Treatment Strategies for Anti-VEGF Resistance in Neovascular Age-Related Macular Degeneration by Targeting Arteriolar Choroidal Neovascularization.

Despite extensive use of intravitreal anti-vascular endothelial growth factor (anti-VEGF) biologics for over a decade, neovascular age-related macular degeneration (nAMD) or choroidal neovascularization (CNV) continues to be a major cause of irreversible vision loss in developed countries. Many nAMD patients demonstrate persistent disease activity or experience declining responses over time despite anti-VEGF treatment. The underlying mechanisms of anti-VEGF resistance are poorly understood, and no effective treatment strategies are available to date. Here we review evidence from animal models and clinical studies that supports the roles of neovascular remodeling and arteriolar CNV formation in anti-VEGF resistance. Cholesterol dysregulation, inflammation, and ensuing macrophage activation are critically involved in arteriolar CNV formation and anti-VEGF resistance. Combination therapy by neutralizing VEGF and enhancing cholesterol removal from macrophages is a promising strategy to combat anti-VEGF resistance in CNV.

Characterization of Vascular Morphology of Neovascular Age-Related Macular Degeneration by Indocyanine Green Angiography.

Age-related macular degeneration (AMD) is a leading cause of blindness among older individuals, and its prevalence is rapidly increasing due to the aging population. Choroidal neovascularization (CNV) or wet AMD, which accounts for 10%-20% of all AMD cases, is responsible for an alarming 80%-90% of AMD-related blindness. Current anti-VEGF therapies show suboptimal responses in approximately 50% of patients. Resistance to anti-VEGF treatment in CNV patients is often associated with arteriolar CNV, while responders tend to have capillary CNV. While fluorescein angiography (FA) is commonly used to assess leakage patterns in wet AMD patients and animal models, it does not provide information about CNV vascular morphology (arteriolar CNV vs. capillary CNV). This protocol introduces the use of indocyanine green angiography (ICGA) to characterize lesion types in laser-induced CNV mouse models. This method is crucial for investigating the mechanisms and treatment strategies for anti-VEGF resistance in wet AMD. It is recommended to incorporate ICGA alongside FA for comprehensive assessment of both leakage and vascular features of CNV in mechanistic and therapeutic studies.

Adverse short-term effects of ozone on cardiovascular mortalities modified by season and temperature: a time-series study.

Introduction: Ambient ozone pollution becomes critical in China. Conclusions on the short-term effects of ozone on cardiovascular mortality have been controversial and limited on cause-specific cardiovascular mortalities and their interactions with season and temperature. This research aimed to investigate the short-term effects of ozone and the modifications of season and temperature on cardiovascular mortality. Methods: Cardiovascular death records, air pollutants, and meteorological factors in Shenzhen from 2013 to 2019 were analyzed. Daily 1-h maximum of ozone and daily maximum 8-h moving average of ozone were studied. Generalized additive models (GAMs) were applied to evaluate their associations with cardiovascular mortalities in sex and age groups. Effect modifications were assessed by stratifying season and temperature. Results: Distributed lag impacts of ozone on total cardiovascular deaths and cumulative effects on mortality due to ischemic heart disease (IHD) were most significant. Population under 65 years old was most susceptible. Majority of significant effects were found in warm season, at high temperature, and at extreme heat. Ozone-associated risks in total deaths caused by hypertensive diseases reduced in warm season, while risks in IHD in males increased at high temperature. Extreme heat enhanced ozone effects on deaths caused by CVDs and IHD in the population under 65 years old. Discussion: The revealed cardiovascular impacts of ozone below current national standard of air quality suggested improved standards and interventions in China. Higher temperature, particularly extreme heat, rather than warm season, could significantly enhance the adverse effects of ozone on cardiovascular mortality in population under 65 years old.

Exploring the contribution of ARMS2 and HTRA1 genetic risk factors in age-related macular degeneration.

