Rapid quantification of the heteroplasmy of mutant mitochondrial DNAs in Leber's hereditary optic neuropathy using the Invader technology

PURPOSE: To quantify the degree of heteroplasmy of a mitochondrial DNA (mtDNA) mutation in Leber's hereditary optic neuropathy (LHON) a biplex Invader assay was applied. METHODS: To determine the optimum condition for the Invader assay, mtDNAs were assayed in various amounts of total DNA in 1-4-h incubations at 63 degrees C. To evaluate the suitability of the Invader assay to detect the three mutations, G3460A, G11778A, and T14484C, 10 ng of DNAs from 224 patients with bilateral optic atrophy was assayed. To quantify mtDNA heteroplasmy, a standard curve of known mixture ratios of mutation against calculation by the Invader assay was constructed. Seventy-two of the 224 patients had one of the three mutations, which corresponded with the mutation detected earlier by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. The percentages of mutant mtDNAs were calculated by the Invader assay in five heteroplasmic families, including 30 individuals with the G11778A mutation. The results were compared with those calculated earlier by labeled polymerase chain reaction followed by single-strand conformation polymorphism (PCR-SSCP) analysis. RESULTS: In 1-8 ng of DNA, the fluorescence intensity increased near linearly during a 4-h assay. With more than 16 ng of DNA, the intensities were saturated even at the 2-h assay. A linear relationship was observed between the results obtained from separate mixtures and from the Invader assay analysis. Because two fluorescent intensities are not always the same, one of the two intensities was modified to adjust to that of the other. Complete concordance was observed between PCR-RFLP analysis and Invader assay genotyping for the 224 patients. Results of percentage of heteroplasmy in five LHON families obtained by the Invader assay were consistent with those by the PCR-SSCP analysis. CONCLUSIONS: Invader assay is a simple, rapid, and reliable method of genotyping mtDNA mutations as well as quantifying heteroplasmy simultaneously under optimum conditions.     

应用HRM分析Leber遗传性视神经病变家系mtDNA突变

目的揭示一个Leber遗传性视神经病变（Leber＇s hereditary optic neuropathy,LHON）家系的遗传基础。方法对家系成员提取线粒体DNA（mtDNA）,直接进行测序分析,用分子克隆法为每个人构建单克隆群,再用高分辨熔解曲线技术和DNA测序的方法,对家系中成员的单克隆群分析统计,计算该家系成员的线粒体DNA突变比例。结果该家系患者mtDNA上11778位核苷酸发生G到A的突变。家系成员中G11778A突变比例分别为：先证者（112）91．67％;父亲（12）0％;3位母系家属正常人依次为：（11）90．83％、（111）53．16％、（113）49．16％。结论G11778A的同质体（即：突变比例达到90％以上）女性仍可不患Leber遗传性视神经病变,该女性后代的平均突变比例远小于先前的报道。     

Efficacy of levodopa and carbidopa on visual function in patients with non-arteritic anterior ischaemic optic neuropathy

The benefit of levodopa and carbidopa therapy in improving visual function in patients with non-arteritic anterior ischaemic neuropathy (NAION) was evaluated. Twenty-four subjects with NAION were randomly selected to receive either levodopa-carbidopa or a placebo. Visual functions of neither the study nor the placebo groups showed improvement. Side effects of levodopa such as dizziness, orthostatic hypotension, vomiting and cardiac arrhythmia were seen. Levodopa and carbidopa had no therapeutic effect on visual recovery in our patients with NAION.     

视神经减压术对颅脑损伤合并视神经损伤视力康复的意义

Therewasmuchcontroversyovertheeffectofsurgicaltreatmentonopticneuropathyduetoheadtrauma．From１９９５～２００１，wecomparedtheeffectofdecompressionofopticnerveandconservativetreatmentonopticneuropathyfollowingheadtraumareceivedin１６patients．Therapeuticeffectofdecompressionissatisfying，hereisthereport．１Subjectandmethod１．１Subject１６patientsincluded１４menand２womenagedbetween１６～４５yearsoldwithmeanageof２９．５years．Allheadtraumaswerecausedbytrafficaccident．５casessufferedfromuni…     

经颅磁刺激运动诱发电位和体感诱发电位对遗传性运动感觉性神经病的诊断

目的了解遗传性运动感觉性神经病(HMSN)的经颅磁刺激运动诱发电位(MEP)和体感诱发电位(SEP)的变化。方法对一家三代HMSN的12例患者和1例无症状者进行检查。结果MEP和SEP的异常率分别为84.6%,92.3%。结论绝大多数HMSN患者的MEP和SEP均异常。     

Nonarteritic anterior ischemic optic neuropathy: associations with homozygosity for the C677T methylenetetrahydrofolate reductase mutation

