Network-based identification and pharmacological targeting of host cell master regulators induced by SARS-CoV-2 infection

Precise characterization and targeting of host cell transcriptional machinery hijacked by SARS-CoV-2 remains challenging. To identify therapeutically targetable mechanisms that are critical for SARS-CoV-2 infection, here we elucidated the Master Regulator (MR) proteins representing mechanistic determinants of the gene expression signature induced by SARS-CoV-2. The analysis revealed coordinated inactivation of MR-proteins linked to regulatory programs potentiating efficiency of viral replication (detrimental host MR-signature) and activation of MR-proteins governing innate immune response programs (beneficial MR-signature). To identify MR-inverting compounds capable of rescuing activity of inactivated host MR-proteins, with-out adversely affecting the beneficial MR-signature, we developed the ViroTreat algorithm. Overall, >80% of drugs predicted to be effective by this methodology induced significant reduction of SARS-CoV-2 infection, without affecting cell viability. ViroTreat is fully generalizable and can be extended to identify drugs targeting the host cell-based MR signatures induced by virtually any pathogen.

The Host Cell ViroCheckpoint: Identification and Pharmacologic Targeting of Novel Mechanistic Determinants of Coronavirus-Mediated Hijacked Cell States

Most antiviral agents are designed to target virus-specific proteins and mechanisms rather than the host cell proteins that are critically dysregulated following virus-mediated reprogramming of the host cell transcriptional state. To overcome these limitations, we propose that elucidation and pharmacologic targeting of host cell Master Regulator proteins--whose aberrant activities govern the reprogramed state of infected-coronavirus cells--presents unique opportunities to develop novel mechanism-based therapeutic approaches to antiviral therapy, either as monotherapy or as a complement to established treatments. Specifically, we propose that a small module of host cell Master Regulator proteins (ViroCheckpoint) is hijacked by the virus to support its efficient replication and release. Conventional methodologies are not well suited to elucidate these potentially targetable proteins. By using the VIPER network-based algorithm, we successfully interrogated 12h, 24h, and 48h signatures from Calu-3 lung adenocarcinoma cells infected with SARS-CoV, to elucidate the time-dependent reprogramming of host cells and associated Master Regulator proteins. We used the NYS CLIA-certified Darwin OncoTreat algorithm, with an existing database of RNASeq profiles following cell perturbation with 133 FDA-approved and 195 late-stage experimental compounds, to identify drugs capable of virtually abrogating the virus-induced Master Regulator signature. This approach to drug prioritization and repurposing can be trivially extended to other viral pathogens, including SARS-CoV-2, as soon as the relevant infection signature becomes available.

Research capacity training for graduate students in basic medical sciences with the guideline of translational medicine / 中华医学科研管理杂志

In China,few research findings in basic medicine could be translated to clinical practice,which is known as the disconnection between basic research and clinical application.Such phenomenon is mainly due to lack of ability in translational medicine,particularly,lack of training in clinical research and translational medicine.Now the emergence of translational medicine has put forward new requirements for graduate education in medical school,and demanding that the patient-centered concept be strengthened.As the major force of scientific and technological innovation,graduate students should receive long-term systematic trainings and conduct original researches.In addition,advanced courses in clinical research and translational medicine should be offered to graduate students ;furthermore,translation research platforms should be built up for them so as to improve the capacity of scientific research and innovation.

Fasudil relives cerebral vasospasm following subarachnoid hemorrhage by inhibiting TLR 4 expression / 重庆医学

Objective To study the effect of Toll‐like receptor 4 inhibitor Fasudil on cerebral vasospasm after SAH .Methods A total of 60 New Zealand rabbits were randomly divided into three groups ,20 rabbits for each group ,SAH group:SAH model was established by autologous blood injection into the cisterna magna .Control group:saline was injected into cisterna magna .Fasudil group:Fasudil was injected into vein after SAH model .Vasospasm was valued by DSA and TCD .Seven days after operation basilar artery were collected .HE stain was used to observe vasospasm .TLR4 were observed by immunohistochemistry and western blot‐ting .Results Vasospasm model after SAH was successfully established .The basilar artery diameters were significantly shorter in the model group compared with the normal group (P<0 .01) .The artery diameter in Fasudil group increased significantly compared with SAH group(P<0 .01) .The expression of TLR4 decreased significantly in the Fasudil group compared with the model group (P<0 .05) .Conclusion Toll‐like receptor 4 pathway may be associated with cerebral vasospasm (DCV) .Fasudil could inhibit TLR‐4 expression and prevent cerebral vasospasm following subarachnoid hemorrhage .

Endovascular treatment of unruptured A1 segment aneurysms.