Age-related macular degeneration (AMD) is the leading cause of severe irreversible central vision loss in individuals over 65 years old. Genome-wide association studies (GWASs) have shown that the region at chromosome 10q26, where the age-related maculopathy susceptibility (ARMS2/LOC387715) and HtrA serine peptidase 1 (HTRA1) genes are located, represents one of the strongest associated loci for AMD. However, the underlying biological mechanism of this genetic association has remained elusive. In this article, we extensively review the literature by us and others regarding the ARMS2/HTRA1 risk alleles and their functional significance. We also review the literature regarding the presumed function of the ARMS2 protein and the molecular processes of the HTRA1 protein in AMD pathogenesis in vitro and in vivo, including those of transgenic mice overexpressing HtrA1/HTRA1 which developed Bruch's membrane (BM) damage, choroidal neovascularization (CNV), and polypoidal choroidal vasculopathy (PCV), similar to human AMD patients. The elucidation of the molecular mechanisms of the ARMS2 and HTRA1 susceptibility loci has begun to untangle the complex biological pathways underlying AMD pathophysiology, pointing to new testable paradigms for treatment.

Associations Among Multimorbid Conditions in Hospitalized Middle-aged and Older Adults in China: Statistical Analysis of Medical Records.

BACKGROUND: Multimorbidity has become a new challenge for medical systems and public health policy. Understanding the patterns of and associations among multimorbid conditions should be given priority. It may assist with the early detection of multimorbidity and thus improve quality of life in older adults. OBJECTIVE: This study aims to comprehensively analyze and compare associations among multimorbid conditions by age and sex in a large number of middle-aged and older Chinese adults. METHODS: Data from the home pages of inpatient medical records in the Shenzhen National Health Information Platform were evaluated. From January 1, 2017, to December 31, 2018, inpatients aged 50 years and older who had been diagnosed with at least one of 40 conditions were included in this study. Their demographic characteristics (age and sex) and inpatient diagnoses were extracted. Association rule mining, Chi-square tests, and decision tree analyses were combined to identify associations between multiple chronic conditions. RESULTS: In total, 306,264 hospitalized cases with available information on related chronic conditions were included in this study. The prevalence of multimorbidity in the overall population was 76.46%. The combined results of the 3 analyses showed that, in patients aged 50 years to 64 years, lipoprotein metabolism disorder tended to be comorbid with multiple chronic conditions. Gout and lipoprotein metabolism disorder had the strongest association. Among patients aged 65 years or older, there were strong associations between cerebrovascular disease, heart disease, lipoprotein metabolism disorder, and peripheral vascular disease. The strongest associations were observed between senile cataract and glaucoma in men and women. In particular, the association between osteoporosis and malignant tumor was only observed in middle-aged and older men, while the association between anemia and chronic kidney disease was only observed in older women. CONCLUSIONS: Multimorbidity was prevalent among middle-aged and older Chinese individuals. The results of this comprehensive analysis of 4 age-sex subgroups suggested that associations between particular conditions within the sex and age groups occurred more frequently than expected by random chance. This provides evidence for further research on disease clusters and for health care providers to develop different strategies based on age and sex to improve the early identification and treatment of multimorbidity.

Combination of AIBP, apoA-I, and Aflibercept Overcomes Anti-VEGF Resistance in Neovascular AMD by Inhibiting Arteriolar Choroidal Neovascularization.

Purpose: Anti-VEGF resistance represents a major unmet clinical need in the management of choroidal neovascularization (CNV). We have previously reported that a combination of AIBP, apoA-I, and an anti-VEGF antibody overcomes anti-VEGF resistance in laser-induced CNV in old mice in prevention experiments. The purpose of this work is to conduct a more clinically relevant study to assess the efficacy of the combination of AIBP, apoA-I, and aflibercept in the treatment of anti-VEGF resistance of experimental CNV at different time points after laser photocoagulation. Methods: To understand the pathobiology of anti-VEGF resistance, we performed comprehensive examinations of the vascular morphology of laser-induced CNV in young mice that are highly responsive to anti-VEGF treatment, and in old mice that are resistant to anti-VEGF therapy by indocyanine green angiography (ICGA), fluorescein angiography (FA), optical coherence tomography (OCT), and Alexa 568 isolectin labeled choroid flatmounts. We examined the efficacy of the combination therapy of AIBP, apoA-I, and aflibercept intravitreally delivered at 2, 4, and 7 days after laser photocoagulation in the treatment of CNV in old mice. Results: Laser-induced CNV in young and old mice exhibited cardinal features of capillary and arteriolar CNV, respectively. The combination therapy and the aflibercept monotherapy were equally effective in treating capillary CNV in young mice. In old mice, the combination therapy was effective in treating anti-VEGF resistance by potently inhibiting arteriolar CNV, whereas aflibercept monotherapy was ineffective. Conclusions: Combination therapy of AIBP, apoA-I, and aflibercept overcomes anti-VEGF resistance in experimental CNV in old mice by inhibiting arteriolar CNV.

