Pseudolaric acid B suppresses NSCLC progression through the ROS/AMPK/mTOR/autophagy signalling pathway.

Pseudolaric acid B (PAB), an acid isolated from the roots of Pseudolarix kaempferi gorden, has shown antitumour effects through multiple mechanisms of action. The objective of this study was to investigate the anticancer effect of PAB on non-small cell lung cancer (NSCLC) and its underlying mechanism. In our experiments, we observed that PAB decreased cell viability, inhibited colony formation, induced cell cycle arrest, impeded scratch healing, and increased apoptosis in H1975 and H1650 cells. Additionally, PAB treatment enhanced the fluorescence intensity of MDC staining in NSCLC cells, upregulated the protein expression of microtubule-associated protein light chain 3 II (LC3 II), and downregulated the expression of sequestosome 1 (SQSTM1/P62). Combined treatment with PAB and chloroquine (CQ) increased the protein expression levels of LC3 II and P62 while decreasing the apoptosis of H1975 and H1650 cells. Moreover, treatment with PAB led to significant mTOR inhibition and AMPK activation. PAB combined with compound C (CC) inhibited autophagy and apoptosis. Furthermore, PAB treatment increased intracellular reactive oxygen species (ROS) levels in NSCLC cells, which correlated with the modulation of the AMPK/mTOR signalling pathway and was associated with autophagy and apoptosis. Finally, we validated the antitumour growth activity and mechanism of PAB in vivo using athymic nude mice bearing H1975 tumour cells. In conclusion, our findings suggest that PAB can induce apoptosis and autophagic cell death in NSCLC through the ROS-triggered AMPK/mTOR signalling pathway, making it a promising candidate for future NSCLC treatment.

The EMT-Related Genes GALNT3 and OAS1 are Associated with Immune Cell Infiltration and Poor Prognosis in Lung Adenocarcinoma.

BACKGROUND: Lung cancer is the main cause of cancer-related death, with epithelial-mesenchymal transition (EMT) playing an important role in the development of this disease. The EMT-related genes Polypeptide N-Acetylgalactosaminyltransferase 3 (GALNT3) and 2'-5'-Oligoadenylate Synthetase 1 (OAS1) are involved in numerous tumor processes. Although these genes have been extensively studied in cancer, they have yet to be analyzed by multi-omics in lung adenocarcinoma (LUAD). METHODS: EMT-related genes were identified by R and Venn diagram. Cox regression and Kaplan-Meier analysis were performed to evaluate patient survival, and the Gene Expression Profiling Interactive Analysis (GEPIA) database was used for correlation analysis. GeneCards and R packages were used to explore gene characterization and functional annotation. The Tumor Immune Estimation Resource (TIMER), Human Protein Atlas (HPA), University of Alabama at Birmingham Cancer (UALCAN), and The Cancer Genome Atlas (TCGA) databases were used to investigate gene expression, which was then confirmed by RT-PCR. Clinicopathological analysis was carried out using the UALCAN database. Functional mechanisms and multi-omics analysis were performed using DNA Methylation Interactive Visualization Database (DNMIVD), Targetscan, TIMER, Tumor-immune System Interactions Database (TISIDB) and cBioportal. Diagnostic values were calculated using ROC curve analysis. RESULTS: A total of 320 EMT-related genes were identified in LUAD. Their characteristics were confirmed in the Database for Annotation, Visualization and Integrated Discovery (DAVID) database by the intersection of 855 and 3600 different genes from the Gene Expression Omnibus (GEO) and EMTome databases, respectively. Expression of the EMT-related genes GALNT3 and OAS1 was associated with the prognosis of LUAD patients. A positive correlation was observed between the expression of GALNT3 and OAS1, and their expression was higher in LUAD tissue than in normal lung tissue. This was confirmed using RT-PCR. Multi-omics analysis revealed that GALNT3 and OAS1 expression was associated with gene mutation and methylation, cellular immune infiltration, and several immune subtypes. A miRNA-GALNT3/OAS1 regulatory network was also found. Receiver operating characteristic (ROC) curve analysis found that GALNT3 and OAS1 expression combined had superior diagnostic value to that of each marker alone. CONCLUSIONS: GALNT3 and OAS1 expression are associated with immune cell infiltration and poor prognosis in LUAD. Their combined expression has high diagnostic value; hence, GALNT3 and OAS1 may be valuable biomarkers for the early detection of LUAD.

