BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Jiahao Li,Xuping Wu,Yue Li,Xin Wang,Huachuan Huang,Dan Jian,Yanke Shan,Yue Zhang,Chengcheng Wu,Guolei Tan,Shouyu Wang,Fei Liu
Classical gold standard HBV detection relies on expensive devices and complicated procedures, thus is always restricted in large-scale hospitals and centers for disease control and prevention. To extend HBV detection to primary clinics especially in underdeveloped areas, we design amplification-free smartphone-based attomolar HBV detecting technique based on single molecule sensing. Verified by synthesized HBV target DNA , this technique reaches a detection limit at attomolar concentration (100 aM); and verified by 110 clinical samples, it also reaches a rather high sensitivity of 10 4 copy/mL (≈2000 IU/mL) with a high accuracy of 93.64% certificated by gold standard HBV detecting devices. Besides, this technique can quantify HBV viral load in 70 min only using portable and inexpensive devices as well as simple operations. Because of its cost-effective, field-portable and operable design, highly sensitive and selective detecting capability and wireless data connectivity, this technique can be potentially used in mobile HBV diagnoses and share HBV epidemic information especially in resource limited situations.
研究领域:
医学检验
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分子生物学
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DOI
PubMed
高影响因子
Nutrients
·
2024
·Vol. 16
·期 5
·600
·
IF 4.8
Juan Yang,He Gao,Tiantian Zhang,Yong Fan,Yuwei Wu,Xinyu Zhao,Ying Li,Lei Wu,Hui Zhao,Lingshuang Yang,Haojie Zhong,Longyan Li,Xinqiang Xie,Qingping Wu
Hepatocellular carcinoma (HCC), being ranked as the top fifth most prevalent cancer globally, poses a significant health challenge, with a considerable mortality rate. Hepatitis B virus (HBV) infection stands as the primary factor contributing to HCC, presenting substantial challenges in its treatment. This study aimed to identify lactic acid bacteria (LAB) with anti-HBV properties and evaluate their impact on the intestinal flora in HBV-associated HCC. Initially, two LAB strains,Levilactobacillus brevisSR52-2 (L. brevisSR52-2) andLeviLactobacillus delbrueckiisubsp.bulgaicusQ80 (L. delbrueckiiQ80), exhibiting anti-HBV effects, were screened in vitro from a pool of 498 LAB strains through cell experiments, with extracellular expression levels of 0.58 ± 0.05 and 0.65 ± 0.03, respectively. These strains exhibited the capability of inhibiting the expression of HBeAg and HBsAg. Subsequent in vitro fermentation, conducted under simulated anaerobic conditions mimicking the colon environment, revealed a decrease in pH levels in both the health control (HC) and HCC groups influenced by LAB, with a more pronounced effect observed in the HC group. Additionally, the density of total short-chain fatty acids (SCFAs) significantly increased (p< 0.05) in the HCC group. Analysis of 16S rRNA highlighted differences in the gut microbiota (GM) community structure in cultures treated withL. brevisSR52-2 andL. delbrueckiiQ80. Fecal microflora in normal samples exhibited greater diversity compared to HBV-HCC samples. The HCC group treated with LAB showed a significant increase in the abundance of the phyla Firmicutes, Bacteroidetes and Actinobacteria, while Proteobacteria significantly decreased compared to the untreated HCC group after 48 h. In conclusion, the findings indicate that LAB, specificallyL. brevisSR52-2 andL. delbrueckiiQ80, possessing antiviral properties, contribute to an improvement in gastrointestinal health.
