Detection of mtDNA with 4977 bp deletion in blood cells and atherosclerotic lesions of patients with coronary artery disease

Recent evidence suggests that somatic mutations in nuclear and mitochondrial DNA accumulated during aging, may significantly contribute to the pathogenesis of chronic-degenerative illness such as coronary artery disease (CAD). Mitochondrial DNA with 4977 bp deletion mutation (mtDNA4977) is a common type of mtDNA alteration in humans. However, little attempt has been made to detect the presence of mtDNA4977 deletion in cells and tissues of cardiovascular patients. This study investigated the presence of mtDNA4977 in blood samples of 65 cardiovascular patients and 23 atherosclerotic plaques of human coronaries with severe atherosclerosis. Moreover, the presence of the deletion has been investigated in blood cells from 22 healthy age-matched subjects. The detection of mtDNA4977 has been performed by using a nested polymerase chain reaction (PCR) protocol and normalized to wild-type mtDNA. A significant higher incidence of mtDNA4977 was observed in CAD patients with respect to healthy subjects (26.2% versus 4.5%; P=0.03). Furthermore, the relative amount of the deletion was significantly higher in the patients compared to the control group (P=0.02). The mtDNA4977 was detected in 17 of the 65 patients blood samples (26.2%) and deletion levels ranged from 0.18 to 0.46% of the total mtDNA (mean: 0.34+/-0.02%). For what concerns atherosclerotic lesions, 5 patients (21.7%) showed the deletion ranging from 0.13 to 0.45% of the total mtDNA (mean: 0.35+/-0.06%). In both samples from patients, the incidence and the relative amount of mtDNA4977 was not significantly influenced by atherogenic risk factors and clinical parameters. The obtained results may suggest that the increase of oxidative stress in cardiovascular disease may be responsible for the accumulation of mtDNA damage in coronary artery disease patients.     

The discovery and phylogenetic implications of a novel 41 bp plastid DNA deletion in wild potatoes

Insertions and deletions (indels) are common in intergenic spacer regions of plastid DNA and can provide important phylogenetic characters for closely related species. For example, a 241-bp plastid DNA deletion in the trnV-UAC/ndhC intergenic spacer region has been shown to have major phylogenetic importance in determining the origin of the cultivated potato. As part of a phylogenetic study of the wild potato Solanum series Piurana group we screened 199 accessions of 38 wild potato species in nine of the 19 tuber-bearing (Solanum section Petota) series that have not been examined before for indels in the trnV-UAC/ndhC intergenic spacer region. A novel 41 bp deletion (but no 241 bp deletion) was discovered for 30 accessions of three species: S. chiquidenum (5 of 10 accessions), S. chomatophilum (19 of 28), and S. jalcae (6 of 6). Accessions with and without this deletion are found throughout much of the north-south range of all three species in northern and central Peru, but not east of the Marañón River. Multivariate morphological analyses of these 44 accessions showed no morphological associations to the deletion. The results suggest extensive interspecific gene flow among these three species, or a common evolutionary history among species that have never been suggested to be interrelated.     

Comparison of the mitochondrial genome of Nicotiana tabacum with its progenitor species

Summary Mitochondrial DNAs from Nicotiana tabacum, an amphiploid, and its putative progenitor species, N. sylvestris and N. tomentosiformis were compared in structure and organization. By using DNA transfer techniques and cloned fragments of known genes from maize and N. sylvestris as labeled probes, the positions of homologous sequences in restriction digests of the Nicotiana species were analyzed. Results indicate that the mitochondrial DNA of N. tabacum was inherited from N. sylvestris. Conservation in organization and sequence homology between mtDNAs of N. tabacum and the maternal progenitor, N. sylvestris, provide evidence that the mitochondrial genome in these species is evolutionarily stable. Approximately one-third of the probed restriction fragments of N. tomentosiformis mtDNA showed conservation of position with the other two species. Pattern variations indicate that extensive rearrangement of mtDNA has occurred in the evolution of these Nicotiana species.     