Organophosphorus flame retardants and their metabolites in paired human blood and urine.

Organophosphorus flame retardants (OPFRs) have been shown to be carcinogenic, neurotoxic, and endocrine disruptive, so it is important to understand the levels of OPFRs in human body as well as the modes of external exposure. In this study, we investigated the levels of 13 OPFRs and 7 phosphodiester metabolites in paired human blood and urine, as well as the influencing factors (region, age and gender), and studied the relationship between OPFRs and oxidative stress by urinary metabolites. We found that the concentrations of triphenyl phosphate (TPhP) and tris-(2-ethylhexyl) phosphate (TEHP) in the blood of urban populations were higher than those of rural populations, and that younger populations suffered higher TPhP and 2-ethylhexyl diphenyl phosphate (EHDPP) exposures than older populations. In addition, we found that tris-(2-chloroethyl) phosphate (TCEP), tributyl phosphate (TnBP), TPhP and EHDPP exposure induced oxidative stress. The results of the internal load principal component analysis indicated that dust ingestion, skin exposure, respiration and dietary intake may be the most important sources of TCEP, tris(2-butoxyethyl) phosphate (TBOEP), tri(2-chloroisopropyl) phosphate (TCIPP) and TEHP, respectively, and dust ingestion and skin exposure may be the main sources of TPhP for humans.

Organophosphate ester metabolites in human breast milk determined by online solid phase extraction coupled to high pressure liquid chromatography tandem mass spectrometry.

The analysis of metabolites of organophosphate esters (OPEs) in human breast milk is essential to evaluate OPE and OPE metabolite exposure of newborns. In the current study, an analytical method which only needs a small amount of breast milk (100 µl) was developed and validated for six diester metabolites and three hydroxylated metabolites applying salt-induced liquid-liquid extraction (SI-LLE) and dispersive solid phase extraction (d-SPE) for sample preparation and online solid phase extraction coupled to high pressure chromatography tandem mass spectrometry (online-SPE-HPLC-MS/MS) for quantitative measurement. The final method consisted of an extraction with formic acid (FA)/acetonitrile (1:200, v/v) and a cleanup with C18 d-SPE. The final extracts were trapped on a C18 cartridge with application of a wash step of 2 ml 0.1% FA milli-Q/methanol (98:2, v/v). Method detection limits (MDLs) ranging from 21.7 ng/l for BBOEHEP to 500 ng/l for BCIPP and average recoveries ranging from 58% for 5-OH-EHDPHP to 120% for BCIPP were achieved. Thirty-three breast milk samples from the LINC (Linking EDCs in maternal Nutrition to Child health) cohort collected in three distinct areas in The Netherlands were analyzed using the validated method. BCEP, BCIPP, BCIPHPP, BDCIPP, and 5-OH-EHDPHP were not detected in any of the samples, while BBOEP was the most frequently detected metabolite with a concentration range of

Occurrence and risk assessment of anthelmintics in Tuojiang River in Sichuan, China.

