Prevalence of Common Respiratory Viral Infections and Identification of Adenovirus in Hospitalized Adults in Harbin, China 2014 to 2017

Background: Respiratory infections pose a great challenge in global health, and the prevalence of viral infection in adult patients has been poorly understood in northeast China. Harbin is one of the major cities in northeast China, and more than half of any given year in Harbin is occupied by winter. To reveal the viral etiology and seasonality in adult patients from Harbin, a 4-year consecutive survey was conducted in Harbin, China.Methods: From January 2014 to December 2017, specimens were obtained from adult patients admitted to the Second Affiliated Hospital of Harbin Medical University with lower respiratory tract infections. Sputum samples were examined by direct immunofluorescence assays to detect seven common respiratory viruses, including influenza virus (type A and B), parainfluenza virus (type 1 to 3), respiratory syncytial virus and adenovirus. Adenovirus positive samples were seeded onto A549 cells to isolate viral strains. Phylogenetic analysis was conducted on the highly variable region of adenoviral hexon gene.Results: A total of 1,300 hospitalized adult patients with lower respiratory tract infections were enrolled, in which 189 patients (14.5%) were detected as having at least one viral infection. The co-infection rate in this study was 25.9% (49/189). The dominant viral pathogen from 2014 to 2017 was parainfluenza virus, with a detection rate of 7.2%, followed by influenza virus, respiratory syncytial virus and adenovirus. Based on the climate seasons determined by daily average temperature, the highest overall viral detection rate was detected in spring (22.0%, 52/236), followed by winter (13.4%, 109/813), autumn (11.4%, 13/114) and summer (10.9%, 15/137). Adenovirus type 3 strains with slight variations were isolated from positive cases, which were closely related to the GB strain from the United States, as well as the Harbin04B strain isolated locally.Conclusion: This study demonstrated that common respiratory viruses were partially responsible for hospitalized lower respiratory tract infections in adult patients from Harbin, China, with parainfluenza virus as the dominant viral pathogen. Climate seasons could be rational indicators for the seasonality analysis of airborne viral infections. Future surveillance on viral mutations would be necessary to reveal the evolutionary history of respiratory viruses

Detection of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> in Patient and Mouse Blood and Ticks by a Duplex Real-Time PCR Assay

<div><p>Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, zoonotic infectious diseases caused by <i>Anaplasma phagocytophilum</i> and <i>Ehrlichia chaffeensis</i>, respectively. Early diagnosis is essential for rapid clinical treatment to avoid misdiagnosis and severe patient outcomes. Simple, sensitive and reliable diagnostic methods are urgently needed. In this study, we developed a duplex real-time PCR assay targeting the <i>A. phagocytophilum ankA</i> gene and the <i>E. chaffeensis</i> TRP120 gene, respectively. The lowest limit of detection of the duplex real-time PCR assay was 100 copies of the targeted <i>A. phagocytophilum ankA</i> gene and the <i>E. chaffeensis</i> TRP120 gene per reaction, and the specificity was 100%. Detection in blood DNA samples from the acute stage of illness for 22 HGA cases and 8 HME cases indicated that the duplex real-time PCR assay was more sensitive than the nested PCR assay. The infection of <i>Citellus</i><i>undulatus</i><i> Pallas</i> with <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> was first confirmed in Xinjiang Province and the positive rate was 3.1% for <i>A. phagocytophilum</i>, 6.3% for <i>E. chaffeensis</i> and 3.1% for co-infection with both pathogens. The rates of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> infection of <i>D</i><i>. silvarum</i> ticks collected from Shanxi Province were 8.2% and 14.8%, respectively, and the co-infection rate was 3.3%. The rates of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> infection in <i>H. longicornis</i> ticks collected from Shandong Province were 1.6% and 6.3%, respectively, and the co-infection rate was 1.6%.</p> </div

Pine needle oil induces G2/M arrest of HepG2 cells by activating the ATM pathway

Over the last two decades, inducing DNA damage of cancer cells by natural medicines has become a research hotspot in the field of cancer treatment. Although various natural medicines have anticancer effects, very few studies have been conducted to explore the anti-cancer effect of pine needle oil. In the present study, the role of pine needle oil in inducing G2/M arrest in HepG2 cells was investigated. The data revealed that pine needle oil could induce DNA damage in a dose-dependent manner. In the pine needle oil-treated HepG2 cells, the protein levels of phosphorylated (p)-ataxia-telangiectasia mutated (ATM), γ-H2A histone family, member X, p-p53, p-checkpoint kinase 2 and p-cell division cycle 25C were evidently increased, indicating that pine needle oil facilitated G2/M arrest in HepG2 cells through the ATM pathway. In response to the treatment with pine needle oil, ATM was activated in HepG2 cells, which subsequently phosphorylated downstream targets and induced G2/M arrest. In summary, the data of the present study indicated that pine needle oil induces G2/M arrest in HepG2 cells by facilitating ATM activation

Molecular analysis of Anaplasma phagocytophilum isolated from patients with febrile diseases of unknown etiology in China.

