Experimental and Therapeutic Medicine
·
2020
·Vol. 20
·期 6
·1-1
·
IF 1.785
Tongtong Niu,Yan An,Tingting Lv,Dongning Liu
Diabetic retinopathy (DR) is a serious complication of diabetes and the most common metabolic disorder. Recently, long non‑coding (lnc)RNAs have been identified as critical regulators of DR. Ribosomal protein SA pseudogene 52 (RPSAP52) is an oncogenic lncRNA expressed in pituitary tumors. The present study aimed to investigate the functions of RPSAP52 in DR. RPSAP52 levels in the plasma of diabetic patients with or without DR complication was detected. Luciferase reporter assays, RT‑qPCR and western blotting were performed to detect the interaction between RPSAP52 and micro RNA (miR)‑365. Moreover, expression vectors of RPSAP52 and Timp3, as well as miR‑365 mimics were transfected into ARPE‑19 cells exposed to high glucose and the apoptotic cells were detected. The results showed that RPSAP52 was downregulated in patients with DR compared with patients with diabetes without obvious complications. RPSAP52 directly interacted with miR‑365, while overexpression of RPSAP52 and miR‑365 did not affect the expression of one another. In addition, overexpression of RPSAP52 upregulated TIMP metallopeptidase inhibitor 3 (Timp3) in retinal pigment epithelial (RPE) cells. High glucose treatment led to downregulated RPSAP52 and Timp3, but upregulated miR‑365 in RPE cells. Moreover, cell apoptosis analysis identified that overexpression of RPSAP52 and Timp3 led to a decreased apoptotic rate of RPE cells under high glucose treatment. Therefore, it was speculated that RPSAP52 may upregulate Timp3 by serving as the endogenous sponge of miR‑365 in DR to suppress RPE cell apoptosis.
研究领域:
分子生物学
·
医学遗传学
·
眼科学
·
糖尿病学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF APPLIED ENTOMOLOGY
·
2022
·Vol. 146
·期 7
·823-837
·
IF 2.183
Xuanyu He,Zhaodong Hao,Hui Li,Tian Xu,Ruixu Chen,Xiaohong Xia,Shouyin Li,Dejun Hao
Understanding the mechanism of insect resistance to toxic effects of phytochemicals can provide an insight into plant–herbivore interactions. Monochamus alternatus Hope, a main vector of pine wood nematode ( Bursaphelenchus xylophilus ), prefers to infest masson pine ( Pinus massoniana Lamb) and causes huge economic and environmental losses. α-Pinene is the primary component of resin produced by masson pine and plays a crucial role in defending against herbivorous insects, but its defensive effects on M . alternatus larvae are barely known. Here, we explored the physiological metabolism in M . alternatus larvae that were fumigated with (+)-α-pinene, (-)-α-pinene, β-pinene, (R)-(+)-limonene, myrcene and (-)-α-phellandrene. (+)-α-Pinene showed the highest fumigant toxicity to M . alternatus larvae among these monoterpenes, and the fourth instar larvae possessed the highest resistance to this compound at the larval stage. Comparative transcriptome analysis detected 423 up-regulated and 311 down-regulated genes in the larvae fumigated with (+)-α-pinene, which were mainly related to detoxification, energy and protein metabolism. Among the differentially expressed genes encoding for detoxification enzymes, 14 P450s, 13 UGTs, 1GSTs and 3ABCs were dramatically up-regulated in the larvae fumigated with (+)-α-pinene. Silencing of P450s via RNA interference led to an increased mortality in the larval fumigated with (+)-α-pinene. Piperonyl butoxide exposure significantly increased the mortality and inhibited the expressions of P450s, UGTs and ABCs in the larvae fumigated with (+)-α-pinene. These results suggested an important role of P450s in the resistance of M . alternatus larvae to (+)-α-pinene fumigation, which may facilitate the understanding of terpenoid detoxification in M . alternatus larvae.