[Regulation of PARP-1 deficiency on epidermal growth factor receptor during lung injury of mice induced by benzo [a] pyrene inhalation exposure].

OBJECTIVE: To investigate the regulation of PARP-1 deficiency on epidermal growth factor receptor(EGFR) during lung injury of mice induced by benzo[a]pyrene(B[a]P) inhalation exposure. METHODS: PARP-1 knockout mice(PARP-1~(-/-)) and WT mice were selected as the object, and which were randomly assigned into either an intervention or a control group(n=40, half male and half female). The intervention group were individually treated with 10.0 µg/m~(3 )B[a]P for 180 days by dynamic inhalation exposure(6 h per day and 5 days per week), and the control group was given the solvent dimethyl sulfoxide(DMSO) during the same period. The expression of EGFR in lung tissues of animals were examined by RT-PCR, Western blot and immunofluorescence. RESULTS: In WT mice, the intervention manifested significant increase expression of EGFR in lung tissue, but no changes were found in the control. In PARP-1~(-/-) mice, the intervention manifested significant inhibition expression of EGFR, but the control group exhibited no changes. CONCLUSION: PARP-1 deficiency suppresses the abnormal activation of EGFR during lung injury of mice induced by B[a]P inhalation exposure.

Age differential response to bevacizumab therapy in choroidal neovascularization in rabbits.

Choroidal neovascularization (CNV) in young rabbits has been shown to have a rapid, robust response after treatment with bevacizumab, an anti-vascular endothelial growth factor (VEGF) medication. This investigation evaluates an age differential response to bevacizumab in older populations of rabbits using multimodal high resolution molecular imaging. Young (4 months old) and life span (14 months old) rabbits were given subretinal injections of Matrigel and VEGF to produce CNV. All CNV rabbit models were then treated with a bevacizumab intravitreal injection. Rabbits were then monitored longitudinally using photoacoustic microscopy (PAM), optical coherence tomography (OCT), color photography, and fluorescence imaging. Chain-like gold nanoparticle clusters (CGNP) conjugated with tripeptide arginylglycylaspartic acid (RGD) was injected intravenously for molecular imaging. Robust CNV developed in both young and old rabbits. After intravitreal bevacizumab injection, fluorescence signals were markedly decreased 90.13% in the young group. In contrast, old rabbit CNV area decreased by only 10.56% post-bevacizumab treatment. OCT images confirmed a rapid decrease of CNV in the young group. CGNPs demonstrated high PAM signal in old rabbits and minimal PAM signal in young rabbits after bevacizumab, indicating CNV regression. There is a significant difference in response to intravitreal bevacizumab treatment between young and old rabbits with CNV which can be monitored with multimodal molecular imaging. Old rabbits demonstrate significant persistent disease activity. This represents the first large eye model of persistent disease activity of CNV and could serve as the foundation for future investigations into the mechanism of persistent disease activity and the development of novel therapies.

Visual pigment-deficient cones survive and mediate visual signaling despite the lack of outer segments.

Rhodopsin and cone opsins are essential for light detection in vertebrate rods and cones, respectively. It is well established that rhodopsin is required for rod phototransduction, outer segment disk morphogenesis, and rod viability. However, the roles of cone opsins are less well understood. In this study, we adopted a loss-of-function approach to investigate the physiological roles of cone opsins in mice. We showed that cones lacking cone opsins do not form normal outer segments due to the lack of disk morphogenesis. Surprisingly, cone opsin-deficient cones survive for at least 12 mo, which is in stark contrast to the rapid rod degeneration observed in rhodopsin-deficient mice, suggesting that cone opsins are dispensable for cone viability. Although the mutant cones do not respond to light directly, they maintain a normal dark current and continue to mediate visual signaling by relaying the rod signal through rod-cone gap junctions. Our work reveals a striking difference between the role of rhodopsin and cone opsins in photoreceptor viability.

Acute effects of ambient nitrogen oxides and interactions with temperature on cardiovascular mortality in Shenzhen, China.

