BACKGROUND: Currently, there are no drugs that have been proven to be effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Because of its broad antiviral activity, interferon (IFN) should be evaluated as a potential therapeutic agent for treatment of coronavirus disease 2019 (COVID-19), especially while COVID-19-specific therapies are still under development. METHODS: Confirmed COVID-19 patients hospitalized in the First Affiliated Hospital, School of Medicine, Zhejiang University in Hangzhou, China, from January 19 to February 19, 2020 were enrolled in a retrospective study. The patients were separated into an IFN group and a control group according to whether they received initial IFN-α2b inhalation treatment after admission. Propensity-score matching was used to balance the confounding factors. RESULTS: A total of 104 confirmed COVID-19 patients, 68 in the IFN group and 36 in the control group, were enrolled. Less hypertension (27.9% vs. 55.6%, P=0.006), dyspnea (8.8% vs. 25.0%, P=0.025), or diarrhea (4.4% vs. 19.4%, P=0.030) was observed in the IFN group. Lower levels of albumin and C-reactive protein and higher level of sodium were observed in the IFN group. Glucocorticoid dosage was lower in the IFN group (median, 40 vs. 80 mg/d, P=0.025). Compared to the control group, fewer patients in the IFN group were ventilated (13.2% vs. 33.3%, P=0.015) and admitted to intensive care unit (ICU) (16.2% vs. 44.4%, P=0.002). There were also fewer critical patients in the IFN group (7.4% vs. 25.0%, P=0.017) upon admission. Although complications during admission process were comparable between groups, the discharge rate (85.3% vs. 66.7%, P=0.027) was higher and the hospitalization time (16 vs. 21 d, P=0.015) was shorter in the IFN group. When other confounding factors were not considered, virus shedding time (10 vs. 13 d, P=0.014) was also shorter in the IFN group. However, when the influence of other factors was eliminated using propensity score matching, virus shedding time was not significantly shorter than that of the control group (12 vs. 15 d, P=0.206). CONCLUSIONS IFN-α2b spray inhalation did not shorten virus shedding time of SARS-CoV-2 in hospitalized patients.
Albumins/analysis* ; Antiviral Agents/administration & dosage* ; Betacoronavirus ; C-Reactive Protein/analysis* ; COVID-19 ; Case-Control Studies ; China ; Coronavirus Infections/drug therapy* ; Glucocorticoids/pharmacology* ; Hospitalization ; Humans ; Interferon alpha-2/administration & dosage* ; Nasal Sprays ; Pandemics ; Pneumonia, Viral/drug therapy* ; Propensity Score ; Retrospective Studies ; SARS-CoV-2 ; Sodium/blood* ; Virus Shedding/drug effects* ; COVID-19 Drug Treatment

OBJECTIVETo observe serum osteoprotegerin (OPG) level in children with nephrotic syndrome (NS) and changes in serum OPG level after glucocorticoid therapy, with the aim of studying the role of OPG in the bone metabolism of children with NS.

METHODSForty-four children with idiopathic NS were randomly selected as the study group, including 24 newly diagnosed, untreated patients and 20 who had relapsed during the process of glucocorticoid reduction (cumulative dose of glucocorticoid 28327±5879 mg/m2). Twenty-three age- and sex-matched healthy children served as the control group. Serum osteoprotegerin (OPG) level was measured using ELISA. Serum N-terminal midfragment of osteocalcin (N-MID osteocalcin) was determined using electrochemical luminescence immunoassays (ECLIA).

RESULTSSerum levels of OPG (211±55 ng/L) and N-MID osteocalcin (46±14 ng/mL) in the untreated NS group were reduced compared with 470±57 ng/L (OPG) and 73±9 ng/ml (N-MID osteocalcin) in the control group (P<0.05). Serum levels of OPG (176±42 ng/L) and N-MID osteocalcin (29±10 ng/mL) in the NS relapsed group were lower than in the untreated NS and control groups (P<0.05).

CONCLUSIONSBone metabolism disorders are found in children with NS. High-doses of glucocorticoid therapy can aggravate these disorders. Serum OPG levels in children with NS may be affected by both the renal disease itself and steroid therapy, suggesting that OPG is expected to become a new biochemical indicator for predicting changes to the bone metabolism of children with NS.

