Publications by authors named "Yunpeng Fan"

56 Publications

Bioactivity-guided isolation of immunomodulatory compounds from the fruits of Ligustrum lucidum.

J Ethnopharmacol 2021 Jun 30;274:114079. Epub 2021 Mar 30.

College of Veterinary Medicine, Northwest A&F University, 712100, Yangling, PR China. Electronic address:

Ethnopharmacological Relevance: The fruits of Ligustrum lucidum (FLL) W.T. Aiton (Oleaceae) is included in the 2020 "Chinese Pharmacopoeia" and is widely used in traditional Chinese medicine as a tonic. In recent years, FLL has been reported to improve immune function, but the bioactive compounds and mechanisms of FLL remain poorly characterized.

Aim Of The Study: To identify FFL compounds with strong immune activity and explore their molecular mechanisms.

Materials And Methods: The phagocytic activity of RAW264.7 macrophages and proliferation activity of spleen lymphocytes were used to guide the isolation of bioactive compounds from FLL extracts. Lymphocyte subpopulations, Ca concentrations, and surface molecule expression were analyzed using flow cytometry. Cytokine secretion was examined using ELISA. FITC-OVA uptake was observed using fluorescence microscopy. NF-κB activation was analyzed using western blotting.

Results: The extraction and isolation produced ten compounds, namely oleuropeinic acid, nuezhenide, isonuezhenide, salidroside, isoligustrosidic acid, ligulucidumosides A, 8(E)-nuezhenide, hydroxytyrosol, oleuropein, and p-hydroxyphenethyl 7-β-D-glucosideelenolic acid ester were isolated and identified from FLL-Bu-30%. Immunoactivity experiments showed that hydroxytyrosol had the strongest macrophage phagocytotic and lymphocyte proliferation-promoting activities. Further studies showed that hydroxytyrosol could significantly enhance lymphocyte subsets CD3, CD4/CD8, and CD3CD4CD8, promote IL-4, IFN-γ, and TNF-α secretion, and increase intracellular Ca concentrations. In addition, the results from RAW264.7 macrophages showed that hydroxytyrosol increased FITC-OVA uptake, induced TNF-α and IL-1β production, upregulated MHC-II, CD80, and CD86 expression, promoted cytoplasmic IκB-α degradation, and increased nuclear NF-κB p65 levels.

Conclusion: Our study provides substantial evidence regarding the mechanism of the immunomodulatory effects of compounds from FLL.
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http://dx.doi.org/10.1016/j.jep.2021.114079DOI Listing
June 2021

A New Method for Chromosomes Preparation by ATP-Competitive Inhibitor SP600125 Enhancement of Endomitosis in Fish.

Front Bioeng Biotechnol 2020 13;8:606496. Epub 2021 Jan 13.

State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, China.

Previous studies have suggested that 1,9-Pyrazoloanthrone, known as SP600125, can induce cell polyploidization. However, what is the phase of cell cycle arrest caused by SP600125 and the underlying regulation is still an interesting issue to be further addressed. Research in this article shows that SP600125 can block cell cycle progression at the prometaphase of mitosis and cause endomitosis. It is suggested that enhancement of the p53 signaling pathway and weakening of the spindle assembly checkpoint are associated with the SP600125-induced cell cycle arrest. Using preliminary SP600125 treatment, the samples of the cultured fish cells and the fish tissues display a great number of chromosome splitting phases. Summarily, SP600125 can provide a new protocol of chromosomes preparation for karyotype analysis owing to its interference with prometaphase of mitosis.
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http://dx.doi.org/10.3389/fbioe.2020.606496DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7838586PMC
January 2021

Surface plasmon resonance imaging-based biosensor for multiplex and ultrasensitive detection of NSCLC-associated exosomal miRNAs using DNA programmed heterostructure of Au-on-Ag.

Biosens Bioelectron 2021 Mar 22;175:112835. Epub 2020 Nov 22.

Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China. Electronic address:

Exosomal miRNAs are potential tumor biomarkers for early diagnosis of non-small cell lung cancer (NSCLC). Herein, a surface plasmon resonance imaging (SPRi)-based biosensor was developed for simultaneous detection of multiplex NSCLC-associated exosomal miRNAs in a clinical sample using Au-on-Ag heterostructure and DNA tetrahedral framework (DTF). Exosomal miRNAs are captured by various DTF probes immobilized on the gold array chip. Subsequently, single-stranded DNA (ssDNA) functionalized silver nanocube (AgNC) hybridizes with the captured exosomal miRNAs and then the ssDNA-coated Au nanoparticles assembled on the surface of AgNC, forming Au-on-Ag heterostructures as essential labels to realize amplified SPR response. With the aid of DNA programmed Au-on-Ag heterostructure and DTF, the SPRi-based biosensor exhibits wide detection range from 2 fM to 20 nM, ultralow limit of detection of 1.68 fM, enhanced capture efficiency, and improved antifouling capability. Furthermore, the biosensor enables accurate discrimination of NSCLC patients based on detection results of exosomal miRNAs. Overall, this developed biosensor is a promising tool for multiplex exosomal miRNAs detection, providing a new possibility for early diagnosis of NSCLC.
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http://dx.doi.org/10.1016/j.bios.2020.112835DOI Listing
March 2021

Radiosensitizing effects of c‑myc gene knockdown‑induced G2/M phase arrest by intrinsic stimuli via the mitochondrial signaling pathway.

Oncol Rep 2020 Dec 13;44(6):2669-2677. Epub 2020 Oct 13.

Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China.

