病毒性脑膜脑炎262例血清线粒体天冬氨酸转氨酶同工酶变化的临床意义

目的　探讨病毒性脑膜脑炎 (病脑 )患儿血清线粒体天冬氨酸转氨酶同工酶 (m AST)变化及其临床意义。方法　应用全自动生化分析仪测定 2 62例病脑患儿和 12 1例健康儿童静脉血清中m AST、天冬氨酸转氨酶总活性 (t AST)活性 ,并计算m AST/t AST。结果　轻度病脑组血清m AST活性 15.50± 3 .91U/L ,中度组3 4 .79± 7.2 5U/L ,重度组 55.76± 11.3 3U/L ,正常对照组 7.93± 2 .80U/L ,各组间比较差异有显著性 (P均 <0 .0 5) ;m AST/t AST值 ,轻度患儿组 (3 6.76± 8.51) % ,中度患儿组 (47.63± 11.53 ) % ,重度患儿组 (61.81±7.3 3 ) % ,正常对照组 (2 5.2 6± 9.2 1) % ,各组间比较差异亦有显著性 (P均 <0 .0 5)。结论　测定血清m AST活性及m AST/t AST可作为判断小儿病脑病情及预后的指标之一     

Mitochondrial DNA Copy Number Is Associated With Treatment Response and Cognitive Function in Euthymic Bipolar Patients Receiving Valproate

BackgroundBipolar disorder (BD) is associated with cognitive impairment and mitochondrial dysfunction. However, the associations among mitochondrial DNA copy number (MCN), treatment response, and cognitive function remain elusive in BD patients.MethodsSixty euthymic BD patients receiving valproate (VPA) and 66 healthy controls from the community were recruited. The indices of metabolic syndrome (MetS) were measured. Quantitative polymerase chain reaction analysis of blood leukocytes was used to measure the MCN. Cognitive function was measured by calculating perseverative errors and completed categories on the Wisconsin Card Sorting Test (WCST). The VPA treatment response was measured using the Alda scale.ResultsBD patients had significantly higher MCN, triglyceride, and C-reactive protein (CRP) levels, waist circumference, and worse performance on the WCST than the controls. Regression models showed that BD itself and the VPA concentration exerted significant effects on increased MCN levels. Moreover, the receiver operating characteristic curve analysis showed that an MCN of 2.05 distinguished VPA responders from nonresponders, with an area under the curve of 0.705 and a sensitivity and specificity of 0.529 and 0.816, respectively. An MCN level ≥2.05 was associated with 5.39 higher odds of being a VPA responder (P = .006). BD patients who were stratified into the high-MCN group had a higher VPA response rate, better WCST performance, lower CRP level, and less MetS.ConclusionsThe study suggests a link between the peripheral MCN and cognitive function in BD patients. As an inflammatory status, MetS might modulate this association.     

Oxaliplatin-loaded nanoemulsion containing Teucrium polium L. essential oil induces apoptosis in Colon cancer cell lines through ROS-mediated pathway

Oxaliplatin (Oxa)-associated adverse side effects have considerably limited the clinical use of the drug in colon cancer therapy. Mutant p53 has diverse mutational profiles in colon cancer, and it influences the potencies of various chemotherapeutic drugs, including Oxa. Thus, it would be highly beneficial to identify an alternative therapeutic strategy that not only reduces the toxicity of Oxa, but also exerts a synergistic effect against colon cancers, regardless of their p53 profiles. The present study was aimed at preparing and optimizing Teucrium polium L. essential oil nanoemulsion (TPO-NANO) and investigating its effect on the sensitivity of colon cancer cells with differences in p53 status (HCT116 wild-type and HT-29 mutant-type) to Oxa. The viability of treated cells was determined and the combination index (CI) was calculated. Morphological changes were determined under inverted microscopy, while percentage apoptosis was assayed using flow cytometry. Intracellular ROS and the protein levels of p53 and Bax were measured. The colony-forming potential of treated cells was determined using colony assay. The size of TPO-NANO was markedly increased from 12.90 ± 0.04 nm to 14.47 ± 0.53 nm after loading Oxa (p ≤ 0.05). The combination (Oxa + TPO-NANO) produced a synergetic effect in HCT116 and HT-29, with CI of 0.94 and 0.88, respectively. Microscopic examination and flow cytometric analysis revealed that cells treated with Oxa + TPO-NANO had a higher percentage of apoptosis than cells exposed to monotherapy. Cumulatively, Oxa exerted an apoptotic effect on wild or mutant p53 colon cancer cells when combined with TPO-NANO, through a mechanism involving ROS-mediated mitochondrial apoptosis.     

