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Development and optimization of an efficient RNA isolation protocol from bovine (Bos indicus) spermatozoa 开发和优化从牛(Bos indicus)精子中分离 RNA 的高效方案
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-02 DOI: 10.1016/j.bbrep.2024.101862
Sofi Imran Ul Umar , Sushil Prasad , Soumen Naskar , Pooja Chowdhury , Anju Rana , Pranab Jyoti Das
Achieving the optimum extraction of RNA from spermatozoal cells is crucial for carrying out effective high-throughput analysis regarding its role in fertility and other reproduction processes in Bos indicus. Nevertheless, semen comprises spermatozoa and several other secretions from the male reproductive system, which as well consist of diverse somatic cell types. Therefore, the elimination of somatic cells guarantees the purity of the sperm RNA. In the present study, we tested five different RNA isolation protocols and evaluated them for their yield and purity using spectrophotometer and polymerase chain reaction. Among the five RNA isolation protocols, the Triazol + RNAeasy plus Kit + TCEP method revealed optimum performance. We successfully achieved isolation of spermatozoal RNA without any spermatozoal DNA contamination from Bos indicus spermatozoa that contains approx. 1000 to 10,000 times less RNA as compared to other mammalian somatic cells. RNA quality was assessed using primers Protamine1 (spermatozoal RNA and spermatozoal DNA), CDH1 (epithelial cell), KIT (germ cell) and PTPRC (leukocytes) designed using primer BLAST where there was no product amplified except Prm1 whose product size was specific for spermatozoal RNA. The results of our investigation on RNA isolation procedures indicate that the inclusion of a disulphide reducing agent (TCEP) is crucial for the process of sperm cell lysis.
从精子细胞中提取最佳的 RNA 对于进行有效的高通量分析至关重要,这些分析涉及精子在豚鼠生育和其他繁殖过程中的作用。然而,精液由精子和来自雄性生殖系统的其他一些分泌物组成,其中也包括各种体细胞类型。因此,去除体细胞可保证精子 RNA 的纯度。在本研究中,我们测试了五种不同的 RNA 分离方案,并使用分光光度计和聚合酶链反应评估了它们的产量和纯度。在五种 RNA 分离方案中,Triazol + RNAeasy plus Kit + TCEP 方法的性能最佳。与其他哺乳动物体细胞相比,Bos indicus 精子中的 RNA 含量约低 1000 到 10,000 倍,我们成功地从这些精子中分离出了精子 RNA,且没有精子 DNA 污染。使用引物 BLAST 设计的引物 Protamine1(精子 RNA 和精子 DNA)、CDH1(上皮细胞)、KIT(生殖细胞)和 PTPRC(白细胞)对 RNA 质量进行了评估。我们对 RNA 分离程序的研究结果表明,加入二硫化物还原剂(TCEP)对精子细胞裂解过程至关重要。
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引用次数: 0
Identification of hub genes and pathways in Uterine corpus endometrial carcinoma (UCEC): A comprehensive in silico study 识别子宫内膜癌(UCEC)的枢纽基因和通路:一项全面的硅学研究
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 DOI: 10.1016/j.bbrep.2024.101860
Mahsa Ejlalidiz , Ameneh Mehri-Ghahfarrokhi , Mohammadreza Saberiyan

Background

Uterine corpus endometrial carcinoma (UCEC), derived from the endometrium, is the most common type of endometrial malignasis. This gynecological malignancy is very common all over the world, especially in developed countries and shows a potentially rising trend correlated with the increase in obese women.

Methods

Differentially Expressed Genes (DEGs) analysis was conducted on GSE7305 and GSE25628 datasets from the Gene Expression Omnibus (GEO). DEGs were identified using GEO2R (adjusted p-value <0.05, |logFC| > 1). Pathway analysis employed KEGG and Gene Ontology databases, while protein-protein interactions were analyzed using Cytoscape and Gephi. GEPIA was used for target gene validation.

