Nasal mucosal fibroblasts produce IL-4 to induce Th2 response.

Th2 polarization is essential for the pathogenesis of allergic rhinitis (AR). Th2 polarization's mechanism requires further understanding. IL-4 is the primary cytokine involved in Th2 response. Fibroblasts play a role in immune regulation. This study aims to elucidate the role of nasal mucosal fibroblast-derived IL-4 in the induction of Th2 responses. Nasal mucosal tissues were obtained from surgically removed samples from patients with nasal polyps, whether with or without AR. Fibroblasts were isolated from the tissues by flow cytometry cell sorting, and analyzed by RNA sequencing (RNAseq). The data from RNAseq showed that nasal fibroblasts expressed genes of GATA3, CD80, CD83, CD86, STAT6, IL2, IL4, IL5, IL6, IL13 and costimulatory factor. The data were verified by RT-qPCR. The level of gene activity was positively correlated with those of AR-related cytokines present in nasal secretions. Nasal fibroblasts release IL-4 upon activation. Nasal fibroblasts had the ability to transform naive CD4+ T cells into Th2 cells, which can be eliminated by inhibiting IL-4 receptor or CD28 in CD4+ T cells. To sum up, nasal mucosal fibroblasts produce IL-4, which can induce Th2 cell development. The data implicate that nasal fibroblasts are involved in the pathogenesis of nasal allergy.

Oligodeoxynucleotides containing CpG motifs (CpG-ODN) restores immune regulatory functions of airway macrophages of patients with asthma.

Allergic asthma is characterized by the polarization of Th2 cells and impaired immune regulation. Macrophages occupy the largest proportion of airway immune cells. This study aims to discover the mechanism that hinders the immune regulatory functions of airway macrophages. In this study, macrophages were isolated from cells in bronchoalveolar lavage fluids (BALF) collected from asthma patients and normal control (NC) subjects. The results indicated that macrophages occupied the largest portion of the cellular components in BALF. The frequency of IL-10+ macrophage was significantly lower in asthma patients than in NC subjects. The expression of IL-10 in macrophages of BALF was associated with the levels of asthma-related parameters. The immune-suppressive functions of BALF M0 cells were defective in asthma patients. The inducibility of IL-10 expression was impaired in BALF macrophages of asthma patients, which could be restored by exposing to CpG. In conclusion, the induction of IL-10 in macrophages of BALF in asthma patients was impaired, and it could be restored by exposure to CpG.

Direct exposure to CpG and specific antigens mitigate airway allergy through modulating dendritic cell properties.

BACKGROUND: CpG oligodeoxynucleotide (CpG-ODN; CpG, in short) has been employed as an adjuvant in allergen specific immunotherapy (AIT) to treat allergic diseases. The underlying mechanism needs to be further explained. The aim of this study is to examine the mechanism by which CpG and dust mite extracts (DME, a specific antigen) alleviate experimental airway allergy. METHODS: DME was used as the specific allergen to establish an airway allergy mouse model. The mice were directly exposed to DME and CpG through nasal instillations (the CpG.DME therapy). The response of DCs and allergic responses in the airways were assessed using immunological approaches. RESULTS: The airway allergy reaction was effectively suppressed by CpG.DME therapy. The administration of CpG or DME alone did not have any significant suppressive effects on the airway allergic response. Direct exposure to CpG.DME induced type 1 DCs (DC1s) and plasmacytoid DCs (pDCs), while CpG alone induced DC1s and DME alone induced DC2s in the airway tissues. Both DC1s and pDCs were required for the induction of type 1 regulatory T cells in the airway tissues by CpG.DME therapy. Depletion of either pDCs or DC1s abolished the induction of Tr1 cells, and abolished the suppressive effects on airway allergic response by the CpG.DME therapy. CONCLUSIONS: Direct exposure to CpG.DME induces DC1s and pDCs in the airway tissues. DC1s in synergy with pDCs induce type 1 regulatory T cells. The CpG.DME therapy is effective in suppressing allergic responses in mice with airway allergy.

Recent developments in the immunopathology of COVID-19.

