CSF3 aggravates acute exacerbation of pulmonary fibrosis by disrupting alveolar epithelial barrier integrity.

Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive respiratory disorder characterized by poor prognosis, often presenting with acute exacerbation. The primary cause of death associated with IPF is acute exacerbation of IPF (AE-IPF). However, the pathophysiology of acute exacerbation has not been clearly elucidated yet. This study aims to investigate the underlying pathophysiological molecular mechanism in a mouse AE-PF model. C57BL/6J mice were intratracheally administered bleomycin (BLM, 5 mg/kg) to induce pulmonary fibrosis. After 14 days, lipopolysaccharide (LPS, 2 mg/kg) was injected via the trachea route. Histological assessments, including H&E and Masson staining, as well as inflammatory indicators, were included to evaluate the induction of AE-PF by BLM and LPS in mice. Transcriptomic profiling of pulmonary tissues identified CSF3 as one of the top 10 upregulated DEGs in AE-PF mice. Indeed, administration of exogenous CSF3 protein exacerbated AE-PF in mice. Mechanistically, CSF3 disrupted alveolar epithelial barrier integrity and permeability by regulating specialized cell adhesion complexes such as tight junctions (TJs) and adherens junctions (AJs) via PI3K/p-Akt/Snail pathway, contributing to the aggravation of AE-PF in mice. Moreover, the discovery of elevated sera CSF3 indicated a notable increase in IPF patients during the exacerbation of the disease. Pearson correlation analysis in IPF patients revealed significant positive associations between CSF3 levels and KL-6 levels, LDH levels, CRP levels, respectively. These results provide mechanistic insights into the role of CSF3 in exacerbating of lung fibrotic disease and indicate monitoring CSF3 levels may aid in early clinical decisions for alternative therapy in the management of rapidly progressing IPF.

Does Market-Based Environmental Regulation Improve the Residents' Health: Quasi-Natural Experiment Based on DID.

Improving the residents' health is an important strategy for addressing the declining population dividend in China under the new development paradigm. Based on the panel data of 290 prefecture-level cities in China from 2010 to 2021, this paper uses environmental tax as a quasi-natural experiment, and adopts a DID model to explore the impact of market-based environmental regulation on the residents' health. The results show that the implementation of environmental tax can significantly reduce the population mortality rate, indicating an enhancement in residents' health outcomes. Mechanism analysis shows that environmental tax mainly relies on air quality to improve the residents' health. Also, the heath effect of environmental tax will be effective with the increase of income, and it's stronger in administrative border areas. Heterogeneity analysis shows that the effect of environmental tax on residents' health in western regions and resource-based cities is significantly stronger than those in central and eastern regions and non-resource-based cities. This paper provides new evidence for a comprehensive understanding of the impact of market-based environmental regulations on residents' well-being, offering insights for the implementation of green development strategies.

Facilitating or inhibiting? The impact of environmental information disclosure on enterprise investment value.

Environmental protection, which is beneficial for the present and the future, has become a global consensus, and environmental information disclosure (EID) is an effective way to realize and fulfill enterprise environmental responsibility. Although some scholars have studied the impact of EID on firms, there is less empirical evidence on the impact of EID on investors. In this study, we examine the impact of EID on enterprise investment value based on signaling theory using a time-varying difference-in-differences model and extract two channels of this effect. The study shows that the implementation of EID helps to enhance the value of enterprise investment. This enhancement will vary according to the location, the industry pollution type, and the nature of the enterprise: EID has a remarkable enhancement effect on the investment value of the eastern region, heavily polluted enterprises, and non-state-owned enterprises. To investigate the channel of EID's effect on enterprise investment value, we use the moderating effect model to analyze and find that enterprises with low tax ratios and small financing constraints can significantly enhance the effect of EID on investment value.

Have carbon emission trading pilot policy improved urban innovation capacity? Evidence from a quasi-natural experiment in China.

The carbon emission trading pilot policy is an important initiative to achieve synergistic economic-environmental development. Based on the data of 268 cities in China from 2006 to 2020, this paper analyzes the impact of carbon emission trading pilot policy on urban innovation capacity by using a time-varying difference-in-difference model. The study shows that, first, the implementation of the CETP improves the innovation capacity of cities, and the robustness test confirms the above findings. Second, the effect of the policy on enhancing urban innovation capacity is heterogeneous between the type of innovation and city type: the promotion of innovation capacity is stronger for utility model patents and non-capital cities. Third, there is a positive spillover effect of the implementation of the CETP on the promotion effect of the urban innovation capacity, which can lead to the improvement of the innovation capacity of neighboring cities. This paper has some reference value for building a unified carbon emission trading market and promoting low-carbon economic development within China.

