中国Rett综合征患儿突变基因的亲源分析

Objective To identify the parental origin of methyl-CpG-binding protein 2 (MECP2)gene mutations in Chinese patients with Rett syndrome. Methods Single nucleotide polymorphisms (SNPs) in intron 3 of the MECP2 gene were analyzed by PCR and sequencing in 115 patients with Rett syndrome.Then sequencing of the SNP region was performed for the fathers of the patients who had at least one SNP,to determine which allele was from the father. Then allele-specific PCR was performed and the products were sequenced to see whether the allele from father or mother harbored the mutation. Results Seventy-six of the 115 patients had at least one SNP. Three hot SNPs were found in these patients. They were: IVS3+22C＞G, IVS3+266C＞T and IVS3+683C＞T. Among the 76 cases, 73 had a paternal origin of MECP2 mutations, and the other 3 had a maternal origin. There were multiple types of MECP2 mutation of the paternal origin, including 4 frame shift, 2 deletion and 67 point (56 C＞T, 6 C＞G, 2 A＞G, 2 G＞T and 1 A＞T) mutations. The mutation types of the 3 ptients with maternal origin included 2 frame shift and 1 point (C＞T) mutation. Conclusion In Chinese RTT patients, the MECP2 mutations are mostly of paternal origin.     

Mutation analysis of 11 Chinese patients with attenuated mucopolysaccharidosis type Ⅰ

Objective Mucopolysaccharidosis type Ⅰ (MPS Ⅰ ) is an autosomal recessive diseaseresulting from the deficiency in the lysosomal enzyme α-L-iduronidase (IDUA). The present study was conducted to identify IDUA gene mutations in attenuated (MPS Ⅰ H/S and MPS Ⅰ S) patients with MPS Ⅰin northern China. Methods Fourteen exons with adjacent intronic sequences of the IDUA gene in 11 MPS Ⅰ patients were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced directly and origin analysis was conducted. Results Seven mutations were detected in the 11 MPS Ⅰ patients, i.e., c. 236 C＞T(p. A79V), c. 266 G＞A(p. R89Q), c. 265 C＞T(p. R89W), c. 532G＞A(p.E178K), c. 589G＞A(p. G197S), c. 1037T＞G(p. L346R), and c. 1877 G＞A(p. W626X). All of them were known mutations. Six patients were homozygotes and 1 was heterozygote with nonsense mutation. In addition, 9 reported single nucleotide polymorphism (SNP) were detected, i. e., p. A8, p. A20, p. H33Q,p. R105Q, p. A314, p. A361T, p. T388, p. T410 and p. V454I. Conclusion The mutation spectrum of the IDUA gene in attenuated MPS Ⅰ Chinese patients may be different from that in patients from other countries.     

Mutation analysis of 11 Chinese patients with attenuated mucopolysaccharidosis type Ⅰ

Objective Mucopolysaccharidosis type Ⅰ (MPS Ⅰ ) is an autosomal recessive diseaseresulting from the deficiency in the lysosomal enzyme α-L-iduronidase (IDUA). The present study was conducted to identify IDUA gene mutations in attenuated (MPS Ⅰ H/S and MPS Ⅰ S) patients with MPS Ⅰin northern China. Methods Fourteen exons with adjacent intronic sequences of the IDUA gene in 11 MPS Ⅰ patients were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced directly and origin analysis was conducted. Results Seven mutations were detected in the 11 MPS Ⅰ patients, i.e., c. 236 C＞T(p. A79V), c. 266 G＞A(p. R89Q), c. 265 C＞T(p. R89W), c. 532G＞A(p.E178K), c. 589G＞A(p. G197S), c. 1037T＞G(p. L346R), and c. 1877 G＞A(p. W626X). All of them were known mutations. Six patients were homozygotes and 1 was heterozygote with nonsense mutation. In addition, 9 reported single nucleotide polymorphism (SNP) were detected, i. e., p. A8, p. A20, p. H33Q,p. R105Q, p. A314, p. A361T, p. T388, p. T410 and p. V454I. Conclusion The mutation spectrum of the IDUA gene in attenuated MPS Ⅰ Chinese patients may be different from that in patients from other countries.     

