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Identification of the causative mutation in the ITGB2 gene in a LAD1 patient by whole exome sequencing 通过全外显子组测序鉴定LAD1患者ITGB2基因的致病突变
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/15886
Nguyễn Thị Kim Liên, Nguyen Thi Van Anh, Nguyen Van Tung, Duong Anh Linh, Nguyen Thi Phương Mai, Nguyen Huy Hoang
Leukocyte adhesion deficiency type 1 (LAD1) is a rare congenital immunodeficiency disease. The cause of disease is determined to be the mutations in the ITGB2 gene that codes for CD18, the beta chain of beta-2 integrins, leads to decreased expression or functioning of CD18. This deficiency leads to severe impairment of leukocyte adhesion to the vascular wall and leukocyte migration to sites of infection and inflammation. LAD1 has also been associated with inhibition of interleukin-23 and interleukin-17 resulting in a hyperinflammatory and chronic inflammation. Patients with LAD1 typically present in early infancy with recurrent, life threatening infections that are frequently fatal before 2 years of age  without hematopoietic stem cell transplant (HSCT). However, LAD1 is difficult to diagnose and many LAD1 patients die at a young age despite intensive antibiotic therapy. Accurate diagnosis requires detailed clinical information (delayed umbilical cord loss, severe periodontitis, delayed wound healing and sores, skin abscesses, and recurrent infection), and confirmation the absence of integrins by flow cytometric analysis. A better understanding of the molecular characteristics of this disease is needed to raise awareness and definitive diagnosis infants with LAD1. To definitive diagnosis, whole exome sequencing and Sanger sequencing were performed in an eighteen-month-old boy with severe leukocytosis, recurrent infections, delayed wound healing, and hepatosplenomegaly associated with an acquired cytomegalovirus infection. Two variants: One previously reported mutation (c.533C>T, p.Pro178Leu) and one novel variant (c.59-1G>A), in the ITGB2 gene were detected. These results can be used for definitive genetic diagnosis, genetic counseling, as well as a prenatal diagnosis in LAD1 patients.
白细胞粘附缺陷1型(LAD1)是一种罕见的先天性免疫缺陷疾病。该疾病的病因被确定为编码CD18 (β -2整合素的β链)的ITGB2基因突变,导致CD18的表达或功能下降。这种缺陷导致白细胞粘附血管壁和白细胞迁移到感染和炎症部位的严重损害。LAD1还与白介素-23和白介素-17的抑制有关,导致高炎症和慢性炎症。LAD1患者通常出现在婴儿期早期,并伴有复发性、危及生命的感染,通常在2岁前死亡,不进行造血干细胞移植(HSCT)。然而,LAD1很难诊断,尽管进行了密集的抗生素治疗,许多LAD1患者仍在年轻时死亡。准确的诊断需要详细的临床资料(延迟性脐带丢失、严重牙周炎、伤口愈合和溃疡延迟、皮肤脓肿和复发性感染),并通过流式细胞分析确认整合素的缺失。需要更好地了解这种疾病的分子特征,以提高对LAD1婴儿的认识和明确诊断。为了明确诊断,我们对一个18个月大的男孩进行了全外显子组测序和Sanger测序,他患有严重的白细胞增多症,反复感染,伤口愈合延迟,肝脾肿大并伴有获得性巨细胞病毒感染。在ITGB2基因中检测到两个变体:一个先前报道的突变(c.533C>T, p.Pro178Leu)和一个新的变体(c.59-1G>A)。这些结果可用于明确的遗传诊断,遗传咨询,以及产前诊断的LAD1患者。
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引用次数: 1
A study on biological activities of Bouea macrophylla Griff leaf extract 大叶菊叶提取物的生物活性研究
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/16059
P. Hai Ha, Tran Kien Cuong, Nguyen Thi Thu Nha, Nguyen Phan Hoang Lan, N. H. Thuan Anh, Bach Long Giang, Pham Minh Quan, Tran Quoc Toan, Pham Quoc Long, Nguyen Thanh Luan
