Prognostic value of serum oncomarkers for patients hospitalized with acute exacerbation of interstitial lung disease.

BACKGROUND: Different types of inflammatory processes and fibrosis have been implicated in the pathogenesis of interstitial lung disease (ILD), a heterogeneous, diffuse, parenchymal lung disease. Acute exacerbation (AE) of ILD is characterized by significant respiratory deterioration and is associated with high mortality rates. Several serum oncomarkers have been used to determine the prognosis of ILD; however, the prognostic value of serum oncomarker levels in patients with AE-ILD remains unclear. OBJECTIVE: To evaluate the prognostic value of serum oncomarker levels in patients with AE-ILD and its main subtypes. DESIGN: Retrospective study. METHODS: The serum levels of 8 oncomarkers in 281 patients hospitalized with AE-ILD at our institution between 2017 and 2022 were retrospectively reviewed. The baseline characteristics and serum oncomarker levels were compared between the survival and non-survival groups of AE-ILD and its main subtypes. Multivariate logistic regression analysis was performed to identify independent prognosis-related markers, and the best prognostic predictor was analyzed using receiver operating characteristic curve (ROC) analysis. RESULT: Idiopathic pulmonary fibrosis (IPF; n = 65), idiopathic nonspecific interstitial pneumonia (iNSIP; n = 26), and connective tissue disease-associated interstitial lung disease (CTD-ILD; n = 161) were the three main subtypes of ILD. The in-hospital mortality rate among patients with AE-ILD was 21%. The serum oncomarker levels of most patients with AE-ILD and its main subtypes in the non-survival group were higher than those in the survival group. Multivariate analysis revealed that ferritin and cytokeratin 19 fragments (CYFRA21-1) were independent prognostic risk factors for patients hospitalized with AE-ILD or AE-CTD-ILD. CYFRA21-1 was identified as an independent prognostic risk factor for patients hospitalized with AE-IPF or AE-iNSIP. CONCLUSION: CYFRA21-1 may be a viable biomarker for predicting the prognosis of patients with AE-ILD, regardless of the underlying subtype of ILD. Ferritin has a prognostic value in patients with AE-ILD or AE-CTD-ILD.

Clinical manifestations and prognostic factors analysis of patients hospitalised with acute exacerbation of idiopathic pulmonary fibrosis and other interstitial lung diseases.

BACKGROUND: Acute exacerbation (AE) is a life-threatening condition taking place not only in idiopathic pulmonary fibrosis (IPF) but also in interstitial lung diseases (ILD) other than IPF (non-IPF ILD). This study aims to compare the clinical manifestations between patients hospitalised with AE-IPF and AE-non-IPF ILD, and further analyse the risk factors related to in-hospital mortality. METHODS: Clinical data of 406 patients hospitalised with AE-IPF (93 cases) and AE-non-IPF ILD (313 cases) were retrospectively collected. Clinical features were compared between the two groups. Risk factors related to in-hospital mortality in patients with overall AE-ILD, AE-IPF and AE-non-IPF ILD were identified by multiple logistic regression analyses, respectively, and assessed by receiver operating characteristic curve. RESULTS: In addition to having more smokers and males, the AE-IPF group also had more respiratory failure on admission, comorbidities of pulmonary hypertension (PAH) or coronary artery disease/heart failure, a longer history of pre-existing ILD. Comorbidity of coronary heart disease/heart failure, respiratory failure at admission, neutrophil (N)%, serum hydroxybutyrate dehydrogenase (HBDH), lactate dehydrogenase (LDH) and low cholesterol levels were independent risk factors for patients with AE-ILD, while respiratory failure on admission, N%, serum HBDH, urea nitrogen, LDH and low albumin levels were risk factors for the AE-non-IPF ILD group, and fever, N% and PAH were the AE-IPF group's. Among them, HBDH 0.758 (sensitivity 85.5%, specificity 56%, cut-off 237.5 U/L) for patients with AE-ILD; N% 0.838 (sensitivity 62.5%, specificity 91.18%, cut-off 83.55%) for the AE-IPF group and HBDH 0.779 (sensitivity 86.4%, specificity 55.1%, cut-off 243.5 U/L) for the AE-non-IPF ILD group were the risk factors with the highest area under the curve. CONCLUSIONS: Clinical characteristics differ between patients with AE-IPF and AE-non-IPF ILD. HBDH outperformed LDH in predicting the prognosis for patients with AE-ILD and AE-non-IPF ILD. N% was an independent predictor of death in-hospital in all three groups, especially in the AE-IPF group.