Mutation screening of the F Ⅷ gene in 10 hemophilia A families

Objective To identify the F Ⅷ gene mutations of patients and suspected female carriers in 10 Hemophilia A (HA) families, and to guide the prenatal diagnosis. Methods PCR, denaturing high performance liquid chromatogramphy(DHPLC) and DNA sequencing technologies were applied to screen the FⅧ gene of 8 HA patients and 12 suspected female carriers in the 10 families. Linkage analysis was performed by using St 14 (DXS 52), intron 13 (CA)n and EX18/Bcl Ⅰ of the FⅧ gene in the HA families.In prenatal diagnosis, we screened the same mutation found in the patients. PCR-restriction fragment length polymorphism was applied to detect the new missense mutations of F Ⅷ gene in 100 unrelated healthy individuals to exclude the possibility of polymorphism. Results (1) Five missense mutations, 3 frameshift mutations, 2 nonsense mutations and 2 single nucleotide polymorphism(SNP) were identified in 10 the HA families. Among them, c. 878A＞G, c. 1015A＞G, c. 6870G＞T, c. 1282delA, c. 3072_3073insT, c. 4880_4881insA and c. 5000G＞A were novel mutations or polymorphism. No missense mutations c. 878A＞G, c.1015A＞G and c. 6870G＞T, were found in the 100 healthy unrelated controls. (2) Nine suspected female carriers were confirmed at the gene level. (3) X risk chromosome could be determined in 4 HA families by genetic linkage analysis. (4) Among the four fetuses for prenatal diagnosis, 2 were normal, 1 was carrier and the remaining 1 was a patient. Conclusion Six novel mutations, i. e. , c. 878A＞G, c. 1015A＞G, c.6870G＞T, c. 1282delA, c. 3072_3073insT and c. 4880_4881insA, were identified in this study. PCR,DHPLC and DNA sequencing could be used to screen the gene mutations of HA patients, to carry out carrier detection and prenatal diagnosis of HA families efficiently, by combining with restriction endonuclease analysis and genetic linkage analysis.     

Mutation screening of the F Ⅷ gene in 10 hemophilia A families

Objective To identify the F Ⅷ gene mutations of patients and suspected female carriers in 10 Hemophilia A (HA) families, and to guide the prenatal diagnosis. Methods PCR, denaturing high performance liquid chromatogramphy(DHPLC) and DNA sequencing technologies were applied to screen the FⅧ gene of 8 HA patients and 12 suspected female carriers in the 10 families. Linkage analysis was performed by using St 14 (DXS 52), intron 13 (CA)n and EX18/Bcl Ⅰ of the FⅧ gene in the HA families.In prenatal diagnosis, we screened the same mutation found in the patients. PCR-restriction fragment length polymorphism was applied to detect the new missense mutations of F Ⅷ gene in 100 unrelated healthy individuals to exclude the possibility of polymorphism. Results (1) Five missense mutations, 3 frameshift mutations, 2 nonsense mutations and 2 single nucleotide polymorphism(SNP) were identified in 10 the HA families. Among them, c. 878A＞G, c. 1015A＞G, c. 6870G＞T, c. 1282delA, c. 3072_3073insT, c. 4880_4881insA and c. 5000G＞A were novel mutations or polymorphism. No missense mutations c. 878A＞G, c.1015A＞G and c. 6870G＞T, were found in the 100 healthy unrelated controls. (2) Nine suspected female carriers were confirmed at the gene level. (3) X risk chromosome could be determined in 4 HA families by genetic linkage analysis. (4) Among the four fetuses for prenatal diagnosis, 2 were normal, 1 was carrier and the remaining 1 was a patient. Conclusion Six novel mutations, i. e. , c. 878A＞G, c. 1015A＞G, c.6870G＞T, c. 1282delA, c. 3072_3073insT and c. 4880_4881insA, were identified in this study. PCR,DHPLC and DNA sequencing could be used to screen the gene mutations of HA patients, to carry out carrier detection and prenatal diagnosis of HA families efficiently, by combining with restriction endonuclease analysis and genetic linkage analysis.     

Genetic analysis of two children patients affected with CHARGE syndromeCSCD

Objective To analyze two Chinese pediatric patients with multiple malformations and growth and development delay. Methods Both patients were subjected to targeted gene sequencing, and the results were analyzed with Ingenuity® Variant Analysis™ software. Suspected pathogenic variations were verified by Sanger sequencing. Results High-throughput sequencing showed that both patients have carried heterozygous variants of theCHD7 gene. Patient 1 carried a nonsense mutation in exon 36 (c. 79570T, p. Arg2653 ∗), while patient 2 carried a nonsense mutation of exon 2 (c. 7180T, p. Gln240 ∗ ). Sanger sequencing confirmed the above mutations in both patients, while their parents were of wild-type for the corresponding sites, indicating that the two mutations have happened de novo. Conclusion Two patients were diagnosed with CHARGE syndrome by high-throughput sequencing. © 2018 West China University of Medical Sciences. All rights reserved.     

