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To establish a method for simultaneous detection of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3), specific primers and TaqMan probes were designed after sequence alignment according to the specific sequences of PCV2 Cap gene and PCV3 Cap gene on GenBank. By optimizing the reaction conditions, a duplex fluorescence quantitative PCR detection method for simultaneous detection of porcine circovirus type 2 and 3 was established, and the specificity, sensitivity, and reproducibility were tested. Specificity test results showed that in addition to the positive test results for PCV2 and PCV3, tests for PRRSV, CSFV, PPV, PRV, PEDV, and TGEV were all negative with no cross-reaction, indicating its good specificity. Sensitivity test results showed that the minimum detection limit for detection of PCV2 and PCV3 can both reach 10 copies.L-1, indicating its high sensitivity. The coefficient of variation within and between groups of this method was less than 2%, indicating its good stability. A total of 181 pork and whole blood samples collected from Zhejiang Province were tested using the detection method established in this article and the standard common fluorescent PCR detection method. The results showed that the positive rate of PCV2 was 50.83% (92/181), the positive rate of PCV3 was 37.57% (68/181), and the co-infection rate of PCV2 and PCV3 was 12.15% (22/181). The above detection results of ordinary fluorescent PCR were 50.28% (91/181), 36.46% (66/181), and the co-infection rate was 11.60% (21/181). The coincidence rates of the two methods for PCV2 and PCV3 can reach 98.91% and 97.06%, and the coincidence rate for PCV2 and PCV3 mixed infection were 95.45%. In summary, the duplex fluorescence quantitative PCR detection method established in this experiment can distinguish PCV2 and PCV3 rapidly, which can be used for pathogen detection and epidemiological investigation.
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This journal issue includes s of papers presented at the conference. Topics discusses are: stillbirth during a pandemic;analysis of the female genital tract (FGT) metabolome;effectiveness of REGEN-COV antibody combination to reduce risk of hospitalization;patterns of nucleic acid amplification testing;delta variant neutralizing antibody response following maternal COVID19 vaccination;integrated prenatal and hepatitis c virus care increases linkage;extended interval gentamicin dosing in obstetrics;maternal and infant cytomegalovirus detection among women living with HIV.
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Incidences of major feline viral diseases provide basic information for preventing viral disease in cats. Despite the growing interest in feline viral diseases, sero-surveillances have been lacking. In this study, we analyzed the diagnoses of feline viral diseases and conducted a sero surveillance of feline panleukopenia virus (FPV), feline calicivirus (FCV), feline herpesvirus-1 (FHV-1), and feline infectious peritonitis virus (FIPV) in Korean cats. Of the 204 confirmed cases since 2015, the numbers of diagnoses for FPV, FIPV, FCV, feline influenza virus, and FHV-1 were 156, 32, 12, 3, and 1 case, respectively. In total, 200 sera, collected between 2019 and 2021, were screened for the presence of antibodies against FPV, 2 FCVs, FHV-1, and FIPV using a hemagglutination inhibition test and a virus-neutralizing assay (VNA). The overall seropositive rates in cats tested for FPV, the 2 FCVs, FHV-1, and FIPV were 92.5%. 42.0%, 37.0%, 52.0%, and 14.0%, respectively. A low correlation (r = 0.466) was detected between the VNA titers of 2 FCV strains. The highest incidence and seropositive rate of FPV reveal that FPV is circulating in Korean cats. The low r-value between 2 FCVs suggests that a new feline vaccine containing the 2 kinds of FCVs is required.