BACKGROUND: Though inconsistent, acute effects of ambient nitrogen oxides on cardiovascular mortality have been reported. Whereas, interactive roles of temperature on their relationships and joint effects of different indicators of nitrogen oxides were less studied. This study aimed to extrapolate the independent roles of ambient nitrogen oxides and temperature interactions on cardiovascular mortality. METHODS: Data on mortality, air pollutants, and meteorological factors in Shenzhen from 2013 to 2019 were collected. Three indicators including nitric oxide (NO), nitrogen dioxide (NO2), and nitrogen oxides (NOX) were studied. Adjusted generalized additive models (GAMs) were applied to analyse their associations with cardiovascular mortality in different groups. RESULTS: The average daily concentrations of NO, NO2, and NOX were 11.7 µg/m3, 30.7 µg/m3, and 53.2 µg/m3, respectively. Significant associations were shown with each indicator. Cumulative effects of nitrogen oxides were more obvious than distributed lag effects. Males, population under 65 years old, and population with stroke-related condition were more susceptible to nitrogen oxides. Adverse effects of nitrogen oxides were more significant at low temperature. Impacts of NO2 on cardiovascular mortality, and NO on stroke mortality were the most robust in the multi-pollutant models, whereas variations were shown in the other relationships. CONCLUSIONS: Low levels of nitrogen oxides showed acute and adverse impacts and the interactive roles of temperature on cardiovascular mortality. Cumulative effects were most significant and joint effects of nitrogen oxides required more attention. Population under 65 years old and population with stroke-related health condition were susceptible, especially days at lower temperature.

Effect of Cold Spells and Their Different Definitions on Mortality in Shenzhen, China.

A high premium has been put on researching the effects of cold spells because of their adverse influence on people's daily lives and health. The study aimed to find the most appropriate definition of the cold spell in Shenzhen and quantify the impact of cold spells on mortality. Based on the daily mortality data in Shenzhen from 2013 to 2017 and the meteorological and pollutant data from the same period, we quantified the effect of cold spells using eight different definitions in the framework of a distributed lag non-linear model with a quasi-Poisson distribution. In Shenzhen, low temperatures increase the risk of death more significantly than high temperatures (using the optimal temperature as the cut-off value). Comparing the quasi-Akaike information criterion value, attribution fraction (b-AF), and attribution number (b-AN) for all causes of deaths and non-accidental deaths, the optimal definition of the cold spell was defined as the threshold was 3rd percentile of the daily average temperature and duration for 3 or more consecutive days (all causes: b-AF = 2.31% [1.01-3.50%], b-AN = 650; non-accidental: b-AF = 1.92% [0.57-3.17%], b-AN = 471). For cardiovascular deaths, the best definition was the temperature threshold as the 3rd percentile of the daily average temperature with a duration of 4 consecutive days (cardiovascular: b-AF = 1.37% [0.05-2.51%], b-AN = 142). Based on the best definition in the model, mortality risk increased in cold spells, with a statistically significant lag effect occurring as early as the 4th day and the effect of a single day lasting for 6 days. The maximum cumulative effect occurred on the 14th day (all-cause: RR = 1.54 [95% CI, 1.20-1.98]; non-accidental: RR = 1.43 [95% CI, 1.11-1.84]; cardiovascular: RR = 1.58 [95% CI, 1.00-2.48]). The elderly and females were more susceptible to cold spells. Cold spells and their definitions were associated with an increased risk of death. The findings of this research provide information for establishing an early warning system, developing preventive measures, and protecting susceptible populations.

Years of life lost and mortality risk attributable to non-optimum temperature in Shenzhen: a time-series study.

