Successful treatment with plasma exchange followed by intravenous immunoglobulin in a critically ill patient with COVID-19.

Here we report a case of a laboratory-confirmed 2019 novel coronavirus (2019-nCoV)-infected patient with COVID-19 (coronavirus disease 2019) who developed respiratory failure and shock accompanied by persistent diarrhoea despite conventional therapeutic interventions. The patient avoided mechanical ventilation and showed an immediate clinical and radiological improvement following treatment with intensive plasma exchange (PE) followed by intravenous immunoglobulin (IVIG). Successful therapeutic strategies in this case suggest that timely initiation of PE treatment followed by IVIG in critically ill patients with COVID-19 may prevent the disease from worsening and help to reduce the requirement for mechanical ventilation and intensive supportive care. Moreover, it may improve poor clinical outcomes of these patients.

[The effects of dendritic cells overexpressing Serrate1 on the differentiation of CD4(+)CD25(+) T cells in asthmatic mice].

OBJECTIVE: To observe the effects of dendritic cells (DCs) overexpressing Serrate1 on the differentiation of CD(4)(+)CD(25)(+) T cells and the production of inhibitory cytokines in asthmatic mice. METHODS: Asthma mouse model was established by the routine method. After intravenous injection of the DCs transfected with Serrate1 into the naïve mice, the airway and lung inflammation was observed and the count of CD(4)(+)CD(25)(+) T cells delivered from the spleen and the percentage accounting for CD(4)(+)T cells were measured by flow cytometry. The expression of IL-10, transforming growth factor (TGF)beta1, and cytologic T-lymphocyte-associated antigen (CTLA)-4 mRNA of CD(4)(+) T cells were detected by RT-PCR. RESULTS: Compared with the non-transfected group, the airway inflammation of asthmatic mice injected the DCs transfected with Serrate1 reduced significantly, the number of CD(4)(+)CD(25)(+) T cells delivered from the spleen and the percentage accounting for CD(4)(+) T cells and the expression of IL-10, TGFbeta1, and CTLA-4 mRNA of CD(4)(+) T increased. CONCLUSIONS: DCs overexpressing Serrate1 could induce the differentiation of the CD(4)(+)CD(25)(+) T cells and increase the production of inhibitory cytokines in asthma in vivo, indicating that Notch1/Serrate1 signal pathway of DCs plays an important role in the induction of immune tolerance of T cells to allergens.

Efficacy of Shenqi Pollen Capsules for High-Altitude Deacclimatization Syndrome via Suppression of the Reoxygenation Injury and Inflammatory Response.

High-altitude deacclimatization syndrome (HADAS) is involved in hypoxia-reoxygenation injury and inflammatory response, induced a series of symptoms, and has emerged as a severe public health issue. Here, we investigated the mechanism as well as potential means to prevent HADAS using Shenqi pollen capsules (SPCs) in subjects with HADAS in a multicenter, double-blinded, randomized, placebo-controlled study. All subjects were at the same high altitude (3650 m) for 4-8 months before returning to lower altitudes. Subjects (n = 288) in 20 clusters were diagnosed with mild or moderate HADAS on the third day of the study. We randomly allocated 20 clusters of subjects (1 : 1) to receive SPCs or a placebo for 7 weeks, and they were then followed up to the 14th week. The primary endpoints were subjects' HADAS scores recorded during the 14 weeks of follow-up. Compared with the placebo, SPC treatment significantly decreased the subjects' HADAS scores and reduced the incidence of symptom persistence. SPC therapy also reduced the serum levels of CK, CK-MB, LDH, IL-17A, TNF-α, and miR-155 and elevated IL-10 and miR-21 levels. We thus demonstrate that SPCs effectively ameliorated HADAS symptoms in these subjects via suppression of the hypoxia-reoxygenation injury and inflammatory response.

Identification of Streptococcus mitis321A vaccine antigens based on reverse vaccinology.