Glucocorticoid Use in Patients Hospitalized with Chronic Obstructive Pulmonary Disease Exacerbations.

Objective of the Study: Systemic glucocorticoid therapy can improve the outcomes of acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The study tried to investigate the use of glucocorticoids in AECOPD patients and the factors associated with the physicians' choice. Methodology: Patients with AECOPD over two periods were divided by the year of 2017 when GOLD and ERS/ATS Guideline for COPD were updated. Data of patients regarding the study was retrieved from medical records. Descriptive statistical analysis was used for the illustration of glucocorticoids use, and hypothesis testing for comparison over the periods. Results: Between 2010 and 2016, the proportion of ICS use was 522/640 (81.6%) and 341/452 (75.4%) between 2017 and 2020. COPD severity (GOLD C/D classification), bronchial asthma, percentage of neutrophils, and higher PaCO2 were factors associated with physicians' prescription of systemic glucocorticoids between 2010 and 2016. While the use of ICS at the stable stage, counts of neutrophils, and higher PaCO2 were influencing factors between 2017 and 2020. Over the two periods, 1-year recurrent rate decreased from 32.4% to 20.9%, with a significant statistical difference (P<0.001). Conclusion: The optimized use of glucocorticoids was found after the publishment of 2017 ERS/ATS Guideline for COPD, this improvement was associated with a decreased 1-year recurrence rate among AECOPD patients at our institution, underscoring the positive impact of guideline updates on patient outcomes.

[Multiple pulmonary nodules with interstitial changes].

A 53-year-old female patient with pulmonary nodules for more than 3 years was admitted to Beijing Chao-Yang Hospital because of cough and sputum with shortness of breath after exercise for 4 months. In the first two and a half years, her pulmonary nodules remained stable, after that the nodules increased obviously with interstitial changes. After admission, a venous thromboembolic (VTE) event was quickly detected with a marked increase in D-dimer. Then, based on the clues of VTE examination, bronchoscopy, gastroscope, positron emission tomography-CT, head magnetic resonance and other examinations were performed. The final pathological diagnosis was lung adenocarcinoma, mainly solid with mucus secretion, with mediastinal hilar lymph node metastasis, intrapulmonary metastasis and gastric metastasis. Gene detection of lung and stomach histopathological tissues showed positive EML4-ALK fusion gene. The patient received therapies with crizotinib, alectinib in sequence and anticoagulation. After 20-month treatment, a telephone follow-up showed that there was no significant limitation in her daily activities.

Reticulocalbin 3 deficiency in alveolar epithelium attenuated LPS-induced ALI via NF-κB signaling.

Acute respiratory distress syndrome (ARDS) is characterized by acute lung injury (ALI) secondary to an excessive alveolar inflammatory response. Reticulocalbin 3 (Rcn3) is an endoplasmic reticulum (ER) lumen protein in the secretory pathway. We previously reported the indispensable role of Rcn3 in type II alveolar epithelial cells (AECIIs) during lung development and the lung injury repair process. In the present study, we further observed a marked induction of Rcn3 in the alveolar epithelium during LPS-induced ALI. In vitro alveolar epithelial (MLE-12) cells consistently exhibited a significant induction of Rcn3 accompanied with NF-κB activation in response to LPS exposure. We examined the role of Rcn3 in the alveolar inflammatory response by using mice with a selective deletion of Rcn3 in alveolar epithelial cells upon doxycycline administration. The Rcn3 deficiency significantly blunted the ALI and alveolar inflammation induced by intratracheal LPS instillation but not that induced by an intraperitoneal LPS injection (secondary insult); the alleviated ALI was accompanied by decreases in NF-κB activation and NLRP3 levels but not in GRP78 and cleaved caspase-3 levels. The studies conducted in MLE-12 cells consistently showed that Rcn3 knockdown blunted the activations of NF-κB signaling and NLRP3-dependent inflammasome upon LPS exposure. Collectively, these findings suggest a novel role for Rcn3 in regulating the alveolar inflammatory response to pulmonary infection via the NF-κB/NLRP3/inflammasome axis and shed additional light on the mechanism of ARDS/ALI.