To assess YLL and mortality burden attributable to non-optimum ambient temperature, we collected mortality and environmental data from June 1, 2012 to December 30, 2017 in Shenzhen. We applied distributed lag nonlinear models with 21 days of lag to examine temperature-YLL and temperature-mortality associations, and calculated the attributable fractions of YLL and deaths for non-optimum temperature, including four subranges, mild cold, mild heat, extreme cold, and extreme heat. Cold and heat were distinguished by the optimum temperature, and each was separated into extreme and mild by cutoffs at 2.5th (12.2 °C) and 97.5th (30.4 °C) temperature percentile further. The optimum temperature was defined as the temperature that had minimum effect on YLL or mortality risk. The optimum temperature for non-accidental YLL was 24.5 °C, and for mortality it was 25.4 °C. Except for the population older than 65 years, the optimum temperature was generally lower in the YLL model than the mortality model. Of the total 61,576 non-accidental deaths and 1,350,835.7 YLL within the study period, 17.28% (95% empirical CI 9.42-25.14%) of YLL and 17.27% (12.70-21.34%) of mortality were attributable to non-optimum temperature. More YLL was caused by cold (10.14%, 3.94-16.36%) than by heat (7.14%, 0.47-13.88%). Mild cold (12.2-24.5 °C) was responsible for far more YLL (8.78%, 3.00-14.61%) than extreme cold (3.5-12.2 °C). As for cardiovascular deaths, only the fractions attributable to overall and cold temperature were significant, with mild cold contributing the largest fraction to YLL (16.31%, 6.85-25.82%) and mortality (16.08%, 9.77-21.22%). Most of the temperature-related YLL and mortality was attributable to mild but non-optimum weather, especially mild cold, while the YLL model implied a more prominent heat effect on premature death. Our findings can supply additional evidence from multiperspectives for health planners to define priorities and make targeted policies for mitigating the burden of adverse temperatures.

Combination of apolipoprotein-A-I/apolipoprotein-A-I binding protein and anti-VEGF treatment overcomes anti-VEGF resistance in choroidal neovascularization in mice.

Many patients of choroidal neovascularization (CNV) are unresponsive to the current anti-VEGF treatment. The mechanisms for anti-VEGF resistance are poorly understood. We explore the unique property of the apolipoprotein A-I (apoA-I) binding protein (AIBP) that enhances cholesterol efflux from endothelial cells and macrophages to thereby limit angiogenesis and inflammation to tackle anti-VEGF resistance in CNV. We show that laser-induced CNV in mice with increased age showed increased resistance to anti-VEGF treatment, which correlates with increased lipid accumulation in macrophages. The combination of AIBP/apoA-I and anti-VEGF treatment overcomes anti-VEGF resistance and effectively suppresses CNV. Furthermore, macrophage depletion in old mice restores CNV sensitivity to anti-VEGF treatment and blunts the synergistic effect of combination therapy. These results suggest that cholesterol-laden macrophages play a critical role in inducing anti-VEGF resistance in CNV. Combination therapy by neutralizing VEGF and enhancing cholesterol removal from macrophages is a promising strategy to combat anti-VEGF resistance in CNV.

[Establishment of mouse lung cancer model induced by benzo[a]pyrene dynamic inhalation exposure].

OBJECTIVE: To establish a mouse lung cancer model induced by benzo[a]pyrene(B[a]P) dynamic inhalation exposure. METHODS: A total of 96 C57 BL/6 J mice weighing 18 to 20 g were randomly divided into the control group(n=48)and the experimental group(n=48), male and female in half. The experimental group was treated with 10. 0 µg/m~3 BaP for 13 weeks or 25 weeks(6 h per day and 5 days per week) by dynamic inhalation exposure, while the control group was given dimethyl sulfoxide(DMSO). The 13 weeks or 25 weeks after B[a]P exposure, 24 mice were sacrificed and their lung tissues dissected and observed for tumor formation. During B[a]P exposure, the gas in the poisoning cabinet was collected by the active carbon tube method regularly, and the concentration of B[a]P in the poisoning cabinet was analyzed by gas chromatography mass spectrometer, and the(7 R, 8 S)-dihydroxy-(9 S, 10 R)-epoxy-7, 8, 9, 10-tetrahydrobenzo [a] pyrene(BPDE)-DNA adduct content in the whole blood of mice was determined by ELISA. RESULTS: Concentration of B[a]P in the dynamic inhalation cabinet was stable at(12. 794±0. 518)µg/m~3. There was no significant difference in the BPDE-DNA content in the experimental objects between 13 weeks and 25 weeks after B[a]P exposure(P>0. 05). After 25 weeks of B[a]P exposure, the lung cancer formation rate in the experimental group was significantly higher than that in the control group(P<0. 05), the tumor formation rate of female mice was up to 100%. CONCLUSION: The mice lung cancer model induced by B[a]P dynamic inhalation exposure was established successfully.

Effect of temperature on accidental human mortality: A time-series analysis in Shenzhen, Guangdong Province in China.