There has been an important change in the clinical characteristics and immune profile of Coronavirus disease 2019 (COVID-19) patients during the pandemic thanks to the extensive vaccination programs. Here, we highlight recent studies on COVID-19, from the clinical and immunological characteristics to the protective and risk factors for severity and mortality of COVID-19. The efficacy of the COVID-19 vaccines and potential allergic reactions after administration are also discussed. The occurrence of new variants of concerns such as Omicron BA.2, BA.4, and BA.5 and the global administration of COVID-19 vaccines have changed the clinical scenario of COVID-19. Multisystem inflammatory syndrome in children (MIS-C) may cause severe and heterogeneous disease but with a lower mortality rate. Perturbations in immunity of T cells, B cells, and mast cells, as well as autoantibodies and metabolic reprogramming may contribute to the long-term symptoms of COVID-19. There is conflicting evidence about whether atopic diseases, such as allergic asthma and rhinitis, are associated with a lower susceptibility and better outcomes of COVID-19. At the beginning of pandemic, the European Academy of Allergy and Clinical Immunology (EAACI) developed guidelines that provided timely information for the management of allergic diseases and preventive measures to reduce transmission in the allergic clinics. The global distribution of COVID-19 vaccines and emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with reduced pathogenic potential dramatically decreased the morbidity, severity, and mortality of COVID-19. Nevertheless, breakthrough infection remains a challenge for disease control. Hypersensitivity reactions (HSR) to COVID-19 vaccines are low compared to other vaccines, and these were addressed in EAACI statements that provided indications for the management of allergic reactions, including anaphylaxis to COVID-19 vaccines. We have gained a depth knowledge and experience in the over 2 years since the start of the pandemic, and yet a full eradication of SARS-CoV-2 is not on the horizon. Novel strategies are warranted to prevent severe disease in high-risk groups, the development of MIS-C and long COVID-19.

Targeting epithelial cell-derived TWIST1 alleviates allergic asthma.

It is well known that the T Helper (Th)2 bias plays a critical role in allergic asthma. Whereas the Th2 bias is maintained in the local tissues is uncertain. IL-33 is vital for the development of the Th2 polarization. TWIST-1 has an effect on regulating cellular functions. The aberrant activation of RAS sustains certain cellular activities. The aim of this study is to study the role of the interaction between activation of TWIST1 and RAS in inducing and maintaining Th2 polarization in allergic asthma. The epithelial cells of the airways (AEC) were isolated from the broncho-alveolar lavage fluids in patients with asthma. The mediators involved in the over-expression of IL-33 were determined by RNA sequencing. A mouse model was established to test the role of TWIST1 and RAS in developing allergic asthma. We observed a strong expression of TWIST1 in patients with allergic asthma that showed a positive correlation with asthmatic responses. TWIST1 favored the expression of the IL-33 in the AEC. Twist1-deficient AEC-carrying mice did not induce Th2 polarization in the airways. The expression TWIST1 in AECs was positively associated with RAS activation in AECs in patients with allergic asthma. The interaction between RAS and TWIST1 in AECs sustained airway allergic inflammation. Inhibition of TWIST1 or RAS prevented asthma-like inflammation in the mouse airways. In summary, the interaction between TWIST1 and RAS induces and maintains IL-33 expression in AECs to facilitate allergic inflammation in the respiratory tract. Inhibition of TWIST1 or RAS can prevent experimental allergic asthma.

Clinical Evaluation for Sublingual Immunotherapy With Dermatophagoides farinae in Polysensitized Allergic Asthma Patients.

Background: Many studies have demonstrated the efficacy of single-allergen sublingual immunotherapy (SLIT) in polysensitized patients with allergic rhinitis (AR), but less is reported in polysensitized patients with allergic asthma (AS). Method: Data of 133 adult patients with house dust mite (HDM)-induced AS who had been treated for 3 years were collected. These patients were divided into the control group (treated with low to moderate dose of inhaled glucocorticoids and long-acting ß2 agonists, n = 37) and the SLIT group (further treated with Dermatophagoides farinae drops, n = 96). The SLIT group contained three subgroups: the single-allergen group (only sensitized to HDM, n = 35), the 1- to 2-allergen group (HDM combined with one to two other allergens, n = 32), and the 3-or-more-allergen group (HDM combined with three or more other allergens, n = 29). The total asthma symptom score (TASS), total asthma medicine score (TAMS), and asthma control test (ACT) were assessed before treatment and at yearly visits. Forced expiratory volume in 1 s/forced vital capacity (FEV1/FVC) was assessed before treatment and at the end of SLIT. Results: TASS and ACT scores in the control group were significantly higher than that in the single-allergen group and the 1- to 2-allergen group after 1, 2, and 3 years of SLIT and significantly higher than that in the 3-or-more-allergen group after 3-year SLIT (all p < 0.05). TAMS of the control group was significantly higher than that of the other three groups after 0.5, 1, 2, and 3 years of SLIT (all p < 0.05). FEV1/FVC in the control group was significantly higher than baseline after 3 years of immunotherapy (p < 0.05). Conclusion: Patients sensitized to HDM with/without other allergens showed similar efficacy after 3 years of SLIT. However, the initial response of patients with three or more allergens was slower during immunotherapy process.

