GPS2 ameliorates cigarette smoking-induced pulmonary vascular remodeling by modulating the ras-Raf-ERK axis.

BACKGROUND: Mitogen-activated protein kinase (MAPK)signaling-mediated smoking-associated pulmonary vascular remodeling (PVR) plays an important role in the pathogenesis of group 3 pulmonary hypertension (PH). And G protein pathway suppressor 2 (GPS2) could suppress G-protein signaling such as Ras and MAPK, but its role in cigarette smoking -induced PVR (CS-PVR) is unclear. METHODS: An in vivo model of smoke-exposed rats was constructed to assess the role of GPS2 in smoking-induced PH and PVR. In vitro, the effects of GPS2 overexpression and silencing on the function of human pulmonary arterial smooth cells (HPASMCs) and the underlying mechanisms were explored. RESULTS: GPS2 expression was downregulated in rat pulmonary arteries (PAs) and HPASMCs after CS exposure. More importantly, CS-exposed rats with GPS2 overexpression had lower right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), and wall thickness (WT%) than those without. And enhanced proliferation and migration of HPASMCs induced by cigarette smoking extract (CSE) can be evidently inhibited by overexpressed GPS2. Besides, GPS2siRNA significantly enhanced the proliferation, and migration of HPASMCs as well as activated Ras and Raf/ERK signaling, while these effects were inhibited by zoledronic acid (ZOL). In addition, GPS2 promoter methylation level in rat PAs and HPASMCs was increased after CS exposure, and 5-aza-2-deoxycytidine (5-aza) inhibited CSE-induced GPS2 hypermethylation and downregulation in vitro. CONCLUSIONS: GPS2 overexpression could improve the CS-PVR, suggesting that GPS2 might serve as a novel therapeutic target for PH-COPD in the future.

Optimal Decoding Order and Power Allocation for Sum Throughput Maximization in Downlink NOMA Systems.

In this paper, we consider a downlink non-orthogonal multiple access (NOMA) system over Nakagami-m channels. The single-antenna base station serves two single-antenna NOMA users based on statistical channel state information (CSI). We derive the closed-form expression of the exact outage probability under a given decoding order, and we also deduce the asymptotic outage probability and diversity order in a high-SNR regime. Then, we analyze all the possible power allocation ranges and theoretically prove the optimal power allocation range under the corresponding decoding order. The demarcation points of the optimal power allocation ranges are affected by target data rates and total power, without an effect from the CSI. In particular, the values of the demarcation points are proportional to the total power. Furthermore, we formulate a joint decoding order and power allocation optimization problem to maximize the sum throughput, which is solved by efficiently searching in our obtained optimal power allocation ranges. Finally, Monte Carlo simulations are conducted to confirm the accuracy of our derived exact outage probability. Numerical results show the accuracy of our deduced demarcation points of the optimal power allocation ranges. And the optimal decoding order is not constant at different total transmit power levels.

CXCL17 Attenuates Diesel Exhaust Emissions Exposure-Induced Lung Damage by Regulating Macrophage Function.

Exposure to diesel exhaust emissions (DEE) is strongly linked to innate immune injury and lung injury, but the role of macrophage chemoattractant CXCL17 in the lung damage caused by DEE exposure remains unclear. In this study, whole-body plethysmography (WBP), inflammatory cell differential count, and histopathological analysis were performed to assess respiratory parameters, airway inflammation, and airway injury in C57BL/6 male mice exposed to DEE for 3 months. qRT-PCR, IHC (immunohistochemistry), and ELISA were performed to measure the CXCL17 expression in airway epithelium or BALF (bronchoalveolar lavage fluid) following DEE/Diesel exhaust particle (DEP) exposure. Respiratory parameters, airway inflammation, and airway injury were assessed in CXCL17-overexpressing mice through adeno-associated virus vector Type 5 (AAV5) infection. Additionally, an in vitro THP-1 and HBE co-culture system was constructed. Transwell assay was carried out to evaluate the effect of rh-CXCL17 (recombinant human protein-CXCL17) on THP-1 cell migration. Flow cytometry and qRT-PCR were conducted to assess the impacts of rh-CXCL17 on apoptosis and inflammation/remodeling of HBE cells. We found that the mice exposed to DEE showed abnormal respiratory parameters, accompanied by airway injury and remodeling (ciliary injury in airway epithelium, airway smooth muscle hyperplasia, and increased collagen deposition). Carbon content in airway macrophages (CCAM), but not the number of macrophages in BALF, increased significantly. CXCL17 expression significantly decreased in mice airways and HBE after DEE/DEP exposure. AAV5-CXCL17 enhanced macrophage recruitment and clearance of DEE in the lungs of mice, and it improved respiratory parameters, airway injury, and airway remodeling. In the THP-1/HBE co-culture system, rh-CXCL17 increased THP-1 cell migration while attenuating HBE cell apoptosis and inflammation/remodeling. Therefore, CXCL17 might attenuate DEE-induced lung damage by recruiting and activating pulmonary macrophages, which is expected to be a novel therapeutic target for DEE-associated lung diseases.