Health-risk assessments of temperature are central to determine total non-accidental human mortality; however, few studies have investigated the effect of temperature on accidental human mortality. We performed a time-series study combined with a distributed lag non-linear model (DLNM) to quantify the non-linear and delayed effects of daily mean temperature on accidental human mortality between 2013 and 2017 in Shenzhen, China. The threshold for effects of temperature on accidental human mortality occurred between 5.6 °C and 18.5 °C. Cold exposures, but not hot exposures, were significantly associated with accidental human mortality. All of the observed groups were susceptible to cold effects, with the strongest effects presented in females (relative risk [RR]: 3.14, 95% confidence interval (CI) [1.44-6.84]), followed by poorly educated people (RR: 2.63, 95% CI [1.59-4.36]), males (RR: 1.79, 95% CI [1.10-2.92]), and well-educated people (RR: 1.20, 95% CI [0.58-2.51]). Pooled estimates for cold effects at a lag of 0-21 days (d) were also stronger than hot effects at a lag of 0-2 d. Our results indicate that low temperatures increased the risk of accidental human mortality. Females and poorly educated people were more susceptible to the low temperatures. These findings imply that interventions which target vulnerable populations during cold days should be developed to reduce accidental human mortality risk.

Deletion of M-Opsin Prevents M Cone Degeneration in a Mouse Model of Leber Congenital Amaurosis.

Mutations in retinoid isomerase (RPE65) or lecithin-retinol acyltransferase (LRAT) disrupt 11-cis-retinal synthesis and cause Leber congenital amaurosis (LCA). Despite the success of recent RPE65 gene therapy, follow-up studies show that patients continue to experience photoreceptor degeneration and lose vision benefit over time. In Lrat-/- mouse model, mislocalized medium (M)-wavelength opsin was degraded, whereas mislocalized short (S)-wavelength opsin accumulated before the onset of cone degeneration. The mechanism for the foveal M/long-wavelength cone photoreceptor degeneration in LCA is unknown. By crossing Lrat-/- mice with a proteasome reporter mouse strain, this study showed that M-opsin-enriched dorsal cones in Lrat-/- mice exhibit proteasome stress because of the degradation of large amounts of M-opsin. Deletion of M-opsin relieves the proteasome stress and completely prevents M cone degeneration in Lrat-/-Opn1sw-/- mice (a pure M cone LCA model, Opn1sw encoding S-opsin) for at least 12 months. These results suggest that M-opsin degradation-associated proteasome stress plays a major role in M cone degeneration in Lrat-/- model. This finding may represent a general mechanism for M cone degeneration in multiple forms of cone degeneration because of M-opsin mislocalization and degradation. These results have important implications for the current gene therapy strategy for LCA that emphasizes the need for combinatorial therapies to both improve vision and slow photoreceptor degeneration.

Author Correction: Characterization of Retinal Ganglion Cell and Optic Nerve Phenotypes Caused by Sustained Intracranial Pressure Elevation in Mice.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Effect of changes in season and temperature on cardiovascular mortality associated with nitrogen dioxide air pollution in Shenzhen, China.

BACKGROUND: The intricate association of mortality risk with ambient air pollution and temperature is of growing concern. Little is known regarding effect of changes in season and temperature on daily cardiovascular mortality associated with air pollutant nitrogen dioxide (NO2). OBJECTIVES: Our study aimed to assess the effect of NO2 on cardiovascular mortality modified by season and daily air temperature in the effect, and further to identify the population highly susceptible to cardiovascular mortality associated with NO2 and air temperature. METHODS: We collected daily cause-specific death data, weather conditions, and air pollutant concentrations in Shenzhen from 2013 to 2017. Distributed-lag linear models were employed to analyze the effect of season on the NO2-associated mortality. Furthermore, generalized additive models were combined with stratification parametric analysis to estimate the interaction effect of NO2 with air temperature on cardiovascular mortality. RESULTS: In the cold season, the percentage increase in daily mortality for every 10 µg/m3 increment in NO2 concentration over lags of 0-2 days was 4.45% (95% CI: 2.71-6.21%). However, no statistically significant effect of NO2 was observed in the warm season. Compared with high-temperature days (>median temperature), a 3.51% increase in mortality (95% CI: 2.04-5.01%) over low-temperature days (≤median temperature) for the same increase in NO2 was significant. Air temperature modified the effect of NO2 on daily mortality by 4.08% (95% CI: 2.28-5.91%) for the elderly (age ≥ 65 years) on low-temperature days vs. -0.82% (95% CI: -3.88-2.34%) on high-temperature days, and 3.38% (95% CI: 1.50-5.29%) for males on low-temperature days vs. -0.73% (95% CI: -3.83-2.47%) on high air temperature days. CONCLUSIONS: The cold season and low temperatures could significantly enhance the effect of NO2 on cardiovascular mortality. The elderly and males suffering from cardiovascular disease should take precautions against low temperature and NO2 air pollution.