Bouea macrophylla in the family Anacardiaceae is a prominent fruit in Southeast Asia, especially in Vietnam with several prospects. This study focused on the bioactive components and biological effects of B. macrophylla ethanol leaf extract. Phytochemical analysis revealed various compounds including polyphenol, tannin, saponin, reducing agent, flavonoid, and essential oil in the extract. The DPPH assay showed the remarkable antioxidant capacity of the extract (IC50 = 6.4 ± 0.08 μg/ml), which is stronger than ascorbic acid (IC50 = 9.35 ± 0.12 μg/ml). The disc diffusion method and minimal inhibitory concentration test displayed the significant antibacterial ability of the extract against Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, and Bacillus cereus at the range from 10 - 15 mg/ml, but no activity against Staphylococcus aureus. Moreover, the MTT assay was performed the noticeable anticancer activity of the extract in HepG2 (IC50 = 72.33 ± 0.68 μg/ml), and MCF-7 cells (IC50 = 50.25 ± 1.36 μg/ml). Taken together, this study indicated the effects of B. macrophylla ethanol leaf extract from Vinh Long Province, Vietnam on antibacterial, antioxidant, and anticancer properties. Therefore, B. macrophylla possibly is used in the food industry and medicine.
大叶莲是一种重要的东南亚水果,特别是在越南,具有广阔的发展前景。本文主要研究了巨叶白杨乙醇叶提取物的生物活性成分及其生物效应。植物化学分析显示,提取物中含有多酚、单宁、皂苷、还原剂、类黄酮和精油等多种化合物。DPPH实验显示,提取物具有显著的抗氧化能力(IC50 = 6.4±0.08 μg/ml),强于抗坏血酸(IC50 = 9.35±0.12 μg/ml)。盘扩散法和最小抑菌浓度试验表明,提取物在10 ~ 15 mg/ml范围内对单核增生李斯特菌、大肠杆菌、铜绿假单胞菌、鼠伤寒沙门菌和蜡样芽孢杆菌具有显著的抑菌能力,但对金黄色葡萄球菌无抑菌活性。MTT实验显示,提取物对HepG2细胞(IC50 = 72.33±0.68 μg/ml)和MCF-7细胞(IC50 = 50.25±1.36 μg/ml)具有明显的抗肿瘤活性。综上所述,本研究表明了越南永隆省巨叶白杨乙醇叶提取物的抗菌、抗氧化和抗癌作用。因此,大叶双歧杆菌有可能用于食品工业和医药。
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引用次数: 0
In silico study of cytochrome P450 alleles and phenotypic distribution in Vietnamese population 越南人群细胞色素P450等位基因的基因分析及表型分布
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/16265
Pham Ngoc Ha, Nguyen Anh Tuan, Trinh Thi Xuan, Truong Nam Hai, Tran Dang Hung, N. Cuong
Cytochrome P450 enzymes play an important role in phase I drug metabolism, accounting for approximately 75% of the enzymatic processes. We investigated the allele and phenotypic distributions of five important CYP genes (CYP2B6, CYP3A5, CYP2C9, CYP2C19, CYP2D6) in the Vietnamese population by using Stargazer and the PharmCAT tool to call star alleles and translating them into phenotypes based on the available dataset of PharmGKB. We compared our computational analysis of the Vietnamese distributions with those of East Asia, Europe, America and other super populations, as well as with previous experimental research. The allele frequencies and phenotypic distributions of the five important CYP genes in the Vietnamese population are similar to those in East Asia while significantly different from other populations. In silico analysis also provided consistent results with previous experimental studies. In addition, the resultant data from our research contributes to the database of genetic variations in pharmacogenetics and constructs the fundamentals for future basic and applied research. 
