Broad SARS-CoV-2 cell tropism and immunopathology in lung tissues from fatal COVID-19

Background Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection in patients with Coronavirus Disease 2019 (COVID-19) prominently manifests with pulmonary symptoms histologically reflected by diffuse alveolar damage (DAD), excess inflammation, pneumocyte hyperplasia and proliferation, and formation of platelet aggregates or thromboemboli. However, the mechanisms mediating these processes remain unclear. Methods We performed multicolor staining for viral proteins, and lineage cell markers to identify SARS-CoV-2 tropism and to define the lung pathobiology in postmortem tissues from five patients with fatal SARS-CoV-2 infections. Findings The lung parenchyma showed severe DAD with thromboemboli in all cases. SARS-CoV-2 infection was found in an extensive range of cells including alveolar epithelial type II/pneumocyte type II (AT2) cells (HT2-280), ciliated cells (tyr--tubulin), goblet cells (MUC5AC), club-like cells (MUC5B) and endothelial cells (CD31 and CD34). Greater than 90% of infiltrating immune cells were positive for viral proteins including macrophages and monocytes (CD68 and CD163), neutrophils (ELA-2), natural killer (NK) cells (CD56), B-cells (CD19 and CD20), and T-cells (CD3{varepsilon}). Most but not all infected cells were positive for the viral entry receptor angiotensin-converting enzyme-2 (ACE2). The numbers of infected and ACE2-positive cells correlated with the extent of tissue damage. The infected tissues exhibited low numbers of B-cells and abundant CD3{varepsilon}+ T-cells consisting of mainly T helper cells (CD4), few cytotoxic T cells (CTL, CD8), and no T regulatory cell (FOXP3). Antigen presenting molecule HLA-DR of B and T cells was abundant in all cases. Robust interleukin-6 (IL-6) expression was present in most uninfected and infected cells, with higher expression levels observed in cases with more tissue damage. Interpretation In lung tissues from severely affected COVID-19 patients, there is evidence for broad SARS-CoV-2 cell tropisms, activation of immune cells, and clearance of immunosuppressive cells, which could contribute to severe tissue damage, thromboemboli, excess inflammation and compromised adaptive immune responses.

Mutations, Recombination and Insertion in the Evolution of 2019-nCoV

BackgroundThe 2019 novel coronavirus (2019-nCoV or SARS-CoV-2) has spread more rapidly than any other betacoronavirus including SARS-CoV and MERS-CoV. However, the mechanisms responsible for infection and molecular evolution of this virus remained unclear. MethodsWe collected and analyzed 120 genomic sequences of 2019-nCoV including 11 novel genomes from patients in China. Through comprehensive analysis of the available genome sequences of 2019-nCoV strains, we have tracked multiple inheritable SNPs and determined the evolution of 2019-nCoV relative to other coronaviruses. ResultsSystematic analysis of 120 genomic sequences of 2019-nCoV revealed co-circulation of two genetic subgroups with distinct SNPs markers, which can be used to trace the 2019-nCoV spreading pathways to different regions and countries. Although 2019-nCoV, human and bat SARS-CoV share high homologous in overall genome structures, they evolved into two distinct groups with different receptor entry specificities through potential recombination in the receptor binding regions. In addition, 2019-nCoV has a unique four amino acid insertion between S1 and S2 domains of the spike protein, which created a potential furin or TMPRSS2 cleavage site. ConclusionsOur studies provided comprehensive insights into the evolution and spread of the 2019-nCoV. Our results provided evidence suggesting that 2019-nCoV may increase its infectivity through the receptor binding domain recombination and a cleavage site insertion. One Sentence SummaryNovel 2019-nCoV sequences revealed the evolution and specificity of betacoronavirus with possible mechanisms of enhanced infectivity.