Central vertical regulation and urban environment-biased technological progress: evidence from China.

Technological progress in favor of cleaner production is the key to achieving low-carbon development in China. The Ambient Air Quality Standard (AAQS) issued by the Ministry of Environmental Protection (MEP) in 2012 was an essential policy for the central government to implement vertical environmental regulation. Therefore, based on the city-level panel data, this paper examines the impact of the central vertical regulation on urban environment-biased technological progress using the difference-in-differences method. The results show that central vertical regulation can significantly promote urban environment-friendly technological progress. Heterogeneity analysis shows that the driving effect of the central vertical regulation on urban environment-friendly technological progress is more obvious in the eastern regions, non-resource-based cities, large cities, and high-grade cities. Moreover, the channel analysis shows that the central vertical regulation mainly boosts the urban environmental technology progress toward cleaner production by strengthening government environmental governance, raising public environmental concern, and improving energy structure. The findings provide policy implications for evaluating the effectiveness of macro-environmental policy and promoting green sustainable development.

Can environmental tax improve the environmental investment? Evidence from a quasi-natural experiment in China.

The implementation of the Environmental Tax Law is a milestone in promoting China's green tax reform. However, the existence literature has lacked attention to whether it leads enterprises to invest in green environmental protection. To examine the Environmental Tax Law effects and mechanism on the environmental investment of heavy-polluted enterprises, this study used the data of heavy-polluted enterprises listed on the A-share market from 2012 to 2020 and regarded the Environmental Tax Law as a quasi-natural experiment to employ a difference-in-differences model. We found that environmental tax improves the green environmental investment of heavy-polluted enterprises, reflecting the guiding role of policy on enterprise investment allocation. Heterogeneity was found, and the promotion effect of environmental tax reform on enterprise environmental investment is more significant in non-nation-owned, central-western regions, and small-scale enterprises. Further analysis believed that market competition, as an external mechanism, helps strengthen environmental tax reform's implementation effect. The findings of this paper provide a new proof for a comprehensive understanding of the micro-effect of environmental tax reform and provide a reference for the implementation of green development strategies.

Influence of public hospital reform on public health: Evidence from a quasi-natural experiment in China.

Public health is an important symbol of national wealth and prosperity. At present, China's public health is hindered by the poor management of public hospitals, which impacts the demographic structure and socioeconomic development. Therefore, taking the implementation of public hospital reform in China as a quasi-natural experiment, this study employed the time-varying DID model and the mediating effect to evaluate the influence of public hospital reform on public health. The results were as follows: (1) Public hospital reform can significantly improve public health, and a series of robustness tests have also confirmed the effects; (2) Government's financial support is a transmission mechanism for public hospital reform to promote public health; (3) After taking control variables into consideration, the effect of public hospital reform is stronger in the western region with a poorer economy. This research provides a vital policy reference for promoting the scope of reform and improving the health of the general public.

Can environmental tax reform promote carbon abatement of resource-based cities? Evidence from a quasi-natural experiment in China.

China is entering a new period characterized by reaching peak and carbon neutralization, and environmental taxes are increasingly crucial for breaking the "carbon curse" of resource-based cities. Accordingly, using the implementation of China's Environmental Protection Tax Law (EPT Law) as a quasi-natural experiment, this study utilizes the DID model to assess this environmental tax reform's effect in terms of reducing carbon emissions. The research results are as follows: (1) The environmental tax reform (ETR) reduced the intensity of carbon emissions; it additionally promoted reducing total carbon emissions from resource-based cities. (2) The carbon abatement effect can also be achieved by upgrading industrial structures and improving innovation in the area of green technology. (3) The ETR has impacted carbon abatement in resource-based cities more significantly in China's eastern region than in the central or western regions. In contrast, it had less effect on resource-based cities in the regenerative stage than on cities in other stages. (4) The spatial spillover effect of the ETR was significantly positive, aggravating the level of carbon emissions in neighboring cities. Thus, the "pollution haven hypothesis" was tested. Overall, this study deepens the knowledge of ETR and carbon emissions and provides theoretical support and policy suggestions for supporting resource-based cities in a green transformation.

Influence of carbon emission trading policy on residents' health in China.