Child ; Glucocorticoids ; pharmacology ; Humans ; Nephrotic Syndrome ; blood ; drug therapy ; Osteocalcin ; blood ; Osteoprotegerin ; blood

INTRODUCTIONGlucocorticoids cause osteoporosis by decreasing bone formation and increasing bone resorption activity. Glucocorticoid action in bones depends on the activity of 11-beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1) enzyme, which plays an important role in regulating corticosteroids. 11β-HSD1 is expressed by human and rat osteoblasts. We aimed to investigate the relationship between 11β-HSD1 dehydrogenase activity and bone histomorphometric changes in glucocorticoid-induced osteoporotic bone in rats.

METHODSA total of 30 male Sprague-Dawley rats (aged three months, weighing 200-250 g) were divided into three groups of ten each. Group 1 rats were the baseline control, which were sacrificed untreated at the beginning of the study. Group 2 rats underwent sham operation and were administered with vehicle olive oil intramuscularly at 0.05 ml/kg. Group 3 rats were adrenalectomised and administered with an intramuscular injection of dexamethasone 120 μg/kg body weight/day. The treatment was started two weeks after the operation, for a duration of two months. Plasma osteocalcin, plasma pyrodinoline, plasma corticosterone and 11β-HSD1 were measured, and bone histomorphometry analysis was performed.

RESULTSDexamethasone treatment caused an increase in plasma corticosterone level, together with a significant reduction in 11β-HSD1 dehydrogenase activity of the bone, along with a higher plasma level of the bone resorption marker, pyridinoline. Dexamethasone treatment also caused a reduction in trabecular volume, number and thickness, and an increase in trabecular separation.

CONCLUSIONLong-term glucocorticoid treatment reduces the 11β-HSD1 dehydrogenase activity in the bone, which can otherwise lead to bone loss due to the increased level of active glucocorticoids.

11-beta-Hydroxysteroid Dehydrogenase Type 1 ; metabolism ; Adrenal Cortex Hormones ; metabolism ; Amino Acids ; pharmacology ; Animals ; Body Weight ; Bone and Bones ; metabolism ; Corticosterone ; blood ; Dexamethasone ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; methods ; Gene Expression Regulation, Enzymologic ; Glucocorticoids ; metabolism ; Humans ; Male ; Osteoporosis ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish rat models of Steroid-avascular necrosis of femoral head, observe the effects of activating blood circulation of chinese herbal medicine on genetic expression of transforming growth factor-beta1 (TGF-beta1). To interpret the mechanism of the effect on Steroid-avascular necrosis of femoral head by activating blood circulation,and offer a effective method to clinical.

METHODSCleaner-40 SD rats,half males and half females, weight (200 +/- 20) g, were randomly divided into 2 groups: 4 rats were in common group and 36 rats were in medel group. The rats in medel group were administered with 24.5 mg/kg hydroxyprednisone twice a week peritoneal injection for 6 weeks induced to femur head necrosis. The rats in common group were through gluteus injection as control. There were 4 rats were killed in each group after 6 weeks, to be assure that the model were succed. All surplus rats were divided into treatment group and control group:the treatment group were administrated with activating blood circulation of Chinese herbal medicine 12.3 ml/kg per day,the control group were administrated with sodium chloride 12.3 ml/kg per day. Then, after 6 and 8 weeks, killed the animal and detected all indexes.

RESULTS(1) The expression of TGF-beta by immunohistochemistry and image analysis: the expression of femoral head TGF-beta increased significantly in treatment group than in control group and two group had significant differences. (2) Serum levels of TGF-beta1: in the treatment group serum expression of TGF-beta1 increased,compared with the control group had significant difference (P < 0.01). (3) The femoral head local TGF-beta1 mRNA transcription: treatment group in the first six weeks, expressed that the increase in the first eight weeks, expressed also reduced,and the control group in the first six weeks, expressed a decrease in the first eight weeks,was not detected to TGF-beta1 mRNA expression of difference between the two groups was significant (P < 0.01). TGF-beta1 mRNA in serum and the femoral head with local expression of TGF-beta1, in consistency.

CONCLUSIONRat abdominal cavity prednisolone acetate injection plus intermittent standing avascular necrosis modeling stability, good repeatability. Taohongsiwu through the promotion of hormone-ischemic necrosis of the femoral head rat model of TGF-beta1 mRNA transcription, and promote expression of TGF-beta1.

Animals ; Blood Circulation ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; Glucocorticoids ; pharmacology ; Male ; Medicine, Chinese Traditional ; Rats ; Transforming Growth Factor beta1 ; blood ; genetics ; physiology

OBJECTIVETo identify the relationship between the expression of alpha and beta-isoforms of glucocorticoid receptors (GR) in peripheral blood mononuclear cells (PBMC) and glucocorticoid resistance in children with idiopathic thrombocytopenia purpura (ITP).

