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Fangchinoline protects hepatic ischemia/reperfusion liver injury in rats through anti-oxidative stress and anti-inflammation properties: an in silico study. 方棘霉素通过抗氧化应激和抗炎特性保护大鼠肝缺血/再灌注肝损伤:一项硅学研究。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-10 DOI: 10.1002/bab.2628
Shuangxi Li, AnDong Xiang, Feng Guo, Abdullah A Alarfaj, Zehai Gao

Liver ischemia-reperfusion (I/R) injury is a common cause of organ failure, developed by a sudden block in the blood and oxygen supply and subsequent restoration. I/R damage is responsible for acute and chronic rejection after organ transplantation, accounting for 10% of early graft failure. The study investigated the therapeutic properties of fangchinoline in liver injury-induced rats. The rats were divided into three groups: Sham, I/R without pretreatment, and I/R + 10 mg/kg fangchinoline pretreatment. Blood and liver samples were collected for assays, and an in silico docking analysis was conducted to determine fangchinoline's inhibitory effect. The pretreatment with 10 mg/kg of fangchinoline effectively reduced hepatic marker enzymes such as AST, LDH, and ALT in the serum of rats with liver I/R damage. Fangchinoline treatment significantly reduced interleukin-8 (IL-8), IL-6, and tumor necrosis factor-α (TNF-α) in I/R-induced rats, boosting antioxidants and decreasing MDA. Histopathological studies showed liver injury protection, and fangchinoline inhibited TNF-α and IL-6 with improved binding affinity. Fangchinoline has hepatoprotective properties by reducing inflammation in rats with liver I/R damage, as demonstrated in the current study. Hence, it can be an effective salutary agent in preventing liver damage caused by I/R.

肝脏缺血再灌注(I/R)损伤是器官衰竭的常见原因,它是由血液和氧气供应突然中断以及随后的恢复造成的。I/R损伤是器官移植后急性和慢性排斥反应的原因,占早期移植失败的10%。本研究探讨了吩喹啉对肝损伤大鼠的治疗作用。大鼠分为三组:Sham 组、未经预处理的 I/R 组和 I/R + 10 mg/kg fangchinoline 预处理组。采集血液和肝脏样本进行化验,并进行硅学对接分析以确定芒果胆碱的抑制作用。用 10 毫克/千克的方棘霉素预处理可有效降低肝脏 I/R 损伤大鼠血清中的 AST、LDH 和 ALT 等肝脏标志酶。方棘霉素能明显降低 I/R 大鼠体内的白细胞介素-8(IL-8)、IL-6 和肿瘤坏死因子-α(TNF-α),提高抗氧化能力,降低 MDA。组织病理学研究显示,方棘霉素对肝损伤有保护作用,并能抑制 TNF-α 和 IL-6,且结合亲和力更强。本研究表明,方棘霉素具有保肝作用,能减轻肝脏 I/R 损伤大鼠的炎症反应。因此,方棘霉素可以有效地预防 I/R 引起的肝损伤。
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引用次数: 0
CircMYO1C silencing alleviates chondrocytes inflammation and apoptosis through m6A/HMGB1 axis in osteoarthritis. 沉默CircMYO1C可通过m6A/HMGB1轴缓解骨关节炎中软骨细胞的炎症和凋亡。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-09 DOI: 10.1002/bab.2635
Haitao Sun, Xudong Chu, Weiqing Qian, Hong Yin

Circular RNAs (circRNAs) are involved in osteoarthritis (OA) progression. This study aimed to investigate the role and molecular mechanisms of circMYO1C in OA. CircMYO1C was upregulated in OA- and interleukin-1β (IL-1β)-exposed chondrocytes. The results indicated that circMYO1C knockdown repressed the inflammatory factors (tumor necrosis factor alpha [TNF-α], interleukin-6 [IL-6], interleukin-8 [IL-8], etc.) and apoptosis of chondrocytes following IL-1β exposure. CircMYO1C was an N6-methyladenosine (m6A)-modified circRNA with m6A characteristics. High mobility group box 1 (HMGB1) was a target of circMYO1C. IL-1β exposure increased the stability and half-life (t1/2) of HMGB1 mRNA, while silencing circMYO1C reduced HMGB1 mRNA stability. Taken together, circMYO1C targets the m6A/HMGB1 axis to promote chondrocyte apoptosis and inflammation. The present study demonstrates that the circMYO1C/m6A/HMGB1 axis is essential for OA progression, highlighting a novel potential therapeutic target for clinical OA.

