OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with multiple pterygium syndrome (MPS). METHODS: A child with MPS who was treated at the Orthopedics Department of Guangzhou Women and Children's Medical Center Affiliated to Guangzhou Medical University on August 19, 2020 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her parents were also collected. Whole exome sequencing (WES) was carried out for the child. Candidate variant was validated by Sanger sequencing of her parents and bioinformatic analysis. RESULTS: The child, an 11-year-old female, had a complain of "scoliosis found 8 years before and aggravated with unequal shoulder height for 1 year". WES results revealed that she has carried a homozygous c.55+1G>C splice variant of the CHRNG gene, for which both of her parents were heterozygous carriers. By bioinformatic analysis, the c.55+1G>C variant has not been recorded by the CNKI, Wanfang data knowledge service platform and HGMG databases. Analysis with Multain online software suggested that the amino acid encoded by this site is highly conserved among various species. As predicted with the CRYP-SKIP online software, the probability of activation and skipping of the potential splice site in exon 1 caused by this variant is 0.30 and 0.70, respectively. The child was diagnosed with MPS. CONCLUSION The CHRNG gene c.55+1G>C variant probably underlay the MPS in this patient.
Humans ; Child ; Female ; Abnormalities, Multiple/genetics* ; Malignant Hyperthermia/genetics* ; Skin Abnormalities/genetics* ; Heterozygote ; Mutation ; Receptors, Nicotinic/genetics*

Epilepsy is a chronic neurological disorder, which is not only related to the imbalance between excitatory glutamic neurons and inhibitory GABAergic neurons, but also related to abnormal central cholinergic regulation. This article summarizes the scientific background and experimental data about cholinergic dysfunction in epilepsy from both cellular and network levels, further discusses the exact role of cholinergic system in epilepsy. In the cellular level, several types of epilepsy are believed to be associated with aberrant metabotropic muscarinic receptors in several different brain areas, while the mutations of ionotropic nicotinic receptors have been reported to result in a specific type of epilepsy-autosomal dominant nocturnal frontal lobe epilepsy. In the network level, cholinergic projection neurons as well as their interaction with other neurons may regulate the development of epilepsy, especially the cholinergic circuit from basal forebrain to hippocampus, while cholinergic local interneurons have not been reported to be associated with epilepsy. With the development of optogenetics and other techniques, dissect and regulate cholinergic related epilepsy circuit has become a hotspot of epilepsy research.
Acetylcholine ; physiology ; Basal Forebrain ; pathology ; Brain Chemistry ; genetics ; physiology ; Cholinergic Neurons ; chemistry ; classification ; pathology ; physiology ; Epilepsy ; genetics ; pathology ; physiopathology ; Epilepsy, Frontal Lobe ; genetics ; GABAergic Neurons ; physiology ; Hippocampus ; pathology ; Humans ; Mutation ; genetics ; physiology ; Neurons ; Non-Neuronal Cholinergic System ; genetics ; physiology ; Receptors, Muscarinic ; genetics ; physiology ; Receptors, Nicotinic ; genetics ; physiology ; Synaptic Transmission ; genetics ; physiology

BACKGROUNDCongenital myasthenic syndromes are a group of rare disorders that are clinically and genetically heterogeneous and caused by mutations in the genes encoding proteins of the neuromuscular junction. Here, we described a Chinese family that presented with phenotypes of classic slow-channel congenital myasthenic syndrome (SCCMS).

METHODSClinical characteristics and electrophysiological features of three patients from a Chinese family were examined, and next-generation sequencing followed by direct sequencing was carried out.

RESULTSThe patients revealed variability in clinical and electrophysiological features. However, weakness, scoliosis, and repetitive-compound muscle action potential were found in all affected members in the family. A heterozygous C>T missense mutation at nucleotide 865 in acetylcholine receptor epsilon-subunit (CHRNE) gene that causes a leucine-to-phenylalanine substitution at position 289 (L289F) was found.

CONCLUSIONSWe reported a SCCMS family of Chinese origin. In the family, classical clinical phenotype with phenotypic variability among different members was found. Genetic testing could help diagnose this rare disease.