Residents' health is the basic condition for economic and social development. At present, China's environmental pollution problem is becoming increasingly serious, which not only hinders sustainable economic and social development, but also poses a major threat to public health. Therefore, based on the carbon emissions trading policy implemented in China, this paper explores this policy's impact on residents' health using the DID model and illustrates the moderating effect of environmental pollution. The results show that (1) carbon emissions trading policies can promote the improvement of residents' health; (2) the effect is stronger for western regions and provinces with smaller population sizes after taking control variables into consideration; and (3) environmental pollution has a significant moderating effect on the relationship between carbon emissions trading and residents' health. This research serves as an important reference for expanding the scope of the policy pilot, reducing pollutant emissions, and improving the health of the population.

The effects of environmental tax reform on urban air pollution: A quasi-natural experiment based on the Environmental Protection Tax Law.

Air pollution significantly impacts sustainable development and public health. Taking the implementation of China's Environmental Protection Tax Law in China as a quasi-natural experiment, this paper employs the difference-in-differences (DID) and spatial DID models to evaluate the effects of environmental tax reform on urban air pollution. The findings are as follows. (1) Environmental tax reform can significantly reduce urban air pollution, and a series of robustness tests have also been conducted to provide further evidence. (2) Green technology innovation and industrial structure upgrading from a vital transmission mechanism for environmental tax reform to improve air quality. (3) Environmental tax reform significantly inhibits urban air pollution in cities located north of the Qinling-Huaihe line and big cities. (4) Moreover, environmental tax reform not only promotes the improvement of local air quality but also has a significant negative spatial spillover effect, reducing air pollution in neighboring cities. The research conclusions provide theoretical support and policy suggestions for promoting sustainable economic development, rationally optimizing environmental protection tax policies and improving urban air quality.

Resveratrol pretreatment mitigates LPS-induced acute lung injury by regulating conventional dendritic cells' maturation and function.

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a severe syndrome lacking efficient therapy and resulting in high morbidity and mortality. Although resveratrol (RES), a natural phytoalexin, has been reported to protect the ALI by suppressing the inflammatory response, the detailed mechanism of how RES affected the immune system is poorly studied. Pulmonary conventional dendritic cells (cDCs) are critically involved in the pathogenesis of inflammatory lung diseases including ALI. In this study, we aimed to investigate the protective role of RES via pulmonary cDCs in lipopolysaccharide (LPS)-induced ALI mice. Murine ALI model was established by intratracheally challenging with 5 mg/kg LPS. We found that RES pretreatment could mitigate LPS-induced ALI. Additionally, proinflammatory-skewed cytokines decreased whereas anti-inflammatory-related cytokines increased in bronchoalveolar lavage fluid by RES pretreatment. Mechanistically, RES regulated pulmonary cDCs' maturation and function, exhibiting lower level of CD80, CD86, major histocompatibility complex (MHC) II expression, and IL-10 secretion in ALI mice. Furthermore, RES modulated the balance between proinflammation and anti-inflammation of cDCs. Moreover, in vitro RES pretreatment regulated the maturation and function of bone marrow derived dendritic cells (BMDCs). Finally, the adoptive transfer of RES-pretreated BMDCs enhanced recovery of ALI. Thus, these data might further extend our understanding of a protective role of RES in regulating pulmonary cDCs against ALI.

Smad7 suppresses melanoma lung metastasis by impairing Tregs migration to the tumor microenvironment.

Transforming growth factor ß (TGF-ß) signaling plays critical roles in both physiological and pathological conditions. In the tumor microenvironment, TGF-ß are well demonstrated as a tumor inducer, which also promote tumor growth and metastasis. SMAD family is an important TGF-ß signalling transducer, which consists of receptor-regulated SMADs (R-SMADs), common-mediator SMADs (co-SMADs), and inhibitory SMADs (I-SMADs). Smad7 is one of the I-SMADs which has been proved to block TGF-ß signalling transduction in both tumor cells and immune cells. Accumulated evidence has suggested SMAD7 acted as a tumor suppressor in various cancer types, such as colorectal cancer, pancreatic cancer and skin melanoma, etc. However, the role of SMAD7 in melanoma lung metastasis has not been well studied. Here, we first investigated the role of SMAD7 on tumor cell viability by overexpressing SMAD7 in murine melanoma cell line B16-F10. Our results showed that SMAD7 overexpression slightly impaired B16-F10 cells growth, promoted cell apoptosis and arrested the cell cycle at S phase. In vivo study showed that SMAD7 overexpression inhibited B16-F10 lung metastasis. Further mechanism study suggested that SMAD7 promoted T cells activation by decreasing regulatory T cells (Tregs) infiltrating into the tumor microenvironment. In summary, our results proved that tumor cell derived SMAD7 inhibited melanoma lung metastasis by impairing the migration capacity of Tregs.

gB co-immunization with GP96 enhances pulmonary-resident CD8 T cells and exerts a long-term defence against MCMV pneumonitis.