Recently, emerging pollutants, such as anthelmintics have attracted an increasing attention worldwide due to their extensive use and notable stability. However, the information on anthelmintics in the environment of southwest China is scarce. Thus, the occurrence, ecological risk and exposure evaluation of nineteen anthelmintics in Tuojiang River, which is one of the largest tributaries of Yangtze River, and drinking water source of Sichuan, southwest China, were investigated. The result showed that the detection frequency of anthelmintics was relatively high in Tuojiang River, ranging from 65% to 100% in river water. Among the seven kinds of anthelmintics, benzimidazoles are the primary anthelmintics, with concentrations up to 61.12 ng/L and 596.06 ng/g in water and sediment of the Tuojiang river, respectively. The total concentration of 19 anthelmintics in sediment samples from non-agricultural area was higher than that in agricultural area(p = 0.000 < 0.05). This could be attributed to anthropogenic activities, which lead to greater discharge and accumulation of anthelmintics in residential area along the river. It's worth to mention that the highest total concentrations of anthelmintics (109.28 ng/L) was found at the junction of rivers in R31 site. The results could be ascribed to the complexity of junction of Tuojiang River and Yangtze River, which could influence the distribution of pollutant. Besides, the ecological risk assessment showed that the macrocyclic lactones rather than benzimidazoles had relatively high toxicity to non-target organisms in aquatic environment (p = 0.000 < 0.05), with the highest RQEcotox value of 101 for Daphnia magna, while benzimidazoles had relatively high concentrations. The exposure risk could be ignored for both children and adults because the daily intake of anthelmintics via water ingestion were below 10 ng/kg/d. In addition, strong correlations were found between sucralose and most of the selected anthelmintics in Tuojiang River, indicating that sucralose might be a good tracer to evaluated the source of anthelmintics in surface water. This study provides the levels, risks and even some tracer information of pollutants for better understanding of anthelmintics in southwest China.

Temporal and seasonal variation and ecological risk evaluation of flame retardants in seawater and sediments from Bohai Bay near Tianjin, China during 2014 to 2017.

Seawater and sediment samples were collected from 2014 to 2017 at Bohai Bay near Tianjin, China. The median concentration of ΣOPFRs was 2202 ng/l in the seawater from 2017. ΣAlkyl-OPFRs was the predominant constitution in the seawater with a median contribution of nearly 80%, and ΣCl-OPFRs was the major component in the sediment. Regarding BFRs, BDE-209 was the principal one in the sediment. The levels of TEP, TCEP and TBEP in sediments displayed significantly seasonal variations. The summer concentration of TEP was higher than that in both the spring and autumn, and concerning TCEP and TBEP, their lowest concentration occurred in summer. The concentration of ΣOPFRs experienced a rapid increasing during 2014-2016 due to more emissions of OPFRs. The ecological risk evaluation of OPFRs and BFRs suggested a moderate and high risk to the investigated marine region under the high exposure scenario, respectively.

Grain size distribution and exposure evaluation of organophosphorus and brominated flame retardants in indoor and outdoor dust and PM10 from Chengdu, China.

To investigate overall pollution and potential risk of 32 targets (11 PBDEs, 8 NBFRs, and 13 OPFRs) in indoor and outdoor environments, dust and PM10 samples were sampled from non-occupational and occupational zones in Chengdu, China. ∑Cl-OPFRs, BDE-209, and DBDPE were the dominant constitutes in both the non-occupational and occupational dust. Regarding the PM10, no significant differences were found between the kitchen and the living room for studied OPFRs except TDCIPP, and TCEP, TCPP, TEHP, and TCP displayed significant correlation among the kitchen, the living room and personal samples. Profiles of the OPFRs in the PM10 from occupational areas presented great variation and ranked as: ∑Aryl-OPFRs < ∑Alkyl-OPFRs < ∑Cl-OPFRs. The estimated daily intakes (EDIs) of the investigated FRs via dust suggested dust ingestion and inhalation were the main exposure route to FRs, and the total of EDIs were at least one order of magnitude lower than reference data, indicating a low risk for the general public in Chengdu. However, with increasing usage of FRs in daily goods, a long-term monitoring should be conducted.

Seasonal Variation and Exposure Risks of Perchlorate in Soil, Indoor Dust, and Outdoor Dust in China.

