COPPER AMINE OXIDASE1 （CuAO1） of Arabidopsis thaliana Contributes to Abscisic Acid- and Polyamine-lnduced Nitric Oxide Biosynthesis and Abscisic Acid Signal Transduction

Polyamines （PA）, polyamine oxidases, copper amine oxidases, and nitric oxide （NO） play important roles in physiology and stress responses in plants. NO biosynthesis as a result of catabolism of PA by polyamine oxidases and copper amine oxidases may explain in part PA-mediated responses. Involvement of a copper amine oxidase gene, COPPER AMINE OXIDASEI （CuAO1）, of Arabidopsis was tested for its role in stress responses using the knockouts cuao1-1 and cuaol-2. PA-induced and ABA-induced NO production investigated bY fluorometry and fluorescence microscopy showed that the cuaol-1 and cuaol-2 are impaired in NO production, suggesting a function of CuAO1 in PA and ABA-mediated NO production. Furthermore, we found a PA-dependent increase in protein S-nitrosylation. The addition of PA and ABA also resulted in H2O2 increases, cuao1-1 and cuao1-2 showed less sensitivity to exogenous ABA supplementation during germination, seedling establishment, and root growth inhibition as compared to wild-type. In response to ABA treatment, expression levels of the stress-responsive genes RD29A and ADH1 were significantly lower in the knockouts. These observations characterize cuao1-1 and cuao1-2 as ABA-insensitive mutants. Taken together, our findings extend the ABA signal transduction network to include CuAO1 as one potential contributor to enhanced NO production by ABA.

Benthodytes occidentpalauta sp.nov.,a new species of deep-sea holothuroid(Elasipodida:Psychropotidae)from the west of Kyushu-Palau Ridge in the Western Pacific Ocean

Benthodytes occidentpalauta sp.nov.was collected from the Kyushu-Palau Ridge at a depth of 5481 m in 2021.This new species is characterized by a gelatinous body wall,violet skin,six pairs of dorsal papillae,and a rough mid-ventral surface without tube feet.The dorsal deposits are rod-shaped and tripartite.Two types of papillae deposits as crosses with four arms with central bipartite apophyses.Ventral deposits are rods.Tentacle ossicles are rod-shaped with end protrusions.Gonad deposits are rodshaped,tripartite,and cross-shaped.The phylogenetic analyses based on cytochrome oxidase subunit 1(COI)and 16S individually and a concatenated dataset of COI and 16S genes of this species support that B.occidentpalauta sp.nov.belongs to Benthodytes.

Multiplex gene editing reduces oxalate production in primary hyperoxaluria type 1

Targeting key enzymes that generate oxalate precursors or substrates is an alternative strategy to eliminate primary hyperoxaluria type I(PH1),the most common and lifethreatening type of primary hyperoxaluria.The compact Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)from the Prevotella and Francisella 1(Cpf1)protein simplifies multiplex gene editing and allows for all-in-one adeno-associated virus(AAV)delivery.We hypothesized that the multiplex capabilities of the Cpf1system could help minimize oxalate formation in PH1 by simultaneously targeting the hepatic hydroxyacid oxidase 1(Hao1)and lactate dehydrogenase A(Ldha)genes.Study cohorts included treated PH1 rats(Agxt Q84X rats injected with AAV-AsCpf1 at 7 days of age),phosphate-buffered saline(PBS)-injected PH1 rats,untreated PH1 rats,and age-matched wild-type(WT)rats.The most efficient and specific CRISPR RNA(crRNA)pairs targeting the rat Hao1and Ldha genes were initially screened ex vivo.In vivo experiments demonstrated efficient genome editing of the Hao1 and Ldha genes,primarily resulting in small deletions.This resulted in decreased transcription and translational expression of Hao1 and Ldha.Treatment significantly reduced urine oxalate levels,reduced kidney damage,and alleviated nephrocalcinosis in rats with PH1.No liver toxicity,ex-liver genome editing,or obvious offtarget effects were detected.We demonstrated the AAVAsCpf1 system can target multiple genes and rescue the pathogenic phenotype in PH1,serving as a proof-ofconcept for the development of multiplex genome editingbased gene therapy.