Adult ; DNA Mutational Analysis ; Electrophysiology ; Female ; Humans ; Male ; Mutation, Missense ; genetics ; Myasthenic Syndromes, Congenital ; genetics ; physiopathology ; Receptors, Nicotinic ; genetics ; Young Adult

Recent population-based genome wide association studies have revealed potential susceptibility loci of lung cancer at the region of chromosome 15q25.1 containing nicotinic acetylcholine receptor genes. The loci increasing lung cancer risk has been widely identified in Caucasians, but whether this association also exists in Asians and whether this association is a direct role or mediated via tobacco smoking indirectly has not been fully established. We conducted a case-control study comprising of 210 histologically confirmed lung cancer cases and 200 healthy controls to examine rs1051730 genotyping, a single nucleotide polymorphism receiving much attention recently, and its influence on lung cancer risk as well as nicotine dependence in a Chinese Han population. Our results showed that the heterozygous C/T genotype and minor allele T conferred a significant higher risk of lung cancer than the CC homozygotes and allele C (adjusted OR=2.25, 95% CI=1.04-4.89, P=0.040 and OR=2.18, 95% CI=1.02-4.67, P=0.045 respectively). However, no association between the smoking habit and the CHRNA3 rs1051730 polymorphism was observed in this study. The results suggested that the rs1051730 polymorphism may modify susceptibility to lung cancer via a smoking-independent manner among Chinese Han population. Additional studies in vitro and in vivo are warranted to further elucidate the impact of rs1051730 on lung cancer susceptibility.
Adult ; Aged ; Case-Control Studies ; China ; Female ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lung Neoplasms ; etiology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Receptors, Nicotinic ; genetics ; Smoking

OBJECTIVETo investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).

METHODSThe levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.

RESULTSCompared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.

CONCLUSIONThe siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; metabolism ; Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Signal Transduction ; Transfection ; bcl-2-Associated X Protein ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate mutations of CHRNA4 gene in Chinese patients with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE).

METHODSTwo hundred and fifty-seven patients (including 215 sporadic and 42 familial cases) were analyzed. Mutational screening was performed by sequencing all of the 6 exons of the CHRNA4 gene including the donor and acceptor splice sites.

RESULTSThe results have excluded the involvement of any known mutations of the CHRNA4 gene. A novel synonymous mutation c.570C>T(D190D) and 6 single nucleotide polymorphisms (SNPs) of the CHRNA4 gene were detected in 6 sporadic cases, including c.639T/C, c.678T/C, c.1209G/T, c.1227T/C, c.1659G/A, and c.1629C/T. The SNP D190D was hererozygous and absent in 200 healthy controls.

CONCLUSIONThis results suggested that mutations of the CHRNA4 gene may be rare in southern Chinese population with ADNFLE. The synonymous mutation D190D has not been reported previously. Its impact on the pathogenesis of ADNFLE warrant further study.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; methods ; Epilepsy, Frontal Lobe ; genetics ; Female ; Genes, Dominant ; Humans ; Infant ; Male ; Mutation ; Pedigree ; Polymorphism, Single Nucleotide ; Receptors, Nicotinic ; genetics ; Young Adult

OBJECTIVETo investigate the influence of inhibited α7 neuronal nicotinic acetylcholine receptor (nAChR) by small interference RNA (siRNA) in SH-SY5Y cells and to explore the connection of these changes with the β-amyloid precursor protein (APP) metabolism and the pathogenesis of Alzheimer's disease (AD).

METHODSThe siRNA of α7 nAChR was transfected into SH-SY5Y cells, and the expression of α7 nAChR and two subtypes of β-secretases (BACE1 and BACE2) at mRNA and protein levels was studied by real-time PCR and Western blot, respectively. The variation of Aβ(1-42) content was detected by ELISA.

RESULTSAs compared with controls, the expression of α7 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with the α7 nAChR siRNA were decreased by 84% and 79% (P < 0.01), respectively. The expressions of BACE1 mRNA and protein levels was increased by 527% and 71% (P < 0.01), respectively, while the expression of BACE2 decreased by 58% and 75% (P < 0.01), respectively. The Aβ(1-42) content increased by 208% (P < 0.01).

CONCLUSIONSAn inhibited α7 nAChR mRNA induced by siRNA may markedly stimulate the production of Aβ through the mechanism of increased expression of BACE1 and inhibited expression of BACE2, which may be related to the pathogenesis of AD.

Amyloid Precursor Protein Secretases ; genetics ; metabolism ; Amyloid beta-Peptides ; metabolism ; Aspartic Acid Endopeptidases ; genetics ; metabolism ; Cell Line, Tumor ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Transfection ; alpha7 Nicotinic Acetylcholine Receptor