环状 RNA(circRNA)参与骨关节炎(OA)的进展。本研究旨在探讨circMYO1C在OA中的作用和分子机制。在OA和白细胞介素-1β(IL-1β)暴露的软骨细胞中,circMYO1C被上调。结果表明,circMYO1C敲除抑制了IL-1β暴露后的炎症因子(肿瘤坏死因子α [TNF-α]、白细胞介素-6 [IL-6]、白细胞介素-8 [IL-8]等)和软骨细胞的凋亡。CircMYO1C 是一种具有 m6A 特征的 N6-甲基腺苷(m6A)修饰的 circRNA。高迁移率基团框 1(HMGB1)是 circMYO1C 的靶标。IL-1β 暴露增加了 HMGB1 mRNA 的稳定性和半衰期(t1/2),而沉默 circMYO1C 则降低了 HMGB1 mRNA 的稳定性。综上所述,circMYO1C 以 m6A/HMGB1 轴为靶标,促进软骨细胞凋亡和炎症。本研究表明,circMYO1C/m6A/HMGB1 轴对 OA 的进展至关重要,为临床治疗 OA 提供了一个新的潜在治疗靶点。
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引用次数: 0
Heparosan biosynthesis in recombinant Bacillus megaterium: Influence of N-acetylglucosamine supplementation and kinetic modeling. 重组巨型芽孢杆菌中的eparosan生物合成:N-乙酰葡糖胺补充和动力学模型的影响。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-08 DOI: 10.1002/bab.2634
Ganesh Nehru, Rengesh Balakrishnan, Nivedhitha Swaminathan, Subbi Rami Reddy Tadi, Senthilkumar Sivaprakasam

Heparosan, an unsulfated polysaccharide, plays a pivotal role as a primary precursor in the biosynthesis of heparin-an influential anticoagulant with diverse therapeutic applications. To enhance heparosan production, the utilization of metabolic engineering in nonpathogenic microbial strains is emerging as a secure and promising strategy. In the investigation of heparosan production by recombinant Bacillus megaterium, a kinetic modeling approach was employed to explore the impact of initial substrate concentration and the supplementation of precursor sugars. The adapted logistic model was utilized to thoroughly analyze three vital parameters: the B. megaterium growth dynamics, sucrose utilization, and heparosan formation. It was noted that at an initial sucrose concentration of 30 g L-1 (S1), it caused an inhibitory effect on both cell growth and substrate utilization. Intriguingly, the inclusion of N-acetylglucosamine (S2) resulted in a significant 1.6-fold enhancement in heparosan concentration. In addressing the complexities of the dual substrate system involving S1 and S2, a multi-substrate kinetic models, specifically the double Andrew's model was employed. This approach not only delved into the intricacies of dual substrate kinetics but also effectively described the relationships among the primary state variables. Consequently, these models not only provide a nuanced understanding of the system's behavior but also serve as a roadmap for optimizing the design and management of the heparosan production method.

肝聚糖是一种未硫酸化的多糖,在肝素的生物合成过程中作为主要前体发挥着关键作用,肝素是一种有影响力的抗凝剂,具有多种治疗用途。为了提高肝糖的产量,在非致病性微生物菌株中利用代谢工程正在成为一种安全而有前景的策略。在研究重组巨大芽孢杆菌生产肝聚糖的过程中,采用了动力学建模方法来探讨初始底物浓度和前体糖补充的影响。利用改编的逻辑模型深入分析了三个重要参数:巨芽孢杆菌的生长动力学、蔗糖利用率和肝聚糖的形成。结果发现,当初始蔗糖浓度为 30 g L-1 时(S1),蔗糖对细胞生长和底物利用都有抑制作用。有趣的是,加入 N-乙酰葡糖胺(S2)后,肝聚糖的浓度显著提高了 1.6 倍。为了解决 S1 和 S2 双底物系统的复杂性,我们采用了多底物动力学模型,特别是双安德鲁模型。这种方法不仅深入研究了双底物动力学的复杂性,而且有效地描述了主要状态变量之间的关系。因此,这些模型不仅提供了对系统行为的细微理解,还为优化肝磷脂生产方法的设计和管理提供了路线图。
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引用次数: 0
Recent development on nanomaterial-based biosensors for identifying thyroid tumor biomarkers. 用于识别甲状腺肿瘤生物标记物的纳米材料生物传感器的最新发展。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-03 DOI: 10.1002/bab.2632
Kun Xu, XiaoLu Wu