细胞色素P450酶在I期药物代谢中起重要作用,约占酶促过程的75%。本研究利用Stargazer和PharmCAT软件对越南人群中5个CYP基因(CYP2B6、CYP3A5、CYP2C9、CYP2C19、CYP2D6)进行明星等位基因的调用,并基于phargkb数据库将其翻译成表型,研究了越南人群中CYP2B6、CYP3A5、cyp2a9、CYP2D6的等位基因和表型分布。我们将越南种群分布的计算分析与东亚、欧洲、美洲和其他超级种群的分布进行了比较,并与以往的实验研究进行了比较。越南人群中5个重要CYP基因的等位基因频率和表型分布与东亚人群相似,但与其他人群有显著差异。计算机分析也提供了与以往实验研究一致的结果。此外,我们的研究结果有助于建立药物遗传学遗传变异数据库,为今后的基础研究和应用研究奠定基础。
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引用次数: 0
Purification and characterization of recombinant nattokinase from Bacillus subtilis R0H1 枯草芽孢杆菌R0H1重组纳豆激酶的纯化及特性研究
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/16027
Nguyen Thi Thuy Ngan, Le Tuan, N. Huong
Nattokinase (NK) is a fibrinolytic enzyme with the potential for fighting cardiovascular diseases (CVD) thanks to its antithrombotic, antihypertensive, anticoagulant, anti-atherosclerotic, and neuroprotective effects. Nattokinase was first discovered and purified from soybean fermented food by Bacillus subtilis natto. To enhance NK’s activity and simplify downstream processes, production of recombinant NK using several microbial expression systems such as Escherichia coli, B. subtilis, and Lactococcus lactic has been studied. Among all of them, B. subtilis is a prominent host for overproduction of functional proteins which can be secreted directly into the culture medium. In this study, recombinant NK from B. subtilis R0H1 was purified using two-step membrane filtration. Results showed 3.2-fold increase in activity and a recovery rate of more than 80%. Molecular weight of NK was approximately 28 kDa and its fibrinolytic degradation capacity was proved according to SDS-PAGE. The optimal pH and temperature of this NK were 8.5 and 55°C, respectively. The enzyme activity was boosted by Mg2+, Ca2+ and obviously inhibited by Co2+, Zn+2, Fe2+, and SDS. The apparent Km and Vmax with fibrin as the substrate were 3.08 mM and 6.7 nmol/min, respectively. The results suggested that membrane filtration is a useful method for purification of recombinant NK from B. subtilis R0H1. Therefore, application of membrane system is proposed to purify NK at the pilot scale. In addition, our findings indicated that recombinant NK produced in B. subtilis R0H1 showed high and stable proteolytic activity in slightly alkaline pH and at high temperature. It also exhibited strong fibrinolytic activity again both substrates: fibrinogen and fibrin.
纳豆激酶(NK)是一种纤维蛋白溶解酶,具有抗血栓、抗高血压、抗凝、抗动脉粥样硬化和神经保护作用,具有对抗心血管疾病(CVD)的潜力。纳豆激酶是由纳豆枯草芽孢杆菌首次从大豆发酵食品中发现并纯化的。为了提高NK的活性并简化下游工艺,研究人员利用大肠杆菌、枯草芽孢杆菌和乳酸乳球菌等几种微生物表达系统生产重组NK。其中枯草芽孢杆菌是大量产生功能蛋白的重要宿主,可直接分泌到培养基中。本研究采用两步膜过滤纯化枯草芽孢杆菌R0H1的重组NK。结果表明,活性提高3.2倍,回收率达80%以上。NK分子量约为28 kDa, SDS-PAGE证实其具有纤溶降解能力。该NK的最适pH为8.5℃,最适温度为55℃。Mg2+、Ca2+对酶活性有促进作用,Co2+、Zn+、Fe2+和SDS对酶活性有明显抑制作用。以纤维蛋白为底物的表观Km和Vmax分别为3.08 mM和6.7 nmol/min。结果表明,膜过滤是纯化枯草芽孢杆菌R0H1重组NK的有效方法。因此,建议在中试规模上应用膜系统纯化NK。此外,我们的研究结果表明,在枯草芽孢杆菌R0H1中产生的重组NK在微碱性和高温下表现出高而稳定的蛋白水解活性。它还表现出很强的纤维蛋白溶解活性,无论是底物:纤维蛋白原还是纤维蛋白。
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引用次数: 0
Optimization of Citrus limon peel extraction, determination of main chemical components and effectiveness in repellency against Aedes mosquito dengue fever vector 柑橘柠檬皮提取工艺优化、主要化学成分及对登革热媒介伊蚊的驱避效果测定
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/16365
Phung Thi Kim Hue, Tran Van Loc, Le Tri Vien, Le Dung Sy, Ho Viet Hieu, L. Nhung, Le Thanh Do, Pham Thi Minh Khoa
