Pyroptosis-Related Genes as Diagnostic Markers in Chronic Obstructive Pulmonary Disease and Its Correlation with Immune Infiltration.

Background: Chronic obstructive pulmonary disease (COPD) stands as a predominant cause of global morbidity and mortality. This study aims to elucidate the relationship between pyroptosis-related genes (PRGs) and COPD diagnosis in the context of immune infiltration, ultimately proposing a PRG-based diagnostic model for predicting COPD outcomes. Methods: Clinical data and PRGs of COPD patients were sourced from the GEO database. The "ConsensusClusterPlus" package was employed to generate molecular subtypes derived from PRGs that were identified through differential expression analysis and LASSO Cox analysis. A diagnostic signature including eight genes (CASP4, CASP5, ELANE, GPX4, NLRP1, GSDME, NOD1and IL18) was also constructed. Immune cell infiltration calculated by the ESTIMATE score, Stroma scores and Immune scores were also compared on the basis of pyroptosis-related molecular subtypes and the risk signature. We finally used qRT - PCR to detect the expression levels of eight genes in COPD patient and normal. Results: The diagnostic model, anchored on eight PRGs, underwent validation with an independent experimental cohort. The area under the receiver operating characteristic (ROC) curves (AUC) for the diagnostic model showcased values of 0.809, 0.765, and 0.956 for the GSE76925, GSE8545, and GSE5058 datasets, respectively. Distinct expression patterns and clinical attributes of PRGs were observed between the comparative groups, with functional analysis underscoring a disparity in immune-related functions between them. Conclusion: In this study, we developed a potential as diagnostic biomarkers for COPD and have a significant role in modulating the immune response. Such insights pave the way for novel diagnostic and therapeutic strategies for COPD.

Growth performance, reproductive traits and offspring survivability of genetically modified rams overexpressing toll-like receptor 4.

Genetic modification provides a means to enhancing disease resistance in animals. Toll-like receptor 4 (TLR4), a member of the TLR family, is critical for the recognition of lipopolysaccharide (LPS)/endotoxin from Gram-negative bacteria by host immune cells, which initiates cell activation and subsequently triggers a proinflammatory response to the invading pathogens. In this study, the first generation of genetically modified (GM) sheep overexpressing TLR4 was produced by microinjection for better disease resistance. Compared with wild-type (WT) rams, the GM rams have similar growth performance, basic semen quality and spermatozoon ultrastructure. The offspring birth rates after cervical artificial insemination were also similar between GM (90.32%) and WT (92.38%) rams. Overall, the presence and expression of the TLR4 transgene in the genome did not appear to interfere with normal semen production, reproductive traits and the ability of transgene transmission to offspring. The expression levels of TLR4, tumor necrosis factor and interferon gamma genes in monocyte/macrophages from GM sheep were significantly higher than that from WT sheep at early stages after LPS stimulation. The GM offspring born from the founder transgenic ram inseminated ewes had similar survival rate with WT offspring (88.89% vs 84.86%) at weaning. The TLR4 transgene showed no deleterious effects on growth performance, reproductive traits and offspring survivability of GM rams. Therefore, the GM sheep overexpressing TLR4 provide a powerful experimental model for analyzing function of TLR4 in vivo during infection and inflammation.

Comparison and evaluation of several Dermatophagoides pteronyssinus allergen extracts for skin prick test.

OBJECTIVE: To evaluate the significance of several Dermatophagoides pteronyssinus allergen extracts for skin prick test (SPT) in patients allergic to Dermatophagoides pteronyssinus. METHODS: Two hundred and nineteen patients enrolled in Peking Union Medical College Hospital underwent SPT and serum specific IgE assay to detect the Dermatophagoides pteronyssinus allergen. Three kinds of house dust mite allergen extracts were used for SPT, including the Dermatophagoides pteronyssinus extract prepared by our laboratory (group A), standardized Dermatophagoides pteronyssinus extract (group B), and mixed extracts of Dermatophagoides pteronyssinus and Dermatophagoides farinae (group C). Human serum specific IgE result was regarded as the reference standard for diagnosis of Dermatophagoides pteronyssinus allergy. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of SPT with the extracts of three groups. RESULTS: SPT results showed that the median wheal diameter of group A, group B, and group C was 0.43, 0.35, and 0.28 cm, respectively, with significant difference among three groups (P<0.05). The difference was significant between group A and B (P<0.01) as well as group A and C (P<0.01), but not between group B and C (P>0.05). There was no local urticaria or systemic allergic reactions following the procedure of SPT. Local reaction was observed in 5 patients and delayed reaction was in 2 patients of group A. As for group B and C, local reaction occurred in 3 cases and delayed reaction in 2 cases in each group. The area under ROC curve of SPT with extract in group A, group B, and group C was 0.765, 0.801, and 0.782, respectively. Based on the detection results of serum specific IgE, the sensitivity of SPT in diagnosis of Dermatophagoides pteronyssinus allergy with extract of group A, group B, and group C was 92.4%, 87.0%, and 81.5%, and the specificity was 60.6%, 73.2%, and 74.8%, respectively. CONCLUSION: The Dermatophagoides pteronyssinus extract for SPT prepared by our laboratory offers good sensitivity and specificity comparable to commercially available allergen extracts, and it may be an appropriate candidate for clinical screening and diagnosis of Dermatophagoides pteronyssinus allergy.

