Visualizing Low-Abundance Proteins and Post-Translational Modifications in Living Drosophila Embryos via Fluorescent Antibody Injection.

Visualization of proteins in living cells using GFP (Green Fluorescent Protein) and other fluorescent tags has greatly improved understanding of protein localization, dynamics, and function. Compared to immunofluorescence, live imaging more accurately reflects protein localization without potential artifacts arising from tissue fixation. Importantly, live imaging enables quantitative and temporal characterization of protein levels and localization, crucial for understanding dynamic biological processes such as cell movement or division. However, a major limitation of fluorescent tagging approaches is the need for sufficiently high protein expression levels to achieve successful visualization. Consequently, many endogenously tagged fluorescent proteins with relatively low expression levels cannot be detected. On the other hand, ectopic expression using viral promoters can sometimes lead to protein mislocalization or functional alterations in physiological contexts. To address these limitations, an approach is presented that utilizes highly sensitive antibody-mediated protein detection in living embryos, essentially performing immunofluorescence without the need for tissue fixation. As proof of principle, endogenously GFP-tagged Notch receptor that is barely detectable in living embryos can be successfully visualized after antibody injection. Furthermore, this approach was adapted to visualize post-translational modifications (PTMs) in living embryos, allowing the detection of temporal changes in tyrosine phosphorylation patterns during early embryogenesis and revealing a novel subpopulation of phosphotyrosine (p-Tyr) underneath apical membranes. This approach can be modified to accommodate other protein-specific, tag-specific, or PTM-specific antibodies and should be compatible with other injection-amenable model organisms or cell lines. This protocol opens new possibilities for live imaging of low-abundance proteins or PTMs that were previously challenging to detect using traditional fluorescent tagging methods.

Clinical Efficacy and Safety of Tumor Cytoreductive Surgery plus Hyperthermic Intraperitoneal Chemotherapy for Ovarian Cancer.

Objective: To assess the clinical efficacy and safety of cytoreductive surgery (CRS) plus hyperthermic intraperitoneal chemotherapy (HIPEC) for ovarian cancer. Methods: From April 2018 to November 2021, 66 patients with ovarian cancer were admitted to our hospital and randomly allocated to undergo intravenous chemotherapy following CRS (the observation group) or CRS with HIPEC (the experimental group) using a parallel randomized technique, with 33 cases in each group. Clinical effectiveness, intraoperative and postoperative recovery, VEGF level, T-lymphocyte subpopulation cell level, adverse events, and patient survival were all outcome metrics. Results: CRS plus HIPEC was associated with significantly higher clinical efficacy versus CRS alone (P < 0.05). The difference in the intraoperative bleeding and operative time between the two groups did not meet the statistical standard (P > 0.05). Patients in the experimental group experienced shorter postoperative chemotherapy and length of hospital stay than those in the observation group (P < 0.05). CRS plus HIPEC resulted in significantly lower levels of VEGFA, VEGFB, and VEGFC and higher levels of CD3+, CD4+, and CD3+/CD4+ than CRS alone (P < 0.05). The two groups of patients had a similar incidence of adverse events (P > 0.05). The experimental group showed a longer median survival (25 months) and a 1-year survival rate (79.55%) than the observation group (22 months, 49.56%) (log rank = 20.411, P < 0.05). A significantly lower 1-year recurrence rate was observed in the experimental group than in the observation group (P < 0.05). Conclusion: CRS plus HIPEC effectively improves the clinical efficacy of ovarian cancer patients, prolongs the survival of patients, and improves the level of VEGF and T-lymphocyte subpopulation cells, with a manageable safety. In addition, the treatment method can improve the therapeutic effect, reduce the toxic and side effects, and improve the immunity of the body, which is worthy of clinical promotion.

HRCT quantitative analysis of airway remodeling and airway trapping in the small airway asthma phenotype and its correlation with pulmonary function.