Rcn3 Suppression Was Responsible for Partial Relief of Emphysema as Shown by Specific Type II Alveolar Epithelial Cell Rcn3 CKO Mouse Model.

BACKGROUND: Chronic obstructive pulmonary disease (COPD), characterized by irreversible airflow limitation, is a highly prevalent lung disease worldwide and imposes increasing disease burdens globally. Emphysema is one of the primary pathological features contributing to the irreversible decline of pulmonary function in COPD patients, but the pathogenetic mechanisms remain unclear. Reticulocalbin 3 (Rcn3) is an endoplasmic reticulum (ER) lumen protein localized in the secretory pathway of living cells. Rcn3 in type II alveolar epithelial cell (AECIIs) has been reported to play a critical role in regulating perinatal lung development and bleomycin-induced lung injury-repair processes. We hypothesized that Rcn3 deficiency is associated with the development of emphysema during COPD, which is associated with the dysfunction of injury-repair modulated by alveolar epithelial cells. MATERIALS AND METHODS: We examined Rcn3 expression in lung specimens from COPD patients and non-COPD control patients undergoing lung lobectomy or pneumonectomy. Two mouse models of emphysema were established by cigarette smoke (CS) exposure and intratracheal instillation of porcine pancreatic elastase (PPE). Rcn3 expression was detected in the lung tissues from these mice. Furthermore, conditional knockout (CKO) mice with Rcn3 deletion specific to AECIIs were used to explore the role of Rcn3 in PPE-induced emphysema progression. Rcn3 protein expression in lung tissues was evaluated by Western blot and immunohistochemistry. Rcn3 mRNA expression in lung tissues was detected by qPCR. RESULTS: Rcn3 expression was significantly increased in the lung specimens from COPD patients versus non-COPD patients and the level of Rcn3 increase was associated with the degree of emphysema. Rcn3 expression were also significantly up-regulated in both CS-induced and PPE-induced emphysematous mouse lungs. Moreover, the selective ablation of Rcn3 in AECIIs significantly alleviated severity of the mouse emphysema in response to intratracheal installation of PPE. CONCLUSION: Our data, for the first time, indicated that suppression of Rcn3 expression in AECIIs has a beneficial effect on PPE-induced emphysema.

Blood Eosinophil Levels and Prognosis of Hospitalized Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease.

BACKGROUD: Studies about the clinical significance of high eosinophil levels in chronic obstructive pulmonary disease (COPD) are conflicting, and it has been less studied in hospitalized patients with acute exacerbation of COPD (AECOPD).This study was to examine blood eosinophil levels in relation to the prognosis of hospitalized patients with AECOPD. METHODS: This was a retrospective cohort study of patients with AECOPD as their primary diagnosis and admitted to Beijing Shijitan Hospital, Capital Medical University, from January 2010 to December 2016. The patients were assigned according to the count of eosinophil in peripheral blood at their first hospitalization. Patients were grouped as ≤100, 100-300, and ≥300 eosinophils/µL of peripheral blood. The use of glucocorticoids, duration of hospitalization, in-hospital mortality, and re-hospitalization were examined. RESULTS: Compared with the 100-300 eosinophils/µL group, the ≤100 eosinophils/µL group showed higher frequencies of fever, respiratory failure, and the use of systemic glucocorticoids. Eosinophil counts were not associated with in-hospital mortality and duration of hospitalization. The multivariable analysis showed that GOLD3/4 (odds ratio (OR)=2.04, 95%CI: 1.20-3.44, P = 0.008), systemic glucocorticoids (OR=1.84, 95%CI: 1.41-2.98, P = 0.012), mechanical ventilation (OR=2.66, 95%CI: 1.36-5.18, P = 0.004), and acute exacerbation in the past year before hospitalization (OR=2.03, 95%CI: 1.27-3.23, P = 0.003) were independently associated with acute exacerbation within 1 year after discharge. Eosinophil count was not associated with acute exacerbation within 1 year after discharge. CONCLUSIONS: Peripheral blood eosinophil counts are not associated with the 1-year AECOPD prognosis.

Independent factors associated with pneumonia among hospitalized patients with acute exacerbations of chronic obstructive pulmonary disease.