Assessing Workplace Stress Among Nurses Using Heart Rate Variability Analysis With Wearable ECG Device-A Pilot Study.

This study aims to measure workplace stress of nurses using heart rate variability (HRV) analysis based on data derived from wearable ECG heart rate monitors. The study population consists of 17 nurses at a major public hospital in China. Data was collected from 7 DON nurses (department of neurosurgery; all females; mean age: 31.43 ± 4.50), and 9 ICU nurses (intensive care unit; 8 females and 1 male; mean age: 31.33 ± 5.43). Each participant was asked to wear a wireless ECG heart rate monitor to measure stress level during work, and to complete the Chinese Nurses Stress Response Scale (CNSRS) after work as subjective response criteria. Demographic information, body posture, heart rate, R-R intervals (RRI), low frequency components (LF) and high frequency components (HF) were collected. LF%, LnHF and the squared root of the mean squared differences of successive NN intervals (RMSSD) based on HRV analysis were used to estimate the stress level of nurses. DON nurses reported a higher LF%, lower LnHF and lower RMSSD than ICU nurses. Work shifts were shown to have significant effects on LF%, LnHF and RMSSD respectively, with nurses in long shifts and night shifts reported high stress levels. Higher LF%, lower LnHF and lower RMSSD were found during work shift. Posture analysis revealed negative correlations with LnHF and RMSSD in walking and standing/sitting positions, and a significant negative correlation with LF% in lying-down position. Nurses with higher LF% reported higher CNSRS scores in all subscales, whereas nurses with lower LnHF or RMSSD reported higher CNSRS scores in social phobia and fatigue subscales. The results of this study support the idea that HRV can be used to investigate workplace stress among nurses under real work condition, and can serve as a preventive measure for identifying stress-related illnesses among nurses.

[Rapid determination of fipronil and its metabolite residues in eggs using ultra performance liquid chromatography-tandem mass spectrometry].

A method was developed for the rapid simultaneous determination of fipronil and its metabolite residues in eggs using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The egg samples (2 g) with 2 mL water were extracted with 4 mL acetonitrile. After adding 1 g sodium chloride, the mixture was centrifuged at 9000 r/min for 10 min at 4℃. The suspension was diluted and filtered through a 0.22 µm organic membrane. The analytes were separated on a C18 column (100 mm×2.1 mm, 1.7 µm), and detected under the multiple reaction monitoring (MRM) mode with a negative ESI source. The average recoveries of fipronil and its metabolites varied from 77.4% to 112.1%, and the relative standard deviations were between 4.0% and 13.6% at three spiked levels. The limits of detection (LODs) ranged from 0.10 µg/kg to 0.43 µg/kg. The method is simple, effective, and can be applied to real samples.

[Determination of 250 pesticide residues in vegetables using QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry].

A multiresidue analytical method for the determination of 250 pesticide residues in vegetables was developed by using QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The target compounds were extracted with acetonitrile containing 1% (v/v) acetic acid, purified by a mixed sorbent of MgSO4, primary secondary amine (PSA), graphitized carbon black (GCB) and C18, separated on a Waters ACQUITY™ UPLC BEH C18 column (100 mm x 2. 1 mm, 1.7 µm) and detected by UPLC-MS/MS. Anhydrous magnesium sulfate was used as a dewatering agent. The effects of the amounts of MgSO4, PSA, GCB and C18 added on the recoveries of 250 pesticides were investigated. The results showed that the purification effect was best when 300 mg MgSO4, 200 mg PSA, 10 mg GCB and 100 mg C18 in 2 mL of the extract were added. For the 250 pesticide residues, the limits of detection (LODs) of the method were from 0. 01 to 50. 00 g/kg. The recoveries obtained ranged from 60. 1% to 120% at three spiked levels in Chinese chives with the relative standard deviations between 3. 5% and 19. 5% using matrix matched external standard method. The results showed that the method is able to meet requirements of the multiresidue detection of the 250 pesticides in vegetable. The method has the advantages of rapidity, simplicity, high sensitivity and better purification effect. It is suitable for the rapid determination of the common pesticides in vegetables, and it provides a strong guarantee for the risk assessments of the quality and safety of vegetables.