OBJECTIVES: We aimed to explore whether large airway remodeling and small airway structural changes exist in subjects with small airway asthma phenotype and to evaluate the relationships between quantitative high-resolution computed tomography (qHRCT) parameters and lung function. METHODS: We enrolled 15 subjects with small airway asthma phenotype and 18 healthy controls. The two groups were matched by age, sex and body square area (BSA) with propensity score matching (PSM). Pulmonary function and qHRCT parameters [wall thickness (WT), wall area (WA), lumen area (LA), wall area percentage (WA%) of the 4th-6th generations in the right upper lobe apical segmental bronchus (RB1), adjusted by BSA, WT/BSA, WA/BSA, and LA/BSA, relative volume change -860 HU to -950 HU (RVC-860 to -950) and the expiration to inspiration ratio of mean lung density (MLDE/I)) were compared between the groups. Correlation analysis was employed to assess the relationship between qHRCT parameters and pulmonary function. RESULTS: The small airway asthma phenotype had significantly higher WA%, RVC-860 to -950 and MLDE/I and lower LA/BSA than the healthy control. Additionally, we found moderate to strong correlations between impulse oscillation (IOS) indices and WA6% and WT6/BSA. No significant correlation was found between bronchial parameters and air trapping parameters (p > 0.05). CONCLUSIONS: Combining physiological tests with imaging approaches can lead to better evaluation of small airway disfunction (SAD) in asthmatic patients. Additionally, despite nonexistent airflow obstruction in patients with small airway asthma phenotype, large airway remodeling and small airway structural changes may appear simultaneously in the early stage of disease.

Short-acting beta-2 agonist prescription patterns and clinical outcomes in Chinese patients with asthma: an observational study in mainland China for the SABINA programme.

OBJECTIVES: The SABINA CHINA study aimed to determine prescription data for asthma medication with a focus on SABA and ICS in a representative population of patients with asthma in China. METHODS: SABINA China was a multicentre, observational, cross-sectional study with data collected retrospectively from a convenience sample of 25 tertiary centres across China. Patients (age ⩾ 12 years) with ⩾3 consultations/year were enrolled. Data were collected on clinical characteristics, asthma severity, and symptom control (as per GINA 2017), treatment and history of severe exacerbations over the past year. SABA over-prescription was defined as ⩾3 SABA canisters/year. Descriptive statistics are presented. RESULTS: Between March and August 2020, 498 patients were included in the outcome analysis. Mean (SD) age was 48.7 (15.0) years, 57.9% were female and 91% had moderate-to-severe asthma (n = 453). Overall, 12.5% (n = 62) and 26.4% (n = 131) of patients had uncontrolled and partly controlled asthma, respectively. SABA add-on was prescribed to 20.3% (n = 101) of patients; one patient with moderate-to-severe asthma was prescribed SABA-alone. SABA over-prescription in the overall population was 4.0% (n = 20; all with moderate-to-severe asthma) and 19.8% (20/101) among those prescribed SABA add-on. In the mild asthma group, 50% (n = 22) were prescribed ICS/LABA and 43.2% (n = 19) were prescribed LTRA. Among those with moderate-to-severe asthma, 97.4% (n = 441) were prescribed ICS/LABA and 55.0% (n = 249) were prescribed LTRA. Approximately 30% of patients (n = 149) experienced ⩾1% and 6.6% (n = 33) ⩾3 severe exacerbations in the preceding year; mean annual number of severe exacerbation/patient was 0.6 (1.2). Among those prescribed SABA add-on, ICS/LABA and LTRA (non-mutually exclusive groups due to overlapping prescriptions), 54.5%, 29.9%, and 35.3% had ⩾1 severe exacerbations, respectively. CONCLUSION: Among patients with predominantly moderate-to-severe asthma managed in tertiary care and were prescribed SABA, 1 in 5 received ⩾3 canisters/year. Fewer patients who received ICS/LABA prescriptions experienced annual exacerbations than those prescribed SABA add-on.

Clinical implications of concentration of alveolar nitric oxide in asthmatic and non-asthmatic subacute cough.

