To investigate the efficacy of amiodarone and lidocaine in cardiac arrest patients.

Objective:To analyze the genetic stability of two recombinant rotaviruses (rLLR/NSP3 NLuc) and (rLLR/NSP3 CoV2/RBD) that are inserted and express exogenous genes for continuous passage and proliferation on MA104 cells.Methods:After measuring the titers of two recombinant rotavirus strains, they were transferred to the P2 generation according to MOI0.01. Subsequently, the previous generation of virus lysate was diluted and activated at 1∶100, and MA104 cells were continuously infected for 18 generations (P20). The virus titers of the P1, P5, P10, P15, and P20 generation of cell lysate were measured using indirect immunofluorescence, and RT-PCR identification and dsRNA PAGE silver staining were performed. The luciferase activity of rLLR/NSP3-NLUc was also detected.Results:No inserted fragment loss was found in the recombinant rotavirus rLLR/NSP3 NLuc within 20 generations, with recombinant virus titers ranging from 3.85~5.16 × 10 6 FFU/ml, with strong luciferase signals in each generation. The recombinant rotavirus rLLR/NSP3 CoV2/RBD showed loss of inserted fragments in the 6th generation, with infectivity titers ranging from 2.6 to 3.36 in the first 5 generations of the recombinant virus × 10 6 FFU/ml. Conclusions:The recombinant rotavirus with 582 bp NLuc inserted at the end of the NSP3 gene has good genetic stability, while the recombinant rotavirus with 885 bp RBD inserted at the end of the NSP3 gene was only stable in the first 5 generations, indicating that foreign genes can be inserted at the end of the NSP3 gene of the recombinant rotavirus, and the insert can express, but its stability requires more in-depth research.

Objective:A genome-wide molecular characterization of FJ21351116, a strain of G3P[8]-E2 2021 collected in Fujian, China, was performed.Methods:Whole genome sequencing of FJ21351116 was performed using a high-sensitivity group A rotavirus whole genome sequencing method. Genomic characteriza-tion of the virus was assessed by nucleic acid sequence analysis using MEGA 11.0, Geneious 9.0.2 and DNASTAR software. Neutralization epitopes of VP7 and VP4 (VP8*) were analyzed using BioEdit v. 7.0.9.0 and PyMOL v. 2.5.2.Results:In this study, FJ21351116 was shown to be a G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genotype, and the result of phylogenetic tree showed that the VP7, VP4, VP3, and NSP2-NSP5 genes of the FJ21351116 strain were related to the equine-like DS-1-like G3P[8] genes that have been detected in Japan in recent years. VP6, VP1, VP2, and NSP1 genes are closely related to G2P[4] in most countries, especially in Singapore, suggesting that this strain was formed by genetic reassortment during the evolution of equine-like G3P[8] and G2P[4]. Evolutionary relationships between the VP7/VP4 genes of FJ21351116 and Rotarix and RotaTeq vaccines suggest that the multiple mutations in both VP7 and VP4 (VP8*) neutralizing antigenic epitopes and vaccine amino acid sites. It is hypothesized that the Rotarix and RotaTeq vaccines may be less effective against equine DS-1-like G3P[8] RVA, and the sequence differences with Rotarix are higher than those with RotaTeq.Conclusions:In this study, we found a rare case of DS-1-like G3P [8] RVA strain in China. Currently, horse-like DS-1-like G3P [8] RVA is relatively rare in China and may be poorly protected by vaccine strains, emphasizing the importance of continuous monitoring of RVA strains and the development of efficient and full-coverage RVA vaccines.

Rotavirus is one of the most common pathogens causing severe gastroenteritis in children under 5 years of age in the world. RV vaccination is the most effective measure to control rotavirus diarrhea in infants. The immune effect of RV vaccine is different in countries with different income levels. In this study, the factors affecting the immune effect of RV vaccine are systematically expounded, which provides reference for the research of RV vaccine and the formulation of immune strategy.

Objective:To investigate the effect of microwave on human adenovirus type 2 (HAdV-2) capsid protein in simulated infectious wastes.Methods:Droplets of HAdV-2 virus suspension were added to medical disposable gloves to simulate infectious waste, irradiated under different microwave conditions, the temperature change was recorded, and the irradiated viral supernatant was treated with Dnase I and detected by PCR and qPCR to determine the effect of microwave on the integrity of the viral capsid. ELISA was used to detect the effect of microwave irradiation on the structure of viral hexon protein. The virus was treated alone at the highest temperature during microwave irradiation to investigate whether there were non-thermal effects during microwave disinfection.Results:The maximum temperature during microwave irradiation was 76 ℃, and the PCR and qPCR result showed that Dnase I could significantly damage the viral nucleic acid after microwave irradiation, while the virus control group and heat treatment group were not significantly affected, indicating that microwave irradiation could destroy the integrity of the viral capsid. The result of ELISA showed that microwave irradiation could significantly weaken the binding ability of Hexon protein and antibody, and the result of heat treatment group were similar.Conclusions:Microwave irradiation can destroy the integrity of the HAdV-2 capsid and the structure of Hexon protein, in which the damage to the integrity of the capsid is mainly due to non-thermal effects, and the structural changes of hexon protein are mainly due to thermal effects.