Results

We have identified 304 common DEGs and 78 hub genes using GEO and PPI analysis, respectively. The GO and KEGG pathways analysis revealed enrichment of DEGs in extracellular matrix structural constituent, extracellular space, cell adhesion, and ECM-receptor interaction. GEPIA analysis identified three genes, ENG, GNG4, and ECT2, whose expression significantly differed between normal and tumor samples.

Conclusion

This analysis study identified the hub genes and associated pathways involved in the pathogenesis of UCEC. The identified hub genes exhibit remarkable potential as diagnostic biomarkers, providing a significant opportunity for early diagnosis and more effective therapeutic approaches for UCEC.
背景来自子宫内膜的子宫体子宫内膜癌(UCEC)是最常见的子宫内膜恶性肿瘤。这种妇科恶性肿瘤在全世界都很常见,尤其是在发达国家,而且随着肥胖妇女的增加,其发病率有可能呈上升趋势。使用 GEO2R 鉴定 DEGs(调整后 p 值为 0.05,|logFC| >1)。通路分析使用 KEGG 和基因本体数据库,蛋白质-蛋白质相互作用分析使用 Cytoscape 和 Gephi。结果我们利用 GEO 和 PPI 分析分别鉴定了 304 个常见 DEGs 和 78 个枢纽基因。GO和KEGG通路分析显示,DEGs富集于细胞外基质结构成分、细胞外基质空间、细胞粘附和ECM-受体相互作用。GEPIA分析发现了三个基因,即ENG、GNG4和ECT2,它们在正常样本和肿瘤样本中的表达有显著差异。所发现的枢纽基因具有作为诊断生物标志物的巨大潜力,为 UCEC 的早期诊断和更有效的治疗方法提供了重要机会。
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引用次数: 0
Investigation of CST7 and hsa-miR-4793-5p gene expression in breast cancer 乳腺癌中 CST7 和 hsa-miR-4793-5p 基因表达的研究
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 DOI: 10.1016/j.bbrep.2024.101863
Niloufar Sadat Kalaki , Mozhgan Ahmadzadeh , Mandana Dehghan , Venus Shahabi Rabori , Sima Davoudi , Hamed Afkhami
Breast cancer (BC) presents as a worldwide challenge, known as the most frequently diagnosed cancer in women. In 2022, BC was diagnosed in 2.3 million women with 670,000 deaths globally. In this research, our objective was to examine the CST7 and has-miR-4793-5p gene expression in BC tumor tissues and adjacent normal tissues.
Using GSE57897 gene expression data from 422 BC samples and 31 breast samples from healthy controls which was based on the Platform GPL18722 (spotted oligonucleotide Homo sapiens microRNA (miRNA) array) in the Gene Expression Omnibus (GEO) and compare with miRNAs with a conserved target location on CST7 mRNA were found using databases. The study population included 60 fresh BC tissue samples and adjacent normal tissues as control. The Quantitative Real-Time PCR was used to evaluate the expression levels of CST7 and has-miR-4793-5p in the breast tissues.
The present study, found that CST7 and hsa-miR-4793-5p were significantly increased in tumoral tissues in compare to normal tissues. Further analysis revealed a remarkable association between CST7 and hsa-miR-4793-5p gene expression alteration. ROC curve analysis demonstrated high accuracy for CST7 expression in BC tumors. Comparison of gene expression between different stages and patient family history showed significant findings. Due to the high sensitivity and specificity of the expression changes of these two genes, they are suitable candidates for further investigations to be considered as part of a diagnosis and prognosis panel.
乳腺癌(BC)是一项世界性难题,是女性最常见的癌症。2022 年,全球有 230 万女性被诊断为乳腺癌,67 万人死亡。本研究旨在检测CST7和has-miR-4793-5p基因在乳腺癌肿瘤组织和邻近正常组织中的表达。我们利用基因表达总库(Gene Expression Omnibus,GEO)中基于 GPL18722 平台(点状寡核苷酸智人 microRNA(miRNA)阵列)的 422 例 BC 样本和 31 例健康对照乳腺样本的 GSE57897 基因表达数据,并利用数据库与 CST7 mRNA 上具有保守靶点位置的 miRNA 进行比较。研究对象包括 60 个新鲜的 BC 组织样本和作为对照的邻近正常组织。本研究发现,与正常组织相比,CST7 和 hsa-miR-4793-5p 在肿瘤组织中的表达明显增加。本研究发现,与正常组织相比,CST7 和 hsa-miR-4793-5p 在肿瘤组织中明显增加,进一步分析表明,CST7 和 hsa-miR-4793-5p 基因表达改变之间存在明显关联。ROC曲线分析表明,CST7在BC肿瘤中的表达具有很高的准确性。不同分期和患者家族史之间的基因表达比较显示了重要的发现。由于这两个基因表达变化的灵敏度和特异性都很高,因此适合作为诊断和预后面板的一部分进行进一步研究。
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引用次数: 0
CCN5/WISP2 serum levels in patients with coronary artery disease and type 2 diabetes and its correlation with inflammation and insulin resistance; a machine learning approach 冠心病和 2 型糖尿病患者的 CCN5/WISP2 血清水平及其与炎症和胰岛素抵抗的相关性;一种机器学习方法
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-30 DOI: 10.1016/j.bbrep.2024.101857
Reza Afrisham , Vida Farrokhi , Seyed Mohammad Ayyoubzadeh , Akram Vatannejad , Reza Fadaei , Nariman Moradi , Yasaman Jadidi , Shaban Alizadeh