Asthma is an important cause of subacute cough. The concentration of alveolar nitric oxide (CANO) is a sensitive inflammatory indicator in peripheral airways, and it has received much less attention than the fraction of exhaled nitric oxide (FeNO50). The main objective of this study was to explore the correlation between CANO and clinical parameters in asthmatic and non-asthmatic subacute cough, which might promote understanding of the clinical utility of CANO in these special patient populations. 155 patients with subacute cough were included consecutively, of which 25 were diagnosed as asthmatic. Data for demographic characteristics, FeNO50, CANO, baseline spirometry, bronchial provocation test (or bronchodilation test) and response dose ratio (RDR) were collected. Differences between the asthmatic and non-asthmatic groups were analyzed. Spearman's correlation coefficient (ρ) was used to evaluate the correlation between FeNO50, CANO and other clinical parameters. In patients with subacute cough, baseline CANO values did not differ between asthmatic and non-asthmatic patients (4.4(1.3, 11.4) versus 4.0(2.1, 6.8) ppb,P> 0.05). Besides, CANO exhibited a stronger association with pulmonary function parameters when compared with FeNO50. For asthmatic subacute cough, CANO was inversely correlated with FEV1/FVC (ρ= -0.69,P< 0.01) and small airway parameters including MEF25 (ρ= -0.47,P< 0.05) and MMEF (ρ= -0.45,P< 0.05). For non-asthmatic subacute cough, CANO was inversely correlated with MEF25 (ρ= -0.19,P< 0.05) and RDR (ρ= -0.21,P< 0.05). In subacute cough, asthmatic and non-asthmatic patients had similar values of baseline CANO. In both asthmatic and non-asthmatic subacute cough, CANO exhibited a stronger association with pulmonary function parameters when compared with FeNO50. A low CANO value in non-asthmatic subacute cough corresponded to a higher value of RDR, which implied a stronger tendency towards airway responsiveness.

The Value of Impulse Oscillometric Parameters and Quantitative HRCT Parameters in Differentiating Asthma-COPD Overlap from COPD.

PURPOSE: To evaluate the value of impulse oscillometry (IOS) and quantitative HRCT parameters for differentiating asthma-COPD overlap (ACO) in COPD patients. PATIENTS AND METHODS: We enrolled 44 controls and 66 COPD patients, divided into the pure COPD group (n=40) and the ACO group (n=26). Spearman correlation analysis was utilized to assess the relationship between the quantitative HRCT and IOS parameters. A binary logistic regression analysis was performed to analyze the associations between the different variables and the risk of ACO. Receiver operating characteristic (ROC) curves were employed to identify the optimal cutoff and assess the diagnostic value of relative volume change -856 HU to -950 HU (RVC-856 to -950), decrease in the resistance from 5 Hz to 20 Hz (R5-R20) and their combination in predicting ACO. Bootstrapping validation was used to evaluate the internal validation. The concordance index (C-index) and calibration plot were calculated to assess the discrimination and calibration of the prediction model. RESULTS: Binary logistic regression analysis indicated that RVC-856 to -950 and the IOS parameters (R5-R20, R5, X5) were independently correlated with a higher risk of developing ACO after adjusting for age, sex, body mass index (BMI), history of smoking, exacerbation and atopy or allergic rhinitis. A correlation analysis showed a good correlation between the pulmonary function parameters and RVC-856 to -950, with a weaker correlation with the % area of low attenuation (LAA%) in ACO patients. Combining RVC-856 to -950 and R5-R20 to predict ACO, the AUC was 0.909, and the optimal cutoff value was >-0.62 for RVC-856 to -950 and >0.09 for R5-R20. Good calibration and favorable discrimination were displayed with a higher C-index. CONCLUSION: More serious small airway impairment exists in ACO patients. The combination of RVC-856 to -950 and R5-R20 could be applied to differentiate ACO from COPD patients.

Risk Factors of Difficult Pharyngeal Accidental Fishbones Ingestion.