METHODSReal-time PCR was used to detect the expression of GR alpha and GR beta mRNA in PBMC from 30 children with ITP (glucocorticoid-sensitive, n=18; glucocorticoid-resistant, n=12) and 10 healthy children (control group). Enzyme immunoassay was used to measure plasma levels of total glucocorticoids.

RESULTSThere were no significant differences in PBMC GR alpha mRNA levels among the glucocorticoid sensitive, the glucocorticoid-resistant and the control groups. Compared with the glucocorticoid-sensitive and the control groups, the expression of GR beta mRNA in the glucocorticoid-resistant group was significantly up-regulated (p<0.01). Plasma total glucocorticoids level in the glucocorticoid-resistant group was found to be much higher than that in the glucocorticoid-sensitive and the control groups (p<0.01).

CONCLUSIONSThe up-regulated expression of GR beta mRNA may associated with glucocorticoid resistance in children with ITP.

Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; pharmacology ; Humans ; Male ; Purpura, Thrombocytopenic, Idiopathic ; blood ; drug therapy ; RNA, Messenger ; analysis ; Receptors, Glucocorticoid ; blood ; genetics

OBJECTIVE: The purpose of this study is to evaluate the effect of dexamethasone on the damaged blood-ocular barrier caused by triolein emulsion, using contrast-enhanced MR imaging. MATERIALS AND METHODS: An emulsion of 0.1-mL triolein in 20 mL of saline was infused into the carotid arteries of 32 cats, 12 cats were placed in the treatment group and 18 cats were placed in the Control group. Thirty minutes after the infusion of triolein emulsion, a set of orbital pre- and post-contrast T1-weighted MR images (T1WIs) were obtained. Infusion of 10 mg/kg dexamethasone into the ipsilateral carotid artery of each of the cats in the treatment group cats and 20 mL saline in each of the cats in the control group was given. A second set of pre- and post-contrast orbital T1WIs were obtained three hours following triolein emulsion infusion. Qualitative analysis was performed for the the anterior chamber (AC), the posterior chamber (PC), and in the vitreous humor of the ipsilateral and contralateral eyes. The signal intensity ratios of the ipsilateral eye over the contralateral eye were quantitatively evaluated in the three ocular chambers on the first and second set of T1WIs, and were then statistically compared. RESULTS: Qualitatively, the AC, the PC or the vitreous did not show immediate contrast enhancement on the first and the second set of post-contrast T1WIs. However, the AC and the PC showed delayed contrast enhancement for both groups of cats on the second pre-contrast T1WIs. No enhancement or minimally delayed enhancement was seen for the vitreous humor. Quantitatively, the signal intensity ratios in the PC of the treatment group of cats were statistically lower than the ratios of the control group of cats for the second set of T1WIs (p = 0.037). The AC and vitreous showed no statistically significant difference between the feline treatment group and control group (p > 0.05). CONCLUSION: Contrast-enhanced MR images revealed increased vascular permeability in the PC of the eye after infusion of triolein emulsion. Dexamethasone seems to decrease the breakdown of the blood-aqueous barrier in the PC.
Animals ; Blood-Aqueous Barrier/*drug effects ; Blood-Retinal Barrier/*drug effects ; Capillary Permeability/drug effects ; Cats ; Contrast Media ; Dexamethasone/*pharmacology ; Emulsions ; Glucocorticoids/*pharmacology ; Image Enhancement ; Magnetic Resonance Imaging/*methods ; Triolein/*adverse effects

OBJECTIVETo determine the changes of CD4+CD25+ regulatory T cells and the levels of IL-10 and TGF-beta1 in the mouse model of asthma and the effects of glucocorticoid inhalation on CD4+CD25+ regulatory T cells and IL-10 and TGF-beta1 levels.

METHODSThirty BALB/c mice were randomly divided into three groups: asthma model, glucocorticoid treatment and control. Asthma was induced by OVA administration in the asthma model and the glucocorticoid treatment groups. The glucocorticoid treatment group received budesonide inhalation (0.8 mg/L) before the challenge of asthma. After 24 hrs of the last challenge, the lung tissues of middle lobe of the right lung were obtained for the observation of lung pathological changes. Peripheral anticoagulated blood and lung lymph cells suspension were collected. The percentage of CD4+CD25+ regulatory T cells in CD4+ T cells was detected by flow cytometry, and the levels of IL-10 and TGF-beta1 in plasma were measured using ELISA.