研究领域:
微生物学
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医学研究
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癌症生物学
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病毒学
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胃肠病学
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DOI
PubMed
高影响因子
MEDICINAL CHEMISTRY RESEARCH
·
2023
·Vol. 32
·期 5
·899-909
·
IF 2.6
Ye Zhen,Zhao Tong-Shi-Yao,Li Shan-Bin,Zhou Xian-Li,Luo Qin,Qin Jiang-Ke,Liang Cheng-Qin,Wang Ping,Ge Guang-Bo
Previous in vivo and in vitro studies revealed that esculetin (Fig. 1 ) has anti-hepatitis B virus (anti-HBV) activity as well as a protective effect on liver damage caused by duck hepatitis B virus. We designed and synthesized a series of esculetin derivatives, introduced side chains containing various amino groups into site 7 of the parent structure, and synthesized C-4 and C-8 substituted derivatives with the goal of investigating their anti-HBV activities. In vitro anti-HBV activity was performed against HepG2.2.15 cells by using Enzyme-Linked Immunosorbent Assay(ELISA) kit and cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay with lamivudine as the positive control. The results demonstrated that several compounds showed moderate anti-HBV activity, while the introduction of morpholine groups could significantly inhibit the expression of hepatitis B e antigen (HBeAg) and the introduction of the 2-methylimidazole group could significantly inhibit the expression of Hepatitis B surface antigen (HBsAg). Among all tested compounds, compound 4a demonstrated the best anti-HBeAg activity (IC 50 = 15.8 ± 4.2 μM), while compound 6d demonstrated the best anti-HBsAg activity (IC 50 = 21.4 ± 2.8 μM). Compounds 6b and 6c showed moderate anti-HBV activity and HBsAg inhibition. Compounds 4b showed moderate anti-HBV activity and an inhibitory effect on HBeAg. In addition, compounds 4a , 4c , 4d , 6b , 6c and 6d showed improved metabolic stability. This study provides useful guidance for the discovery of anti-HBV drugs, which merits further investigation. Graphical Abstract
研究领域:
药理学
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病毒学
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DOI
PubMed
高影响因子
Autophagy
·
2016
·
IF 9.108
Na Xie,Kefei Yuan,Li Zhou,Kui Wang,Hai-Ning Chen,Yunlong Lei,Jiang Lan,Qinqin Pu,Wei Gao,Lu Zhang,Guobo Shen,Qifu Li,Hengyi Xiao,Hong Tang,Rong Xiang,Mingliang He,Pinghui Feng,Edouard C. Nice,Yuquan Wei,Haiyuan Zhang,Jiayin Yang,Canhua Huang
Adenosine monophosphate-activated protein kinase (AMPK) is a crucial energy sensor that maintains cellular energy homeostasis. AMPK plays a critical role in macroautophagy/autophagy, and autophagy facilitates hepatitis B virus (HBV) replication. To date, the intrinsic link among AMPK, autophagy and HBV production remains to be elucidated. Here, we demonstrate that PRKAA (a catalytic subunit of AMPK) is activated in response to HBV-induced oxidative stress, which in turn decreases the production of HBV. Mechanistic studies reveal that the autophagy machinery is associated with the inhibitory effect of PRKAA/AMPK on HBV production. Activation of PRKAA/AMPK promotes autolysosome-dependent degradation through stimulation of cellular ATP levels, which then leads to the depletion of autophagic vacuoles. Taken together, our data suggest that the activation of AMPK might be a stress response of host cells to restrict virus production through promotion of autophagic degradation. These findings therefore indicate that AMPK could provide a potential therapeutic target for HBV infection.
研究领域:
病毒学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
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2022
·
IF 4.117
Tan Qin,Yu Wang,Jing Nie,Lushan Yu,Su Zeng
Background Sodium taurocholate co -transportering polypeptide (NTCP, SLC10A1 ) is a vital bile acid transporter and the functional receptor of hepatitis B and D virus. The oligomerization of NTCP is important for the structural study of its interaction with HBV preS1 peptide. Methods Recombinant NTCPs were expressed in Sf9 host cell using baculoviruses. Function of recombinant NTCP was verified by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method. A quantitative fluorescence resonance energy transfer (FRET) method was established to analyze the interaction between NTCP wild type (WT) and mutants. Co-immunoprecipitation (Co-IP) was used to test the interaction between NTCP variants. Results Sub-cellular location of recombinant NTCPs varies with the modification of NTCP. Bands of monomer, dimer and oligomers were shown in gel analysis of NTCP. Significant FRET was observed between cyan florescence protein (CFP) tagged NTCP and yellow florescence protein (YFP) tagged NTCP. FRET efficiency between CFP- and YFP-NTCP S267F mutants was lower than WT. Co-IP results showed that S267F interacts with WT NTCP when co-expressed in cell. Conclusion Dimer is the predominant form of NTCP expressed in Sf9 when solubilized with detergent. FRET and Co-IP analysis support that NTCP forms oligomers in Sf9 cell. General significance Our results showed that NTCP formed oligomers in Sf9 cell. Meanwhile the FRET analysis of NTCP variants further elucidated the molecular mechanism of NTCP oligomerization.
