Identification of a novel coronavirus causing severe pneumonia in human: a descriptive study / 中华医学杂志(英文版)

BACKGROUND@#Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans.@*METHODS@#We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Wuhan Jinyintan Hospital, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed.@*RESULTS@#Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8% to 99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6% to 87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor.@*CONCLUSION@#A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.

Targeted inhibition of Notch1 gene enhances the killing effects of paclitaxel on triple negative breast cancer cells

Objective To study the influence of targeted inhibition of Notch1 gene on the killing effects of paclitaxel on triple negative breast cancer cells. Methods The triple negative [estrogen receptor (ER)/progesterone receptor (PR)/human epidermal growth factor receptor 2 (Her2)] breast cancer cell line MDA-MB-231 and ER/PR/HER-2-positive breast cancer cell line MCF-7 were cultured, transfected with Notch1-siRNA-overexpression plasmid and blank plasmid, and treated with different concentrations of paclitaxel, and then the cell proliferation activity and apoptosis rate as well as the mRNA expression of Caspase-3, Caspase-9 and Bcl-2 were determined. Results Paclitaxel could decrease the MDA-MB-231 and MCF-7 cell proliferation activity as well as Bcl-2 mRNA expression, and increase MDA-MB-231 and MCF-7 cell apoptosis rate as well as Caspase-3 and Caspase-9 mRNA expression in dose-dependent manners; with the same dose of paclitaxel treatment, the inhibitory effects on MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression as well as the promoting effects on MDA-MB-231 cell apoptosis and mRNA expression of Caspase-3 and Caspase-9 were weaker than those on MCF-7 cell; after 0.5 μM paclitaxel combined with Notch1-siRNA treatment, MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression were significantly lower than those after 0.5 μM paclitaxel combined with control plasmid treatment while cell apoptosis rate and mRNA expression of Caspase-3 and Caspase-9 were higher than those after 0.5 μM paclitaxel combined with control plasmid treatment. Conclusions Targeted inhibition of Notch1 gene may enhance the killing effects of paclitaxel on triple negative breast cancer cells by up-regulating the expression of Caspase-3 and Caspase-9 and inhibiting the expression of Bcl-2.

Targeted inhibition of Notch1 gene enhances the killing effects of paclitaxel on triple negative breast cancer cells

OBJECTIVE@#To study the influence of targeted inhibition of Notch1 gene on the killing effects of paclitaxel on triple negative breast cancer cells.@*METHODS@#The triple negative [estrogen receptor (ER)/progesterone receptor (PR)/human epidermal growth factor receptor 2 (Her2)] breast cancer cell line MDA-MB-231 and ER/PR/HER-2-positive breast cancer cell line MCF-7 were cultured, transfected with Notch1-siRNA-overexpression plasmid and blank plasmid, and treated with different concentrations of paclitaxel, and then the cell proliferation activity and apoptosis rate as well as the mRNA expression of Caspase-3, Caspase-9 and Bcl-2 were determined.@*RESULTS@#Paclitaxel could decrease the MDA-MB-231 and MCF-7 cell proliferation activity as well as Bcl-2 mRNA expression, and increase MDA-MB-231 and MCF-7 cell apoptosis rate as well as Caspase-3 and Caspase-9 mRNA expression in dose-dependent manners; with the same dose of paclitaxel treatment, the inhibitory effects on MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression as well as the promoting effects on MDA-MB-231 cell apoptosis and mRNA expression of Caspase-3 and Caspase-9 were weaker than those on MCF-7 cell; after 0.5 μM paclitaxel combined with Notch1-siRNA treatment, MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression were significantly lower than those after 0.5 μM paclitaxel combined with control plasmid treatment while cell apoptosis rate and mRNA expression of Caspase-3 and Caspase-9 were higher than those after 0.5 μM paclitaxel combined with control plasmid treatment.@*CONCLUSIONS@#Targeted inhibition of Notch1 gene may enhance the killing effects of paclitaxel on triple negative breast cancer cells by up-regulating the expression of Caspase-3 and Caspase-9 and inhibiting the expression of Bcl-2.