The incidence of thyroid tumors has been increasing yearly over the past decade, making it the fourth highest tumor in women. This places various biological burdens on those affected. Currently, thyroid tumors are primarily diagnosed using percutaneous fine needle aspiration and ultrasound. However, these methods are complex, expensive, and less accurate, and they may fail to detect some thyroid nodules. As an alternative, researchers are focusing on blood-based biomarkers in addition to the traditional diagnostic methods, assisted predominantly by nanomaterials. Early identification of thyroid cancer is crucial as it is highly treatable. Various sensing systems have been developed using nanomaterial-mediated approaches to enhance the detection system. Nanomaterials are effectively applied in biosensors for surface functionalization and are conjugated with biomolecules to improve the interaction with the target analyte. This review discusses nanomaterial-assisted thyroid tumor detection, with a special focus on nanomaterial-based biosensors.

近十年来,甲状腺肿瘤的发病率逐年上升,已成为女性第四大高发肿瘤。这给患者带来了各种生理负担。目前,甲状腺肿瘤的诊断主要采用经皮细针穿刺术和超声波检查。然而,这些方法复杂、昂贵、准确性较低,而且可能无法检测到某些甲状腺结节。作为一种替代方法,除了传统的诊断方法外,研究人员还将重点放在以血液为基础的生物标志物上,主要是通过纳米材料来辅助诊断。由于甲状腺癌极易治疗,因此早期识别甲状腺癌至关重要。目前已开发出各种以纳米材料为媒介的传感系统,以增强检测系统。纳米材料可有效地应用于生物传感器的表面功能化,并与生物分子共轭,以改善与目标分析物的相互作用。本综述讨论了纳米材料辅助甲状腺肿瘤检测,特别关注基于纳米材料的生物传感器。
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引用次数: 0
Fish scale-derived hydroxyapatite for alveolar ridge preservation. 用于牙槽嵴保存的鱼鳞衍生羟基磷灰石
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-01 DOI: 10.1002/bab.2627
P M Prathibha, N G Thomas, Y B Dalvi, K G Varghese, P K Binsi, A A Zynudheen, M Lekshmi, J Shilpa, Vellappally Sajith, Anil Sukumaran

Alveolar ridge resorption following tooth extraction poses significant challenges for future dental restorations. This study investigated the efficacy of fish scale-derived hydroxyapatite (FSHA) as a socket preservation graft material to maintain alveolar bone volume and architecture. FSHA was extracted from *Labeo rohita* fish scales and characterized using Fourier transform infrared (FTIR) analysis. In vitro, biocompatibility and osteogenic potential were assessed using Saos-2 human osteosarcoma cells. Cell viability, migration, and proliferation were evaluated using MTT and scratch assays. In vivo performance was assessed in a rat model, and FSHA was compared to a commercial xenograft (Osseograft) and ungrafted controls. Histological analysis was performed at 8-week post-implantation to quantify new bone formation. FTIR confirmed the purity and homogeneity of FSHA. In vitro, FSHA enhanced Saos-2 viability, migration, and proliferation compared to controls. In vivo, FSHA demonstrated superior bone regeneration compared to Osseograft and ungrafted sites, with balanced graft resorption and new bone formation. Histological analysis revealed an active incorporation of FSHA into new bone, with minimal gaps and ongoing remodeling. Approximately 50%-60% of FSHA was resorbed by 8 weeks, closely matching the rate of new bone deposition. FSHA stimulated more bone formation in the apical socket region than in coronal areas. In conclusion, FSHA is a promising biomaterial for alveolar ridge preservation, exhibiting excellent biocompatibility, osteogenic potential, and balanced resorption. Its ability to promote robust bone regeneration highlights its potential as an effective alternative to currently used graft materials in socket preservation procedures.