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents and its long‑term survival rate has stagnated in the past decades. Previous studies have shown that tumors in the G2/M phase are more sensitive to radiotherapy. The proto‑oncogene c‑myc is a transformed member of the myc family and c‑myc‑interacting zinc finger protein‑1 (Miz‑1) is a poly‑Cys2His2 zinc finger (ZF) activator of cell cycle regulator genes, such as the cyclin‑dependent kinase inhibitor p21. C‑myc can repress the expression of p21 by binding to Miz‑1 and abolishing the interaction between Miz‑1 and its co‑activators, which induces G2/M phase arrest. Therefore, the present study investigated the radiosensitizing effects of the c‑myc gene and the sensitizing apoptosis pathway, aiming to identify a more effective combination radiotherapy treatment for osteosarcoma. The present study demonstrated that the c‑myc gene was overexpressed in osteosarcoma cells compared to osteoblasts. Following inhibition of c‑myc gene expression in osteosarcoma cells, tumor proliferation was significantly hindered after inducing G2/M phase arrest via regulating G2/M phase‑associated proteins. Additionally, it was revealed that inhibiting c‑myc gene expression combined with radiotherapy could significantly increase the apoptosis rate of osteosarcoma cells via the mitochondrial signaling pathway. In summary, the present study verified the radiosensitizing effects of c‑myc gene knockdown‑induced G2/M phase arrest, which was achieved by intrinsic stimuli through the mitochondrial signaling pathway.
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http://dx.doi.org/10.3892/or.2020.7806DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7640369PMC
December 2020

Surface plasmon resonance biosensor using hydrogel-AuNP supramolecular spheres for determination of prostate cancer-derived exosomes.

Mikrochim Acta 2020 Oct 6;187(11):590. Epub 2020 Oct 6.

Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.

Based on the hydrogel-AuNP supramolecular sphere (H-Au), a label-free and real-time surface plasmon resonance imaging biosensor has been developed for highly sensitive and specific determination of prostate cancer cell-derived exosomes. After integrating the signal amplification effect of the mass cumulative hydrogel and the LSPR effect of AuNPs with high specific aptamer, the SPRi biosensor for exosome detection exhibited a wide linear range from 1.00 × 10 to 1.00 × 10 particles/mL with a limit of detection of 1.00 × 10 particles/mL. Most importantly, with a strong correlation between the SPRi signal and the t-PSA value measured by the clinical chemiluminescence immunoassay, this biosensor displayed excellent practicability for human serum analysis, which exhibits great potential applications in disease diagnosis and bioanalysis. Prostate cancer has been one of the most threatening diseases in human life and health nowadays. In particular, as cancer metastasizes, it is more likely to cause fracture, paraplegia, and even fatal consequences. However, the predominant t-PSA test needs further improvement for the deficiencies of limited specificity and sensitivity, which is prone to false positive. As one of the noninvasive markers of liquid biopsies, exosome has the potential to be a substitute for t-PSA, which can provide specific and predictive information in disease diagnosis and prognosis. Herein, based on the hydrogel-AuNP supramolecular sphere (H-Au), a label-free and real-time surface plasmon resonance biosensor has been developed for highly sensitive and specific detection of prostate cancer cell-derived exosomes. After integrating the signal amplification effect of mass cumulative hydrogel and LSPR effect of AuNPs with high specific aptamer, this developed SPRi biosensor for exosome detection exhibited a wide linear range from 1.00 × 10 to 1.00 × 10 particles/mL with a limit of detection down to 1.00 × 10 particles/mL. Most importantly, with a strong correlation between the SPRi signal and the t-PSA value measured by the clinical chemiluminescence immunoassay, this biosensor displayed excellent practicability in human serum, which exhibited great potential applications in disease diagnosis and bioanalysis.
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http://dx.doi.org/10.1007/s00604-020-04573-4DOI Listing
October 2020

Expression Profile Analysis of the Cell Cycle in Diploid and Tetraploid red var.

Front Genet 2020 17;11:203. Epub 2020 Mar 17.

State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, China.

Polyploidization often leads to "transcriptome shock," and is considered an important factor in evolution of species. Analysis of the cell cycle, which is associated with survival in polyploidy, has proved useful in investigating polyploidization. Here, we used mRNA sequencing to investigate global expression (in cultured cells) and (in fin and liver tissues) in both the diploid and tetraploid red var.. Differential expression (DE) of genes in diploid (7482, 36.0%) and tetraploid (3787, 18.2%) states suggested that and conditions dramatically change mRNA expression levels. However, of the 20,771 total shared expressed genes, 18,050 (87.0%), including 17,905 (86.2%) non-differentially expressed genes (DEGs) and 145 (0.7%) DEGs between diploids and tetraploids, showed the same expression trends in both cultured cells and liver tissues. Of the DEGs, four of seven genes in the cell cycle pathway had the same expression trends (upregulated in diploids and tetraploids) in both cultured cells and liver tissues. Quantitative PCR analysis confirmed the same expression trends in the nine DEGs associated with regulation of the cell cycle. This research on common characteristics between diploids and tetraploids provides insights into the potential molecular regulatory mechanisms of polyploidization. The steady changes that occur between diploids and tetraploids and show the potential value of studying polyploidy processes using cultured cell lines, especially with respect to cell cycle regulation.
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http://dx.doi.org/10.3389/fgene.2020.00203DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7089929PMC
March 2020

An enzyme-free surface plasmon resonance imaging biosensing method for highly sensitive detection of microRNA based on catalytic hairpin assembly and spherical nucleic acid.

Anal Chim Acta 2020 Apr 27;1108:21-27. Epub 2020 Feb 27.

Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China. Electronic address:

MicroRNAs (miRNAs), considered as therapeutic targets and biomarkers, play important roles in biological processes. Herein, an enzyme-free surface plasmon resonance imaging (SPRi) biosensing method has been developed for miRNA detection based on catalytic hairpin assembly and spherical nucleic acid. The hairpin H1 tethered on the surface of the sensor chip is unfolded by miRNA, and then the hybridized miRNA is released through the displacement of the hairpin H2 for the successive hybridization and assembly process. The emerging DNA fragments on the sensor chip surface after hairpins assembly are further used to hybridize with spherical nucleic acid, inducing a remarkably amplified SPR signal. This biosensing method is highly sensitive to miRNA with a detection limit of 53.7 fM and a linear range of 4 orders of magnitude. Moreover, the biosensor demonstrates good specificity and has the ability to distinguish members of homologous miRNA family even with single base differences. Thus, the SPRi biosensing method may hold a great promise for further application in early clinical diagnosis.
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http://dx.doi.org/10.1016/j.aca.2020.02.055DOI Listing
April 2020

Percutaneous transforaminal endoscopic discectomy for the treatment of lateral recess stenosis secondary occurred the discal fungus infection.

