Orientin alleviates ox-LDL-induced oxidative stress, inflammation and apoptosis in human vascular endothelial cells by regulating Sestrin 1 (SESN1)-mediated autophagy.

Endothelial cells are a crucial component of the vessel-tissue wall and exert an important role in atherosclerosis (AS). To explore the role of Orientin in AS, human vascular endothelial cells (HUVECs) were induced by oxidized low-density lipoprotein (ox-LDL) to simulate the vascular endothelial injury during AS. Cell viability was detected by CCK-8 assay. Oxidative stress and inflammation related markers were measured using kits, RT-qPCR or western blot. Besides, cell apoptosis was assessed with TUNEL staining and cell autophagy was evaluated by LC3 immunofluorescent staining. Additionally, western blot was utilized to evaluate the expression of Sestrin 1 (SESN1) and proteins in AMPK/mTOR signaling. Afterwards, SESN1 was silenced to determine the expression of autophagy-related proteins. The further application of autophagy inhibitor 3-methyladenine (3-MA) was used to clarify the regulatory mechanism of Orientin on autophagy. Results showed that the decreased viability of HUVECs caused by ox-LDL induction was elevated by Orientin. Oxidative stress and inflammation were also attenuated after Orientin addition in HUVECs under ox-LDL condition. Moreover, Orientin suppressed apoptosis and induced autophagy of HUVECs stimulated by ox-LDL, accompanied by enhanced level of phospho (p)-AMPK and declined level of p-mTOR. Interestingly, SESN1 level was elevated by Orientin, and SESN1 depletion alleviated autophagy and reduced p-AMPK expression but enhanced p-mTOR expression. The further experiments indicated that SESN1 silencing or 3-MA addition reversed the inhibitory effects of Orientin on the oxidative stress, inflammation and apoptosis of HUVECs. Collectively, Orientin could induce autophagy by activating SESN1 expression, thereby regulating AMPK/mTOR signaling in ox-LDL-induced HUVECs.

Suppression of lncRNA Gm47283 attenuates myocardial infarction via miR-706/ Ptgs2/ferroptosis axis.

Myocardial infarction (MI) is the leading cause of sudden death. Long non-doing RNAs (lncRNAs) were demonstrated to play crucial roles in multiple diseases, including cancer and cardiovascular diseases. Nevertheless, the molecular mechanism of lncNRAs in MI is unclear. In this study, we integrated bioinformatics and molecular biological experiments to identify the novel lncRNA transcripts and elucidated its regulatory mechanism in MI. First, we identified 10 dysregualted lncRNAs and found that lncRNA Gm47283 was the top risk factor in MI. Bioinformatics analysis predicted that lncRNA Gm47283 exerted function via targeting miR-706 and Ptgs2. Ptgs2 was also the known regulator of ferroptosis. Inhibition or overexpression of lncRNA Gm47283 could regulate Ptgs2 expression and downstream ferroptosis activity. Overexpression of miR-706 could inhibit the expression of Ptgs2 and the activity of ferroptosis, thereby attenuated cellular injury. Mechanically, co-transfection experiments showed that overexpression of miR-706 could reverse the damage effect that was caused by lncRNA Gm47283 overexpression, via inhibiting Ptgs2 and ferroptosis. Additionally, inhibition of lncRNA Gm47283 by stem cell membrane coated siRNA could attenuate MI in vivo. Our study elucidated a novel mechanism containing lncRNA Gm47283/miR-706/Ptgs2/ferroptosis in MI, which provided a potential therapeutic for MI.Graphical Abstract. Stem cell membrane coated siRNA of lncRNA Gm47283 inhibits cardiomyocyte ferroptosis in myocardial infarction rat. Stem cell membrane-coated siRNA of lncRNA Gm47283 increases miR-706, and then miR-706 suppresses the expression of Ptgs2 to reduce lipid peroxidation toxicity, and then inhibits cardiomyocyte ferroptosis. PUFA: polyunsaturated fatty acid.

Mitochondrial targeted astaxanthin liposomes for myocardial ischemia-reperfusion injury based on oxidative stress.