Human cytomegalovirus (HCMV) infection in the respiratory tract leads to pneumonitis in immunocompromised hosts without available vaccine. Considering cytomegalovirus (CMV) mainly invades through the respiratory tract, CMV-specific pulmonary mucosal vaccine development that provides a long-lasting protection against CMV challenge gains our attention. In this study, N-terminal domain of GP96 (GP96-NT) was used as a mucosal adjuvant to enhance the induction of pulmonary-resident CD8 T cells elicited by MCMV glycoprotein B (gB) vaccine. Mice were intranasally co-immunized with 50 µg pgB and equal amount of pGP96-NT vaccine 4 times at 2-week intervals, and then i.n. challenged with MCMV at 16 weeks after the last immunization. Compared with pgB immunization alone, co-immunization with pgB/pGP96-NT enhanced a long-lasting protection against MCMV pneumonitis by significantly improved pneumonitis pathology, enhanced bodyweight, reduced viral burdens and increased survival rate. Moreover, the increased CD8 T cells were observed in lung but not spleen from pgB/pGP96-NT co-immunized mice. The increments of pulmonary CD8 T cells might be mainly due to non-circulating pulmonary-resident CD8 T-cell subset expansion but not circulating CD8 T-cell populations that home to inflammation site upon MCMV challenge. Finally, the deterioration of MCMV pneumonitis by depletion of pulmonary site-specific CD8 T cells in mice that were pgB/pGP96-NT co-immunization might be a clue to interpret the non-circulating pulmonary-resident CD8 T subset expansion. These data might uncover a promising long-lasting prophylactic vaccine strategy against MCMV-induced pneumonitis.

After Treatment with Methylene Blue is Effective against Delayed Encephalopathy after Acute Carbon Monoxide Poisoning.

Delayed encephalopathy after acute carbon monoxide (CO) poisoning (DEACMP) is the most severe and clinically intractable complication that occurs following acute CO poisoning. Unfortunately, the mechanism of DEACMP is still vague. Growing evidence indicates that delayed cerebral damage after CO poisoning is related to oxidative stress, abnormal neuro-inflammation, apoptosis and immune-mediated injury. Our recent report indicated that methylene blue (MB) may be a promising therapeutic agent in the prevention of neuronal cell death and cognitive deficits after transient global cerebral ischaemia (GCI). In this study, we aimed to investigate the potential of MB therapy to ameliorate the signs and symptoms of DEACMP. Rats were exposed to 1000 ppm CO for 40 min. in the first step; CO was then increased to 3000 ppm, which was maintained for another 20 min. The rats were implanted with 7-day release Alzet osmotic mini-pumps subcutaneously under the back skin, which provided MB at a dose of 0.5 mg/kg/day 1 hr after CO exposure. The results showed that MB significantly suppressed oxidative damage and expression of pro-inflammatory factors, including tumour necrosis factor-α and interleukin (IL)-1ß. MB treatment also suitably modulated mitochondrial fission and fusion, which is helpful in the preservation of mitochondrial function. Furthermore, MB dramatically attenuated apoptosis and neuronal death. Lastly, behavioural studies revealed that MB treatment preserved spatial learning and memory in the Barnes maze test. Our findings indicated that MB may have protective effects against DEACMP.

Involvement of IL-17 in Secondary Brain Injury After a Traumatic Brain Injury in Rats.

The pro-inflammatory activity of interleukin 17, which is produced by the IL-23/IL-17 axis, has been associated with the pathogenesis of traumatic brain injury (TBI). The study investigated the potential role of IL-17 in secondary brain injury of TBI in a rat model. Our data showed that the levels of IL-17 increased from 6 h to 7 days and peaked at 3 days, in both the CNS and serum, which were consistent with the severity of secondary brain injury. The IL-23 inhibitor suberoylanilide hydroxamic acid (SAHA) treatment markedly decreased the expressions of IL-17 and apoptosis-associated proteins cleaved caspase-3 and increased the protein ratio of Bcl-2 (B cell lymphoma/leukemia-2)/Bax (Bcl-2-associated X protein). Meanwhile, neuronal apoptosis was reduced, and neural function was improved after SAHA treatment. This study suggests that IL-17 is involved in secondary brain injury after TBI. Administering an IL-23 inhibitor and thereby blocking the IL-23/IL-17 axis may be beneficial in the treatment of TBI.