河南地区苯丙酮尿症患者苯丙氨酸羟化酶基因突变研究

Objective To study the characteristics of the phenylalanine hydroxylase gene (PAH)mutations in patients with phenylketonuria (PKU) in Henan province, in order to provide basic information for genetic counseling and prenatal diagnosis. Methods Mutations of the PAH gene were detected in exons 1-13 with flanking introns of PAH gene by PCR and DNA sequencing in 47 families with PKU. Results A total of 25 different mutations were detected in 83 out of 94 PAH alleles (88. 3%). Among them,E79fX13, H271R and D415Y have not been reported previously. It was the first time that IVS10-14C＞Gmutation was reported in Chinese PKU population. The mutations p. R243Q, EX6-96A＞G, p. Y356X,IVS4-1G＞A, p. R111X, p. V399V and p. R413P, were the prevalent mutations with relative frequencies of 20. 5 %, 12.0%, 9.6%, 9. 6%, 8. 4%, 8. 4% and 7.2% respectively. Conclusion The mutations of the PAH gene in patients with classical phenylketonuria in Henan province were similar to that in other areas of China. Prenatal gene diagnosis for PKU by PAH gene sequencing is efficient for most PKU families.     

河南地区苯丙酮尿症患者苯丙氨酸羟化酶基因突变研究

Objective To study the characteristics of the phenylalanine hydroxylase gene (PAH)mutations in patients with phenylketonuria (PKU) in Henan province, in order to provide basic information for genetic counseling and prenatal diagnosis. Methods Mutations of the PAH gene were detected in exons 1-13 with flanking introns of PAH gene by PCR and DNA sequencing in 47 families with PKU. Results A total of 25 different mutations were detected in 83 out of 94 PAH alleles (88. 3%). Among them,E79fX13, H271R and D415Y have not been reported previously. It was the first time that IVS10-14C＞Gmutation was reported in Chinese PKU population. The mutations p. R243Q, EX6-96A＞G, p. Y356X,IVS4-1G＞A, p. R111X, p. V399V and p. R413P, were the prevalent mutations with relative frequencies of 20. 5 %, 12.0%, 9.6%, 9. 6%, 8. 4%, 8. 4% and 7.2% respectively. Conclusion The mutations of the PAH gene in patients with classical phenylketonuria in Henan province were similar to that in other areas of China. Prenatal gene diagnosis for PKU by PAH gene sequencing is efficient for most PKU families.     

Citrin缺陷导致的新生儿肝内胆汁淤积症临床和SLC25A13基因突变的研究

Objectives To investigate the clinical and laboratory features of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and to characterize the molecular basis and prognosis of this disease. Methods Twenty-six patients with NICCD were collected because of idiopathic intrahepatic cholestasis and jaundice. The diagnosis was made by routine laboratory data collection, tandem mass spectrometry (MS-MS) and gas chromatography mass spectrometry (GC-MS) analyses. SLC25A13 gene mutation was analyzed by using polymerase chain reaction (PCR), direct DNA sequencing and restriction fragment length polymorphism analyses. The patients were followed up for nearly 2 years. Results The NICCD patients showed low birth weight and the average onset of jaundice was 29 days. Laboratory data showed liver dysfunction, hyperbilirubinemia, hypoproteinemia, high levels of α-fetoprotein, prolonged prothrombin time, hypoglycemia and hyperammonemia. MS-MS analysis of the blood samples revealed specific elevation of citrulline, methionine, threonine, tyrosine and elevation of free carnitine, short-chain and long-chain acylcarnitines. GC-MS analysis of the urine samples showed elevated 4-hydroxyl phenyllactic acid and 4-hydroxyi phenylpyruvic acid. Twelve different mutations were identified, including 4 novel mutations, i. e. , G386V, R467X, K453R and 1192-1193delT. Forty-four mutated alleles were identified in the 52 alleles (84.6%). Among them, 851del4, 1638ins23 and IVS6+5G＞A mutations were the most frequent mutations, accounting for 40.9%, 20.5% and 11.4% of the total alleles examined respectively.Five of the 26 patients have not been recovered, including 4died and 1 accepted liver transplantation. No obvious relationship was found between the genotype and phenotype in NICCD. Conclusion The 851del4,1638ins23 and IVS6+5G＞A mutations are the hot-spot mutations in Chinese NICCD patients. Some NICCD patients have poor prognosis.     

