When the metal release is exothermic in advance, the system temperature increases, even leading to uncontrollable levels, and the process may slip out of control.Cumene hydroperoxide (CHP) is widely used in chemical processes, mainly as an initiator for the polymerization of acrylonitrile–butadiene–styrene. It is a typical organic peroxide and an explosive substance. It is susceptible to thermal decomposition and is readily affected by contamination; moreover, it has high thermal sensitivity. The reactor tank, transit storage vessel, and pipeline used for manufacturing and transporting this substance are made of metal. Metal containers used in chemical processes can be damaged through aging, wear, erosion, and corrosion; furthermore, the containers might release metal ions. In a metal pipeline, CHP may cause incompatibility reactions because of catalyzed exothermic reactions. This paper discusses and elucidates the potential thermal hazard of a mixture of CHP and an incompatible material’s metal ions. Differential scanning calorimetry (DSC) and thermal activity monitor III (TAM III) were employed to preliminarily explore and narrate the thermal hazard at the constant temperature environment. The substance was diluted and analyzed by using a gas chromatography spectrometer (GC) and gas chromatography/mass spectrometer (GC/MS) to determine the effect of thermal cracking and metal ions of CHP. The thermokinetic parameter values obtained from the experiments are discussed; the results can be used for designing an inherently safer process. As a result, the paper finds that the most hazards are in the reaction of CHP with Fe Since the 1970s, the chemical industry has been booming in Taiwan, and it has contributed to the industrial development and economic progress of the country. However, we sporadically hear of disastrous accidents in the chemical process industry ,3. In faIn practice, cumene hydroperoxide (CHP) is an initiator for the polymerization of acrylonitrile–butadiene–styrene (ABS), which has wide applications and is ubiquitous in our daily life. CHP is organic peroxide for the following reasons: its structure has a peroxy bond (–O–O–) , it has In the past, manufacturing processes used for such substances have repeatedly caused casualties or equipment damage. An organic-peroxide-based structure has an unstable double bonding group that can readily decompose with exothermic characteristics. CHP itself is incompatible, which makes it react with acids, bases, metal ions, or other substances ,10,11.Corrosion results from the surface of metal undergoing a chemical or electrochemical reaction with the substances present in the environment, which leads to the gradual erosion of the metal interior ,13. CorrThe main objective was to conduct dynamic research to study the stability of the CHP with metal ions, that generated by corrosion phenomena. T0 for mixed metal ions of sample. However, there is no appreciable difference in ΔHd and Tmax. After adding CHP to ZnBr2, CuBr2, or FeBr2, T0 increased to 95.0, 75.0, and 74.0 °C, respectively. For these additions to ZnBr2, CuBr2, and FeBr2, the ΔHd values were 1192.0, 1100.0, and 1246.0 J/g, respectively. The parameter Tmax was stable at 160.0 ± 5.0 °C, and the relevant values are listed in In an experiment, CHP was added to three incompatible metals, and the aforementioned method was used to achieve thermal decomposition, as shown in T0), in advance of appeared from 74.0–95.0 °C, as shown in T0) of CHP is 105.0 °C. Hence, metal ions can be said to cause the reaction that affects CHP, let it T0 was in advance. It is noteworthy that, apart from the ion curves of Cu2+ and Fe2+, T0 can have advanced phenomenon, and its peak value is far from the single peak. However, ΔHd is maintained within 1150.0% ± 10% J/g, where 10% is the experimental error. The reason is that, in this test, only CHP is used to provide heat, and the metal ion does not generate heat, and therefore, the metal ion does not affect their heat of decomposition.After adding metal ions to CHP, it will let the exothermic onset temperature  for the scanning analysis at 90.0, 100.0, and 110.0 °C were 57.41, 22.88, and 9.49 h. The TMRiso values for cumene hydroperoxide (CHP) mixed with FeBr2, tested using TAM III scanning analysis at 80.0, 90.0, 100.0, and 110.0 °C were 0.34–0.43 h. Metal ions affect the reactions involving CHP by increasing the reactivity of CHP, and they change the heat mechanism markedly—especially, CHP added with incompatibility material FeBr2. The process should be performed very carefully once the system in the incompatible sections is operated in such a real hazard. CHP is affected by the addition of metals. Storage tanks, pipelines, and reactors used in the manufacture of CHP are all made of metals, and contact of the metals with CHP is inevitable.CHP was used in the isothermal evaluation by using a TAM III scanning analysis at 80.0, 90.0, 100.0, and 110.0 °C. The heat determined in a reaction kinetics analysis of substances can be used to enhance the safety of the process design. Each stage of the process may be simulated by assuming faulty conditions, and the simulation results can be used for improving the processing phase, storage, and transport. In these experiments, the TAM III thermostat obtained the relevant parameters through calculations involving the Arrhenius equation and the apparent activation energy.aE):k0 is the reaction rate constant, A is the frequency factor, aE is the apparent activation energy, R is the ideal gas constant, and T is the absolute temperature.In the thermostat