拔牙后牙槽骨嵴吸收给未来的牙科修复带来了巨大挑战。本研究调查了鱼鳞衍生羟基磷灰石(FSHA)作为牙槽窝保存移植材料在保持牙槽骨体积和结构方面的功效。FSHA是从*Labeo rohita*鱼鳞中提取的,并使用傅立叶变换红外(FTIR)分析法对其进行了表征。在体外,使用 Saos-2 人骨肉瘤细胞对其生物相容性和成骨潜力进行了评估。使用 MTT 和划痕试验评估了细胞的活力、迁移和增殖。在大鼠模型中评估了 FSHA 的体内性能,并将其与商业异种移植(Osseograft)和未移植对照进行了比较。在植入后 8 周进行组织学分析,以量化新骨的形成。傅立叶变换红外光谱证实了 FSHA 的纯度和均匀性。在体外,与对照组相比,FSHA 提高了 Saos-2 的存活率、迁移率和增殖率。在体内,与 Osseograft 和未移植部位相比,FSHA 的骨再生能力更强,移植骨的吸收和新骨的形成达到了平衡。组织学分析表明,FSHA 积极融入新骨,间隙极小,并在不断重塑。大约 50%-60%的 FSHA 在 8 周前被吸收,与新骨沉积的速度非常接近。与冠状区域相比,FSHA 在牙槽骨根尖区域刺激了更多的骨形成。总之,FSHA 具有良好的生物相容性、成骨潜力和均衡的吸收能力,是一种很有前景的牙槽嵴保存生物材料。其促进骨再生的能力突显了其在牙槽窝保存手术中作为目前使用的移植材料的有效替代品的潜力。
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引用次数: 0
Screening and characterization of Chromobacterium piscinae AMA-5 for enhanced production of violacein and its ability of textile dyeing. 提高鱼腥色杆菌 AMA-5 产碱量及其纺织品染色能力的筛选和表征。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-01 DOI: 10.1002/bab.2629
Deeppal, Kishan Kumar Raj, Tapas Chowdhury, Alavala Umarajashekhar, Ravindra Soni

A violet pigment (violacein) bacterial isolate AMA-5 was isolated from soil samples collected from Achanakmar Biosphere Reserve, Mungeli district, Chhattisgarh, India. The yield of biocolor from this isolate was screened in minimal medium after 48 h of incubation at 37°C ± 2°C temperature. The violet pigment was extracted in ethanol. It was also observed that ammonium chloride (2.5 g/1000 mL) as a nitrogen source is the best to enhance AMA-5 pigment production among other nitrogen sources (ammonium sulfate, tryptophan, ammonium iron sulfate, and peptone). The Sanger sequencing of 16S rDNA of strain AMA-5 showed similarity with Chromobacterium piscinae. From the available literature and research articles, it was assumed that this violet color pigment is violacein. It was further verified by conducting high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), and proton nuclear magnetic resonance (1H-NMR) analysis. The violet biocolor that extracted was used in cotton and polyester fabric dyeing. After the fabrics treated with sodium chloride as a mordant were completely dried, it was identified that the color was solidifying. Overall study showed that C. piscinae AMA-5 has good potential for production of violacein, which is the most important industrial natural dye used to add color to textile products.

从印度恰蒂斯加尔邦 Mungeli 地区 Achanakmar 生物圈保护区的土壤样本中分离出一种紫色色素(violacein)细菌分离物 AMA-5。在 37°C ± 2°C 的温度下培养 48 小时后,在最小培养基中对该分离菌的生物色素产量进行了筛选。紫色色素用乙醇提取。还观察到,在其他氮源(硫酸铵、色氨酸、硫酸铁铵和蛋白胨)中,氯化铵(2.5 克/1000 毫升)作为氮源最能提高 AMA-5 色素的产量。菌株 AMA-5 的 16S rDNA 的桑格测序结果显示与 Chromobacterium piscinae 相似。根据现有的文献和研究文章,推测这种紫色色素为中毒性色素。通过高效液相色谱(HPLC)、傅立叶变换红外光谱(FTIR)和质子核磁共振(1H-NMR)分析进一步验证了这一点。提取的紫色生物色素可用于棉布和涤纶布的染色。用氯化钠作为媒染剂处理过的织物完全干燥后,发现颜色正在凝固。总体研究表明,C. piscinae AMA-5 具有生产紫罗兰色素的良好潜力,而紫罗兰色素是最重要的工业天然染料,可为纺织品增色。
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引用次数: 0
Effect of tannic acid on doxorubicin-induced cellular stress: Expression levels of heat shock genes in rat spleen. 单宁酸对多柔比星诱导的细胞应激的影响:大鼠脾脏中热休克基因的表达水平
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-30 DOI: 10.1002/bab.2633
Duygu Kizir, Melike Karaman, Yeliz Demir, Hamid Ceylan