原发性高血压合并冠心病患者血清CRP的变化

目的:探讨原发性高血压合并不同类型冠心病患者血清C-反应蛋白(CRP)的变化及其意义。方法:用全自动生化分析仪检测71例原发性高血压患者和28例健康对照组人群的血清CRP水平,所有研究对象均行选择性冠状动脉造影检查。将原发性高血压组病人根据冠脉造影检查分为单纯原发性高血压组(EH),原发性高血压合并稳定性心绞痛组(EH+SAP)、原发性高血压合并不稳定心绞痛组(EH+UAP)、原发性高血压合并心肌梗死组(EH+MI),比较各组患者血清CRP水平的差异。结果:①与对照组比较,EH组及EH合并CAD各组患者血清CRP均显著升高(P0.05);②与EH组比较,EH+UAP和EH+MI组血清CRP水平均显著升高(P0.01);③与EH+SAP组比较,EH+AMI组和EH+UAP组血清CRP水平均显著升高(P0.01)。结论:冠心病合并原发性高血压患者血CRP水平与冠状动脉病变程度与斑块稳定程度存在正相关性。     

The mitochondrial ND1 T3308C mutation in a Chinese family with the secondary hypertension

Mutations in mitochondrial DNA have been associated with hypertension. We report here the clinical, genetic, and molecular characterization of one four-generation Han Chinese family with hypertension. Two matrilineal relatives in this family exhibited the variable degree of a secondary hypertension (renal hypertension) at the age-at-onset of 42 and 56 years old, respectively. Sequence analysis of the complete mitochondrial DNA in this pedigree revealed the presence of the known hypertension-associated ND1 T3308C mutation and 42 other variants, belonging to the Asian haplogroup D4h. The T3308C mutation resulted in the replacement of the first amino acid, translation-initiating methionine with a threonine in ND1. Furthermore, the ND3 T3308C mutation also locates in two nucleotides adjacent to the 3′ end of mitochondrial tRNA^Leu(UUR). Thus, this T3308C mutation caused an alteration on the processing of the H-strand polycistronic RNA precursors or the destabilization of ND1 mRNA. The occurrence of the T3308C mutation in these genetically unrelated pedigrees affected by diseases but absence of 242 Chinese controls as well as the mitochondrial dysfunctions detected in cells carrying this mutation indicate that this mutation is involved in the pathogenesis of hypertension. However, the mild biochemical defects, the lower penetrance of hypertension in this Chinese family and the presence of some control populations suggested the involvement of other modifier factors in the pathogenesis of hypertension associated with this ND1 T3308C mutation.     

High‐altitude adaptation in vertebrates as revealed by mitochondrial genome analyses

The high‐altitude environment may drive vertebrate evolution in a certain way, and vertebrates living in different altitude environments might have different energy requirements. We hypothesized that the high‐altitude environment might impose different influences on vertebrate mitochondrial genomes (mtDNA). We used selection pressure analyses and PIC (phylogenetic independent contrasts) analysis to detect the evolutionary rate of vertebrate mtDNA protein‐coding genes (PCGs) from different altitudes. The results showed that the ratio of nonsynonymous/synonymous substitutions (dN/dS) in the mtDNA PCGs was significantly higher in high‐altitude vertebrates than in low‐altitude vertebrates. The seven rapidly evolving genes were shared by the high‐altitude vertebrates, and only one positive selection gene (ND5 gene) was detected in the high‐altitude vertebrates. Our results suggest the mtDNA evolutionary rate in high‐altitude vertebrates was higher than in low‐altitude vertebrates as their evolution requires more energy in a high‐altitude environment. Our study demonstrates the high‐altitude environment (low atmospheric O₂ levels) drives vertebrate evolution in mtDNA PCGs.     