研究领域:
昆虫学
·
分子生物学
·
植物-昆虫互作
✓ 已验证
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2015
·
IF 4.097
Huanshun Yin,Yunlei Zhou,Zhiqing Yang,Yunlong Guo,Xinxu Wang,Shiyun Ai,Xiansheng Zhang
N6-methyladenosine (m6A) is a ubiquitous modification in mRNA and other RNAs across most eukaryotes, which is likely dynamically regulated throughout developmental processes. In this work, an electrochemical immunosensor was fabricated for RNA methylation detection using gold nanoparticles modified glassy carbon electrode (AuNPs/GCE) as substrate electrode. The assembly of 4-aminophenylboronic acid on AuNPs/GCE triggered the further immobilization of anti-m6A antibody on the substrate electrode through the specific interaction between boric acid and glycosyl of antibody, which led to the following capture of methylated RNA through the immunoreaction. A decreased electrochemical reduction signal of the redox probe of [Fe(CN) 6 ] 3− was obtained based on the synergistic effect of electrostatic repulsion and steric hindrance. The developed method showed a wide linear range for methylated RNA from 0.01 to 10 nM and the detection limit was 2.57 pM (S/N = 3). Moreover, the down-regulated expression of methylation level of mRNA in the leaves of rice seedlings after incubation with 6-benzylaminopurine was confirmed by the fabricated biosensor, indicating the great potential for assaying the expression level of methylated RNA in real sample.
研究领域:
生物化学
·
分析化学
·
分子生物学
·
植物科学
✓ 已验证
产品关联
DOI
高影响因子
METABOLIC BRAIN DISEASE
·
2021
·Vol. 36
·期 7
·2059-2069
·
IF 3.584
Yang Ling,Wang Lijuan,Wang Juan,Liu Ping
Long non-coding RNAs (lncRNAs) play important roles in ischaemic stroke. This study aimed to investigate the role and potential mechanism of lncRNA Gm11974 in ischaemic stroke. Mouse neuroblastoma N2a cells were treated with oxygen–glucose deprivation (OGD). The levels of Gm11974, microRNA-122-5p (miR-122-5p) and semaphorin 3A (SEMA3A) were detected by quantitative real-time PCR (qRT-PCR) or western blot. Cell viability and apoptosis were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, Caspase-3 Assay Kit and flow Cytometry. The levels of oxidative stress indicators were measured by using commercial kits. The relationship between miR-122-5p and Gm11974 or SEMA3A was verified by dual-luciferase reporter, RNA immunoprecipitation and RNA pull-down assays. Middle cerebral artery occlusion (MCAO) in mice was used to mimic ischaemic stroke. Gm11974 and SEMA3A were up-regulated, while miR-122-5p was down-regulated in OGD-treated N2a cells and MCAO mice. Down-regulation of Gm11974 ameliorated OGD-mediated N2a cell damage by increasing cell viability and reducing cell apoptosis and oxidative stress. Gm11974 promoted OGD-induced injury in N2a cells via negatively regulating miR-122-5p. Also, miR-122-5p alleviated OGD-resulted N2a cell injury by targeting SEMA3A. Moreover, silencing of Gm11974 decreased infarct volume and neurological score in MCAO mice. Knockdown of Gm11974 attenuated neuronal injury in ischaemic stroke by regulating miR-122-5p/SEMA3A signaling pathway.
研究领域:
神经科学
·
分子生物学
·
细胞生物学
·
病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Yanming Lai,Wei Xiao,Zhiwei Lei,Huiping Long,Jiting Deng,Yiyu Wang,Lu Tao,Xingxing Liu,Jinjun Wu,Qiwei Zhang,Donglin Cao,Heng Xiao
Exosomal microRNAs (miRNAs) serve as dependable and noninvasive biomarkers for early cancer diagnosis. However, the accurate and feasible detection of exosomal miRNAs is often hindered by their low abundance and the requirement of specialized equipment for miRNA detection. In this study, we present a novel approach, termed primer exchange reaction-based fluorescence emission with cascade RNA aptamers transcription (PERFECT) for the highly sensitive detection of exosomal miRNA. The design of this method involves the selective interaction of a DNA probe with the target miRNA, leading to its activation. Once activated, isothermal signal amplification and RNA aptamer transcription are initiated, resulting in an amplified fluorescent signal within 90 min. This method achieves a detection limit of 2.2 fM at 37 °C and 2.7 fM at room temperature (25 °C). We used the PERFECT technology to analyze miR-21 expression levels in cell extracts, cell-derived exosomes, and human plasma-derived exosomes, achieving a diagnostic accuracy of 93.6% in distinguishing hepatocellular carcinoma (HCC) patients. Overall, this study highlights its broad range of detection temperature, simplicity of the detection process, and strong potential for clinical application, rendering it a promising tool for early cancer diagnosis.