tests performed in this study, 80.0 mass% CHP was considered at 80.0, 90.0, 100.0, 110.0 °C and the following equation was used to calculate the apparent activation energy  gives:aE and A are the only constant values that do not change with the temperature. The greater the temperature, the reactions are faster. Therefore, the following equation can be regarded as a set of dynamic equations:T1, T2, T3 ... are the temperatures in the absolute experiment and k1, k2, k3 ... are constants at the temperatures T1, T2, T3 ..., respectively.In the Arrhenius equation, aE value of pure CHP is 155.5 kJ/mol. Furthermore, adding a metal layer containing materials incompatible with CHP leads to a decrease in aE, and the order of aE values was ZnBr2 (146.8 kJ/mol), CuBr2 (95.6 kJ/mol), and FeBr2 (21.8 kJ/mol). The minimum energy required to kick off a chemical reaction is called apparent activation energy. When molecules collide, the kinetic energy of the molecules can be used to bend, stretch, and ultimately break bonds, leading to chemical reactions. The lowering of apparent activation energy of CHP with Fe2+ means that they are easy to start to react compared with the other ions.From the results obtained using the Arrhenius equation and apparent activation energy of the experimental group, we can derive the results, as displayed in aE of CHP, obtained in the present study, with the values obtained from the previous studies.To verify the accuracy of this experiment, 2+, Cu2+, Fe2+ as experimental factors, and finally evaluated thermal cracking products by GC/MS analysis to determine product profiles.Organic peroxides are widely used in industrial processes because of their high reactivity. Frequent disasters related to the manufacture of organic peroxides may also occur. CHP is prevailingly used for the manufacture of phenol, acetone, α-methylstyrene (AMS), and other common chemicals. To understand the corrosion of metal containers and release of metal ions during manufacturing process, we chose the trace amount of Zn2, CuBr2, and FeBr2, obtained through GC/MS analysis. Like the aforementioned atlas of pure substances, at 12.243, 16.564, and 23.337 min, the three substances—cumene, AMS, and AP—appear sequentially. Cumene is presumed to be a product of thermal volatilization. In addition, the spectrum obtained for the addition of zinc ions at 24.914 min is that of phenylethyl methyl ether. Furthermore, at 30.670 min, ethanedione, (4-methylphenyl) phenyl, appears. The spectrum of copper ions, α-cumyl alcohol (CA), appears at 24.914 min, and ethanedione, (4-methylphenyl) phenyl, evolves at 30.669 min. The spectrum of ferrous ions shows that the fourth crest is that of α-ethyl-α-methylbenzyl alcohol at 24.914 min. Finally, at 30.669 min, it displays the waveform of dicumyl peroxide (DCP).Subsequently, the three incompatible metal materials were added. 2 and FeBr2 to CHP, the heat wave peak appears twice. However, in the GC/MS experiments, within the first three substances, only CA repeated, as shown in each atlas. Furthermore, the derivatives CA and DCP were obtained by adding Cu2+ and Fe2+ in the CHP preparation process. It is predicted that this addition should generate metal ions that catalyze the formation of the two derivatives. Moreover, in the DSC experiment, the heating process in an exothermic reaction corresponds to the second thermal peak.According to individual normal curves obtained from DSC shown in 2+. The proposed mechanisms for metal-catalyzed decomposition + + H2O;C6H5C(CH3)2O]+ → [C6H5O(CH3)2C]+;[C6H5C(CH3)2OOH + [C6H5O(CH3)2C]+ → [C6H5C(CH3)2O]+ + (CH3)2CO + C6H5OH;C6H5C(CH3)2OOH → C6H5C(CH3)2O‧ + OH‧;C6H5C(CH3)2O‧ + C6H5C(CH3)2H → C6H5C(CH3)2‧ + C6H5C(CH3)2OH;C6H5C(CH3)2‧ + OH‧ → C6H5CH3C = CH2 + H2OCA comparison of the hazard posed by the mixture of CHP and incompatible metals in TAM III and reactions involving CHP contaminated with impurities show that the greatest hazard is posed by CHP mixed with Feposition  are as f2, and ZnBr2, each with a concentration of 80.0 mass%, were provided by Alfa Aesar , and FeBr2 was purchased from Acros Organics . Thermal hazard studies were conducted with 5.0 mass% CuBr2, ZnBr2, and FeBr2.CHP, CuBre measuring test crucible (Mettler ME-26732) . The test cell was sealed manually by using a kit equipped with Mettler’s differential scanning calorimetry [For determining the material energy, the sample and reference material were placed in different experimental containers. However, they were in the same heating furnace and were heated at a constant rate to measure enthalpy changes. The results were continuously recorded as an energy difference function. Dynamic scanning experiments were performed on a Mettler TA8000 system coupled with a DSC 821orimetry ,17. The 2, CuBr2, FeBr2 was added to 80.0 mass% CHP in TAM III thermostat tests at 80.0, 90, 100, and 110.0 °C, respectively.The thermal activity monitor (TAM III) and its peripherals were manufactured by Thermometric, Inc., , and the operating temperature range was from 15.0 to 150.0 °C. The sensitivity level can reach 10.0 nW, and the temperature error is ±0.01 °C. TAM III ,19,20 peGas chromatography (GC)  and gas A mass spectrometer ,23 is anTest conditions:Column: Model no: Agilent 19091S–433, −60.0 °C–325.0 °C (350.0 °C), HP −5.0 ms, Capillary: 30.0 m × 250.0 μm × 0.25 μm.Carry gas: Helium (1.0 mL/min).Injection port: Split ratio (1:1), 280.0 °C, Injection volume: 1.0 μL.Detector: MSD, 280.0 °C.With its heating mode with GC/MS, the products of qualitative analysis will use the product of each fragment peaks and GC/MS