Although anaplasmosis cases have been nationally identified in China, no human isolates of A. phagocytophilum have been obtained, which limits the analysis of any molecular and genetic contributions to patients' severe clinical manifestations and the study of the bacteria's pathogeneses in China. Given this situation, a joint project was conducted in 2009-2010. A total of 421 febrile cases of unknown etiology were collected and the patients' blood samples were collected for laboratory diagnoses including serologic diagnosis based on the four-fold rise in the anti- A. phagocytophilum IgG titer by indirect micro-immunofluorescence assay (IFA), positive PCR assay and confirmation of A. phagocytophilum DNA and positive culture of A. phagocytophilum and confirmed by amplification and sequencing of the 16S rRNA and ank A genes of the A. phagocytophilum isolates. A total of 570 ticks were collected from the patients' domestic animals (456) and from wild fields (114) for culturing and amplifying and sequencing the 16S rRNA gene of A. phagocytophilum. Phylogenetic analyses were performed on the 16S rRNA and ank A gene sequences of the isolates and the ticks tested in the study. A total of 46 (10.9%) confirmed and 16 (3.8%) probable cases were diagnosed and severe clinical features and higher mortality rates were observed in these Chinese patients. Five isolates were obtained and the 16S rRNA genes of the 5 isolates were conserved but variety for ank A genes. Two human isolates and 1 tick isolate from Shandong Peninsula, where all patients exhibited severe clinical manifestations, were grouped as one clan based on the phylogenetic analyses, while 2 other human isolates were clustered in a second clan. 43.5% of H. longicornis were infected with A. phagocytophilum.The present study is the first to obtain clinical isolates of A. phagocytophilum in China. The diversity of the ank A genes of Chinese isolates will help us to further discern the relationship between the variations in the ank A genes and the severity of the disease's clinical manifestations in China

Compare of Laboratory markers between the patients in Shandong Laizhou Bay and the patients in other 3 areas in the study.

<p>MODS, multiple organ dysfunction syndrome; SD, standard deviation; WBC, white blood cell; PLT, platelet; AST, aspartate aminotransferase; ALT, alanine aminotransferase; LDH, lactate dehydrogenase; CK, creatine kinase; BUN, blood urea nitrogen; PTT, activated partial thromboplastin time; HGB, hemoglobin; ESR, sedimentation rate.</p>a<p>p<0.05, HGA patients without MODS vs HGA patients with MODS, according to student's t-test.</p

Map of China and the Bohai Sea Ring areas,Shandong Province.

<p>The Laizhou Bay, Shandong Province (B:areas enclosed by the black circle), where the 2 human isolates (LZ-HGA-agent-3 and LZ-HGA-agent-4) and the tick isolate (LZ- HGA-agent-T1) isolated. The <i>H. longicornis</i> collected in the Laizhou Bay (D and E). Images of HGA patients with eschars from the Laizhou Bay, Shandong Province (F and G).</p

Clinical and laboratory findings and laboratory diagnosis data in 46 confirmed and 16 probable cases in China, 2009–2010.

<p>AST, aspartate aminotransferase; ALT, alanine aminotransferase; LDH, lactate dehydrogenase; CK, creatine kinase; BUN, blood urea nitrogen; CRP, C-reactive protein; HGB, serum albumin; APTT, activated partial thromboplastin time; ESR, erythrocyte sedimentation rate.</p>a<p>: Forty-two serum samples were obtained during the convalescent phase of the illness.</p>b<p>: A total of 25 tick blood lymph sample pools were used to isolate the bacteria.</p

Sequences used in phylogenetic analysis based on the 16S rRNA gene and <i>ank A</i> gene of <i>A. Phagocytophilum</i>.

<p>Sequences used in phylogenetic analysis based on the 16S rRNA gene and <i>ank A</i> gene of <i>A. Phagocytophilum</i>.</p

Phylogenetic tree based on the partial sequences of <i>A. phagocytophilum ank A</i> gene.

<p>Blue sequences:BJ-HGA-agent-1, CZ-HGA-agent-2, LZ-HGA-agent-3, LZ-HGA-agent-4, and LZ- HGA-agent-T1) and some <i>A. phagocytophilum</i> isolates identified in patients, domestic animals, wild animals and ticks from other parts of the world.</p