RESULTSThe percentage of CD4+CD25+ regulatory T cells in peripheral blood and lung lymph cells suspension in the asthma model group was significantly lower than the control group ( P<0.01). The glucocorticoid-treated asthma group showed an increased percentage of CD4+CD25+ regulatory T cells compared with the asthma model group (P<0.01) and a similar percentage of CD4+CD25+ regulatory T cells in peripheral blood and lung lymph cells suspension to the control group. Compared with the control group, plasma levels of IL-10 and TGF-beta1 decreased significantly in the asthma model group (P<0.01). After glucocorticoid treatment plasma IL-10 level increased significantly (P<0.01) and was similar to that in the control group, while plasma TGF-beta1 level remained lower than that in the control group.

CONCLUSIONSThe percentage of CD4+CD25+ regulatory T cells and the levels of IL-10 and TGF-beta1 decreased in asthmatic mice which might contribute to the pathogenesis of asthma. Glucocorticoid can increase the percentage of CD4+CD25+ regulatory T cells and the levels of IL-10 and thus provides protective effects against asthma. The changes of the percentage of CD4+CD25+ regulatory T cells in peripheral blood were consistent with those in the lung, suggesting that monitoring the changes of CD4+CD25+ regulatory T cells in peripheral blood may be useful to understand the changes of CD4+CD25+ regulatory T cells in the lung.

Animals ; Asthma ; drug therapy ; immunology ; Disease Models, Animal ; Female ; Glucocorticoids ; pharmacology ; Interleukin-10 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Regulatory ; drug effects ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo study the variation of TFAR-19 protein expression and Annexin-V apoptosis protein proportion in the mice thymus cell apoptosis procedure induced by water platform environment or electric emotional stress and the regulatory effect of Modified Xiaoyao Pill (MXP).

METHODSThe mouse emotional stress model was established by water platform equipment or electrical stimulation. The serum glucocorticoid was detected by radioimmunoassay, the TFAR-19 protein was detected by flow cytometry analysis, and Annexin-V apoptosis protein proportion was calculated by immunofluorescence technique.

RESULTSIn the groups of mouse stressed by water platform environment, the level of serum glucocorticoid, the TFAR-19 protein expression and the Annexin-V apoptosis protein proportion increased in the thymus cell along with the stress time prolonging (P<0.05 or P <0.01). The serum glucocorticoid level in mice treated with MXP was lower than that in the untreated group (P <0.05). In the groups of emotional stressed mouse established by electrical stimulation, the above-mentioned variations also revealed. All these variations could be alleviated with MXP (P<0.05).

CONCLUSIONThe water platform environment stress is a chronic continuous stress and electrical stimulation is an acute smooth stress, both of them could damage thymus function through neuro-endocrineo-immune network, but different in duration for causing severe injury. Chinese medicine MXP can alleviate the damage of thymus induced by either of them to certain degree.

Animals ; Annexin A5 ; biosynthesis ; Apoptosis Regulatory Proteins ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glucocorticoids ; blood ; Male ; Mice ; Neoplasm Proteins ; biosynthesis ; Stress, Psychological ; blood ; physiopathology ; Thymus Gland ; cytology ; drug effects ; metabolism

OBJECTIVEMany studies have shown that glucocorticoids play a crucial role in the development of obesity and insulin resistance. This study investigated the therapeutic effects of long-term inhibition of glucocorticoid activity on obesity and insulin resistance.

METHODSFour-week-old male Sprague-Dawley (SD) rats were randomly fed with a high-fat diet (fat content accounting for 20% of total calorie) (control group, n=8) or with a high-fat diet along with glycyrrhetic acid (GE, 800 mg/L), an inhibitor of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) for 24 weeks (GE-treated group, n=9). The body weights and the amount of food intake were monitored weekly and daily, respectively. After 24 weeks of GE treatment, oral glucose tolerance tests were performed. Blood glucose was measured by glucose oxidase method. The levels of plasma glucocorticoids, insulin and leptin were measured with radioimmunoassay. The levels of serum cholesterol and triglyceride were determined with an automatic measuring analyzer.