Myocardial ischemia-reperfusion injury (MI/RI) refers to the clinical state of decreased coronary blood flow caused by various causes. The main pathogenesis of MI/RI is mitochondrial oxidative damage. In this study, we designed a novel mitochondrial targeted astaxanthin (AST) liposome, namely, STPP-AST-LIP, targeting mitochondria of H9c2 myocardial cells. STPP-AST-LIP not only reduced the production of mitochondrial reactive oxygen species (ROS), but also increased the survival rate of MI/RI H9c2 cells. In addition, rat experiments further confirmed that STPP-AST-LIP could improve myocardial cardiac function in MI/RI rats, significantly inhibited apoptosis of myocardial cells, and had a protective effect on the heart of rats after MI/RI.

SESN1 attenuates the Ox­LDL­induced inflammation, apoptosis and endothelial­mesenchymal transition of human umbilical vein endothelial cells by regulating AMPK/SIRT1/LOX1 signaling.

Endothelial cells are an important component of the heart and vasculature and form a crucial link between the cardiovascular system and the immune system. Sestrin 1 (SESN1) has an important role in atherosclerosis by inhibiting NOD­like receptor family pyrin domain containing 3 inflammasome activation. However, whether SESN1 is involved in human umbilical vein endothelial cell (HUVEC) injury caused by atherosclerosis has remained to be elucidated. The present study aimed to investigate the functions of SESN1 in the inflammatory response, apoptosis and endothelial­mesenchymal transition (EndMT) of HUVECs following stimulation with oxidized low­density lipoprotein (Ox­LDL). SESN1 expression at the mRNA and protein levels was detected using reverse transcription­quantitative PCR (RT­qPCR) and western blot analysis. Following SESN1 overexpression in Ox­LDL­stimulated HUVECs, cell viability was determined using a Cell Counting Kit­8 assay. Terminal deoxynucleotidyl transferase­mediated nick­end labeling staining was employed to detect cell apoptosis and western blot analysis was used to determine the levels of apoptosis­related proteins. RT­qPCR, ELISA and western blot were utilized to determine the levels of inflammatory factors. Immunofluorescence staining, RT­qPCR and western blot analysis were employed to assess the EndMT of Ox­LDL­stimulated HUVECs. The results revealed that SESN1 exhibited a low expression in HUVECs following Ox­LDL stimulation. SESN1 overexpression suppressed inflammation, apoptosis and EndMT in Ox­LDL­induced HUVECs. In addition, SESN1 stimulated adenosine monophosphate­activated protein kinase catalytic subunit α1/sirtuin 1 signaling to suppress Ox­LDL receptor­1 expression. An AMPK and SIRT1 inhibitor reversed the effects of SESN1 overexpression on the inflammatory response, apoptosis and EndMT of HUVECs exposed to Ox­LDL. Taken together, the present study demonstrated that SENS1 exerts a suppressive effect on Ox­LDL­induced inflammation, apoptosis and EndMT of HUVECs, suggesting that SENS1 may be used as a novel biomarker for endothelial injury­related disorders.

Forkhead box protein 1 transcriptionally activates sestrin1 to alleviate oxidized low-density lipoprotein-induced inflammation and lipid accumulation in macrophages.

Transcription factor forkhead box protein 1 (FOXP1) has been shown cardiovascular protection. We aimed to analyze the role of FOXP1 in oxidized low-density lipoprotein (ox-LDL)-induced macrophages and its possible regulatory effect on sestrin1 (SESN1) expression. After stimulation with ox-LDL, FOXP1 expression in RAW264.7 cells was evaluated with RT-qPCR and Western blotting. Then, FOXP1 was overexpressed, followed by detection of inflammatory mediator levels using ELISA kits and RT-qPCR. Lipid accumulation was detected with oil red O staining. Additionally, the JASPAR database was used to predict the potential genes that could be transcriptionally regulated by FOXP1. ChIP and luciferase reporter assays were used to verify this combination. To further clarify the regulatory effects of FOXP1 on SESN1 in damage of macrophages triggered by ox-LDL, SESN1 was silenced to determine the inflammation and lipid accumulation under the condition of FOXP1 overexpression. Results indicated that ox-LDL stimulation led to a significant decrease in FOXP1 expression. FOXP1 overexpression notably reduced the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6, accompanied by a decreased in phosphorylated NF-κB p65 expression. Besides, FOXP1-upregulation inhibited lipid accumulation and reduced CD36 expression level in RAW264.7 cells upon ox-LDL stimulation. Moreover, results of ChIP and luciferase reporter assays suggested that FOXP1 could transcriptionally regulate SESN1 expression. Further experiments supported that SESN1 silencing restored the inhibitory effects of FOXP1 overexpression on the inflammation and lipid accumulation in RAW264.7 cells exposed to ox-LDL. Collectively, FOXP1 transcriptionally activates SESN1 for the alleviation of ox-LDL-induced inflammation and lipid accumulation in macrophages.