OBJECTIVE: Accidental pharyngeal fishbone ingestion is a common complaint in ear, nose, and throat clinics. Approximately two-thirds of the accidentally ingested fishbones can be removed using tongue depressors and indirect laryngoscopy. However, the remaining third is challenging to identify and remove using these methods. These difficult fishbones require identification and removal via more advanced approaches. Video-guided laryngoscope is used to deal with difficult fishbones in our center. This study aimed to explore the risk factors for difficult fishbones. METHODS: A prospective study was performed at a teaching hospital on 2080 patients. Univariate and multivariate analyses were performed to identify the risk factors. RESULTS: The common fishbone locations were the tonsils (39.8%; defined as STEP-I), tongue base (37.1%), vallecula (13.3%; STEP-II), and hypopharynx (9.8%; STEP-III). With increasing STEP level, the ratio of difficult fishbones correspondingly increased (Z = 13.919, P < .001), and the proportions were 21.1%, 41.9%, and 70% in STEP-I, II, and III, respectively. In particular, fishbones in STEP-III (vs STEP-I) had a higher risk of difficult fishbones (odds ratio [OR]: 11.573, 95% CI: 7.987-16.769). Complaints of neck pain (yes vs no), foreign body sensation (yes vs no), and shorter length of fishbones always had a lower risk of difficult fishbones (OR: 0.455, 95% CI: 0.367-0.564; OR: 0.284, 95% CI: 0.191-0.422; OR: 0.727, 95% CI: 0.622-0.85). Missing teeth (yes vs no), swallowing behavior after fishbone ingestion (yes vs no), and male patients (vs female) had a higher risk of difficult fishbones (OR: 1.9, 95% CI: 1.47-2.456; OR: 1.631, 95% CI: 1.293-2.059; OR: 1.278, 95% CI: 1.047-1.56). CONCLUSIONS: Neck pain, foreign body sensation, fishbone length, patient age and sex, tooth status, and swallowing behavior after fishbone ingestion are independent risk factors for difficult fishbones.

The Value of Fractional Exhaled Nitric Oxide and Impulse Oscillometric and Spirometric Parameters for Predicting Bronchial Hyperresponsiveness in Adults with Chronic Cough.

PURPOSE: To evaluate the contribution of fractional exhaled nitric oxide (FeNO) and impulse oscillometry (IOS) and spirometric parameters in predicting bronchial hyperresponsiveness (BHR) in adults with chronic cough. PATIENTS AND METHODS: In total, 112 patients with chronic cough were enrolled in this prospective diagnostic study. Receiver operating characteristic (ROC) curves were generated to assess the diagnostic efficiency and optimal cut-off values of FeNO and IOS and spirometric parameters in predicting BHR. Optimal combinations of FeNO and IOS and spirometric parameters for BHR prediction were investigated using univariate and multivariate logistic regression models. Bootstrapping was employed for internal validation. Model discrimination and calibration were assessed using indices and calibration plots. RESULTS: Rhinitis and values of FeNO, IOS parameters (resonant frequency (Fres), reactance at 5 Hz (X5), and integrated area of low-frequency X (AX)) and spirometric parameters (FEV1, PEF, MEF75, MEF50, MEF25, MMEF) were significantly different between patients with BHR and those without BHR (P < 0.05). After adjusting for rhinitis, logistic analyses showed that FeNO combined with Fres, FeNO combined with MMEF, or the combination of FeNO, Fres and MMEF had high predictive value in diagnosing BHR; the areas under the ROC curves (AUCs) of the corresponding three models were 0.914, 0.919 and 0.927, respectively. In addition, the three models displayed good discrimination, with high C-index values and good calibration. CONCLUSION: FeNO combined with Fres or MMEF or a combination of these three parameters may be conveniently used as indicators in BHR prediction.

Value of Combining of the NLR and the Fibrinogen Level for Predicting Stroke-Associated Pneumonia.