Aneurysms of the A1 segment of the anterior cerebral artery (A1A) are rare and challenging to treat. Less information is available regarding their management by endovascular approach. We evaluated our experience of endovascular treatment in 15 patients with unruptured A1As. We retrospectively reviewed unruptured A1As treated by embolization at our hospital. The clinical data and angiographic results were reviewed. A special technique involving shaping microcatheter tips was used for catheterization. From September 2009 to December 2012, 15 patients presenting with unruptured A1As were identified. All the patients were treated by selective embolization including five patients with balloon-assisted coiling (BAC) or stent-assisted coiling (SAC). These adjunctive techniques were used to catheterize the sac safely or to protect a branch at the neck. According to the location and direction of the aneurysm, "Z-shaped", "S-Shaped" or "U-Shaped" microcatheter tip shaping was used for microcatheter positioning and stabilization. All patients showed an excellent clinical outcome. A complete aneurysm occlusion was obtained in all the patients. Endovascular treatment of A1As is feasible and associated with good results. Because of their location and close relationship with perforators, endovascular treatment of A1As sometimes requires the use of BAC or SAC. The microcatheter tip shaping technique is very important for coiling. Our results suggest that endovascular treatment is a suitable therapeutic option for unruptured A1As when the aneurysm size is optimal for embolization.

Establishing selective course of English for international academic conference in medical university: a case study / 中华医学教育探索杂志

In order to explore the implement of English for academic purposes (EAP) teaching,the selective course of English for international academic conference was opened up in the Third Military Medical University.The teaching practice was enacted based on 50 selected medical undergraduates,with the feature of modularized teaching,experimental study and teaching team.Teaching outcome was assessed by means of students' record,investigation questionnaire,teaching video and teachers' journal,which indicated that the course of English for international academic conference played the positive role in promoting student's ability for EAP and the key questions included how to make it interesting while keeping it professional and the quality of teachers.

Direct protection of cultured neurons from ischemia-like injury by minocycline / 대한해부학회지

Minocycline, a tetracycline antibiotic, is now known to protect cells via an anti-inflammatory mechanism. We further explored this effect using an in vitro model of ischemia-like injury to neurons. Coculturing neurons with microglia, the brain's resident immune cell, modestly increased cell death due to oxygen and glucose deprivation (OGD), compared to neurons alone. Treatment of cocultures with minocycline decreased cell death to a level significantly lower than that of neurons alone. Treatment of cocultures with minocycline or inhibitors of various immune mediators, also led to decreased cell death. Importantly, treatment of neuron cultures without added microglia with these same inhibitors of tissue plasminogen activator, matrix metalloproteinases, TNF-alpha and inducible nitric oxide synthase as well as minocycline also led to decreased cell death. Thus, anti-inflammatory treatments appear to be directly protective of neurons from in vitro ischemia.

Expression level of Interferon regulatory factor 5 in systemic lupus erythematosus / 中华风湿病学杂志

Objective To compare the expression level of interferon regulator factor 5 (IRF5) of the health controls and systemic lupus erythematosus (SLE) patients and analyze the relationship between IRF5 and SLE disease activity. Methods Peripheral blood monoeytes (PBMCs) from SLE patients and healthy donors were separated with Ficoll density gradient eentrifugation and total cellular RNA was isolated with Trizol from the PBMCs, the mRNA was reverse transcripted to cDNA. Real-time PCR was applied to determine the expression level of IRFS. The expression level of IRF5 between the two groups were compared. The correlations of expression level of IRF5 with SLE disease activity and other laboratory or clinical parameters of SLE patients were analyzed. Results The level of IRF5 was (2.1±2.2) in SLE patients and (1.5±1.2) in healthy controls, the difference was not statistically significant (P=0.161). And the levels of IRF5 in SLE patients were significantly correlated with their SLEDAI (r=0.616,P<0.01), but not correlated with other parameters such as bemoglobulin complements, immunoglobulin etc. Anti-dsDNA antibody positive patients had significantly higher expression of IRF5 compared to the anti-dsDNA-antibedy-negative patients. The IRF5 mRNA levels of SLE patients with fever or neuropsyehiatric symptoms were significantly higher than those of patients free of neuropsychiatrie involvement. Conclusion High expression level of 1RF5 may contribute to the pathogenesis of SLE in disease activity and antibody production.

The significance of anti-p68 antibody in rheumatoid arthritis / 中华风湿病学杂志

Objective To evaluate the diagnostic specificity and sensitivity of anti-p68 antibody in rheumatoid arthritis (RA), and to analyze the relationship between anti-p68 antibody and the activity of RA . Methods Anti-p68 antibody was tested in the serum of 183 RA patients and 195 controls by indirect immunofluorescence (IIF). The clinical characteristics of RA between anti-p68 positive group and negative group were compared. Results ① The positive rate of anti-p68 antibody was 67.8% (124/183) in RA, 14.9% (17/114) in other rheumatic diseases and none in normal controls (0/81). Anti-p68 antibody was different from other auto-antibodies found in RA. ② Its sensitivity was 67.8% and specificity was 91.3% for RA, positive prediction rate was 87.9% and negative prediction rate was 75.1%. ③ There was statistic difference between the positive group and negative group in disease duration, RF and degree of X-ray changes. Conclusion ① Anti-p68 antibody is a new and specific serum maker for diagnosis of RA. ② The antibody is correlated with disease duration, RF and X-ray changes.