Self-oligomerization is essential for enhanced immunological activities of soluble recombinant calreticulin.

We have recently reported that calreticulin (CRT), a luminal resident protein, can be found in the sera of patients with rheumatoid arthritis and also that recombinant CRT (rCRT) exhibits extraordinarily strong immunological activities. We herein further demonstrate that rCRT fragments 18-412 (rCRT/18-412), rCRT/39-272, rCRT/120-308 and rCRT/120-250 can self-oligomerize in solution and are 50-100 fold more potent than native CRT (nCRT, isolated from mouse livers) in activating macrophages in vitro. We narrowed down the active site of CRT to residues 150-230, the activity of which also depends on dimerization. By contrast, rCRT/18-197 is almost completely inactive. When rCRT/18-412 is fractionated into oligomers and monomers by gel filtration, the oligomers maintain most of their immunological activities in terms of activating macrophages in vitro and inducing specific antibodies in vivo, while the monomers were much less active by comparison. Additionally, rCRT/18-412 oligomers are much better than monomers in binding to, and uptake by, macrophages. Inhibition of macrophage endocytosis partially blocks the stimulatory effect of rCRT/18-412. We conclude that the immunologically active site of CRT maps between residues 198-230 and that soluble CRT could acquire potent immuno-pathological activities in microenvironments favoring its oligomerization.

A monoclonal antibody specific to human myxovirus resistance protein A.

Recombinant human myxovirus resistance protein A (MxA) was successfully expressed by an Escherichia coli expression system. After immunization and cell fusion, a mouse hybridoma (3C2) producing MAbs to MxA was established. Hybridoma 3C2 was further characterized using indirect ELISA, Western blot analysis, immunofluorescent staining, and immunoprecipitation. The ELISA results showed that the titer of 3C2 was between 1:6400 and 1:12800 in ascitic fluids. The isotype of the monoclonal antibody was tested to be IgG1kappa. 3C2 can also specifically recognize human MxA protein in various formats by Western blot analysis, immunofluorescent staining, and immunoprecipitation assay. We further demonstrated that 3C2 could be used to detected MxA expression induce by type I interferon in A549 cell line and human peripheral blood mononuclear cells by Western blot in a dose-dependent manner.

Shell-sheddable micelles based on dextran-SS-poly(epsilon-caprolactone) diblock copolymer for efficient intracellular release of doxorubicin.

Reduction-responsive biodegradable micelles were developed from disulfide-linked dextran-b-poly(epsilon-caprolactone) diblock copolymer (Dex-SS-PCL) and applied for triggered release of doxorubicin (DOX) in vitro and inside cells. Dex-SS-PCL was readily synthesized by thiol-disulfide exchange reaction between dextran orthopyridyl disulfide (Dex-SS-py, 6000 Da) and mercapto PCL (PCL-SH, 3100 Da). Dynamic light scattering (DLS) measurements showed that Dex-SS-PCL yielded micelles with an average size of about 60 nm and a low polydispersity index (PDI 0.1-0.2) in PB (50 mM, pH 7.4). Interestingly, these micelles formed large aggregates rapidly in response to 10 mM dithiothreitol (DTT), most likely due to shedding of the dextran shells through reductive cleavage of the intermediate disulfide bonds. DOX could be efficiently loaded into the micelles with a drug loading efficiency of about 70%. Notably, the in vitro release studies revealed that Dex-SS-PCL micelles released DOX quantitatively in 10 h under a reductive environment, mimicking that of the intracellular compartments such as cytosol and the cell nucleus, whereas only about 27% DOX was released from reduction insensitive Dex-PCL micelles in 11.5 h under otherwise the same conditions and about 20% DOX released from Dex-SS-PCL micelles in 20 h under the nonreductive conditions. The cell experiments using fluorescence microscopy and confocal laser scanning microscopy (CLSM) showed clearly that DOX was rapidly released to the cytoplasm as well as to the cell nucleus. MTT studies revealed a markedly enhanced drug efficacy of DOX-loaded Dex-SS-PCL micelles as compared to DOX-loaded reduction-insensitive Dex-PCL micelles. These reduction-responsive biodegradable micelles have appeared highly promising for the targeted intracellular delivery of hydrophobic chemotherapeutics in cancer therapy.