[Values of intradermal skin test and serum sIgE detection in diagnosing Humulus scandens pollinosis].

OBJECTIVE: To evaluate the values of intradermal skin test (IDT) and serum specific IgE detection in diagnosing Humulus scandens sensitivity in Chinese patients with autumnal hay fever. METHODS: 1150 patients, 504 males and 646 females, aged 5 approximately 75, were evaluated as with autumnal rhinitis and asthma by experienced physicians. Then the patients underwent IDT by using 20 kinds of aeroallergen extracts, at the concentration of 1:1000 (w/v) for Artemisia spp. and ragweed or at the concentration of 1:100 (w/v) for Humulus scandens and others. The Humulus sIgE level was performed in all patients. A diagnosing criteria was established according to typical history, symptoms, and any one of the following findings: (1) wheal > or = 5 mm in diameter by IDT and sIgE to Humulus of the concentration > or = 0.35 kUa/L at same time; (2) wheal > or = 10 mm in diameter by IDT alone; and (3) sIgE to Humulus of the concentration > or = 0.70 kUa/L alone. RESULTS: When using the above diagnosing criteria as the reference standard, IDT had better sensitivity (97.2%), positive predictive value (PV+) (77.9%), negative predictive value (PV-) (90.6%) and efficiency (80.4% than using sIgE > or = 0.35 kUa/L alone as the reference criteria of IDT, however, had a lower specificity (49.8%); and sIgE detection had better sensitivity (89.0%), specificity (97.5%), PV+ (98.8%), PV- (88.0%) and efficiency (92.0%) than using wheal > or = 5 mm in diameter alone as the reference criteria of sIgE detection. The false positive rate of IDT decreased from 59.9% to 17.4% when using wheal diameter > or = 10 mm as the positive criteria. In the 288 patients with a negative sIgE result and a positive IDT result 84 cases had stronger positive IDT result (with the wheal diameter > or = 10 mm). CONCLUSION: IDT is correlated well with sIgE detection in diagnosing Humulus scandens pollinosis, but the false positive rate of IDT is higher than that of sIgE test. The false positive rate of IDT can be decreased by increasing the positive criteria to higher grading reaction. The immunotherapy according to the lower grading positive result of IDT alone or sIgE alone should be avoided.

[Value of intradermal skin test and serum sIgE detection in diagnosing artemisia pollinosis].

OBJECTIVE: To evaluate the value of intradermal skin test (IDT) and serum sIgE detection in diagnosing Artemisia sensitivity in Chinese patients with autumnal hay fever. METHODS: 1150 patients with autumnal rhinitis or asthma, 504 males and 646 females, aged 5 approximately 77, were evaluated by experienced physicians, then underwent IDT by using 20 kinds of aeroallergen extracts. The concentrations of Artemisia and Ragweed extracts employed in skin test were 1:1000 (W/V) and the concentrations of other aeroallergens were all 1:100 (W/V). Then all patients underwent detection of Artemisia sIgE. Diagnostic standards were established based on the results of IDT and sIgE results respectively. A reference standard was established according to the typical history, symptoms, and an wheal with a diameter >or= 5mm and a sIgE level >or= 0.35 kU(A)/L, an wheal with the diameter >or= 10 mm alone; or a sIgE level >or= 0.70 kUa/L alone. RESULTS: When using the reference standard as criteria, using IDT had better sensitivity (96.2%), specificity (74.2%), positive predictive value (+PV, 93.5%), negative predictive value (-PV, 85.7%), and efficiency (91.6%) than using sIgE >or= 0.35 kUa/L alone as the criteria of IDT; sIgE detection had better sensitivity (97.6%), specificity (94.9%), +PV (98.7%), -PV (91.1%), and efficiency (97.0%) than using wheal diameter >or= 5 mm alone as the criteria of sIgE detection. The false positive rate of IDT and sIgE detection decreased from 35% and 22.7% to 25.6% and 5.1% respectively when using the wheal diameter >or= 10 mm or sIgE >or= 0.70 kUa/L as a positive criteria. CONCLUSION: IDT and sIgE detection are correlated with each other well in diagnosing Artemisia pollinosis, both of them have the possibility of being false positive, but IDT has higher false positive rate than sIgE detection. The false positive rates of IDT and sIgE detection can be decreased by increasing the positive criteria to higher grading reaction.