Maternally inherited hypertension is associated with the mitochondrial tRNA(Ile) A4295G mutation in a Chinese family

Mutations in mitochondrial DNA have been associated with cardiovascular disease. We report here the clinical, genetic, and molecular characterization of one three-generation Han Chinese family with maternally transmitted hypertension. All matrilineal relatives in this family exhibited the variable degree of hypertension at the age at onset of 36 to 56 years old. Sequence analysis of the complete mitochondrial DNA in this pedigree revealed the presence of the known hypertension-associated tRNA^Ile A4295G mutation and 33 other variants, belonging to the Asian haplogroup D4j. The A4295G mutation, which is extraordinarily conserved from bacteria to human mitochondria, is located at immediately 3′ end to the anticodon, corresponding to conventional position 37 of tRNA^Ile. The occurrence of the A4295G mutation in several genetically unrelated pedigrees affected by cardiovascular disease but the absence of 242 Chinese controls strongly indicates that this mutation is involved in the pathogenesis of cardiovascular disease. Of other variants, the tRNA^Glu A14693G and ND1 G11696A mutations were implicated to be associated with other mitochondrial disorders. The A14693G mutation, which is a highly conserved nucleoside at the TψC-loop of tRNA^Glu, has been implicated to be important for tRNA structure and function. Furthermore, the ND4 G11696A mutation was associated with Leber’s hereditary optic neuropathy. Therefore, the combination of the A4295G mutation in the tRNA^Ile gene with the ND4 G11696A mutation and tRNA^Glu A14693G mutation may contribute to the high penetrance of hypertension in this Chinese family.     

2型糖尿病患者血浆chemerin水平与其冠脉病变程度的相关性分析

目的:探讨2型糖尿病患者血浆趋化素(chemerin)水平与冠脉病变程度的关系。方法:选取2011年1月~2012年2月我院收治的2型糖尿病(T2DM)患者92例,均行冠状动脉造影检查。按冠状动脉造影结果分为单纯2型糖尿病无冠状动脉病变组26例(DM0);合并单支冠状动脉病变组32例(DM1);合并双支以上病变组34例(DM2)。另选取正常健康行冠脉造影检查者25例作为正常对照组(NC)。采用酶联免疫吸附法检测各组入选者血浆chemerin水平,并分析其与冠脉病变程度的相关性。结果:T2DM及T2DM合并冠状动脉病变患者血浆chemerin水平与NC组对比均明显升高,并且与冠状动脉病变严重程度呈正相关(P0.05)。各组血浆Chemerin水平与BMI、HOMA-IR、TC、和APOB各指标间呈显著正相关(r分别为0.781、0.723、0.415、0.694,P均0.01)。结论:2型糖尿病患者冠状动脉病变的发生发展可能与血浆chemerin升高有关。     

老年2型糖尿病患者血清促甲状腺激素水平及其与冠脉病变程度的相关性分析

目的:探讨老年2型糖尿病(T2DM)患者血清促甲状腺激素(TSH)水平及其与患者冠脉病变程度的相关性。方法:回顾性分析2015年6月～2017年6月我院收治的88例老年T2DM患者(T2DM组)、50例健康体检者(对照组)的临床资料,根据血清TSH水平将T2DM组分为四个亚组,A组(0.45～1.49 m IU/L,n=18)、B组(1.50～2.49 m IU/L,n=23)、C组(2.50～3.49 m IU/L,n=22)、D组(≥4.5 m IU/L,n=25)。比较T2DM组与对照组TSH水平的差异,并根据计算机断层血管造影(CTA)结果计算Gensini评分,分析Gensini评分与血清TSH水平的相关性。结果:T2DM组血清TSH水平显著高于对照组,且随着血清TSH水平的升高,T2DM患者的年龄、TC、TG、LDL-C明显增加,而HDL-C、T3明显降低(P0.05)。C组病变支数显著多于A组,重度病变的比例明显升高,而D组病变支数、病变程度与A组、B组、C组比较差异均有统计学意义(P0.05)。血清TSH水平与Gensini评分呈显著正相关(r=0.577,P0.05)。结论:老年T2DM患者血清TSH水平显著升高,且与患者冠脉病变严重程度呈显著正相关,血清TSH水平有助于评估老年T2DM患者冠脉病变的严重程度。     