研究领域:
病毒学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2017
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IF 3.255
Bingchen Ouyang,Fang Zhou,Le Zhen,Ying Peng,Jianguo Sun,Qianying Chen,Xiaoliang Jin,Guangji Wang,Jingwei Zhang
Tenofovir (TFV), a first-line anti-viral agent, has been prepared as various forms of prodrugs for better bioavailability, lower systemic exposure and higher target cells loading of TFV to enhance efficacy and reduce toxicity. TFV undergoes intracellular phosphorylation to form TFV diphosphate (TFV-DP) in target cell to inhibit viral DNA replication. Hence, TFV-DP is the key active metabolite that exhibits anti-virus activity, its intracellular exposure and half-life determine the final activity. Therefore, simultaneous monitoring prodrug, TFV and TFV-DP in target cells will comprehensively evaluate TFV prodrugs, both considering the stability of ester prodrug, and the intracellular exposure of TFV-DP. Thus we intended to develop a convenient general analytical method, taking tenofovir alafenamide (TAF) as a representative of TFV prodrugs. A sensitive LC–MS/MS method was developed, and TAF, TFV and TFV-DP were separated on a XSelect HSS T3 column (4.6 mm × 150 mm, 3.5 μm, Waters) with gradient elution after protein precipitation. The method provided good linearity for all the compounds (2–500 nM for TFV and TAF; 20–5000 nM for TFV-DP) with the correlation coefficients ( r ) greater than 0.999. Intra- and inter-day accuracies (in terms of relative error, RE < 10.4%) and precisions (in terms of coefficient of variation, CV < 14.1%) satisfied the standard of validation. The matrix effect, recovery and stability were also within acceptable criteria. Finally, we investigated the intracellular pharmacokinetics of TAF and its active metabolites in HepG2.2.15 cells with this method.
研究领域:
药理学
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分析化学
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病毒学
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DOI
高影响因子
ANALYST
·
2022
·Vol. 147
·期 24
·5623-5632
·
IF 5.227
Jiahai Wang,Le Luo,Yunhui Li,Lanfang Chen,Cenlin Gui,Jianji Zhu,Huizhen Li,Wenlong Wang,Daqi Chen
Solid-state nanopores have been proven as a powerful platform for label-free single-molecule analysis. However, due to its relatively low resolution and selectivity, developing biosensors with good translocation signals faces two significant challenges: (1) small-sized chemical or biological targets show difficulty in producing recognizable translocation signals because of their weak interaction with the nanopore and (2) protein interferents that widely exist in biological samples or buffers would considerably deteriorate the noise level of the nanopore, submerging the translocation signal. Herein, we demonstrate an effective way to overcome both the challenges. DNA cubes were used as signal transducers that can achieve an ultra-high (>50 : 1) signal-to-noise ratio (SNR) translocation signal, which is maintained even in protein interferent-rich buffers. A sensing strategy was constructed via hepatitis B virus (HBV) target-triggered cleavage of the component elements of the DNA cube with the assistance of the CRISPR–Cas12a technology, which caused a great drop in the translocation rate. The elements to cleave were optimized, and the sensor performance was tested in different protein stabilizer-rich buffers and human serum. Coupling with the polymerase chain reaction (PCR) pre-amplification technology, HBV-positive or -negative classification was achieved with the detection limit reaching 5 aM. It is worth noting that in our method, all reaction buffers were directly used without further optimization, which is of great help for the practical application of solid-state nanopores.