Introduction

Studies have shown various effects of CCN5/WISP2 on metabolic pathways, yet no prior investigation has established a link between its serum levels and CAD and/or T2DM. Therefore, this study seeks to explore the relation between CCN5 and the risk factor of CAD and/or diabetes, in comparison to individuals with good health, marking a pioneering endeavor in this field.

Methods

This case-control study investigates serum levels of CCN5, TNF-α, IL-6, adiponectin, and fasting insulin in a population of 160 individuals recruited into four equal groups (T2DM, CAD, CAD-T2DM, and healthy controls). Statistical tests comprise Chi-square tests, ANOVA, Spearman correlation, and logistic regression. ROC curves were used to represent the diagnostic potential of CCN5. Disease states are predicted by machine learning algorithms: Decision Tree, Gradient Boosted Trees, Random Forest, Naïve Bayes, and KNN. These models' performance was evaluated by various metrics, all of which were ensured to be robust by applying 10-fold cross-validation. Analyses were done in SPSS and GraphPad Prism and RapidMiner software.

Results

The CAD, T2DM, and CAD-T2DM groups had significantly higher CCN5 concentrations compared to the healthy control group (CAD: 336.87 ± 107.36 ng/mL, T2DM: 367.46 ± 102.15 ng/mL, CAD-T2DM: 404.68 ± 108.15 ng/mL, control: 205.62 ± 63.34 ng/mL; P < 0.001). A positive and significant correlation was observed between CCN5 and cytokines (IL-6 and TNF-α) in all patient groups (P < 0.05). Multinomial logistic regression analysis indicated a significant association between CCN5 and T2DM-CAD, T2DM, and CAD conditions (P < 0.001) even after adjusting for gender, BMI, and age (P < 0.001). Regarding the machine learning models, the Naïve Bayes model showed the best performance for classifying cases of T2DM, achieving an AUC value of 0.938±0.066. For predicting CAD, the Random Forest classifier achieved the highest AUC value of 0.994±0.020. In the case of CAD-T2DM prediction, the Naïve Bayes model demonstrated the highest AUC of 0.981±0.059, along with an Accuracy of 97.50 % ± 7.91 % and an F-measure of 96.67 % ± 10.54 %.