血清CTRP4、CTRP5、CTRP6与2型糖尿病合并冠心病患者冠状动脉病变的关系研究

摘要 目的：探讨血清补体1q/肿瘤坏死因子相关蛋白（CTRP）4、CTRP5、CTRP6与2型糖尿病（T2DM）合并冠心病（CHD）患者冠状动脉病变的关系。方法：选取2021年3月～2021年7月我院收治的100例T2DM合并CHD患者为合并组,根据SYNTAX评分分为中重度病变组44例和轻度病变组56例,选取同期收治的100例单纯T2DM患者为T2DM组,另选取同期体检健康志愿者80名为对照组。采用酶联免疫吸附法检测血清CTRP4、CTRP5、CTRP6水平。采用Spearman相关性分析T2DM合并CHD患者SYNTAX评分与血清CTRP4、CTRP5、CTRP6水平的相关性,多因素Logistic回归分析T2DM合并CHD患者冠状动脉中重度病变的影响因素。结果：对照组、T2DM组、合并组血清CTRP4、CTRP5水平依次升高,CTRP6水平依次降低（P＜0.05）。Spearman相关性分析显示,T2DM合并CHD患者SYNTAX评分与血清CTRP4、CTRP5水平呈正相关（rs=0.820、0.833,P均＜0.001）,与CTRP6水平呈负相关（rs=-0.838,P＜0.001）。多因素Logistic回归分析显示,T2DM病程延长和糖化血红白蛋白（HbA1c）、CTRP4、CTRP5升高为T2DM合并CHD患者冠状动脉中重度病变的独立危险因素,CTRP6升高为独立保护因素（P＜0.05）。结论：血清CTRP4、CTRP5水平升高和CTRP6水平降低与T2DM合并CHD患者冠状动脉病变加重独立相关,可能成为T2DM合并CHD患者冠状动脉病变评估指标。     

Phylogenetic relationships of Hynobius naevius (Amphibia: Caudata) as revealed by mitochondrial 12S and 16S rRNA genes

Using mitochondrial 12S and 16S rRNA sequences, we investigated phylogenetic relationships among populations of the endemic Japanese salamander Hynobius naevius. Monophyly of this species was recovered only in the maximum parsimony tree and was unresolved in maximum likelihood and Bayesian trees. Instead the following four haplotype clades consistently emerged clearly: Clade 1 from northwestern Kyushu, Clade 2 from Chugoku and northeastern Kyushu, Clade 3 from western Shikoku and Kyushu, and Clade 4 from Chubu-Kinki and central-eastern Shikoku. Of these, Clades 1 and 2, and Clades 3 and 4, respectively, correspond to Groups A and B previously recognized from the analyses of allozyme data in this species, but monophyly of these groups was not strongly supported. Unlike the previous results, the western and eastern samples from Shikoku did not form a clade, and were grouped with Kyushu-B in Clade 3 and Chubu-Kinki in Clade 4, respectively. The reason for this conflict between mtDNA and allozyme results is unknown, but might be related to retention of ancestral mtDNA polymorphism in Shikoku populations. Nearly simultaneous divergence of as many as four lineages in wide-ranging H. naevius is inferred for the late Miocene-Pliocene history of this taxon.     