built-in database for comparison and analysis of possible species.T0 was obtained as 105.0 °C, and thermal decomposition ∆Hd was 1,086.0 J/g. The addition of metal ions did not lead to any appreciable changes in ∆Hd, but T0 increased considerably by 10.0–20.0 °C.Organic peroxides have substantial applications. In particular, they are widely used in industrial processes because of their high reactivity. In the DSC experiment, the parameter TMRiso of CHP mixed with Fe2+ at 80.0, 90.0, 100.0 and, 110.0 °C will appear during 0.34–0.43 h. TAM III test revealed that Fe2+ had the greatest influence on the extent of the CHP reaction. Iron is a major constituent of components used in manufacturing equipment. Therefore, the risk of a thermal disaster also increased substantially. The CHP manufacturing process should be performed very carefully.In the TAM III thermostat experiments, metal ions did not affect the CHP exothermic reaction. The value of GC/MS pyrolysis product analysis and CHP analysis indicated the presence of six significant peaks. Among them, the three largest peaks corresponded to cumene, AMS, and AP, which are common products in the CHP manufacturing process. In addition, they also appeared in other patterns, and they were not influenced by metal ions.2+, Cu2+, and Fe2+ patterns. Among them, CA and DCP posed a potential thermal hazard. We can predict the cause of the second peak.Investigating the GC/MS pattern of CHP containing added metal ions, we found the presence of anisole, CA, and DCP in the ZnDuring the manufacturing process, metal release may cause incompatible reactions and cause reactions to occur earlier, resulting in unforeseen events. If the monitoring system does not respond in a timely manner or adequately, thermal disasters are inevitable. Therefore, we should increase the inspection frequency of these equipment and components and periodically replace them to avoid incompatibility reactions, which can lead to disasters. If a component is corroded, it poses the risk of major accidents. Follow-up studies are required to find ways to prevent corrosions.


Suicide is the second leading cause of death among 15–29 year old youths globally, and the third most common external cause of death in China. School bullying can cause serious consequences among adolescents, including psychosocial problems, low self-esteem and post-traumatic stress disorder. Most studies examining the associations between being bullied in school and suicide are from Western, developed countries. Moreover, few studies focus on being threatened or injured with a weapon, which may cause serious body injury. Our study aimed to explore the relationship of being threatened or injured with a weapon in school with suicidal ideation and attempt among middle and high school students in Zhejiang Province, China.A cross-sectional study of 23,543 students in grades 7–12 from 442 schools was carried out through an anonymous self-administered questionnaire between April and May 2017. Multivariable logistic regression models were used to examine the relationship of being threatened or injured with a weapon in school with suicidal ideation and attempt.51.3% of participants were boys, and mean (SD) age was 15.6 (1.7) years. The overall prevalence of suicidal ideation and attempt were 16.1% (95%CI: 15.3–16.9) and 3.6% (95%CI: 3.3–4.0), respectively. Prevalence was higher among girls than boys (ideation: 19.1% vs. 13.3%. attempt: 4.4% vs. 2.9%). 13.2% of students reported being threatened or injured with a weapon in school in the past 12 months (95%CI: 11.9–14.5), higher among boys than girls (15.7% vs. 10.5%), and among rural students than urban students (13.9% vs. 11.6%). After adjustment for socio-demographic status, lifestyle factors, academic performance, self-reported health and mental health, the odds ratios for suicidal ideation and attempt among students who reported being threatened or injured with a weapon were 1.46 (1.31–1.61) and 1.68 (1.31–2.13) respectively, as compared with those who did not report being threatened or injured.Preventive measures for physical bullying in school need to be reinforced in China. Being threatened or injured with a weapon is associated with both suicidal ideation and attempt among middle and high school students. Suicide prevention should be an important component of psychological interventions for threatened or injured students. While in Western countries, mental health disorders account for more than 90% of suicides, only about half of suicides were related to mental health disorders in China [Suicide is a serious public health problem worldwide and is responsible for more than 0.8 million deaths annually. In 2012, approximately 76% of deaths due to suicide occurred in low- and middle-income countries including China . HoweverSuicidal ideation and suicide attempt are recognized as important predictors and indicators of subsequent completed suicide , 10. A mSchool bullying refers to any unwanted aggressive behaviours towards one youth by another youth or group of youths, who are not siblings or current dating partners, involving an observed or perceived power imbalance. These behaviours are repeated, or have the potential to be repeated, over time . BullyinPhysical bullying among adolescents is less normative compared to other types of bullying and therefore associated with more severe outcomes . Being tMore recently, a large number of articles have focused on the effect of being bullied in school on adolescent suicide . HoweverDetails of the study design, sampling, and procedure have been described