The association between nonarteritic anterior ischemic optic neuropathy (NAION) and coagulation disorders was prospectively assessed at least 3 months after the occurrence of ocular vascular events in 12 white patients in an outpatient clinical research center. Two community-based ophthalmologists evaluated the 12 NAION patients in the consecutive order of their referral. Polymerase chain reaction-complementary DNA assays of gene mutations associated with coagulation disorders and serologic coagulation measurements in study patients were compared with those in 36 healthy, normal race-, sex-, and age-matched controls, with 3 controls matched for each case. Of the 12 patients, 4 men and 8 women (mean age 62 +/- 15 years, 3 of them 55 years or older), 8 had unilateral NAION (bilateral in 4). The 12 patients with NAION were more likely than controls to demonstrate homozygosity for the methylenetetrahydrofolate reductase (MTHFR) C677T mutation (50% vs 11 %; Fisher's P =.009, with the likelihood of a type I error quite small, 0.9%). Our sample size had a power of 80% to detect this case-control difference in C677T MTHFR homozygosity at an alpha value of.05. Of the 12 NAION patients, 7 (58%) had at least 1 gene mutation in the C677T MTHFR, G1691A V Leiden, or G20210A prothrombin gene, compared with 5 of 36 controls (14%) (chi(2) = 9.48, P =. 002, with the likelihood of a type I error quite small, 0.2%). Our sample size had a power of 85% to detect this case-control difference at alpha =. 05. Of the 8 women with NAION, 5 (63%) first experienced the condition while taking hormone replacement therapy (n = 4) or during pregnancy (n = 1), with superposition of estrogen-induced thrombophilia on heritable thrombophilia and hypofibrinolysis. Confirmation of a causal relationship between coagulation disorders and NAION should facilitate its prevention and treatment and help protect against thrombi in other vascular beds.     

The identification of a novel frameshift insertion mutation in the EXT1 gene in a Chinese family with hereditary multiple exostoses

To identify the pathogenic gene variation in a Chinese family with Hereditary Multiple Exostoses (HME). By examining blood‐sourced DNA and clinical manifestations of the proband and his family members, the whole exome sequencing (WES) and Sanger sequencing were used to detect possibly pathogenic mutations. A novel heterozygous mutation (c.325dup) was identified in exon 1 of the exostosin 1 (EXT1) gene from the proband and the affected family members. And we found this mutation was absent in all the unaffected family members. This c.325dup mutation is in the exon 1 domain of the EXT1 gene and the change of p.C109Lfs*80 cause the early termination of protein translation. The identification of the novel frameshift insertion mutation (c.325dup) expands the mutation spectrum of HME, which provides new evidence for HME diagnosis.     

The characteristics of upper airway edema in hereditary and acquired angioedema with C1‐inhibitor deficiency

BackgroundAngioedemas localized in the upper airway are potentially life threatening, and without proper treatment, they may lead to death by suffocation. Upper airway edemas (UAE) in bradykinin‐mediated angioedemas can even be the first symptoms of the disease.MethodsOur survey was performed with a retrospective long‐term follow‐up method from the medical history of 197 hereditary (C1‐INH‐HAE) and 20 acquired C1‐inhibitor deficiency (C1‐INH‐AAE), 3 factor XII and 3 plasminogen gene mutation (FXII‐HAE, PLG‐HAE) patients treated at our center between 1990 and 2020. The UAE group included edemas localized to the mesopharynx, hypopharynx, and larynx, as narrowing of these anatomical regions can lead to suffocation.Results98/197 C1‐INH‐HAE (47 families) and 13/20 C1‐INH‐AAE, 1/3 PLG‐HAE, 1/3 FXII‐HAE patients had experienced UAE at least once according to their medical history. In case of C1‐INH‐HAE patients, in 6/47 families who had undiagnosed ancestors had 13 members who died of suffocation. After the diagnosis, 1‐1 member of two families died of UAE. 44/64 C1‐INH‐HAE patients did not smoke, 20/64 did. The occurrence of UAE was significantly higher in smoker patients. We analyzed 7607 HAE attacks of 56/98 patients. Out of all attacks, the incidence of UAE in the C1‐INH‐HAE group was 4%, and 9.5% in the C1‐INH‐AAE group, respectively.ConclusionEarly diagnosis is key in bradykinin‐mediated angioedemas cases, since the patient must be provided with adequate treatment; and also it is essential to inform patients about the importance of avoiding the trigger factors and the early symptoms of UAE, as these measures could significantly decrease the incidence of lethal UAEs.     

