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ObjectiveTo investigate the changes in the pathogen spectrum of viral diarrhea in local pediatric inpatients as well as any variations in genotypes of major pathogens during the COVID-19 control period. MethodsFecal samples were collected from the children <5 years who were hospitalized due to acute gastroenteritis in a pediatric hospital in Shanghai. PCR test was carried out to detect rotavirus， norovirus， sapovirus， astrovirus and enteric adenovirus， and then genotyping was performed for major pathogens. ResultsOut of 546 samples， 37.55% tested positive for virus with the following positive rate ranking： norovirus GⅡ （22.16%）， group A rotavirus （16.12%）， astrovirus （2.93%）， enteric adenovirus （2.38%）， sapovirus （0.92%） and norovirus GⅠ （0.18%）. The predominant genotype within norovirus GⅡ were GⅡ.4［P31］ and GⅡ.4［P16］ with a proportion of 24.79% and 14.05% respectively. The detection rate of GⅡ.4［P31］ dropped significantly over the 2-year period （χ2＝16.140，P<0.001）. In addition， an emerging rotavirus genotype G8P ［8］， which was rarely found nationally， was discovered for the first time locally with an increasing proportion， accounting for 7.95% of all rotavirus positive cases. Phylogenic analysis demonstrated that the representative strains of this genotype were genetically closer to the DS-1-like G8P ［8］ strain found in Southeast Asia. ConclusionThe changes in the prevalence of various norovirus genotypes together with the emergence of rare rotavirus genotype in the local area illustrate the importance of continuous monitoring of viral diarrhea and genotyping of key pathogens. Increased local activity of the rare genotype also adds new parameters in the efficacy evaluation of marketed vaccines and development of potential new vaccines in near future.
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AIM:To investigate the effects of Chinese traditional medicine-selected recipe Q0409 on the ability of learning and memory in SAM-P/8 mice. METHODS:Total 91 mice (4-month-old SAM-P/8 mice, SAM-R/1 mice and Kunming mice) were used in the study, in which the male and female animals were labeled separately. According to the performance of Morris water maze test, the mice were divided into 5 groups randomly. The mice were fed with different drugs or distilled water for 60 d (from 4 months to 6 months). The mice were fed with the drugs from 61 d to 65 d, and 1 h later each time, the Morris water maze test was carried out. After this Morris test were finished at 65 d, the mice were killed immediately and their hippocampal tissues were isolated. Half of the hippocampal tissues were added with precooled normal saline and made into 10% (g/mL) homogenate for detecting the protein content and acetyl cholinesterase (AChE) activity. The other half was fixed with 4% paraformaldehyde and embedded with paraffin for immunohistochemical staining of amyloid β-protein (Aβ). RESULTS:Compared with model group, the results of navigation training and spatial probe training in Morris water maze test were significantly improved (P<0.05), and the activity of AChE in the hippocampal ho-mogenate was significantly decreased (P<0.05) in Q0409 treatment group. No difference in Q0409 group was observed compared with control group and positive drug (huperzine A) group. Immunohistochemical staining showed no typical "se-nile plaques" in the male mice of Q0409 group, while there was shallower and smaller brown staining in the hippocampus of the female mice of Q0409 group. The positive area of Aβ deposition decreased in the CA1 area of hippocampal tissues in Q0409 group. These results were similar to those in positive drug group. CONCLUSION:Q0409 improves the ability of learning and memory in SAM-P/8 mice, which is related to the inhibition of AChE activity and the reduction of Aβ protein deposition in the hippocampus. The effects is similar to those of huperzine A.