Currently, scientific publications are focusing on the pharmacological actions of Citrus limon's extract and essential oil. Its essential oil is rich in bioactive monoterpenoids, such as D-limonene, β-pinene, γ-terpinene. Citrus limon essential oils have been recognized as the potential source of natural insecticides because of their selectivity, ability to be biologically degraded into non-toxic compounds, low impacts on biodiversity and the environment. Many previous studies have reported anti-bacterial, anti-fungal, anti-inflammatory, anti-cancer, hepato-regenerative, and cardio-protective activities of Citrus limon essential oil. In this study, we collected the discarded Citrus limon peel source in Gia Lai province to utilize as a material to build an optimized extraction process with the following criteria: extraction solvent, solvent/sample ratio, extraction temperature, and extraction time. The results showed a stable extraction process with a maximum extraction efficiency of 4.02%, at 40°C, for 3 hours, with two extraction times using 95% ethanol for solvent. Using GC/MS method, the determined limonene content accounted for 12.2% of the extract. The Citrus limon peel extract exhibited potency against Aedes aegypti (arbovirus vector) at a concentration of 0.01 mL, with protection time of 70 minutes and biting percentage of 0.9%, compared to negative control with statistically significant (P < 0.05). The above results correspond with the most recent publications about the effects of mosquito repellence of certain plant-based essential oils. This study has proven that Citrus limon peel in this locality signifies a promising candidature for future studies regarding its main active compound, limonene, in the control of dengue-transmitting vectors. Therefore, Citrus limon peel extract brings hope to develop new mosquito repellency products in the future.
目前,科学出版物的重点是柑橘柠檬提取物和精油的药理作用。其精油富含d -柠檬烯、β-蒎烯、γ-萜烯等生物活性单萜类物质。柑橘柠檬精油因其具有选择性、可生物降解为无毒化合物、对生物多样性和环境影响小等特点,已被公认为天然杀虫剂的潜在来源。许多研究报道了柠檬精油的抗菌、抗真菌、抗炎、抗癌、肝再生和心脏保护作用。本研究以嘉莱省废弃柠檬皮为原料,以提取溶剂、溶剂/样品比、提取温度、提取时间为标准,建立了最佳提取工艺。结果表明,以95%乙醇为溶剂,在40°C条件下提取3 h,提取2次,提取率为4.02%。GC/MS法测定的柠檬烯含量占提取液的12.2%。柠檬皮提取物在浓度为0.01 mL时,对埃及伊蚊(虫媒病毒)的防护时间为70 min,叮咬率为0.9%,与阴性对照比较,差异有统计学意义(P < 0.05)。上述结果与最近发表的关于某些植物性精油驱蚊效果的文章一致。本研究证明,该地区柠檬皮的主要活性化合物柠檬烯在控制登革热传播媒介方面具有潜在的研究价值。因此,柠檬皮提取物为今后开发新型驱蚊产品带来了希望。
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引用次数: 1
Analysis of sequence polymorphism of OsHKT1;5 gene in rice (Oryza sativa L.) 水稻OsHKT1;5基因序列多态性分析
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/15969
Nguyen Huy Duong, Tran Xuan An, Do Thi Phuc
In salt stress conditions, the excessive accumulation of Na+ ion in the cytoplasm could cause ion toxicity, metabolic imbalances, and reduction in growth of plant. The ability of maintaining a low Na+ concentrations or low Na+/K+ ratio in cytosol plays the important roles of plant adapt to salinity stress. In rice, the HKT transporter family has been proven to be critical for salt tolerance and the maintenance of Na+, K+ homeostasis under salinity conditions. In this study, we conducted the sequence polymorphisms analysis in the OsHKT1;5 coding region in order to detect the potential allelic variants in some local rice cultivars in Viet Nam. The exon 1 region and exon 2-3 region of OsHKT1;5 gene were amplified by PCR in two separate reactions. After direct sequencing of PCR products, the full length coding region of OsHKT1;5 gene was obtained and compared to reference Nipponbare OsHKT1;5 coding sequence.  