this study. Patients and control subjects were from the Han population in northern China. Enzyme- linked immunosorbent assay analysis revealed increased levels of serum pregnancy-associated plasma protein-A (PAPP-A) in ischemic cerebrovascular disease patients compared with healthy controls. In addition, the patients exhibited greater frequency of genotype CC and C alleles in a missense A/C (Tyr/Ser) polymorphism (dbSNP: rs7020782) of exon 14 in the PAPP-A gene. Multiple-factor logistic regression analysis on correction of age, gender, history of smoking, hypertension, diabetes mellitus, hypercholesteremia, and ischemic stroke family history showed that the risk for ischemic cerebrovascular disease in the population without the A allele at the A/C genetic locus in exon 14 of the PAPP-A was 2-folds greater than the population expressing the A allele. These experimental findings suggested that ischemic cerebrovascular disease correlated with the C allele in exon 14 of PAPP-A. In addition, the A allele is likely a protective gene; individuals carrying the A allele were less prone to ischemic cerebrovascular disease compared with individuals without the A allele.     

Risk factors for Parkinson disease and the path analysis: One-to-one paired design

BACKGROUND: Parkinson disease (PD) results from the reduce of neurotransmitter dopamine that transmits intracellular information in brain caused by some reasons, then leads to the dynamic disequilibrium with another neurotransmitter of acetylcholine which is relatively hyperactive. The main causes for PD are still unclear. OBJECTIVE: To screen out the risk factors of PD by means of univariate analysis and multivariate Logistic regression analysis, and investigate the manner of actions between various factors and PD, so as to provide clues for the etiological study of PD. DESIGN: A paired design, Logistic regression analysis, path analysis. SETTING: Department of Scientific Research, Shandong Institute of Physical Education. PARTICIPANTS: Totally 157 PD patients were selected from the Department of Neurology, Qilu Hospital of Shandong University from November 2001 to October 2002. Inclusive criteria: PD was diagnosed according to the standard set by the Fourth National Seminar on Extrapyramidal Disease, Parkinsonian syndromes caused by stroke, carbon monoxide poisoning, encephalitis, drugs, etc. were excluded. Another 157 patients treated in the same department at the same period were selected as the control group, they were the same in sex as those in the patient group, within 3 years older or younger than those in the patient group, and without PD or other extrapyramidal diseases. METHODS: ① The general conditions were investigated in all the subjects, including general conditions, social behavioral factor, environmental factor, genetic factor, life events, and previous disease; There were 12 main variables, including educational level, family history, mental labour, contact to insecticides, living place before school-age, smoking index, drinking index, tea-drinking index, history of brain trauma, history of cardiovascular disease, history of diabetes mellitus, and history of depression. ② SAS6.12 software and SPSS 10.0 software were used in the conditional Logistic regression analysis and path analysis respectively. MAIN OUTCOME MEASURES: The results of 12-variable univariate and multivariate analyses; Correlation between main variables and PD; Effects of the factors. RESULTS: All the subjects were involved in the analysis of results. ① The results of Logistic regression analysis showed that family history, mental labour, insecticides, drinking index and history of depression all had significant positive correlations with PD (univariate analysis: OR=1.405–5.429, P < 0.05–0.01; multivariate analysis: OR=2.029–6.754, P < 0.05–0.01), whereas smoking had significant negative correlations with PD [univariate analysis: odd ratio (OR)=0.765, P < 0.05; multivariate analysis: OR =0.489, P < 0.01]. ② The path analysis showed that family history, mental labour, insecticides, smoking, drinking and history of depression had direct effects on PD occurrence [(path coefficient=–0.218 to 0.204, P < 0.05–0.01)]; Insecticides could cause PD indirectly on the basis of family history (genetic susceptibility) (path coefficient=0.946, P < 0.01); Insecticides could also cause PD by drinking (path coefficient=0.165, P < 0.01); Drinking could cause PD indirectly on the basis of family history (path coefficient=0.043, P < 0.01). CONCLUSION: The main risk factors of PD are family history, history of depression, drinking, mental labour and insecticides, whereas the protective factor is smoking. PD attack has genetic susceptibility, insecticides and drinking can cause PD on the basis of PD family history. The risk of PD can be decreased by reducing the occasion for contacting the environmental risk factors.     