A research of Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus based on experts' questionnaires / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To establish Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus.</p><p><b>METHODS</b>A Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus was established by literature retrieval and peer review. Two questionnaires were carried out to investigate the confirmation and advice of in-group specialists to key points of the draft including diagnosis, treatment and nursing. Then the procedures were revised accordingly.</p><p><b>RESULTS</b>The recovery rate of complete questionnaires in the 1st survey was 96%. Specialists confirmed more on case history and physical examinations, syndrome differentiation of three syndrome types, treatment of blood deficiency wind dryness syndrome (BDWDS) and nursing. They held different opinions on the outlines, auxiliary examinations, treatment of blood heat induced wind evil syndrome (BHWES) and wind cold and dampness accumulation syndrome (WCDAS), of which the coefficient of variations (CVs) was within 0.1603 -0.2473. The procedures were revised and the 2nd survey was launched. The recovery rate of complete questionnaires in the 2nd survey was 100%. Specialists confirmed more on case history and physical examinations, diagnostic criteria, syndrome differentiation of BDWDS and WCDAS, and treatment of BDWDS, of which CVs was 0. All indicated high agreement and good compliance. The CVs of other items were within 0.0638-0.1439, less than those of the 1st survey. The consistency of experts' opinions were somewhat improved. The contribution by one single item showed less difference in assessing the overall results in the two surveys. A new revision of the procedure was preliminarily established according to results of two surveys.</p><p><b>CONCLUSIONS</b>Experts' activeness, concentration, and coordination were good in the two surveys. They had reached consensus in key points of the draft including diagnosis, treatment, and nursing.</p>

Evaluation of adrenocortical function in children with severe and critical enterovirus 71 infection / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the adrenocortical function in children with severe and critical enterovirus 71 infection by using a high-dose (250 µg) adrenocorticotropic hormone (ACTH) stimulation test. And to at provide experimental basis for glucocorticoid in the treatment of hand-foot-and-mouth disease (HFMD).</p><p><b>METHOD</b>This was a prospective multi-center study which was carried out in PICUs of Beijing Children's Hospital, Zhengzhou Children's Hospital, Kaifeng Children's Hospital and Linyi People's Hospital in Shandong province. Children with severe and critical hand-foot-mouth disease admitted to PICUs of the four hospitals from June 2009 to April 2010 were enrolled in this study, and EV71 virus nucleic acid test and high-dose (250 µg) ACTH stimulation started at the same time. EV71 virus nucleic acid positive 51 cases were eventually enrolled in the study. Cortisol test was performed at baseline (T0) and after high-dose (250 µg) ACTH stimulation at 30 minutes (T30), 60 minutes (T60) in the first 6 hours after admission, but before glucocorticoid was given. The adrenocortical function was evaluated according to ΔTmax [ΔTmax=(T30, T60 maximum)-T0]. Diagnostic criteria of adrenal insufficiency (AI) is increment (ΔTmax)≤9 µg/dl.</p><p><b>RESULT</b>The incidence of AI in 51 cases was 52.94% (27/51). The incidence of AI in severe group was 44.74% (17/38), which was significantly higher in critical group 76.92% (10/13), P<0.05. Of the cases with a pediatric critical illness score (PCIS)≤70, 81.82% (9/11) had adrenal insufficiency, and it was 28.57% (4/14) when PCIS≥90. The incidence of AI was 75% (6/8) and 48.84% (21/43) in death and survivor group respectively, but there were no significant difference between the two groups (P>0.05). Baseline (T0) cortisol in death group was higher than survivor group (P<0.05).</p><p><b>CONCLUSION</b>AI may occur in children with enterovirus 71 infection. The critical enterovirus 71 infection had a high incidence of AI. AI may affect the prognosis of patients with severe and critical enterovirus 71 infection. Exogenous glucocorticoids administration may be considered when AI is identified or highly suspected. The timing, dosage and regimen of glucocorticoid are still unclear. Further animal experiments and clinical trials are needed.</p>

Clinical feature and treatment of 69 Chinese children patients infected with influenza A (H1N1) / 中华儿科杂志