Objective:A recombinant adenoviral vector vaccine based on non-replicating human adenovirus type 5 (Ad5), encoding the conserved domain of respiratory syncytial virus G protein (RSV-G) was constructed. The immunogenicity and protective efficacy of this vaccine were subsequently evaluated in mice.Methods:The recombinant Ad5 vector plasmid (Ad5-Gbcc-Gacc) was constructed by inserted conserved domains of RSV A and RSV B. The recombinant adenovirus Ad5-Gbcc-Gacc was rescued in HEK293A cells. The genome of virus Ad5-Gbcc-Gacc was identified by multi-enzyme digestion, and the expression of Ad5-Gbcc-Gacc was verified by Western blot. Recombinant adenovirus was used to immunize BALB/c mice via intramuscular injection with signal dose, and then challenged with RSV Long strain at week 6. The levels of G specific IgG and antibody subtypes in serum were detected by enzyme-linked immunosorbent assay, the level of neutralizing antibodies was determined by micro-neutralization assay. After challenge, the mice′s weight was recorded daily, the copies of RSV virus in the lung and nasal tissues were detected. Pathological changes in lung tissue were also examined.Results:Western blot and multi-enzyme digestion identification confirmed the successful rescue of the recombinant adenovirus. Ad5-Gbcc-Gacc elicit high titers of specific IgG, robust neutralizing antibodies, and a balanced Th1/Th2 immune response in mice. In comparison to unimmunized controls, mice immunized with Ad5-Gbcc-Gacc reduced the viral copies in both lung and nasal tissue, and exhibited only minimal pathological damage of lung tissue following RSV challenge. In conclusion, Ad5-Gbcc-Gacc induced robust immunogenicity and offers protective effects against RSV infection in murine models.Conclusions:Ad5-Gbcc-Gacc induce robust immunogenicity and can protect mice from RSV challenge, which lays a foundation for further development of RSV vaccine based on G protein.

Objective:To explore the protective effect and safety of RotaTeq vaccine on children with rotavirus gastroenteritis (RVGE) in high mortality areas in the world and guide the correct use of RotaTeq vaccine.Methods:The literature on RotaTeq vaccine in high mortality areas in the world published from February 2006 to December 2021 was searched, screened and sorted out according to the exclusion and inclusion criteria, and the data were analyzed by RevMan 5.3, Stata 14.0 and SPSS 26.0 softwares.Results:A total of 5 reports were enrolled, including 63 974 subjects, including 32 092 subjects in the vaccine group and 31 882 subjects in the placebo group. In high mortality areas, the protection rates of RotaTeq vaccine against RVGE, severe rotavirus gastroenteritis (SRVGE) and very severe rotavirus gastroenteritis (VSRVGE) were VE RVGE=35% (95% CI: 28%-41%), VE SRVGE=51% (95% CI: 33%-65%) and VE VSRVGE=64% (95% CI: 41%-78%). The protection rates of SRVGE in Asia and Africa are VE SRVGE=43% (95% CI: 28%-55%) and VE SRVGE=57% (95% CI: 17%-77%), respectively. There was no significant difference in the incidences of serious adverse events (SAEs) between RotaTeq vaccine group and placebo group ( χ2=2.05, P=0.152). Conclusions:RotaTeq vaccine has a certain protective effect on severe and above RVGE with good safety in high mortality areas in the world.