Acute exacerbations (AE) affect the prognosis of hospitalized patients with chronic obstructive pulmonary disease (COPD). Pneumonia further affects their prognosis and early diagnosis of pneumonia in AECOPD is important to initiate treatments. This study aimed to examine the differences between hospitalized AECOPD patients with and without pneumonia in order to identify risk factors of pneumonia among hospitalized patients with AECOPD.This was a retrospective case-control study of patients with COPD hospitalized at the respiratory ward of Beijing Shijitan Hospital, Capital Medical University, from October 2010 to October 2013. Patients were divided into the pneumonia and nonpneumonia groups based on exudations or opacities on chest computed tomography (CT) at admission. Data were analyzed using the chi-square test and independent 2-sample ANOVA in SPSS 20.0. Logistic regression analysis was used to identify the factors independently associated with pneumonia. P < .05 was considered statistically significant.A total of 164 patients were included. Smoking history (OR = 2.646, 95%CI 1.153-6.074, P = .022), use of drugs during the stable stage (OR = 0.435, 95%CI 0.216-0.877, P = .020), D-dimer levels (OR = 1.001, 95%CI 1.000-1.002, P = .049), percentage of neutrophils (OR = 0.271, 95%CI 0.078-0.940, P = .040), and magnitude of neutrophils increase (OR = 0.946, 95%CI 0.896-0.999, P = .046) were independently associated with pneumonia in patients with AECOPD. For severe and very severe COPD patients, smoking history (OR = 4.426, 95%CI 1.458-13.435, P = .009), use of drugs during the stable stage (OR = 0.384, 95%CI 0.168-0.877, P = .042), and fever (OR = 0.426, 95%CI 0.187-0.969, P = .023) were independently associated with pneumonia.Smoking history, use of drugs during the stable stage, and percentage of neutrophils are independently associated with CT-diagnosed pneumonia among hospitalized AECOPD patients.

Dietary Supplementation with Oleum Cinnamomi Improves Intestinal Functions in Piglets.

The present study was to determine the efficacy of dietary supplementation with oleum cinnamomi (OCM) on growth performance and intestinal functions in piglets. Sixteen piglets (24-day-old) were randomly assigned to the control or OCM groups. Piglets in the control group were fed a basal diet, whereas piglets in the OCM group were fed the basal diet supplemented with 50 mg/kg OCM. On day 20 of the trial, blood samples and intestinal tissues were obtained from piglets. Compared with the control group, dietary OCM supplementation increased (p < 0.05) average daily feed intake, plasma insulin levels, villus width and villous surface area in the duodenum and jejunum, DNA levels and RNA/DNA ratios in the ileum, the abundance of Enterococcus genus and Lactobacillus genus in caecum digesta, mRNA levels for epithelial growth factor receptor (EGFR), Ras, extracellular signal-regulated kinase 1/2 (Erk1/2), b-cell lymphoma-extra large (Bcl-xL), villin, junctional adhesion molecule A (JAM-A), myxovirus resistance (MX) 1, MX2 and regenerating islet-derived protein 3 gamma (REG3G), and protein abundances of Ras and claudin-1, but decreased (p < 0.05) diarrhoea incidence; the abundances of Enterobacteriaceae family, Enterococcus genus, Lactobacillus genus, Bifidobacterium genus, and Clostrium coccoides in the colon digesta, and AMP-activated protein kinase (AMPK) mRNA levels and caspase-3 protein abundance in the jejunal mucosa of piglets. Taken together, these data indicate that dietary OCM supplementation modulates intestinal microbiota and improves intestinal function in weanling pigs. OCM is an effective feed additive and alternative to feed antibiotics for improving intestinal health in swine.

Reticulocalbin 3 Deficiency in Alveolar Epithelium Exacerbated Bleomycin-induced Pulmonary Fibrosis.