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Objective: To explore the distribution and correlation of pathogens in the elderly patients with AECOPD, so as to guide the rational use of antibiotics and hormones in clinic. Methods: A total of 111 patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) admitted to Nanjing First Hospital from January 2019 to January 2020 were retrospectively analyzed. The basic data such as eosinophil, neutrophil and lymphocyte count, the levels of C-reactive protein(CRP) and erythrocyte sedimentation rate (ESR)in blood routine examination were collected. Further, the pathogens were qualified by sputum fluorescence quantitative polymerase chain reaction, and the pathogens distribution was analyzed. Results: The level of ESR and the ratio of cardiovascular diseases showed significant differences between the pathogen-positive group and pathogen-negative group. In this study, the top five pathogens in AECOPD patients were EB virus (21.6%), Haemophilus influenzae (19.8%), Streptococcus pneumoniae (17.1%), herpes simplex virus(14.4%), influenza A virus(14.4%). The detection rate of influenza A virus was correlated with influenza B virus and Aspergillus (P < 0.05);The detection rate of respiratory syncytial virus was correlated with Candida, Moraxella catarrholis, Streptococcus pneumoniae and Haemophilus influenzae (P < 0.05);The detection rate of Escherichia coli was correlated with rhinovirus, adenovirus, Klebsiella pneumoniae and Acinetobacter baumannii (P < 0.05);The detection rate of Candida was correlated with that of Moraxella catarrholis and Pseudomonas aeruginosa(P<0.05);The detection rate of human coronavirus was correlated with Haemophilus influenzae, herpes simplex virus and Streptococcus pneumoniae(P < 0.05). Conclusions: AECOPD are mostly induced by different pathogens, especially mixed infection of bacteria and virus. It is helpful to guide the rational use of antibiotics by analyzing the etiological characteristics in the elderly patients with AECOPD.
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The article is devoted to the global problems of modern medicine - HIV infection and the COVID-19 pandemic. The review of the literature highlights current ideas about the pathogenesis and course of COVID-19 in patients with HIV infection, and also touches upon the problems of concomitant pathology and mental health of patients with HIV in the setting of the COVID-19 pandemic. It has been shown that HIV-positive patients are a risk group for the severe course of COVID-19, in particular, individuals with severe immunodeficiency (CD4+ T lymphocytes 200 cells/l) due to the development of synergetic lung damage by SARS-CoV-2 and secondary infectious agents such as cytomegalovirus and Pneumocystis carinii. It has been proven that one of the targets of the SARS-CoV-2 virus is CD4+ T cells, which in COVID-19 leads to a more rapid progression of immunodeficiency in patients with HIV infection and, thus, significantly increases the risk of secondary diseases and death. Particular attention should be paid to middle-aged and elderly people living with HIV, who, compared with HIV-negative patients, are more likely to have concomitant pathology - arterial hypertension, cardiomyopathy and diabetes mellitus, which are the risk factors for severe COVID-19. The results of studies on the effect of antiretroviral drugs on the course of COVID-19 showed that HIV-infected patients receiving tenofovir + emtricitabine have a lower risk of severe COVID-19 and associated hospitalization than patients receiving other HIV treatment regimens. Clinical and preclinical data support the potential use of tenofovir in the treatment of novel coronavirus infection.
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This paper describes the epidemiology, prevalence, transmission, prevention and control of some infectious diseases in companion animals, livestock, wild animals and humans in Ontario, Canada, in 2022, including SARS-CoV-2;Echinococcus multilocularis, Leishmania spp. and SARS-CoV-2;antimicrobial stewardship resources;2 cases of rabid dogs imported from Iran (July 2021 and January 2022);prevalence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriacea, Dirofilaria immitis, Brucella canis, canine parainfluenza and adeno- and herpes viruses in dogs recently imported from Asia;Paragonimus kellicotti lung flukes and Streptococcus equi subsp. zooepidemicus in dogs;African swine fever in pet pigs, backyard pigs and wild pigs and blastomycosis in dogs and humans.
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Herpes zoster is a cluster blister disease that often occurs on the skin of the elderly, immunocompromised individuals and patients with chronic diseases, which is caused by the reactivation of varicella zoster virus dormant in neurons. The persistent pain of herpes zoster tortures patients and is responsible for serious mental and economic burden. With no effective drug treatment for patients, vaccination becomes more important in disease prevention. Now live attenuated vaccine and recombinant protein vaccine with adjuvant are clinically available abroad. Based on the clinical data, recombinant protein vaccine with adjuvant is the preferred one recommended by CDC.In China, many enterprises perform clinical studies on the two vaccines. However, due to the limitation of adjuvant supply, recombinant protein vaccines have not been widely provided. With the global pandemic of COVID-19, mRNA vaccine becomes a hotspot worldwide. Domestic biological enterprises should actively develop new herpes zoster vaccine to fill the vaccine vacancies in China and to enhance market competitiveness. This article briefly reviews the research progress and development of herpes zoster vaccine in various vaccine platforms.
