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Objective:To analyze the epidemiological characteristics and genotypes of human rhinovirus (HRV) in patients with upper respiratory tract infection in Qingdao in the winter of 2020.Methods:Throat swab samples were collected from 101 patients with upper respiratory tract infection in Qingdao from November 2020 to January 2021. Quantitative PCR was used to detect 15 common respiratory viruses in the samples. HRV-positive samples were further analyzed with RT-PCR to amplify and sequence HRV VP4/VP2 gene. A phylogenetic tree was constructed based on the sequencing results and homology analysis was conducted.Results:Six common respiratory viruses were detected in the 101 patients. Thirty-four cases (34/101, 33.66%) were single pathogen infection and two cases were multiple infection (2/101, 1.98%). The positive rate of HRV was the highest (21.78%, 22/101). Twenty HRV VP4/VP2 sequences were successfully amplified. Phylogenetic analysis showed that there were 16 strains of HRV-A subtype and four strains of HRV-C subtype and 14 serotypes were involved.Conclusions:HRV was one of the leading viral pathogens causing upper respiratory tract infection in Qingdao in the winter of 2020 and the predominant subtype was HRV-A.
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Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.
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Objective@#To isolate and identify human rhinovirus (HRV) from throat swab samples from patients with acute respiratory infection in Anhui Bengbu.@*Methods@#The throat swab specimens from 108 patients with acute upper respiratory tract infection diagnosed in the Anhui Bengbu were selected as samples. RNA was extracted and detected by HRV universal primers; H1-Hela cells were infected with the positive samples for virus isolation, and HRV VP1 gene amplification was performed and gene phylogenetic tree analysis of the successfully isolated HRV strains was done.@*Results@#Only 5 samples were positive for HRV by the real-time PCR, and only one sample showed significant cytopathic effects after three passages of H1-Hela cells were infected, and the HRV VP1 gene was amplified by RT-PCR in the sample that was named TYZQ201901. The sequence analysis showed that the VP1 nucleotide homology of the strain with other representative HRV A strains was over 95%. The gene phylogenetic tree analysis showed that the strain had the closest genetic distance to the RMH127/2013 strain, and both were on a branch and was confirmed to be HRV type A virus.@*Conclusions@#An HRV type A strain virus was isolated from throat swab samples from patients with acute respiratory infection in Anhui Province by the H1-Hela cells, and the HRV virus separation technology system were initially established.
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Objective@#To isolate, identify and analyze the sex-determining gene fruitless(Anstfru)of malaria-transmitting mosquito Anopheles stephensi.@*Methods@#The full length cDNA of the gene Anstfru was obtained by using bioinformatic and molecular biological method . RT-PCRmethod was used to validate the sex variable splicing pattern and expression time characteristics. The structural features and molecular evolutional features of FRU protein of Anopheles stephensi were analyzed via comparison with FRU protein of known species.@*Results@#The full length of Anstfru gene was isolated and identified, and sex-specific mRNA of the gene could form in female and male mosquitos through variable splicing. The Anstfru began to be expressed from early 1st-2nd stage larvae embryo, the quantity of expression increased subsequently and displayed the highest expression level in adult stage. The FRU protein had the sequence-conservative BTB and zinc finger functional domains.@*Conclusions@#Anstfru gene showed conservative functional domains and sex-determining gene fru expression features in mosquito and further in-depth studies on which will facilitate the application of techniques separating female mosquitos from male mosquitoes, and sterile insect technique (SIT)/technology in prevention and treatment of mosquito-borne infectious diseases.
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Objective To explore the mediation effect of adult attachment on the relationship between childhood trauma and borderline personality disorder(BPD) in recruits.Methods Childhood trauma questionnaire-28 short form (CTQ-SF),adult attachment scale (AAS) and personality diagnostic questionnaire-4+ (PDQ-4+) were administered to 1 750 recruits in the autumn of 2013.Results The scores of physical neglect,emotional neglect,emotional abuse,sexual abuse,physical abuse,attachment anxiety,attachment close and dependence and BPD were 11.05±3.04,10.00±3.54,6.73±2.32,5.92±2.06,5.83± 1.97,3.02 ± ±0.53,3.19±0.44,2.45± 2.04,respectively.BPD had significantly positive correlation with childhood trauma and attachment anxiety (r=0.353,0.284,P＜ 0.01),and significantly negative correlation with attachment close and dependence(r=-0.198,P＜0.01).Childhood trauma,attachment close and dependence and attachment anxiety could positively predict BPD(F=142.172,P＜0.01),which could explain 19.6％ of the total variation.Structure equation model and Bootstrap test showed that childhood trauma not only directly predict BPD in recruits,but also indirectly predict BPD in recruit through attachment close and dependence and attachment anxiety (x2/df=11.472,RMSEA =0.077,NFI =0.943,RFI =0.901,IFI =0.948,TLI =0.902,CFI =0.948),and the mediational path through attachment close and dependence and attachment anxiety with the effect size were 10.8％ and 8.1％,and the total mediational effect size was 18.9％.Conclusion Adult attachment exerts a multiple mediation effect on the relationship between childhood abuse and BPD in recruits.
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The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.
