System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has become a global crisis. To gain systems-level insights into its pathogenesis, we compared the blood proteome and phosphoproteome of ICU patients with or without SARS-CoV-2 infection, and healthy control subjects by quantitative mass spectrometry. We find that COVID-19 is marked with hyperactive T cell and B cell signaling, compromised innate immune response, and dysregulated inflammation, coagulation, metabolism, RNA splicing, transcription and translation pathways. SARS-CoV-2 infection causes global reprogramming of the kinome and kinase-substrate network, resulting in defective antiviral defense via the CK2-OPN-IL-12/IFN-/{beta} axis, lymphocyte cell death via aberrant JAK/STAT signaling, and inactivation of innate immune cells via inhibitory SIRPA, SIGLEC and SLAM family receptor signaling. Our work identifies CK2, SYK, JAK3, TYK2 and IL-12 as potential targets for immunomodulatory treatment of severe COVID-19 and provides a valuable approach and resource for deciphering the mechanism of pathogen-host interactions.

Epitope-resolved serology test differentiates the clinical outcome of COVID-19 and identifies defects in antibody response in SARS-CoV-2 variants

BACKGROUNDThe role of humoral immunity in the coronavirus disease 2019 (COVID-19) is not fully understood owing, in large part, to the complexity of antibodies produced in response to the SARS-CoV-2 infection. There is a pressing need for serology tests to assess patient-specific antibody response and predict clinical outcome. METHODSUsing SARS-CoV-2 proteome and peptide microarrays, we screened 146 COVID-19 patients plasma samples to identify antigens and epitopes. This enabled us to develop a master epitope array and an epitope-specific agglutination assay to gauge antibody responses systematically and with high resolution. RESULTSWe identified 54 linear epitopes from the Spike (S) and Nucleocapsid (N) protein and showed that epitopes enabled higher resolution antibody profiling than protein antigens. Specifically, we found that antibody responses to the S(811-825), S(881-895) and N(156-170) epitopes negatively or positively correlated with clinical severity or patient survival. Moreover, we found that the P681H and S235F mutations associated with the coronavirus variant B.1.1.7 altered the specificity of the corresponding epitopes. CONCLUSIONSEpitope-resolved antibody testing not only offers a high-resolution alternative to conventional immunoassays to delineate the complex humoral immunity to SARS-CoV-2 and differentiate between neutralizing and non-neutralizing antibodies, it may also be used as predictor of clinical outcome. The epitope peptides can be readily modified to detect antibodies against variants in both the peptide array and latex agglutination formats. FUNDINGOntario Research Fund (ORF)-COVID-19 Rapid Research Fund, the Toronto COVID-19 Action Fund, Western University, the Lawson Health Research Institute, the London Health Sciences Foundation, and the AMOSO Innovation Fund.

High-molecular-weight hyaluronan produced by activated pancreatic stellate cells promotes pancreatic cancer cell migration via paracrine signaling.

BACKGROUND: The polysaccharide hyaluronan (HA) is abundant in pancreatic cancer (PC) tissue and promotes pancreatic cancer cell (PCC) motility in vitro. However, it is controversial as to whether high-molecular-weight HA (HMW-HA) or low-molecular weight HA(LMW-HA) is present in the pancreatic cancer stroma and whether PCC or pancreatic stellate cell (PSC) in PC tissue produces HA. We thereby aim to characterize the molecular weight and source of HA in PC tissue that promotes cancer cell motility. METHODS: We analyzed the expression of hyaluronan synthase 2 (HAS2) and the hydrolyzing enzyme hyaluronidase 1 (HYAL1) in PCC lines and pancreatic stellate cells (PSCs) using real-time PCR. HA production in the supernatant of PCC lines and PSCs and in PC tissues was quantitatively and qualitatively examined. Finally, we knocked down HYAL1 expression in one of the PCC line PANC-1 cells and analyzed the impact on cell migration. RESULTS: HAS2 was abundantly expressed in activated PSCs (aPSCs) but less so in quiescent PSCs (qPSCs) and PCC lines. The baseline expression of HYAL1 did not differ among the cell types. The concentration of HMW-HA was higher in the supernatant of aPSCs than in that of PCC lines. Treatment with exogenous HMW-HA promoted PANC-1 cell motility. Knockdown of HYAL1 decreased HMW-HA-promoted PANC-1 cell migration, which was accompanied by a decrease in intracellular HA levels. CONCLUSION: aPSCs are an important source of stromal HMW-HA, which promotes PCC migration in an HYAL1-dependent manner in PC.