Do Herbaceous Species Functional Groups Have a Uniform Pattern along an Elevation Gradient? The Case of a Semi-Arid Savanna Grasslands in Southern Ethiopia.

Knowledge of the total (overall) and individual herbaceous vegetation species relating to a distinctive site might help in the development of management strategies for a large number of threatened herbaceous species. This paper assesses the total and functional group herbaceous biomass, species richness, evenness, and diversity at four elevation classes in Borana rangelands of arid thorn bush savanna grasslands in Southern Ethiopia. At each elevation class, a grid of 20 × 20 m main plot was placed, and individual herbaceous species samples were collected randomly from five 1 m2 quadrants within the main plot. Using a single-factor analysis of variance (ANOVA), the effects of four elevation classes were considered on whole-vegetation, grasses, graminoid, and forb species diversity, evenness, richness, and biomass. A total of 49 herbaceous species were recorded. Of the total identified herbaceous species, three grass species and two graminoid species were found across all studied elevation classes, but the forb species did not overlap along the studied elevation classes. The total richness, diversity, and evenness of herbaceous species were considerable and significant along elevation classes. The grass, graminoid, and forb species richness, diversity, and evenness responded differently, and the functional group of species may be a good indicator of the community processes of grassland across elevation classes. The contribution of forb richness to the total richness was more pronounced than grass and graminoid, which indicates the shift of savanna grassland to grazing tolerant herbaceous species. The results suggest that the pooled data analysis of herbaceous vegetation community structure and biomass could obscure complicate trends of the functional group at elevation classes and for managing herbaceous species in savanna grasslands, the management models should focus on the functional group species composition, community structure, and biomass.

Solid cancer incidence among Chinese medical diagnostic x-ray workers, 1950-1995: Estimation of radiation-related risks.

The objective of this study was to estimate solid cancer risk attributable to long-term, fractionated occupational exposure to low doses of ionizing radiation. Based on cancer incidence for the period 1950-1995 in a cohort of 27,011 Chinese medical diagnostic X-ray workers and a comparison cohort of 25,782 Chinese physicians who did not use X-ray equipment in their work, we used Poisson regression to fit excess relative risk (ERR) and excess absolute risk (EAR) dose-response models for incidence of all solid cancers combined. Radiation dose reconstruction was based on a previously published method that relied on simulating measurements for multiple X-ray machines, workplaces and working conditions, information about protective measures, including use of lead aprons, and work histories. The resulting model was used to estimate calendar year-specific badge dose calibrated as personal dose equivalent (Sv). To obtain calendar year-specific colon doses (Gy), we applied a standard organ conversion factor. A total of 1,643 cases of solid cancer were identified in 1.45 million person-years of follow-up. In both ERR and EAR models, a statistically significant radiation dose-response relationship was observed for solid cancers as a group. Averaged over both sexes, and using colon dose as the dose metric, the estimated ERR/Gy was 0.87 (95% CI: 0.48, 1.45), and the EAR was 22 per 10(4)PY-Gy (95% CI: 14, 32) at age 50. We obtained estimates of the ERR and EAR of solid cancers per unit dose that are compatible with those derived from other populations chronically exposed to low dose-rate occupational or environmental radiation.

[Survey and analysis of awareness of lung cancer prevention and control in a LDCT lung cancer screening project in Tianjin Dagang Oilfield of China].