PURPOSE: To evaluate the value of the NLR (neutrophil-to-lymphocyte ratio) and the fibrinogen level in predicting stroke-associated pneumonia (SAP) in acute ischemic stroke (AIS) patients. PATIENTS AND METHODS: In total, we enrolled 734 medical-ward patients with AIS in this retrospective study. Patients were divided into SAP (n=52) and non-SAP (n=682) groups according to the diagnostic criteria of SAP. Binary logistic regression analysis was used to analyze the relationship between the NLR, serum fibrinogen concentration and SAP. Receiver operating characteristic (ROC) curves were generated to identify the optimal cutoff points and assess the diagnostic value of the NLR, serum fibrinogen and the combination of NLR and fibrinogen in predicting SAP. RESULTS: SAP occurred in 52 (7.08%) patients among the enrolled AIS patients. Binary logistic regression analysis showed that the NLR (odds ratio [OR]: 2.802, 95% confidence interval [CI]: 1.302-6.032, P=0.008) and serum fibrinogen concentration (OR: 7.850, 95% CI: 3.636-16.949, P=0.000) were independently associated with a higher risk of SAP incidence after adjusting for age, sex, ASPECT score, atrial fibrillation, nasogastric tube feeding, LDL-C and TC, temperature at admission and mechanical ventilation. The optimal cutoff points of the NLR and serum fibrinogen to distinguish SAP among AIS patients were 3.603 (AUC, 0.690; NPV, 95.78; PPV, 19.01) and 4.595 (AUC, 0.727; NPV, 95.60; PPV, 24.49), respectively. When the combination of NLR and fibrinogen was used to predict SAP, the optimal cutoff points were >2.436 for NLR and >3.24 for fibrinogen (AUC, 0.758; NPV, 98.50; PPV, 11.80). CONCLUSION: The NLR and serum fibrinogen might have greater negative diagnostic value in predicting SAP among AIS patients. Combining the NLR and serum fibrinogen showed an increased AUC for predicting SAP among AIS patients.

Diagnostic value of FeNO and MMEF for predicting cough variant asthma in chronic cough patients with or without allergic rhinitis.

OBJECTIVE: To evaluate the diagnostic value of fractional exhaled nitric oxide (FeNO) and maximum mid-expiratory flow (MMEF) for differentiating cough variant asthma (CVA) from chronic cough in patients with or without allergic rhinitis. METHODS: In total, 328 patients with chronic cough who underwent spirometry and FeNO testing were consecutively included in the retrospective analysis. Patients were divided into the CVA (n = 125) or NCVA (n = 203) groups according to the diagnostic criteria of CVA. Receiver operating characteristic (ROC) curves were established to assess the diagnostic efficiency and optimal cutoff points of FeNO and MMEF for the prediction of CVA. RESULTS: The optimal cutoff values of FeNO and MMEF to discriminate CVA from chronic cough were 24.5 ppb (AUC, 0.765; sensitivity, 69.60%; specificity 72.91%; PPV, 61.27%; NPV, 79.57%) and 66.2% (AUC, 0.771; sensitivity, 67.20%; specificity 78.33%; PPV, 65.63%; NPV, 79.50%). The optimal cutoff values of combining FeNO with MMEF to discriminate CVA from chronic cough were >22 ppb for FeNO and <62.6% for MMEF (AUC, 0.877). In patients with and without allergic rhinitis, the optimal cutoff point of FeNO to discriminate CVA from chronic cough was 24.5 ppb (AUC, 0.820) and 33.5 ppb (AUC, 0.707), respectively. CONCLUSIONS: FeNO and MMEF might have greater value as negative parameters for differentiating CVA from chronic cough. Combining FeNO and MMEF provided a significantly better prediction than either alone. The diagnostic accuracy of FeNO for predicting CVA in chronic cough patients with allergic rhinitis was higher than in chronic cough patients without allergic rhinitis.

Assessment of Internal Carotid Artery Invasion With the Endoscopic Endonasal Approach: Implications of a New Grading System and Security Strategy.

BACKGROUND: Internal carotid artery (ICA) injury during the endoscopic endonasal approach (EEA) for ventral cranial base tumors is rare but fatal. OBJECTIVE: The aim of this study was to investigate the relationship between ventral cranial base tumors and corresponding ICA using quantitative means to improve the safety of surgery. MATERIAL AND METHODS: In this retrospective study, a total of 46 patients with ventral cranial base tumors undergoing surgery with the EEA were enrolled. Preoperative imaging features and clinical data were recorded. The influencing factors of the gross total resection (GTR) rate were analyzed. A new grading system and security strategy were developed. RESULTS: Postoperative enhanced magnetic resonance imaging showed a GTR of 61.7%. There was a significant correlation between the GTR rate and tumors surrounding the ICA, the tumor site, the involved ICA segment, and the blood supply (P < 0.05). We included preoperative radiotherapy as supplementary data in the grading system. Each variable involved the letter "S"; (surround, site, segment, blood supply, supplementary data); hence, the system was named the 5S ICA grading scale. CONCLUSION: We recommend a safety assessment through the 5S ICA grading scale and the ICA grading strategy to obtain the maximum total resection rate while ensuring patient safety.