HEG1 as a novel potential biomarker for the prognosis of lung adenocarcinoma.

BACKGROUND: Heart development protein with EGF-like domains 1 (HEG1), generally related to angiogenesis and embryonic development, was reported to participate in the occurrence and progression of some tumors recently. However, the role of HEG1 in lung adenocarcinoma (LUAD) is unclear. PATIENTS AND METHODS: To explore the effect of HEG1 on LUAD, GEPIA platform and UALCAN database, as well as Kaplan-Meier plotter were adopted to analyze the association of HEG1 with clinicopathological characteristics and survival outcomes for LUAD firstly. And then the HEG1 in LUAD tissues, blood and cell lines were detected by qRT-PCR, western blot, immunofluorescence, immunohistochemistry, and ELISA. Gene set enrichment analysis (GSEA) was conducted to identify pathways that might be affected by HEG1 in LUAD. RESULTS: In this study, HEG1 in lung tissues and cell lines of LUAD were significantly downregulated compared to benign pulmonary disease tissues and alveolar epithelial cells (p < 0.05). Moreover, compared with other groups, patients with advanced tumor stage had lower HEG1 mRNA expression levels (p = 0.025), which were negatively correlated with Ki67 index in tumor tissues (r = -0.427, p = 0.033). On the other hand, the LUAD patients with lower HEG1 had shorter overall survival (OS) (HR = 0.51, 95% CI: 0.40-0.65, p < 0.001) according to Kaplan-Meier plotter. In addition, HEG1 in serum of LUAD patients was negatively associated with CEA (r = -0.636, p < 0.001). GSEA showed that HEG1 was enriched in various metabolic-related pathways, including glucose metabolism, lipid metabolism, and nucleotide metabolism signaling. CONCLUSIONS: HEG1 was downregulated in LUAD patients and associated with poor prognosis, which indicating HEG1 may serve as a potential biomarker for diagnosis and prognosis of LUAD.

A Simple Clinical Prediction Tool for COVID-19 in Primary Care with Epidemiology: Temperature-Leukocytes-CT Results.

BACKGROUND Effective identification of patients with suspected COVID-19 is vital for the management. This study aimed to establish a simple clinical prediction model for COVID-19 in primary care. MATERIAL AND METHODS We consecutively enrolled 60 confirmed cases and 152 suspected cases with COVID-19 into the study. The training cohort consisted of 30 confirmed and 78 suspected cases, whereas the validation cohort consisted of 30 confirmed and 74 suspected cases. Four clinical variables - epidemiological history (E), body temperature (T), leukocytes count (L), and chest computed tomography (C) - were collected to construct a preliminary prediction model (model A). By integerizing coefficients of model A, a clinical prediction model (model B) was constructed. Finally, the scores of each variable in model B were summed up to build the ETLC score. RESULTS The preliminary prediction model A was Logit (YA)=2.657X1+1.153X2+2.125X3+2.828X4-10.771, while the model B was Logit (YB)=2.5X1+1X2+2X3+3X4-10. No significant difference was found between the area under the curve (AUC) of model A (0.920, 95% CI: 0.875-0.953) and model B (0.919, 95% CI: 0.874-0.952) (Z=0.035, P=0.972). When ETLC score was more than or equal to 9.5, the sensitivity and specificity for COVID-19 was 76.7% (46/60) and 90.1% (137/152), respectively, and the positive and negative predictive values were 75.4% (46/61) and 90.7% (137/151), respectively. CONCLUSIONS The ETLC score is helpful for efficiently identifying patients with suspected COVID-19.