Effects of moxibustion at different time points on foot swelling, serum level of TNF-α and circadian rhythm in rats with rheumatoid arthritis / 中国针灸

OBJECTIVE@#To study the effects of moxibustion at different time points on serum level of tumor necrosis factor-α (TNF-α) in rats with rheumatoid arthritis (RA), and to explore its regulation mechanism on circadian rhythm.@*METHODS@#A total of 96 Sprague-Dawley (SD) adult rats were randomly assigned into a blank group, a model group, a moxibustion at 5-7 AM group and a moxibustion at 5-7 PM group, 24 rats in each group, half male and half female. Each group was further divided into a 0 AM group, a 6 AM group, a 12 N group and a 6 PM group, 6 rats in each group. All rats were treated with the 12 h/12 h light-dark cycle in the whole process of experiment. Except for the blank group, all rats were treated with intracutaneous injection of freund's complete adjuvant (FCA) at right foot pad to establish the RA model. The rats at the two moxibustion groups were treated with grain-sized moxibustion at "Shenshu" (BL 23) and "Zusanli" (ST 36) at 5-7 AM and 5-7 PM, respectively, one side per treatment, once a day; six treatments were taken as one course and 3 courses were given with an interval of one day between courses. The rats in the remaining groups were treated with identical fixation but without moxibustion intervention. The right foot volume was measured before model establishment, after model establishment and after treatment. The blood samples were collected after treatment and the serum level of TNF-α was measured by ELISA. The SPSS 21.0 software and Halberg Cosinor were adopted to analyze the experiment data.@*RESULTS@#After treatment, compared with the blank group, the foot swelling severity was significantly increased in the model group, moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (all <0.01); compared with the model group, the foot swelling severity was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.01). Compared with the blank group, the serum level of TNF-α was increased significantly in the model group and moxibustion at 5-7 AM group (both <0.05); compared with the model group, the serum level of TNF-α was reduced significantly in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (both <0.05). The serum level of TNF-α showed circadian rhythm in all the groups (all <0.05), and the peak appeared at night phase; compared with the blank group, the median value of TNF-α was increased significantly in the model group (<0.05), the peak phase was delayed and the amplitude was increased (<0.05); compared with the model group, the median value of TNF-α was significantly reduced in the moxibustion at 5-7 AM group and moxibustion at 5-7 PM group (<0.01), the peak phase was advanced and the amplitude was reduced (<0.05).@*CONCLUSION@#Moxibustion could effectively reduce the serum level of TNF-α to relieve the foot swelling severity in RA rats. Moxibustion could regulate the circadian rhythm of TNF-α to play its effects on the inhibition of the synthesis of TNF-α. No efficacy is observed between the treatment at 5-7 AM and 5-7 PM.

Effects of grain-sized moxibustion from 7 am to 9 am on circadian rhythm of inflammatory factor IL-6 in rats with rheumatoid arthritis / 中国针灸

<p><b>OBJECTIVE</b>To explore the rhythm regulatory mechanism of interleukin-6 (IL-6) in the process of moxibustion for rheumatoid arthritis (RA).</p><p><b>METHODS</b>A total of 144 Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, a moxibustion group, a sham operation group, an operation group, an operation+moxibustion group, 24 rats in each one. Each group was divided into 4 time points (0:00 am, 6:00' am, 12:00 am, 6:00 pm), 6 rats in each time point. The Light-Dark 12 : 12 was given in all rats for light-dark cycle. Except the blank group, rats in the remaining groups were treated with intracutaneous injection of freund's complete adjuvant at right-side foot to establish the model of RA. After the model establishment, bilateral adrenal, glands were removed in the operation group and operation + moxibustion group, while those in the sham operation group were not removed with identical operation procedure. Rats in the moxibustion group and operation + moxibustion group were treated with grain-sized moxibustion from 7:00 am to 9:00 am at "Shenshu" (BL 23) and "Zusanli" (ST 36) once everyday, 6 times were taken as one session and 3 sessions were required tatclly, while rats in the remaining groups received identical fixation without moxibustion. The general health state and foot volume of rats were measured before model establishment, after establishment and after treatment. After treatment, rats were sacrificed at each time point to collect the blood sample and measure the content of IL-6 by using enzymne-immunoassay method.</p><p><b>RESULTS</b>Compared with the blank group, the foot swelling in the model group was obviously increased (P<0. 05); the IL-6 maintained circadian rhythm (P<0. 05), but the peak phase had a backward trend, famplitude had an increased trend and the median was significantly lifted (P<0. 05). Compared with model group, !the foot swelling in the moxibustion group was obviously decreased (P<0. 05); the IL-6 maintained circadian. rhythm (P<0. 05), and the peak phase had a forward trend, amplitude had a decreased trend and the median was significantly reduced (P<0. 05). Compared with the moxibustion group, the foot swelling in the operation--moxibustion group was obviously increased (P < 0.05); the IL-6 maintained circadian rhythm (P < 0.5), but the peak phase moved forwrd, and the median was significantly elevated (P < 0.05).</p><p><b>CONCLUSION</b>The IL-6 in plasma maintains significant pathological circadian rhythm in RA rats; with the complete hypothalamic-pituitary-adrenal axis, moxibustion is likely to regulate the circadian rhythm of IL-6 to play an important role of anti-inflammatory effect in RA rats.</p>