[Natural course from rhinitis to asthma in the patients with autumnal pollinosis: a clinical study of 1096 patients].

OBJECTIVE: To investigate the natural course from rhinitis to asthma in the patients with autumnal pollinosis. METHODS: 1096 patients with autumnal pollinosis, 488 males and 632 females, aged 38 +/- 14 (5 - 77), 511 with pure allergic rhinitis and 585 with allergic rhinitis complicated with asthma, underwent skin test of inhalant allergens, serum IgE specific to autumnal pollens, and questionnaire survey. RESULTS: The age range of rhinitis induced by autumnal pollens was 2 - 75. The peak onset age of rhinitis was 15 - 44, and the peak onset age of asthma was 25 - 54. 33% of the rhinitis patients complicated with asthma (194/585) had their first attack of rhinitis and the first attack of asthma in the same year, 66% of them (386/585) had the first attack of asthma later than the first attack of rhinitis, and only 0.8% of them (5/585) had their first attack of asthma earlier than that of rhinitis. 37% of the patients with autumnal pollen allergic rhinitis (410/1096) developed asthma within 5 years, 47% (511/1096), within 9 years, 5% (58/1096) within 10 approximately 19 years, and 1.5% (16/1096) within 20 - 40 years. CONCLUSION: Almost half of the patients with autumnal pollen allergic rhinitis develop to seasonal allergic asthma within 9 years.

[The clinical study of the relationship between allergic rhinitis and allergic asthma in the patients with autumnal pollinosis].

OBJECTIVE: To investigate the relationship between allergic rhinitis and allergic asthma in the patients with autumnal pollinosis. METHODS: 1120 patients with autumnal pollinosis, aged 5 - 75, excluding those with typical symptoms of seasonal rhinitis or asthma but with positive skin test and serum IgE specific to dustmite and fungi, underwent standardized clinical questionnaire survey, including the onset age, onset time, and symptoms as well as the severity of asthma, skin tests, and examination of serum IgE specific to autumnal pollens. RESULTS: The average onset age of the allergic rhinitis patients induced by autumnal pollens was 27.9 years, significantly younger than that of the allergic asthma patients (32.6 years, P < 0.001). Out of the 1120 patients 1096 (97.9%) had allergic rhinitis, 602 (53.8%) had asthma, 507 (45.3%) had allergic rhinitis only, and 10 (0.9%) had allergic asthma only. Among the 1096 patients with allergic rhinitis 585 (53.4%) suffered from seasonal asthma. Among the 602 patients with asthma 585 (97.2%) suffered from seasonal rhinitis, and 183 of the 602 patients (30.8%) needed emergency treatment. CONCLUSION: Autumnal pollens are very important causes which induce asthma during autumnal season in northern China.

Effects of SDS, PBS solutions upon fluorescence values of Dermatophagoides pterronyssinus radioallergosorbent test inhibition.

OBJECTIVE: To explore the effects of SDS, PBS re-dissolvent solutions on fluorescence values of radioallergosorbent test (RAST) inhibition. METHODS: Dermatophagoides pterronyssinus allergen immunoCAP and UniCAP 100 System were used. The Sera Pool consisted of 20 Dermatophagoides pterronyssinus allergic patients sera, their specific IgE fluorescence values were between 12505 and 24776. RESULTS: Fluorescence value percentages decreased: 62.9%, 54.1%, 43.5%, 6.7%, 3.7%, 2.6%, 2.2%, and 1.4% respectively, when SDS concentrations were at 2%, 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.025%, and 0.01%. Fluorescence values decreased more than 5% with SDS concentrations equal to 0.25% or higher. PBS in 0.1 and 0.01 mol/L concentrations decreased fluorescence values 2.9% and 0.9% respectively. CONCLUSIONS: SDS is a commonly used surfactants in allergen extract and re-dissolvent prepared allergen precipitation for RAST inhibition. Thus effects of surfactants (e.g. SDS) upon the RAST inhibition tests must be considered when they were used as re-dissolvent agents to improve protein resolution in RAST inhibition.