BMC Musculoskelet Disord 2020 Mar 18;21(1):175. Epub 2020 Mar 18.

The affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine Hangzhou, 261# huansha road, Shangcheng District, Hangzhou City, 310006, Zhejiang Province, China.

Background: This is a case of lateral recess stenosis secondary occurred the discal fungus infection treated with percutaneous transforaminal endoscopic discectomy (PTED). There has been no relevant reports before.

Case Presentation: A 49-year-old patient who had taken itraconazole for 13 months for lateral recess stenosis secondary occurred the discal fungus infection complained of gradually worsening radiating pain and numbness in the back and inguinal and inner thigh region of right side. In order to relieve the radiating neuralgia and reduce the damage to spinal stability, the minimally invasive PTED was performed.The patient's prognosis was assessed using Oswestry Disability Index (ODI) and Visual Analogue Scale (VAS). During the follow-up, the patient's ODI and VAS scores were decreased significantly. The radiating pain in the inguinal and inner thigh region of right side were significantly alleviated and the discomfort caused by lower back instability was improved by plaster vest.

Discussion And Conclusion: PTED not only avoids further damage to the stability of the lumbar spine, but also effectively relieves the symptoms of leg neuroradialgia caused by lateral recess stenosis secondary occurred the discal fungus infection.
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http://dx.doi.org/10.1186/s12891-020-03211-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7081584PMC
March 2020

Decompression alone versus fusion and Coflex in the treatment of lumbar degenerative disease: A network meta-analysis.

Medicine (Baltimore) 2020 Mar;99(11):e19457

Department of Orthopedic Surgery, The Affiliated Hangzhou Hospital of Nanjing Medical University.

Background: Lumbar degenerative disease (LDD) is a very common disease. And decompression alone, posterior lumbar interbody fusion (PLIF), and interspinous device (Coflex) are generally accepted surgical techniques. However, the effectiveness and safety of the above techniques are still not clear. Network meta-analysis a comprehensive technique can compare multiple treatments based on indirect dates and all interventions are evaluated and ranked simultaneously. To figure out this problem and offer a better choice for LDD, we performed this network meta-analysis.

Methods: PubMed and WanFang databases were searched based on the following key words, "Coflex," "decompression," "PLIF," "Posterior Lumbar Interbody Fusion," "Coflex" "Lumbar interbody Fusion." Then the studies were sorted out on the basis of inclusion criteria and exclusion criteria. A network meta-analysis was performed using The University of Auckland, Auckland city, New Zealand R 3.5.3 software.

Results: A total of 10 eligible literatures were finally screened, including 946 patients. All studies were randomized controlled trials (RCTs). Compared with decompression alone group, there were no significant differences of Oswestry Disability Index (ODI) in Coflex and lumbar interbody fusion groups after surgery. However, Coflex and PLIF were better in decreasing Visual Analogue Scale (VAS) score compared with decompression alone. Furthermore, we found Coflex have a less complication incidence rate.

Conclusion: Compared with decompression alone, Coflex and lumbar interbody fusion had the similar effectiveness in improving lumbar function and quality of life. However, the latter 2 techniques were better in relieving pain. Furthermore, Coflex included a lower complication incidence rate. So we suggested that Coflex technique was a better choice to cue lumbar spinal stenosis (LSS).

Level Of Evidence: Systematic review and meta-analysis, level I.
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http://dx.doi.org/10.1097/MD.0000000000019457DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7220096PMC
March 2020

High-sensitive and multiplex biosensing assay of NSCLC-derived exosomes via different recognition sites based on SPRi array.

Biosens Bioelectron 2020 Apr 28;154:112066. Epub 2020 Jan 28.

Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China. Electronic address:

Non-small cell lung cancer (NSCLC) have been reported to secret a high concentration of exosomes into blood circulatory system, which is one of sensitive and non-invasive biomarkers for NSCLC's early-stage diagnosis. But it is still lack of feasible and accurate methods to analyze the different NSCLC cells-derived exosomes. Herein, we built a SPRi biosensing assay for high-sensitive and multiplex characterizations of NSCLC-derived exosomes by bioaffinity interactions of antibodies and different recognition sites. By this way, the exosomes derived from normal lung and NSCLC cells can be effectively distinguished through precise identification of the exosomal protein pattern. And the multiplex characterizations of NSCLC-related exosomes are also achieved by anti-CD63, anti-EGFR and anti-EpCAM modified SPRi array. The limit of detection (LOD) of this SPRi-based biosensor approaches to the level of 10 particles/μL with the help of functionalized gold nanoparticles. Besides, the developed biosensing assay was successfully applied in the determination of exosomes purified from clinical plasma samples. This SPRi biosensing strategy might offer a potential alternative for massive high-throughput screening for NSCLC in clinical specimens.
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http://dx.doi.org/10.1016/j.bios.2020.112066DOI Listing
April 2020

Topping-off surgery vs posterior lumbar interbody fusion for degenerative lumbar disease: a finite element analysis.

J Orthop Surg Res 2019 Dec 30;14(1):476. Epub 2019 Dec 30.

Department of Orthopedic Surgery, The Affiliated Hangzhou Hospital of Nanjing Medical University, Hangzhou, 310006, China.

Background: Adjacent segment disease (ASD) is a common complication after posterior lumbar interbody fusion (PLIF). Recently, a topping-off surgery (non-fusion with Coflex) has been developed to reduce the risk of ASD, yet whether and how the topping-off surgery can relieve ASD remains unclear. The purpose of this study was to explore the biomechanical effect of PLIF and Coflex on the adjacent segments via finite element (FE) analysis and discuss the efficacy of Coflex in preventing ASD.

