Based on the isolation and identification of Nosema ceranae(N.ceranae)in honeybees,this study optimized the synthesis method of nano-FeS derived from albumen and explored in vitro and in vivo antifungal effects against N.ceranae.Pathogens were isolated from infected honeybee colonies and identified as N.ceranae using morphological and molecular biology techniques.In vitro experiments were conducted to confirm the antifungal effects of nano-FeS against N.ceranae and elucidate the mechanism.In vivo experiments were carried out to validate the therapeutic effects of nano-FeS against N.ceranae infection.Nano-FeS was synthesized using the solvothermal method with an optimal scheme determined through orthogonal experiments,with an average par-ticle size of 75 nm.Flow cytometry and fluorescence staining experiments confirmed that nano-FeS induced apoptosis and necrosis in N.ceranae.After N.ceranae was exposed to nano-FeS,intracellu-lar iron accumulation,disruption of the glutathione and glutathione peroxidase antioxidant system,and subsequent ROS accumulation were observed,ultimately leading to lipid peroxidation of cell membranes.In vivo experiments demonstrated reduced mortality and decreased spore counts in the midgut of honeybees fed with nano-FeS.Transcriptome analysis and qPCR revealed the impact of nano-FeS on gene expression in the N.ceranae infected honeybee midgut.This study presented a promising alternative antifungal agent for N.ceranae infection in honeybees and elucidated the an-tifungal mechanism of nano-FeS related to ferroptosis.Additionally,the study found a positive cor-relation between the mass concentration of nano-FeS and its antifungal effectiveness against N.ceranae.

Objective To establish a method for concentration determination of menthol in the nasal in -situ gel spray. Methods Gas chromatography (GC) was performed on the column of Agilent DB-WAX capillary column (30m ×0.25 mm ×0.25μm), with the column temperature of 120 ℃.With nitrogen gas used as carrier gas , hydrogen flame ion detector (FID) was used, the detection temperature was 260℃, injection temperature was 220℃, injection volume was 1μl and column flow rate was 1.0 ml/min. Results For menthol, there was good linear relationship within the range of mass concentration at 0.02~0.20 mg/mL (r=0.9991). Precision, reproducibility and stability test were all lower than 2%, the sample recovery rate was 95.1%~103.4%and relative stand-ard deviation (RSD) was 2.3%(n=6).Conclusion The established method had the following advantages:easy operation, high re-liability and good reproducibility , and it could be used for the content determination of menthol in the nasal in -situ gel spray .

Objective To obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying methord.Methord The capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with the usage of the favorite codons in E.coil so as to enhance its protein expression in prokaryotic expressing system.The protein was purified by Ni-NTA column,and its antigenicity was determined by Western Blot.Then establish ELISA to detect the specific anti-VP2 IgG antibodies against HBoV1 and 2 in healthy children aged 3 - 6 years in Nanjing,China.Results The recombinant protein 6 × His-VP2 was produced in a larger quantity at 25℃ induced by IPTG (lmmol/L) over night and purified by Ni-NTA column.Seropositive rates of HBoV1and 2 were 62.2％ and 55.5％ and their mixed seropositivity was 37％.Conclusion The optimizing expression of the capsid protein VP2 from human Bocavirus constructed successfully and get a high yield under certain conditions.The established ELISA could be used to further analyze seroepidemiology of HBoV in china.