Construction of a self-regulated learning structure model for clinical medical undergraduates in the stage of professional courses / 中华医学教育探索杂志

The purpose of this study is to investigate the basic situation of self-regulated learning among clinical medical undergraduates in the stage of professional courses and construct a structural model suitable for them to carry out self-regulated learning . Zimmerman's self-regulated learning concept framework was determined as the theoretical basis of this study. A questionnaire survey was performed to identify five main dimensions affecting self-regulated learning including expression of learning intention, self-efficacy, adjustment of learning behavior, management of curriculum strategies, and adaption of learning environment, and then, Amos 24.0 software was used to construct the structural model and draw the conclusion that self-regulated learning in this group centers on the regulation of learning behavior, with learning intention and self-efficacy as sub-centers.

Study on relationship between lower extremity venous thrombotic disease and seasons / 中国中西医结合急救杂志

Objective To investigate the relationship between the onset of lower extremity venous thrombotic disease and seasons.Methods A retrospective study was conducted, 300 patients with lower extremity venous thrombotic disease admitted to HandanCity Hospital of Traditional Chinese Medicine (TCM) from August 2012 to February 2014 were enrolled, the incidences and TCM syndromes of patients with lower extremity venous thrombotic disease in different seasons were observed, and the pathogenesis and relationships between the types of TCM syndrome and seasons were analyzed.Results There were 142 patients with lower extremity superficial thrombophlebitis, and 158 cases with lower extremity deep venous thrombosis, the incidence of lower extremity venous thrombotic disease in spring was significantly higher than that in summer and autumn [32.8% (86/262) vs. 21.3% (54/254), 18.4% (50/272), bothP 0.01); while the incidence of lower extremity venous thrombotic disease in winter was significant higher than those in summer and autumn (allP < 0.01). The incidences of damp and heat downward flow type in autumn and summer were increased compared with those in winter and spring [55.6% (30/54), 60.0% (30/50) vs. 20.0% (22/110), 23.3% (20/86), allP < 0.01], while the incidences of lower extremity venous thrombotic disease with damp heat and stasis syndrome in winter and spring were increased compared with those in summer and autumn seasons [80.0% (88/110), 76.7% (66/86) vs. 44.4% (24/54), 40.0% (20/50)].Conclusions The incidence of lower extremity venous thrombotic disease is related to seasons, and the onset is high in winter and spring, damp heat and stasis syndrome being the main type; according to different seasons, clinical treatment can direct to different pathogenic factors to adopt different preventive measures interfering with the patient's constitution in order to eliminate or reduce the risk factors, achieving the effect of the disease prevention.

The identiifcation analysis of rhinoceros horns products based on DNA barcoding 12s rRNA / 中国法医学杂志

Objective Through the DNA barcoding 12s rRNA sequences alignment and analysis of several rhinoceros horns products involved in cases to analysis the application feasibility of 12s rRNA in the rhinoceros horns products’ species identification. Methods Use rhinoceros horns products in 3 cases as materials, total DNA were extracted with improved method, PCR ampliifcation the DNA barcoding. Results The alignment and analysis of sequences show that 12s rRNA could identify rhinoceros horns products at the species level. Conclusion The DNA barcoding 12s rRNA could be used as a new way to identify the rhinoceros horns products which can’t be identiifed with morphological characteristics, provide a reliable basis for the qualitative and sentencing of cases.

Establishment of a national teaching center for virtual simulation experiment on disaster medicine / 中华医院管理杂志

Presented in the paper are the necessity,general ideas and principles of building a teaching center for virtual simulation experiment on disaster medicine,covering the teaching modules, capability objectives and education resources deployment among other basics of such a center.The authors propose to build a comprehensive platform for teaching by experimentation,integrating basic clinical skills training to trainings targeted to disaster rescue in view of actual needs in experiments and teaching.This way resources can be shared between the experiment center website and virtual simulation teaching software,promoting a regular,standardized and scientific development of disaster medicine in China.

Application of PDCA circle model in medical college students' clinical practice training and evaluation / 中华医学教育探索杂志

In order to strengthen the management of clinical teaching, improve the teaching quality of clinical practice, the advantages of the traditional medical education, teaching reform and medical simulation were further integrated, using PDCA (Plan-Do-Check-Action) cycle model in medi-cal college students' clinical practice training and evaluation. The skills of training and evaluation were optimized and practiced by thechain model of teaching-assistance-guidance to improve the train-ing quality of interns, and make continuous improvement from the angle of teaching, learning and management and with PDCA circulation mode. Practice shows that the application of PDCA circulation management has greatly improved teachers' teaching quality and their own quality, and the students' satisfaction of practice teaching management and clinical teachers has also significantly increased. The effect is obvious.