Conclusion

Our study has revealed, for the first time, a positive connection between CCN5 serum levels and the risk of developing T2DM and CAD. Nonetheless, more research is needed to ascertain whether CCN5 can serve as a predictive marker.
引言研究表明,CCN5/WISP2对代谢途径有各种影响,但之前的调查尚未确定其血清水平与CAD和/或T2DM之间的联系。方法本病例对照研究调查了 160 人血清中 CCN5、TNF-α、IL-6、脂肪连素和空腹胰岛素的水平,这些人被分为四个相同的组别(T2DM、CAD、CAD-T2DM 和健康对照组)。统计测试包括卡方检验、方差分析、斯皮尔曼相关性和逻辑回归。ROC 曲线用于表示 CCN5 的诊断潜力。疾病状态由机器学习算法预测:决策树、梯度提升树、随机森林、奈夫贝叶斯和 KNN。这些模型的性能通过各种指标进行评估,所有指标都通过应用 10 倍交叉验证来确保其稳健性。结果与健康对照组相比,CAD、T2DM 和 CAD-T2DM 组的 CCN5 浓度明显更高(CAD:336.87 ± 107.36 ng/mL;T2DM:367.46 ± 102.15 ng/mL;CAD-T2DM:404.68 ± 108.15 ng/mL;对照组:205.62 ± 63.34 ng/mL;P <;0.001)。在所有患者组中,CCN5 与细胞因子(IL-6 和 TNF-α)之间均存在明显的正相关性(P < 0.05)。多叉逻辑回归分析表明,即使调整了性别、体重指数和年龄,CCN5 与 T2DM-CAD、T2DM 和 CAD 病症之间仍存在显著关联(P <0.001)。在机器学习模型方面,Naïve Bayes 模型在对 T2DM 病例进行分类时表现最佳,其 AUC 值为 0.938±0.066。在预测 CAD 方面,随机森林分类器的 AUC 值最高,为 0.994±0.020。在预测 CAD-T2DM 时,Naïve Bayes 模型的 AUC 值最高,为 0.981±0.059,准确率为 97.50 % ± 7.91 %,F 值为 96.67 % ± 10.54 %。尽管如此,还需要更多的研究来确定CCN5是否可作为预测标志物。
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引用次数: 0
Characterization of antibodies induced by immunization of mice with isoglobotrihexosylceramide (iGb3) 用异球蛋白三己基甘油酰胺(iGb3)免疫小鼠所诱导抗体的特征
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-30 DOI: 10.1016/j.bbrep.2024.101855
Tetsuya Okuda
Isoglobotrihexosylceramide (iGb3), a well-characterized natural killer T cell ligand found in mammalian tissues, is also known as a glycosphingolipid that contains the human IgE epitope α-Gal (Galα1,3Gal) structure. Here, we analyzed the reactivity of several mice and human serum immunoglobulins against iGb3. Additionally, we isolated and characterized the variable region sequences of a monoclonal antibody that specifically recognizes iGb3. No IgE reactive with iGb3 was detected in sera from MRL/lpr mice, which are known to produce autoreactive antibodies, or in sera from healthy human donors. Furthermore, no induction of IgE and IgG was observed in the sera of mice immunized with iGb3; only IgM reactivity to iGb3 was detected. Further analysis of an anti-iGb3 monoclonal antibody generated from the splenocytes of an iGb3-immunized mouse revealed that the nucleotide sequences of the variable regions exhibited high homology to those of antibodies recognizing glycoconjugates containing Galα1,3 or Galα1,4 structures. These results indicate that the mouse genome harbors genes capable of encoding antibodies that recognize α-linked galactose-containing glycans, including iGb3, but that iGb3 is not sufficiently immunogenic to induce IgE in mammalian lymphocytes.
异球形三己糖基甘油三酯(iGb3)是哺乳动物组织中一种特征明显的自然杀伤 T 细胞配体,也被称为糖磷脂,含有人类 IgE 表位 α-Gal(Galα1,3Gal)结构。在这里,我们分析了几种小鼠和人类血清免疫球蛋白对 iGb3 的反应性。此外,我们还分离并鉴定了一种能特异性识别 iGb3 的单克隆抗体的可变区序列。在已知会产生自身反应性抗体的 MRL/lpr 小鼠血清和健康人类供体血清中均未检测到与 iGb3 反应的 IgE。此外,在用 iGb3 免疫的小鼠血清中也没有观察到 IgE 和 IgG 的诱导;只检测到了对 iGb3 的 IgM 反应性。对从免疫了 iGb3 的小鼠脾脏细胞中产生的抗 iGb3 单克隆抗体的进一步分析表明,可变区的核苷酸序列与识别含有 Galα1,3 或 Galα1,4 结构的糖结合物的抗体的核苷酸序列具有高度的同源性。这些结果表明,小鼠基因组中含有能够编码识别含α-连接半乳糖的聚糖(包括iGb3)的抗体的基因,但iGb3的免疫原性不足以诱导哺乳动物淋巴细胞产生IgE。
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引用次数: 0
LINC00261 triggers DNA damage via the miR-23a-3p/CELF2 axis to mitigate the malignant characteristics of 131I-resistant papillary thyroid carcinoma cells LINC00261通过miR-23a-3p/CELF2轴引发DNA损伤,从而减轻耐131I甲状腺乳头状癌细胞的恶性特征
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-30 DOI: 10.1016/j.bbrep.2024.101858
Qingyuan Tao , Xiaojin Li , Yanyan Xia , Bin Zheng , Yijun Yan , Songrun Wang , Li Jia