Doxorubicin (DOX), an anthracycline group antibiotic, has been extensively employed as a potent chemotherapeutic agent for treating solid and hematopoietic tumors in humans. Amid exposure to diverse stress conditions, living organisms swiftly initiate the synthesis of heat shock proteins (HSPs), a set of highly conserved proteins. Tannic acid (TA) has garnered increasing study attention due to its special chemical properties, health benefits, and wide availability. This study's primary aim is to elucidate the impact of DOX and TA on the expression levels of Hsp90aa1, Hspa1a, Hspa4, and Hspa5 in the spleen tissues of rats. Sprague Dawley rats (Rattus norvegicus, male, 9-10 weeks old, 180 ± 20 g) were randomly divided into 4 groups: control, DOX (30 mg/kg cumulative), TA (50 mg/kg), and DOX + TA (5 mg/kg and 50 mg/kg, respectively). Subsequently, spleen tissues were collected from rats, and complementary DNA libraries were generated after the application process. The quantitative real-time PCR method was used to detect and quantify the mRNA expression changes of the Hsp90aa1, Hspa1a, Hspa4, and Hspa5 genes our results showed that the mRNA expressions of the targeted genes were up-regulated in rat spleen tissues exposed to DOX. However, this increase was remarkably suppressed by TA treatment. These findings suggest that TA may serve as a protective agent, mitigating the toxic effects of DOX in the rat spleen.

多柔比星(DOX)是一种蒽环类抗生素,已被广泛用作治疗人类实体瘤和造血肿瘤的强效化疗药物。在暴露于各种应激条件下时,生物体会迅速启动热休克蛋白(HSPs)的合成,这是一组高度保守的蛋白质。单宁酸(TA)因其特殊的化学性质、对健康的益处和广泛的可获得性而受到越来越多的研究关注。本研究的主要目的是阐明 DOX 和 TA 对大鼠脾脏组织中 Hsp90aa1、Hspa1a、Hspa4 和 Hspa5 表达水平的影响。将 Sprague Dawley 大鼠(Rattus norvegicus,雄性,9-10 周龄,180 ± 20 g)随机分为 4 组:对照组、DOX 组(30 mg/kg 累积)、TA 组(50 mg/kg)和 DOX + TA 组(分别为 5 mg/kg 和 50 mg/kg)。随后,收集大鼠脾脏组织,并在应用过程中生成互补 DNA 文库。结果表明,在暴露于 DOX 的大鼠脾脏组织中,目标基因的 mRNA 表达上调。然而,TA 治疗显著抑制了这种上调。这些发现表明,TA 可作为一种保护剂,减轻 DOX 对大鼠脾脏的毒性作用。
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引用次数: 0
Plasmodium falciparum cysteine protease Falcipain 3: A potential enzyme for proteolytic processing of histone acetyltransferase PfGCN5. 恶性疟原虫半胱氨酸蛋白酶 Falcipain 3:组蛋白乙酰转移酶 PfGCN5 蛋白水解加工的潜在酶。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-25 DOI: 10.1002/bab.2630
Poonam Nagar, Krishanu Bhowmick, Aishwarya Chawla, Md Zubbair Malik, Naidu Subbarao, Inderjeet Kaur, Suman Kumar Dhar

In spite of 150 years of studying malaria, the unique features of the malarial parasite, Plasmodium, still perplex researchers. One of the methods by which the parasite manages its gene expression is epigenetic regulation, the champion of which is PfGCN5, an essential enzyme responsible for acetylating histone proteins. PfGCN5 is a ∼170 kDa chromatin-remodeling enzyme that harbors the conserved bromodomain and acetyltransferase domain situated in its C-terminus domain. Although the PfGCN5 proteolytic processing is essential for its activity, the specific protease involved in this process still remains elusive. Identification of PfGCN5 interacting proteins through immunoprecipitation (IP) followed by LC-tandem mass spectrometry analysis revealed the presence of food vacuolar proteins, such as the cysteine protease Falcipain 3 (FP3), in addition to the typical members of the PfGCN5 complex. The direct interaction between FP3 and PfGCN5 was further validated by in vitro pull-down assay as well as IP assay. Subsequently, use of cysteine protease inhibitor E64d led to the inhibition of protease-specific processing of PfGCN5 with concomitant enrichment and co-localization of PfGCN5 and FP3 around the food vacuole as evidenced by confocal microscopy as well as electron microscopy. Remarkably, the proteolytic cleavage of the nuclear protein PfGCN5 by food vacuolar protease FP3 is exceptional and atypical in eukaryotic organisms. Targeting the proteolytic processing of GCN5 and the associated protease FP3 could provide a novel approach for drug development aimed at addressing the growing resistance of parasites to current antimalarial drugs.