Streptococcus mitis (S. mitis) may transform into highly pathogenic bacteria. The aim of the present study was to identify potential antigen targets for designing an effective vaccine against the pathogenic S. mitis321A. The genome of S. mitis321A was sequenced using an Illumina Hiseq2000 instrument. Subsequently, Glimmer 3.02 and Tandem Repeat Finder (TRF) 4.04 were used to predict genes and tandem repeats, respectively, with DNA sequence function analysis using the Basic Local Alignment Search Tool (BLAST) in the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Cluster of Orthologous Groups of proteins (COG) databases. Putative gene antigen candidates were screened with BLAST ahead of phylogenetic tree analysis. The DNA sequence assembly size was 2,110,680 bp with 40.12% GC, 6 scaffolds and 9 contig. Consequently, 1,944 genes were predicted, and 119 TRF, 56 microsatellite DNA, 10 minisatellite DNA and 154 transposons were acquired. The predicted genes were associated with various pathways and functions concerning membrane transport and energy metabolism. Multiple putative genes encoding surface proteins, secreted proteins and virulence factors, as well as essential genes were determined. The majority of essential genes belonged to a phylogenetic lineage, while 321AGL000129 and 321AGL000299 were on the same branch. The current study provided useful information regarding the biological function of the S. mitis321A genome and recommends putative antigen candidates for developing a potent vaccine against S. mitis.

[Effects and mechanism of CD4+ CD25+ T cells on the airway inflammation of asthmatic mice].

OBJECTIVE: To investigate the effects and mechanism of CD(4)(+) CD(25)(+) T cells on the airway inflammation of asthmatic mice. METHODS: Sixty mice were divided into 3 groups (20 in each group) including group A [ovalbumin (OVA) sensitized/challenged mice], group B (saline sensitized/challenged mice), group C [CD(25)(+) T cells deleted by anti-CD(25)(+) and OVA sensitized/challenged mice]. Mice of A group were sensitized on days 1 and 13 by OVA and challenged from days 21 to 29 by 2% OVA 10 ml repeatedly to establish a murine model of asthma characterized by airway inflammation. The mice of group B were treated by saline 10 ml. The mice of group C were treated by the same method of group A after deletion of the CD(25)(+) T cells by anti-mouse CD(25)(+) monoclonal antibody (McAb). Lymphocytes were separated from the spleen of the three groups and the number of T cells were calculated by Fluorescence Activated Cell Sorter (FACS). The CD(4)(+) T cells were purified and the expression of interleukin-10 (IL-10), transforming growth factor-beta(1) (TGF-beta(1)) and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) mRNA were analyzed by the method of reverse transcriptase-polymerase chain reaction (RT-PCR). The airway inflammatory indices were detected by hematoxylin and eosin (HE) staining. RESULTS: In repeatedly OVA-challenged mice (group A), the ratio of CD(4)(+) CD(25)(+) T cells to CD(4)(+) T cells decreased significantly [(3.10 +/- 0.03)%] as compared with group B [(9.60 +/- 0.04)%, P < 0.01]. The expression of IL-10, CTLA-4 mRNA and TGF-beta(1) mRNA of OVA-challenged mice (group A) decreased significantly (0.250 +/- 0.040 vs 0.29 +/- 0.03, 0.28 +/- 0.06 vs 0.480 +/- 0.080, 0.47 +/- 0.05 vs 0.50 +/- 0.03, all P < 0.01). The expression of IL-10, CTLA-4 mRNA and TGF-beta(1) mRNA of the mice treated by anti-CD(25)(+) McAb (group C) decreased as compared with groups A and B (0.080 +/- 0.020, 0.11 +/- 0.04, 0.12 +/- 0.05, P < 0.01). The inflammation in the lungs of mice with depleted CD(4)(+) CD(25)(+) T cells by anti-CD(25)(+) McAb was more marked than that in the OVA-challenged and control group mice. CONCLUSION: The polarization of CD(4)(+) CD(25)(+) T cells in asthmatic mice decreased significantly, which may play an important role in the pathogenesis of asthma.