Reticulocalbin 3 (Rcn3) is an endoplasmic reticulum (ER) lumen protein localized to the secretory pathway. We have reported that Rcn3 plays a critical role in alveolar epithelial type II cell maturation during perinatal lung development, but its biological role in the adult lung is largely unknown. In this study, we found marked induction of Rcn3 expression in alveolar epithelium during bleomycin-induced pulmonary fibrosis, which is most obvious in alveolar epithelial type II cells (AECIIs). To further examine Rcn3 in pulmonary injury remodeling, we generated transgenic mice to selectively delete Rcn3 in AECIIs in adulthood. Although Rcn3 deletion did not cause obvious abnormalities in the lung architecture and mechanics, the exposure of Rcn3-deleted mice to bleomycin led to exacerbated pulmonary fibrosis and reduced lung mechanics. These Rcn3-deleted mice also displayed enhanced alveolar epithelial cell (AEC) apoptosis and ER stress after bleomycin treatment, which was confirmed by in vitro studies both in primary AECIIs and mouse lung epithelial cells. Consistently, Rcn3 deficiency also enhanced ER stress and apoptosis induced by ER stress inducers, tunicamycin and thapsigargin. In addition, Rcn3 deficiency caused blunted wound closure capability of AECs, but not altered proliferation and bleomycin-induced epithelial-mesenchymal transition process. Collectively, these findings indicate that bleomycin-induced upregulation of Rcn3 in AECIIs appears to contribute to AECII survival and wound healing. These observations, for the first time, suggest a novel role of Rcn3 in regulating pulmonary injury remodeling, and shed additional light on the mechanism of idiopathic pulmonary fibrosis.

Clinical characteristics of patients with chronic obstructive pulmonary disease overlapped with bronchial asthma.

BACKGROUND: The clinical characteristics of patients with chronic obstructive pulmonary disease overlapped with bronchial asthma (COPD-BA) have not been discussed thoroughly. OBJECTIVE: To reveal the clinical features of patients with COPD-BA, to evaluate the risk factors of COPD-BA, and to provide suggestions for COPD individualized therapy. METHODS: A retrospective observational study was performed. A total of 182 patients with COPD (90 with COPD-BA and 92 with pure COPD) were recruited in the study. Information on the following items was collected: demographics, clinical manifestations, complications, laboratory findings, other histories, and inpatient treatments during exacerbation. RESULTS: A total of 182 patients were diagnosed with COPD, with 90 (49.45%) being classified as having COPD-BA. Patients with COPD-BA were more likely to be female (P = .004) and experienced more severe respiratory exacerbations (P = .04) despite being younger (P = .008). Those patients at onset of recurrent cough and sputum production were younger (P = .001). Significantly, a positive asthmatic family history (P = .03) was observed. Patients with COPD-BA usually had higher level of total serum IgE (although no differences were observed), had higher positive rates of the serum specific IgE (P = .004), and were more like to have an allergic history (P = .003). Allergic factor was the risk factor of COPD-BA (odds ratio, 4.477). During hospitalization, patients with COPD-BA tended to be treated with systemic corticosteroids (P = .008). CONCLUSION: Patients with COPD-BA were characterized by persistent airflow limitation with unique clinical features. Allergic factor was associated with the presence of asthmatic characteristics in patients with COPD. When hospitalized for exacerbation, the individualized therapy for COPD-BA might include the use of corticosteroids systemically.

Disseminated nocardiosis caused by Nocardia otitidiscaviarum in an immunocompetent host: A case report and literature review.

The aim of the present study was to summarize the clinical characteristics of nocardiosis caused by Nocardia otitidiscaviarum in order to improve the knowledge of nocardiosis. A case of dissemination nocardiosis caused by N. otitidiscaviarum in an immunocompetent host is reported and the associated literature reviewed. Informed consent for publication of this case report was provided by the patient. The present patient was a young immunocompetent man suffering from disseminated nocardiosis induced by infection with N. otitidiscaviarum. Following a poor response to ß-lactam antibiotic, a combination of sulfonamide with minocycline was administered, which successfully ameliorated the symptoms. Previous studies published in English were retrieved from PubMed with 'Nocardia otitidiscaviarum' used as the search keyword. A total of 23 articles were retrieved from the PubMed database, supporting the assertion that N. otitidiscaviarum is a rare Nocardia species. Among these 23 cases, there were 11 cases of lymphocutaneous (48%), 5 of pulmonary (22%), 2 of brain (9%) and 1 of pyothorax (4%) infection, and 4 cases of disseminated infections (17%). Analysis of the immune state of these patients demonstrated that 9 were immunocompetent (39%), 7 of whom had cutaneous infections (30%) with a predominant history of trauma (6/7), and 14 were immunosuppressed, 9 of whom were treated with prednisolone. Microbiology and histopathology were necessary in all cases for definite diagnosis. Among the 13 cases who underwent drug susceptibility testing, 10 cases were sensitive to trimethoprim-sulfamethoxazole (TMP-SMX) and 12 cases were sensitive to aminoglycoside. In conclusion, although N. otitidiscaviarum is one of the less commonly isolated species of Nocardia, it is capable of inducing localized or disseminated infection, even in an immunocompetent host. The majority of cases respond well to TMP-SMX and aminoglycoside, but the therapeutic action of cephalosporin is weak. Identification of bacteria and drug sensitivity tests for Nocardia is critical for guiding clinical treatment.