BACKGROUND AND OBJECTIVE: It has been proven that increase of the awareness level of lung cancer prevention and control could enhance participation of lung cancer screening of lung cancer high risk group. The aim of this study is to investigate the awareness level of lung cancer prevention and control and the effect of individual characteristics on lung cancer awareness, and to provide evidence for comprehensive lung cancer prevention in high risk areas of lung cancer. METHODS: Staffs of Tianjin Dagang Oil Field who participate low dose CT (LDCT) lung cancer screening by cluster sampling or according to voluntary principle were surveyed, data of lung cancer awareness were collected by questionnaire. RESULTS: A total of 1,633 valid questionnaires were collected. The average age of respondents was 60.08±6.58. Most participants were males (82.2%) while female only accounted for 17.8%. The proportions of awareness about lung cancer in China, risk factors, screening methods and the knowledge of health examination were 64.5%, 77.1%, 43.7%, 49.6% respectively. Result of multiple logistic regression analysis showed that education level, smoking (pack-year), age, prior tuberculosis were the influencing factors of lung cancer awareness with adjusted Ors for education and age level as of 0.567 (95%CI: 0.439-0.733) and 1.373 (95%CI: 1.084-1.739) respectively. 80.3% of the participants can accept health examination once a year, while the ability to pay the medical expenses was not high. The influencing factors of health examination willingness were gender, age, income, the knowledge of lung cancer. CONCLUSIONS: Education level and smoking affect the awareness of lung cancer prevention and control, health education for lung cancer should be conducted especially in population with low education level. Comprehensive lung cancer control in high risk areas should combined lung cancer screening, tobacco control and health education.

[Advance of lung cancer screening with low-dose spiral CT].

Lung cancer has become the leading cause of cancer mortality globally, and 5-year survival rate is very poor. Screening and early detection are vital to improve survival and decrease mortality of lung cancer. In recent 20 years, low-dose spiral CT (LDCT) screening has become a research focus in this area. Randomized controlled trials have confirmed that LDCT can decrease lung cancer mortality. However, there are still some problems of LDCT. In this paper, we summarized the controversy that whether low-dose helical CT screening can reduce lung cancer mortality or not before its effectiveness was been confirmed, the results and problems in the randomized controlled trials and gave a prospect of low-dose helical CT screening's future application.

[A cohort study on risk factors of lung cancer in Yunnan tin miners].

BACKGROUND: Smoking is a major cause of lung cancer. Studies of lung cancer among miners have shown that occupational exposure also played an important role. The aim of this study is to investigate radon, cigarette use and other risk factors of lung cancer in Yunnan tin miners and to provide a scientific basis for the prevention and control of occupational lung cancer. METHODS: A prospective cohort study was conducted among Yunnan tin miners, the associations between potential risk factors for lung cancer were analyzed by multivariate Cox regression model. Effects of age at first radon exposure and radon exposure rate on lung cancer risk were analyzed. The relationship between cumulative working level month and lung cancer was analyzed according to smoking status. The joint effect of tobacco use and cumulative radon exposure was analyzed based on additive and multiplicative models. RESULTS: Increased risk of lung cancer was associated with age at enrollment, tobacco use, prior bronchitis, and cumulative arsenic and radon exposure, while higher education level was associated with decreased lung cancer risk. An inverse effect of radon exposure rate was observed. There was no significant association between lung cancer risk and first radon exposure age. There was a significant additive interaction between tobacco use and radon exposure on lung cancer risk. CONCLUSIONS: Several risk factors may contribute to the high incidence of lung cancer in Yunnan tin miners. Further studies are warranted to evaluate joint effect of different risk factors.

Effects of gestational cadmium exposure on pregnancy outcome and development in the offspring at age 4.5 years.

The objective of the present study was to evaluate the potential effect of maternal cadmium exposure on pregnancy outcome and development in the offspring at age 4.5 years. Between November 2002 and December 2003, 109 normal pregnant women were enrolled in our cohort from Da-Ye Country, Hubei Province in Central China. The placental, whole blood, and cord blood levels of cadmium were determined by inductively coupled plasma mass spectrometer (ICP-MS). The 106 children at 4.5 years of age given birth by the aforementioned women were followed up and the following rate was 97.25%. Detailed questionnaire surveys, anthropometric measurements were performed, and IQ development was evaluated by Wechsler Preschool and Primary Scale of Intelligence Revised Edition (WPPSI-R). Multiple linear regression analysis indicated that cord blood cadmium level was significantly negatively correlated with fetus development. Low birth weight (less than 2,500 g) occurred significantly more frequently in infants with higher cord blood cadmium than in those exposed to lower levels of cord blood cadmium. Significantly negative correlation was found between cord blood cadmium exposure and WPPSI-R IQ full score after controlling for confounding variables. It was concluded that cord blood cadmium concentration was a factor that influenced fetus growth and later IQ development.