RESULTSThe food intake amount decreased significantly in the GE-treated group from 6 weeks and body weight gain was markedly less from 8 weeks after GE administration compared with the control group. After 24 weeks of treatment, the plasma levels of leptin and insulin in GE-treated rats were significantly reduced compared with the control group. The serum levels of cholesterol and triglyceride decreased markedly compared with the control group and the levels of blood glucose were significantly lower 15, 30, 60 and 120 minutes after oral glucose load in the GE-treated group compared with the control group.

CONCLUSIONSLong-term GE treatment may contribute to resisting diet-induced obesity and insulin resistance.

11-beta-Hydroxysteroid Dehydrogenases ; antagonists & inhibitors ; Animals ; Body Weight ; drug effects ; Dietary Fats ; administration & dosage ; Enzyme Inhibitors ; pharmacology ; Glucocorticoids ; physiology ; Glucose Tolerance Test ; Glycyrrhetinic Acid ; pharmacology ; therapeutic use ; Insulin ; blood ; Insulin Resistance ; Leptin ; blood ; Male ; Obesity ; drug therapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVEEosinophilic airway inflammation is one of the basic characteristics of allergic asthma. Toll-like receptor is one of the most important innate immunity pattern recognition receptors. Glucocorticoids (GCS) are still the most effective treatment for asthma. However, few reports of studies on regulatory mechanism of GCS on the innate immunity system are available. The mechanism of effects of GCS on TLR4 is unclear. The present study aimed at understanding the effect of dexamethasone (DXM) on change of TLR4 and mechanism of regulatory effect of TLR4 on eosinophil (EOS) apoptosis.

METHODSTwenty-seven Sprague-Dawley (SD) rats (age 28 to 42 days, body weight 120 to 180 gram) were randomly divided into the control group, asthma group and DXM group with 9 in each. Asthma model rats were sensitized with the mixture of ovalbumin (OVA, 1 mg) and Al (OH)(3), 100 mg on day 1 and day 8, repeatedly exposed to aerosolized OVA after day 15, once a day for three days and continued for 30 minutes at every time. During the sensitization stage, 100 microg/ml DXM were prepared with DXM group for every other day, and the same doses DXM were prepared for every day on the stage of challenge. The histopathological changes of lung tissues were observed with light microscope (LM). EOS and other inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted; the concentrations of OVA-sIgE in serum were measured by using "sandwich" ELISA; The expressions of TLR4 mRNA were determined by in situ hybridization, the apoptosis of EOS was detected by TUNEL.

RESULTS(1) LM showed many inflammatory cells infiltration around the bronchi and blood vessels, bronchus mucus increased, airway epithelium damage and desquamation, and airway mucous plugs in asthma group, whereas DXM group showed significantly milder changes. (2) Inflammationary cells count in BALF of asthma group was significantly higher as compared to control group (P < 0.01); compared with asthma group, the total cell count, EOS absolute count and EOS% were all significantly decreased in DXM group [(2.14 +/- 0.10) x 10(9)/L, (4.78 +/- 1.23) x 10(7)/L, (2.17 +/- 0.25)%]. (3) Levels of OVA-sIgE in serum of asthma group [(83.40 +/- 6.80) microg/ml] were significantly higher than those of the control group [(14.38 +/- 4.25) microg/ml] (P < 0.01), while those of DXM group [(45.02 +/- 7.47) microg/ml] were significantly lower than asthma group (P < 0.0 1). (4) There were no significant differences in TLR4 mRNA detected by in situ hybridization between control group (24.71 +/- 0.85) and asthma group (25.81 +/- 3.56) (P > 0.05); but it significantly increased in DXM group (29.86 +/- 3.92) as compared to asthma group. (5) The percentages of apoptotic EOS in asthma group [(7.39 +/- 1.93)%] were significantly lower than those in control group [(9.06 +/- 1.52)%] (P < 0.01); and significantly higher in DXM group [(13.33 +/- 1.09)%] than in asthma group (P < 0.01). There were significantly positive correlations between TLR4 mRNA and the percentage of apoptotic EOS (r = 0.612, P < 0.01).

CONCLUSIONDXM can decrease OVA-sIgE level, induce EOS apoptosis, which may correlate with the activation of TLR4 signal transduction.

Animals ; Apoptosis ; Asthma ; chemically induced ; immunology ; Bronchoalveolar Lavage Fluid ; cytology ; Dexamethasone ; pharmacology ; Eosinophils ; immunology ; Glucocorticoids ; pharmacology ; Immunoglobulin E ; blood ; Lung ; pathology ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; immunology ; metabolism