previously . In brieSuicidal ideation was assessed by a yes/no response to the question “During the past 12 months, did you ever seriously consider attempting suicide?” Suicidal attempt was assessed through the question: “During the past 12 months, how many times did you actually attempt suicide?” Response options included: “None”, “1 time”, “2–3 times”, “4–5 times” and “6 or more times.” Suicidal attempt was defined as at least once in the past 12 months.Being threatened or injured with a weapon in school was assessed with the question: “During the past 12 months, how many times has someone threatened or injured you with a weapon such as a club or knife on school property?” Answer options included: “None”, “1 time”, “2-3times”, “4–5 times”, “6–7 times”, “8–9 times”, “10–11 times” and “12 or more times”. Students who reported one or more time were classified as being threatened or injured with a weapon.Covariate question and response options  have been reported previously , 43. SadP values < 0.05 were considered statistically significant.Statistical analyses were performed using SAS software V.9.3. A weighting factor was applied at the student-level to adjust for non-response and for the varying probabilities of selection. The weight used for estimation in this survey is given by: W = W1 × W2 × f1 × f2. W1 = the inverse of the probability of selecting the county. W2 = the inverse of the probability of selecting the classroom within the county. f1 = a student-level nonresponse adjustment factor calculated by class. f2 = a post-stratification adjustment factor calculated by grade . ContinuA total of 24,157 students from 442 schools were invited to participate. Due to missing or incomplete questionnaires or refusal to participate, 23,543 eligible students (response rate 97.5%) were included in the final analyses. 12,068 (51.3%) were boys, and mean age was 15.6 years. 12,207 (51.9%) students were from middle schools, 6477 (27.5%) were from academic high schools, and 4859 (20.6%) came from vocational high schools.P > 0.05).Compared with those who reported not being threatened or injured, students who reported being threatened or injured with a weapon in school were more likely to be boys (61.8% vs. 50.3%) (Table P < 0.0001). There was no statistically significant difference in the prevalence of suicidal ideation between urban and rural areas (16.5% vs. 15.9%) (P = 0.43). The prevalence among students aged ≤13 years, 14–15 years and ≥ 16 years was 15.9% (95%CI: 14.4–17.4), 17.9% (95%CI: 16.7–19.1) and 14.8% (95%CI: 13.7–15.9), respectively. The corresponding prevalence among students attending middle school, academic high school and vocational high school was 16.9% (95%CI: 15.8–18.0), 16.6% (95%CI: 15.3–17.9) and 13.7% (95%CI: 11.9–15.5), respectively (P = 0.0048).The overall prevalence of suicidal ideation was 16.1% (95%CI: 15.3–16.9) Table , higher P < .0001). No regional difference in suicidal attempt was found  (P = 0.50). The prevalence of suicidal attempt among students aged ≤13 years, 14–15 years and ≥ 16 years was 4.1% (95%CI: 3.5–4.8), 4.4% (95%CI: 3.9–4.9) and 2.8% (95%CI: 2.3–3.2), respectively. The corresponding figures for students attending middle school, academic high school and vocational high school were 4.4% (95%CI: 3.9–4.9), 2.4% (95%CI: 1.9–2.8) and 3.3% (95%CI: 2.6–4.1), respectively (P < .0001).The overall prevalence of suicide attempt was 3.6% (95%CI: 3.3–4.0) Table , higher P < .0001). The prevalence of being threatened or injured with a weapon increased with the decreasing age (P < .0001). There was no regional difference in the prevalence of being threatened or injured with a weapon in school among boys  (P = 0.18). However, rural girls had a higher prevalence than their urban counterparts (11.3% vs. 8.6%) (P = 0.02).13.2% (95%CI: 11.9–14.5) of students reported being threatened or injured with a weapon in school in the past 12 months , respectively, among students who reported being threatened or injured with a weapon in school. After further adjustment for parental education level and marital status, lifestyle factors, academic performance, and health status, the corresponding odds ratios were attenuated to 1.89 (1.72–2.09) and 2.16 (1.72–2.72), respectively . After additional adjustment for mental health . These associations were further markedly attenuated, with corresponding odds ratios of 1.46 (1.31–1.61) and 1.68 (1.31–2.13), respectively . No sex-difference was observed in the associations of being threatened or injured with a weapon and suicidal ideation (P = 0.61) or attempt (P = 0.07).After adjusting for age, sex, region, and type of school Table , compareIn this provincially representative study of middle and high school students in Zhejiang, China, we examined the prevalence of being threatened or injured with a weapon in school and suicidal ideation and attempt. We identified and quantified the relationship of being threatened or injured with a weapon in school and both suicidal ideation and attempt.Suicidal ideation involves a hierarchy of feelings from the thoughts of “Life is not worth living” to more serious articulation of a thought out plan. Suicidal attempt is defined as self-inflicted, potentially injurious behaviours with a nonfatal outcome, for which there was evidence of intent to die . In the In the present study, 13.2% of students reported being threatened or injured with a weapon in school during the past 12 months, more than twice the reported prevalence in the USA (6.0%) . This suA 2013 meta-analysis of 41 studies found that being bullied was positively associated