Quantification of complete viral particles in inactivated avian influenza virus antigen by high performance size exclusion chromatography coupled with multi-angle laser light scattering / 生物工程学报

We developed a method for accurate quantification of the intact virus particles in inactivated avian influenza virus feedstocks. To address the problem of impurities interference in the detection of inactivated avian influenza virus feedstocks by direct high performance size exclusion chromatography (HPSEC), we firstly investigated polyethylene glycol (PEG) precipitation and ion exchange chromatography (IEC) for H5N8 antigen purification. Under the optimized conditions, the removal rate of impurity was 86.87% in IEC using DEAE FF, and the viral hemagglutination recovery was 100%. HPSEC was used to analyze the pretreated samples. The peak of 8.5-10.0 min, which was the characteristic adsorption of intact virus, was analyzed by SDS-PAGE and dynamic light scattering. It was almost free of impurities and the particle size was uniform with an average particle size of 127.7 nm. After adding antibody to the IEC pretreated samples for HPSEC detection, the characteristic peak disappeared, indicating that IEC pretreatment effectively removed the impurities. By coupling HPSEC with multi-angle laser scattering technique (MALLS), the amount of intact virus particles in the sample could be accurately quantified with a good linear relationship between the number of virus particles and the chromatographic peak area (R2=0.997). The established IEC pretreatment-HPSEC-MALLS assay was applied to accurate detection of the number of intact virus particles in viral feedstocks of different subtypes (H7N9), different batches and different concentrations, all with good applicability and reproducibility, Relative standard deviation < 5%, n=3.

Clinical observation on hormone replacement in treatment of perimenopausal syndrome / 中国医师进修杂志

ObjectiveTo study the changes of related hormone levels using hormone replacement therapy in patients with perimenopausal syndrome,and its clinical significance.Methods Eighty-five patients with perimenopausal syndrome received continuous sequential estrogen and progesterone treatment.Daily oral conjugated estrogen 0.625 mg,28 days as one cycle.In each cycle the 19-28th d combined with medroxyprogesterone acetate 4 mg,once a day,a total of 3 cycles.Observed the changes of hormone levels before and after treatment.ResultsThe levels of estradiol and progesterone were obviously increased after treatment compared with those before treatment [ (83.6 ± 1.9) ng/L vs.(10.0 ± 1.1 ) rng/L,(0.32 ± 0.10)μ g/L vs.(0.22 ± 0.20) μ g/L,P ＜ 0.05 ],and follicles stimulating hormone,luteinizing hormone,total cholesterol and triglyceride were obviously decreased [ (40.8 ± 11.0) U/L vs. (57.1 ± 10.8) U/L, (24.9 ±13.6) U/L vs.(46.2 ± 13.8) U/L,(4.6 ±0.9) mmol/L vs.(6.1 ± 1.0) mmol/L,(1.1 ±0.2) mmol/L vs.( 1.6 ± 0.1 ) mmol/L,P ＜ 0.05 ].Body mass index was no obviously different[ (23.9 ± 1.9) kg/m2 vs.(22.5 ± 2.2)kg/m2,P ＞ 0.05 ].ConclusionApplication of hormone replacement in treatment of perimenopausal syndrome is safe.