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In the Chinese city of Wuhan at the end of 2019, an infection by an unknown virus began, which with subsequent studies was called SARS-CoV-2, causing a pandemic that has generated the largest crisis worldwide in recent years, causing a large number of deaths, with multiple systemic manifestations but which has also had clinical pictures at the skin level;recently there have been reports of people who had COVID-19 infections and later had skin manifestations due to herpes virus as a co-infection;the most frequent were herpes simplex type 1-2, varicella zoster, herpes zoster and herpes virus 6-7, generating even more complications in patients. Although the pathogenesis of this association is not entirely clear, it is believed to be secondary to the state of immunosuppression induced by SARS-CoV-2, being important that health personnel are informed about this entity that increases mortality.
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Objective: To study the mechanism of Runfei Ningshen Decoction in the treatment of insomnia caused by corona virus disease 2019(COVID-19) by using network pharmacology and molecular docking analysis. Methods: The chemical components and targets of Chinese medicinal materials of Runfei Ningshen Decoction in TCMSP, Batman, and CTD databases were searched. The relevant targets of novel coronavirus pneumonia and insomnia in Disgenet, GeneCards, CTD, and Malacards databases were searched. The component-target-disease network was established by using Cytoscape 3.2.1 software;The protein-protein intereation(PPI) network was constructed in string database. The common targets were enriched by using Cluster Profiler software package in R language software platform. The molecular docking of core targets related to insomnia caused by COVID-19 was carried out by using Discovery Studio 4.0 software. Results: 349 medicinal ingredients in Runfei Ningshen Decoction, 1 904 targets, 1 505 new coronavirus pneumonia-related targets, and 1 337 insomnia-related targets were collected. When the intersection of Venn diagrams were used, 404 common targets were obtained for the 2 diseases. 250 targets were intersected with the 2 diseases, and 33 core targets were screened out by the analysis of the interaction network between targets. Pathway enrichment analysis showed that Runfei Ningshen Decoction mainly acts on AKT1, INS, TP53, IL-6, key targets such as AKT1, INS, TP53, IL-6, JUN, CASP3, TNF, CAT, PTGS2 and CXCL8, which are involved in the important pathway processes such as human cytomegalovirus infection, fluid shear stress, and AGE-RAGE signaling pathways in complications of atherosclerosis and diabetes. The results of molecular docking showed that the core target has a high affinity with beta-sitosterol, 1-methoxy phaseolin, 3'-hydroxy-4'-O-methylglycyrrhizin, and anhydroicariin. The prescription treatment of insomnia caused by COVID-19 may be through the targets such as PTGS2, AR, PPARG, NOS2, HSP90 AA1 and so on. Conclusion: Runfei Ningshen Decoction can treat insomnia caused by COVID-19 by inhibiting IL-6 and TNF-a.
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Objective: To explore the effect and mechanism of Xuanfei Jiere Granules in the treatment of corona virus disease 2019 (COVID-19) complicated with fever. Methods: The effective components of Xuanfei Jiere Granules were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the potential target genes were predicted. The disease targets of COVID-19 complicated with fever were searched by GeneCards and CTD databases, and the common targets were obtained, and then introduced into Cytoscape to construct the "component-target-disease" network. The above 2 common targets were input into the online database of STRING protein interaction, and the results were imported into Cytoscape software to obtain Protein-Protein Interaction(PPI) network;R language was used to analyze the common targets with Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. Results: 84 active components and 82 targets were obtained. The pathway enrichment analysis showed that Xuanfei Jiere Granules mainly acted on protein kinase B(AKT1), tumor protein P53(TP53), interleukin-6(IL6) and other key targets in the treatment of COVID-19 complicated with fever. KEGG pathway enrichment analysis showed that the action was mainly related to the influence of fluid shear stress and atherosclerosis, human cytomegalovirus infection, and AGE-RAGE in diabetic complications and other signal pathways. The results of molecular docking showed that the core target had strong affinity with beta-sitosterol, formononetin, N-trans ferulyl tyramine and so on. Conclusion: This study preliminarily verified that Xuanfei Jiere Granules can play a role in the treatment of COVID-19 complicated with fever through multi-components, multi-targets and multi-pathways.