Berberine attenuates cigarette smoke-induced acute lung inflammation.

Berberine (Ber), the major constituent of Coptidis Rhizoma, possesses anti-inflammatory properties. In this study, we investigated the effects of Ber on cigarette smoke (CS)-mediated acute lung inflammation. C57BL/6 mice (6-8 weeks) were exposed to CS to induce acute lung injury. Ber was used to pretreat CS-exposed mice (50 mg/kg, intragastrically). Lung tissues were collected for histological examination, myeloperoxidase (MPO) activity assay, Western blot analysis, and electrophoretic mobility shift assay. Bronchoalveolar lavage fluid (BALF) was measured for cell counts and cytokine analysis. Histological examination showed that CS exposure caused infiltration of inflammatory cells into alveolar spaces and interstitial edema. Pretreatment with Ber significantly attenuated CS-induced lung inflammation. The numbers of total cells, macrophages, and neutrophils in BALF were decreased by 43, 40, and 53 %, respectively, by Ber pretreatment in CS-exposed mice, accompanied by decreased MPO activity, a marker of neutrophil accumulation. Ber pretreatment also profoundly diminished CS-induced secretions of macrophage inflammatory protein 2, tumor necrosis factor alpha, interleukin-6, and monocyte chemotactic protein-1 in BALF, along with less nuclear translocation of the pro-inflammatory transcription factor nuclear factor-kappa B (NF-κB) p65 subunit and lower NF-κB DNA-binding activity (P < 0.01). Thus, our results indicated that Ber ameliorates CS-induced acute lung injury through its anti-inflammatory activity.

Regulation of S100A4 expression via the JAK2-STAT3 pathway in rhomboid-phenotype pulmonary arterial smooth muscle cells exposure to hypoxia.

To investigate the effect of JAKs-STATs signal pathway on expression of S100A4 in pulmonary arterial smooth muscle cells (PASMCs), the action of S100A4 and hypoxia induced factor 1 (HIF-1) on the proliferation of hypoxic PASMCs. The results showed that S100A4 immunostaining was localized in the cytoplasm and nuclei of PASMCs exposure to hypoxia and it was predominantly expressed in rhomboid cells (R-SMCs). The mRNA and protein levels of S100A4 expression increased in PASMCs after hypoxic stimulus for 4, 8, 16 h. The immunofluorescence intensity and protein levels of S100A4 were suppressed, and the number of R-SMCs was reduced, when pretreatment with HIF-1α siRNA, STAT3 siRNA, S100A4 siRNA, and S100A4 inhibitor NSC 95397. Pretreatment with HIF-1α siRNA and anti-IL-6 antibodies, the levels of phospho-JAK2, -STAT3, and S100A4 were decreased, while HIF-1α kept stable in hypoxic cells. Importantly, pretreatment with HIF-1α siRNA, anti-IL-6 antibodies, STAT3 siRNA, and S100A4 siRNA, significantly attenuated the proliferation of PASMCs exposure to hypoxia. These data demonstrate that S100A4 is predominantly expressed in hypoxic R-SMCs, and regulated by the activation of JAK2-STAT3 signal pathway, which is dependent on hypoxia-induced HIF-1α expression. These results suggest that JAK2-STAT3 and HIF-1α could serve as targets for the regulation of phenotype modulation of PASMCs during the process of pulmonary vessel lesions.

Addition of theophylline or increasing the dose of inhaled corticosteroid in symptomatic asthma: a meta-analysis of randomized controlled trials.

