Hearing impairment-associated KARS mutations lead to defects in aminoacylation of both cytoplasmic and mitochondrial tRNALys

Aminoacyl-tRNA synthetases(aaRSs) are ubiquitously expressed, essential enzymes, synthesizing aminoacyl-tRNAs for protein synthesis. Functional defects of aaRSs frequently cause various human disorders. Human KARS encodes both cytosolic and mitochondrial lysyl-tRNA synthetases(LysRSs). Previously, two mutations(c.1129 GA and c.517 TC) were identified that led to hearing impairment; however, the underlying biochemical mechanism is unclear. In the present study, we found that the two mutations have no impact on the incorporation of LysRS into the multiple-synthetase complex in the cytosol, but affect the cytosolic LysRS level, its tertiary structure, and cytosolic tRNA aminoacylation in vitro. As for mitochondrial translation, the two mutations have little effect on the steady-state level, mitochondrial targeting, and tRNA binding affinity of mitochondrial LysRS. However, they exhibit striking differences in charging mitochondrial tRNA~(Lys), with the c.517TC mutant being completely deficient in vitro and in vivo. We constructed two yeast genetic models, which are powerful tools to test the in vivo aminoacylation activity of KARS mutations at both the cytosolic and mitochondrial levels. Overall, our data provided biochemical insights into the potentially molecular pathological mechanism of KARS c.1129GA and c.517TC mutations and provided yeast genetic bases to investigate other KARS mutations in the future.     

Anthranilic diamides derivatives as potential ryanodine receptor modulators: Synthesis,biological evaluation and structure activity relationship

A series of novel anthranilic diamides derivatives (7a–s) containing halogen, trifluoromethyl group and cyano group were designed, synthesized, and characterized by melting point, ¹H NMR, ¹³C NMR and elemental analyses. The bioactivity revealed that most of them showed moderate to excellent activities against oriental armyworm (Mythimna separata) and diamondback moth (Plutella xylostella). Above all, the larvicidal activity of 7o against oriental armyworm was 100% and 40% at 0.25 and 0.1?mg?L^?1, comparable to that of the standard chlorantraniliprole (100%, 0.25?mg?L^?1 and 20%, 0.1?mg?L^?1). What is more, 7o against diamondback moth displayed 90% insecticidal activity at 0.01?mg?L^?1, superior to chlorantraniliprole (45%, 0.01?mg?L^?1). The experiments 7o on the American cockroach (Periplaneta Americana) heart beating rates (Dorsal vessel) and contractile force were compared with chlorantraniliprole. In addition, 7o could affect the calcium homeostasis in the central neurons of the third larvae of oriental armyworm, which revealed that the ryanodine receptor is the potential target of 7o. The density functional theory (DFT) calculation results revealed the amide bridge, the benzene ring of anthraniloyl moiety and pyrazole ring might play an important role in the insecticidal activity through hydrophobic interactions and π-π conjugations.     

Diet-induced elevation of circulating HSP70 may trigger cell adhesion and promote the development of atherosclerosis in rats

Although accumulating evidence indicates that heat shock protein 70 (HSP70) could be secreted into plasma and its levels have been found to have an ambiguous association with atherosclerosis, our knowledge for the exact role of circulating HSP70 in the development of atherosclerosis is still limited. In the present study, we report an adhesion-promoting effect of exogenous HSP70 and evaluate the potential involvement of elevated circulating HSP70 in the development of atherosclerosis. Time-dependent elevation of plasma HSP70 was found in diet-induced atherosclerotic rats, whose effect was investigated through further in vitro experiments. In rat aortic endothelial cell (RAEC) cultures, exogenous HSP70 incubation neither produced cell injuries by itself nor had protective effects on cell injuries caused by Ox-LDL or homocysteine. However, exogenous HSP70 administration could lead to a higher adhesion rate between rat peripheral blood monocytes (PBMCs) and RAECs. This adhesion-promoting effect appeared only when PBMCs, rather than RAECs, were pretreated with HSP70 incubation. PBMCs in an HSP70 environment released more IL-6 to supernatant, which subsequently up-regulated the expression of ICAM-1 in RAECs. These results indicate that the diet-induced elevation of circulating HSP70 could trigger cell adhesion with the help of IL-6 as a mediator, which provides a novel possible mechanism for understanding the role of circulating HSP70 in the pathogenesis of atherosclerosis.     