Specific clinical manifestations of Nocardia: A case report and literature review.

Nocardiosis is a rare bacterial infection of either the lungs (pulmonary) or body (systemic) that usually affects immunocompromised individuals. It is caused by Gram-positive, aerobic actinomycetes of the Nocardia genus. Multiple high-density sheet shadows in both lungs along with nodules or cavities are the most common presentations of nocardiosis, whereas a large pulmonary mass is considered to be rare. However, there is no specificity in the clinical manifestation of the disease. Therefore, isolation and identification of Nocardia strains is the only reliable diagnostic method. The present study describes the cases of two male patients of Asian descent with nocardiosis. Chest computed tomography scans showed a suspected tumor mass in both patients. Microscopic analysis and culturing of tissue samples obtained using a bronchoscope detected the presence of Nocardia wallacei. Neither patient showed signs of immunosuppression. The present study aimed to improve the understanding of lung nocardiosis and demonstrated that pulmonary nocardiosis should be suspected in the case of non-immunocompromised patients with a large mass in the lung. Furthermore, a review of the literature on infection with Nocardia was conducted.

Preclinical Studies of Mesenchymal Stem Cell (MSC) Administration in Chronic Obstructive Pulmonary Disease (COPD): A Systematic Review and Meta-Analysis.

BACKGROUND: In the last two decades, mesenchymal stem cells (MSCs) have been pre-clinically utilized in the treatment of a variety of kinds of diseases including chronic obstructive pulmonary disease (COPD). The aim of the current study was to systematically review and conduct a meta-analysis on the published pre-clinical studies of MSC administration in the treatment of COPD in animal models. METHODS AND RESULTS: A systematic search of electronic databases was performed. Statistical analysis was performed using the Comprehensive Meta-Analysis software (Version 3). The pooled Hedges's g with 95% confidence intervals (95% CIs) was adopted to assess the effect size. Random effect model was used due to the heterogeneity between the studies. A total of 20 eligible studies were included in the current systematic review. The overall meta-analysis showed that MSC administration was significantly in favor of attenuating acute lung injury (Hedges's g = -2.325 ± 0.145 with 95% CI: -2.609 ~ -2.040, P < 0.001 for mean linear intercept, MLI; Hedges's g = -3.488 ± 0.504 with 95% CI: -4.476 ~ -2.501, P < 0.001 for TUNEL staining), stimulating lung tissue repair (Hedges's g = 3.249 ± 0.586 with 95% CI: 2.103~ 4.394, P < 0.001) and improving lung function (Hedges's g = 2.053 ± 0.408 with 95% CI: 1.253 ~ 2.854, P< 0.001). The mechanism of MSC therapy in COPD is through ameliorating airway inflammation (Hedges's g = -2.956 ± 0.371 with 95% CI: -3.683 ~ -2.229, P< 0.001) and stimulating cytokine synthesis that involves lung tissue repair (Hedges's g = 3.103 ± 0.734 with 95% CI: 1.664 ~ 4.541, P< 0.001). CONCLUSION: This systematic review and meta-analysis suggest a promising role for MSCs in COPD treatment. Although the COPD models may not truly mimic COPD patients, these pre-clinical studies demonstrate that MSC hold promise in the treatment of chronic lung diseases including COPD. The mechanisms of MSCs role in preclinical COPD treatment may be associated with attenuating airway inflammation as well as stimulating lung tissue repair.

A Preliminary Study on RCN3 Protein Expression in Non-small Cell Lung Cancer.

