Fan Luo

Wuhan University, Wuhan, Hubei, China

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Publications (14)21.55 Total impact

  • Article: Establishment of SYBR green-based qPCR assay for rapid evaluation and quantification for anti-Hantaan virus compounds in vitro and in suckling mice.
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    ABSTRACT: Hantaan viruses cause two severe diseases lacking efficient treatment, yet no effective prophylactic vaccines are available. Continued exploration of alternative antiviral agents to treat hantavirus-related syndromes remains compulsory. The fluorescence-based quantitative real-time PCR (qPCR) has become the touchstone for target gene quantification. In the present study, standard curves for Hantaan virus (HTNV), mouse, and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were generated by serial 10-fold dilutions of the constructed recombinant plasmid pGEM-T/HTNV, pGEM-T/mouse-GAPDH, and pGEM-T/human-GAPDH, respectively. Comparisons between the indirect immunofluorescence assay and qPCR assay in the detection of HTNV-infected Vero E6 cells showed improved detection limit and sensitivity of latter method. To characterize the inhibitory effect of several conventional antivirals (arbidol and ribavirin) and unconventional antivirals (indomethacin and curcumin) on HTNV, the levels of viral RNAs were measured for 4 days post-treatment of HTNV-infected Vero E6 cells and 18 days post-inoculation of HTNV-infected suckling mice. Our results validated that HTNV was sensitive to ribavirin and arbidol treatment, while indomethacin and curcumin may also be therapeutically effective in treating HTNV infection. As a result, the establishment and application of qPCR may be a useful tool for the evaluation of potential antivirals for Hantaan virus infection in vitro and in vivo.
    Virus Genes 10/2012; · 1.85 Impact Factor
  • Article: Amelioration of influenza virus-induced reactive oxygen species formation by epigallocatechin gallate derived from green tea.
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    ABSTRACT: Aim:To study whether epigallocatechin gallate (EGCG), a green tea-derived polyphenol, exerted anti-influenza A virus activity in vitro and in vivo.Methods:Madin-Darby canine kidney (MDCK) cells were tested. The antiviral activity of EGCG in the cells was determined using hemagglutination assay and qPCR. Time of addition assay was performed to determine the kinetics of inhibition of influenza A by EGCG. The level of reactive oxygen species (ROS) were determined with confocal microscopy and flow cytometry. BALB/c mice were treated with EGCG (10, 20 or 40 mg·kg(-1)·d(-1), po) for 5 d. On the 3rd d of the treatment, the mice were infected with influenza A virus. Histopathological changes, lung index and virus titers in the lungs were determined.Results:Treatment of influenza A-infected MDCK cells with EGCG (1.25-100 nmol/L) inhibited influenza A replication in a concentration-dependent manner (the ED(50) value was 8.71±1.11 nmol/L). Treatment with EGCG (20 nmol/L) significantly suppressed the increased ROS level in MDCK cells following influenza A infection. In BALB/c mice infected with influenza virus, oral administration of EGCG (40 mg·kg(-1)·d(-1)) dramatically improved the survival rate, decreased the mean virus yields and mitigated viral pneumonia in the lungs, which was equivalent to oral administration of oseltamivir (40 mg·kg(-1)·d(-1)), a positive control drug.Conclusion:The results provide a molecular basis for development of EGCG as a novel and safe chemopreventive agent for influenza A infection.
    Acta Pharmacologica Sinica 09/2012; · 1.95 Impact Factor
  • Article: [Genetic analysis on S segment of hantaviruses in rodent hosts from Wuhan area, Hubei province].
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    ABSTRACT: To investigate the infection and genotype of hantaviruses in rodents from Wuhan area, Hubei province. Rodents were trapped in fields and residential areas of Xinzhou and Jiangxia districts of Wuhan in autumn and winter seasons, from 2000 to 2003 and from 2009 to 2011. Trapped rodents were identified, and hantavirus antigens were detected in the lung tissues with indirect immunofluorescence assay (IFA). Partial S segment sequences were amplified with RT-PCR in hantavirus antigen positive samples and then sequenced. Phylogenetic tree was constructed to analyze the genetic characteristics of hantaviruses. From 2000 to 2003, 437 rodents were trapped, with 24 (5.49%) lung tissues showed hantavirus antigen positive. From 2009 to 2011, 173 rodents were trapped and 7 (4.05%) were hantavirus antigen positive. Rattus norvegicus were the dominant species of rodents. Partial S segment sequences were amplified from 22 samples with Hantaan and Seoul viruses specific primers and sequenced. Partial S segments of Seoul viruses (nucleotide 588 - 1147) were amplified from 17 rodents (13 R. norvegicus and 4 Apodemus agrarius). Seven of these sequences belonged to 3 genetic lineage, while two novel genetic lineages were formed by 9 and 1 sequences, respectively. Partial S segments of Hantaan viruses (nucleotide 615 - 1141) were amplified from 5 A. agrarius. One of these sequences belonged to 7 genetic lineages, and 4 sequences formed one novel genetic subtype. Hantaan and Seoul viruses co-circulated in Wuhan area, Hubei province. Novel genetic lineages were identified in this study and Seoul virus might have caused spillover infection in A. agrarius.
    Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 08/2012; 33(8):828-31.
  • Article: Genetic characterization of a new subtype of Hantaan virus isolated from a hemorrhagic fever with renal syndrome (HFRS) epidemic area in Hubei Province, China.
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    ABSTRACT: To characterize hantaviruses currently circulating in the hemorrhagic fever with renal syndrome (HFRS) epidemic area of Hubei Province, rodents were captured and serum samples were collected from several HFRS patients. The partial S segment of the hantaviruses amplified from two serum samples had a high degree of sequence identity to the corresponding hantavirus strain isolated from Apodemus agrarius (designated as HV004). The complete S, M, and L segment sequences of HV004 were determined. The sequence identities between strain HV004 and other Hantaan viruses (HTNVs) were 83 %-90 % at the nucleotide level and 95 %-99 % at the amino acid level. Phylogenetic analysis showed that HV004 belonged to a new HTNV lineage. These data suggest the presence of a new HTNV subtype, which probably caused the HFRS cases in the endemic area of Hubei Province.
    Archives of Virology 06/2012; 157(10):1981-7. · 2.11 Impact Factor
  • Article: The inhibitory effect of Rheum palmatum against coxsackievirus B3 in vitro and in vivo.
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    ABSTRACT: Coxsackievirus B(3)(CVB(3)) infection is the major cause of viral myocarditis, as well as dilated cardiomypathy. Rhubarb is one of the oldest and best-known traditional Chinese medicines. We initiated this study to determine the antiviral effect of an ethanol extract from the roots and rhizoma of Rheum palmatum (R. palmatum, one of the Chinese Rhubarbs), against CVB(3) in tissue culture cells and in a mouse model. The ethanol extract from R. palmatum showed significant inhibitory activity against CVB(3) on HEp-2 cells when added after infection, with IC(50) of 4 μg/ml, TI of 10. The medicated mouse serum still contained the pharmaceutical compound 24 h after intraperitoneal injection, and exhibited an antiviral effect on CVB(3)-infected cells, especially in the 0.3 and 0.5 g/kg/day treatment groups. Furthermore, the CVB(3)-infected mice were treated with the extract solution with dosages of 0.3 g/kg/day beginning 24 h post-CVB(3) exposures. The ethanol extract treated mice showed alleviated clinical signs, better survival rate, prolonged MTD and decreased viral titers compared to the virus control group. Our results indicate that the ethanol extract from R. palmatum has the anti-CVB(3) activity in vitro and in vivo and thus provides a re-evaluation of this old remedy with a broad therapeutic potential.
    The American Journal of Chinese Medicine 01/2012; 40(4):801-12. · 1.98 Impact Factor
  • Article: Genetic analysis of hantaviruses and their rodent hosts in central-south China.
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    ABSTRACT: Hantaan virus (HTNV) and Seoul virus (SEOV) are two major zoonotic pathogens of hemorrhagic fever with renal syndrome (HFRS) in Asia. Hubei province, which is located in the central-south China, had been one of the most severe epidemic areas of HFRS. To investigate phylogenetic relationships, genetic diversity and geographic distribution of HTNV and SEOV in their reservoir hosts, a total of 687 rodents were trapped in this area between 2000 and 2009. Sequences of partial S- and M-segments of hantaviruses and mitochondrial D-loop gene from 30 positive samples were determined. Our data indicated that SEOV and HTNV were co-circulating in Hubei. Phylogenetic analysis based on partial S- and M-segment sequences revealed two and three previously undefined lineages of SEOV, and a novel genetic lineage of HTNV, respectively. Four inter-lineage reassortment SEOVs carried by Rattus norvegicus and Apodemus agrarius were observed. It suggests that SEOV may cause spillover infections to A. agrarius naturally. The abundance of the phylogenetic lineages of SEOV suggested that central-south China was a radiation center for SEOVs.