CC16-TNF-α negative feedback loop formed between Clara cells and normal airway epithelial cells protects against diesel exhaust particles exposure-induced inflammation.

CC16 is almost exclusively expressed in non-ciliated epithelial Clara cells, and widely used as a Clara cell marker. Diesel exhaust particles (DEPs), the fine particulate matters produced by diesel engines, cause or exacerbate airway-related diseases. Our previous study documented that DEP inhibits the CC16 expression in the immortalized mouse Clara cell line through methylation of C/EBPα promoter. However, the molecular mechanism by which DEP regulates CC16 secretion is unclear. Here, we isolated CC16 containing Clara cells (CC16+) from human distal lung, and found that DEP inhibited CC16 secretion from CC16+ cells via methylation of C/EBPα and inhibition of Munc18b transcription. CC16+ cell conditioned media containing different concentrations of CC16 was prepared and used for culture of airway epithelial cells BEAS-2B with no expression of CC16. A positive correlation was observed between CC16 level and DEP-induced autophagy activity, and a negative correlation between CC16 level and DEP-induced pro-inflammatory cytokine TNF-α, IL-6, and IL-8 level, suggesting that CC16 might mitigate DEP-induced inflammation via promoting autophagy in BEAS-2B cells. This result was further confirmed by adding recombinant CC16 to BEAS-2B cells exposed to DEP. Moreover, CC16 level was significantly increased when CC16+ cells were cultured in BEAS-2B cell conditioned medium containing TNF-α or the normal medium supplemented with recombinant TNF-α, suggesting that TNF-α induced CC16 production and secretion from CC16+ cells. Collectively, these data point that CC16 and TNF-α form a negative feedback loop, and this negative feedback loop between Clara cells and normal airway epithelial cells protects against DEP exposure-induced inflammation.

Panaxydol attenuates ferroptosis against LPS-induced acute lung injury in mice by Keap1-Nrf2/HO-1 pathway.

BACKGROUND: Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) induces uncontrolled and self-amplified pulmonary inflammation, and has high morbidity and mortality rates in critically ill patients. In recent years, many bioactive ingredients extracted from herbs have been reported to effectively ameliorate ALI/ARDS via different mechanisms. Ferroptosis, categorized as regulated necrosis, is more immunogenic than apoptosis and contributes to the progression of ALI. In this study, we examined the impact of panaxydol (PX), isolated from the roots of Panax ginseng, on lipopolysaccharide (LPS)-induced ALI in mice. METHODS: In vivo, the role of PX on LPS-induced ALI in mice was tested by determination of LPS-induced pulmonary inflammation, pulmonary edema and ferroptosis. In vitro, BEAS-2B cells were used to investigate the molecular mechanisms by which PX functions via determination of inflammation, ferroptosis and their relationship. RESULTS: Administration of PX protected mice against LPS-induced ALI, including significantly ameliorated lung pathological changes, and decreased the extent of lung edema, inflammation, and ferroptosis. In vitro, PX inhibited LPS-induced ferroptosis and inflammation in bronchial epithelial cell line BEAS-2B cells. The relationship between ferroptosis and inflammation was investigated. The results showed that ferroptosis mediated inflammation in LPS-treated BEAS-2B cells, and PX might ameliorate LPS-induced inflammation via inhibiting ferroptosis. Meanwhile, PX could upregulate Keap1-Nrf2/HO-1 pathway, and selective inhibition of Keap1-Nrf2/HO-1 pathway significantly abolished the anti-ferroptotic and anti-inflammatory functions of PX in LPS-treated cells. CONCLUSION: PX attenuates ferroptosis against LPS-induced ALI via Keap1-Nrf2/HO-1 pathway, and is a promising novel therapeutic candidate for ALI.

Upregulation of LncRNA Malat1 Induced Proliferation and Migration of Airway Smooth Muscle Cells via miR-150-eIF4E/Akt Signaling.