with suicidal ideation  and suicide attempt among adolescents  . HoweverThere are a number of important implications of the findings of our study for prevention of adolescent suicide. First, although the Chinese government introduced a strategy for strengthening the comprehensive management of bullying, and launched an anti-bullying campaign in primary and middle schools in 2017, prevention of school bullying needs to be further reinforced in China. Students should be taught how to deal with school bullying, including being threatened or injured by others with a weapon. Second, students who report being threatened or injured with a weapon should be identified as being at-risk, and suicide prevention should be an important component of psychological support and interventions provided to these students.The strengths of the current study include the provincially representative sample, a high response rate and a standardized procedure. The study questionnaire collates data on a range of socio-demographic, behavioral and mental health factors, which allowed us to better control confounding. Our study, however, had several limitations. First, the cross-sectional design limits establishment of the temporal relationship of being threatened or injured with a weapon in school with suicidal ideation and attempt, and it is not possible to establish the causality of the relationship. Second, all data were self-reported; some information might not be accurately reported due to recall or social desirability biases. Physical bullying in school is relatively more common and familiar to teachers and parents. Non-physical bullying  is less visible and more easily neglected. In addition, with the rapid development of Internet and electronic technology, children and adolescents are more likely to be exposed to bullying in an electronic context . Hence, longitudinal studies are needed to explore the associations of non-physical and physical bullying with suicidal ideation and attempt.Despite its limitations, our study clearly described the associations of being threatened or injured with a weapon in school and suicidal ideation and attempt among middle and high school students in Zhejiang China. We found that being threatened or injured with a weapon in school was common, and was strongly associated with both suicidal ideation and attempt. Preventive measures for physical bullying among middle and high school students need to be reinforced in Zhejiang China, and efforts to prevent adolescent suicide may need to address school bullying. Suicide prevention should be an important component of psychological intervention for threatened or injured students.


The results indicate that the magnetic susceptibilities of the bus stop dusts were higher than those in the local soils. Cd, As, Ni, and Pb contents ranged from 0.4 to 3.1, 7.1 to 16.3, 12.7 to 151.3, and 20.1 to 96.2 mg kg−1, with average values of 1.2, 10.1, 22.2, and 44.9 mg kg−1, while Hg content ranged from 4.5 to 1357.7 µg kg−1 with an average of 214.0 µg kg−1. The mean contents of Cd, As, Hg, Ni, and Pb were 12.0, 0.8, 10.0, 0.6, and 2.4 times the local soil background value, respectively. Cd, Hg and Pb in approximately 96%, 62% and 19% of the bus stop dusts were above the “moderately polluted” level in terms of Igeo. As and Ni were defined as “practically unpolluted” in all of the bus stop dusts. The NSPI values of all of the bus stop dust samples exceeded 3, which reveals overall serious contamination of harmful elements.To appraise the content and pollution level of cadmium (Cd), arsenic (As), mercury (Hg), nickel (Ni) and lead (Pb) in bus stop dusts, representative samples (n = 53) were collected from the city of Qingyang in Gansu province, NW China. The Cd, As, Hg, Ni, and Pb contents and physicochemical properties  of the bus stop dusts were investigated. Pollution levels were evaluated by the Nemero synthesis pollution index (NSPI) and geoaccumulation index (I Transportation, biomass burning, coal combustion in power plants, refining, waste incineration, and fossil fuel combustion are the main sources of these elements4. Fossil fuels , which contain these toxic elements, were combusted in large quantities during the industrialization and urbanization of China and are still the main source of energy in the city of Qingyang in eastern Gansu province. The contents of Cd, As, Hg, Ni and Pb in the coal from North China are 0.1–3, 0.4–10, 0.2–0.5, 1–60, and 5–60 µg g−1, respectively5. The average contents of Cd, As, Hg, Ni and Pb in coal consumed in Gansu province are 0.1, 3.7, 0.2, 16.3, and 7.5 µg g−1, respectively6. Cd, As, Hg, Ni, Pb and other toxic elements contained in fossil fuels are discharged into the environment during combustion7. In 2012, the emissions of Cd, As, Hg, Ni and Pb from anthropogenic sources in China were approximately 527, 2529, 695, 3396, and 14398 t, respectively6. In 2013, the emissions of Cd, As, Hg, Ni and Pb from open municipal solid waste burning alone were estimated to be approximately 10, 132, 21, 44, and 97 in all of China and 0.2, 2.2, 0.4, 0.7, and 1.7 t in Gansu province, respectively8. Cd, As, Hg, Ni and Pb dispersed into the atmosphere and deposited as dusts in the water and soil through dry and wet deposition processes. The dust sources have an adverse effect on the atmosphere, water, soil, and human health.The high contents of cadmium (Cd), arsenic (As), mercury (Hg), nickel (Ni) and lead (Pb) in bus stop dusts pose a potential threat to commuters and pedestrians. Cd, As, Hg, Ni and Pb persist in the environment, occurring in