[Simultaneous determination of 33 primary aromatic amines in polystyrene and polyethylene masterbatches for foods by ultra-performance liquid chromatography-tandem mass spectrometry]].

A comprehensive analytical method based on ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) has been developed for the simultaneous determination of 33 primary aromatic amines (PAAs) in polystyrene (PS) and polyethylene (PE) masterbatches for foods. The PS masterbatches were dissolved with dichloromethane, and methanol was added to precipitate after extraction by ultrasound extraction. Then the extract was purified by passing through a carbon graphite solid phase extraction column. The PE masterbatches were swelled and extracted with dichloromethane by ultrasound. The purified PS solution and PE extract were concentrated, and diluted to 2 mL with methanol-water (1:9, v/v), and filtered through the membranes of 0.22 µm before UPLC-MS/MS analysis. The analytes were separated on a BEH Phenyl column (100 mm x 2.1 mm, 1.7 µm), eluted by gradient with 0.07% (v/v) formic acid in methanol-water (1:9, v/v). The PAAs were detected by UPLC-MS/MS under multiple reaction monitoring (MRM) mode and quantified by the internal standard method. The separation conditions, fragment voltages and collision energies were optimized. The impacts of extraction times, extraction solvents and concentration methods on recoveries were studied. The limits of detection for the 33 primary aromatic amines were 6-10 µg/kg, and the limits of quantitation were 20-30 µg/kg. The mean recoveries of the two different masterbatch products at three spiked levels of 20, 100, 200 µg/kg were 61.3%-119.8%, and the relative standard deviations were 1.4%-14.8%. The experimental results indicated that the method is simple, rapid, sensitive, accurate, and can meet the related requirements for determination.

[Simultaneous determination of 33 primary aromatic amines of free state in fine pigments by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry].

A comprehensive analytical method based on ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) has been developed for the simultaneous determination of 33 primary aromatic amines (PAAs) in fine pigments such as gouache paint, oil painting pigment and acrylic paint. The primary aromatic amines in samples were extracted with acetonitrile. Then the extract was concentrated by centrifugation and nitrogen blow, finally diluted to 2 mL with methanol-water (1:9, v/v) and filtered through 0. 22 im membrane before UPLC-MS/MS analysis. The analytes were separated on a BEH Phenyl column (100 mm x 2. 1 mm, 1. 7 1µm) with 0. 07% (v/v) formic acid in methanol-water as mobile phases in gradient elution. The PAAs were detected by UPLC-MS/MS under multiple reaction monitoring (MRM) mode and quantified by the internal standard method. The separation conditions, fragment voltages and collision energies were optimized. The impacts of extraction times, extraction solvents and concentration methods on recoveries were studied. The limits of detection and limits of quantitation for the 33 primary aromatic amines were 5-50 µg/kg and 15-150 µg/kg respectively. The mean recoveries of three different dye products at three spiked levels were 70. 1% - 115. 8%. The relative standard deviations were 2. 1% - 15%. The expenmental results indicated that the method is simple, rapid, sensitive, accurate and can meet the requirements for the determination.

Simultaneous determination of sex hormones in egg products by ZnCl2 depositing lipid, solid-phase extraction and ultra performance liquid chromatography/electrospray ionization tandem mass spectrometry.

A method was developed for simultaneous determination of residues of 17 sex hormones in egg products. Target compounds were extracted from samples with methanol in an ultrasonic bath, effectively separated from lipids in the extracts by ZnCl(2) depositing filtration and purified using a C(18) solid-phase extraction (SPE) and followed by NH(2) SPE cartridge. The analytes were quantified by liquid chromatography using a BEH C(18) column coupled to an electrospray ionization tandem mass spectrometer (LC-ESI-MS/MS) operating in negative mode for estrogens and in positive multiple reaction monitoring mode for androgens. The parameters of the mass spectrometer and the composition of mobile phase and additives were also optimized to enhance detection sensitivity. Average recoveries of the target compounds varied from 70.0% to 121.0% with relative standard deviations ranging from 2.3% to 11.2% at two fortification levels. The limits of detection (LOD) of the method were from 0.002 µg kg(-1) to 0.23 µg kg(-1) and the limits of quantification (LOQ) were in the range of 0.007-0.76 µg kg(-1).