Accordingly, twelve nucleotide substitutions in coding sequence of OsHKT1;5 were identified, including six non-synonymous substitutions (G382A, C418G, G551A, C994G, C1183G, A1630C) and the other six synonymous substitutions (G225A, C1038G, G1152A, G1261A, G1304C, C1608T).  Further analysis revealed that six non-synonymous substitutions (G382A, C418G, G551A, C994G, C1183G, A1630C) caused the changes in amino acids (D128N, P140A, R184H, H332D, V395L, and S544R). The single nucleotide polymorphism A1630C leading substitution of S544R was newly found in the Chiem Rong cultivar. In silico analysis of protein structure and post-translational modifications indicated that amino acid substitutions had no influence on protein structure but led to changes in post-translational modifications such as phosphorylation and SUMOylation.
在盐胁迫条件下,细胞质中Na+离子的过量积累会引起离子中毒、代谢失衡和植物生长减慢。细胞质中维持低Na+浓度或低Na+/K+比的能力对植物适应盐胁迫起着重要作用。在水稻中,HKT转运蛋白家族已被证明在盐胁迫条件下对耐盐性和维持Na+、K+稳态至关重要。在本研究中,我们对OsHKT1;5编码区进行了序列多态性分析,以检测越南一些地方水稻品种的潜在等位基因变异。OsHKT1;5基因外显子1区和外显子2-3区分别通过PCR扩增。对PCR产物直接测序,得到OsHKT1;5基因的全长编码区,并与参考Nipponbare OsHKT1;5编码序列进行比较。据此,在OsHKT1编码序列中鉴定出12个核苷酸替换,其中非同义替换6个(G382A、C418G、G551A、C994G、C1183G、A1630C),同义替换6个(G225A、C1038G、G1152A、G1261A、G1304C、C1608T)。进一步分析发现,6个非同义替换(G382A、C418G、G551A、C994G、C1183G、A1630C)引起了氨基酸(D128N、P140A、R184H、H332D、V395L和S544R)的变化。在锦戎品种中新发现了以S544R为先导的单核苷酸多态性A1630C。蛋白质结构和翻译后修饰的硅分析表明,氨基酸取代对蛋白质结构没有影响,但会导致磷酸化和SUMOylation等翻译后修饰的变化。
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引用次数: 0
The first record of Metacordyceps neogunnii (Metacordyceps, Clavicipitaceae) isolated from larva of Lepidoptera in Vietnam: morphological, phylogenetic characterization and chemical constituent analysis 越南鳞翅目新奥古尼元虫草(Metacordyceps neogunnii)的首次分离记录:形态、系统发育特征及化学成分分析
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/16033
L. D. Thuan, L. Thuy, Le Cong Hac, Nguyễn Hoàng Mai, N. Giảng, Truong Binh Nguyen
An entomopathogenic fungus, specimen DL0091 parasitized on the larvae of Lepidoptera, was collected from Lang Biang Biosphere Reserve, located in Lam Dong Province, Vietnam. The specimen DL0091 has been analyzed to be contained numerous chemical constituents, especially containing adenosine of 634 mg/Kg and cordycepin of 35.2 mg/Kg. Due to containing many bioactive compounds, DL0091 was promised to be a precious natural source that could be applied in fields of medicine and function food for health care. For classification, based on the morphology analysis, it was identified as Metacordyceps neogunnii (Metacordyceps, Clavicipitaceae) sharing the similar characteristics of M. neogunnii T.C. Wen & K.D. Hyde. Morphology of this species differed from Cordyceps neogunnii (Berk.) Berk., by many characteristics, such as the larger stroma of DL0091 (15–130 mm x 2–6 mm), of asci (550–680 μm × 5–8 μm), etc. Additionally, the combined multi-gene phylogenetic analysis, including ITS, Tef and Rpb1, well supported its systematic position in the clade of M. neogunnii, which was used as traditional herb in China and other Asian countries. In summary, DL0091 was identified as M. neogunnii, containing many bioactive compounds, could be used as the medicinal potential in human healthcare.