    Virus Research 11/2011; 163(2):439-47. · 2.94 Impact Factor
  • Article: Wavelet-Based Image Registration and Segmentation Framework for the Quantitative Evaluation of Hydrocephalus.
    Int. J. Biomedical Imaging. 01/2010; 2010.
  • Article: [Association of hemorrhagic fever with renal syndrome and HLA-DRB allele polymorphisms in Han nationality, Hubei province].
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    ABSTRACT: To understand the association of human leukocyte antigen (HLA)-DRB polymorphism and patients diagnosed as hemorrhagic fever with renal syndrome (HFRS). HLA-DR allele polymorphism was detected by PCR-sequence specific primers (PCR-SSP). Hantavirus (HV) typed as Hantaan virus (HTNV) and Seoul virus (SEOV) in patients were detected by RT-heminested PCR. The gene frequency of DRB1*0401-0411, *1001 and *1101-1105 in HFRS case group were 3.1%, 2.2% and 15.7% respectively. Compared with control group, it was significant higher in HFRS case group (RR = 13.87, 9.72 and 2.00 respectively with Chi-square value as 10.006, 6.324 and 6.472 respectively, P < 0.05). When compared with HFRS case group, the gene frequency of DRB1*1501-1502, DRB4 and DRB5 in control group were 11.0%, 19.0% and 16.9% respectively, markedly lower than in patients (RR = 0.45, 0.58 and 0.23 respectively. Chi-square values were 6.138, 4.583 and 21.076 respectively, P < 0.05). There was no significant difference in other HLA-DR gene frequencies. Mixed infection was found in Hubei, with HTNV slightly more than SEOV. Distinct hantaviruses could coexist in either different or the same geographic or ecological zores in Hubei province. Patients with HLA-DRB1*1101-1105 alleles were 81.8% (27/33) infected by HTNV and only 18.2% infected by SEOV, which had significant difference (P < 0.05). DRB1*0401-0411, *1001 and *1101-1105 were possibly associated with increased susceptibility to HV infection. On the other hand there was an inverse correlation among HFRS, DRB1*1501-1502, DRB4 and DRB5.
    Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 11/2009; 30(11):1143-6.
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    Article: Efficacy of arbidol on lethal hantaan virus infections in suckling mice and in vitro.
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    ABSTRACT: Arbidol is an immunomodulator that was first developed in Russia. In this study, we report the antiviral activity of arbidol against Hantaan virus (HTNV) in vitro and in vivo. The antiviral activity of arbidol in vitro was determined by plaque-forming assay, ranging from 0.5 to 8 microg/mL. To investigate whether arbidol has an antiviral effect in vivo, suckling BALB/c mice infected with HTNV were treated with arbidol at 24 h before infection with a 5, 10 or 20 mg.kg(-1).d(-1), once per day, for 10 days. On day 12 and 28 post infection (pi), histopathological changes and viral antigen were detected. On days 4, 8, 12, and 16 pi, the viral load of target organs and serum TNF-alpha levels of arbidol-treated animals were determined. Arbidol was found to have potent inhibitory activity against HTNV when added in vitro before or after viral infection, with a 50% inhibitory concentration (IC(50)) of 0.9 and 1.2 microg/mL, respectively. The 50% lethal dose (LD(50)) of arbidol for suckling mice was 78.42 mg.kg(-1).d(-1). Oral administration of arbidol increased both survival rate and mean time to death (MTD). Treatment with arbidol reduced histopathological changes, decreased viral load and viral antigen levels, and modulated the level of serum TNF-alpha. Arbidol has the ability to elicit protective antiviral activity against HTNV in vivo and in vitro.Acta Pharmacologica Sinica (2009) 30: 1015-1024; doi: 10.1038/aps.2009.53; published online 8 June 2009.
    Acta Pharmacologica Sinica 08/2009; 30(7):1015-24. · 1.95 Impact Factor
  • Article: Efficacy of arbidol on lethal hantaan virus infections in suckling mice and in vitro
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    ABSTRACT: Aim: Arbidol is an immunomodulator that was first developed in Russia. In this study, we report the antiviral activity of arbidol against Hantaan virus (HTNV) in vitro and in vivo.