Corosolic acid analogue,a natural triterpenoid saponin,induces apoptosis on human hepatocarcinoma cells through mitochondrial pathway in vitro

Context 2a,-3a,-24-Trihydroxyurs-12-en-28-oic acid (TEO, a corosolic acid analogue) is a triterpenoid saponin isolated from Actinidia valvata Dunn (Actinidiaceae), a well-known traditional Chinese medicine.

Objective This study investigated the anti-proliferation and inducing apoptosis effects of TEO in three human hepatocellular carcinoma (HCC) cell lines.

Materials and methods Cytotoxic activity of TEO was determined by the MTT assay at various concentrations from 2.5 to 40?μg/mL in BEL-7402, BEL-7404 and SMMC-7721 cell lines. Cell morphology was assessed by acridine orange/ethidium bromide and 4′-6-diamidino-2-phenylindole dihydrochloride staining and fluorescence microscopy. Cell-cycle distribution and DNA damage were determined by flow cytometry and comet assay. Mitochondrial dysfunction was assessed by JC-1 staining and transmission electron microscopy. Apoptosis changes were explored by Western blot, TNF-α and caspase-3, -8, -9 assays.

Results TEO exhibited inhibition effects on BEL-7402, BEL-7404 and SMMC-7721 cells treated for 24?h, the IC₅₀ values were 34.6, 30.8 and 30.5?μg/mL, respectively. TEO (40?μg/mL)-treated three cell lines increased by more than 21% in the G1 phase and presented the morphological change and DNA damage. TEO also declined the mitochondrial membrane potential and altered mitochondrial ultra-structure. Furthermore, caspase-3, caspase-8, caspase-9 and TNF-α were also activated. Mechanism investigation showed that TEO could decrease anti-apoptotic Bcl-2 protein expression, increase proapoptotic Bax and Bid proteins expressions and increase Bax/Bcl-2 ratio.

Conclusion Our results demonstrate for the first time that TEO inhibited growth of HCC cell lines and induced G1 phase arrest. Moreover, proapoptotic effects of TEO were mediated through the activation of TNF-α, caspases and mitochondrial pathway.     

乌鲁木齐地区线粒体12SrRNA基因突变筛查分析

目的　研究乌鲁木齐地区非综合征性聋患者线粒体12S rRNA基因突变情况。方法　收集乌鲁木齐非综合征性聋患者标本609例,对其进行临床和分子遗传学评估。结果　12S rRNA基因突变分析共发现11个突变位点,已知的A1555G、961DelT、C1494T突变分别占2.96%,1.15%,0.16%。另外A1047G突变相关报道较少,A1585G突变未见相关报道。其他突变均为多态性位点。结论　线粒体12S rRNA突变是引起遗传性聋的重要因素,此次乌鲁木齐地区线粒体12S rRNA A1555G突变在聋病人群中的检出率与前期报道相比有所降低,可能与耳毒性药物使用量降低有关。A1047G与新发现的A1585G突变是否与聋相关,还需进一步研究。对筛查中阳性突变携带者及其母系家庭成员需告知氨基糖苷类抗生素使用风险,使其避免使用,逐步降低药物性聋的发生率。     

Single nucleotide polymorphisms in the D-loop region of mitochondrial DNA and age-at-onset of patients with chronic kidney disease

Background The mitochondrial displacement loop （D-loop） accumulates mutations and single nucleotide polymorphisms （SNPs） at a higher frequency than other regions of mitochondrial DNA （mtDNA）.We previously identified disease riskassociated SNPs in the D-loop of chronic kidney disease （CKD） patients; in this study,we investigated the association of age-at-onset and D-loop SNPs in CKD patients.Methods The D-loop region of mtDNA was sequenced in 119 CKD patients attending the Fourth Hospital of Hebei Medical University between 2002 and 2008.The age-at-onset curve of the CKD patients was calculated using the KaplanMeier method at each SNP site,and compared using the log-rank test.Results The mean age of 119 CKD patients was （55.6±14.2） years,and 56.3% were males.The mean estimated glomerular filtration rate （eGFR） was （81.2±12.4） ml·min^-1·1.73 m^-2,with 79.8% （n=95） of patients having an eGFR 〈60 ml·min^-1·1.73 m^-2.All participants had an eGFR 〉30 ml·min^-1·1.73 m^-2.The age-at-onset for CKD patients who smoked was significantly lower than that of non-smoking CKD patients.The SNP sites of nucleotides 150C/T were identified for their association with age-at-onset using the log-rank test.The age-at-onset of patients with the minor allele T genotype was significantly lower than that of patients with the C genotype at the 150 SNP site （P=0.010）.Conclusions Genetic polymorphisms in the D-loop appear to be predictive markers for age-at-onset in CKD patients.Accordingly,the analysis of genetic polymorphisms in the mitochondrial D-loop may help identify CKD patient subgroups at high risk of early onset disease.     