both inorganic and organic forms in nature, and are toxic to human health, flora and fauna9. Street dusts can enter the human body by inhalation, ingestion and skin contact and may cause various health problems11. Numerous studies have been carried out on manganese, zinc, chrome and copper in dusts17. However, information on Cd, As, Hg, Ni and Pb in street dusts in moderate- or small-sized cities in Northwest China is very limited. Cd, As, Hg, Ni and Pb can affect the liver, skin, haematopoietic system, cardiovascular system, respiratory tract and gastrointestinal tract; represent a risk for bladder, lung, kidney, liver and skin cancer; and lead to black foot disease and Minamata disease20.The composition and quantities of toxic elements in street dusts are quality indicators of urban environmentsgeo and NSPI to classify the contamination levels of Cd, As, Hg, Ni and Pb in different bus stop dusts. These results could be used to establish layouts and quantities of bus stops and formulate appropriate environmental management and planning policies according to bus stop pollution, population and traffic volume, production and living conditions by the municipal department. Our government and authorities could reasonably plan locations and quantities of bus stops, bus routes, etc.; adopt mitigation policies; and control measures to better maintain the “blue sky plan” in the Qingyang region.Since the development of modern cities, urban buses have become the main means of transportation for citizens due to their convenience and act as an “artery” for population mobility. However, research into the impact of pollution at bus stops on environmental and public health risks is rarely reported. The purpose of this study is to analyse the content of Cd, As, Hg, Ni and Pb in bus stop dusts from Qingyang and apply I21. The low-frequency magnetic susceptibility (χLF) indicates the relative content of magnetic minerals22 and varied from 52 × 10−8 to 792 × 10−8 m3 kg−1, with an average of 213 × 10−8 m3 kg−1. These values are lower than the magnetic susceptibility of street dusts in Lanzhou (450 × 10−8 m3 kg−1)23, Xi’an (487 × 10−8 m3 kg−1)24, Beijing Olympic Park (243 × 10−8 m3 kg−1)25, Ezhou (4150 × 10−8 m3 kg−1), Zhuzhou (663 × 10−8 m3 kg−1), Hezhang (527 × 10−8 m3 kg−1)26, and Loudi (880 × 10−8 m3 kg−1)27 and higher than the magnetic susceptibility of street dusts in Wuhan (132 × 10−8 m3 kg−1)28, Seoul (62 × 10−8 m3 kg−1)29, and Plovdiv (193 × 10−8 m3 kg−1)30. The frequency-dependent magnetic susceptibility (χFD) indicates the relative content of superparamagnetic minerals23 and varied from 0.1% to 13.5%, with an average of 3.0%. These values are lower than the χFD of street dusts in Hezhang (4.4%) and Zhuzhou (3.3%)26 and higher than the χFD of street dusts in Wuhan (1.0%)28, Beijing Olympic Park (1.6%)25, Lanzhou (2.2%)23, and Ezhou (2.9%)26.Table FD and χLF of road dusts can reflect the effect of anthropogenic activities23. Table LF and lower χFD. The dust samples collected from bus stops in areas with a high volume of vehicles and dense population, such as bus stations, motor vehicle maintenance service areas, schools, and parks, had larger χLF values, likely due to the combined result of a high density of vehicles, oil varnish, lubricating oil, and other production and operating activities near these bus stops. The dust samples collected from bus stops in areas with a low density of vehicles, such as residential areas and hotels, had small χLF values, likely due to the low volume of vehicles and few other sources and operating activities near these bus stops.The values of χThe organic matter (OM) content in the bus stop dust samples ranged from 2.8% to 18.8%, with a mean value of 11.5%. The dust samples collected from bus stops near schools, parks, hotels and residential areas had a higher OM content.31. Airborne PM 2.5 and PM 10 can seriously affect human health, as they can directly enter the respiratory system through inhalation. In addition, the sampling method used can result in a loss of particles, thus altering the original particle size fractionation. Therefore, the mean values indicate that approximately one-fourth of bus stop dusts could potentially enter the respiratory system (PM ≤ 10 comprises 24.1%), and resuspended PM (PM ≤ 50) in the atmosphere may comprise more than 63.0% of all bus stop dusts. Therefore, the Qingyang bus stop dusts have a large potential to degrade atmospheric quality and cause harm to human health.The particle size analysis results indicate that the proportion of PM >50, PM 10–50, PM 2.5–10 and PM <2.5 in the bus stop dusts from Qingyang ranged from 6.5% to 62.4%, 8.0% to 58.7%, 5.0% to 35.8% and 2.1% to 11.3%, with averages of 37.0%, 38.9%, 18.0% and 6.1%, respectively. Particulate matter with diameters below 100 µm (PM 100) can be transported by suspension, while particulate matter with diameters below 10 µm (PM 10) can remain airborne over a long distance and for a long time−1)  and the greenbelt (sample 18) . The Cd contents in all of the bus stop dust samples are 4.0 to 31.0 times the Cd content in GS . Most of the dust samples from bus stop near hospitals  and shopping centres  , schools , and hotels (samples 39 and 49) . Most of the dust samples from bus stops near bus stations (sample 37), residential areas (sample 36), vehicle maintenance service sites (sample 1) and shopping centres (sample 42) had higher Pb contents  and Ni (35.2 mg kg−1) in GS. These results show that most bus stop dust samples contain small amounts of As and Ni.Table Table geo results for Cd, As, Hg, Ni and