We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.

Objective To investigate the effects of ephedrine on the expression levels of brain-derived neurotropic factor (BDNF) and postsynaptic density protein 95 (PSD95) and synapsin1 in PC12 cells, and to explore the mechanism of ephedrine cytotoxicity on PC12. Methods After PC12 cells were treated with different concentration of ephedrine, the cell survival rate was measured by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The morphology changes of PC12 cells were observed by an inverted microscope. Western blot was used to detect the protein expression levels of BDNF, PSD95 and synapsin1 in PC12 cells. Results Ephedrine decreased the viability of PC12 cell in a concentration-dependent manner,with an IC₂₅ and IC₅₀ of 0.536 mmol and 2.8 mmol, respectively, for PC12 cell death. As ephedrine concentration increased, PC12 cells became smaller in size, with blurred boundary blurred, reduced synapses and shorter axon lengths. The expression levels of BDNF and PSD95 increased significantly. Meanwhile the expression level of synapsin1 decreased. Conclusion The mechanism of ephedrine cytotoxicity on PC12 may be related to the expression levels of BDNF, PSD95 and synapsin1.

Human parainfluenza virus (HPIV) is important respiratory pathogen that could cause bronchitis and pneumonia in infants, young children and other immunocompromised populations. At present, there are no vaccines or drugs approved for specific prevention and treatment of HPIV infection. In this review we introduce the development of HPIV vaccines from five categories including inactivated vaccine, live attenuated vaccine, subunit vaccine, mRNA vaccine and combined vaccine.

OBJECTIVE To compare the efficacy of the novel Huoxue plaster and the original powder in the treatment of chronic soft tissue injury, and to provide experimental basis for the development and clinical application of hospital preparations. METHODS According to the different drug administration, mice were assigned to the blank group, the matrix group, the positive control group(voltaren group), the original powder raw product group, the original powder product group and the novel plaster group respectively. The writhing inhibition rate, licking time, analgesic rate and pain threshold were measured by the writhing test, formaldehyde pain test, electrical stimulation test and hot plate test. The purpose of these was to assess disparities in the drugs for analgesic purposes. The mice ear swelling test and capillary permeability test were used to determine the swelling degree, swelling inhibition rate and exudation inhibition rate to compare the differences in anti-inflammatory effects. A rat model of chronic soft tissue injury was established. The changes of calf circumference, soft tissue injury appearance score, hemorheology and histopathological observation and inflammatory factor content were compared before and after the reform of prescription and dosage form. RESULTS The novel Huoxue plaster had a significant inhibitory effect on the inflammatory pain induced by acetic acid and formaldehyde in mice, and there was no significant difference compared with the original powder. The analgesic effect of 60 min after administration was better than that of 90 min and 120 min after administration, and there was no significant difference compared with the original powder. The novel Huoxue plaster and the original powder had no significant effect on hot plate pain threshold. The novel Huoxue plaster had a significant anti-inflammatory effect, which could reduce the degree of ear swelling induced by xylene and reduce the inflammatory exudation of capillaries induced by acetic acid, and there was no significant difference compared with the original powder. The novel Huoxue plaster could significantly reduce the circumference and injury symptom index of the swollen leg in rats with chronic soft tissue injury, improve blood circulation and histomorphology, and significantly reduce the contents of PGE2, TNF-α and IL-1β in the injured soft tissue, and there was no significant difference compared with the original powder. CONCLUSION The novel Huoxue plaster has significant analgesic and anti-inflammatory effects, and can be used to treat chronic soft tissue injury. The efficacy of the novel Huoxue plaster is basically the same as that of the original powder.

OBJECTIVE To investigate the transfer and diffusion of berberine hydrochloride(BBH), the main active component of Coptis and Phellodenticum in the system of carboxymethyl chitosan(CMCS)-sodium alginate(SA). METHODS CMCS and SA were stirred in a certain proportion, and D-gluconolactone(GDL) was added to form polyelectrolyte hydrogel. Rheometer was used to study the rheological properties of CMCS-SA hydrogel, including the elastic modulus G′ and the viscous modulus G′′. A BBH diffusion model for CMCS-SA hydrogel was designed, and the relevant rules of BBH permeation through CMCS-SA polyelectrolyte hydrogel were observed by UV-VIS. RESULTS The elastic modulus G′ of the hydrogel was measured when the ratio of CMCS to SA was 3∶1, 2∶1, 1∶1, 1∶2, and 1∶3. When the ratio of CMCS to SA was 1∶1, G′ was the highest, and the crosslinking strength of the hydrogel was the highest. The cumulative amount of BBH transfer was measured by the BBH transfer model, and the diffusion of BBH in CMCS-SA hydrogel was fitted as the skeleton dissolution by Peppas equation, indicating that BBH dissociation and the transfer efficiency increased as the amino group of CMCS decreased or the carboxyl group of SA increased. The elastic modulus G′ of CMCS-SA hydrogel increased with the increase of GDL content. The reason was that the binding force between CMCS and SA molecules gradually increased with the decrease of pH, and the crosslinking degree of the hydrogel was enhanced. When ratio of fixed CMCS to SA was 1∶1 and the GDL content was 0.15 g·mL−1, the formability of CMCS-SA hydrogel was good. In addition, when BBH was transferred in hydrogel with different concentrations of GDL, the transfer efficiency increased with the increase of GDL content. When BBH was delivered in different thickness hydrogel, the delivery efficiency of CMCS-SA hydrogel increased with the decrease of thickness. CONCLUSION The CMCS-SA hydrogel system, as a potential drug carrier for traditional Chinese medicine extracts such as BBH, is expected to serve as a gel carrier for transdermal drug delivery.