<p><b>OBJECTIVE</b>During the 2009 influenza A (H1N1) epidemic in China, children are the main group among people infected with influenza A (H1N1) virus, but few reports about children are available. The present study aimed to observe the clinical, laboratory features and to analyze therapeutic result.</p><p><b>METHOD</b>The research subject were 93 children infected with influenza A (H1N1), 59 male and 34 female who were treated in Beijing Ditan Hospital from 15 May 2009 to 10 September 2009. The patients' data on symptoms, signs, chest X-ray, blood routine test, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), liver function, renal function, helper T lymphocyte were collected and analyzed. The patients were treated with Oseltamivir, traditional Chinese medicine and symptomatic treatment.</p><p><b>RESULT</b>The main symptoms of children infected with influenza A (H1N1) are fever (84 cases, 90.3%), cough (62 cases, 66.7%), pharyngodynia (36 cases, 38.7%) and expectoration (19 cases, 20.4%) at onset, and fever (59 cases, 63.4%), cough (52 cases, 55.9%), pharyngodynia (23 cases, 24.7%) and expectoration (9 cases, 9.7%) were the mojor symptoms and signs while the patients visited our hospital. The main signs were fervescence, pharyngeal congestion (53 cases, 57.0%), tonsilar swelling (21 cases, 22.6%), and abnormal white blood count (WBC) was found in 32 cases, abnormal ESR in 10 cases, abnormal CRP in 10 cases, abnormal CD4 T lymphocyte count in 19 cases, abnormal liver function and renal function were found in very few patients. After treatment, the febrile duration and time to virus negative in patients treated with oseltamivir alone, traditional Chinese medicine alone, combined oseltamivir and traditional Chinese medicine as well as those who were neither treated with oseltamivir nor traditional Chinese medicine were respectively 1 - 6 days (median 1 day), 3 - 13 days (median 7 days), 1 - 6 days (median 1.5 days), 4 - 11 days (median 8 days), 1 - 5 days (median 1 days), 5 - 14 days (median 8 days), 1 - 5 days (median 2 days), 4 - 13 days (median 8 days).</p><p><b>CONCLUSION</b>Clinical manifestations of 93 children cases were the same as those of adults. The traditional Chinese medicine could improve symptoms of children infected with influenza A (H1N1), but other clinical therapeutic effects need further study.</p>

Clinical characteristics and treatment of severe encephalitis associated with neurogenic pulmonary edema caused by enterovirus 71 in China / 世界急诊医学杂志（英文）

BACKGROUND:Hand-foot-mouth disease has become a major public health issue in children in China. In the present prospective study we investigated the clinical characteristics and emergency management of children with severe encephalitis associated with NPE caused by enterovirus 71. METHODS:The study was conducted in 2 pediatric intensive care units (PICUs) over a 2-month period. Clinical records were reviewed of critically ill children with severe encephalitis associated with NPE caused by EV71 who were admitted to PICUs during the period of May to June 2008 in Fuyang. RESULTS:We reviewed the complete records of 36 children, of whom 23 (63.9％) were male and 13 (36.1％) female. Their age ranged from 4 to 48 months, with an average of 15.8 months. Al children except one were under 3 years of age. The overal mortality in these children was 19.4％. The average duration of critical life threatening signs and symptoms was 2.1 days (12 hours-5 days). Nervous system diseases included brainstem encephalitis in 27 children (75％), brainstem encephalitis associated with myelitis in 6 children (16.7％), and general encephalitis in 3 chidren (8.3％), respectively. In 12 patients of NPE (33.3％) pink or bloody bubble sputum and asymmetric pulmonary edema or hemorrhage was the primary manifestation but no typical exanthema was observed. Five children died of acute onset of NPE and / or pulmonary hemorrhage with rapid progression of cardiopulmonary failure within hours after admission. Therapeutic management consisted of mechanical ventilation and administration of mannitol, methylprednisolone, intravenous immunoglobulin (IVIG) and vasoactive drugs, associated with the need of fluid volume resuscitation in 9 (25％) of the 36 children. CONCLUSIONS:In children less than 3 years of age found to be affected by severe EV71 encephalitis associated with NPE, one fifth may die. The major organ systems infected by severe EV71 include the central nervous system, the respiratory system, and the cardiovascular system. Early diagnosis and evaluation, respiratory support, treatment of intracranial hypertension, and mainttenance of function of the cardiovascular system are the most important therapeutic measures.

Epidemiologic and clinical characteristics of 59 persons suspecting of being infected by the HIV virus despite having repeated negative laboratory findings / 中华流行病学杂志