摘要在越南林同省朗比昂生物圈保护区采集到一种寄生在鳞翅目幼虫上的昆虫病原真菌DL0091。经分析,样品DL0091含有多种化学成分,其中腺苷含量为634 mg/Kg,虫草素含量为35.2 mg/Kg。由于DL0091含有多种生物活性化合物,因此有望成为一种宝贵的天然资源,可用于医药和保健功能食品领域。分类方面,根据形态分析,鉴定为Metacordyceps neogunnii (Metacordyceps, Clavicipitaceae),与m.n ogunnii T.C. Wen & K.D. Hyde具有相似的特征。本种形态与新冬虫夏草(Berk.)不同。傻瓜。DL0091具有较大的基质(15-130 mm × 2-6 mm)、asci (550-680 μm × 5-8 μm)等特点。此外,ITS、Tef和Rpb1等多基因的系统发育分析也支持了其在中国和其他亚洲国家作为传统草药的新乌根菌分支中的系统地位。综上所述,DL0091被鉴定为新乌氏芽孢杆菌,含有多种生物活性化合物,在人类医疗保健中具有很大的药用潜力。
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引用次数: 0
Isolation and molecular identification of obligate thermophiles from hot springs in Ba Ria – Vung Tau and Khanh Hoa provinces, Vietnam 越南巴里亚省和庆化省温泉中专性嗜热菌的分离和分子鉴定
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/15863
Tran Kha Mong, Le Thi Thanh Van, Nguyen Vu Phong, Nguyen Huu Tri
Environments with temperatures from 50°C to 80°C are rare in nature and are almost exclusively associated with geothermal regions including hot springs, solar-heated soils and volcanic areas. Thermophilic bacteria already exist and prefer in such habitats. Since innate tolerance to thermal environment and potential chassis for extracellular enzymes such as lipase, protease and amylase, which are utilized widely in the industrial fermentation, thermophilic bacteria have been becoming one of the objects for microbiologists worldwide, recently. This study aimed to isolate and identify thermophilic bacteria from hot springs in several provinces in Vietnam such as Ba Ria - Vung Tau and Khanh Hoa. In the results, six moderate thermophilic bacterial strains (namely BM7, BS5, NS1, NS3, NS4, and NW6) that could grow at 55oC were purified from the hot spring ecosystems. All micro morphology of isolates were recorded as rod-shaped, Gram positive, and endospore forming. The results of 16S rDNA sequencing and phylogenetic analysis showed that these isolate belonged to group I of Bacillus genus (the thermophilic group). The isolated strains NS1, NS3, NS4, BS5, NW6 and BM7 were identified to belong to the Bacillus genus, species as Bacillus sp. Resulting strains are potential candidates for industrial applications due to its stable fitness in a hash environment, particularly at high temperature. In addition, this study provides a useful insight into the diverse community of thermophilic bacteria (Bacillus spp.) in several hot springs of Vietnam, that can be applied as bacterial cell factories to produce biomaterials, biofuels, or valuable compounds in the future.