冠状动脉CTA联合血清HCY、Cys-C、ApoB/ApoA1比值对2型糖尿病患者合并冠状动脉病变的诊断价值

摘要 目的：探究冠状动脉CT血管造影（CTA）联合血清同型半胱氨酸（HCY）、胱抑素C（Cys-C）、载脂蛋白B/载脂蛋白A1（ApoB/ApoA1）比值对2型糖尿病（T2DM）患者合并冠状动脉病变的诊断价值。方法：回顾性选取2018年8月到2021年8月间我院收治的358例T2DM患者,均行常规生化指标、CTA检查、冠状动脉造影（CAG）检查,根据CAG检查结果为金标准将T2DM患者分为未合并冠脉病变组（190例）和合并冠脉病变组（168例）,比较两组血清HCY、Cys-C、ApoB/ApoA1比值,分析CTA与CAG诊断冠脉狭窄结果的一致性,应用受试者工作特征（ROC）曲线评估冠状动脉CTA联合血清HCY、Cys-C、ApoB/ApoA1比值对T2DM合并冠状动脉病变的诊断价值。结果：与未合并冠脉病变组比较,合并冠脉病变组血清HCY、Cys-C、ApoB、ApoB/ApoA1比值水平明显更高（P＜0.05）,ApoA1明显更低（P＜0.05）。以CAG为金标准,CTA诊断冠脉狭窄程度与CAG一致性较高（Kappa值0.748）。ROC曲线评估冠状动脉CTA诊断T2DM合并冠脉病变的AUC、灵敏度、特异度、准确度依次为0.802、74.40%、83.71%、79.11%。三项血清指标联合AUC、准确度显著优于单一指标（P＜0.05）。冠状动脉CTA联合血清HCY、Cys-C、ApoB/ApoA1比值诊断T2DM合并冠脉病变的价值显著优于各项指标单一诊断或三项血清指标联合诊断（P＜0.05）。结论：冠状动脉CTA联合血清HCY、Cys-C、ApoB/ApoA1比值诊断T2DM患者合并冠状动脉病变的价值较高,相较各项指标单一应用而言更具优势。     

Changes in energy status of leaf cells as a consequence of mitochondrial genome rearrangement

The MSC16 cucumber (Cucumis sativus L.) mutant with lower activity of mitochondrial Complex I was used to study the influence of mitochondrial metabolism on whole cell energy and redox state. Mutant plants had lower content of adenylates and NADP(H) whereas the NAD(H) pool was similar as in wild type. Subcellular compartmentation of adenylates and pyridine nucleotides were studied using the method of rapid fractionation of protoplasts. The data obtained demonstrate that dysfunction of mitochondrial respiratory chain decreased the chloroplastic ATP pool. No differences in NAD(H) pools in subcellular fractions of mutated plants were observed; however, the cytosolic fraction was highly reduced whereas the mitochondrial fraction was more oxidized in MSC16, as compared to WTc. The NADP(H) pool in MSC16 protoplasts was greatly decreased and the chloroplastic NADP(H) pool was more reduced, whereas the extrachloroplastic pool was much more oxidized, than in WTc protoplast. Changes in nucleotides distribution in cucumber MSC16 mutant were compared to changes found in tobacco (Nicotiana sylvestris) CMS II mitochondrial mutant. In contrast to MSC16 cucumber, the content of adenylates in tobacco mutant was much higher than in tobacco wild type. The differences were more pronounced in leaf tissue collected after darkness than in the middle of the photoperiod. Results obtained after tobacco protoplast fractionating showed that the increase in CMS II adenylate content was mainly due to a higher level in extrachloroplast fraction. Both mutations have a negative effect on plant growth through perturbation of chloroplast/mitochondrial interactions.     

尼可地尔对冠心病合并2型糖尿病不完全血运重建术后的疗效观察

目的:比较尼可地尔与单硝酸异山梨酯对冠心病合并2型糖尿病患者不完全血运重建术后的疗效。方法:入选112例经皮冠状动脉介入治疗(PCI)部分血运重建的冠心病合并2型糖尿病患者。随机分为2组:尼可地尔组(5 mg,3次/d,口服)60例,单硝酸异山梨酯组(50 mg,1次/d,口服)52例,两组患者均给予常规冠心病及糖尿病药物治疗。术后4周末行运动负荷试验,观察总运动时间,从开始运动到出现ST段压低1.0 mv和出现心绞痛的时间(s),以及最大ST段压低幅度,同时记录每周心绞痛发作次数及硝酸甘油用量。结果:4周后两组患者同服药前比较,从开始运动到出现ST段压低1.0 mv的时间延长,总运动时间延长,从开始运动到出现心绞痛的时间延长,最大ST段压低幅度降低(P0.01),但尼可地尔组与单硝酸异山梨酯组比较无明显统计学差异(P0.05);尼可地尔组每周心绞痛发作次数及硝酸甘油用量明显少于单硝酸异山梨酯组(P0.05)。结论:尼可地尔可增加冠心病合并糖尿病患者行不完全血运重建术后患者运动耐量,在减少心绞痛方面作用优于单硝酸异山梨酯。     