Objective To describe the epidemiologic characteristics and clinical manifestations of 59 persons recruited via an intemet chat group who complained of AIDS-like symptoms, so as to formulate effective intervention strategies and measures. Methods Case was defined as onset of any three of the following self-reported AIDS-like symptoms in a member of relevant "intemet chat groups": persistent low grade fever, rash, swollen lymph node, fatigue, diarrhea, weight loss and low CD4+T count. We administered an internet-based questionnaire, and invited 59 of the 88 casepersons for voluntary physical examination and laboratory testing. Results The 59 case-persons came from 22 provinces; 54 (91.5 %)were men; the median age was 34 (range: 22-53)years; 84.7% of them had high-risk sexual behaviors before the onset of self-reported symptoms. The median time interval from exposure to onset was 15 d (range: 1-365 d). Blood specimens for all the 59 case-persons were tested negative for HIV and syphilis antibodies. There was also no evidence of Xenotropic murine leukemia virus-related virus infection. One case-person was tested positive for hepatitis C virus antibody. The average CD4'T lymphocyte count was 707/μl. Of the 59 case-persons,57 (96.6%) sought medical care from multiple providers; 40 were diagnosed to have no physical disorders. Conclusion None of the 59 case-persons had any evidence of infection with HIV or any other infectious agents that could explain their self-reported symptoms.

Specific T-cell responses to Mycobacterium tuberculosis protein ESAT-6 in Chinese HIV positive individuals / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To detect TB specific T cell responses by using the recombinant ESAT-6 protein as stimulus in Chinese HIV infected patients.</p><p><b>METHODS</b>ELISPOT-IFN-gamma assay by using the recombinant ESAT-6 protein as stimulus to detect specific T cell responses in HIV+ patients with or without clinical manifestation of TB diseases.</p><p><b>RESULTS</b>Recombinant ESAT-6 protein specific T cell responses show significant high frequencies in both of TB patients with or without HIV infection than that in the healthy control and HIV+ group without clinical TB diseases.</p><p><b>CONCLUSION</b>The ELISPOT-IFN-gamma assay by using recombinant ESAT-6 protein as stimulus could be used in diagnoses of TB infection in Chinese HIV infected patients.</p>

Study on the association of clinical characteristic, CD4+ and level of HIV viral load among 690 initial HIV-infection / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the correlation between CD4+ cell count, HIV viral load (VL) and clinical characteristics among patients when HIV-1 was tested positive and initial AIDS diagnosis was made.</p><p><b>METHODS</b>690 HIV-infected cases from Beijing Di-Tan Hospital were included and under a cross sectional study while SPSS statistical method was used.</p><p><b>RESULTS</b>The 690 HIV-infected cases would include 458 males and 232 females with age range from 2-72 years (mean age as 35.3). The modes of transmission showed that: homosexual contact taking up 17.5% while heterosexual was 16.7%. Most of the homosexual-infected ones lived in Beijing and most of them had bachelor or master's degrees. 19.4% of the transmission happened between heterosexual/bisexual couples, suggesting that HIV was transmitted through the "bridge population" while the rest were infected by contaminated blood/plasma. Many of the cases were identified when they lately visited the pre-operation surveillance point in the hospital. Serious immunodeficiency symptoms or signs were discovered as: CD4+ count < 50 cell/microl, serious opportunistic infections including pneumocystosis pulmonary, cerebral toxoplasmosis and cryptococcal meningitis. Higher frequencies of diseases seen were dermotosis, pneumonia, upper respiratory tract infection, hepatitis and digestive tract moniliasis.</p><p><b>CONCLUSION</b>Because of the late identification of the disease, serious immuo-suppression situation often appeared, suggesting that there was an urgent need to improve STD/AIDS knowledge on those HIV (+) people so they might have an early access to accept medical care.</p>

Association of haplotype formed on HLA-DRB1 and HLA-DQA1 alleles with outcomes of hepatitis B virus infection / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of haplotype of HLA-DRB1 and HLA-DQA1 alleles with outcomes of hepatitis B virus infection in Han population of north China.</p><p><b>METHODS</b>Two hundred and seven chronic hepatitis B (HB) patients, two hundred and twelve chronic asymptomatic hepatitis B virus (HBV) carriers (HBV carrier) and one hundred and forty-eight self-limited HBV infection were investigated for HLA-DRB1 and HLA-DQA1 alleles by sequence specific-polymerase chain reaction (PCR-SSP).</p><p><b>RESULTS</b>The frequency of DRB1*04-DQA1*0301 haplotype was 10.03% in self-limited HBV infection subjects, significantly higher than that in chronic HB patients (3.66%) (P=0.0005)ûthe frequency of DRB1*15/*16-DQA1*0102 haplotype was 6.80% in self-limited HBV infection subjects, significantly higher than 1.94% in chronic HB patients (P=0.0012) and 1.65% in asymptomatic HBV carriers (P=0.0004)ûwhile the frequency of DRB1*04-DQA1*0302 haplotype was 3.10% in chronic HB patients, higher than that in self-limited HBV infection subjects (0.39%) (P=0.0077).</p><p><b>CONCLUSION</b>Individuals with different haplotypes composed of HLA-DRB1 and HLA-DQA1 might have different outcomes of HBV infection.</p>