Background

Long-chain non-coding RNA (LINC00261) in the treatment of papillary thyroid carcinoma (PTC) with 131I is still unknown despite its proven anti-tumour effect in thyroid cancer (TC) and other types of cancer.

Methods

The database and RT-qPCR were used to analyze the expression level of LINC00261 in PTC and cell lines. PTC cells resistant to 131I (TPC-1/R) were created through ongoing exposure to a lethal dose of 131I, and a subcutaneous xenotransplantation model was developed using PTC mice. Bioinformatics analysis and dual-luciferase assays demonstrated the interaction between LINC00261, miR-23a-3p, and CELF2. RT-qPCR and Western blot were used to detect the expression of LINC00261, miR-23a-3p, and CELF2. Additionally, CCK-8, flow cytometry, immunofluorescence (IF), Western blot, and comet assay were employed to measure cell viability level and DNA damage.

Results

PTC cell lines exhibited a decrease in the expression of LINC00261. The growth and progression through the S-phase of TPC-1/R cells were suppressed by LINC00261, leading to increased apoptosis and DNA damage. The objective of LINC00261 was to regulate the axis of miR-23a-3p/CELF2. Downregulating LINC00261 enhances the growth and advancement of 131I-resistant cells in the S-phase by activating the miR-23a-3p/CELF2 pathway while suppressing cell death and DNA harm. The miR-23a-3p/CELF2 axis activates DNA damage in 131I-resistant PTC cells by LINC00261.

Conclusions

LINC00261 activates DNA damage in 131I-resistant PTC cells caused by miR-23a-3p/CELF2 axis, improving the progression of cancer cells of PTC.
背景长链非编码RNA(LINC00261)在甲状腺乳头状癌(PTC)131I治疗中的作用尚不清楚,尽管它在甲状腺癌(TC)和其他类型癌症中的抗肿瘤作用已得到证实。方法利用数据库和RT-qPCR分析LINC00261在PTC和细胞系中的表达水平。通过持续暴露于致死剂量的 131I 建立了对 131I 耐药的 PTC 细胞(TPC-1/R),并利用 PTC 小鼠建立了皮下异种移植模型。生物信息学分析和双荧光素酶测定证明了LINC00261、miR-23a-3p和CELF2之间的相互作用。RT-qPCR 和 Western 印迹被用来检测 LINC00261、miR-23a-3p 和 CELF2 的表达。此外,还采用了 CCK-8、流式细胞术、免疫荧光(IF)、Western 印迹和彗星试验来检测细胞活力水平和 DNA 损伤。LINC00261抑制了TPC-1/R细胞的生长和S期进展,导致细胞凋亡和DNA损伤增加。LINC00261 的目的是调节 miR-23a-3p/CELF2 轴。通过激活 miR-23a-3p/CELF2 通路,下调 LINC00261 可增强 131I 抗性细胞在 S 期的生长和进展,同时抑制细胞死亡和 DNA 损伤。结论LINC00261能激活miR-23a-3p/CELF2轴引起的131I耐药PTC细胞的DNA损伤,改善PTC癌细胞的进展。
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引用次数: 0
Integration analysis of cis- and trans-regulatory long non-coding RNAs associated with immune-related pathways in non-small cell lung cancer 非小细胞肺癌中与免疫相关通路有关的顺式和反式调控长非编码 RNA 的整合分析
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-28 DOI: 10.1016/j.bbrep.2024.101832
Yinqiang Liu , Hongjv Yang , Guoli Lv, Jin Duan, Wei Zhao, Yunfei Shi, Youming Lei