温度在50°C到80°C之间的环境在自然界中是罕见的,几乎完全与地热地区有关,包括温泉、太阳能加热的土壤和火山地区。嗜热细菌已经存在,并且更喜欢这样的栖息地。由于嗜热细菌对热环境的固有耐受性和在工业发酵中广泛应用的细胞外酶如脂肪酶、蛋白酶和淀粉酶的潜在基础,近年来成为世界微生物学家研究的对象之一。本研究旨在分离和鉴定越南几个省的温泉中的嗜热细菌,如巴里亚湾-青头和庆化。结果从温泉生态系统中分离到6株能在55℃条件下生长的中等嗜热细菌(BM7、BS5、NS1、NS3、NS4和NW6)。分离菌株的微形态均为杆状、革兰氏阳性和内孢子形成。16S rDNA测序和系统发育分析结果表明,该菌株属于芽孢杆菌属I群(嗜热菌群)。经鉴定,分离得到的菌株NS1、NS3、NS4、BS5、NW6和BM7属于芽孢杆菌属,属芽孢杆菌属。该菌株在高温环境下具有稳定的适应性,具有潜在的工业应用价值。此外,该研究为越南几个温泉中的嗜热细菌(芽孢杆菌)的多样化群落提供了有用的见解,这些细菌可以作为细菌细胞工厂在未来生产生物材料,生物燃料或有价值的化合物。
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引用次数: 0
Insight into Paphiopedilum x Dalatense Aver. (Orchidaceae) origin based on morphological and molecular markers 对罂粟属植物的认识。基于形态学和分子标记的兰科植物起源
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/15909
Tran Thai Vinh, H’ Yon Nie Bing, Vu Kim Cong, D. Tham, Ngo Phuong Linh, Le Ngoc Trieu, Nông Văn Duy
Paphiopedilum callosum, Paphiopedilum villosum and Paphiopedilum ´ dalatense are endangered species and are currently listed as Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES, 2019) Appendix I species. Thus, the international trade for these naturally occurring species is forbidden. However, the in vitro propagated materials and their inter-species hybrids forced by breeders can be traded for commercial purposes. P. ´ dalatense is an endemic slipper orchid species of Vietnam with high aesthetic value. Currently, the natural reserves of this species have been greatly reduced by overexploitation. In this study, P. ´ dalatense was demonstrated to be the reversible hybrid species between P. callosum and P. villosum based on both morphological and molecular markers. P. ´ dalatense possesses intermediate morphological characteristics between P. callosum and P. villosum which could be recorded in leaf, in flower and its components such as lip, petal, and dorsal sepal. There were 23 sites in ITS1-5.8S rRNA-ITS2 region sequence of P. ´ dalatense which possess the nucleotide characteristic of both P. callosum and P. villosum. There were two out of three P. ´ dalatense investigated samples possessed partial matK gene which was identical to the corresponding sequence in P. villosum and the last sample possessed identical partial matK gene to P. callosum. Proving P. ´ dalatense to be the reversible hybrid species between P. callosum and P. villosum is the essentially scientific base for commercial breeding of P. ´ dalatense by artificial hybridization which contributes to avoid overexploiting the natural P. ´ dalatense resources.