The antinociceptive effect and adverse drug reactions of oxycodone in human experimental pain in relation to genetic variations in the OPRM1 and ABCB1 genes

The aim of this study was to search for a possible association between the variant allele of the single nucleotide polymorphisms A118G in the OPRM1 gene and C3435T and G2677T/A in the ABCB1 gene and altered antinociceptive effect and adverse drug reactions of oxycodone. Thirty‐three healthy subjects exposed to experimental pain including electrical stimulation and the cold pressor test were included. A118G: We found that the variant G allele was associated with reduced antinociceptive effect as measured by pain tolerance thresholds to single electrical nerve stimulation (8% increase vs. 25% for the wild‐type carriers, P = 0.007). C3435T: The carriers of the variant T allele generally had less adverse drug reactions on oxycodone than the carriers of the wild‐type genotype. G2677T/A: The carriers of the variant T allele had a better antinociceptive effect of oxycodone than the carriers of the wild‐type genotype in the cold pressor test (25% reduction vs. 15%, P = 0.015 in the discomfort rating and 25% reduction vs. 12%, P = 0.007 in the pain time AUC) and less adverse drug reactions. The combined wild‐type genotype 3435CC‐2677GG was associated with less antinociceptive effect of oxycodone in the discomfort rating of the cold pressor test (13% reduction vs. 23%, P = 0.019) and more severe adverse drug reactions than the carriers of the variant alleles. We found a moderate association between less antinociceptive effect of oxycodone and the variant allele of A118G. There was strong association between less adverse drug reactions of oxycodone and the variant alleles of C3435T and G2677T/A.     

Association between ABO blood group and risk of Crohn’s disease: A case‐control study in the Chinese Han population

BackgroundBlood group O has been reported to be a potentially protective factor for Crohn''s disease (CD) susceptibility in Caucasian and Korean populations, but a similar conclusion was not found in a Chinese study. The present study investigated the potential association in the Chinese Han population.MethodsWe included 275 CD patients, 132 ulcerative colitis (UC) patients and 1201 healthy individuals in this case‐control study. The demographic characteristics and ABO blood group were compared among the three groups. The clinical characteristics and treatment of CD were further investigated according to the blood group distribution.ResultsThe blood group distribution in CD patients was significantly different from healthy controls, and the frequency of O blood in CD patients was significantly lower compared to healthy controls. After adjusting for age and gender, the non‐O blood groups remained significantly associated with CD susceptibility in propensity score‐adjusted and propensity score‐matched analyses. Compared to CD patients with non‐O blood groups, patients with O blood were at a lower risk of developing penetrating disease, more likely to receive immunosuppressant treatment and less likely to receive biological treatment.ConclusionOur results confirmed that non‐O blood groups were significantly associated with an increased risk of CD in the Chinese Han population.     

Evaluation of Optic Nerve with Strain and Shear Wave Elastography in Patients with Behçet's Disease and Healthy Subjects

The objective of this study was to investigate the elasticity characteristics of the optic nerve using strain and shear wave elastography in patients with Behçet's disease and to compare the results with those of healthy volunteers. Forty-six optic nerves from patients with Behçet's disease and 54 optic nerves from healthy volunteers were investigated prospectively in this study using strain and shear wave elastography. There was a statistically significant difference in terms of elasticity patterns between patients and healthy volunteers (p < 0.001). Elastographic images of healthy volunteers revealed most optic nerves to be type 3 (51.8%); however, type 2 (40.7%) and type 1 (7.5%) were also observed. Elastographic examination of Behçet's disease patients revealed type 2 in 52.2%, type 1 in 43.5% and type 3 in 4.3% of patients. Statistically significant differences were observed between patients and healthy volunteers in the analysis of shear wave elastography values (p < 0.001). Receiver operating characteristic curve analysis was perfect (0.933) (95% CI = 0.885–0.980), and a cutoff value of 16.5 kPa shear had very high sensitivity and specificity for the patient group. Strain and shear wave elastography findings for the optic nerves of patients with Behçet's disease were significantly different from those for healthy volunteers.     

佛山地区孕产妇G6PD基因缺陷调查及干预模式研究

目的调查佛山地区孕产妇葡萄糖-6-磷酸脱氢酶(G6PD)缺陷状况,探讨干预指导模式的效果。方法选择2013年4~12月到该院产检的孕产妇及其丈夫各1 646例,以反向膜杂交法检查夫妇双方的G6PD基因、以生化仪法检测G6PD活性,分析两性基因突变率、酶活性缺陷率,给予遗传咨询指导并跟踪随访新生儿G6PD基因突变与酶活性低下情况。结果 G6PD基因突变率男性4.7%,女性2.3%;G6PD酶活性缺陷率男性3.5%,女性1.6%;G6PD基因突变发生较高的位点是G1376T(36.2%)、G1388A(21.6%)、A95G(13.7%)和G1024T(11.2%)。结论佛山地区是G6PD缺陷高发地区,G1376T、G1388A、A95G和G1024T是主要的点突变类型。