尽管研究疟疾已有 150 年的历史,但疟原虫的独特特征仍然令研究人员感到困惑。疟原虫管理其基因表达的方法之一是表观遗传调控,其中的佼佼者是 PfGCN5,它是一种负责乙酰化组蛋白的重要酶。PfGCN5 是一种 ∼170 kDa 的染色质重塑酶,其 C 端结构域含有保守的溴结构域和乙酰转移酶结构域。虽然 PfGCN5 的蛋白水解加工对其活性至关重要,但参与这一过程的特异性蛋白酶仍然难以确定。通过免疫沉淀(IP)鉴定 PfGCN5 的相互作用蛋白,然后进行 LC-串联质谱分析,发现除了 PfGCN5 复合物的典型成员外,还存在食物空泡蛋白,如半胱氨酸蛋白酶 Falcipain 3 (FP3)。体外牵引试验和 IP 试验进一步验证了 FP3 与 PfGCN5 之间的直接相互作用。随后,使用半胱氨酸蛋白酶抑制剂 E64d 抑制了蛋白酶对 PfGCN5 的特异性加工,同时,共聚焦显微镜和电子显微镜显示,PfGCN5 和 FP3 在食物空泡周围富集和共定位。值得注意的是,在真核生物中,食物液泡蛋白酶FP3对核蛋白PfGCN5的蛋白水解是非常特殊和非典型的。以 GCN5 和相关蛋白酶 FP3 的蛋白水解加工为靶点,可以为药物开发提供一种新方法,从而解决寄生虫对当前抗疟药物的抗药性日益增强的问题。
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引用次数: 0
Regulation of hsa_circ_0112136 by m6A demethylase FTO can enhance the malignancy of gastric cancer via the regulation of the PI3K/AKT/mTOR pathway m6A去甲基化酶FTO对hsa_circ_0112136的调控可通过调控PI3K/AKT/mTOR通路增强胃癌的恶性程度
IF 2.8 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-22 DOI: 10.1002/bab.2631
Jia Liu, Xiangming Fang
A growing body of research highlights the role that N6‐methyladenosine (m6A) modification and circular RNAs (circRNAs) play in gastric cancer (GC) cases. However, studies elucidating the function and mechanism of the recently discovered circRNA hsa_circ_0112136 in GC are limited. This study aimed to examine the pathophysiology of GC progression due to fat mass and obesity‐associated protein (FTO)‐mediated N6‐methyladenosine (m6A) modification of hsa_circ_0112136. The hsa_circ_0112136 and FTO levels in the GC samples were analyzed using qRT‐PCR. The Transwell invasion assay, wound healing assay, and CCK8 assays were employed to assess alterations in GC cell invasiveness, migration, and viability due to the aberrant regulation of hsa_circ_0112136 and FTO. Phosphorylated PI3K, AKT, and mTOR (the key proteins of the PI3K/AKT/mTOR pathway) were detected via western blotting after hsa_circ_0112136 suppression. A tumor transplantation mouse model was constructed to evaluate the suppression of hsa_circ_0112136's function in vivo. The correlation among hsa_circ_0112136 and FTO was identified using the MeRIP assay.Levels of hsa_circ_0112136 and FTO were evidently elevated in GC samples. Suppression of has_circ_0112136 reduced the viability, migration, and invasive ability of GC cells in vitro, as well as delayed tumor growth in vivo via suppression of the activation of the PI3K/AKT/mTOR pathway. FTO decreased hsa_circ_0112136 m6A levels and enhanced hsa_circ_0112136 expression. Furthermore, FTO upregulation enhanced GC cell invasion, migration, and survival, which was reversed by hsa_circ_0112136 suppression.Our study proposes that hsa_circ_0112136 functions as a tumor promoter, facilitating the malignant progression of GC through m6A modification (suppressed by FTO) and activating the PI3K/AKT/mTOR pathway. This suggests that targeting FTO‐m6A‐hsa_circ_0112136‐PI3K/AKT/mTOR may be a novel approach for GC intervention.