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La importancia de determinar el sexo en especies monomórficas que viven en cautiverio conlleva a estrategias de apareamiento a fin de poder incentivar la reproducción. Existen métodos no moleculares de reconocimiento pero son factibles en edad adulta y mucho más difícil cuando son polluelos. Mediante la técnica molecular de PCR, se amplificó el gen de la Cromo Helicasa de Unión al ADN empleando los cebadores, 2550F/2718R. Por este método molecular se consiguió determinar el sexo en 19 de 23 especies de las cuales 3 son ejemplares de: Amazona aestiva, 1 de Pipile grayi, 1 de Bubo virginianus, 4 de Ara chloropterus, 6 de Crax fasciolata, 2 de Caracara plancus, 3 de Chauna torquata, 1 de Cariama cristata y 2 de Cairina moschata del Centro de Investigación de Animales Silvestres de ITAIPU binacional, del lado paraguayo. Nuestros resultados sugieren que el par de cebadores 2550F/2718R podría ser de utilidad para determinar el sexo en las especies estudiadas en el país.
In monomorphic species living in captivity, determining the sex is important because it may allow choosing mating strategies to enhance reproduction. Sex can be determined either by non-molecular recognition methods, which are only feasible in adult birds, or by molecular techniques, which may allow sexing monomorphic species at young ages. Thus, in this study, we used molecular techniques such PCR to amplifying the CHD gene by using 2550F/2718R primers. This method allowed us to determine the sex in 19 out of 23 bird specimens from the Centro de Investigación de Animales Silvestres (CIASI) of the ITAIPÚ BINACIONAL at Paraguayan side. The specimens belonged to the following species: Amazona aestiva (3), Pipile grayi (1), Bubo virginianus (1), Ara chloropterus (4), Crax fasciolata (6), Caracara plancus (2), Chauna torquata (3), Cariama cristata (1) and Cairina moschata (2). The 2550F/2718R primers were useful for determine the sex of the studied species.
الموضوعات
Animals , Birds , Molecular Diagnostic Techniques , Polymerase Chain Reaction , Sex Determination Analysisالملخص
As ORFs I e IV do genoma do HTLV-1 codificam, respectivamente, as proteínas p12/p8 (acessória) e Tax (regulatória). p12/p8, de 99 aminoácidos, pode ser clivada em sua extremidade amino terminal gerando a proteína p8. A primeira clivagem proteolítica de p12 remove o sinal de retenção ao RE, enquanto a segunda clivagem, gera o produto de 8kDa, referido como p8. p12 localiza-se no sistema de endomembranes, residindo em RE e aparato de Golgi, enquanto p8 dirige-se para a membrana plasmática, onde é recrutada para a sinapse imunológica, através da ligação com o receptor de células T (TCR), além de participar da sinapse virológica e da formação de conduítes. A proteína Tax, por outro lado, atua como transativador transcricional do HTLV-1, sendo referida também na indução da expressão de diversos genes celulares, aumentando a proliferação e a migração das células infectadas. Na via de transporte de vesículas secretórias, vesículas produzidas como pós-Golgi são transportadas ao longo do citoesqueleto por motores celulares. A Miosina-Va, um motor não convencional, transporta diversos cargos, incluindo vesículas secretórias, vesículas sinápticas e de retículo endoplasmático. Outra proteína relacionada ao citoesqueleto é a Paxilina, que atua como molécula adaptadora nas adesões focais e cuja expressão está aumentada em indivíduos TSP-HAM...
HTLV-1 ORFs I and IV encode respectively p12/p8 (accessory protein) and Tax (regulatory protein). The 99 amino acid p12 protein can be proteolytically cleaved at the amino terminus to generate the p8 protein. The first proteolytic cleavage removes the ER retention/retrieval signal at the amino terminus of p12, while the second cleavage generates the p8 protein. The p12 protein localizes to cellular endomembranes, within the ER and Golgi apparatus, while p8 traffics to lipid rafts at the cell surface and is recruited to the immunological synapse upon T-cell receptor (TCR) ligation, virological synapse and conduits. Tax on the other hand acts as viral transactivator and induces expression of many cellular genes, increasing proliferation and migration of infected cells. In secretory vesicle transport, vesicles produced as post-Golgi are moved along the cytoskeleton by motor proteins. The unconventional myosin motor, Myosin-Va, moves several cargoes including secretory vesicles, synaptic vesicles, and the endoplasmic reticulum. Another cytoskeleton associated protein is Paxillin, an adapter on focal adhesions which expression is increased in TSP-HAM patients...