Regulation of Wnt Singaling Pathway by Poly (ADP-Ribose) Glycohydrolase (PARG) Silencing Suppresses Lung Cancer in Mice Induced by Benzo(a)pyrene Inhalation Exposure.

Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon that specifically causes cancer and is widely distributed in the environment. Poly (ADP-ribosylation), as a key post-translational modification in BaP-induced carcinogenesis, is mainly catalyzed by poly (ADP-ribose) glycohydrolase (PARG) in eukaryotic organisms. Previously, it is found that PARG silencing can counteract BaP-induced carcinogenesis in vitro, but the mechanism remained unclear. In this study, we further examined this process in vivo by using heterozygous PARG knockout mice (PARG+/-). Wild-type and PARG+/- mice were individually treated with 0 or 10 µg/m3 BaP for 90 or 180 days by dynamic inhalation exposure. Pathological analysis of lung tissues showed that, with extended exposure time, carcinogenesis and injury in the lungs of WT mice was progressively worse; however, the injury was minimal and carcinogenesis was not detected in the lungs of PARG+/- mice. These results indicate that PARG gene silencing protects mice against lung cancer induced by BaP inhalation exposure. Furthermore, as the exposure time was extended, the protein phosphorylation level was down-regulated in WT mice, but up-regulated in PARG+/- mice. The relative expression of Wnt2b and Wnt5b mRNA in WT mice were significantly higher than those in the control group, but there was no significant difference in PARG+/- mice. Meanwhile, the relative expression of Wnt2b and Wnt5b proteins, as assessed by immunohistochemistry and Western blot analysis, was significantly up-regulated by BaP in WT mice; while in PARG+/- mice it was not statistically affected. Our work provides initial evidence that PARG silencing suppresses BaP induced lung cancer and stabilizes the expression of Wnt ligands, PARG gene and Wnt ligands may provide new options for the diagnosis and treatment of lung cancer.

Association between PM2.5 Exposure and All-Cause, Non-Accidental, Accidental, Different Respiratory Diseases, Sex and Age Mortality in Shenzhen, China.

Background: China is at its most important stage of air pollution control. Research on the association between air pollutants and human health is very important and necessary. The purpose of this study was to evaluate the association between PM2.5 concentrations and residents' mortality and to compare the effect of PM2.5 on the different diseases, accidental deaths, sex or age of residents from high polluted areas with less polluted areas. Methods: The semi-parametric generalized additive model (GAM) with Poisson distribution of time series analysis was used. The excess risk (ER) of mortality with the incremental increase of 10 µg/m³ in PM2.5 concentration was calculated. Concentration-response relationship curves and autocorrelation between different lags of PM2.5 were also evaluated. Results: PM2.5 exposure was significantly associated with the mortality of residents. The strongest ERs per 10 µg/m³ increase in PM2.5 were 0.74% (95% CI: 0.11⁻1.38%) for all-cause, 0.67% (95% CI: 0.01⁻1.33%) for non-accidental, 1.81% (95% CI: 0.22⁻3.42%) for accidental, 3.04% (95% CI: 0.60⁻5.55%) for total respiratory disease, 6.38% (95% CI: 2.78⁻10.11%) for chronic lower respiratory disease (CLRD), 8.24% (95% CI: 3.53⁻13.17%) for chronic obstructive pulmonary disease (COPD), 1.04% (95% CI: 0.25⁻1.84%) for male and 1.32% (95% CI: 0.46⁻2.19%) for elderly. Furthermore, important information on the concentration-response relationship curves was provided. Conclusions: PM2.5 can increase the risk of residents' mortality, even in places with less air pollution and developed economy in China.