Methods and practice of systematic clinical first-aid skills training for community doctor / 局解手术学杂志

Objective To observe the effect of clinical first-aid skills training for community doctor. Methods Effect of systematic clinical first-aid skills training and traditional teaching methods were scored by the questionnaire survey in order to compare their difference. Results Through systematic clinical first-aid skills training, the score on contacting the actual work, improving the diagnosis and treatment, analyzing and problem-solving skills, enhancing learning initiative, participation, scientific learning method and self-learning ability, impro-ving clinical thinking, strengthening team spirit and humane education were significantly improved compared to that of the traditional teaching methods (P<0. 01). Conclusion The systematic clinical first-aid skills training is an effective way for community physicians.

Retrospective on the construction and practice of a state-level emergency medical rescue team.

For the past few years, disasters like earthquakes, landslides, mudslides, tsunamis, and traffic accidents have occurred with an ever-growing frequency, coverage, and intensity greatly beyond the expectation of the public. In order to respond effectively to disasters and to reduce casualties and property damage, countries around the world have invested more efforts in the theoretical study of emergency medicine and the construction of emergency medical rescue forces. Consequently, emergency medical rescue teams of all scales and types have come into being and have played significant roles in disaster response work. As the only state-level emergency medical rescue force from the Chinese People's Armed Police Forces, the force described here has developed, through continuous learning and practice, a characteristic mode in terms of grouping methods, equipment system construction, and training.

Murine gammaherpesvirus-68 ORF38 encodes a tegument protein and is packaged into virions during secondary envelopment

Tegument is the unique structure of a herpesvirion which occupies the space between nucleocapsid and envelope. Accumulating data have indicated that interactions among tegument proteins play a key role in virion morphogenesis. Morphogenesis of gammaherpesviruses including Kaposi's sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV) is poorly understood due to the lack of efficient de novo lytic replication in cell culture. Murine gammaherpesvirus-68 (MHV-68) is genetically related to these two human herpesviruses and serves as an effective model to study the lytic replication of gammaherpesviruses. We previously showed that ORF33 of MHV-68 encodes a tegument protein and plays an essential role in virion maturation in the cytoplasm. However, the molecular mechanism of how ORF33 participates in virion morphogenesis has not been elucidated. In this study we demonstrated that ORF38 of MHV-68 is also a tegument protein and is localized to cytoplasmic compartments during both transient transfection and viral infection. Immuno-gold labeling assay showed that ORF38 is only present on virions that have entered the cytoplasmic vesicles, indicating that ORF38 is packaged into virions during secondary envelopment. We further showed that ORF38 co-localizes with ORF33 during viral infection; therefore, the interaction between ORF38 and ORF33 is conserved among herpesviruses. Notably, we found that although ORF33 by itself is distributed in both the nucleus and the cytoplasm, in the presence of ORF38, ORF33 is co-localized to trans-Golgi network (TGN), a site where secondary envelopment takes place.

Role of tegument proteins in herpesvirus assembly and egress

Morphogenesis and maturation of viral particles is an essential step of viral replication. An infectious herpesviral particle has a multilayered architecture, and contains a large DNA genome, a capsid shell, a tegument and an envelope spiked with glycoproteins. Unique to herpesviruses, tegument is a structure that occupies the space between the nucleocapsid and the envelope and contains many virus encoded proteins called tegument proteins. Historically the tegument has been described as an amorphous structure, but increasing evidence supports the notion that there is an ordered addition of tegument during virion assembly, which is consistent with the important roles of tegument proteins in the assembly and egress of herpesviral particles. In this review we first give an overview of the herpesvirus assembly and egress process. We then discuss the roles of selected tegument proteins in each step of the process, i.e., primary envelopment, de-envelopment, secondary envelopment and transport of viral particles. We also suggest key issues that should be addressed in the near future.