The increased proliferation and migration of airway smooth muscle cells (ASMCs) are critical processes in the formation of airway remodeling in asthma. Long non-coding RNAs (lncRNAs) have emerged as key mediators of diverse physiological and pathological processes, and are involved in the pathogenesis of various diseases, including asthma. LncRNA Malat1 has been widely reported to regulate the proliferation and migration of multiple cell types and be involved in the pathogenesis of various human diseases. However, it remains unknown whether Malat1 regulates ASMC proliferation and migration. Here, we explored the function of Malat1 in ASMC proliferation and migration in vitro stimulated by platelet-derived growth factor BB (PDGF-BB), and the underlying molecular mechanism involved. The results showed that Malat1 was significantly upregulated in ASMCs treated with PDGF-BB, and knockdown of Malat1 effectively inhibited ASMC proliferation and migration induced by PDGF-BB. Our data also showed that miR-150 was a target of Malat1 in ASMCs, and inhibited PDGF-BB-induced ASMC proliferation and migration, whereas the inhibition effect was effectively reversed by Malat1 overexpression. Additionally, translation initiation factor 4E (eIF4E), an important regulator of Akt signaling, was identified to be a target of miR-150, and both eIF4E knockdown and Akt inhibitor GSK690693 inhibited PDGF-BB-induced ASMC proliferation and migration. Collectively, these data indicate that Malat1, as a competing endogenous RNA (ceRNA) for miR-150, derepresses eIF4E expression and activates Akt signaling, thereby being involved in PDGF-BB-induced ASMC proliferation and migration. These findings suggest that Malat1 knockdown may present a new target to limit airway remodeling in asthma.

Role of C/EBPα hypermethylation in diesel engine exhaust exposure-induced lung inflammation.

Exposure to diesel engine exhaust (DEE) impairs lung function. But the underlying mechanisms are still not fully understood. The aim of this study was to investigate the effects of long-term DEE exposure on lung inflammation and the underlying mechanisms. Sprague-Dawley male rats were exposed to DEE with 3 mg/m3 of diesel exhaust particles (DEP) for 12 weeks. Then urine, blood, bronchoalveolar lavage fluid (BALF), and lung tissue were collected for the determination of biochemistry indexes, DNA methylation status, and histological changes in the lung. The results showed that the metabolites of polycyclic aromatic hydrocarbons (PAHs) 2-hydroxyphenanthrene (2-OHPh) and 9-OHPh, and 8-hydroxy-2'-deoxyguanosine (8-OHdG), and malondialdehyde (MDA) level were higher in urine of DEE-exposed rats than control group. The level of proinflammatory cytokines IL-8, IL-6, and TNF-α was significantly higher in serum (1.8, 3.5, and nearly 1.0-fold increase, respectively), BALF (2.2, 3.8, and 2.0-fold increase, respectively) and lung tissues (3.5, 4.3, and 2.4-fold increase, respectively) of DEE-exposed rats than control group. While the level of clara cell secretory protein (CC16) and pulmonary surfactant protein D (SP-D) with anti-inflammatory property was obviously lower in serum (reduction of 29% and 38%, respectively), BALF (reduction of 50% and 46%, respectively) and lung tissues (reduction of 50% and 55%, respectively) of DEE-exposed rats than control group. Exposure to DEE also resulted in significant increases in total white blood cell (WBC), neutrophil, eosinophil, and lymphocyte number in BALF. Airway inflammation and remolding were apparent in DEE group. The methylation level of CCAAT/enhancer-binding protein alpha (C/EBPα) promoter was markedly increased (about 3.2-fold increase), and its mRNA and protein expression were significantly decreased (about 62% and 68% decrease, respectively) in the lungs of DEE-exposed rats compared with the group. Further, cell experiments were performed to investigate the relationship between C/EBPα and CC16, and CC16 function under DEP conditions. The results showed that DEP inhibited CC16 expression via methylation of C/EBPα promoter, and the increase of CC16 level significantly relieved the proinflammatory effects caused by DEP exposure. In conclusion, our data indicated that long-term exposure to DEE can cause lung inflammation, at least in part via methylation of C/EBPα promoter, and inhibition of CC16 expression.

Leptin positively regulates MUC5AC production and secretion induced by interleukin-13 in human bronchial epithelial cells.