Effects of deep brain stimulation of bilateral nucleus accumbens on the behavior of relapse in morphine-dependent rats / 中华行为医学与脑科学杂志

Objective To investigate the influence on the behavior of withdrawal and relapse after deep brain stimulation of bilateral nucleus accumbens in morphine-dependent rats. Methods The rats with a strong unconditioned preference were discarded in preconditioning test, the selected rats were distributed into five groups randomly. After operation,morphine hydrochloride was injected subcutaneously into SD rats for 12 days (once every day,initial 5 mg/kg,increasing by 5 mg/kg per time,stable in 20 mg/kg ). A modified electrical circuit was used to procedure the DBS,the parameter was 130 Hz,150 A,60 s,l h/d,14 d. CPP test was used to exam the effect of DBS. A minor morphine dose (3 mg/kg) was injected to induce the behavior of relapse, and CPP was tested again after 24 h. Two-way ANOVA was performed on the data with Bonferroni posttest. Result ①After CPP training,CPP score of group morphine, morphine + sham and morphine + DBS was ( 155. 87 ± 20. 45 ) s, (107.33 ± 18.10)s,(135.45 ±22.09)s,and had significant difference with group of control( ( -70.34 ± 15.40) s)(t = 9.45,P＜0.01; t = 6.94,P＜0.01;t = 8.04,P＜0.01).②After 7 days' DBS,the CPP score in group of morphine + DBS reduced significantly compared to group of morphine( t = 4.21, P＜0.01) and morphine + sham( t=1.10, P＜0.05).0n the 14th day,there was more pronounced reduction ( t = 5. 15, P＜0.01; t = 3.92, P＜ 0.01). ③ 24 hours after the minor morphine dose was injected,the CPP score in morphine + DBS didn't increase significantly, and had significant difference with group of morphine ( t = 4.04, P＜0.01) and morphine + sham ( t= 4. 13, P＜0.01). Conclusion DBS bilateral nucleus accumbens in morphine-dependent rats can interfere the behavior of morphine-induced CPP and relapse.

Quality Control of the Research on Mechanisms of Acupuncture Therapy by Using PET-CT Imaging Techniques / 针刺研究

Combining the functional metabolic imaging and anatomical imaging,PET-CT stands for the highest level of current nuclear medical imaging techniques,and has been applying increasingly in acupuncture studies.However,owning to complexity of the brain function and sensitivity of brain metabolism,the lower reproducibility of cerebral function imaging even may reverse many results.This has been provoked more and more attention by researchers.Its main cause is intimately related to poor experimental methodology.For this reason,the present paper raises the quality control of the research on mechanisms of acupuncture in the process of application of PET-CT techniques from 1) the included standards of participants,2) the preparation of participants during the scanning process of PET-CT,3) the setting of the scanning parameters,4) the used machine and the imaging reagent,5) the analysis of dada,6) the standardization of acupuncture manipulation,and 7) the designs of the acupuncture operation,needling opportunity and scanning opportunity,hoping to offer some beneficial information for the coming researches.

XPS and XRD study of FeCl3-graphite intercalation compounds prepared by arc discharge in aqueous solution.