Pleomorphic adenoma of the trachea: A case report and review of the literature.

BACKGROUND: Pleomorphic adenoma (PA) is the most common benign tumor that occurs in the salivary glands; however, tracheobronchial PA is rarely observed. To the best of our knowledge, fewer than 50 cases have been reported in the literature. We report a 49-year-old woman who had been treated for asthma for 2 years before being diagnosed with PA of the trachea. CASE SUMMARY: A 49-year-old woman was referred to our hospital due to dyspnea upon exertion and chronic cough with wheezing for 2 years. Laboratory tests showed an elevated white blood cell count, absolute neutrophil count, and percentage of neutrophils. A chest computerized tomography scan showed a well-defined, soft-tissue density lesion measuring 2.4 cm × 2.1 cm in the lower trachea. Flexible bronchoscopy revealed that nearly 90% of the tracheal lumen was obstructed. The histopathological and immunohistochemistry features suggested PA of the trachea. Furthermore, we review the characteristics of 29 patients with tracheobronchial PA over the last 30 years. CONCLUSION: Tracheobronchial PA occurs without gender predominance, mostly in the lower or upper trachea, and has a low recurrence rate. The median age at diagnosis is 48 years. The most common symptoms are cough, stridor, dyspnea, and wheezing.

Abl and Canoe/Afadin mediate mechanotransduction at tricellular junctions.

Epithelial structure is generated by the dynamic reorganization of cells in response to mechanical forces. Adherens junctions transmit forces between cells, but how cells sense and respond to these forces in vivo is not well understood. We identify a mechanotransduction pathway involving the Abl tyrosine kinase and Canoe/Afadin that stabilizes cell adhesion under tension at tricellular junctions in the Drosophila embryo. Canoe is recruited to tricellular junctions in response to actomyosin contractility, and this mechanosensitivity requires Abl-dependent phosphorylation of a conserved tyrosine in the Canoe actin-binding domain. Preventing Canoe tyrosine phosphorylation destabilizes tricellular adhesion, and anchoring Canoe at tricellular junctions independently of mechanical inputs aberrantly stabilizes adhesion, arresting cell rearrangement. These results identify a force-responsive mechanism that stabilizes tricellular adhesion under tension during epithelial remodeling.

Tissue microarray analysis reveals that cofilin expression is a poor prognostic factor in juvenile nasopharyngeal angiofibroma.

BACKGROUND: Juvenile nasopharyngeal angiofibroma (JNA) has a high recurrence rate after surgery. Cofilin overexpression is associated with increased tumor cell metastasis, and progression of various human cancers. However, studies on cofilin expression in JNA are rare. The purpose of this study was to investigate the expression and localization of cofilin in a tissue microarray (TMA) of JNA specimens. In addition, we also analyzed its correlation with clinicopathological features and recurrence. METHODS: Immunohistochemistry was performed to detect cofilin expression in a TMA of samples from 70 JNA patients and 10 control subjects. The association between clinicopathological variables and cofilin immunostaining was analyzed using Pearson's chi-square test. Kaplan-Meier survival analysis was used to calculate the disease-free survival rate, and investigate the effect of cofilin expression on time to recurrence (TTR) in JNA patients. The Cox regression model was used for multivariate survival analysis. RESULTS: Cofilin was detected in irregular smooth muscle cells, pericytes, less differentiated stromal cells, and plump cells, but not in inactive fibroblasts and mature vascular endothelial cells of JNA specimens. The presence of cofilin in JNA was correlated with tumor stage (p = 0.012) and volume of intraoperative hemorrhage (p < 0.001). JNA patients with high cofilin expression had a higher recurrence rate than those with low cofilin expression (p = 0.012). Cofilin expression and patient's age were significant predictors of TTR, and cofilin was a better predictor for disease recurrence (area under the receiver operating curve [AUROC; 0.711; p = 0.005) than other clinicopathological features. CONCLUSION: Cofilin is an independent prognostic marker for JNA patients who have undergone surgical treatment and may represent a novel therapeutic target for extensive JNA.