Citrin缺陷导致的新生儿肝内胆汁淤积症临床和SLC25A13基因突变的研究

Objectives To investigate the clinical and laboratory features of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and to characterize the molecular basis and prognosis of this disease. Methods Twenty-six patients with NICCD were collected because of idiopathic intrahepatic cholestasis and jaundice. The diagnosis was made by routine laboratory data collection, tandem mass spectrometry (MS-MS) and gas chromatography mass spectrometry (GC-MS) analyses. SLC25A13 gene mutation was analyzed by using polymerase chain reaction (PCR), direct DNA sequencing and restriction fragment length polymorphism analyses. The patients were followed up for nearly 2 years. Results The NICCD patients showed low birth weight and the average onset of jaundice was 29 days. Laboratory data showed liver dysfunction, hyperbilirubinemia, hypoproteinemia, high levels of α-fetoprotein, prolonged prothrombin time, hypoglycemia and hyperammonemia. MS-MS analysis of the blood samples revealed specific elevation of citrulline, methionine, threonine, tyrosine and elevation of free carnitine, short-chain and long-chain acylcarnitines. GC-MS analysis of the urine samples showed elevated 4-hydroxyl phenyllactic acid and 4-hydroxyi phenylpyruvic acid. Twelve different mutations were identified, including 4 novel mutations, i. e. , G386V, R467X, K453R and 1192-1193delT. Forty-four mutated alleles were identified in the 52 alleles (84.6%). Among them, 851del4, 1638ins23 and IVS6+5G＞A mutations were the most frequent mutations, accounting for 40.9%, 20.5% and 11.4% of the total alleles examined respectively.Five of the 26 patients have not been recovered, including 4died and 1 accepted liver transplantation. No obvious relationship was found between the genotype and phenotype in NICCD. Conclusion The 851del4,1638ins23 and IVS6+5G＞A mutations are the hot-spot mutations in Chinese NICCD patients. Some NICCD patients have poor prognosis.     

Modes of Inheritance of Errors of Refraction

Eighteen families in which both parents had refractions within the range of +4·0 D to −4·0 D and axial lengths seen in emmetropia (22·3-26·0 mm) showed coefficients of correlation of the order 0·5 indicative of polygenic inheritance. Such coefficients were seen for axial length (0·407) and for the cornea (0·487), but not for the lens (which is known to be yoked to the axial length). No such coefficients were seen in 19 families in which one of the parents had axial length outside the emmetropic range (nine families with long axes and 10 with short axes).

The pattern of polygenic inheritance for emmetropia (completely correlated optical components) and errors of refraction up to 4·0 D (inadequately correlated components: correlation ametropia) follows that seen in stature and other measurable characters. In contrast the high refractive errors with their abnormal axial lengths (component ametropia) are—like the extremes in stature—pathological anomalies with monofactorial inheritance.

     

Recovery of responsiveness of cells of lateral geniculate nucleus of rat

1. Recovery of responsiveness of single cells in lateral geniculate nucleus of rat has been determined in both P and I cells. There are three types of recovery curve among P cells; (a) early recovery, (b) early partial recovery followed by depression and then complete recovery, (c) prolonged depression followed by cyclic recovery. Type (c) is by far the commonest recovery curve. In contrast to the spike in a P cell, the synaptic potential recovers to its full amplitude in about 20 msec. All I cells exhibit similar rapid recovery curves after a prolonged depression.2. Conditioning stimuli applied to visual cortex also produce a prolonged depression in most P cells but I cells can be re-excited at short intervals from cortex. Decortication does not prevent the prolonged depression of the multineuronal response produced by optic nerve stimulation.3. A neuronal model is proposed to explain these observations. It is supposed that I cells (interneurones) are innervated by axon collaterals of the P cells (principal cells, projecting to visual cortex) and that the I cells exert an inhibitory influence on the P cells.     

Level of functioning of siblings and parents of probands of varying degrees of retardation

A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.     

局部注射辛伐他汀对骨质疏松大鼠股骨髁骨小梁的改建效应

背景：局部注射具有成骨作用的辛伐他汀,可显著增加骨质疏松大鼠股骨颈及股骨髁部的骨密度及力学强度,分析局部注射辛伐他汀对股骨髁骨小梁的影响。 目的：进一步研究骨质疏松大鼠股骨内局部注射辛伐他汀对股骨髁骨小梁的影响。为将辛伐他汀应用于临床骨质疏松局部治疗提供实验基础。 方法：18只雌性SD大鼠双侧卵巢切除后3个月,制备大鼠骨质疏松模型。实验大鼠随机数字表法均分为3组,分别在实验大鼠的右侧股骨髓腔内单次注射辛伐他汀溶液5 mg、10 mg,对照组单纯注射空白载体。分别在注射后1个月处死大鼠并取材。Micro-CT扫描并定量分析骨组织形态变化。 结果与结论：给药后1个月,Micro-CT扫描结果显示,辛伐他汀治疗组的骨微结构参数如骨皮质厚度、骨小梁密度及连接率明显优于对照组。说明疏松骨骼单次注射小剂量辛伐他汀可显著促进股骨髁部骨小梁改建,改善骨骼微结构,可为强化局部、防治骨质疏松骨折的新选择进一步提供实验基础。