Methods: A FE model of L3-L5 segments was generated based on the CT of a healthy volunteer via three commercially available software. Coflex and PLIF devices were modeled and implanted together with the segment model in the FE software. In the FE model, a pre-compressive load of 500 N, equal to two-thirds of the human body mass, was applied on the top surface of the L3. In addition, four types of moments (anteflexion, rear protraction, bending, and axial rotation) set as 10 Nm were successively applied to the FE model combined with this pre-compressive load. Then, the range of motion (ROM), the torsional rigidity, and the maximum von Mises equivalent stress on the L3-L4 intervertebral disc and the implant were analyzed.

Results: Both Coflex and PLIF reduced ROM. However, no significant difference was found in the maximum von Mises equivalent stress of adjacent segment disc between the two devices. Interestingly enough, both systems increased the torsional rigidity at the adjacent lumbar segment, and PLIF had a more significant increase. The Coflex implant had a larger maximum von Mises equivalent stress.

Conclusions: Both Coflex and PLIF reduced ROM at L3-L4, and thus improved the lumbar stability. Under the same load, both devices had almost the same maximum von Mises equivalent stress as the normal model on the adjacent intervertebral disc. But it is worthy to notice the torsional rigidity of PLIF was higher than that of Coflex, indicating that the lumbar treated with PLIF undertook a larger load to reach ROM of Coflex. Therefore, we presumed that ADS was related to a higher torsional rigidity.
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http://dx.doi.org/10.1186/s13018-019-1503-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6937696PMC
December 2019

Characterization and antioxidant activity of the oligo-maltose fraction from Polygonum Cillinerve.

Carbohydr Polym 2019 Dec 10;226:115307. Epub 2019 Sep 10.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

In this study, Polygonum Cillinerve polysaccharide (PCP) was extracted, and characterized by high performance gel permeation chromatography, high performance liquid chromatography, fourier transform infrared spectroscopy, nuclear magnetic resonance and mass spectroscopy. The results showed that PCP was composed of glucose, it was α-D-glucan and the backbone of PCP was consisted of repeating units of (1→4)-α-D-Glucose. In addition, the antioxidant potential of PCP was assessed in vitro. The results showed that PCP had strong hydroxyl radical scavenging ability, some DPPH scavenging ability and good reducing power. Moreover, the results of the enzyme-linked immunosorbent assays showed that PCP at 15.625-0.975 μg/mL could significantly improve the level of superoxide dismutase and glutathione peroxidase, and could significantly decrease the level of myeloperoxidase, malondialdehyde and xanthine oxidase in macrophages. These results indicated that PCP could potentially be developed as a natural antioxidant.
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http://dx.doi.org/10.1016/j.carbpol.2019.115307DOI Listing
December 2019

Fiber optic surface plasmon resonance biosensor for detection of PDGF-BB in serum based on self-assembled aptamer and antifouling peptide monolayer.

Biosens Bioelectron 2019 Sep 25;140:111350. Epub 2019 May 25.

Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, PR China. Electronic address:

Herein, a home-build fiber optic surface plasmon resonance (FO-SPR) biosensing platform has been developed for highly sensitive detection of platelet-derived growth factor (PDGF-BB) based aptamer-functionalized AuNPs for signal enhancement. In this biosensor, the PDGF-BB aptamer was used to specifically capture PDGF-BB, and the antifouling peptide demonstrated great ability for resisting non-specific adsorption. After a sandwich reaction, the aptamer, PDGF-BB and aptamer-functionalized AuNPs complexes were formed on the fiber optic (FO) probe surface to significantly amplify FO-SPR signal. This method exhibited a broad detection range from 1 to 1000 pM of PDGF-BB and a low detection limit of 0.35 pM. Moreover, this biosensor was successfully applied to the detection of PDGF-BB in 10% human serum samples without suffering from serious interference owing to the excellent antifouling property of the peptide. Thus, this developed FO-SPR biosensor could be a potential alternative device for proteins determination, even as a point-of-care diagnostic tool (POCT) in clinical application.
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http://dx.doi.org/10.1016/j.bios.2019.111350DOI Listing
September 2019

Galectin-1 inhibition induces cell apoptosis through dual suppression of CXCR4 and Ras pathways in human malignant peripheral nerve sheath tumors.

Neuro Oncol 2019 11;21(11):1389-1400

Department of Biochemistry and Molecular Medicine, University of California Davis, Sacramento, California, USA.

Background: The Ras signaling pathway is commonly dysregulated in human malignant peripheral nerve sheath tumors (MPNSTs). It is well known that galectin-1 (Gal-1) is essential to stabilize membrane Ras and thereby induce the activation of Ras. However, the role of Gal-1 in MPNST progression remains unknown. The aim of this study was to examine whether Gal-1 knockdown could have an effect on the Ras signaling pathway.

Methods: Cell viability, apoptosis assay, and colony formation were performed to examine the effects of inhibition of Gal-1 in MPNST cells. We used a human MPNST xenograft model to assess growth and metastasis inhibitory effects of Gal-1 inhibitor LLS2.

Results: Gal-1 was upregulated in MPNST patients and was highly expressed in MPNST cells. Knockdown of Gal-1 by small interfering (si)RNA in Gal-1 expressing MPNST cells significantly reduces cell proliferation through the suppression of C-X-C chemokine receptor type 4 (CXCR4) and the rat sarcoma viral oncogene homolog (RAS)/extracellular signal-regulated kinase (ERK) pathway, which are important oncogenic signaling in MPNST development. Moreover, Gal-1 knockdown induces apoptosis and inhibits colony formation. LLS2, a novel Gal-1 allosteric small molecule inhibitor, is cytotoxic against MPNST cells and was able to induce apoptosis and suppress colony formation in MPNST cells. LLS2 treatment and Gal-1 knockdown exhibited similar effects on the suppression of CXCR4 and RAS/ERK pathways. More importantly, inhibition of Gal-1 expression or function by treatment with either siRNA or LLS2 resulted in significant tumor responses in an MPNST xenograft model.