A total of 97 paired soil, outdoor dust, and indoor dust samples were collected in the national scale of China in summer, and the perchlorate levels were compared with those in soil and outdoor dust samples collected in winter in our previous study. The median perchlorate concentrations in the outdoor dust, indoor dust, and soil samples were 8.10, 11.4, and 0.05 mg/kg, respectively, which were significantly lower than those in the winter samples due to the natural factors and human activities. No significant differences in perchlorate concentrations were found between Northern and Southern China in the dust samples, whereas the difference was obtained in the soil samples. In the terms of possible source, the perchlorate levels in the outdoor dust exhibited strong correlation with SO42- (r2 = 0.458**) and NO3- (r2 = 0.389**), indicating part of perchlorate in outdoor environment was likely from atmospheric oxidative process in summer. The perchlorate, SO42-, and Cl- levels in the indoor dust were significantly related to those in the outdoor dust, suggesting that outdoor contaminants might be an important source for indoor environment. Furthermore, the human exposure to perchlorate was under relatively safe state in China except for special sites or periods with high perchlorate levels. Dust made an unexpected contribution of 41.3% to the total daily perchlorate intake for children, whereas 2.46% for adults in China based on biomonitoring, which deserves more attention.

Concentrations and Exposure Evaluation of Metals in Diverse Food Items from Chengdu, China.

A total of 520 food samples belonging to 29 food types and 63 drinking water were collected in Chengdu market of China in 2014 to investigate the concentrations of 11 metals, and to assess the related exposure to the local consumers by estimating the hazard quotient and carcinogenic risk (CR). The results showed that metals concentrations in drinking water were below the limit values suggested by the Ministry of Health of the People's Republic of China, and FAO/WHO (Food and Agriculture Organization of the United Nations/World Health Organization). While As, Cd, and Cr were found at concentrations higher than the limit values in some of the foodstuffs. Children in Chengdu intake more metals compared to adults, with the same order of Mn > Zn > Cu > Sr > Cr > Ni > As > Cd > Pb > Co > Sb. Among all of the diverse food, rice, flour, and fish and seafood were the primary sources to intake metals for Chengdu residents. Residents in Chengdu are subjected to both carcinogenic and non-carcinogenic risks based on the calculated HI and CR values, especially for children. Finally, total daily metals intakes for both children and adults were calculated based on the current study and our previous studies, including consumption of food and drinking water and intake of outdoor and indoor dust. Dietary exposure is the predominant exposure route to metals for Chengdu residents, accounting for more than 75.8% of the total daily metals intakes for both children and adults.

Concentrations and health risk assessment of metal(loid)s in indoor dust from two typical cities of China.

Eleven trace metal(loid)s were determined in the household dust samples from Chengdu and Tianjin, China, and related human exposure and health risk to metal(loid)s via indoor dust intake were evaluated. The trace metal(loid)s were found to be highly concentrated and polluted in the indoor environment of Chengdu and Tianjin, especially for Cu, Zn, Cd, Sb, and Pb, of which the enrichment factors exceeding 5. Metal(loid) levels in the indoor dust samples exhibited no statistical differences between the two cities, with the exception of Sb, which was detected higher in the Chengdu samples. Bioaccessibilities in stomach phase of each element were estimated, Cd, Pb, and Sr exhibited higher bioaccessibility, and Sb showed the lowest bioaccessibility in both Chengdu and Tianjin. Dust ingestion was the main metal(loid) exposure pathway for Chengdu and Tianjin inhabitants, followed by dermal contact, dust inhalation accounted for less than 1 % of the total daily metal(loid) intakes and thus could be negligible. Children suffered more risk when exposure to metal(loid)s via indoor dust intake due to their higher frequency of hand to mouth activities. Risk evaluation indicated that, for most Chengdu and Tianjin inhabitants, there is little non-cancer and carcinogen risk when exposure to indoor dust. However, there is a potential non-cancer and carcinogen risk for children and adults in Chengdu, in the case of highly exposed scenario based on the current study.

Grain Size Distribution and Health Risk Assessment of Metals in Outdoor Dust in Chengdu, Southwestern China.