Clinical Analysis on 425 Cases of POEMS Syndrome / 中国康复理论与实践

@#ObjectiveTo summarize the clinical features of reported 425 domestic cases of POEMS syndrome, and its diagnosis and treatment.MethodsClinical characteristics of 425 POEMS cases were summarized with meta analysis.ResultsPOEMS syndrome usually occurred in middle-age males, and its onset age ranged from 11 to 74 years, averaged 43 years. Clinical display was characterized by a combination of polyneuropathy, organomegaly, endocrinopathy, M-protein and skin changes. It usually occurred with osteomyeloma. Most patients died of serious infections, respiratory insufficiency or heart failure. The treatment with glycocorticosteroid had led to short-time improvement.ConclusionThe patients in our country more present as peripheralneuropathy, skin change and edema, whereas hepatomegaly, splenomegaly, erectile dysfunction, breast enlargement of males, amenorrheal, pleural effusion and ascites are next.

Detection and significance of apoptosis related factors in the primary human embryonic pancreatic islet cells / 第三军医大学学报

Objective To explore the relationship between TGF-?, P21ras, c-fos and the apoptosis of human embryonic pancreatic islet cells. Methods Semi-quantitative RT-PCR and immunohistochemistry (SABC method) were used to detect TGF-?, P21ras, c-fos expression at different growth periods of the primary human embryonic pancreatic islet cells and human fibroblast cells. Results P21ras expression was weak in every period of the pancreatic islet cells and significantly weaker than the fibroblast cells at exponential growth period and was stronger than in incubation period and retention period (P

Effects of U50,488H on L-type calcium current in the normal and hypoxic rat ventricular myocytes / 解放军医学杂志

Objective To explore the nechanism of the interaction between K-opioid receptor and ? adrenal receptor. Method The effects of u50 488H on L-type calcium currents in the normal and hypoxic rat ventricular myocytes were studied by using whole-cell patch clamp technique. Results The basal as well as Isoproterenol-stimulated I_ Ca,L were inhibited by U50,488H in a dose-dependent manner in normal rat ventricular myocytes. In the hypoxic rat ventricular myocytes,the inhibitory effect of U50,488H was decreased. U50,488H had no significant effect on Forkolin-stimulated I_ Ca,L . Conclusion The results indicated that the negative modulation of ?-opioid receptor on ?-adrenoceptor was attenuated in the hypoxic ventricular myocytes,and the target of U50,488H on ?-adrenergic system might be situated between ?-adrenoceptor and adenylate cyclase.

Effects of genistein on the proliferation of cardiac fibroblasts / 中国药理学与毒理学杂志

To study the effect of genistein on the proliferation of cardiac fibroblasts(CF), CFs were cultured from neonatal rat hearts, DNA synthesis of the cells was determined by incorporation of ［3H］TdR into DNA, the cell cycle was measured by flow cytometric analysis. Genistein(0.5－50 μmol*L-1) attenuated 2.5% fetal calf serum-induced proliferation of CF in concentration-dependent manner. Genistein(50 μmol*L-1) arrested CF cell progression at G2/M phase. The results suggest that genistein be a potential substance for treatment of cardiac fibrosis.

Protective role of NGF on cerebral ischemic reperfusion injury and its effect on Bax expression / 临床神经病学杂志

Objective To study the protective role of nerve growth factor (NGF) on cerebral ischemic reperfusion injury and its effect on Bax expression. Methods The models of focal cerebral ischemic reperfusion in rats were made by occlusion middle cerebral artery using an intraluminal filament method. NGF was given through vena caudalis. The expression of Bax was studied by immunohistochemistry method and the change of brain was observed using pathological method.Results Bax positive cells in the cortex and striatum areas in ischemia-reperfusion group (115.25?15.07) were more than those in NGF treatment group (70.00?12.77) ((P

Anti-arrhythmic effects of U50,488H involving the inhibition of Na~+ current in ischemia/reperfusion rat heart / 中国药理学通报