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This paper describes the clinical signs and use of differential laboratory diagnostic techniques (computed tomography, cytology, histopathology, antigen/antibody detection and polymerase chain reaction) for infectious (viral, bacterial, fungal and parasitic) and non-infectious (inflammatory/immune mediated, neoplastic, cardiac, malformation, foreign body, smoke inhalation, aspiration of caustic material, non-cardiogenic, pulmonary oedema, traumativ, pneumothorax, pulmonary contusions and idiopathic) causes of respiratory diseases in cats and dogs in Ontario, Canada.
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The Indian poultry market is estimated to have an annual growth rate of 8.1% as of today. However, infectious diseases in poultry pose an important constraint in the growth and development of this sector in our region. Among infectious diseases, viral diseases of poultry pose a serious threat to the poultry industry from an economic point of view. Several viral disease outbreaks have been reported by various researchers from different parts of the country. Among the common viral diseases of poultry, incidences of Newcastle disease, Avian Influenza, Fowl Pox, Infectious Bursal Disease, Marek's disease, Infectious Bronchitis, Infectious Laryngotracheitis and Inclusion Body Hepatitis are significant in Assam as well as other parts of India. Thorough epidemiological studies followed by the identification of different serotypes, pathotypes, strains, etc. by genotyping and molecular characterization of viral disease pathogens may lead to ways to control and eradicate the diseases. Importance should be given to maintaining basic preventive measures like biosecurity, farm hygiene, and proper vaccination. In a developing country like India, disease outbreaks can impact the country's economy. In this study, a brief view of the common viral disease of poultry and its diagnosis and control strategies in Assam, India is depicted. However, this review well indicates a plethora of avian diseases that have occurred over the years causing a severe impact on poultry farming as a whole.
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Objective: To elucidate the mechanism of interferon gene stimulating factor (STING) in the anti-pathogenic microbial infection of pigs, so as to further provide a reference for the scientific prevention and control of viral diseases such as porcine transmissible gastroenteritis, epidemic diarrhea and porcine pseudorabies. Method: High-scored targets were found in exons 4 and 8 of STING gene and corresponding sgRNA sequences were designed based on CRISPR/ Cas9 technology. The annealed sgRNAs were linked with the enzyme digested LentiCRISPRV2 carrier with T4 DNA ligase to obtain LentiCRISPRV2-STING-sgRNA lentivirus carrier(STING-sgRNA);Different combinations of STING sgRNA lentivirus carriers, packaging plasmid psPAX2 and envelope plasmid pMD2.G were transfected into 293T cells to obtain lentivirus containing sgRNA and then transduced into 3D4/21 cells. Monoclonal cell lines were obtained by puromycin screening and limited dilution method. The knockout efficiencies of the STING gene were identified by PCR amplification, Sequencing and Western blotting;The effect of STING gene knockout on the expression of type I interferon was verified by real-time fluorescent quantitative PCR. Result: When 293T cells were transfected with different combinations of STING-sgRNA lentivirus carrier and HA-STING over expression vector, the editing effect of STING eukaryotic expression carrier could be detected in cells, and the combination of STING-sgRNA(1+5)lentivirus carrier showed the supreme editing efficiency. Thus, the STING-sgRNA(1+5)lentivirus carrier combined with the packaging plasmid psPAX2 and the envelope plasmid pMD2.G were transfected 293T cells to package lentivirus, and then infected 3D4/21 cells with lentivirus. The results showed that a 3D4/21 cell line with a large deletion of the STING gene(4989 bp)was obtained. The STING protein was not observed by Western blotting, indicating that the STING gene knockout 3D4/21 cells(3D4/ 21-STING-/-)were successfully constructed. The transcription level of IFN-beta in 3D4/21-STING-/- cells decreased significantly (P<0.05) compared with parental cells when stimulated by transfection of Haemophilusparasuis DNA. Conclusion : By applying CRISPR/Cas9 technology, STING gene is successfully knock out in 3D4/21 cells, resulting in loss of function of STING gene;STING knockout leads to the transcription disorder of type I interferon when cells are stimulated by DNA, which also suggests that STING gene may be a key factor in the anti-pathogenic microbial infection of pigs.
