Risk factors of in-hospital mortality in patients with pneumocystis pneumonia diagnosed by metagenomics next-generation sequencing.

Objectives: The metagenomic next-generation sequencing (mNGS) test is useful for rapid and accurate detection and identification of pathogenic microorganisms. The aim of the present study was to investigate the factors associated with in-hospital mortality in pneumocystis pneumonia (PCP) patients with mNGS-assisted diagnosis. Methods: Our study enrolled 154 patients with mNGS-positive PCP from August 2018 to February 2022 at the First Affiliated Hospital of Zhengzhou University respectively. Patients were divided into the survivor group (n=98) and the death group (n=56) according to whether in-hospital death occurred. Baseline characteristics, patients' pre-hospital symptoms and patients' CT imaging performance during hospitalization were carefully compared between the two groups. Risk factors for the occurrence of in-hospital death were sought by selecting indicators that were significantly different between the two groups for modelling and performing multiple logistic regression analysis. Results: Compared with the in-hospital death patients, the survivors were younger and had higher levels of albumin (ALB) (age: 50.29 ± 14.63 years vs 59.39 ± 12.27 years, p<0.001; ALB: 32.24 ± 5.62 g/L vs 29.34 ± 5.42g/L, p=0.002; respectively), while the levels of lactate dehydrogenase (LDH) and C-reactive protein CRP were lower (LDH: 574.67 ± 421.24 U/L vs 960.80 ± 714.94 U/L, p=0.001; CRP: 54.97 ± 55.92 mg/L vs80.45 ± 73.26 mg/L, p=0.018; respectively). Multiple logistic regression analysis revealed that age, the baseline LDH and CRP levels were all positively associated with high in-hospital mortality [age: OR(95%CI): 1.115 (1.062-1.172), p<0.001; LDH: OR(95%CI): 1.002 (1.001-1.003), p<0.001; CRP: OR(95%CI): 1.008 (1.000-1.017), p=0.045; respectively] while the platelet counts was negatively associated with it [OR(95%CI): 0.986 (0.979-0.992), p<0.001]. Conclusions: Old age, high baseline levels of LDH and CRP and low platelet counts were risk factors of the in-hospital mortality in mNGS positive PCP patients.

TGFß1-Smad Signaling Pathway Participates in Interleukin-33 Induced Epithelial-to-Mesenchymal Transition of A549 Cells.

BACKGROUNDS/AIMS: Epithelial-to-mesenchymal transition (EMT) has been proven to be involved in development and progression of pulmonary fibrosis. This study aims to investigate the role of transforming growth factor ß1 (TGFß1)-smad signaling pathway in the interleukin-33 (IL-33) induced EMT. METHODS: The human type II alveolar epithelial cell line, A549, and small airway epithelial cells (SAEC) were cultured and divided into 4 groups including Control, LY-2109761 (TGFß receptor inhibitor), IL-33 and IL-33+LY-2109761 group. Expression of TGFß1, E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) were examined by using real-time PCR (RT-PCR) and western blot assay, respectively. The smad3 signaling pathway factors, including smad3 and phosphorylated smad3 (p-smad3), were also detected by using western blot assay. RESULTS: IL-33 significantly activated T1/ST2 expression in A549 cells (P< 0.05). TGFß1 receptor inhibitor significantly suppressed the IL-33 caused down-expression of E-cad compared to IL-33 alone (P< 0.05). IL-33 significantly increased the α-SMA levels compared to Control group (P< 0.05) and TGFß1 receptor inhibitor inhibited the other effects of IL-33. IL-33 significantly enhanced the levels of TGFß1 compared to Control group (P< 0.05). TGFß1 receptor inhibitor suppressed the IL-33 induced up-expression of p-smad3. CONCLUSION: The TGFß1-smad signaling pathway participates in the IL-33 induced epithelial-to-mesenchymal transition of A549 cells.

Calpain 1 regulates TGF-ß1-induced epithelial-mesenchymal transition in human lung epithelial cells via PI3K/Akt signaling pathway.

Cell proliferation, transformation, and epithelial-mesenchymal transition (EMT) are key processes involved in the development of idiopathic pulmonary fibrosis (IPF). This study investigated the regulatory factors and signaling pathways that mediate EMT in the human type II alveolar epithelial A549 cell line. A549 cells were cultured in RPMI-1640 medium and allocated to the following four groups: blank control group or treated with transforming growth factor-ß1 (TGF-ß1), TGF-ß1 + PD 150606 (a calpain 1 inhibitor), or PD 150606. We examined E-cadherin (E-cad), α-smooth muscle actin (α-SMA), and calpain 1 mRNA transcript and protein expression levels in these four groups by performing RT-PCR and western blot analyses. The results indicated that TGF-ß1 treatment significantly downregulated E-cad and upregulated α-SMA expression compared with that of the blank control group (P<0.05). TGF-ß1 also enhanced calpain 1 expression compared with that of the blank control group (P<0.05). By contrast, treatment with the calpain 1 inhibitor PD 150606 increased E-cad expression and decreased α-SMA expression. Furthermore, PD 150606 treatment antagonized TGF-ß1-mediated increase in Akt/phospho-Akt in A549 epithelial cells. However, TGF-ß1-induced ETM was not correlated with the ERK and JNK signaling pathways. These combined results indicate that calpain 1 could regulate EMT in TGF-ß1-treated A549 epithelial cells via the PI3K/Akt signaling pathway.