艾灸对衰老模型大鼠线粒体DNA含量的影响

目的:探讨艾灸对衰老模型大鼠肝细胞线粒体DNA含量的影响。方法:将36只SD大鼠随机分为3组:青年对照组、衰老模型组、艾灸治疗组。采用25%D-半乳糖溶液125mg/kg·d-1皮下注射复制衰老大鼠模型,采用紫外分光光度法观察艾灸对衰老模型大鼠肝细胞线粒体DNA含量的影响。结果:与青年对照组比较,衰老模型组大鼠肝细胞线粒体DNA含量明显增加(P<0.05);与衰老模型组比较,艾灸治疗组大鼠肝细胞线粒体DNA含量明显减少(P<0.05)。结论:艾灸可以减少肝细胞线粒体DNA的含量,从而起到延缓衰老的作用。     

线粒体H2S供体AP39对心肌梗死大鼠心肌纤维化的影响及其与线粒体动力学的关系

目的]已有研究表明H₂S可拮抗心肌纤维化,但线粒体靶向性H₂S能否拮抗心肌梗死后心肌纤维化,且是否与调控线粒体融合与分裂有关目前并不明确。为了探究这一关系,进行了该研究。 [方法]在动物实验中予以异丙肾上腺素[ISO,50 mg/(kg·d)]腹腔注射构建SD大鼠心肌梗死模型,对各组大鼠行心电图检测,使用线粒体H₂S供体AP39[36 μg/(kg·d)],腹腔注射连续处理SD大鼠4周,使用Masson染色检测心肌纤维化情况,使用Western blot检测相关蛋白表达情况。体外实验以氯化钴(CoCl₂,800 μmol/L)诱导H9c2心肌细胞缺氧损伤,AP39(100 nmol/L)处理H9c2细胞,使用DL-炔丙基甘氨酸(PAG,2 mmol/L)抑制内源性硫化氢合成酶胱硫醚-γ-裂解酶(CSE),并通过荧光探针检测心肌细胞活性氧(ROS)的水平。 [结果]梗死大鼠心肌存在明显间质纤维化,胶原纤维大量堆积,且CSE、线粒体融合蛋白2(MFN2)表达下调,线粒体动力相关蛋白1(DRP1)表达增加,AP39干预后则可明显改善以上变化,而加入CSE抑制剂PAG则可逆转AP39的以上作用。同时在体外实验中发现,以CoCl₂诱导H9c2心肌细胞缺氧损伤时,细胞内ROS水平升高,MFN2表达下调,DRP1表达增加,AP39则可上调MFN2蛋白表达,抑制DRP1表达,降低心肌细胞ROS水平,而PAG则可逆转以上变化。 [结论]线粒体靶向性H₂S供体AP39可以改善心肌梗死大鼠心肌纤维化,且可促进线粒体融合,抑制线粒体过度分裂。     

血清线粒体型门冬氨酸转移酶在百草枯中毒大鼠的价值

目的通过检测SD大鼠血清线粒体型天门冬氨酸氨基转移酶(mAST)的浓度的变化及其余各项血气和生化指标的相关性,探讨百草枯(PQ)中毒损伤的机制及mAST在百草枯中毒早期评估多器官功能中的价值。方法将84只健康SD大鼠随机分为对照组和染毒组,灌胃后0 h、2 h、6 h、12 h、24 h、48 h及72 h分别收集动脉血做血气分析,检测大鼠mAST及其他生化指标的浓度变化。结果 mAST在PQ中毒2 h即明显升高,远远早于动脉血气及动脉血乳酸值,而与反映急性肝功能损伤的指标(AST、ALT、DBIL、TBIL)、反映肝纤维化指标(γGT)、反映肾功能损伤的指标(SRBP)及一些特异性不强的指标(CK,LDH)呈显著正相关。结论 mAST在大鼠中毒早期组织未出现缺氧时即有显著升高且与反映肝肾功能的多个生化指标正相关,提示线粒体损伤可能是百草枯中毒致多器官损伤的发病机制中独立于组织缺氧的一项重要因素,对早期评估百草枯中毒患者肝肾功能损伤程度有参考价值。