研究领域:
分子生物学
·
医学诊断
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Chemical Biology
·
2019
·Vol. 15
·期 11
·1110-1119
·
IF 12.154
Wang Pengchong,Tang Wei,Li Zeyao,Zou Zhongyu,Zhou Ying,Li Ran,Xiong Tianyu,Wang Jianbin,Zou Peng
RNA molecules are highly compartmentalized in eukaryotic cells, with their localizations intimately linked to their functions. Despite the importance of RNA targeting, our current knowledge of the spatial organization of the transcriptome has been limited by a lack of analytical tools. In this study, we develop a chemical biology approach to label RNAs in live cells with high spatial specificity. Our method, called CAP-seq, capitalizes on light-activated, proximity-dependent photo-oxidation of RNA nucleobases, which could be subsequently enriched via affinity purification and identified by high-throughput sequencing. Using this technique, we investigate the local transcriptomes that are proximal to various subcellular compartments, including the endoplasmic reticulum and mitochondria. We discover that messenger RNAs encoding for ribosomal proteins and oxidative phosphorylation pathway proteins are highly enriched at the outer mitochondrial membrane. Due to its specificity and ease of use, CAP-seq is a generally applicable technique to investigate the spatial transcriptome in many biological systems.
研究领域:
分子生物学
·
细胞生物学
·
转录组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANTI-CANCER DRUGS
·
2021
·Vol. 33
·期 1
·e145
·
IF 2.248
Liu Dan,Wang Yaya,Li Haiming,Peng Sida,Tan Huo,Huang Zhenqian
Multiple myeloma is a frequent hematologic malignancy. Bortezomib is the first-line drug for multiple myeloma chemotherapy. The present study aimed to investigate the potential role and mechanism of circular RNA chaperonin containing TCP1 subunit 3 (circ-CCT3) in bortezomib resistance of multiple myeloma . The levels of circ-CCT3, microRNA-223-3p ( miR-223-3p ), and bromodomain-containing 4 (BRD4) were detected by quantitative real-time PCR or western blot. Cell Counting Kit-8 (CCK-8) method was used to measure the half-inhibitory concentration of bortezomib and cell viability. Cell cycle distribution, apoptosis, proliferation and migration were determined by flow cytometry, 5-ethynyl-2′-deoxyuridine, and wound healing assay. The levels of relevant proteins were checked via western blot. The binding association between miR-223-3p and circ-CCT3/BRD4 was validated via a dual-luciferase reporter assay. Circ-CCT3 and BRD4 were upregulated, while miR-223-3p was downregulated in bortezomib -resistant multiple myeloma patients and cells. Silencing of circ-CCT3 enhanced the sensitivity of bortezomib -resistant multiple myeloma cells to bortezomib . Circ-CCT3 knockdown weakened bortezomib resistance via modulating miR-223-3p . Moreover, miR-223-3p increased bortezomib sensitivity by inhibiting BRD4. Downregulation of circ-CCT3 attenuated bortezomib resistance of multiple myeloma via regulating miR-223-3p /BRD4 pathway, which provided a new potential target for multiple myeloma chemoresistance.