美洲钩虫及十二指肠钩虫细胞色素C氧化酶1基因测序

目的：比较美洲钩虫及十二指肠钩虫线粒体细胞色素Ｃ氧化酶１（ＣＯ１）基因序列，确定两种钩虫间ＣＯ１基因差异。方法：现场采集钩虫成虫样本，蛋白酶Ｋ消化抽提线粒体基因组ＤＮＡ，ＰＣＲ方法扩增ＣＯ１基因，ＤＮＡ测序及分析。结果：ＰＣＲ扩增获得约７００ｂｐ美洲钩虫及十二指肠钩虫ＣＯ１基因片段，其序列比较显示两种钩虫ＣＯ１基因同源性达８９．７％，存在特定的核苷酸差异。结论：ＣＯ１基因可作为鉴别两种人体钩虫虫种标记性基因。

日本血吸虫 NADH脱氢酶 1和细胞色素 C氧化酶 1基因部分序列(英文)

目的 :测定和比较日本血吸虫浙江株和安徽株 NADH脱氢酶 1( NADH1)和细胞色素 C氧化酶 1( Cytc1)基因部分序列。方法 :用 PCR方法从特定的引物扩增出 NADH1和 Cytc1DNA基因 ,然后进行测序。结果 :获得两株日本血吸虫成虫 NADH1和 Cytc1基因及其序列。结论 :两株日本血吸虫成虫 NADH1基因及其部分序列除第 382位核苷酸不同 ( C/T)外 ,其余的均相同。两株日本血吸虫成虫 Cytc1基因及其部分序列完全相同。

Partial Sequence Analysis of Mitochondrial COI Gene of the Chinese Shrimp,Fenneropenaeus Chinensis

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235bp(28.0%), 307bp(36.6%), 138bp(16.5%) and 158bp(18.9%), respectively. Furthermore, 556bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups:Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.

Zooplankton community analysis in the Changjiang River estuary by single-gene-targeted metagenomics

DNA barcoding provides accurate stages. Single-gene-targeted metagenomic analysis identification of zooplankton species through all life based on DNA barcode databases can facilitate long- term monitoring of zooplankton communities. With the help of the available zooplankton databases, the zooplankton community of the Changjiang （Yangtze） River estuary was studied using a single-gene-targeted metagenomic method to estimate the species richness of this community. A total of 856 mitocbondrial cytochrome oxidase subunit 1 （coxl） gene sequences were determined. The environmental barcodes were clustered into 70 molecular operational taxonomic units （MOTUs）. Forty-two MOTUs matched barcoded marine organisms with more than 90% similarity and were assigned to either the species （similarity〉96%） or genus level （similarity〈96%）. Sibling species could also be distinguished. Many species that were overlooked by morphological methods were identified by molecular methods, especially gelatinous zooplankton and merozooplankton that were likely sampled at different life history phases. Zooplankton community structures differed significantly among all of the samples. The MOTU spatial distributions were influenced by the ecological habits of the corresponding species. In conclusion, single-gene-targeted metagenomic analysis is a useful tool for zooplankton studies, with which specimens from all life history stages can be identified quickly and effectively with a comprehensive database.

A new record of Sebastes koreanus from China based on morphological characters and DNA barcoding

A new record of Sebastes koreanus （Kim and Lee, 1994） was documented based on morphological characters and DNA barcoding. Fifty-six Sebastes specimens were collected from the coastal waters of northern China. Samples were identified as S. koreanus based on morphological characters. The coloration and morphometric measurements were consistent with those described from specimens collected in South Korea. In this study, specimens had the following morphological characteristics： light brown body with dark stripes and tiny dark spots, 4-5 wide indistinct vertical patterns on the side, 2 radial stripes behind and below the eyes, 1 large dark blotch on the opercle. Additionally, the following meristie characters were recorded： dorsal fin XIV-13, pectoral fin 16, anal fin III-6-7, pelvic fin [-5, lateral line scales 29-30, and vertebrae 26. The fragment of cytochrome oxidase subunit I （CO1） gene of mitochondrial DNA was sequenced for phylogenetic analysis. The mean genetic distance within the species was 0.3%. Net genetic distances between S. koreanus and other Sebastes species ranged from 3.1% to 7.6%, which was greater than the threshold for species delimitation. The phylogenetic analysis strongly supports the validity of S. koreanus in China at the genetic level. The origion, evolution, patterns of speciation and unique features in genome divergence among primate lineages of this species still need future directions of research.