Synthesis,insecticidal evaluation and mode of action of novel anthranilic diamide derivatives containing sulfur moiety as potential ryanodine receptor activators

Anthranilic diamide insecticide could control lepidopteran pests by selectively binding and activating insect ryanodine receptors (RyRs), and the unique mode of action is different from other conventional insecticides. In order to discover new anthranilic diamide insecticide as ryanodine receptors activators, a series of 11 novel anthranilic diamides derivatives (Ia-k) were synthesized and confirmed by melting point, ¹H NMR, ¹³C NMR and elemental analyses. The preliminary bioactivity revealed that most title compounds showed moderate to remarkable activities against oriental armyworm (Mythimna separata) and diamondback moth (Plutella xylostella). Especially, compounds Ia and If, which exhibited 100% larvicidal activity against oriental armyworm at 1.0?mg?L^?1, and comparable to that of chlorantraniliprole (100% at 1?mg?L^?1). If displayed 60% insecticidal activity against diamondback moth at 0.01?mg?L^?1, better than chlorantraniliprole (45% at 0.01?mg?L^?1). The preliminary structure activity relationships were discussed. In addition, the calcium imaging experiment indicated that the insect ryanodine receptor is the potential target of If.     

A lincRNA-DYNLRB2-2/GPR119/GLP-1R/ABCA1-dependent signal transduction pathway is essential for the regulation of cholesterol homeostasis

Accumulated evidence shows that G protein-coupled receptor 119 (GPR119) plays a key role in glucose and lipid metabolism. Here, we explored the effect of GPR119 on cholesterol metabolism and inflammation in THP-1 macrophages and atherosclerotic plaque progression in apoE^−/− mice. We found that oxidized LDL (Ox-LDL) significantly induced long intervening noncoding RNA (lincRNA)-DYNLRB2-2 expression, resulting in the upregulation of GPR119 and ABCA1 expression through the glucagon-like peptide 1 receptor signaling pathway. GPR119 significantly decreased cellular cholesterol content and increased apoA-I-mediated cholesterol efflux in THP-1 macrophage-derived foam cells. In vivo, apoE^−/− mice were randomly divided into two groups and infected with lentivirus (LV)-Mock or LV-GPR119 for 8 weeks. GPR119-treated mice showed decreased liver lipid content and plasma TG, interleukin (IL)-1β, IL-6, and TNF-α levels, whereas plasma levels of apoA-I were significantly increased. Consistent with this, atherosclerotic lesion development was significantly inhibited by infection of apoE^−/− mice with LV-GPR119. Our findings clearly indicate that, Ox-LDL significantly induced lincRNA-DYNLRB2-2 expression, which promoted ABCA1-mediated cholesterol efflux and inhibited inflammation through GPR119 in THP-1 macrophage-derived foam cells. Moreover, GPR119 decreased lipid and serum inflammatory cytokine levels, decreasing atherosclerosis in apoE^−/− mice. These suggest that GPR119 may be a promising candidate as a therapeutic agent.     

3种药物对老化卵母细胞4种母源mRNA水平的影响

利用Real-Time PCR技术定量分析了绵羊(Ovis aries)卵母细胞在生长-成熟-老化过程4种母源mRNA(Mater、Zar1、Dnmt1、GDF9)发生的动态变化,以及3种药物(咖啡因、DTT和矾酸盐)对老化卵母细胞母源mRNA水平的影响。结果显示,除Dnmt1在12 h开始下降外,卵母细胞中其他3个基因的表达量都是从生发泡期逐渐下降,且在体外培养至24 h降到最低,然后随着卵母细胞的老化又逐渐上升。其中GDF9和Mater 2个基因在30 h组的卵母细胞中的表达量显著高于24 h组卵母细胞(P0.05)。30 h卵母细胞中Zar1和Dnmt1的转录水平与24 h卵母细胞差异不显著(P0.05)。除Zar1外,咖啡因和DTT都显著降低了老化卵母细胞中其他3个基因的表达量(P0.05),且所有基因表达量与24 h组卵母细胞差异不显著(P0.05)。矾酸盐处理后的卵母细胞其4种母源mRNA水平都是最高,除Zar1外其他3个基因表达量都显著高于24 h组(P0.05)。结论是,Mater、Zar1、Dnmt1、GDF9 4种母源mRNA水平与卵母细胞质量成反比,咖啡因和DTT能在一定程度上延缓卵母细胞的衰老,改善卵母细胞质量;而矾酸盐则加速了卵母细胞老化进程,降低了卵母细胞质量。     