研究领域:
肿瘤学
·
分子生物学
·
血液学
✓ 已验证
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2020
·
IF 2.751
Benshui Shu,Haikuo Yu,Yuning Li,Hongxin Zhong,Xiangli Li,Liang Cao,Jintian Lin
The fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae) is a polyphagous pest with 353 plant species as its hosts, including maize, sorghum, cotton, and rice. Azadirachtin is one of the most effective botanical insecticides . The effect of azadirachtin against S. frugiperda remains to be determined. Here we report strong growth inhibition of azadirachtin on S. frugiperda larvae under either 1.0 or 5.0 μg/g azadirachtin. To explore the relevant mechanisms, the larvae fed with normal artificial diet and with 1.0 μg/g azadirachtin exposure for 3 days were collected as samples for RNA-Seq. RNA-Seq on S. frugiperda larvae under different treatments identified a total of 24,153 unigenes , including 3494 novel genes, were identified. Among them, 1282 genes were affected by 1.0 μg/g azadirachtin exposure, with 672 up-regulated and 610 down-regulated. The impacted genes include 61 coding for detoxification enzymes (31 P450s , 7 GSTs , 11 CarEs , 7 UGTs and 5 ABC transporters), 31 for cuticle proteins, and several proteins involved in insect chitin and hormone biosynthesis . Our results indicated that azadirachtin could regulate the growth of S. frugiperda by affecting insect chitin and hormone biosynthesis pathway. The enhanced expression of detoxification enzymes might be related to detoxifying azadirachtin. These findings provided a foundation for further delineating the molecular mechanism of growth regulation induced by azadirachtin in S. frugiperda larvae.
研究领域:
昆虫学
·
分子生物学
·
农药科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Acta Pharmaceutica Sinica B
·
2024
·
IF 14.7
Li Qiao,Zhiyao Li,Bowen Li,Fu Zhang,Zhuo Yao,Chongzhi Wu,Honglin Tang,Qi Pan,Peihua Shi,Yuan Ping
Osteoarthritis (OA) is a type of highly prevalent heterogeneous degenerative disease that leads to joint pain, deformity, the destruction of articular cartilage, and eventual disability. The current treatment strategies for OA often suffer from systemic side effects, poor anti-inflammatory efficacy, and persistent pain. To address these issues, we develop light-inducible nanomedicine that enables the co-delivery of anti-inflammatory drug (diacerein, DIA) and small interfering RNA (siRNA) targeting nerve growth factor (NGF) for pain relief to enhance the therapeutic efficacy of OA. The nanomedicine is based on poly( β -amino-ester)-coated gold nanocages (AuNCs), which is further incorporated with the phase-change material (lauric acid/stearic acid, LA/SA). Following intra-articular (IA) injection in vivo , the nanomedicine displays high degree of drug accumulation and retention in the joint lesion of OA mouse models. The photothermal effect, induced by AuNCs, not only promotes DIA and siRNA release, but also upregulates the expression of heat shock protein 70 (HSP-70) to resist the apoptosis of chondrocytes in the inflammatory condition. The internalization of both DIA and siRNA results in strong anti-inflammatory and pain-relieving effects, which greatly contribute to the joint repair of OA mice. This study offers a promising combination strategy for OA treatment.
研究领域:
纳米医学
·
药物递送系统
·
骨科
·
生物医学工程
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food & Function
·
2024
·
IF 5.1
Jingyi Ren,Jiarui Liu,Jinshi Zuo,Zhenao Zhang,Dan Huang,Xuanyi Liu,Miaomiao Lu,Yadong Zhang,Yang Su,Yuxia Ma
Background: Perfluorooctane sulfonate (PFOS) is a persistent, widely present environmental pollutant, and its toxicity to male reproduction has gradually attracted attention. Flaxseed oil (FO) is a dietary oil abundant in α-linolenic acid and has been demonstrated to possess multiple health benefits. However, whether FO protects against PFOS-induced testicular injury and its mechanism remain unclear. Methods: C57/BL6 mice were gavaged with different concentrations of FO or PFOS (10 mg kg−1) for 28 days. Blood and testicular tissues were collected for histopathology, proteomics, and biochemical and molecular analyses. Results: Our results showed that FO supplementation significantly attenuated PFOS-induced testicular injury, as indicated by histopathological changes, decreased oxidative stress level, increased sperm count, decreased rate of sperm malformation, and improved functional markers of spermatogenesis. Proteomic analysis showed that differentially expressed proteins were notably enriched in spliceosome pathways. Machine learning algorithms were used to screen the hub gene, and PRPF3 and PUF60 proteins were found to be important for FO to exert protective benefits to testicular injury. Western blot results confirmed that FO supplementation could increase the protein expression of PRPF3 and decrease the protein expression of PUF60 in PFOS-exposed mice. Conclusions: This study revealed that FO can alleviate PFOS-induced testicular dysfunction by regulating RNA alternative splicing. The spliceosome-related proteins PRPF3 and PUF60 may be the potential targets for FO to alleviate PFOS-induced testicular injury. FO supplementation may be an effective dietary intervention to prevent adverse effects of PFOS on testes.