Prevalence and genotype distribution of Enterobius vermicularis among kindergarteners in Shiraz and Khorramabad cities, Iran

Objective: To study the prevalence and genotype of Enterobius(E.)vermicularis from adhesive tape samples in the cities of Shiraz and Khorramabad, Iran. Methods: A total of 1 000 adhesive tape samples from kindergartens in Shiraz(500 samples) and Khorramabad(500 samples) were collected and tested using a microscope to find E. vermicularis egg/s. A questionnaire was filled out for each sample. In order to characterize the genotype of E. vermicularis, the PCR-sequencing method of the mitochondrial cytochrome C oxidase subunit 1(cox1) gene was used. Genomic DNA was extracted from the positive scotch tape samples of E. vermicularis. The cox1 gene was amplified by the polymerase chain reaction and sequenced. The sequence data were aligned using the BioEdit software and compared with the published sequences in GenBank. Phylogenetic analysis was performed using the maximum likelihood method. Results: The parasitological method showed that 15 out of the 500 samples from Shiraz(3.00%) and 12 out of the 500 samples from Khorramabad(2.40%) were infected with E. vermicularis eggs. BLAST analysis indicated that the sequenced isolates belonged to E. vermicularis genotype B while three different haplotypes were also identified. Conclusions: This is the first study on genotyping E. vermicularis in the cities of Shiraz and Khorramabad. Considering the public health importance of the disease, further studies are necessary to characterize the genotype of E. vermicularis in human populations from other regions of Iran.

Genetic variation and phylogenetic relationship of Hypoderaeum conoideum(Bloch, 1782) Dietz, 1909(Trematoda: Echinostomatidae) inferred from nuclear and mitochondrial DNA sequences

Objective:To explore genetic variations of Hypoderaeum conoideum collected from domestic ducks from 12 different localities in Thailand and Lao PDR,as well as their phylogenetic relationship with American and European isolates.Methods:The nucleotide sequences of their nuclear ribosomal DNA(ITS),mitochondrial cytochrome c oxidase subunit 1(CO1),and NADH dehydrogenase subunit 1(ND1)were used to analyze genetic diversity indices.Results:We found relatively high levels of nucleotide polymorphism in ND1(4.02%),whereas moderate and low levels were observed in CO1(2.11%)and ITS(0.96%),respectively.Based on these polymorphisms,the 20 ND1,12 CO1,and 18 ITS haplotypes were classified,and several common haplotypes were observed in all samples.At least three major lineages,namely American,European and Asian lineages,have been classified by phylogenetic analyses based on ND1 sequences.Conclusions:Our report demonstrates that the ND1 gene is the most suitable genetic marker to explore genetic variation and phylogenetic relationship of Hypoderaeum conoideum.However,a combination of all loci for ND1,CO1 and ITS would be of great value toward further genetic investigation of this endemic worldwide parasite.Thus,comprehensive molecular genetic analyses of Hypoderaeum conoideum from its worldwide distribution is needed to further understanding of the evolutionary and systematic relationships of this parasite.

Cryptic species composition and genetic diversity within Bemisia tabaci complex in soybean in India revealed by mtCOI DNA sequence

Bemisia tabaci is a cryptic species complex, causing signiifcant loss on many agricultural y important crops worldwide. Knowledge on species composition and diversity within B. tabaci complex is critical for evolving sustainable pest management strategies. Here we investigate the whitelfy species complex in soybean in major soybean growing states of India. The mitochondrial cytochrome oxidase gene subunit-1 (mtCOI) based phylogenetic relationships established using Bayesian methods indicated the existence of three cryptic species namely Asia I, Asia II 1, and Asia II 7. Al the haplotypes detected in the study could be assigned to these three cryptic species fol owing the species demarcation criteria of 3.5%divergence threshold. Of these, Asia II 1 was found to be predominant with wide spread distribution across the surveyed regions from cool temperate zones to hot and humid tropical plains. On the contrary, cryptic species Asia II 7 showed localized distribu-tion. The Asia II 1 exhibited the highest haplotype diversity and Asia I showed high level of nucleotide diversity. There was a signiifcantly high genetic differentiation among these three cryptic species. The MEAM 1, a dreadful invasive species was not detected in the specimens tested in the current study. The diversity and distribution of three cryptic species is discussed in the light of current knowledge on distribution of whitelfy species in India and yel ow mosaic disease observed during sampling survey.