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Humans , China , Ebolavirus , Classification , Genetics , Hemorrhagic Fever, Ebola , Diagnosis , Virology , Laboratories , Workforce , Reference Standards , Laboratory Infection , Quality Control , RNA, Viral , Genetics , Sierra LeoneABSTRACT
Several virus families have been shown to encode microRNAs (miRNAs), which have roles in the infection and replication of viruses in host cells. These virus-encoded miRNAs are identified in double-stranded DNA virus (dsDNA virus) and in several RNA virus families, but not in single-stranded DNA virus (ssDNA virus). We used a bioinformatics approach based on VMir, miRNAFold and MaturePred software to predict virus-encoded miRNA-like small RNAs from the genome of a ssDNA virus: Aedes aegypti densovirus (AaeDV). Northern blotting and stem-loop reverse transcription-polymerase chain reaction (RT-PCR) were used to detect predicted small RNAs. A miRNA-like small RNA termed "AaeDVMD" was identified by stem-loop RT-PCR from predicted candidates. This is the first report demonstrating that a ssDNA virus can encode miRNA-like small RNAs. These data will aid further exploration of the interaction between the AaeDV and its mosquito host.
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Animals , Aedes , Virology , Base Sequence , Computational Biology , Densovirinae , Chemistry , Genetics , Metabolism , MicroRNAs , Chemistry , Genetics , Metabolism , Molecular Sequence Data , RNA, Viral , Chemistry , Genetics , MetabolismABSTRACT
Objective Too investigate the application value of procalcitonin(PCT)and high sensitivity C-reactive protein(hs-CRP)in early diagnosis of neonatal septicemia and disease condition assessment.Methods 48 patients with neonatal septicemia treated in the hospital from November 2011 to May 2013 were collected.The data of PCT,hs-CRP and blood culture were recorded and performed the comparative analysis with the serum PCT,hs-CRP detection results in contemporaneous 48 neonates without septicemia.Results The serum PCT and hs-CRP was 93.75% and 10.42% in the neonates with septicemia,which were signifi-cantly higher than 79.17% and 50% in the neonates without septicemia(P <0.05),the positive rate had statistical difference be-tween the two groups.Conclusion PCT and hs-CRP have remarkable change in the early stage of neonatal sepsis,the combination detection of serum PCT and hs-CRP can be used as the indicators for early diagnosis of neonatal sepsis,moreover the sensitivity and specificity of PCT for diagnosing neonatal septicemia are higher the those of hs-CRP,their combined detection can provide fast and accurate diagnostic basis for clinic.
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The responses of macrophages to Bacillus anthracis infection are important for the survival of the host, since macrophages are required for the germination of B. anthracis spores in lymph nodes, and macrophage death exacerbates anthrax lethal toxin (LeTx)-induced organ collapse. To elucidate the mechanism of macrophage cell death induced by LeTx, we performed a genetic screen to search for genes associated with LeTx-induced macrophage cell death. RAW264.7 cells, a macrophage-like cell line sensitive to LeTx-induced death, were randomly mutated and LeTx-resistant mutant clones were selected. AMP deaminase 3 (AMPD3), an enzyme that converts AMP to IMP, was identified to be mutated in one of the resistant clones. The requirement of AMPD3 in LeTx-induced cell death of RAW 264.7 cells was confirmed by the restoration of LeTx sensitivity with ectopic reconstitution of AMPD3 expression. AMPD3 deficiency does not affect LeTx entering cells and the cleavage of mitogen-activated protein kinase kinase (MKK) by lethal factor inside cells, but does impair an unknown downstream event that is linked to cell death. Our data provides new information regarding LeTx-induced macrophage death and suggests that there is a key regulatory site downstream of or parallel to MKK cleavage that controls the cell death in LeTx-treated macrophages.
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Animals , Mice , AMP Deaminase , Genetics , Anthrax , Pathology , Antigens, Bacterial , Toxicity , Bacterial Toxins , Toxicity , Base Sequence , Blotting, Western , Cell Death , Cell Line , Cell Survival , Cells, Cultured , Exotoxins , Toxicity , Macrophages , Cell Biology , Molecular Sequence Data , Polymerase Chain ReactionABSTRACT
To prepare a large amount of pure alive Schistosoma japonicum eggs, rabbit was infected with 2000 cercariae and its liver was taken aseptically 38-45 days after infection and homogenized. The homogenate was screened through different sieves(60, 120, 200, 300, 360 meshes per inch respectively), and washed with 1.2% NaCl. The eggs and leftover were then digested with 0.25% trypsin for 2 hours, sieved over 360 meshes per inch and washed with RPMI 1640 medium. The collected eggs reached to (95.1?6.4)% of live eggs, with a high efficiency.
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Objective To screen the Schistosoma japonicum female specific expressing genes. Methods{S.japonicum} adult worms were collected from the rabbits’ vein after six-week infection by affusing method. The adult worms were stabilized by RNA-later liquid, the male and female worms were carefully separated with nipper. The high quality total RNA was extracted and mRNA was obtained after purification. Double stranded cDNAs were synthesized after reverse transcription. Female subtractive (female as tester, male as driver) and male subtractive (male as tester, female as driver) cDNA libraries were constructed. The differentially expressed genes were further screened by dot-blot hybridization. The clones were selected and sequenced, which showed apparently higher signals when hybridizing with the female subtracting male probes, than those signals when hybridizing with the male subtracting female probes. The homology of these sequences was searched with BLAST program. The semi-quantitative PCR was applied to test the differential gene expression in female and male adult worms. Result Female subtracting male and male subtracting female cDNA libraries were constructed with SSH technique. After dot-blot hybridization, 50 clones were tested to be the potential female differentially expressed genes and were sequenced. 42 expressing sequence tags (ESTs) were received. After bioinformatics analysis, 17 fragments (about 40