Pb in the Qingyang bus stop dusts. The values of Igeo for Cd, As, Hg, Ni and Pb ranged from 1.3 to 4.2, −1.4 to −0.2, −2.4 to 5.8, −2.1 to 1.5 and −0.5 to 1.8, with averages of 2.8, −0.9, 2.3, −1.4 and 0.6, respectively. Figure geo and approximately 64% of the Igeo values for Cd were between 2 and 3, indicating that the bus stop dusts were “moderately to strongly” polluted with Cd, while approximately 32% of the Igeo values for Cd were over 3, revealing that the bus stop dusts were “strongly” polluted. The average Igeo and approximately 26% of the Igeo values for Hg were between 2 and 3, indicating that the bus stop dusts were “moderately to strongly” polluted with Hg, while approximately 36% of the Igeo values for Hg were over 3, revealing that the bus stop dusts were “strongly” polluted. The average Igeo and approximately 77% of the Igeo values for Pb were between 0 and 1, indicating that the bus stop dusts were “slightly to moderately” polluted with Pb, while approximately 19% of the Igeo values for Pb were between 1 and 2, revealing that the bus stop dusts were “moderately” polluted.Table The results of the Nemero synthesis pollution index (NSPI) analysis of Cd, As, Hg, Ni and Pb in the bus stop dust samples are shown in Fig. FD and higher χLF, which reflect the frequency of anthropogenic activities. Resuspended particulate matter comprises over 63% of the bus stop dusts, and about one-fourth of the bus stop dusts have the potential to directly enter the respiratory system. The geoaccumulation index indicates that approximately 96%, 93% and 19% of the bus stop dusts are more than “moderately polluted” by Cd, Hg and Pb, respectively, while most of the bus stop dusts could not be classified as polluted by As and Ni. The prime anthropogenic sources of Cd, Hg and Pb in bus stop dusts from Qingyang are likely to be the high vehicle density, lubricating oil, welding activities, mercury thermometers and sphygmomanometers, large population and vehicle flow, building and infrastructure construction development, decoration, and so on. The NSPI values reveal overall serious contamination by toxic elements. Therefore, our government and agencies could ameliorate pollution at bus stops in Qingyang by considering vehicle and population flow, production and living conditions, and authorities should plan reasonable locations and quantities of bus stops, bus routes, etc. and adopt mitigation policies and control measures to better maintain the “blue sky plan” in the Qingyang region.Cd, As, Hg, Ni and Pb pollution and the physicochemical properties of bus stop dusts from Qingyang were studied. The pH of the bus stop dusts is slightly higher than the background value of Gansu soil. Most bus stop dusts have the characteristics of lower χ2, with a total population of approximately 325.2 thousand in 2015. The city has a characteristic semi-arid temperate continental monsoon climate with the following annual averages: evaporation capacity of 1613.1 mm, precipitation capacity of 465.7 mm, temperature of 9.6 °C, wind velocity of 2.7 m s−1, and sunlight of 2490.4 h. The moisture content in the local soil is 11.7%, and the electrical conductivity is 92.4 s cm−1. The texture analysis indicates that the soil is composed of 8.1% clayey soil, 39.5% silty sand and 52.4% fine sand21.The city of Qingyang (35°15′–37°10′N and 106°20′–108′45′E) is situated in the Loess Plateau region in eastern Gansu province  and low-frequency signals  were measured by a Bartington magnetic susceptibility meter . The frequency-dependent susceptibility (χFD) was calculated as χFD = (χLF − χHF)/χLF × 10048. The pH was measured in a 2:1 mixture of ultrapure water and sample34. In addition, the particulate matter (PM) was measured by a laser analyser . The organic matter (OM) was measured by the loss-on-ignition method of Xie33. Samples were oven-dried at 105 °C, weighed and then ignited in a muffle furnace at 400 °C for 4 h before being re-weighed to determine the OM loss.High-frequency  in a polytetrafluoroethylene (PTFE) pipe. The PTFE pipe was covered and placed in a microwave digestion system at 180 °C for 1 h. The contents of As and Hg in the bus stop dust samples were determined by double channel atomic fluorescence spectrometry35. The content of Cd in the bus stop dust samples was determined by graphite furnace atomic absorption spectroscopy36. The total concentrations of Pb and Ni were determined by flame atomic absorption spectroscopy 35.Each sample (0.5 g) was digested in a mixture of HNO3 (8%) for at least 20 h and rinsed three times with tap water, secondary deionized water and ultrapure water, respectively. Quality control (QC) samples were a standard soil substance (GBW07406-GSS-6) and standard reagents  and were injected after every 18 samples to assess the stability of the instrument. The variations in the metal concentrations of the QCs were <13%. Reagent blanks were also included in each batch of analyses to check for any cross contamination of the different digested samples. The precision of the analytical procedures was expressed as the relative standard deviation (RSD), which ranged from 3% to 8% and was calculated from the standard deviation divided by the average value. Calibration curves were prepared separately for each element using different concentrations  of the standard solutions. The instrument was set to zero concentration for all samples using a reagent blank. Each determination was based on the mean value of three replicate measurements, the detection limits of Cd, As, Hg, Ni and Pb are 0.1 µg kg−1, 13 µg kg−1, 3 µg kg−1, 0.3 µg