研究领域:
毒理学
·
生殖生物学
·
营养科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yusi Li,Fang Yang,Shunmei Li,Ruo Yuan,Yun Xiang
Proteins play vital roles in regulating a series of living activities and can be potentially considered to be useful indicators for diagnosing various diseases. Here, a target-triggered transcription cascade amplification strategy with the use of the fluorescence switch-on RNA aptamer is constructed for label-free and sensitive platelet-derived growth factor-BB (PDGF-BB) protein biomarker detection. The target molecule of PDGF-BB binds a rationally designed multifunctional hairpin probe to switch its structure and to expose a primer binding region. The primer sequence further binds such a region to initiate the target recycling and lighting-up RNA aptamer transcription cascade cycles to yield tremendous RNA aptamers. And, the fluorescence of the organic dye is substantially enhanced upon binding to these RNA aptamers for realizing highly sensitive and label-free PDGF-BB detection with the detection limit being lowered to 0.8 pM. Moreover, the developed strategy has superior selectivity and exhibits a promising potential to detect PDGF-BB spiked into serums with buffer dilution, which makes this method an attractive sensing system for detecting other biomarkers at trace levels.
研究领域:
生物化学
·
分子生物学
·
医学诊断
✓ 已验证
DOI
PubMed
高影响因子
Viruses-Basel
·
2019
·Vol. 11
·期 4
·314
·
IF 3.811
Gaobo Zhang,Jian Yang,Fujun Qin,Congrui Xu,Jia Wang,Chengfeng Lei,Jia Hu,Xiulian Sun
Dendrolimus punctatuscypovirus (DpCPV), belonging to the genusCypoviruswithin the familyReoviridae, is considered the most destructive pest of pine forests worldwide. DpCPV has a genome consisting of 10 linear double-stranded RNA segments. To establish a reverse genetics system, we cloned cDNAs encoding the 10 genomic segments of DpCPV into three reverse genetics vectors in which each segment was transcribed under the control of a T7 RNA polymerase promoter and terminator tagged with a hepatitis delta virus ribozyme sequence. We also constructed avp80-knockoutAutographa californicamultiple nucleopolyhedrovirus bacmid to express a T7 RNA polymerase codon-optimized for Sf9 cells. Following transfection of Sf9 cells with the three vectors and the bacmid, occlusion bodies (OBs) with the typical morphology of cypovirus polyhedra were observed by optical microscopy. The rescue system was verified by incorporation of aHindIII restriction enzyme site null mutant of the 9th genomic segment. Furthermore, when we co-transfected Sf9 cells with the reverse genetics vectors, the bacmid, and an additional vector bearing anegfpgene flanked with the 5′ and 3′ untranslated regions of the 10th genomic segment, aggregated green fluorescence co-localizing with the OBs was observed. The rescued OBs were able to infectSpodopetra exigualarvae, although their infectivity was significantly lower than that of wild-type DpCPV. This reverse genetics system for DpCPV could be used to explore viral replication and pathogenesis and to facilitate the development of novel bio-insecticides and expression systems for exogenous proteins.Keywords:DpCPV;reverse genetics system;vp80-knockout;foreign protein
研究领域:
病毒学
·
分子生物学
·
昆虫学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）