Study on the Mechanism of Curcumin Regulating Lung Injury Induced by Outdoor Fine Particulate Matter (PM2.5).

BACKGROUND: Epidemiological studies have shown that exposure to PM induces oxidative stress, leading to a variety of health problems. In particular, PM2.5 contains a lot of substances harmful to the human body and penetrates into the lungs to induce lung injury. At the same time, there is increasing evidence that oxidative stress also affects the severity of lung injury. However, there is still no good way to reduce or eliminate these hazards. In the future, more experimental research is needed to further confirm the mechanisms of these hazards and formulate effective preventive measures and treatment plans for their hazard mechanisms. Curcumin has been reported to reduce oxidative stress and inflammatory damage and protect organs. OBJECTIVE: To investigate whether curcumin can play a protective role against PM2.5-induced oxidative stress and inflammatory damage by inducing expression of the HO-1/CO/P38 MAPK pathway. METHODS: In this experiment, PM2.5 was dropped into the trachea to establish a lung injury model in mice. 28 SPF-grade male Kunming mice were randomly divided into 4 groups: normal control group, saline control group, PM2.5 treatment group, and curcumin intervention group. Albumin (ALB), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) were measured in alveolar lavage fluid (BALF) to assess lung tissue damage. Colorimetric detection of oxidative stress indicators such as MDA, GSH-PX, T-AOC, and CAT in the lung tissue was performed. The levels of IL-6 and TNF-α in the lung tissue were determined by ELISA. Histopathological examination was used for the assessment of alveolar epithelial damage. The protein expression of the HO-1/P38 MAPK pathway in the lung tissue was determined by Western blot and immunohistochemistry. Endogenous CO was detected by spectrophotometry. The results showed that the expression of the HO-1/CO/P38 MAPK protein in the lung tissue was significantly increased in the curcumin intervention group compared with the PM2.5 treatment group, and it was statistically significant (P < 0.05). Compared with the PM2.5 treatment group, the curcumin intervention group can reduce the amount of ALB, LDH, and ALP in BALF; reduce the levels of MDA, IL-1, and TNF-α in the lung tissue; and improve GSH-PX, T-AOC, and CAT levels, but there is no statistical difference (P > 0.05). CONCLUSION: We found that PM2.5 can cause lung damage through oxidative stress and inflammatory responses. Oxidative stress and inflammatory responses increase the expression of HO-1/CO/P38 MAPK. The intervention of curcumin can further increase the expression of HO-1/CO/P38 MAPK.

Low carotid endothelial shear stress associated with cerebral small vessel disease in an older population: A subgroup analysis of a population-based prospective cohort study.

BACKGROUND AND AIMS: The association between carotid wall shear stress (WSS) and cerebral small vessel disease has yet to be fully elucidated. The major purpose of this study was to investigate this association in older subjects. METHODS: Common carotid artery WSS, endothelial function, white matter hyperintensities (WMH), lacunes, and microbleeds were assessed in 1396 older adults. Participants were followed-up for an average of 69.7 months. RESULTS: Mean (M) and peak (P) WSS and changes in endothelial function were independently associated with changes in WMH volume and fraction, lacune counts, and microbleed counts (all p < 0.05). The risks of new-incident Fazekas scale ≥2 [hazard ratio (HR) with 95% confidence interval (CI): 2.141 (1.469-3.119), p = 0.005 and 1.731 (1.197-2.505), p = 0.004, respectively], lacunes [HR (95% CI): 2.034 (1.369-3.022), p < 0.001 and 1.693 (1.151-2.490), p = 0.003, respectively], and microbleeds [HR (95% CI): 2.311 (1.509-3.541), p < 0.001 and 2.208 (1.299-3.751), p < 0.001, respectively] were significantly higher in the lowest quartile group than in the higher quartile group, as classified by either MWSS or PWSS, after adjustment for confounders. CONCLUSIONS: Low carotid WSS is an independent risk factor for the progression of cerebral small vessel disease in older adults.