Background

Long non-coding RNAs (lncRNAs) are importantly involved in the initiation and progression of non-small cell lung cancer (NSCLC). However, the classification and mechanisms of lncRNAs remain largely elusive.

Aim

Hence, we addressed this through bioinformatics analysis.

Methods and results

We utilized microarray technology to analyze lncRNAs and mRNAs in twenty paired NSCLC tumor tissues and adjacent normal tissues. Gene set enrichment analysis, Kyoto Encyclopedia of Genes and Genomes, and Gene Ontology were conducted to discern the biological functions of identified differentially expressed transcripts. Additionally, networks of lncRNA-mRNA co-expression, including cis-regulation, lncRNA-transcription factor (TF)-mRNA, trans-regulation, and lncRNA-miRNA-mRNA interactions were explored. Furthermore, the study examined differentially expressed transcripts and their prognostic values in a large RNA-seq dataset of 1016 NSCLC tumors and normal tissues extracted from the Cancer Genome Atlas (TCGA). The analysis revealed 391 lncRNAs and 344 mRNAs with differential expression in NSCLC tumor tissues compared to adjacent normal tissues. Subsequently, 43,557 co-expressed lncRNA-mRNA pairs were identified, including 27 lncRNA-mRNA pairs in cis, 9 lncRNA-TF-mRNA networks, 34 lncRNA-mRNA pairs in trans, and 8701 lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks. Notably, these lncRNAs were found to be involved in immune-related pathways. Six significant transcripts, including NTF4, PTPRD-AS, ITGA11, HID1-AS1, RASGRF2-AS1, and TBX2-AS1, were identified within the ceRNA network and trans-regulation.

Conclusion

This study brings important insights into the regulatory roles of lncRNAs in NSCLC, providing a fresh perspective on lncRNA research in tumor biology.
背景长非编码RNA(lncRNA)在非小细胞肺癌(NSCLC)的发生和发展过程中起着重要作用。方法与结果我们利用芯片技术分析了20个配对的NSCLC肿瘤组织和邻近正常组织中的lncRNA和mRNA。我们利用基因组富集分析、京都基因和基因组百科全书以及基因本体论来鉴定差异表达转录本的生物学功能。此外,还探讨了lncRNA-mRNA共表达网络,包括顺式调控、lncRNA-转录因子(TF)-mRNA、反式调控和lncRNA-miRNA-mRNA相互作用。此外,研究还检测了从癌症基因组图谱(TCGA)中提取的1016个NSCLC肿瘤和正常组织的大型RNA-seq数据集中的差异表达转录本及其预后价值。分析发现,与邻近的正常组织相比,391个lncRNA和344个mRNA在NSCLC肿瘤组织中有差异表达。随后,共鉴定出43557对共同表达的lncRNA-mRNA,包括27对顺式lncRNA-mRNA、9个lncRNA-TF-mRNA网络、34对反式lncRNA-mRNA和8701个lncRNA-miRNA-mRNA竞争内源性RNA(ceRNA)网络。值得注意的是,这些lncRNA被发现参与了免疫相关通路。在ceRNA网络和反式调控中发现了6个重要的转录本,包括NTF4、PTPRD-AS、ITGA11、HID1-AS1、RASGRF2-AS1和TBX2-AS1。
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引用次数: 0
Anti-inflammatory effectiveness of Peperomia pellucida (L.) Kunth in rats induced with periodontitis Peperomia pellucida (L.) Kunth 对牙周炎大鼠的抗炎功效
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-26 DOI: 10.1016/j.bbrep.2024.101856
Dewi Lidya Ichwana Nasution , Sri Tjahajawati , Ratna Indriyanti , Amaliya Amaliya