[Simultaneous determination of seven sex hormones in fish products using ultra performance liquid chromatography-tandem mass spectrometry].

A rapid, specific and highly sensitive method for the determination of seven sex hormones (norgestrel, methyltestosterone, testosterone propionate, medroxyprogesterone acetate, megestrol acetate, chlormadinone acetate, and nandrolone) residues in fish products was developed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with electrospray ionization (ESI) in positive mode. The target compounds were extracted with methanol after the enzyme hydrolysis of the fish products. ZnCl2 was added to the extract solution to remove lipids. Then target compounds were purified by an LC-C18 and an LC-NH2 solid phase extraction cartridges. The target compounds were separated on a Waters ACQUITY UPLC BEH-C18 column (100 mm x 2.1 mm, 1.7 microm) and detected qualitatively and quantitatively in multi reaction monitoring (MRM) mode. For the seven sex hormones, the limits of detection (LOD) of the method were from 0.08 to 0.17 microg/kg and the limits of quantification (LOQ) were in the range of 0.24 -0.58 microg/kg. At the spiked levels of 1 and 4 microg/kg, the average recoveries ranged from 76% to 118% with the relative standard deviations between 5.0% and 11.3% for the seven sex hormones using internal standard method; and the average recoveries ranged from 66% to 94% with the relative standard deviations between 4.5% and 10.7% using matrix matched external standard method. The results showed that both methods are able to meet the multi-residue detection of the seven sex hormone residues in fish products. The degreased large yellow croaker and roast fish fillet real samples from a local market were detected by the developed method, and the seven targets were not found.

[Simultaneous determination of delta-9-tetrahydrocannabinol cannabidiol and cannabinol in edible oil using ultra performance liquid chromatography-tandem mass spectrometry].

A method for the simultaneous determination of delta-9-tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in edible oil was developed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The target compounds were extracted with methanol, purified by an LC-Alumina-N solid phase extraction cartridge, separated and detected by the UPLC-MS/MS. Quantitative analysis was corrected by an isotope internal standard method using delta-9-THC-D3 as internal standard. Average recoveries for the target compounds varied from 68.0% to 101.6% with the relative standard deviations ranging from 7.0% to 20.1% at three spiked levels. The limits of detection (LOD) of the method were from 0.06-0.17 microg/kg and the limits of quantification (LOQ) were in the range of 0.20-0.52 microg/kg. The results showed that the method is able to meet the requirements for the simultaneous determination of THC, CBD and CBN in edible oil.

[Study on simultaneous determination method for banded synthesis hormones residues in egg products].

OBJECTIVE: To develop method of 7 banded synthesis sex hormones residues in egg products determined by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). METHODS: The sample were enzymolied and target compounds were extracted with methanol. ZnCl2 was added to the extract solution to remove lipid and then analytes were purified by LC-C18 and LC-NH2 solid phase extraction cartridge and further determined by UPLC-MS/MS under positive ionization and multiple reaction monitoring (MRM) modes. RESULTS: The limits of detection (LOD) of UPLC-MS/MS method used for testing Chlormadione acetate (CDA), Medroxypogesterone acetate (MPA), Megestrol Acetate (MA), Testosterone propionate (TSP), Norgestrel (NG), Methyltestosterone (MTS) and Nandrolone (NT) in egg products ranged from 0.012 to 0.23 microg/kg, and the limits of quantification (LOQ) were from 0.04 to 0.76 microg/kg. Experiments on spiked samples of egg products showed that at addition level of 2.0 microg/kg, the average recoveries of the sex hormones ranged from 80.2% to 114%, and coefficients of variation from 6.7% to 14.3%; while at addition level of 4.0 microg/kg, the average recoveries ranged from 75% to 119%, and coefficient of variation from 2.9% to 7.3%. CONCLUSION: The method could be able to identify and quantify banded synthesis hormones residues in eggs and egg products. It could be simple and sensitive, suitable for statutory residue testing.