Diagnostic significance of combined detection of rheumatoid factor and anticyclic citrullinated peptide antibodies in rheumatoid arthritis / 北京大学学报(医学版)

Objective:To evaluate the significance of the combination of rheumatoid factor (RF) and anti-filaggrin antibodies (AFAs) in the diagnosis of rheumatoid arthritis (RA). Methods:Sera from 266 RA patients and 186 controls were studied. RF and some kinds of AFAs (including antikeratin antibodies, anti-perinuclear factor and anticyclic citrullinated peptide antibodies) were detected using immunofluorescence and ELISA respectively. The distributions of these antibodies were compared to determine the significance of the combination of RF and AFAs. Results:The sensitivity and secificity of RF for the diagnosis of RA were 65.8% and 81.8% respectively, as compared with those of anti-CCP or RF positive were 86.5% and 80.6% respectively. Only a slight change was found in the secificity and negative predictive value between them. RF could not be detected in 91 patients while 64.8% of them could be found to have at least one kind of AFAs. Conclusion:AFAs, particular anticyclic citrullinated peptide antibodies, could be helpful for the diagnosis of RA especially in RF-negative patients.

Immunological characteristics of chronic graft versus host diseases murine model with lupus nephritis / 北京大学学报(医学版)

Objective:To establish the murine systemic lupus erythematosus (SLE) model of chronic graft versus host diseases(cGVHD). To analyze the pathological changes and serological and immunological features in the animals. Methods: Female (C57BL/10?DBA/2)F1 hybrids aged 6-8 weeks were randomly divided into model group and healthy controls (HC). Lymphocytes from female DBA/2 were injected intravenously to the model group on days 0, 3 and 8,while PBS were injected to the HC under the same condition as a control group. Bradford was applied to monitor the development of albuminuria quantitively. Sera were tested by enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF) for the presence of autoantibodies. To compare the differences of CD4+CD25+ Treg cells between the two groups by flow cytometry (FCM) and the differences in the expression of Foxp3 by real time polymerase chain reaction(RT-PCR). The kidneys of model mice were removed in the 12th week and were made frozen sections for direct immunofluorescence(DIF)and paraffin imbedding for PASM staining. Results: The titers of proteinuria in model group in the 6th week, 8th week, 10th week, and 12th week were significantly higher than those of the HC groups(P=0.004, 0.005,respectively). The titers of anti-dsDNA and anti-nucleosome antibodies were significantly increased in the model group compared with the HC (P

Anemia in patients with rheumatoid arthritis / 中国实用内科杂志

Objective To investigate the prevalence of anemia in patients with rheumatoid arthritis (RA) and the relationship between anemia and disease activity of RA. Methods Retrospectively analyze the data of 239 RA patients. Results (1)Estimates of the prevalence of the anemia was 51.4% ;the level of anemia was mild and moderate,and none was severe. (2)There were statistic differences between the patients with anemia and without anemia in disease duration,morning stiff ness,ESR,CRP and degree of X-ray change. (3)There were statistic differences between the patients with anemia and without anemia in leukocyte counts and platelet counts. (4)The level of hemoglobin (Hb) was significantly increased after anti-rheumatoid treatment. Conclusion (1)Anemia is a common sign in individuals with rheumatoid arthritis. The level of anemia is mild and moderate. (2)There are important associations between anemia and disease activity of RA. (3)The level of hemoglobin is increased after anti-rheumatoid treatment.

Expression of vascular endothelial growth factor in squamous cell carcinoma of oral cavity / 实用口腔医学杂志

Objective: To study the role of vascular endothelial growth factor (VEGF) during angiogenesis in oral squamous cell carcinoma (SCC).Methods:Surgical specimens from 45 patients with oral SCC were examined for VEGF expression and microvessel density(MVD) by immunocytochemical staining. Anti-VEGF polyclonal antibody was used to determine VEGF expression and anti-CD 34 monoclonal antibody was used to determine MVD. Results:①VEGF protein was mainly found in cytoplasm of cancer cells and in few endothelial cells. Stained microvessels were mainly distributed in the regions of the connective tissue near the cancer nest;②there was a close positive correlation between MVD and VEGF expression;③SCC with high MVD or high VEGF level was mostly found with metastasis in cervical lymph node (P