Risk factors for Parkinson disease and the path analysis： One-to-one paired design

BACKGROUND： Parkinson disease （PD） results from the reduce of neurotransmitter dopamine that transmits intracellular information in brain caused by some reasons, then leads to the dynamic disequilibrium with another neurotransmitter of acetylcholine which is relatively hyperactive. The main causes for PD are still unclear. OBJECTIVE： To screen out the risk factors of PD by means of univariate analysis and multivariate Logistic regression analysis, and investigate the manner of actions between various factors and PD, so as to provide clues for the etiological study of PD. DESIGN： A paired design, Logistic regression analysis, path analysis. SETTING： Department of Scientific Research, Shandong Institute of Physical Education. PARTICIPANTS： Totally 157 PD patients were selected from the Department of Neurology, Qilu Hospital of Shandong University from November 2001 to October 2002. Inclusive criteria： PD was diagnosed according to the standard set by the Fourth National Seminar on Extrapyramidal Disease, Parkinsonian syndromes caused by stroke, carbon monoxide poisoning, encephalitis, drugs, etc. were excluded. Another 157 patients treated in the same department at the same period were selected as the control group, they were the same in sex as those in the patient group, within 3 years older or younger than those in the patient group, and without PD or other extrapyramidal diseases. METHODS： （1） The general conditions were investigated in all the subjects, including general conditions, social behavioral factor, environmental factor, genetic factor, life events, and previous disease; There were 12 main variables, including educational level, family history, mental labour, contact to insecticides, living place before school-age, smoking index, drinking index, tea-drinking index, history of brain trauma, history of cardiovascular disease, history of diabetes mellitus, and history of depression. （2） SAS6.12 software and SPSS 10.0 software were used in the conditional Logistic regression analysis and path analysis respectively. MAIN OUTCOME MEASURES： The results of 12-variable univariate and multivariate analyses; Correlation between main variables and PD; Effects of the factors. RESULTS： All the subjects were involved in the analysis of results. （1） The results of Logistic regression analysis showed that family history, mental labour, insecticides, drinking index and history of depression all had significant positive correlations with PD （univariate analysis： OR=1.405- 5.429, P 〈 0.05- 0.01; multivariate analysis： OR=2.029- 6.754, P 〈 0.05- 0.01）, whereas smoking had significant negative correlations with PD [univariate analysis： odd ratio （OR）=0.765, P 〈 0.05; multivariate analysis： OR =0.489, P 〈 0.01]. （2） The path analysis showed that family history, mental labour, insecticides, smoking, drinking and history of depression had direct effects on PD occurrence [（path coefficient= - 0.218 to 0.204, P 〈 0.05 -0.01）]; Insecticides could cause PD indirectly on the basis of family history （genetic susceptibility） （path coefficient=0.946, P 〈 0.01）; Insecticides could also cause PD by drinking （path coefficient=0.165, P 〈 0.01） Drinking could cause PD indirectly on the basis of family history （path coefficient=0.043, P 〈 0.01 ）. CONCLUSION： The main risk factors of PD are family history, history of depression, drinking, mental labour and insecticides, whereas the protective factor is smoking. PD attack has genetic susceptibility, insecticides and drinking can cause PD on the basis of PD family history. The risk of PD can be decreased by reducing the occasion for contacting the environmental risk factors.     

中性脂肪沉积症合并肌病一家系

Objective To study the clinical,myopathological features in neutral lipid storage disease with myopathy (NLSDM) caused by a novel PNPLA2 mutation.Methods Two patients were siblings.The proband was a 40-year-old woman.She presented progressive limb weakness and muscle atrophy at 35 years old.Her 55-year-old brother presented deafness at 35 years old and limb weakness at 45 years old. He suffered from ventricular septal defect.Open biosies were performed on them and specimens were studied histologically enzymhistochemically.and ultrastructurally.All the exons of PNPLA2 gene were analyzed in the both patients and 3 healthy family individuals.Results Muscle biopsy in both patients revealed hypertrophy and atrophy of fibers with proliferation of connective tissue.There were numerous lipid droplets and plenty of rimmed vacuoles in the fibers.Electron microscopy revealed lipid droplets between sarcomeres as well as myelin figures.A single homozygous base substitution was detected at the beginning of intron 2(IVS2+1G＞A)of PNPLA2 in two patients.but not in the healthy family individuals. Conclusion The novel IVS2+1G＞A mutation of PNPLA2 causes NLSDM with prominent limb weakness.The disease may be associated with auditory nerve lesions and congenital heart disense.Rimmed vacuoles with lipid storage in the fibets might have indication for diagnosis of NLSDM.     