[A case control study of lung cancer among workers in dagang oil field, tianjin, china.].

BACKGROUND: To investigate the risk factors of lung cancer among the employees who had worked in Dagang oil field, Tianjin, China. METHODS: A matched case control study was conducted to analyse the association between some exposures and lung cancer risk by univariate and multiple Logistic regression method. RESULTS: According to the results from univariate and multiple analysis, smoking and previous lung disease increase lung cancer risk with adjusted OR of 1.52 (95%CI: 1.18-1.94) and 3.37 (95%CI: 1.88-7.16) respectively, while the adjusted OR for occupational chemical toxic substance and dust exposure is 0.73 (95%CI: 0.69-1.30) and 0.84 (95%CI: 0.62-1.15) respectively, and there is no significant association between family history of cancer and lung cancer risk in this study. CONCLUSIONS: Smoking and previous lung disease are the independent risk factors for lung cancer among workers in Dagang oil field, yet due to some potential epidemiological bias, the association between occupational exposure and lung cancer needs further investigation.

[Factors related to Glycophorin A mutation frequency of workers exposed to benzene.].

OBJECTIVE: To analyze related factors which affect GPA mutation frequency of workers exposed to benzene, with the Glycophorin A (GPA) mutation assay and explore the possibility of GPA mutation frequency as an index of predicting the risk of benzene poisoning. METHODS: The erythrocytes were bound with fluorescent-labeled monoclonal antibody after isolated and fixed from the peripheral blood, and then the GPA mutation assay was performed using the flow cytometry (FCM). The related factors of GPA mutation frequency were analyzed by statistical methods. RESULTS: The GPA mutation frequency of chronic benzene poisonings was significantly higher than that of their controls (P < 0.05). Significant direct correlation was found between age, length of service, accumulative exposure score and the GPA mutation frequency of workers exposed to benzene (P < 0.01). However, there was significantly inverse correlation between the 3AB index and the GPA mutation frequency (GPAN0: r(s) = -0.589, P < 0.01, GPANN: r(s) = -0.615, P < 0.01). In the multiple factor regression analysis on GPA mutation frequency, benzene exposure and individual susceptibility both entered model of multiple factors analysis, the coefficient of determination of benzene-exposed workers was 0.819. CONCLUSION: Exposure to benzene and individual susceptibility are the most important factors that affect GPA mutation frequency. GPA mutation frequency increases with the benzene exposure and individual susceptibility.

[The application of single-cell gel electrophoresis to the diagnosis of fanconi anemia].

OBJECTIVE: To explore the feasibility of single-cell gel electrophoresis (SCGE) as one of lab tests to examine DNA breakage for the diagnosis of Fanconi anemia (FA). Case Record A 4-year-and-10-month old boy presented with cryptorchism, deformities of both thumbs and esotropia of right eye. He developed thrombocytopenia and anemia when he was 3 year- and -2-month old. He was clinically diagnosed as FA. METHODS AND RESULTS: DNA breakage of peripheral white blood cells from the patient and his parents was examined with SCGE. The percentages of cells with chromosome breakage (comet-tail positive cells) were 100%, 90% and 52% for the patient,his father and mother, respectively, while that were only 2% and 5% in two normal same-age children (P <0. 001). The micronucleus-positive lymphocytes was 6.74% in the patient, being also much higher than normal value (0.40%). CONCLUSION: SCGE disclosed DNA breakage in the patient with FA, suggesting that it could be used as a test for determining DNA breakage of FA.

[Detection of DNA injury and repair with single cell gel electrophoresis radiation in biodosimetry].