A total of 27 outdoor dust samples from roads, parks, and high spots were collected and analyzed to investigate the contamination of 11 metals (Cr, Mn, Co, Ni, Cu, Zn, As, Sr, Cd, Sb, and Pb) in Chengdu, China. The results showed that the samples from the high spots exhibited the highest heavy metal level compared with those from the roads and the parks, except for Ni, Cu, and Pb. The dust was classified into five grain size fractions. The mean loads of each grain size fraction of 11 determined metals displayed similar distribution, and the contribution of median size (63-125, 125-250, 250-500 µm) fractions accounted for more than 70% of overall heavy metal loads. The health risk posed by the determined metals to human via dust ingestion, dermal contact, and inhalation was investigated. Oral and respiratory bioaccessible parts of the metals in dust were extracted using simulated stomach solution and composite lung serum. The mean bioaccessibilities of 11 investigated metals in the gastric solution were much higher than those in the composite lung serum, especially Zn, Cd, and Pb. Ingestion was the most important exposure pathway with percentage greater than 70% for both children and adults. Risk evaluation results illustrated that children in Chengdu might suffer noncarcinogenic risk when exposed to outdoor dust. Given that the cancer risk values of Pb and Cr larger than 1 × 10(-4), potential carcinogenic risk might occur for Chengdu residents through outdoor dust intake.

Particle size distribution and perchlorate levels in settled dust from urban roads, parks, and roofs in Chengdu, China.

A total of 27 settled dust samples were collected from urban roads, parks, and roofs in Chengdu, China to investigate particle size distribution and perchlorate levels in different size fractions. Briefly, fine particle size fractions (<250 µm) were the dominant composition in the settled dust samples, with mean percentages of 80.2%, 69.5%, and 77.2% for the urban roads, roofs, and the parks, respectively. Perchlorate was detected in all of the size-fractionated dust samples, with concentrations ranging from 73.0 to 6160 ng g(-1), and the median perchlorate levels increased with decreasing particle size. The perchlorate level in the finest fraction (<63 µm) was significantly higher than those in the coarser fractions. To our knowledge, this is the first report on perchlorate concentrations in different particle size fractions. The calculated perchlorate loadings revealed that perchlorate was mainly associated with finer particles (<125 µm). An exposure assessment indicated that exposure to perchlorate via settled road dust intake is safe to both children and adults in Chengdu, China. However, due to perchlorate mainly existing in fine particles, there is a potential for perchlorate to transfer into surface water and the atmosphere by runoff and wind erosion or traffic emission, and this could act as an important perchlorate pollution source for the indoor environment, and merits further study.

[A forty-year study on hypertension].

Since 1959 the investigations on prevalence of hypertension and studies on the prevention and treatment of this disease have been carried out. The vascular mechanism of hypertension and the depressor effect of Chinese traditional herbs were also studied in Chinese Academy of Medical Sciences. The results revealed that: (1) The prevalence of hypertension in Chinese adults increased from 7.73% in 1979 to 11.26% in 1991, both much higher than that in 1959 (5.11%). The rate of awareness, treatment and control was only 26.3%, 12.1%, and 2.8% respectively. The risk factors of hypertension included overweight and alcohol drinking. High sodium, low potassium, low calcium, and low animal protein diet were also very important risk for elevation of blood pressure. Hypertension was the most important causal risk factor of coronary heart disease and stroke. (2) Hypertension diagnosis and staging criteria were established in 1959. Secondary hypertension was found to constitute 1.1% among community hypertensive patients. The new concept of aortitis was formed and found to be the most common cause of renal vascular hypertension. Patient education together with low dose compounds of antihypertensive drugs was implicated widely. Randomized clinical trials Syst-China, Post-stroke Antihypertensive Treatment Study, Chinese Acute Stroke Trial, and Chinese Cardiac Study 1 demonstrated benefits of treatment for hypertensive, stroke or acute myocardial infarction affordable by Chinese population at large. (3) A series of functional changes and abnormalities with evident hereditary characteristics were found in the processes of cellular Ca2+ transportation, utilization, metabolism and their modulation of the vascular smooth muscle in SHR, and SHRsp, which seem to be the principal cause of the increase in peripheral vascular resistance in hypertension. (4) Alkaloid of Rauwolfia verticilata and Ligustrazine had marked depressor effect. Flavones of Radix Pueraricae could reduce the cardiac and cerebral ischemic damage and symptoms in hypertensive patients.