Conclusion: Our results identified an oncogenic role of Gal-1 in MPNST and that its inhibitor, LLS2, is a potential therapeutic agent, applied topically or systemically, against MPNST.
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http://dx.doi.org/10.1093/neuonc/noz093DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6827838PMC
November 2019

The acaricidal activity and mechanism of eugenol on Psoroptes cuniculi.

Vet Parasitol 2019 Feb 7;266:56-62. Epub 2019 Jan 7.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

In this study, the acaricidal effect of eugenol was measured and its mechanism of action investigated. The results showed that eugenol possessed the effect of killing Psoroptes cuniculi, and could regulate the mRNA expression of glutathione S-transferase (GST), catechinic acid (Ca) and thioredoxin (Trx). PPAR, NF-kappa B, TNF, Rap 1 and Ras signaling pathways might be the main pathways that involved into the process of killing mites. These findings suggested that eugenol could be developed into a new kind of acaricide, and further expand current knowledge on the mechanisms of eugenol for killing Psoroptes cuniculi of eugenol.
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http://dx.doi.org/10.1016/j.vetpar.2018.12.012DOI Listing
February 2019

Phospholipase Cγ2 signalling contributes to the haemostatic effect of Notoginsenoside Ft1.

J Pharm Pharmacol 2019 May 14;71(5):878-886. Epub 2018 Dec 14.

College of Veterinary Medicine, Northwest A&F University, Yangling, China.

Objectives: The drawback of bleeding caused by chronic antiplatelet therapy is persecuting patients with thrombotic diseases severely. Based on the dual-directional regulatory effect of Panax notoginseng on platelet, the present study focused on the effect of Notoginsenoside Ft1, a saponin with effect in promoting platelet aggregation.

Key Findings: According to the present study, Notoginsenoside Ft1 cannot stimulate platelet aggregation independently. However, the effect in enhancing aggregation induced by thrombin, collagen and ADP is peaked at 5-10 μm. In addition, thrombin-induced activation of PLCγ2-IP /DAG-[Ca ]/PKC-TXA signalling was potentiated by Notoginsenoside Ft1, as well. Furthermore, the mice tail bleeding time was shortened by administration of Notoginsenoside Ft1 significantly. And the bleeding time prolonged by aspirin was also restored by Ft1.

Conclusions: The haemostatic effect of Notoginsenoside Ft1 was exerted through potentiation of PLCγ2-IP /DAG-[Ca ]/PKC-TXA signalling pathway stimulated by other stimulators. Notoginsenoside Ft1 has the potential to be developed into supplements in antiplatelet therapy to prevent the drawback of bleeding.
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http://dx.doi.org/10.1111/jphp.13057DOI Listing
May 2019

Phospholipase Cγ2 Signaling Cascade Contribute to the Antiplatelet Effect of Notoginsenoside Fc.

Front Pharmacol 2018 6;9:1293. Epub 2018 Nov 6.

Laboratory of Traditional Chinese Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China.

Bleeding, the main drawback of clinically used chemical anti-thrombotic drug is resulted from the unidirectional suppression of platelet activity. Therefore, dual-directional regulatory effect on platelet is the main preponderance of over these drugs. The dual-directional regulatory effect should be ascribed to the resourceful saponins (PNS). Clarifying the mechanism of main PNS in both inhibiting and promoting platelet aggregation will give a full outlook for the dual-directional regulatory effect. The present study is aimed at explaining the mechanism of Notoginsenoside Fc (Fc), a main PNS, in inhibiting platelet aggregation. In the study, after incubating platelets with Fc and m-3M3FBS, platelet aggregation was triggered by thrombin, collagen or ADP. Platelet aggregation was measured by aggregometer. Phospholipase Cγ2 (PLCγ2) and protein kinase C (PKC) activities were studied by western blotting. Diacylglycerol (DAG), thromboxane B (TXB) and 1,4,5-inositol trisphosphate (IP) concentrations were measured by corresponding ELISA kits. Calcium concentrations ([Ca]) were estimated through the fluorescence intensity emitted from Fluo-4. In the study, thrombus model was induced by FeCl. The effect of Fc on thrombosis was evaluated by measurement of protein content and observation of injured blood vessel. thrombin, collagen and ADP induced platelet aggregation were all suppressed by incubating platelets with Fc. Platelet PLCγ2 and subsequent DAG-PKC-TXA and IP were down-regulated by Fc as well. However, the basal [Ca] in platelet was not altered by Fc. Nevertheless, thrombin triggered activation of PLCγ2 and subsequent DAG-PKC-TXA and IP-[Ca] were all abolished by Fc. Fc also attenuated platelet aggregation and PLCγ2 signaling activation induced by PLC activator, m-3M3FBS. In the study, FeCl induced thrombosis in rat femoral artery was significantly alleviated by administration of Fc. The results above suggested the antiplatelet and antithrombotic effects of Fc are carried out through oppression of PLCγ2 and subsequent DAG-PKC-TXA and IP-[Ca]. The present study provided theoretical support for new anti-thrombotic drug exploitation by .
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http://dx.doi.org/10.3389/fphar.2018.01293DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6232503PMC
November 2018

Enhanced Resistance of Triploid Crucian Carp to Cadmiuminduced Oxidative and Endoplasmic Reticulum Stresses.

Curr Mol Med 2018 ;18(6):400-408

State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, Hunan, China.

Background: Cadmium is a major heavy metal pollutant. Even at low concentrations in waste water Cadmium can accumulate in algae and sediments, and it is absorbed by both plants and aquatic animals, like shellfish and fish. Triploid crucian carp is an important economic fish, and have been farmed on a large scale in China.

Objective: to explore the molecular mechanisms underlying Cadmium stress of triploid crucian carp.