Microevolution of Mitochondrial Cytochrome oxidase subunit I in Drosophila melanogaster at ＂Evolution Canyon＂, Israel

We determined the sequence of mitochondrial Cytochrome oxidase subunit I （CO1） in two Drosophila melanogaster strains originating at ＂Evolution Canyon＂, Israel. CO1 nucleotide sequences from two iso-female strains, 2-1 and 6-1, were 1,534 and 1,543 base-pairs, respectively. In each strain, ATAA was used in initiation of translation. Exchange rates for nucleotide and amino acid sequences were larger in the 6-1 strain than in the 2-1 strain when Oregon-R was used as the standard. Non-synonymous exchange rate was larger than synonymous exchange rate among the three strains.

硫唑嘌呤致AOX1和MOCOS基因型药品不良反应相关性分析

目的以核苷二磷酸连接部分X型基序15(NUDT15)rs116855232作为对照位点,分析醛氧化酶1(AOX1)rs55754655和钼辅因子硫化酶(MOCOS)rs594445基因多态性对硫唑嘌呤所致药品不良反应相关性。方法招募曾经接受或正在服用硫唑嘌呤的患者,收集静脉血。采用直接测序法测定上述位点基因型,高效液相色谱法检测红细胞中硫唑嘌呤活性代谢产物6-硫鸟嘌呤核苷酸血药浓度,监测患者药品不良反应。结果 80例患者中AOX1突变杂合子(AG)2例,无纯合突变;MOCOS突变杂合子(CA)24例,纯合突变(AA)4例;NUDT15突变杂合子(CT)16例,未发现纯合突变。Logistic回归分析表明,MOCOS突变对硫唑嘌呤所致骨髓抑制有保护性(P=0.012);与硫唑嘌呤所致流感样症状和脱发相关(P=0.043,0.030);NUDT15突变与硫唑嘌呤所致骨髓抑制相关(P=0.012);其他药品不良反应与上述位点突变无相关性。6-硫鸟嘌呤核苷酸血药浓度在上述位点野生型和突变型之间差异无统计学意义(P> 0.05)。NUDT15 rs116855232与MOCOS rs594445联合分析,MOCOS野生型且NUDT15突变型患者发生骨髓抑制的风险会更高(OR=18.40, P=0.001)。结论硫唑嘌呤首次用药前检测患者MOCOS rs594445和NUDT15rs116855232基因型,可提高用药安全性。

Modulation of peroxisomes abundance by argan oil and lipopolysaccharides in acyl-CoA oxidase 1-deficient fibroblasts

Pseudo-neonatal adrenoleukodystrophy (P-NALD) is a neurodegenerative disorder caused by acyl-CoA oxidase 1 (ACOX1) deficiency with subsequent impairment of peroxisomal fatty acid β-oxidation, accumulation of very long chain fatty acids (VLCFAs) and strong reduction in peroxisome abundance. Increase in peroxisome number has been previously suggested to improve peroxisomal disorders, and in this perspective, the present work was aimed at exploring whether modulation of peroxisomes abundance could be achieved in P-NALD fibroblasts. Here we showed that treatment with the natural Argan oil induced peroxisome proliferation in P-NALD fibroblasts. This induction was independent on activations of both nuclear receptor PPARα and its coactivator PGC-1α. Lipopolysaccharides (LPS) treatment, which caused inflammation, induced also a peroxisome proliferation that, in contrast, was dependent on activations of PPARα and PGC-1α. By its ability to induce peroxisome proliferation, Argan oil is suggested to be of potential therapeutic use in patients with P-NALD.