BACKGROUND: Reticulocalbin 3 (RCN3), a member of CREC (Cab45/reticulocalbin/ ERC-45/calumenin) family protein, is located in the secretory pathway of endoplasmic reticulum (ER) of living cells. Disruption of RCN3 leads to failure of lung function in the mouse model. Although ER stress has been associated with the development of a variety of tumors, the role of RCN3 in development of non-small cell lung cancer (NSCLC) in human is unknown at present. METHODS: In this study a total of 41 paired NSCLC specimens (cancer group) and the adjacent normal tissues (control group) were obtained from patients undergoing lung lobectomy or pneumonectomy surgeries in Beijing Shijitan Hospital, Capital Medical University. The RCN3 mRNA and protein level in each clinical sample was determined using quantitative real time-PCR and immunoblotting, respectively. Immunohistochemistry analysis was utilized to compare the protein expressional patterns of RCN3 between the two clinical sample groups. RESULTS: Immunoblotting showed that levels of RCN3 protein in the NSCLC tissues were significantly lower than those in the control group (p < 0.001), suggesting ER stress is closely associated with the cancer cells. Accordingly, the ER stress protein GRP78 (glucose-regulated protein 78, also known as BIP) was remarkably upregulated in the cancer group (p < 0.05). Within the cancer group, a significant difference in RCN3 protein expression was observed in squamous cell carcinoma versus adenocarcinoma (p < 0.05). In the lung cancer group, however, RCN3 protein levels were not correlated with the age and the gender. In addition, RCN3 mRNA levels showed no significant difference between the cancer and the control groups, suggesting that the differential regulation of RCN3 is likely at post-transcription stage in NSCLC. CONCLUSIONS: Our study showed that RCN3 protein level was significantly down regulated in NSCLC, suggesting a potential correlation between RCN3 protein depletion and development of NSCLC. Although the exact cause-effect relationship between RCN3 and NSCLC needs to be further investigated, the study helps to shed additional lights on the molecular regulation of the lung cancer.

The Synergistic Mechanism of Pemetrexed Followed by Kanglaite was due to KLT Subsequently Inhibiting the Pemetrexed-Activated MAPK Signaling Pathway.

BACKGROUND: Kanglaite (KLT) can enhance the cytotoxic effects of chemotherapy; however, the underlying mechanism remains unclear. We investigated the mechanism underlying the cytotoxic synergy between KLT and pemetrexed in NSCLC cell lines. METHODS: A549 and H1975 cell lines were treated with pemetrexed and/or KLT in vitro. IC50 values, the combination index, cell cycle distribution, and signaling pathway analysis were assessed. RESULTS: Cytotoxic interactions between KLT and pemetrexed were dose-dependent in A549 and H1975 NSCLC cell lines. The administration of pemetrexed followed by KLT had a synergistic effect and an advantage over KLT followed by pemetrexed. Concomitant administration in both cell lines indicated that the cytotoxic interactions between KLT and pemetrexed were schedule-dependent. Cell cycle analysis showed that KLT arrested cells mainly in the G2/M phase, whereas pemetrexed arrested cells mainly in the S phase. Exposure to KLT first induced G2/M arrest and subsequently prevented the cytotoxicity of the S phase-specific drug pemetrexed. Signaling pathway analysis showed that exposure to pemetrexed resulted in increased phospho-p44/42MAPK levels which were inhibited by subsequent exposure to KLT. Thus pemetrexed followed by KLT inhibited the MAPK signaling pathway more obviously than KLT followed by pemetrexed. CONCLUSIONS: Pemetrexed followed by KLT had a synergistic effect and an advantage over other sequences in NSCLC cell lines. The synergistic mechanism was due to KLT subsequently inhibiting the pemetrexed-activated MAPK signaling pathway. These findings may provide molecular evidence to support clinical treatment strategies for patients with NSCLC.

Multicentre investigation of pathogenic bacteria and antibiotic resistance genes in Chinese patients with acute exacerbation of chronic obstructive pulmonary disease.