    Acta Pharmacologica Sinica 06/2009; 30(7):1015-1024. · 1.95 Impact Factor
  • Article: Preparation by low-temperature nonthermal plasma of graphite fiber and its characteristics for solid-phase microextraction.
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    ABSTRACT: Low-temperature nonthermal plasma has been used to prepare solid-phase microextraction (SPME) fibers with high adsorbability, long-term serviceability, and high reproducibility. Graphite rods serving as fiber precursors were treated by an air plasma discharged at 15.2-15.5 kV for a duration of 8 min. Sampling results revealed that the adsorptive capacity of the homemade fiber was 2.5-34.6 times that of a polyacrylate (PA) fiber for alcohols (methanol, ethanol, isopropyl alcohol, n-butyl alcohol), and about 1.4-1.6 times and 2.5-5.1 times that of an activated carbon fiber (ACF) for alcohols and BTEX (benzene, toluene, ethylbenzene, and xylenes), respectively. It is confirmed from FTIR (Fourier transform infrared spectrophotometer) and SEM (scanning electron microscope) analyses that the improvement in the adsorptive performance attributed to increased surface energy and roughness of the graphite fiber. Using gas chromatography (GC)-flame-ionization detector (FID), the limits of detection (LODs) of the alcohols and BTEX ranged between 0.19 and 3.75 microg L(-1), the linear ranges were between 0.6 and 35,619 microg L(-1) with good linearity (R(2)=0.9964-0.9997). It was demonstrated that nonthermal plasma offers a fast and simple method for preparing an efficient graphite SPME fiber, and that SPME using the homemade fiber represents a sensitive and selective extraction method for the analysis of a wide range of organic compounds.
    Analytica chimica acta 02/2009; 631(1):62-8. · 4.31 Impact Factor
  • Conference Proceeding: Applications of Electrochemical Technique for Organic Wastewater Treatment
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    ABSTRACT: This paper summarized the application of electrochemical technique for wastewater treatment, including wastewater disinfection and degradation of organic pollutants, especially for toxic and biorefractory organic pollutants, such as phenolic pollutants.
    Bioinformatics and Biomedical Engineering, 2008. ICBBE 2008. The 2nd International Conference on; 06/2008
  • Article: [Study on supramolecular complexes of cyclodextrins and fenoxaprop-p-ethyl by UV spectra].
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    ABSTRACT: The supramolecular complexes of fenoxaprop-p-ethyl and various cyclodextrins (beta-CD, HP-beta-CD, RAMEB) were studied by UV spectra technique. The results showed that all cyclodextrins could form 1:1 inclusion complexes with fenoxaprop-p-ethyl in ethanol-water (the volume ratio of ethanol to water was 1 : 4), and the binding constants of the three types of cyclodextrins toward fenoxaprop-p-ethyl were in the order of KRAEB>KHP-beta-CD>Kbeta-CD. Between 298. 15 and 318. 15 K, the binding constants decreased with temperature increasing. The Gibbs free energy deltaG, enthalpy change deltaH, and entropy change deltaS determined were all negative, suggesting that the inclusion complexation is exothermic and can spontaneously occur by the balance of enthalpy driving and entropy opposing.
    Guang pu xue yu guang pu fen xi = Guang pu 01/2007; 26(12):2298-301. · 0.84 Impact Factor
  • Article: Effects of cyclodextrins on hydrolysis of malathion.
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    ABSTRACT: Cyclodextrins (CDs), with hydrophobic interior cavity and hydrophilic external surface, are capable of accelerating or inhibiting chemical degradation of organophosphorus pesticides through forming inclusion complexes between CDs and pesticides. This work evaluated the effects of CDs on hydrolysis of malathion in an attempt to assess their potential application in environmental approach. beta-CD and its two derivatives, randomly methylated beta-CD (RAMEB) and hydroxypropyl beta-CD (HP-beta-CD), were tested. It was found that RAMEB could inhibit the hydrolysis of malathion, and this was the function of pH and temperature, the inhibitory effects increase with increasing concentration of RAMEB and elevating temperature between 15 and 35 degrees C. On the other hand, beta-CD and HP-beta-CD have little or no stabilizing effects on malathion at all pH and temperature studied, except that the large concentration of beta-CD and HP-beta-CD can mildly reduce hydrolysis of malathion. Both 2 mol/L and 5 mol/L urea increase the inhibitory effects of RAMEB on hydrolysis of malathion at 25 degrees C, pH 9.0.
    Journal of Environmental Sciences 02/2006; 18(3):572-6. · 1.66 Impact Factor