Method: we applied following experimental method to conduct research: transcriptomes analysis, histological observation, Quantitative real-time PCR and enzyme activity analysis et al. Results: in the present study, we obtained following important results: 1) Under Cd stress, the mortality and abnormality rates in triploid crucian carp were lower than those of diploid strains and the effects of Cadmium treatment on the liver were revealed by histological observation; 2) the liver transcriptomes of triploid crucian carp were obtained and the data was analyzed; 3) 14 genes related to algae and sediments stress were screened from transcriptome data, and the expression of these genes was verified by Quantitative real-time PCR. The results were consistent with the gene results in transcriptome data; 4) Quantitative real-time PCR and enzyme activity analysis all confirmed triploid crucian carp resistance to Cadmium stress was regulated by both oxidative stress and ER stress responses; 5) We found that IRE-1 and PERK, not ATF- 6, were involved in the enhance Cadmium resistance of triploid crucian carp.

Conclusion: these studies demonstrate that triploid crucian carp have strong resistance to Cadmium.
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http://dx.doi.org/10.2174/1566524018666181113105018DOI Listing
September 2019

Acute immune stress improves cell resistance to chemical poison damage in SP600125-induced polyploidy of fish cells in vitro.

Fish Shellfish Immunol 2019 Jan 25;84:656-663. Epub 2018 Oct 25.

State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, Hunan, 410081, PR China; School of Life Sciences, Hunan Normal University, Changsha, Hunan, 410081, PR China. Electronic address:

Previous research has indicated that the small compound, SP600125, could induce polyploidy of fish cells, and has established a stable tetraploid cell line from diploid fish cells. In order to explore how fish cells maintain homeostasis under SP600125-stress in vitro, this study investigates impacts of SP600125-stress on intracellular pathways, as well as on regulation of the cellular homeostasis feedback in fish cells. Transcriptomes are obtained from the SP600125-treated cells. Compared with unigenes expressed in control group (crucial carp fin cells), a total of 2670 and 1846 unigenes are significantly upregulated and downregulated in these cells, respectively. Differentially expressed genes are found, which are involved in innate defense, inflammatory pathways and cell adhesion molecules-related pathways. The SP600125-stress enhances cell-mediated immunity, characterized by significantly increasing expression of multiple immune genes. These enhanced immune genes include the pro-inflammatory cytokines (IL-1β, TNF-ɑ, IL-6R), the adaptor signal transducers (STAT, IκBɑ), and the integrins (ɑ2β1, ɑMβ2). Furthermore, mitochondria are contributed to the cellular homeostasis regulation upon the SP600125-stress. The results show that acute inflammation is an adaptive and controlled response to the SP600125-stress, which is beneficial for alleviating toxicity by SP600125. They provide a potential way of breeding fish polyploidy induced by SP600125 in the future research.
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http://dx.doi.org/10.1016/j.fsi.2018.10.063DOI Listing
January 2019

The immunoregulatory activities of astragalus polysaccharide liposome on macrophages and dendritic cells.

Int J Biol Macromol 2017 Dec 19;105(Pt 1):852-861. Epub 2017 Jul 19.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

The objective of this study is to investigate the immunomodulatory activities of astragalus polysaccharide liposome (APSL) on murine peritoneal macrophages and bone marrow derived dendritic cells (DCs). The results showed that APSL not only significantly improved the phagocytosis of macrophages, the contents of IL-6 and IL-12, and the secretion of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) in macrophages, but also promoted the proliferation of DCs precursor cells, enhanced the abilities of DCs on stimulating T-cell proliferation and presenting antigen, upregulated IFN-γ and IL-2 production of DCs, and improved the expression of CD80 and CD86 in DCs compared with astragalus polysaccharide (APS). These findings indicated that the immune-modulating activities of APS were remarkably enhanced after encapsulated with liposome, and liposome possessed the potential to act as effective drug delivery system. Moreover, it also provided the theoretical basis for further researching the mechanism of APSL on improving the immune response.
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http://dx.doi.org/10.1016/j.ijbiomac.2017.07.108DOI Listing
December 2017

Licocoumarone isolated from Glycyrrhiza uralensis selectively alters LPS-induced inflammatory responses in RAW 264.7 macrophages.

Eur J Pharmacol 2017 Apr 2;801:46-53. Epub 2017 Mar 2.

School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; Institute of Chinese Medical Sciences, University of Macau, Macau, SAR, China. Electronic address:

The effects of licocoumarone (LC) isolated from Glycyrrhiza uralensis were studied in LPS-stimulated RAW 264.7 macrophages. Our study demonstrated that LC dose-dependently attenuated LPS-induced NO production by down-regulating iNOS expression. Additionally, the treatment with LC inhibited LPS-induced expression of cytokines including IL-1β, IL-6 and IL-10, but not TNF-α, at both mRNA and protein levels. Similar suppressive effects of LC were observed on LPS-stimulated murine peritoneal macrophages as well. Furthermore, LC significantly reduced LPS-stimulated NF-κB activation by inhibition of IκBα degradation and p65 phosphorylation. The results from NF-κB-luc reporter gene assay further support the inhibitory effect of LC on NF-κB activation. Further studies showed that LC also interfered with the MAPKs and STAT3 signaling pathways, which are typical inflammatory signaling pathways triggered by LPS. Taken together, these results show that LC attenuates LPS-induced cytokine gene expression in RAW 264.7 macrophages through mechanisms that involve NF-κB, MAPKs and STAT3 signaling pathways, but the pattern of inhibition differs from that of a global immunosuppresant. Our study indicates that LC is a functional constituent of Glycyrrhiza uralensis with potential implications in infectious and immune-related diseases.
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http://dx.doi.org/10.1016/j.ejphar.2017.02.049DOI Listing
April 2017

Activation of macrophages by the ophiopogon polysaccharide liposome from the root tuber of Ophiopogon japonicus.