Epithelial­mesenchymal transition in chronic rhinosinusitis (CRS) and the prognostic value of α­SMA in postoperative outcomes of patients with CRS.

Tissue remodeling is the pathological basis of the symptoms encountered in chronic rhinosinusitis (CRS). Epithelial­mesenchymal transition (EMT) may participate in this process. The present study was designed to investigate the involvement of EMT in CRS. In addition, the prognostic value of the EMT biomarker α­smooth muscle actin (α­SMA) was assessed in patients with CRS who underwent endoscopic sinus surgery (ESS). A total of 13 patients with CRS without nasal polyps (CRSsNP), 13 patients with CRS with nasal polyps (CRSwNP) and 13 control subjects were enrolled. The expression of EMT markers was determined in sinonasal specimens by qPCR, western blot and immunofluorescence assays. EMT features were evaluated in primary nasal epithelial cells (NECs) with transforming growth factor (TGF)­ß1 stimulation. The associations were assessed between α­SMA expression and the clinical features of CRS. Epithelial and mesenchymal markers were overexpressed in the sinonasal specimens of both CRSsNP and CRSwNP patients. Alterations in the expression pattern were more apparent in the CRSsNP patients. Following incubation of primary NECs with TGF­ß1, a mesenchymal shape was acquired. In addition, NECs that co­expressed α­SMA and cytokeratin were readily detected and the protein levels of α­SMA were elevated. In contrast to α­SMA, the levels of E­cadherin were decreased. The protein levels of α­SMA were negatively correlated with endoscopic scores and several postoperative symptoms. In conclusion, partial EMT occurred in patients with CRS, notably in CRSsNP patients. Moreover, primary NECs could undergo EMT following TGF­ß1 treatment in vitro. In addition, α­SMA could be considered an efficient predictor for postoperative endoscopic and symptomatic outcomes in patients with CRS treated with ESS.

TGF­ß1 upregulates the expression of lncRNA­ATB to promote atherosclerosis.

Transforming growth factor (TGF)­ß1 is reported to be associated with the occurrence of atherosclerosis, although the mechanism remains unclear. Therefore, the present study aimed to investigate the involvement of TGF­ß1 signaling in atherosclerosis. A total of 56 patients with atherosclerosis and 44 healthy volunteers were involved in this study. Serum expression of TGF­ß1 and long non­coding RNA­ATB was detected by ELISA and quantitative polymerase chain reaction (qPCR). Receiver operating characteristic curve analysis was performed to analyze the diagnostic value of serum TGF­ß1 and lncRNA­ATB for atherosclerosis. A human umbilical vein endothelial cell (HUVEC) line overexpressing lncRNA­ATB was constructed. The effects of TGF­ß1 treatment and lncRNA­ATB overexpression on HUVEC cell proliferation and viability was detected with Cell Counting Kit­8 and MTT assays, respectively. Expression of TGF­ß1 and pro­apoptotic Caspase­3 in lncRNA­ATB­overexpressing HUVECs was detected by western blotting. In addition, the expression of lncRNA­ATB in TGF­ß1­treated HUVECs was detected by qPCR. It was demonstrated that serum TGF­ß1 and lncRNA­ATB expression was significantly higher in atherosclerosis patients, compared with controls, and could be used to effectively distinguish patients from healthy individuals. TGF­ß1 treatment and lncRNA­ATB overexpression reduced HUVEC viability and proliferation. TGF­ß1 treatment increased the expression of lncRNA­ATB in HUVECs, while lncRNA­ATB overexpression had no significant effect on TGF­ß1 expression. LncRNA­ATB silencing with small interfering RNA significantly reduced the effects of TGF­ß1 treatment on the proliferation and viability of HUVECs. Furthermore, LncRNA­ATB overexpression upregulated the expression of caspase­3 in HUVECs. Therefore, it was concluded that TGF­ß1 may have upregulated the expression of lncRNA­ATB to promote atherosclerosis, and lncRNA­ATB may serve as a potential therapeutic target for atherosclerosis. However, the mechanism remains to be further investigated.