Analysis of nucleotide substitutions and amino acid conservation in theDrosophila Adh genomic region

The homologous genomic region that contains two paralogous genes,Adh andAdh-dup, was compared in severalDrosophila species. Sequences were analyzed as follows: a) At the nucleotide level, Ka and Ks values were determined for each pair of species. Ka-Adh and Ka-Adh-dup are not significantly different. However, Ks-Adh values are significantly lower than Ks-Adh-dup, which are more variable. In agreement with other reports, lower Ks values forAdh correlate with a high level of gene expression and relatively high percentage of G+C content in the third codon position, while the opposite applies toAdh-dup. b) At the protein level, amino acid comparisons reveal conserved regions shared by ADH and ADH-DUP, which have been assigned to known functional domains. Key residues for dehydrogenasic function are also found in ADH-DUP, thus pointing to a dehydrogenase activity for ADH-DUP, albeit very different from that of ADH.     

T14484C and T14502C in the mitochondrial ND6 gene are associated with Leber's hereditary optic neuropathy in a Chinese family

Leber's hereditary optic neuropathy (LHON) is a maternally inherited ocular disease which has been associated with three primary mitochondrial DNA mutations: G3640A, G11778A, and T14484C. In this study, we clinically characterized a Chinese family with complete penetrance of LHON. The patients in the family presented with variable clinical features. By direct DNA sequence analysis, we identified both T14484C mutation and a nearby T to C variant at nucleotide 14502 of mitochondria DNA. The T14502C variant altered I58 to V of the protein ND6, which was present in all patients of the family, but not in four unaffected family members and 200 normal controls. The co-existence of both T14484C mutation and T14502C substitution in all patients from the same LHON family suggests that T14502C may play a synergistic role with the primary mutation T14484C. The two variants together may account for the complete penetrance and absence of marked gender bias and visual recovery in the Chinese LHON family although we cannot exclude the possibility of simultaneous involvement of additional mitochondrial variant(s).     

Complete mitochondrial genome of Tubulipora flabellaris (Bryozoa: Stenolaemata): the first representative from the class Stenolaemata with unique gene order

Mitochondrial genomes play a significant role in the reconstruction of phylogenetic relationships within metazoans. There are still many controversies concerning the phylogenetic position of the phylum Bryozoa. In this research, we have finished the complete mitochondrial genome of one bryozoan (Tubulipora flabellaris), which is the first representative from the class Stenolaemata. The complete mitochondrial genome of T. flabellaris is 13,763 bp in length and contains 36 genes, which lacks the atp8 gene in contrast to the typical metazoan mitochondrial genomes. Gene arrangement comparisons indicate that the mitochondrial genome of T. flabellaris has unique gene order when compared with other metazoans. The four known bryozoans complete mitochondrial genomes also have very different gene arrangements, indicates that bryozoan mitochondrial genomes have experienced drastic rearrangements. To investigate the phylogenetic relationship of Bryozoa, phylogenetic analyses based on amino acid sequences of 11 protein coding genes (excluding atp6 and atp8) from 26 metazoan complete mitochondrial genomes were made utilizing Maximum Likelihood (ML) and Bayesian methods, respectively. The results indicate the monopoly of Lophotrochozoa and a close relationship between Chaetognatha and Bryozoa. However, more evidences are needed to clarify the relationship between two groups. Lophophorate appeared to be polyphyletic according to our analyses. Meanwhile, neither analysis supports close relationship between Branchiopod and Phoronida. Four bryozoans form a clade and the relationship among them is T. flabellaris + (F. hispida + (B. neritina + W. subtorquata)), which is in coincidence with traditional classification system.