PURPOSE: Low-dose theophylline has anti-inflammatory effects. The aim of this study was to evaluate the effects of adding theophylline compared with increasing the dose of inhaled corticosteroid (ICS) on symptomatic asthma. MATERIALS AND METHODS: The associated literature was acquired through deliberate searching and selected based on the established inclusion criteria for publications. The extracted data were further analyzed by a meta-analysis. RESULTS: Four randomized, controlled, parallel studies were selected. Addition of theophylline produced a greater increase of forced expiratory volume in one second as %predicted (FEV1pred) by 2.49% [95% confidence interval (CI) 1.99-3.00; z = 9.70; p < 0.001], compared with increasing the dose of ICS. There was no difference between the two treatments in terms of peak expiratory flow (PEF). CONCLUSION: Addition of theophylline to ICS has similar therapeutic effects on improving lung function as increasing the dose of ICS in the treatment of symptomatic asthma.

Rab1 GTPase promotes expression of beta-adrenergic receptors in rat pulmonary microvascular endothelial cells.

It is known that Rab1 regulates the expression and function of beta-adrenoceptors (beta-ARs) in many cells. However, the effect of these changes in rat pulmonary microvascular endothelial cells (RPMVECs) is not known. In the present study, we investigated the role of Rab1, a Ras-like GTPase that coordinates protein transport from the endoplasmic reticulum (ER) to the Golgi body and regulates the cell-surface targeting and function of endogenous beta-ARs in RPMVECs in the presence of lipopolysaccharide (LPS). We found that lentivirus-driven expression of wild-type Rab1 (Rab1WT) in RPMVECs strongly enhanced the amount of beta-ARs on the cell surface, whereas the dominant-negative mutant Rab1N124I significantly attenuated beta-ARs expression on the cell surface. In addition, LPS stimulation significantly reduced beta-ARs expression on the cell surface in RPMVECs; however, this effect was reversed by over-expression of wild-type Rab1WT. Fluorescent microscopy analysis demonstrated that expression of Rab1N124I and Rab1 small interfering RNA (siRNA) significantly induced the accumulation of green fluorescent protein (GFP)-tagged beta(2)-AR in the ER. Consistent with their effects on beta-ARs export, Rab1WT and Rab1N124I differentially modified the beta-AR-mediated activation of extracellular signal-regulated kinase1/2 (ERK1/2). Importantly, over-expression of Rab1WT markedly reduced LPS-induced hyper-permeability of RPMVECs by increasing the expression of beta(2)-AR on the cell surface. These data reveal that beta-ARs function in RPMVECs could be modulated by manipulating beta-ARs traffic from the ER to the Golgi body. We propose the ER-to-Golgi transport as a regulatory site for control of permeability of RPMVECs.

Comparison of inhaled corticosteroid combined with theophylline and double-dose inhaled corticosteroid in moderate to severe asthma.

OBJECTIVE: Recent studies have found that theophylline exerts anti-inflammatory and immunomodulatory effects. This study was performed to compare the efficacy of inhaled corticosteroids (ICS) combined with slow-release theophylline (SRT) with that of double-dose ICS in asthma control, anti-inflammatory activity and safety. METHODOLOGY: In a randomized, open, parallel, control trial, 41 patients with asthma were randomly treated with either beclomethasone dipropionate 500 microg b.i.d. (BDP group) or a combination of BDP 250 microg b.i.d and SRT 0.2 g b.i.d. (SRT/BDP group) for 6 weeks. At the start and at the end of treatment, lung function testing and sputum induction were performed, and plasma cortisol levels were measured. Sputum was analyzed for cell differential counts and the interleukin (IL)-5 level. Patients kept a record of peak expiratory flow (PEF), symptom score, and beta2-agonist use. RESULTS: Significant increases in the morning and the evening PEF and FEV1 were observed (P < 0.05), together with an obvious reduction in symptom score and beta2-agonist use (P < 0.01). Significant decreases in the percentage eosinophils and IL-5 level in induced sputum also occurred (P < 0.05). However, there was no difference between the two groups for all these parameters. There was no significant change in the plasma cortisol level for either group. CONCLUSIONS: Both ICS combined with SRT and double-dose ICS had the same effect on asthma control, improving symptoms and ameliorating lung function. Both therapies had similar anti-airway inflammatory effects and therapeutic safety. Combining SRT with ICS may allow a reduction in ICS dose when treating asthma.