Cerebral vasculopathy in a Chinese family with neurofibromatosis type Ⅰ mutation

Neurofibromatosis type I（NF1） is a hereditary,autosomal dominant,neurocutaneous syndrome that is attributed to NF1 gene mutation.NF1 has been associated with scoliosis,macrocephaly,pseudoarthrosis,short stature,mental retardation,and malignancies.NF1-associated vasculopathy is an uncommon and easily-overlooked presentation.Examination of a Chinese family affected by NF1 combined with cerebral vessel stenosis and/ or abnormality suggested a possible relationship between NF1 and vessel stenosis.To determine which NF1 gene mutation is associated with vascular lesions,particularly cerebral vessel stenosis,we examined one rare family with combined cerebral vessel lesions or maldevelopment.Vascular lesions were detected using transcranial Doppler sonography and digital subtraction angiography in family members.Next,denaturing high-performance liquid chromatography and sequencing were used to screen for NF1 gene mutations.The results revealed a nonsense mutation,c.541C＆gt;T,in the NF1 gene.This mutation truncated the NF1 protein by 2659 aminoacid residues at the C-terminus and co-segregated with all of the patients,but was not present in unaffected individuals in the family.Exceptionally,three novel mutations were identified in unaffected family members,but these did not affect the product of the NF1 gene.Thus the nonsense mutation,c.541C＆gt;T,located in the NF1 gene could constitute one genetic factor for cerebral vessel lesions.     

PARK1 gene mutation of autosomal dominant Parkinson’s disease family

Studies have shown that PARK1 gene is associated with the autosomal dominant inheritance of Parkinson’s disease. PARK1 gene contains two mutation sites, namely Ala30Pro and Ala53Thr, which are located on exons 3 and 4, respectively. However, the genetic loci of the pathogenic genes remain unclear. In this study, blood samples were collected from 11 members of a family with high prevalence of Parkinson’s disease, including four affected cases, five suspected cases, and two non-affected cases. Point mutation screening of common mutation sites on PARK1 gene exon 4 was conducted using PCR, to determine the genetic loci of the causative gene for Parkinson’s disease. Gene identification and sequencing results showed that a T base deletion mutation was observed in the PARK1 gene exon 4 of all 11 collected samples. It was confirmed that the PARK1 gene exon 4 gene mutation is an important pathogenic mutation for Parkinson’s disease.     

Challenges in studying geographic atrophy(GA) age-related macular degeneration: the potential of a new mouse model with GA-like features

Loss of central vision critical to everyday activities such as reading,face-recognition and driving due to damage in the central retina (the macula) is the leading cause of irreversible blindness amongst adults in the developed world.This condition,termed age-related macular degeneration (AMD),is a complex,chronic degenerative disease driven by a combination of genetic and lifestyle risk factors.Early signs of retinal changes in people as young as 30–40 years have been reported,although these individuals appear to be asymptomatic.However,by the age of 65,the disease is present in ~3% of individuals,which increases dramatically to affect 1/3 of individuals by the eighth decade of life.Early to intermediate AMD is estimated to affect ~150 million individuals globally,with another 10 million individuals suffering from end-stage,sight-threatening forms.These terminal stages are broadly grouped into dry (geographic atrophy,GA) or wet (choroidal neovascular,CNV) AMD (Sarks et al.,1988;Bird et al.,2014),with similar frequencies reported in patients.Recent advances in identifying genetic risk factors,including our discoveries in this field,indicate an initial shared pathology before progressing to aforementioned late-stage phenotypes.Currently,GA patients have no effective treatment,which may in part be due to the lack of good in vivo models for GA studies.Here,we summarize our new findings that describe an altogether new mouse model with GA-like features which shows progressive outer retinal pathology (Ibbett et al.,2019) that can be used to gain novel insights into GA and potentially as a tool for drug development.     

Paroxysmal kinesigenic dyskinesia and myotonia congenita in the same family：coexistence of a PRRT2 mutation and two CLCN1 mutations

Paroxysmal kinesigenic dyskinesia（PKD） and myotonia congenita（MC） are independent disorders that share some clinical features. We aimed to investigate the sequences of PRRT2 and CLCN1 in a proband diagnosed with PKD and suspected MC. Clinical evaluation and auxiliary examinations were performed. Direct sequencing of the entire coding regions of the PRRT2 and CLCN1 genes was conducted. Haplotype analysis confirmed the relationships among the family members. The proband suffered choreoathetosis attacks triggered by sudden movements, and lower-limb weakness a n d s t i ff n e s s t h a t w o r s e n e d i n c o l d w e a t h e r. Carbamazepine monotherapy completely controlled his choreoathetosis and significantly relieved his limb weakness and stiffness. His father, when young, had similar limb stiffness, while his mother and brother were asymptomatic. Genetic analysis revealed that the proband and his father harbored a PRRT2 c.649 dup C mutation, and CLCN1 c.1723C〉T and c.2492A〉G mutations. His brother carried only the two CLCN1 mutations. None of these mutations were identified in his mother and 150 unrelated controls. This is the first report showing the coexistence ofPRRT2 and CLCN1 mutations. Our results also indicate that both the PRRT2 and CLCN1 genes need to be screened if we fail to identify PRRT2 mutations in PKD patients or CLCN1 mutations in MC patients.