Background

Periodontitis, marked by deep periodontal pocket depth (PPD), facilitates bacterial colonization and inflammation, necessitating adjunctive therapies. Although 0.2 % chlorhexidine (CHX) mouthwash is effective, its side effects have led to the search for alternative treatments. Peperomia pellucida (L.) Kunth, known as Pepper Elder, is a traditional medicinal plant with potential as an adjunctive herbal therapy for periodontitis.

Objective

This study aimed to investigate the anti-inflammatory efficacy of Peperomia pellucida extract in rats with induced periodontitis.

Methods

A post-test control group design was used in this laboratory experimental study. Four groups of Wistar strain Rattus norvegicus rats were utilized: a Pristine group (without periodontitis), a negative control group (induced periodontitis only), a positive control group (induced periodontitis and administered 0.2 % CHX), and an experimental group (induced periodontitis and administered 2.5 μL of Pepper Elder extract). Each treatment group received daily administration for one week. PPD measurements were taken on days 0, 3, 5, and 7. Blood serum was collected on day 7 for ELISA to measure IL-1β, TNF-α, IL-10, and IL-13 levels. Statistical analysis was performed using the Kruskal-Wallis test with a post hoc LSD and Mann-Whitney test.

Results

The extract-treated rats showed a decrease in PPD, with significant differences between the extract group and the negative control group (p < 0.05). TNF-α levels in the extract group differed significantly from the negative control group (p < 0.05) but not from the Pristine and positive control groups. IL-1β levels differed significantly only from the negative control group. IL-10 levels were significantly different from both the Pristine and negative control groups, while IL-13 levels differed significantly only from the negative control group.