Association of polymorphisms of human leucocyte antigen -DRB1 and -DQA1 allele with outcomes of hepatitis B virus infection in Han population of north China / 中国医学科学院学报

<p><b>OBJECTIVE</b>To assess the association of polymorphisms of human leucocyte antigen (HLA) -DRB1 and -DQA1 region allele with outcomes of hepatitis B virus (HBV) infection in Han population of north China.</p><p><b>METHODS</b>A total of 207 chronic hepatitis B (HB) patients, 212 chronic asymptomatic HBV carriers (HBV carrier), and 148 self-limited HBV infection were recruited to examine the association between gene polymorphisms and outcomes of HBV infection. Polymerase chain reaction-sequence specific primers (PCR-SSP) technique was used to genotype HLA-DRB1 and HLA-DQA1 loci.</p><p><b>RESULTS</b>The frequency of HLA-DQA1 * 0301 in chronic HB patients (14.81%) was significantly lower than those in HBV carriers (25.24%) and self-limited HBV infection subjects (25.00%) (Pc = 0.002; Pc = 0.007). The frequency of HLA-DQA1 * 0102 in self-limited HBV infection subjects (8.78%) was significantly higher than those in chronic HB patients (2.18%) and HBV carriers (1.89%) (Pc = 0.000; P = 0.000). In addition, the frequency of HLA-DQA1 * 0302 in self-limited HBV infection subjects (4.05%) was significantly lower than that in chronic HB patients (11.41%) (Pc = 0.005). HLA-DQA1 * 0302 was demonstrated to be risk factors of chronic HBV (OR = 3.913, P = 0.0006), while HLA-DQA1* 0102 and HLA-DQA1 * 0301 to be protective factors against chronic HBV (OR = 0.200, P = 0.0004; OR = 0.258, P = 0.0000) after age, sex, smoking and drinking were adjusted by logistic regression analysis. There were positive interactions between drinking and HLA-DQA1 * 0102 [interaction index (II) = 1.49] or HLA-DQA1 * 0302 (II = 12.12). There were negative interactions between drinking and HLA-DQA1 * 0301 (II = 0.78)</p><p><b>CONCLUSIONS</b>The subjects with HLA-DQA1 * 0302 allele have an increased risk to chronic HB infection compared with other subjects without this allele, while HLA-DQA1 * 0301 and HLA-DQA1 * 0102 are associated with HBV clearness. Gene-environment interaction can affect the outcomes of HBV infection.</p>

Difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To clarify the difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome SARS) and common atypical pneumonia.</p><p><b>METHODS</b>Totally 100 patients hospitalized in Beijing Ditan Hospital since March to June 2003 with clinical diagnosis of SARS were involved in this study. These patients courses of disease were over 3 weeks. These patients were divided into two groups, SARS group and common atypical pneumonia group (non-SARS group). The counts of CD3+, CD4+ and CD8+ T-lymphocyte of two groups were systematically recorded and analyzed.</p><p><b>RESULTS</b>Sixty-five of the patients were confirmed to have common type of SARS, including 26 males and 39 females, 50 cases received methylprednisolone treatment. Thirty-five cases had common atypical pneumonia (non-SARS), 21 were males while 14 were females, 20 cases received methylprednisolone treatment. All the cases of two groups were cured in the end. The SARS patients T-lymphocyte counts decreased first and then increased. Before 15 days of disease course, mean CD3+, CD4+, CD8+ T-lymphocyte counts of SARS patients were decreased apparently (694+/-568/microl, 441+/-356/microl, 309+/-462/microl). After 15th day of disease course, the counts gradually returned to normal CD3+, CD4+, CD8+ T-lymphocyte counts of non-SARS patients were normal. Compared with patients of the same group who were not treated with glucocorticoids, T-lymphocyte counts of non-SARS patients treated with glucocorticoids had no obvious difference. But glucocorticoids had some effect on SARS patients recovery of cellular immune function, i.e., it delayed the recovery by about 6 days.</p><p><b>CONCLUSION</b>With or without treatment with glucocorticoids,the lowered CD3+, CD4+, CD8+ T-lymphocyte counts in the early stage are of very important significance in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia.</p>