A genome-wide association study has identified the 15q25 region as being associated with the risk of chronic obstructive pulmonary disease (COPD) in Caucasians. This study intended as a confirmatory assessment of this association in a Korean population. The rs6495309C > T polymorphism in the promoter of nicotinic acetylcholine receptor alpha subunit 3 (CHRNA3) gene was investigated in a case-control study that consisted of 406 patients with COPD and 394 healthy control subjects. The rs6495309 CT or TT genotype was associated with a significantly decreased risk of COPD when compared to the rs6495309 CC genotype (adjusted odds ratio = 0.69, 95% confidence interval = 0.50-0.95, P = 0.023). The effect of the rs6495309C > T on the risk of COPD was more evident in moderate to very severe COPD than in mild COPD under a dominant model for the variant T allele (P = 0.024 for homogeneity). The CHRNA3 rs6495309C > T polymorphism on chromosome 15q25 is associated with the risk of COPD in a Korean population.
Adult ; Aged ; Alleles ; Asian Continental Ancestry Group/*genetics ; Case-Control Studies ; Female ; Forced Expiratory Volume ; Genotype ; Humans ; Male ; Middle Aged ; Odds Ratio ; *Polymorphism, Single Nucleotide ; Pulmonary Disease, Chronic Obstructive/*genetics/physiopathology ; Receptors, Nicotinic/*genetics ; Republic of Korea ; Risk Factors ; Smoking

OBJECTIVETo observe the effect of Scutellarin (Scu) on expressions of nicotinic acetylcholine receptor (nAChR) subunit protein and mRNA in dementia rats, and to study its possible mechanism on dementia.

METHODSForty-two Wistar rats were randomly divided into 5 groups, i.e., the normal control group (n=6), the sham-operative group (n=6), the memory deficit model group, the Scu treatment group (n=10), and the positive drug (piracetam) control group (n=10). The dementia rat model was established by bilateral ventricle injection with beta-amyloid peptide (Abeta)(25-35) and abdominal cavity injection with D-galactose. Rats in the Scu treatment group or the piracetam control group were treated with Scu or piracetam by gastrogavage. The learning and memory ability of rats were detected by Morris water maze test, nAChR alpha4, alpha7, and beta2 subunits at protein and mRNA levels were detected by Western blot and Real-time PCR respectively.

RESULTSCompared with the normal control group and the sham-operative group, the learning and memory ability decreased in rats of the model group (P<0.05). nAChR alpha4 and alpha7 subunit protein expressions were obviously lowered (P<0.05), but changes of beta2 were not obvious. No obvious change of mRNA expressions in all three nAChR subunits was seen (P>0.05). After treatment of Scu, the learning and memory ability was greatly improved, nAChRs alpha4 and alpha7 subunit protein expressions increased in rats with dementia (all P<0.05). No obvious change of mRNA expressions in all three nAChR subunits was seen (P>0.05). No obvious difference of each index was shown between the Scu treatment group and the positive drug (piracetam) control group.

CONCLUSIONSScutellarin could improve the learning and memory ability of dementia rats. Its mechanism might be associated with its up-regulation of nAChR expressions.

Alzheimer Disease ; drug therapy ; genetics ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Apigenin ; pharmacology ; therapeutic use ; Brain ; drug effects ; metabolism ; Disease Models, Animal ; Female ; Glucuronates ; pharmacology ; therapeutic use ; Learning ; drug effects ; Male ; Memory ; drug effects ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Nicotinic ; genetics ; metabolism ; Up-Regulation

BACKGROUNDIt has been reported that the nicotinic acetylcholine receptor subunit α4 gene (CHRNA4) might be associated with smoking behaviors in the previous studies. Up to now, there are few reports on the relationship between CHRNA4 and smoking initiation. In this study, we tried to explore the role of two polymorphisms in CHRNA4 (rs1044396 and rs1044397) in smoking initiation and nicotine dependence in Chinese male smokers.

METHODSNine hundred and sixty-six Chinese male lifetime nonsmokers and smokers were assessed by the Fagerström test for nicotine dependence (FTND), smoking quantity (SQ) and the heaviness of smoking index (HSI). All subjects were divided into four groups based on their tobacco use history and the FTND scores. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to find two polymorphisms of CHRNA4 in these subjects.

RESULTSThe χ(2) test showed that rs1044396 was significantly associated with smoking initiation (χ(2) = 4.65, P = 0.031), while both rs1044396 and rs1044397 were significantly associated with nicotine dependence (χ(2) = 5.42, P = 0.020; χ(2) = 7.58, P = 0.005). Furthermore, the T-G (3.9%) haplotype of rs1044396-rs1044397 showed significant association with smoking initiation (χ(2) = 6.30, P = 0.012) and the C-G haplotype (58.9%) remained positive association with nicotine dependence (χ(2) = 8.64, P = 0.003) after Bonferroni correction. The C-G haplotype also significantly increased the HSI (P = 0.002) and FTND scores (P = 0.001) after Bonferroni correction.

CONCLUSIONThese findings suggest that CHRNA4 may be associated with smoking initiation and the C-G haplotype of rs1044396-rs1044397 might increase the vulnerability to nicotine dependence in Chinese male smokers.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics ; Receptors, Nicotinic ; genetics ; Smoking ; adverse effects