Conclusion

Peperomia pellucida (L.) Kunth extract exhibits anti-inflammatory effects in rats with induced periodontitis.
背景牙周炎以牙周袋深度(PPD)为特征,有利于细菌定植和炎症,因此需要辅助治疗。虽然 0.2 % 洗必泰(CHX)漱口水很有效,但其副作用导致人们开始寻找替代疗法。本研究旨在探讨 Peperomia pellucida (L.) Kunth 提取物对诱导性牙周炎大鼠的抗炎功效。采用四组 Wistar 株 Rattus norvegicus 大鼠:原始组(无牙周炎)、阴性对照组(仅诱发牙周炎)、阳性对照组(诱发牙周炎并注射 0.2 % CHX)和实验组(诱发牙周炎并注射 2.5 μL 胡椒萃取物)。每个治疗组每天给药一周。在第 0、3、5 和 7 天测量 PPD。第 7 天采集血清进行 ELISA 检测,以测定 IL-1β、TNF-α、IL-10 和 IL-13 的水平。结果经提取物处理的大鼠 PPD 有所下降,提取物组与阴性对照组之间存在显著差异(p < 0.05)。提取物组的 TNF-α 水平与阴性对照组有显著差异(p < 0.05),但与原始对照组和阳性对照组无显著差异。IL-1β 水平仅与阴性对照组有显著差异。IL-10水平与原始组和阴性对照组均有显著差异,而IL-13水平仅与阴性对照组有显著差异。
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引用次数: 0
Recent advances in aptamer discovery, modification and improving performance 发现、修改和提高灵敏度的最新进展
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-24 DOI: 10.1016/j.bbrep.2024.101852
Arezoo Fallah , Abbas Ali Imani Fooladi , Seyed Asghar Havaei , Mahdieh Mahboobi , Hamid Sedighian
Aptamers are nucleic acid (Ribonucleic acid (RNA) and single strand deoxyribonucleic acid (ssDNA)) with a length of approximately 25–80 bases that can bind to particular target molecules, similar to monoclonal antibodies. Due to their many benefits, which include a long shelf life, minimal batch-to-batch variations, extremely low immunogenicity, the possibility of chemical modifications for improved stability, an extended serum half-life, and targeted delivery, they are receiving a lot of attention in a variety of clinical applications. The development of high-affinity modification approaches has attracted significant attention in aptamer applications. Stable three-dimensional aptamers that have undergone chemical modification can engage firmly with target proteins through improved non-covalent binding, potentially leading to hundreds of affinity improvements. This review demonstrates how cutting-edge methodologies for aptamer discovery are being developed to consistently and effectively construct high-performing aptamers that need less money and resources yet have a high chance of success. Also, High-affinity aptamer modification techniques were discussed.
适配体是长度约为 25-80 个碱基的核酸(核糖核酸 (RNA) 和单链脱氧核糖核酸 (ssDNA)),可以与特定的目标分子结合,类似于单克隆抗体。由于其具有保存期长、批次间差异小、免疫原性极低、可通过化学修饰提高稳定性、延长血清半衰期和靶向给药等诸多优点,因此在各种临床应用中受到广泛关注。高亲和性修饰方法的开发在适配体应用中备受关注。经过化学修饰的稳定三维适配体可以通过改进的非共价结合与目标蛋白质牢固结合,从而有可能提高数百种亲和力。这篇综述展示了如何开发最前沿的适配体发现方法,以持续有效地构建高性能的适配体,从而减少资金和资源的投入,同时又有很大的成功几率。此外,还讨论了高亲和性适配体修饰技术。
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Modifications in the C-terminal tail of TrkC significantly alter neurotrophin-3-promoted outgrowth of neurite-like processes from PC12 cells 对 TrkC C 端尾部的修饰会显著改变神经营养素-3 促进的 PC12 细胞神经样突起的生长过程
IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-10-23 DOI: 10.1016/j.bbrep.2024.101853
Pawel Krawczyk, Dagmara Klopotowska, Janusz Matuszyk
TrkB and TrkC are quite common neurotrophin receptors found on the same cells in CNS. In the C-terminal tail, TrkB and TrkC differ only in two amino acid residues at positions immediately preceding the tyrosine residue, which, upon phosphorylation, becomes the docking site for phospholipase Cγ1 (PLCγ1). The question arose whether such a difference near the PLCγ1 docking site might contribute to differential response to neurotrophin. PC12 clones with the following receptors were obtained: wild-type TrkC, TrkC-Y820F with a defective PLCγ1 binding site, TrkC-T817S–I819V with two amino acid residues replaced with those in the TrkB tail. The outgrowth of neurite-like processes from TrkC-Y820F-containing cells appeared to be impaired, while the TrkC-T817S–I819V variant appeared more effective than wild-type TrkC in promoting the outgrowth of neurite-like processes after neurotrophin stimulation, at least in the compared PC12 cell clones. Taken together, both the tyrosine residue at the PLCγ1 docking site and the amino acid residues immediately preceding it appear important for TrkC-supported outgrowth of neurite-like processes.
TrkB 和 TrkC 是相当常见的神经营养素受体,存在于中枢神经系统的相同细胞中。在 C 端尾部,TrkB 和 TrkC 仅在紧接酪氨酸残基之前的两个氨基酸残基上存在差异,酪氨酸残基在磷酸化后成为磷脂酶 Cγ1 (PLCγ1)的对接位点。由此产生的问题是,PLCγ1对接位点附近的这种差异是否会导致对神经营养素的不同反应。研究人员获得了具有以下受体的 PC12 克隆:野生型 TrkC、具有 PLCγ1 结合位点缺陷的 TrkC-Y820F、用 TrkB 尾部的两个氨基酸残基替换了两个氨基酸残基的 TrkC-T817S-I819V。含 TrkC-Y820F 细胞神经样突起的生长似乎受到了影响,而 TrkC-T817S-I819V 变体在神经营养素刺激后促进神经样突起生长方面似乎比野生型 TrkC 更有效,至少在比较的 PC12 细胞克隆中是如此。综上所述,PLCγ1对接位点的酪氨酸残基和紧接其前的氨基酸残基对于TrkC支持的神经样突起的生长似乎都很重要。
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