الموضوعات
Humans , Paxillin/biosynthesis , Paxillin/toxicity , Paxillin/ultrastructure , Gene Products, tax/analysis , Gene Products, tax/immunology , Gene Products, tax/isolation & purification , Gene Products, tax/blood , Gene Products, tax/chemical synthesis , Human T-lymphotropic virus 1/immunology , Human T-lymphotropic virus 1/pathogenicityالملخص
Rotaviruses are important enteric pathogens for humans and animals. Group A rotaviruses (RV-A) are the most common agents of severe gastroenteritis in infants and young children and vaccination is the most effective method to reduce RV-A-associated diseases. G1P[8], the most prevalent RV-A genotype worldwide, is included in the RV-A vaccine Rotarix®. The discrimination between wild-type G1P[8] and vaccine G1P[8] strains is an important topic in the study of RV-A epidemiology to manage outbreaks and to define control measures for vaccinated children. In this study, we developed a novel method to segregate the wild-type and vaccine strains using restriction endonucleases. The dsRNA from the Rotarix® vaccine was sequenced and the NSP3 gene was selected as the target gene. The vaccine strain has a restriction pattern that is different than that of wild-type RV-A G1P[8] isolates after digestion with the restriction endonuclease BspHI. This pattern could be used as a marker for the differentiation of wild-type G1P[8] strains from the vaccine strain.
الموضوعات
Humans , Feces , Rotavirus Vaccines , Rotavirus , DNA Restriction Enzymes , Genotype , Polymorphism, Restriction Fragment Length , Rotavirus Infections , Rotavirus , Vaccines, Attenuatedالملخص
Objective To investigate the effect of fast-aging on the excitability of hippocampus CA1 neurons in mouse and the possible interaction between fast-aging and hippocampus. Methods Using brain slice and extracellular recording technique to record the firing of hippocampus CA1 neurons in fast-aging(SA M-P/8) and normal control mice, after preprocessing, neural firing train were obtained. Using neural firing rate and inter-spike-inter to investigate interaction between fast-aging and hippocampal neural firing. Results The neural firing rate of hippocampus CA1 neurons in fast-aging mice is (1.052±0.364) Hz(n=14), while the neural firing ratein normal control mice is (4.416±1.306) Hz(n=22). In fast-aging mice, 80.5% inter spike intervel(ISI) is longerthan 1sec, but in normal control mice, 95.6% ISI is shorter than 0.5sec. Conclusion The decreased firing rate of hippocampus CA1 and longer ISI observed in the fast-aging mice indicates that fast-aging significantly inhibit hippocampal CA1 neurons excitability.
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Background: Rotavirus strain (KH0118) is used as the primary material to produce original rotavirus vaccine strains with the symbol of G1P8 MS. According to the World Health Organization\u2019s standard, the strain is needed to evaluate the stability of gene throughout analysis of gene and amino acid sequence during vaccine production. Objective: To determine the sequence of genes 4 (VP4), 6 (VP6), 9 (VP7) and 10 (NSP4) with base pair correlative 855:866:1345:745 of seed lot system and vaccine of G1P8 strain and to evaluate the stability of seed lot system during vaccine production. Subject and methods: ARN was divided from the original strain of rotavirus vaccine G1P8 MS, rotavirus vaccine productive strain (G1P8 WS) and rotavirus vaccine (G1P8 VX). Then using primer pairs to determine gene sequence VP4, VP6, VP7, NSP4 and comparing gene and amino acid sequence of the seed lot system. Results and Conclusion: The study demonstrated that, there was no difference for the nucleotide and amino acid sequence from the original strain during production of rotavirus vaccine G1P8 KH0118.