OBJECTIVE: To explore the relationship between DNA repair in vitro and in vivo after irradiation, and to describe the curves of DNA repair which can improve the accuracy of radiation dose estimation. METHODS: The DNA double-strand break in lymphocytes of human and mouse was detected using neutral single cell gel electrophoresis (SCGE) after radiation and the curves of DNA repair individually were estimated, which were compared later. RESULTS: Along with the time lapsing, the DNA repair of human peripheral blood and mice increased significantly and the residual damage decreased gradually, which showed significant time-effect relationship. The curve of DNA repair in vitro of human lymphocytes presented the same log model as that of mouse DNA repair in vivo. The curve showed as followed respectively: Mice: Y(TM) = 55.8256 - 10.792 lnX (R(2) = 0.629, P < 0.01) and Y(OTM) = 25.4173 - 4.5273 lnX (R(2) = 0.661, P < 0.01); Human: Y(TM) = 30.242 7 - 7.383 6 lnX (R(2) = 0.686, P < 0.01) and Y(OTM) = 17.9772 - 3.9125 lnX (R(2) = 0.752, P < 0.01). CONCLUSION: The curve of DNA repair in vitro of human lymphocytes could be considered in biodosimetry estimation because the process of DNA repair in vitro could display the repair level and speed of DNA double-strand break in vivo.

Effects of zinc, copper, and selenium on placental cadmium transport.

The objective of the present study was to evaluate the potential effects of zinc, copper, and selenium on placental cadmium transport. From November 2002 through January 2003, a total of 47 healthy pregnant women from Da-Ye City, Hubei Province in Central China participated in the study. Their age, parity, gestational age, pregnancy history, and lifestyle data were obtained by questionnaire interview. The placental, whole-blood, and cord blood levels of cadmium were determined by inductively coupled plasma mass spectrometer (ICP-MS), whole-blood zinc was measured by flame atomic absorption spectrometry (F-AAS), whole-blood copper by ICP-MS, and selenium was by atomic fluorescence spectrophotometry (AFS). The cord blood cadmium concentration (0.020-1.48 microg/L) was significantly lower than in maternal blood (0.80-25.20 microg/L, p<0.01). The placental cadmium concentration was from 0.082 to 3.97 microg/g dry weight. Multiple linear regression analysis indicated that lower levels of maternal blood copper were significantly associated with higher cadmium concentrations in cord blood. Placental cadmium in women with lower levels of maternal blood zinc was significantly higher than in those with normal zinc levels. The placental cadmium level in women with lower whole-blood selenium was significantly lower than in subjects with normal selenium levels. It was concluded that the essential elements copper, selenium, and zinc might significantly affect placental cadmium transport.

Effect of environmental exposure to cadmium on pregnancy outcome and fetal growth: a study on healthy pregnant women in China.

The objective of the present study was to evaluate the potential effect of environmental exposure to cadmium on pregnancy outcome and fetal growth. Normal pregnant women were selected from Da-ye city of Hubei province, a cadmium-polluted area, from November 2002 through January 2003. Whole blood of pregnant women, cord blood, and placenta were collected and cadmium levels were determined by inductively coupled plasma emission mass spectroscopy. Incidence rate of preterm labor (gestational age < or = 37 weeks) and neonatal asphyxia, neonatal birth height, and birth weight were compared between lower and higher cadmium exposure level groups. Whole blood cadmium of 44 mothers ranged from 0.80 to 25.20 microg/L. Cadmium concentration in maternal blood was significantly higher than that in cord blood (t = 11.44, P < 0.01). Placenta cadmium ranged from 0.084 to 3.97 microg/g dry weight. After adjustment for maternal age, history of gestation, abortion and lactation, Logistic regression analysis showed that there was no significant association between cadmium exposure levels and pregnancy outcome (premature labor or neonatal asphyxia). Multiple linear regression analysis showed that, cord blood cadmium level, but not maternal blood cadmium and placenta cadmium, was significantly negatively associated with neonatal birth height (t= -2.33, P < 0.05). Compared with lower cord blood cadmium level (< or = 0.40 microg/L), higher level of cord blood cadmium (>0.40 microg/L) was associated with 2.24cm decrease in neonatal birth height. There was no significant association between cadmium exposure and birth weight. It was concluded that environmental exposure to cadmium significantly lower neonatal birth height.