A novel one-step synthesis method of FeCl3-graphite intercalation compounds (FeCl3-GICs) by an arc discharge in aqueous solution was reported for the first time. It presented a simply and controllable way to synthesize FeCl3-GICs. The structure of the stage 7 GICs was examined and characterized by X-ray diffraction. X-ray photoelectron spectroscopic study of stage 7 of FeCl3-GICs was also carried out. The change in the binding energy suggests the nature of charge transfer and lowering of Fermi level as has been reported previously for other acceptor intercalation compounds.

Study on the heat resistant property of Ag/4A antibacterial agent.

The heat resistant property of silver-loading zeolite 4A antibacterial agent (SLZ) was investigated by heat treatment methodology. The morphological and structural changes of the specimens were characterized by using differential thermal analyzer (TG-DTA), scanning electron microscopy, and X-ray diffraction techniques. The particle size of the specimens was measured by Malvern instruments zetasizer systems, and silver content of specimens was determined by inductively coupled plasma spectrometer. Escherichia coli (E. coli) was chosen as indicators of fecal contamination to evaluate the antibacterial effect of the specimens by minimum inhibitory concentration method. The service life of the specimens was also tested. The experimental results indicated that as heat-treatment temperature increases, the particle size increases, more aggregating occurs, silver content decreases, the color of SLZ gradually changes from white to brown, and the antibacterial ability falls. The release rate of Ag(+) cation from SLZ became slow after heat treatment at 400, 450, and 500 degrees C, thus resulting in prolonged service life of SLZ. When the heat-treatment temperature approached 800 degrees C, the collapse of SLZ structure occurred, and the crystals of SiO(2) and Al(2)O(3) were formed after recrystallization. Consequently, the heat resistant temperature of SLZ can be as high as 500 degrees C. SLZ could be used in antibacterial plastics, antibacterial fibers, or biomedical materials.

Influences of transforming growth factor-?1 on scavenger receptor class A and class B (CD36) in THP-1 derived macrophages / 吉林大学学报(医学版)

Objective To investigate the influence of transforming growth factor-?1(TGF-?1) on macrophage scavenger receptor (ScR) class A and B(ScR-B,CD36) in order to provide theoretical foundation for expounding the formation and therapy of atherosclerosis(AS).Methods THP-1 derived macrophages were divided into control group and experimental group,the cells in experimental group were treated with 3.0 mg?L-1Anti TGF-?1 Ab,the cells in control group were treated with 3.0 mg?L-1 IgG. The 125I-acetylated low density lipoprotein (125I-Ac-LDL for ScR-A) and 125I-oxidized low density lipoprotein (125I-Ox-LDL for CD36) uptaking (including 4℃ binding,37℃ association and degradation) were measured respectively.Influence of anti TGF-?1 antibody (Anti TGF-?1 Ab) on the ScR-A and CD36 mRNA expressions in THP-1 derived macrophages was measured meanwhile,respectively.Results Compared with control group ( binding: 8.23 ?g?g-1 ?1.24 ?g?g-1 protein,association:45.69 ?g?g-1 ?6.92 ?g?g-1 protein,and degradation: 112.18 ?g?g-1 ?20.15 ?g?g-1 protein),Anti TGF-?1 Ab increased 125I-Ac-LDL binding(48.67 ?g?g-1 ?6.52 ?g?g-1 protein),association (412.30 ?g?g-1?12.21 ?g?g-1 protein),and degradation (896.48 ?g?g-1 ?32.74 ?g?g-1 protein) significantly (P

EFFECTS OF GINSENG ROOT SAPONINS ON IMMUNOSUPPRESSION IN TUMOR-BEARING MICE / 中国药理学通报

Ginseng root saponins ( GRS ) was extracted from native ginseng.The dosage used was calculated according to the content of saponins.Immunosuppression in mice was induced in 15 days after implar- ting Ehrlich ascites carcinoma (EAC) subcutaneously in armpit, and in 5-11 days after implanting EAC into peritoneal cavity. GRS administered orally 50mg/kg- for 8 days did not prevent the atrophy of the thymus in tumor-bearing mice but rather aggravated it. GRS administered orally 50mg/kg for 14 days partially restored the suppressed phagocytosis of peritoneal macrophages in tumor-bearing mice. GRS administered orally 50mg/kg for 6 days somewhat restored the suppression of hemolysin formation in tumor-bearing mice 5 days after implanting EAC, But GRS given in the same route and dosage for 9 days, showed no effect on the suppression of hemolysin formation. GRS administered orally 50mg/kg for 10 days partially restored the suppression of delayed hypersensitive reaction in tumor-bearing mice.