Objective:To investigate the epidemiological and clinical characteristics of G2P[4] group A rotavirus (RVA) in hospitalized children with diarrhea in China from 2016 to 2019, and to provide data support for the prevention and control of G2P[4] RVA.Methods:The data of viral diarrhea surveillance network in China from January 2016 to December 2019 were collected. A total of 19 667 specimens of hospitalized children with diarrhea under 5 years old were collected from all monitoring provinces, including 5 437 RVA positive specimens. EpiData 3.0 software and Excel 2010 were used for data collection and collation of viral diarrhea monitoring network, and SPSS 26.0 software was used for data analysis.Results:200 G2P[4] RVA specimens were detected from 5 437 RVA positive specimens, and the constituent rate of G2P[4] RVA was 3.68% (200/5 437) There is a statistically significant difference in the constituent ratio of G2P [4] RVA among RVA positive children in different years ( χ2=38.35, P<0.001), months ( χ2=62.69, P<0.001), and ages ( χ2=9.53, P=0.049). There is a statistically significant difference in the constituent ratio of G2P [4] RVA between rural and urban RVA positive children ( χ2=4.01, P=0.045). Compared with non-G2P[4] RVA hospitalized children, G2P[4] RVA hospitalized children had less proportion of respiratory tract infection ( χ2=6.07, P=0.014), G2P[4] RVA hospitalized children had higher proportion of fever ( χ2=6.68, P=0.010), there was no significant differences in diarrhea ( χ2=0.88, P=0.643), vomiting ( χ2=0.23, P=0.629), extraintestinal neurological symptoms ( χ2=0.18, P=0.668), and no significant difference in rash, sepsis and other complications ( χ2=0.45, P=0.504). Conclusions:The epidemic trend of G2P[4] RVA in China gradually decreased from 2016 to 2019, and the autumn and winter were G2P[4] RVA seasonal peaks. And the peak age was 24-36 months. There were a higher infection risk in rural areas, and fever was more than other genotypes.

Objective:To investigate the sequence characteristics and evolutionary pattern of a strain of G1P[8] genotype group A rotvirus (RVA) SC18511073 in China and to determine the differences between SC18511073 and the antigenic epitopes of RotaTeq? and Rotarix? vaccines.Methods:RT-PCR amplification of 11 segments of SC18511073 was performed using reverse transcription-polymerase chain reaction (RT-PCR), and the typing was conducted by online RVA automatic typing tool. DNAstar5.1 and Mega11.0 software were used to analyze the homology and genetic evolution of the 11 segments.Results:The genotype constellation of SC18511073 is G1P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1, and NSP4 is the E2 genotype. The VP7 and NSP3 segments of SC18511073 had high homology with the 2018 Sichuan epidemic G1P[8]-E1 strain, and the phylogenetic tree showed that they were located in the same branch. The remaining nine gene segments all had high homology with the G9P[8]-E2 type prevalent in China and were attributed to the same evolutionary branch. SC18511073 differs from RotaTeq? and Rotarix? by a total of 5 amino acid sites on 7-1a and 7-2 of VP7, and differences in the 8-1 and 8-3 regions of VP8 * antigen epitopes. Conclusions:The 2018 RVA strain SC18511073 in China is a rare G1P[8]-E2 type, which is a new strain generated by re-assortment of VP7 and NSP3 segments during the co-infection process of G1P[8]-E1 and G9P[8]-E2 RVA strains. SC18511073 has amino acid site changes in antigenic epitopes on VP7 and VP4 with RotaTeq? and Rotarix?.

Objective:To study the inactivation effects of microwave on human adenovirus 2 (HAdV-2) in simulated infectied wastes, and to explore its molecular mechanism.Methods:25 μl of HAdV-2 virus suspension was dripped with medical disposable gloves, masks and cotton swabs to simulate infectied wastes, and irradiated under different microwave conditions: gloves and masks were irradiated for 30 s, 60 s, and 90 s at 300 W, 500 W, and 700 W, respectively. Cotton swabs irradiate 60 s, 90 s, 120 s at 500 W and 700 W respectively. Temperature changes were recorded, and the inactivation logarithmic values were calculated by the 50% endpoint method to evaluate the microwave inactivation effects, and the proliferation ability of the virus was detected by qPCR. The damage of Penton, Fiber, Hexon and E2 B genes was detected by PCR. The virus was treated with the highest temperature of 76 ℃ during microwave irradiation to study whether there was non-thermal effect during microwave disinfection. Results:After microwave irradiation of infectied waste, the temperature of masks and gloves carriers rises rapidly, with the highest temperature of 76 ℃. The temperature of the cotton swab carriers rose slowly, and the highest temperature is 65 ℃. The inactivation effect of microwave on HAdV-2 was positively correlated with microwave power and irradiation time. In the mask and glove group, microwave power of 700 W irradiated for 60 seconds, and the inactivation logarithm value could reach 3.0, In the cotton swab group, microwave power of 700 W irradiated for 120 seconds, and the inactivation logarithm value was still less than 3.0. This indicated that there were differences in the conditions for microwave inactivation of the virus in different carriers. The qPCR result showed that microwave irradiation could weak the proliferation ability of the virus. Microwave irradiation had no effect on the virus's Penton and Fiber genes, but caused some damage to the Hexon and E2 B genes. The inactivation effect of individual heat treatment on HAdV-2 was weaker than that of microwave irradiation, and there was no damage to the Hexon, Penton, Fiber, and E2 B genes. This indicated the presence of non thermal effects during the microwave inactivation process. Conclusions:Microwave irradiation can inactivate HAdV-2 in simulated infectied wastes through thermal and non-thermal effects, and its damage to viral DNA is one of the mechanisms of virus inactivation.