Aim To investigate the effects of U50,488H(a selective ?-opioid receptor agonist)on ventricular arrhythmias induced by myocardial ischemia and reperfusion in rats and to elucidate their mechanisms.Methods The contents of CK(creatine phosphokinase)and LDH(lactate dehydrogenase)were measured; The isolated heart was perfused using langendorff equipment. Heart rate(HR), arterial blood pressure(ABP), left ventricular pressure (LVP), cardiac function (?dp/dtmax), rate of ventricular tachycardia(VT)and ventricular fibrillation(VF)were examined in rats in vitro. The incidence of ventricular arrhythmias and arrhythmia score were also determined; The change of sodium currents (INa) induced by U50,488H was detected by whole-cell recording mode using patch clamp.Results ① In comparison with I/R group, the contents of CK、LDH in plasma of rats in U50,488H+I/R group were significantly lowered(P

Mediative effect of ?-opioid receptor on blood pressure in the rats / 中国药理学通报

Aim To investigate the effects of U50488H,a selective ?-opioid receptor agonist,on the blood pressure in the rats and explore their mechanisms.Methods Heart rate(HR),arterial blood pressure(ABP),left ventricular pressure(LVP),contractive function(+ dp/dt_(max)) and diastolic function(-dp/dt_(max)) were examined in rats.Physiological experimental technique was used to collect urine and to determine the volume of urine output;isolated artery perfusion technique was used to investigate the direct action of U50488H on abdominal aorta of rats.Results HR,ABP,LVP and ?dp/dt_(max) in rats were decreased with the administration of U50488H;the urine volume increased significantly with the administration of U50488H.U50488H induced a dose-dependent vasodilation in the aortic artery.These effects of U50488H were totally abolished by nor-BNI,a selective ?-opioid receptor antagonist.Conclusion Stimulation of ?-opioid receptor with U50488H depresses the blood pressure mainly by reducing the strength of cardiac muscle,enhancing the urine volume and relaxing the vessel.

Antiarrhythmic effect and underlying mechanism of ?-opioid receptor in rats with myocardial ischemia and reperfusion / 中国药理学通报

Aim To investigate the anti-arrhythmic effect and mechanism of ?-opioid receptor during myocardial ischemia and reperfusion in rats,and to initially determine the regulation of U50488H(U50,a selective ?-opioid receptor agonist) to angiotensinⅡ(AngⅡ),endothelin(ET) and nitric oxide(NO) in rats.Methods Rats were randomly divided into 7 groups,i.e.,control group,ischemia/reperfusion group(I/R),U50488H+I/R group,PTX group(PTX,a Gi/o proteininhibitor),Glib group(glibenclamide,a K_(ATP) channel blocker),Che group(chelerythrine,a selective PKC inhibitor),and Gen group(Genistein,a Tyrosine kinase inhibitor) respectively.The arrhythmia occurrence and score in different groups were observed and counted.The contents of AngⅡ,ET and NO in plasma of rats were also examined.Results ① Compared with I/R group,the arrhythmia score of U50+I/R group was significantly decreased.The effect of pared with I/R group,the arrhythmia score of U50+I/R group was significantly decreased.The effect of glibenclamide and chelerythrine respectively,the anti-arrhythmic effects induced by U50488H in the rats during myocardial ischemia and reperfusion were significantly attenuated or even completely blocked.③ The anti-arrhythmic effects of U50488H were not significantly affected by pretreatment with genistein.④ In comparison with normal rats,the contents of AngⅡand ET in plasma of I/R group were significantly increased,but the content of NO was decreased.With the administration of U50488H,the contents of AngⅡand ET in plasma of rats in U50488H+I/R group were significantly decreased.Meantime the content of NO was increased.Conclusions ① U50488H-induced anti-arrhythmic effects in the rats with myocardial ischemia and reperfusion are mediated by ?-opioid receptor.The signaling pathway may be related with(Gi/o,) PKC,and K_(ATP) channel.② The activation of ?-opioid receptor may elicit anti-arrhythmic effect through the down-regulations of AngⅡ or ET and up-regulation of NO in plasma of rats.