Mucus hypersecretion and plugging of lower respiratory tract airways due to mucus plugs have long been recognized as the leading cause of the morbidity and mortality in asthma. MUC5AC protein is a major component of airway mucus. Here, we showed that interleukin (IL)-13 induced MUC5AC production and secretion, and leptin expression in the human bronchial epithelial cell line-16 (HBE16) cells in a concentration-dependent manner. Leptin knockdown suppressed MUC5AC production and secretion induced by IL-13. We further investigated the molecular mechanism by which leptin functioned, and found that leptin regulated IL-13-induced MUC5AC production and secretion via the JAK2-STAT3 pathway. Subsequently, Munc18b, a limiting component of the exocytic machinery of airway epithelial and mast cells, was found that when knockdown, MUC5AC secretion was significantly inhibited. SABiosciences ChIP search tool identified three STAT3 binding sites with Munc18b promoter. Chromatin immunoprecipitation analysis further confirmed that Stat3 upregulated Munc18b expression by directly binding to its promoter. These data suggested that leptin promotes MUC5AC secretion via JAK2-STAT3-MUNC18b regulatory network. Taken together, our data highlight a positive feedback role and molecular mechanism for leptin in the control of MUC5AC production and secretion from airway epithelial cells stimulated by IL-13, which encourage further exploration of the therapeutic potentials of manipulating leptin in the treatment of mucus hypersecretion in chronic inflammation lung diseases.

[Outcome and relevant factors of tubal pregnancy treated with laparoscopic conservative surgery].

OBJECTIVE: To investigate the therapeutic outcome and its influencing factors after laparoscopic conservative surgery in treatment of tubal pregnancy. METHODS: From January 2003 to December 2008, 226 cases with tubal pregnancy were treated by laparoscopic conservative surgery. The tubal patency was evaluated in 152 cases given by hysterosalpingography (HSG) and 6 cases given by second laparoscopic exploration at 3 - 6 months after surgery. In their first laparoscopic surgeries, 209 got successful treatment and 19 underwent fail treatment. At 3 - 6 months after surgery, 89 cases with tubal patency among 207 cases with successful treatment were enrolled in group A. Nineteen cases who were failed in their first laparoscopic conservative surgery and treated by salpingectomy and 63 cases with tubal obstruction were enrolled in group B. The rate of tubal patency was calculated on patients with characteristics of gestational sac less or more than 5 cm, the level serum human chorionic gonadotropin (hCG) less than 2000 IU/L, 2000 IU/L to 5000 IU/L, and more than 5000 IU/L. RESULTS: There was no significant difference in age, parity, amenorrhea, location of tubal pregnancy, rupture, pelvic adhesion between group A and group B. Two hundred and seven cases (91.6%, 207/226) were successfully treated at initial laparoscopy. One hundred and fifty-two cases got follow up and 55 cases lost follow up at 3 to 6 months after surgery. There was statistical difference in preoperative hCG value which median were 980 (55 - 12 000) IU/L in group A, 3150 (570 - 40 000) IU/L in group B (P < 0.01); the diameter of tubal gestational sac were (3.4 +/- 1.3) cm in group A and (5.0 +/- 1.7) cm in group B (P < 0.01); respectively, the volume of peritoneal bleeding were 200 (0 - 1500) ml and 300 (0 - 1600) ml, the rate of live tubal embryo was 2% (2/89) in group A and 11% (9/82) in group B, which all reached statistical difference (P < 0.05). Among 171 cases in both group A and B, the rate of tubal patency were 65% (67/103) in 103 cases with maximal diameter of tubal gestational sac less than 5 cm and 32% (22/68) in 68 cases with maximal diameter of tubal gestational sac more than 5 cm, which reached statistical difference (P < 0.01). The rate were 72% (73/102) in patients with serum level of hCG less than 2000 IU/L, 29% (12/42) in patients with 2000 IU/L to 5000 IU/L and 15% (4/27) in patients with more than 5000 IU/L, which also showed statistical difference (P < 0.05). It was observed that preoperative serum hCG level (OR = 0.277, P < 0.01), the maximal diameter of gestational sac (OR = 0.577, P < 0.01) and the volume of peritoneal bleeding (OR = 0.999, P < 0.05) were significant factors influencing successful laparoscopy treatment by logistical regression analysis. CONCLUSION: In order to preserve fertility, laparoscopic conservative surgery was a safe and feasible approach in treatment of tubal pregnancy. Preoperative serum hCG levels, size of tube gestational sac were significant factors influencing successful laparoscopic surgery.