Hemoglobinopathy: Molecular Epidemiological Characteristics and Health Effects on Hakka People in the Meizhou Region,Southern China

Background

Hemoglobinopathies are the most common inherited diseases in southern China. However, there have been only a few epidemiological studies of hemoglobinopathies in Guangdong province.

Materials and Methods

Peripheral blood samples were collected from 15299 “healthy” unrelated subjects of dominantly ethnic Hakka in the Meizhou region, on which hemoglobin electrophoresis and routine blood tests were performed. Suspected cases with hemoglobin variants and hereditary persistence of fetal hemoglobin (HPFH) were further characterized by PCR, DNA sequencing, reverse dot blot (RDB) or multiplex ligation-dependent probe amplification (MLPA). In addition, 1743 samples were randomly selected from the 15299 subjects for thalassemia screening, and suspected thalassemia carriers were identified by PCR and RDB.

Results

The gene frequency of hemoglobin variants was 0.477% (73/15299). The five main subgroups of the ten hemoglobin variants were Hb E, Hb G-Chinese, Hb Q-Tahiland, Hb New York and Hb J-Bangkok. 277 cases (15.89%, 277/1743) of suspected thalassemia carriers with microcytosis (MCV<82 fl) were found by thalassemia screening, and were tested by a RDB gene chip to reveal a total of 196 mutant chromosomes: including 124 α-thalassemia mutant chromosomes and 72 β-thalassemia mutant chromosomes. These results give a heterozygote frequency of 11.24% for common α and β thalassemia in the Hakka population in the Meizhou region. 3 cases of HPFH/δβ-thalassemia were found, including 2 cases of Vietnamese HPFH (FPFH-7) and a rare Belgian ^Gγ(^Aγδβ)⁰–thalassemia identified in Chinese.

Conclusions

Our results provide a detailed prevalence and molecular characterization of hemoglobinopathies in Hakka people of the Meizhou region. The estimated numbers of pregnancies each year in the Meizhou region, in which the fetus would be at risk for β thalassemia major or intermedia, Bart’s hydrops fetalis, and Hb H disease, are 25 (95% CI, 15 to 38), 40 (95% CI, 26 to 57), and 15 (95% CI, 8 to 23), respectively.     

Mineral Elements in Root of Wild Saposhnikovia divaricata and Its Rhizosphere Soil

Mineral elements are important components of medicinal herbs, and their concentrations are affected by many factors. In this study, Ca, Mg, Na, K, Fe, Mn, Cu, and Zn concentrations in wild Saposhnikovia divaricata and its rhizosphere soil collected from seven locations at two different times in China were measured, and influences of rhizosphere soil on those minerals in plant were evaluated. The results showed that mean concentrations of eight minerals in plant samples decreased in the order: Ca > Mg > Na > K > Fe > Zn > Mn > Cu, and those in the soil samples followed the following order: Na > Fe > Ca > K > Mg > Mn > Zn > Cu. Mean concentrations of Ca, Na, Mg, and K in plants were higher than those in soils, while higher mean concentrations of the other four minerals were found in soils. It was found that there was a positive correlation of Mg, Na, and Cu concentrations in the plant with those in the soil respectively, but a negative correlation of Mn concentration in plant with that in the soil. Except Ca, K, and Mn, the other five minerals in plant were all directly affected by one or more chemical compositions of soil. The results also indicate that pH value and concentrations of total nitrogen, Mg, Mn, and Cu in soil had significant correlations with multimineral elements in plant. In a word, mineral elements uptake of S. divaricata can be changed by adjusting the soil fertility levels to meet the need of appropriate quality control of S. divaricata.