Gel materials have advantages such as comfortable touch, good air permeability, high viscoelasticity, etc., so they are widely used in modern transdermal drug delivery preparations such as patches, gel agents and film agents. With the innovative development of medical materials, many new gel materials with excellent functions have emerged, which has expanded the application scope of transdermal drug delivery preparations and optimized their properties. The classification, properties and application of gel materials in transdermal drug delivery system were reviewed, and the latest research progress was summarized, so as to provide reference for the innovative application of gel materials in transdermal drug delivery system.m.

Objective To analyze the antigenicity,genetic characteristics and variation of the hemagglutinin(HA) protein of influenza A(H1N1)pdm09 virus circulating in Shanghai during 2018-2019 influenza surveillance year.Methods Hemagglutinin inhibition test was performed to analyze the antigenicity of eighty-four influenza A(H1N1)pdm09 virus strains isolated in Shanghai from April 2018 to March 2019.Sixty-five influenza virus strains isolated from different districts of Shanghai were sequenced and analyzed.Results Clade 6B.1 H1N1 virus was the predominant strains circulating in Shanghai.A few epidemic strains belong to the 6B.2 branch.The similarities of clade 6B.1 nucleotide sequences compared with the vaccine strain A/Michigan/45/2015 were 97.1％-98.8％.The homology with the newly recommended vaccine strain A/Brisbane/02/2018 were 97.5％-99.4％.Mainly fifteen amino acids had mutated in the HA protein sequence,and three mutations,S91R,S181T and T202I were involved in three different epitopes which indicated that the antigenic drift had occurred in the influenza virus.Conclusions The majority of influenza A(H1 N 1)pdm09 subtype virus strains circulating in Shanghai were well matched with the vaccine strain A/Michigan/45/2015 recommended by WHO.It is necessary to continue strengthening the surveillance on influenza virus variation to improve the efficacy of influenza vaccines.

ObjectiveTo determine the genomic characteristics of a subgenus B human adenovirus strain isolated in Shanghai in 2021. MethodsAn adenovirus type 55 strain was isolated and identified from a patient with acute hemorrhagic conjunctivitis （AHC）. Complete genome of the strain was obtained using the next-generation sequencing （NGS）. Phylogenetic trees were reconstructed based on the sequences of Hexon， Fiber， Penton and complete genome to genomically characterize this strain. ResultsPhylogenetic analysis based on the complete genome classified this strain （MH2021001） into subgenus B， subspecies B2 of HAdV-55. Hexon gene of MH2021001 had close phylogenetic relationship with HAdV-11， while Fiber and Penton genes had close relationship with HAdV-14. The MH2021001 showed high nucleotide identity with currently prevalent HAdV⁃55 strains （>99.90%）. The complete genome had 99.96% nucleotide identity to the 73-GD_CHN_2016 strain isolated in Guangdong. In addition， the amino acid sequence of MH2021001 had several substitutions in regions coding for E1B， L4， E3 and L5. ConclusionThis strain has been classified to HAdV-B55. No recombination event is identified in the complete genome. Due to multiple amino acid substitutions， the biological characteristics of the strain need to be further identified.

ObjectiveTo isolate and study the biological characteristics of severe acute respiratory syndrome coronavirus 2 （SARS-CoV-2） from feces of coronavirus disease 2019 （COVID-19） patients. MethodsVero E6 cells were used for virus isolation and the isolated strains were tested by nucleic acid test， immunofluorescence test， virulence test and whole genome sequencing. 50% tissue culture infective dose （TCID₅₀） was calculated after the cell cultures of each generation were collected ResultsEight fecal specimens were inoculated with Vero E6 cells after treatment and cultured for 48 h. One specimen showed obvious cytopathic effect on Vero E6 cells. One SARS-CoV-2 out of 8 fecal samples from COVID-19 patients were isolated， and separation rate was 12.5%. The TCID₅₀ of P1， P2 and P3 were 10^4.0/0.2 mL， 10^4.5/0.2 mL and 10^4.75/0.2 mL， respectively. Only one of the 8 stool samples had SARS-CoV-2 virus replication and amplification， and the Ct value of the nucleic acid detection was about 10. The sequence of the isolation was more than 99.99% homologous with that of Wuhan-Hu-1（GenBank MN908947）. ConclusionThe SARS-CoV-2 strain is isolated from the fecal samples of COVID-19 cases and is confirmed by genomic sequencing and immunofluorescence test， which indicates the presence of live virus in feces of COVID-19 cases.