研究领域:
生物技术
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纳米技术
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分子生物学
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病毒学
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DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
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2022
·Vol. 20
·期 1
·1-16
·
IF 9.429
Wang Dan,Chen Ling,Li Chengbi,Long Quanxin,Yang Qing,Huang Ailong,Tang Hua
Background Currently, there are no curative drugs for hepatitis B virus (HBV). Complete elimination of HBV covalently closed circular DNA (cccDNA) is key to the complete cure of hepatitis B virus infection. The CRISPR/Cas9 system can directly destroy HBV cccDNA. However, a CRISPR/Cas9 delivery system with low immunogenicity and high efficiency has not yet been established. Moreover, effective implementation of precise remote spatiotemporal operations in CRISPR/Cas9 is a major limitation.Results In this work, we designed NIR-responsive biomimetic nanoparticles (UCNPs-Cas9@CM), which could effectively deliver Cas9 RNP to achieve effective genome editing for HBV therapy. HBsAg, HBeAg, HBV pgRNA and HBV DNA along with cccDNA in HBV-infected cells were found to be inhibited. These findings were confirmed in HBV-Tg mice, which did not exhibit significant cytotoxicity and minimal off-target DNA damage.Conclusion sThe UCNPs-based biomimetic nanoplatforms achieved the inhibition of HBV replication via CRISPR therapy and it is a potential system for efficient treatment of human HBV diseases.Graphical Abstract
研究领域:
分子生物学
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病毒学
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纳米技术
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治疗学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Hai-Bo Wang,Zi-Tao Zhong,Ting Zhang,Yuan-Di Zhao
Hepatitis B virus (HBV) can cause liver cancer, thus the detection of HBV-DNA is of great significance for liver cancer prevention. Since the concentration of HBV-DNA is often low, it is important to establish a highly sensitive detection method. In this study, microplate amplification and hybridization chain reaction (HCR) were used to improve the sensitive detection of HBV-DNA on dual strip biosensors. For the microplate-based amplification strategy on RS strip, the dual-DNA-AuNP probe could be immobilized on SA-Biotin-Anti-HBV-DNA-1 covered microplate through HBV-DNA. By adding dithiothreitol (DTT), a large amount of HS-RS-DNA on the dual-DNA-AuNP probe was dissociated. The indirect detection of HS-RS-DNA on RS strip could achieve the signal amplification of HBV-DNA. For the HCR-based strategy on HCR strip, HBV-DNA triggered the cascade reaction of H1-AuNP probe and H2-Biotin, a large amount of HCR product (AuNP-H1-H2-Biotin-AuNP-H1…) was produced. Thus, a lot of AuNP was fixed on the test zone of HCR strip to improve the HBV-DNA detection sensitivity. The detection limits by HCR strategy on HCR strip and microplate strategy on RS strip could reach to 5 pM and 0.1 nM, respectively, which were 200 and 10-fold higher than that of unamplified strategy, respectively. The recovery of HBV-DNA ranged from 97.2 to 102 % in the spiked human serum, presented a good accuracy for HBV-DNA detection in actual samples.
研究领域:
生物技术
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分子生物学
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医学诊断
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DOI
高影响因子
JOURNAL OF MEDICAL VIROLOGY
·
2026
·Vol. 98
·期 1
·e70777
·
IF 4.6
Lingzhu Zhao,Siduo Xu,Shouhan Yao,Zhiqiang Wei,Guohua Lou,Jinjin Qi,Haofeng Xu,Xueyu Wang,Zhenggang Yang,Min Zheng
The development of therapeutic strategies capable of achieving functional cure remains an unmet need in chronic hepatitis B management. Class A capsid assembly modulators (CAM-As) emerge as a promising treatment option. CAM-As not only directly disrupt the normal assembly of capsids, but some of which have also been reported to activate the innate immune response in animal models with unclear mechanisms. GLS4 is one of CAM-As with great potential. In this study, we investigate its capacity to activate immune response both in vitro and in vivo, as well as the underlying mechanisms involved. GLS4 activates the RIG-I-mediated interferon signaling pathway in both HBV-expressing hepatocellular carcinoma cell lines and HBV carrier mouse models, as indicated by RNA-seq. Besides, combination treatment with GLS4 and ritonavir elevates the frequencies of both peripheral blood IFNγ + NK cells and liver-resident IFNγ + CD8+ T cells in the pAAV/HBV1.2 hydrodynamic injection (HDI) model. In conclusion, our study reveals a previously unknown mechanism by which GLS4 activates the interferon signaling pathway in HBV-expressing hepatocytes. Furthermore, GLS4 partially restores innate and adaptive immunity in vivo. This signifies a potentially effective strategy for achieving a functional cure for HBV infection.