越来越多的研究强调了 N6-甲基腺苷(m6A)修饰和环状 RNA(circRNA)在胃癌(GC)病例中的作用。然而,阐明最近发现的循环 RNA hsa_circ_0112136 在胃癌中的功能和机制的研究还很有限。本研究旨在探讨脂肪量和肥胖相关蛋白(FTO)介导的 N6-甲基腺苷(m6A)修饰 hsa_circ_0112136 导致 GC 进展的病理生理学。利用 qRT-PCR 分析了 GC 样品中的 hsa_circ_0112136 和 FTO 水平。采用 Transwell 侵袭试验、伤口愈合试验和 CCK8 试验来评估由于 hsa_circ_0112136 和 FTO 的异常调控而导致的 GC 细胞侵袭性、迁移性和活力的改变。抑制 hsa_circ_0112136 后,通过 Western 印迹法检测磷酸化的 PI3K、AKT 和 mTOR(PI3K/AKT/mTOR 通路的关键蛋白)。为了评估 hsa_circ_0112136 在体内的功能抑制作用,我们构建了一个肿瘤移植小鼠模型。利用MeRIP试验确定了hsa_circ_0112136和FTO之间的相关性。通过抑制 PI3K/AKT/mTOR 通路的激活,抑制 hsa_circ_0112136 可降低 GC 细胞在体外的存活率、迁移和侵袭能力,并延缓肿瘤在体内的生长。FTO 降低了 hsa_circ_0112136 m6A 的水平,增强了 hsa_circ_0112136 的表达。我们的研究提出,hsa_circ_0112136具有肿瘤促进剂的功能,通过m6A修饰(被FTO抑制)和激活PI3K/AKT/mTOR通路促进GC的恶性进展。这表明,靶向 FTO-m6A-hsa_circ_0112136-PI3K/AKT/mTOR 可能是干预 GC 的一种新方法。
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引用次数: 0
Identification of a novel anti‐ROR1 nanobody through phage display and its biochemical characterization 通过噬菌体展示鉴定新型抗 ROR1 纳米抗体及其生化特征
IF 2.8 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-22 DOI: 10.1002/bab.2623
Li Kang, Yingkui Dong, Wanxue Wang, Zehua Li, Yizhuo Wang, Li Yan, Cunlong Yin, XiaoHui Zhang, Han Dai, Bo Wu, Hongxin Zhao, Junfeng Wang
In this study, we aimed to develop nanobodies targeting receptor tyrosine kinase‐like orphan receptor 1 (ROR1) for cancer diagnosis and therapy. We immunized alpacas with ROR1, extracted RNA from their blood, and converted it to complementary DNA (cDNA) to amplify the VHH (variable domain of heavy‐chain antibodies) sequence. This sequence was used to construct a phage library with a capacity of 8 ×108. Screening identified a high‐affinity nanobody, HCAbs1, which binds effectively to ROR1. ELISA and surface plasmon resonance analyses revealed HCAbs1's binding affinities to ROR1 at 4.42 and 12.9 nM, respectively. Functional tests showed HCAbs1 could reduce extracellular signal‐regulated kinase (ERK) phosphorylation levels induced by Wnt5a in ROR1‐transfected cells. Our findings highlight the potential of HCAbs1 nanobodies in diagnosing and treating cancers through targeting ROR1.
在这项研究中,我们旨在开发针对受体酪氨酸激酶样孤儿受体1(ROR1)的纳米抗体,用于癌症诊断和治疗。我们用ROR1免疫羊驼,从它们的血液中提取RNA,并将其转化为互补DNA(cDNA),扩增VHH(重链抗体的可变域)序列。利用该序列构建了一个容量为 8 ×108 的噬菌体文库。筛选确定了一种高亲和力纳米抗体 HCAbs1,它能有效地与 ROR1 结合。ELISA和表面等离子共振分析显示,HCAbs1与ROR1的结合亲和力分别为4.42和12.9 nM。功能测试显示,HCAbs1能降低Wnt5a在ROR1转染细胞中诱导的细胞外信号调节激酶(ERK)磷酸化水平。我们的研究结果凸显了HCAbs1纳米抗体通过靶向ROR1诊断和治疗癌症的潜力。
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Biotechnology and applied biochemistry
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