Cigarette smoke induces the expression of Notch3, not Notch1, protein in lung adenocarcinoma.

The aim of the present study was to determine the effect of cigarette smoke on the expression of Notch proteins in lung adenocarcinoma (LAC). Protein expression levels of Notch1 and Notch3 were analyzed using immunohistochemistry in 102 human LAC specimens. Of these, 52 were obtained from smokers and 50 from non-smokers. In addition, cigarette smoke extract (CSE) at varying concentrations (1, 2.5 and 5%) was administered to A549 cells. The expression of Notch1 and Notch3 protein was then detected by western blot analysis at different time points (0, 8, 24 and 48 h). Of the 102 LAC specimens, 42 (41.2%) were positive for Notch1 and 63 (61.8%) were positive for Notch3. There was no significant difference in the level of Notch1 expression between smokers and non-smokers with LAC (P>0.05). The positive rate and staining intensity of Notch3 expression were increased in the smokers compared with the non-smokers (P<0.05). The expression of Notch3 protein in A549 cells increased in a time- and dose-dependent manner following treatment with CSE, whilst the expression of Notch1 protein appeared stable. The results suggested that cigarette smoke was able to induce the expression of Notch3, not Notch1, protein in LAC. The data revealed an upregulation of Notch3 in LAC following cigarette smoke exposure. Such findings may provide a novel therapeutic target for the treatment of LAC.

Beneficial Impact of bFGF Antisense Therapy in a Rat Model of Pulmonary Fibrosis.

OBJECTIVES: This study aims to explore basic fibroblast growth factor (bFGF)'s role in the development of pulmonary fibrosis through applying bFGF antisense oligonucleotide therapy in a rat model of pulmonary fibrosis. METHODS: Thirty rats were randomly divided into five groups: bFGF sense-transfected, bFGF antisense-transfected, null vector-transfected, pulmonary fibrosis (PF), and control groups. Sense, antisense, null, and PF groups were administered bleomycin to induce pulmonary fibrosis. Sense, antisense, and null vectors were intratracheally injected into the lungs of their respective groups followed by sacrifice after 28 days post-injection. Lung coefficients, H&E and Masson trichome staining, and serum and bronchoalveolar lavage fluid bFGF expression were comparatively assessed in addition to lung homogenate mRNA expressions of several select proteins and hydroxyproline content. RESULTS: The antisense and sense groups had significantly decreased and increased lung coefficients and pulmonary fibrosis than the PF and null groups, respectively, with the pulmonary fibrosis stage positively correlated with treatment. Antisense, PF, and null groups showed significantly reduced collagen fiber levels compared to the sense group. The antisense group displayed significantly lower serum and lavage fluid bFGF expression in addition to significantly lower bFGF, α-smooth muscle actin, Smad3, transforming growth factor-ß1, connective tissue growth factor, collagen I (and significantly higher Smad7) mRNA expression relative to the PF and null groups. The antisense and sense groups showed significantly higher hydroxyproline content relative to the PF and null groups. CONCLUSIONS: bFGF appears to promote collagen I synthesis and upregulates TGF-ß1/Smad signaling to promote lung fibroblast proliferation and differentiation in pulmonary fibrosis. bFGF antisense oligonucleotide therapy shows promise in preventing the development of pulmonary fibrosis, likely though a TGF-ß1/Smad-based signaling mechanism.

Lack of association between ACE insertion/deletion polymorphism and lung cancer: A meta-analysis.

OBJECTIVE: Previous case-control studies on the relation between the angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and lung cancer were controversial. In this study, we aimed to further evaluate the relation between the ACE gene I/D polymorphism and lung cancer. METHODS: We selected eight case-control studies related to the ACE gene I/D polymorphism and lung cancer by searching PubMed, EMBase, Chinese Biomedical Literature Database, and Wanfang database. We utilized Q-test and I (2) test to test the heterogeneity between each study. To merge the odds ratio (OR) and 95% confidence interval, we utilized the fixed-effects model during the analyses. RESULTS: The present study included 1612 patients with lung cancer and 1442 cancer-free control subjects. By meta-analysis, we did not find any association between the ACE gene I/D polymorphism and lung cancer in either genotype or allele distribution (DD vs. (ID+II): OR = 0.96, 95% CI (0.80-1.14), p = 0. 61; D allele vs. I allele: OR = 1.01, 95% CI (0.91-1.13), p = 0.79). CONCLUSION: We concluded that the ACE gene I/D polymorphism was not associated with lung cancer.