Int J Biol Macromol 2016 Oct 14;91:918-25. Epub 2016 Jun 14.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

This study evaluated the immunomodulatory effects of ophiopogon polysaccharide liposome (OPL) on macrophages in vitro. The phagocytic activity, the secretion of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), the level of cytokines, chemokines and the expression of CD14 and MHC-II costimulatory molecules were measured. Results showed that OPL could significantly improve the phagocytic activity and the level of IL-1β, TNF-α, MCP-1 and MIP-1β, promote the secretion of NO and iNOS, and enhance the expression of CD14 and MHC-II costimulatory molecules in the peritoneal macrophages of mice compared with ophiopogon polysaccharide (OP). Altogether, these results suggested that OPL could activate macrophages, and the efficacy was significantly superior to OP. Therefore, OPL would be exploited in a potent immunomodulators. Moreover, it also provided the theoretical basis for further studying the mechanism of OPL on improving the immune response.
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http://dx.doi.org/10.1016/j.ijbiomac.2016.06.037DOI Listing
October 2016

Purification of flavonoids from licorice using an off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method.

J Sep Sci 2016 Jul 17;39(14):2710-9. Epub 2016 Jun 17.

Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai, P. R. China.

An orthogonal (71.9%) off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method coupled with effective sample pretreatment was developed for separation and purification of flavonoids from licorice. Most of the nonflavonoids were firstly removed using a self-made Click TE-Cys (60 μm) solid-phase extraction. In the first dimension, an industrial grade preparative chromatography was employed to purify the crude flavonoids. Click TE-Cys (10 μm) was selected as the stationary phase that provided an excellent separation with high reproducibility. Ethyl acetate/ethanol was selected as the mobile phase owing to their excellent solubility for flavonoids. Flavonoids co-eluted in the first dimension were selected for further purification using reversed-phase liquid chromatography. Multiple compounds could be isolated from one normal-phase fraction and some compounds with bad resolution in one-dimensional liquid chromatography could be prepared in this two-dimensional system owing to the orthogonal separation. Moreover, this two-dimensional liquid chromatography method was beneficial for the preparation of relatively trace flavonoid compounds, which were enriched in the first dimension and further purified in the second dimension. Totally, 24 flavonoid compounds with high purity were obtained. The results demonstrated that the off-line two-dimensional liquid chromatography method was effective for the preparative separation and purification of flavonoids from licorice.
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http://dx.doi.org/10.1002/jssc.201501393DOI Listing
July 2016

The anti-tumor activity and mechanism of alkaloids from Aconitum szechenyianum Gay.

Bioorg Med Chem Lett 2016 Jan 10;26(2):380-387. Epub 2015 Dec 10.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

In the current study, the anti-tumor activity of Aconitum szechenyianum Gay alkaloids (ASA) and its mechanism of action were investigated. The result showed that ASA could induce apoptosis in HepG-2, Hela and A549 cells but not in normal human embryonic kidney 293A cells, and its apoptotic effect on A549 cells was stronger than those of HepG-2 and Hela cells. Moreover, the following study showed that ASA could up-regulate the expression levels of p38 and phosphorylated p38 MAPK, suggesting ASA-induced apoptosis was associated with the p38 MAPK mediated pathway. Furthermore, ASA could up-regulate TNF-R1 and DR5 via activation of p38 MAPK, thereby activating caspase 8, revealing the death receptor pathway was also involved in this process. In addition, ASA could led to a loss in the mitochondrial out membrane potential, up-regulate p53, phosphorylated p53 and Bax, down-regulate Bcl-2, release cytochrome c from the mitochondria to the cytoplasm, and activate caspase-9 and caspase-3 in A549 cells, which revealed that ASA could also induce apoptosis through the mitochondria mediated pathway. These results suggested that ASA played the anti-tumor role through the activation of p38 MAPK-, death receptor-, mitochondria- and caspase-dependent apoptotic pathways.
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http://dx.doi.org/10.1016/j.bmcl.2015.12.006DOI Listing
January 2016

The Adjuvant Activity of Epimedium Polysaccharide-Propolis Flavone Liposome on Enhancing Immune Responses to Inactivated Porcine Circovirus Vaccine in Mice.

Evid Based Complement Alternat Med 2015 3;2015:972083. Epub 2015 Nov 3.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.

Objectives. The adjuvant activity of Epimedium polysaccharide-propolis flavone liposome (EPL) was investigated in vitro and in vivo. Methods. In vitro, the effects of EPL at different concentrations on splenic lymphocytes proliferation and mRNA expression of IFN-γ and IL-6 were determined. In vivo, the adjuvant activities of EPL, EP, and mineral oil were compared in BALB/c mice through vaccination with inactivated porcine circovirus type 2 (PCV2) vaccine. Results. In vitro, EPL promoted lymphocytes proliferation and increased the mRNA expression of IFN-γ and IL-6, and the effect was significantly better than EP at all concentrations. In vivo, EPL significantly promoted the lymphocytes proliferation and the secretion of cytokines and improved the killing activity of NK cells, PCV2-specific antibody titers, and the proportion of T-cell subgroups. The effects of EPL were significantly better than EP and oil adjuvant at most time points. Conclusion. EPL could significantly improve both PCV2-specific cellular and humoral immune responses, and its medium dose had the best efficacy. Therefore, EPL would be exploited in an effective immune adjuvant for inactivated PCV2 vaccine.
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http://dx.doi.org/10.1155/2015/972083DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4647051PMC
November 2015

The adjuvanticity of ophiopogon polysaccharide liposome against an inactivated porcine parvovirus vaccine in mice.

Int J Biol Macromol 2016 Jan 31;82:264-72. Epub 2015 Oct 31.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

In this study, the adjuvant activity of ophiopogon polysaccharide liposome (OPL) was investigated. The effects of OPL on the splenic lymphocyte proliferation of mice were measured in vitro. The results showed that OPL could significantly promote lymphocyte proliferation singly or synergistically with PHA and LPS and that the effect was better than ophiopogon polysaccharide (OP) at most of concentrations. The adjuvant activities of OPL, OP and mineral oil were compared in BALB/c mice inoculated with inactivated PPV in vivo. The results showed that OPL could significantly enhance lymphocyte proliferation, increase the proportion of CD4(+) and CD8(+) T cells, improve the HI antibody titre and specific IgG response, and promote the production of cytokines, and the efficacy of OPL was significantly better than that of OP. In addition, OPL significantly improved the cellular immune response compared with oil adjuvant. These results suggested that OPL possess superior adjuvanticity and that a medium dose had the best efficacy. Therefore, OPL can be used as an effective immune adjuvant for an inactivated PPV vaccine.
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http://dx.doi.org/10.1016/j.ijbiomac.2015.10.084DOI Listing
January 2016

Antioxidative and immunological activities of ophiopogon polysaccharide liposome from the root of Ophiopogon japonicus.