花心罂粟、绒毛罂粟和dalatense Paphiopedilum是濒危物种,目前被列为《濒危野生动植物种国际贸易公约》(CITES, 2019)附录I物种。因此,这些天然物种的国际贸易是被禁止的。然而,由育种者强迫的离体繁殖材料及其种间杂交种可以用于商业目的进行交易。dalatense是越南特有的一种拖鞋兰,具有很高的审美价值。目前,该物种的自然保护区因过度开发而大大减少。本研究通过形态学和分子标记证明了dalatense是胼胝体和毛囊体的可逆杂交物种。dalatense具有介于胼胝体和绒毛体之间的中间形态特征,包括叶片、花及其组成部分(唇、花瓣和背萼片)。dalatense的ITS1-5.8S rRNA-ITS2区序列中有23个位点同时具有胼胝体和绒毛体的核苷酸特征。3个dalatense样品中有2个样品的部分matK基因序列与绒毛草相同,最后一个样品的部分matK基因序列与胼胝体相同。证明大理石竹是胼胝体与毛茛的可逆杂交品种,是大理石竹人工杂交育种的重要科学依据,有助于避免大理石竹天然资源的过度开发。
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引用次数: 0
Total phenolic, flavonoid contents and antioxidant activity of tamarind seed and pulp extracts 罗望子籽和果肉提取物的总酚、类黄酮含量及抗氧化活性
Pub Date : 2022-06-30 DOI: 10.15625/1811-4989/15930
Le Phuong Ha, Nguyen Van Ngoc, Nguyen Thi Trang Huyen, Le Thi Thu Hang, Nguyen Thi Kieu Oanh, Tran Mai Thi Tuyet, Nguyen Thi Mai Phuong, N. M. Minh Hong
Tamarind (Tamarindus indica) has long been known for its high nutrition content and pharmacological potential. However, there is lack of studies on the content of antioxidants, phenolic and flavonoid contents of tamarind seed grown in Vietnam. Thus, the aim of this study was to compare the seeds and pulps of Tamarindus indica from three different areas across Vietnam including Son La, Hai Phong and Sai Gon with regard to the total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity of their water and methanol extracts, as well as their cytotoxicity on a normal BKH-21cells. TPC and TFC were evaluated by the Folin–Ciocalteu reagent and aluminum chloride, respectively. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging assays were used to investigate antioxidant capacity. The safety of T. indica extracts was assessed by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Our results showed that the methanolic extracts yielded higher TPC (742.919 ± 50.360 mg GAE/g extract), TFC (68.492 ± 0.023 mg QE/g extract) and possessed stronger free radical scavenging activity (IC50 of 52.5 µg/mL) compared to that of water extracts. T. indica seeds from all three regions possessed higher TPC, TFC and antioxidant activity than those of pulps. Regarding the safety, in vitro analysis showed that tamarind seed and pulp extracts only became toxic to BHK-21 cell line at a very high concentration with IC50 values range from 143.77 µg/mL to 620.35 µg/mL. This study revealed that T. indica seeds and pulps can serve as functional food as well as potential antioxidants in pharmaceutical products.
罗望子(Tamarindus indica)长期以来以其高营养含量和药理潜力而闻名。然而,对越南产罗望子中抗氧化剂、酚类和类黄酮含量的研究较少。因此,本研究的目的是比较越南三个不同地区(山腊、海防和西贡)的柽柳种子和果肉的总酚含量(TPC)、总黄酮含量(TFC)、水提取物和甲醇提取物的抗氧化活性,以及它们对正常bhk -21细胞的细胞毒性。分别用Folin-Ciocalteu试剂和氯化铝评价TPC和TFC。采用2,2-二苯基-1-吡啶肼(DPPH)和2,2'-氮化萘(3-乙基苯并噻唑-6-磺酸(ABTS))自由基清除实验研究抗氧化能力。采用MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑)测定法评价籼稻提取物的安全性。结果表明,与水提取物相比,甲醇提取物的TPC(742.919±50.360 mg GAE/g提取物)和TFC(68.492±0.023 mg QE/g提取物)具有更高的自由基清除活性(IC50为52.5µg/mL)。三个地区的籼稻种子的TPC、TFC和抗氧化活性均高于纸浆。体外安全性分析表明,罗望子籽和果肉提取物仅在非常高浓度时才对BHK-21细胞株产生毒性,IC50值为143.77 ~ 620.35µg/mL。本研究表明,籼稻种子和果肉既可作为功能性食品,又可作为潜在的抗氧化剂用于医药产品。
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引用次数: 1
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Vietnam Journal of Biotechnology
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