IL-10与子痫前期氧化应激发病机制的相关性研究

目的探讨IL-10在子痫前期氧化应激发病机制中的作用。方法采集经剖宫产子痫前期患者及同期正常妊娠孕妇胎盘组织各15例为研究对象。用实时定量PCR检测胎盘组织IL-10、NOX1 mRNA的表达。比色法检测胎盘组织MDA含量。统计比较数据并相关分析。结果与正常妊娠组相比较,子痫前期患者胎盘IL-10 mRNA表达水平降低(P<0.05)。NOX1 mRNA表达水平升高(P<0.05)。差异有统计学意义。同时MDA含量明显升高(P<0.01)。相关分析显示子痫前期患者胎盘组织IL-10 mRNA表达水平与MDA含量呈负相关(P<0.05)。与NOX1 mRNA表达水平呈负相关(P<0.01)。结论子痫前期患者胎盘组织氧化应激水平升高,与IL-10密切相关。IL-10可能是通过降低NOX1的表达来减轻胎盘氧化应激。

Highly expressed Nl-acetylpolyamine oxidase detoxifies polyamine analogue N^1-cyclopropylmethyl-N^11-ethylnorspermine in human lung cancer cell line A549

Background The critical roles of polyamines in cell growth and differentiation have made polyamJne metabolic pathway a promising target for antitumor therapy. Recent studies have demonstrated in vitro that some antitumor polyamine analogues could be used as substrates and oxidized by purified recombinant human N^1-acetylpolyamine oxidase （APAO, an enzyme that catabolizes natural poiyamines）, indicating a potential role of APAO in determining the sensitivity of cancer cells to specific antitumor analogues. This study evaluated, in vivo, the effect of APAO on cytotoxicity of antitumor polyamine analogue, N^1-cyclopropylmethyl-N^11-ethylnorspermine （CPENS） and its mechanism when highly expressed in human lung cancer line A549. Methods A clone with high expression of APAO was obtained by transfecting A549 lung cancer ceil line with pcDNA3.1/APAO plasmid and selecting with quantitative realtime PCR and APAO activity assay. Cell proliferation was determined by MTT method and apoptosis related events were evaluated by DNA fragmentation, sub-G1/flow cytometric assay, western blotting （for cytochrome C and Bax） and colorimetric assay （for casapse-3 activity）. Results A clone highly expressing APAO was obtained. High expression of APAO in A549 cells inhibited accumulation of CPENS, decreased their sensitivity to the toxicity of CPENS and prevented CPENS induced apoptosis. Conclusion These results indicate a new drug resisting, mechanism in the tumor cells. High expression of APAO can greatly decrease the sensitivity of tumor cells to the specific polyamine analogues by detoxifying those analogues and prevent analogue induced apoptosis.

Effect of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi Compatibility on Uric Acid Metabolism and Urinary Neutrophil Gelatinase-Associated Lipocalin and Kidney Injury Molecule-1 in Rats with Hyperuricemia

Objective：To explore the effects of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomicompatibility（PR） on uric acid metabolism and the expression of urinary neutrophil gelatinase-associated lipocalin（NGAL） and kidney injury molecule-1（KIM-1） in rats with hyperuricemia. Methods：Seventy male Sprague Dawley（SD） rats were randomly divided into 7 groups with 10 rats per group, including the normal group, model group, allopurinol group, benzbromarone group and PR groups at 3 doses（3.5, 7, 14 g/kg）. Except the normal group, rats of the other groups were intragastrically administered 100 mg/kg hypoxanthine and 250 mg/kg ethambutol, and subcutaneously injected with 200 mg/kg potassium oxonate. All rats were continuously modeled for 17 days, and gavaged with corresponding drugs. The rats of the normal and model groups were gavaged with saline, once a day, for 2 weeks. The levels of serum uric acid（SUA）, blood urea nitrogen（BUN） and creatinine（Cr） were determined. In addition, the contents of NGAL and KIM-1 in urine and the m RNA and protein expressions of xanthine oxidase（XOD） in liver of hyperuricemia rats were measured by reverse transcription polymerase chain reaction（RT-PCR） and Western blot, respectively. Moreover, the pathological changes of kidney were analyzed by hematoxylin and eosin（HE） stain method. Results：Compared with the normal group, the levels of SUA, BUN, NGAL and KIM-1 and the expressions of hepatic XOD m RNA and protein in the hyperuricemia rats were increased significantly（P〈0.01）. PR significantly decreased the levels of SUA, BUN, NGAL and KIM-1 and down-regulated the m RNA and protein expressions of hepatic XOD（P〈0.05 or P〈0.01）. In addition, the pathological changes of kidney were significantly suppressed by oral administration of PR. Conclusions：PR ameliorated uric acid metabolism and protected renal function, the underlying mechanism was mediated by decreasing the levels of SUA, BUN, NGAL and KIM-1, inhibiting the expression of hepatic XOD and ameliorating the pathological change of kidney.