Carbohydr Polym 2016 Jan 31;135:110-20. Epub 2015 Aug 31.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

The antioxidative and immunoregulatory activities of ophiopogon polysaccharide liposome (OPL) were investigated. The results demonstrated that the radical-scavenging activity increased along with the increasing of OPL concentrations, and the scavenging capability of OPL was slightly stronger than ophiopogon polysaccharide (OP) in vitro. In vivo, OPL at high and medium doses could significantly improve the levels of SOD and GSH-Px, and decrease the levels of MPO, XOD and MDA compared with OP. In addition, OPL could not only significantly enhance the splenocyte proliferation and the proportion of CD4(+) and CD8(+) T cells, but also improve the levels of cytokines, antigen-specific antibody titers and immune organ index in the mice immunized with OVA compared with OP. These results indicated that the antioxidative and immunoregulatory activities of OP were significantly enhanced after encapsulated with liposome. Therefore, OPL would be expected to exploit into a new-type of functional food as both medicine and food.
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http://dx.doi.org/10.1016/j.carbpol.2015.08.089DOI Listing
January 2016

The activation of Epimedium polysaccharide-propolis flavone liposome on Kupffer cells.

Carbohydr Polym 2015 Nov 17;133:613-23. Epub 2015 Jul 17.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China. Electronic address:

Epimedium polysaccharide-propolis flavone liposome (EPL), a potent immunological pharmaceutical preparation, was investigated for the immunomodulatory activity on Kupffer cells (KCs) in vitro. The results showed that EPL could significantly induce the secretion of chemokines (RANTES and MCP-1), promote the production of nitric oxide and induced nitric oxide synthase, improve the pinocytic and phagocytic activity of KCs, promote the mRNA expression of TNF-α and IL-1β, and enhance the expression of costimulatory molecules (CD11b and CD68) in KCs compared with Epimedium polysaccharide-propolis flavone (EP) at 30-7.5μg/mL. In addition, the abilities of KCs on stimulating lymphocytes proliferation and antigen presenting were significantly enhanced after stimulated with EPL compared with EP. These results suggested that EPL could activate KCs and possessed the stronger immunomodulatory effect, which provided the theoretical basis for further studying the mechanism of EPL on improving the immune response.
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http://dx.doi.org/10.1016/j.carbpol.2015.07.044DOI Listing
November 2015

The anti-porcine parvovirus activity of nanometer propolis flavone and propolis flavone in vitro and in vivo.

Evid Based Complement Alternat Med 2015 26;2015:472876. Epub 2015 Feb 26.

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.

Objectives. The present study was conducted to evaluate the activity of nanometer propolis flavone (NPF) on inhibiting porcine parvovirus (PPV) in vitro and in vivo. Methods. In vitro, the effect of NPF on cellular infectivity of PPV was carried out before and after adding drug and simultaneous adding and PPV after being mixed. In vivo, the anti-PPV effect of NPF in guinea pigs was performed. Results. The results showed that NPF could significantly inhibit PPV infecting porcine kidney- (PK-) 15 cells compared with propolis flavone (PF), and the activity of NPF was the best in preadding drug pattern. NPF at high and medium doses was able to observably restrain PPV copying in lung, gonad, blood, and spleen, decrease the impact of PPV on weight of guinea pigs, and improve hemagglutination inhibition (HI) of PPV in serum. In addition, it could also increase the contents of IL-2 and IL-6 in serum after PPV challenge. Conclusion. These results indicated that NPF could significantly improve the anti-PPV activity of PF, and its high concentration possessed the best efficacy. Therefore, NPF would be expected to be exploited into a new-style antiviral drug.
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http://dx.doi.org/10.1155/2015/472876DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357139PMC
March 2015

Qualitative and quantitative analysis of an alkaloid fraction from Piper longum L. using ultra-high performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry.

J Pharm Biomed Anal 2015 May 16;109:28-35. Epub 2015 Feb 16.

Engineering Research Center of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, PR China. Electronic address:

In a previous research, an alkaloid fraction and 18 alkaloid compounds were prepared from Piper longum L. by series of purification process. In this paper, a qualitative and quantitative analysis method using ultra-high performance liquid chromatography-diode array detector-mass spectrometry (UHPLC-DAD-MS) was developed to evaluate the alkaloid fraction. Qualitative analysis of the alkaloid fraction was firstly completed by UHPLC-DAD method and 18 amide alkaloid compounds were identified. A further qualitative analysis of the alkaloid fraction was accomplished by UHPLC-MS/MS method. Another 25 amide alkaloids were identified according to their characteristic ions and neutral losses. At last, a quantitative method for the alkaloid fraction was established using four marker compounds including piperine, pipernonatine, guineensine and N-isobutyl-2E,4E-octadecadienamide. After the validation of this method, the contents of above four marker compounds in the alkaloid fraction were 57.5mg/g, 65.6mg/g, 17.7mg/g and 23.9mg/g, respectively. Moreover, the relative response factors of other three compounds to piperine were calculated. A comparative study between external standard quantification and relative response factor quantification proved no remarkable difference. UHPLC-DAD-MS method was demonstrated to be a powerful tool for the characterization of the alkaloid fraction from P. longum L. and the result proved that the quality of alkaloid fraction was efficiently improved after appropriate purification.
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http://dx.doi.org/10.1016/j.jpba.2015.02.012DOI Listing
May 2015