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In order to build a specific, sensitive and rapid detection method for PAstV3 detection, the PAstVB gene sequences in Genbank were used and the conserved region in ORFlb was selected to design specific primers and TaqMan probe. Clinical stool samples were collected and preliminary detected by this newly established real-time RT-PCR method after reaction systems and conditions optimization. This detection method established in this study has a good linear relationship with the standard curve, with R2 value up to 0.9971. The sensitivity is 100 times higher than conventional PCR method, The variation co-efficient of in-batch and inter-batch repeatability test is less than 2.0%, indicating good repeatability. The detection results of Clinical samples showed that the positive rate of this method is higher than conventional PCR method. The establishment of this method provides a rapid detection means for PAstV3 laboratory diagnosis and epidemiological investigation.
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Objective: To investigate the clinical. features and treatment of ophthalmopathy secondary to EB virus(EBV) infection in children.
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This proceedings contains s on the proper use of antibiotics, multidrug-resistant infections and new molecules;influenza and COVID-19;osteoarticular infections;and pneumocystis and CMV in the immunocompromised (excluding HIV).
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To establish a J2-KD (knockdown) cell line stably expressing interfered IFITM1 and study the effect of interference with IFITMI gene on the infection of PCV2, PRV and TGEV. Gene cloning tech- niques were used to constructed pLKO. l-EGFP-Puro-IFITMI recombinant vector, which was co-transfected into 293 FT cells with lentiviral packaging plasmids psPAXZ and pMDZ. G to produce green fluorescent protein labeled lentiviruses expression IFITMlshRNA, the viral supernatant was collected at 48 hours after post transfection. J2 cells were infected with the harvested lentiviruses, screened by puromycin and cloned via cell limited dilution. Real-time PCR identify that the cell lines with stable interference with IFITMl gene were obtained, and via MTT method verify that interference with IFITMI expression had no effect on the growth of J2 cells, the successfully constructed J2 stable cell line interfere with IFITMl expression was named as JZ-KD. PRV, PCV2 and TGEV infected J2-KD cells, respectively. Using real-time fluorescence quantitative PCR detect virus replication. The results showed that J2-KD cell line was successfully generated with interfered IFITMl expression;the copy number of PCV2 and TGEV were in- creased, while PRV was decreased in J'Z-KD cell. Indicating that the interference of IFITMI gene expression markedly inhibited the replication of PRV while promoted that. of TGEV and PCV2, providing a basis for further study on the function of porcine IFITMI protein and elucidates its antiviral mechanism.
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Histone deacetylases (HDACs) are a class of key enzymes that regulate epigenetics. There are 5 small-molecule HDACs inhibitors having been approved for anti-cancer therapy on the market. In recent years, there have been more and more studies on the antiviral aspects of HDACs inhibitors. This article classifies viruses into human immunodeficiency virus 1 (HIV-1), new coronavirus (SARS-CoV-2), Epstein-Barr virus (EBV) and other viruses, systematically summarizes the recent advances of antiviral effects of the HDACs inhibitors from the perspective of medicinal chemistry. This review aims to provide the researchers the convenience of accessing the latest advances of the antiviral effects of HDACs inhibitors, and to analyze the challenges and prospects of this field in future drug discovery.
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The infection caused by SARS-CoV-2 may result in a series of skin damages. In addition, some patients report the re-activation of the varicella-zoster virus, which might be related to T cell immune dysfunction caused by SARS-CoV-2 infection. Recently, studies reported herpes zoster occurrence after inoculating the COVID-19 vaccine. At present, the mechanism of interaction between COVID-19, COVID-19 vaccine and herpes zoster remains unclear, and more high-quality studies are required to further define the relationship.