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PubMed
高影响因子
RSC Advances
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2017
·Vol. 7
·期 14
·8178-8185
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IF 3.108
Xueyang Fang,Xianlin Wu,Chang'e Li,Binwei Zhou,Xiaoyin Chen,Tianfeng Chen,Fang Yang
As a traditional Chinese medicine, baicalin exhibits high antitumor activity towards hepatitis B virus (HBV)-infected liver cancer, but low toxicity towards normal tissues. However, the properties of baicalin, including its low hydrophilicity and poor biocompatibility, have limited its clinical application. Herein, in this study, we have designed and synthesized multifunctional selenium nanoparticles (SeNPs) with baicalin (B) and folic acid (FA) surface-modifications for the targeted treatment of HBV infected liver cancer. B–SeNPs–FA showed appropriate particle size distribution, high stability, and higher cellular uptake by tumor cells than normal cells. The nanoparticles primarily targeted lysosomes in HepG2215 cells through utilization of the main pathways of caveolae-mediated endocytosis and energy-dependent endocytosis. B–SeNPs–FA induced HepG2215 cell apoptosis by down-regulating the generation of reactive oxygen species (ROS) and the expression of the HBxAg protein. In addition, B–SeNPs–FA exhibited an excellent ability to inhibit cancer cell migration and invasion. Taken together, these results suggest that baicalin-loaded selenium nanoparticles with a folic acid targeting moiety could be a promising strategy for the design and synthesis of cancer-targeted nanomaterials to treat HBV-infected liver cancer.
研究领域:
药理学
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肿瘤学
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纳米医学
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DOI
高影响因子
Chinese Journal of Natural Medicines
·
2016
·
IF 1.382
Zhuo-Wei SHEN,Meng-Yue LUO,Hai-Hong HU,Hui ZHOU,Hui-Di JIANG,Lu-Shan YU,Su ZENG
NTCP is specifically expressed on the basolateral membrane of hepatocytes, participating in the enterohepatic circulation of bile salts, especially conjugated bile salts, to maintain bile salts homeostasis. In addition, recent studies have found that NTCP is a functional receptor of HBV and HDV. Therefore, it is important to study the interaction between drugs and NTCP and identify the inhibitors/substrates of NTCP. In the present study, a LLC-PK1 cell model stably expressing human NTCP was established, which was simple and suitable for high throughput screening, and utilized to screen and verify the potential inhibitors of NTCP from 102 herbal medicinal ingredients. The results showed that ginkgolic acid (GA) (13 : 0), GA (15 : 1), GA (17 : 1), erythrosine B, silibinin, and emodin have inhibitory effects on NTCP uptake of TCNa in a concentration-dependent manner. Among them, GA (13 : 0) and GA (15 : 1) exhibited the stronger inhibitory effects, with IC 50 values being less than 8.3 and 13.5 μmol·L −1 , respectively, than the classical inhibitor, cyclosporin A (CsA) (IC 50 = 20.33 μmol·L −1 ). Further research demonstrated that GA (13 : 0), GA (15 : 1), GA (17 : 1), silibinin, and emodin were not substrates of NTCP. These findings might contribute to a better understanding of the disposition of the herbal ingredients in vivo , especially in biliary excretion.
研究领域:
药理学
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肝脏病学
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分子生物学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）