الموضوعات
Rotavirus Vaccinesالملخص
Background: In recent years, the rotavirus diarrhea increased significantly in Vietnam, an estimated 50%-70% of children\u2019s hospitalization for diarrhea in 2005. The virus subtype causing disease mainly in Vietnam today was not only G1, G2, G4, like other countries in the world but also the rare strain was G9. Objectives: to isolate G9 rotavirus strains on MA104 cell culture; to selecting G9 rotavirus which developing well on MA104 cells. Subjectives and Method: an experimental research in the laboratory. 20 stool samples derived from 20 children with acute diarrhea caused by rotavirus G9P8 (using RT-PCR method). The samples were processed according to standards of the Centers for disease control and prevention (CDC), Atlanta, USA. Results: eight out of 20 human rotavirus strains G9P8 were positive on Ma104 cell suspension after 3 consecutive cultures (OD indexes of these sample were over 0.100,sample No 2 had the highest OD (1.347)). Sample No 2 was chosen for the first time cloning (25 clones, 17/25 with OD>0.100). And clone No 16 was selected for the second purifying (25 clones, 24/25 with OD>0.100). Ten out of 25 clones in the second time were adapted on monolayer Ma104 cell culture and only clone No 14 and No 18 with highest OD (2.648 and 2.644, respectively) will be used for next studies. Conclusions: cloning method was a basic method for adapting rotavirus clone on cell cultured. Rate of G9P8 rotavirus strain which isolated on Ma104 cells\ufffd?suspension was relatively high. Among the second time clones, 2/10 samples adapted on Ma104 cells with high OD.
الموضوعات
Cell Culture Techniques , Rotavirusالملخص
Background: Rotavirus vaccines used to prevent acute diarrhea among children under 5 years old. In Vietnam, with the assistance of the World Health Organization (WHO), Centers for disease control and prevention (CDC), Atlanta, United States, Ministry of Health and Ministry of science and technology researched to create rotavirus master seed G1P8 which would use to produce Rota virus in Vietnam. Objectives: to evaluate titre and safety of rotavirus master seed G1P8 in vitro and in experimental animals. Subjectives and Method: experimental studies in vitro and experimental animals. Rotavirus master seed G1P8 (KH0118) series 1 (MS-P5) and series 2 (MS-P5) produced in 2005, preserved 80oC, were determined by immunofluorescence method (with unit of FFU/ml). The strains were tested in experimental animal\u2019s safety like the safe requirements of World Health Organization on live-attenuated vaccine (OPV). Results: no detection for exotic virus strains in virus suspension by PCR and titres of rotavirus master seed were similar and were more than 107 FFU / ml. 10 intervened rabbits were healthy life during follow up period, 8/10 these rabbits increased weight after follow up period, only two rabbits reduced weight but increase of mean weight of all 10 rabbits was 106%. 5 intervened guinea-pig increased mean weight (158%) and healthy during follow-up period. 4 experimental white mice increased mean weight (195%), higher than control groups. It was safety in experimenting on baby mice. Conclusions: two lots of rotavirus master seed G1P8 produced in POLYVAC had high titre and safety both in vitro and in experimental animals.
الموضوعات
Rotavirusالملخص
Rotavirus is the main cause of acute viral gastroenteritis in children under 5 years old. The study is to sequence nucleotides and amino acids of VP4, VP7, NSP1, and NSP4 genes of 5 passages of human rotavirus strain G1P8. The number of nucleotide mutations of VP4, VP7, NSP4 genes occuring among the passages were 3, 1, and 3, respectively. All these mutations resulted in changes in amino acid composition. No mutation was found in NSP1 gene.
الموضوعات
Humans , Child , Rotavirus , Genesالملخص
AIM: To investigate the differences of Alzheimer's disease(AD)-related parameters in the SAM-P/8, the SAM-R/1 and the Kunming mice.METHODS: The changes of ethology, neurobiochemistry (true choline esterase, TchE), ultrastructure and gene expression(gene chips) were determined in mice of three groups: SAM-P/8 mice (n=14), SAM-R/1 mice (n=14) and Kunming mice (n=14), which were 6 months old[weight(20?5)g].RESULTS: The SAM-P/8 mice had the inabilities of learning and memory compared with the SAM-R/1 mice and the Kunming mice (P