OBJECTIVE: A prospective observational study to investigate the distribution and antimicrobial resistance of pathogenic bacteria in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in Beijing, China. METHODS: Patients with AECOPD were recruited from 11 general hospitals. Sputum specimens were cultured and bacteria identified. Antibiotic susceptibility was determined for each isolate, and presence of antibiotic resistance genes was evaluated using polymerase chain reaction. RESULTS: Pathogenic bacteria were isolated from 109/318 patients (34.28%); 124 isolates of 22 pathogenic bacterial species were identified, including Klebsiella pneumoniae (16.94%), Pseudomonas aeruginosa (16.94%), Acinetobacter baumannii (11.29%), Streptococcus pneumoniae (8.87%), and Staphylococcus aureus (7.26%). S. aureus was sensitive to tigecycline, teicoplanin, vancomycin and linezolid but resistant to penicillin and levofloxacin. K.pneumoniae, P. aeruginosa, A. baumannii and E. coli were susceptible to amikacin and cefoperazone. CONCLUSIONS: K. pneumoniae and P. aeruginosa are the most common pathogenic bacteria in AECOPD cases in Beijing, China. Our antibiotic resistance findings may be helpful in selecting antibiotic therapy.

Attenuating effect of Ginsenoside Rb1 on LPS-induced lung injury in rats.

BACKGROUND: Sepsis causes neutrophil sequestration in the lung which leads to acute lung injury (ALI). Radix Ginseng (RG), a traditional herb used as herbal remedy in eastern Asia for thousands of years, which has been traditionally used in China to improve blood circulation and ameliorate pathological hemostasis. This study investigated whether Ginsenoside Rb1, the main components of RG, can attenuate ALI induced by LPS. METHODS: In vivo, 30 male Wistar rats were divided into three groups (n = 10 each groups) on the basis of the reagent used, which were subjected to LPS injection with or without Ginsenoside Rb1 (5 mg/kg) treatments to induce ALI model. Lung injury was assessed by pulmonary histology, lung wet-weight to dry-weight (W/D) ratio, the number of myeloperoxidase (MPO) positive cells, immunohistochemical analysis of intercellular adhesion molecule-1 (ICAM-1), gene expression of ICAM-1, ultrastructure changes of pulmonary microvasculature, concentration of inflammatory markers and in plasma. In vitro, pulmonary microvascular endothelial cells (PMVECs) were stimulated with LPS in the presence and absence of Ginsenoside Rb1 (50 mM), nuclear factor-κB (NF-κB) p65 was measured by immunocytochemistry staining and western blotting. RESULTS: Infusion of LPS induced lung injury, in vivo, as demonstrated by pulmonary edema with infiltration of neutrophils and hemorrhage, the increase in lung W/D ratio, the number of MPO positive cells, the level of inflammatory markers such as TNF-α, MCP-1 and IL-8, enhanced expression of ICAM-1 and ICAM-1 gene. Moreover, resulted in the changes of intercellular junctions in the endothelial cells of pulmonary microvasculature. In vitro, the significant increased release of NF-κB p65 and its subsequent translocation into the nucleus in PMVECs were observed. In contrast, Ginsenoside Rb1 treatment significantly ameliorated the LPS-induced lung injury, as judged by the marked improvement in all these indices. CONCLUSIONS: These results indicate that Ginsenoside Rb1 attenuated LPS-induced lung injury through an inhibition of the inflammatory signaling pathway, besides the direct inhibitory effect on proinflammatory molecules.

Analysis of growth factors in serum and induced sputum from patients with asthma.

Epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), the AA and BB isoforms of platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) are involved in the pathogenesis of airway inflammation in asthma. In the present study, the associations between asthmatic phenotypes and the expression levels of these mediators in induced sputum and serum were investigated. A total of 62 asthmatic patients were divided into eosinophilic or neutrophilic phenotypes by cytological classification of the induced sputum. In addition, patients were classified according to lung function (FEV1/FVC >70% or FEV1/FVC <70%) and asthma severity (mild, moderate or severe). The concentrations of EGF, bFGF, PDGF-AA, PDGF-BB and VEGF in the serum and induced sputum were measured using sandwich enzyme immunoassays. VEGF levels in the serum and induced sputum were higher in patients with an eosinophilic phenotype compared with those with a neutrophilic phenotype. In addition, VEGF expression was higher in patients with an FEV1/FVC value of <70% as compared with patients with an FEV1/FVC value of >70%. Furthermore, the levels of VEGF were higher in patients with severe asthma compared with the patients with mild and moderate asthma. There were no statistically significant differences observed with regard to EGF, bFGF, PDGF-AA and PDGF-BB levels among the various phenotypes. Therefore, the observations of the present study indicated that increased VEGF expression in the serum and induced sputum of patients may be associated with eosinophilic airway inflammation, severe airflow limitation and the severity of asthma.