kg−1, 0.06 mg kg−1, respectively. The statistics were computed using Origin 8.0 software and Microsoft Office Excel 2016.Analytical-grade reagents and chemicals, purchased from Tianjin Kemiou Chemical Reagent Co., Ltd. (China), were used for sample analysis. All solutions were prepared using ultrapure water from a Milli-Q System . All glassware and plastics were soaked in HNO40. The geoaccumulation index (Igeo) and Nemero synthesis pollution index (NSPI) are used frequently as environment evaluation methods. Therefore, assessment of the element pollution levels in the bus stop dusts used both Igeo and NSPI.There are numerous calculation methods for quantifying the degree of harmful element enrichment or contamination in dusts and soilsn is the background content of the element n in the Gansu soil sample and Cn is the measured content of element n in the bus stop dusts. When Igeo ≤ 0, the bus stop dusts are non-polluted; when 0 < Igeo ≤ 1, the bus stop dusts are slightly to moderately polluted; when 1 < Igeo ≤ 2, the bus stop dusts are moderately polluted; when 2 < Igeo ≤ 3, the bus stop dusts are moderately to heavily polluted; when 3 < Igeo ≤ 4, the bus stop dusts are heavily polluted; when 4 < Igeo ≤ 5, the bus stop dusts are heavily to extremely polluted; and when 5 < Igeo ≤ 10, the bus stop dusts are extremely polluted42.The following equation was used to calculate the results:max and PImean are the max value and average value of the pollution index (PI) of five harmful elements in every sample. The PI is the ratio of the content of a certain element in the bus stop dusts to the corresponding background value of the element in Gansu soil43. When NSPI < 0.7, the bus stop dusts are non-polluted; when 0.7 < NSPI < 1, the bus stop dusts are nearly polluted; when 1 < NSPI < 2, the bus stop dusts are slightly polluted; when 2 < NSPI < 3, the bus stop dusts are moderately polluted; and when NSPI > 3, the bus stop dusts are heavily polluted40.The NSPI can be obtained by the following equation:


They utilized it as monomer for the synthesis of non-isocyanate polyurethanes (NIPUs). Here, we present a significantly more sustainable procedure for the carbonate formation regarding solvent, carbonyl source, reaction temperature, reaction time, reduced pressure during the reaction, simplicity of the workup as well as recycling of reagents. Catalysed by triazabicyclodecene (TBD), dimethyl carbonate as solvent as well as reagent leads to selective product formation and facile product separation by filtration. After addition of new starting materials, the mixture of catalyst and DMC was reused up to 8 times without loss of catalytic activity.Recently, R. Mülhaupt  Several methods are available for the synthesis of non-isocyanate polyurethanes (NIPU)3. A first step towards more sustainable polyurethanes was the synthesis of isocyanates using the Curtius rearrangement to avoid the use of phosgene, even though it has to be viewed critically, as it proceeds via toxic acyl azides4. Sustainable polyurethanes with backbones similar to the petroleum-based ones can be produced by transurethanization of carbamates7. Moreover, a sustainable synthesis of bio-based carbamates was performed by Lossen rearrangement of hydroxamic acids, which are activated in situ by dialkyl carbonates in the presence of catalytic amounts of tertiary amine bases (0.1–0.4 eq.)8. Still, NIPU synthesis from bis-cyclic carbonates (bCC) and diamines is arguably the most promising approach9, as the obtained molecular weights are typically higher than with the other approaches. The first route for the synthesis of five-membered carbonates is on the example of ethylene carbonate and involves phosgenation of ethylene glycol by phosgene or triphosgene, which is due to obvious reasons not suitable for a sustainable synthesis, as the original purpose of NIPUs is to avoid phosgene10. More sustainable approaches are typically based on the insertion of CO2 into epoxides, which are synthesized from renewable resources containing double bonds or from 1,2-diols.Polyurethanes are among the most widely used polymers in industry – in particular for specialty applications in furniture, construction, electronics, automotive and packaging. Typically, polyurethanes are obtained by polymerization of polyols with diisocyanates as co-monomer. However, the use of isocyanates is not considered sustainablee.g. glycerol carbonate, the direct synthesis from the 1,2-diol and supercritical CO2 (SCCO2) has been established11, although the reaction is currently still not feasible on an industrial scale due to low conversions (32%)2. Another method utilizing supercritical CO2 was reported by M. Aresta et al. first obtaining the ketal of cyclohexanone and 1,2-ethanediol followed by transition-metal complex catalyzed carbonate formation in SCCO2 or with CO2 in organic solvents12. Other methods involving pressurized CO2 and 1,2-diols are performed in acetonitrile at 170 °C and catalyzed by metallic acetates13, by electrosynthesis14, or with diazabicycloundecen in dibromomethane15. The carbonate interchange reaction between 1,2-diols and ethylene carbonate or linear carbonates 18 is more common and also used for the herein reported approach. A different type of transesterification can be achieved with urea and ZnO or various other solid catalysts releasing ammonia in the process19. Very interesting is the oxidative carbonylation with carbon monoxide using, for instance, palladium-based catalysts21, which is also the established synthesis of most linear carbonates. Alternatively to a two-step procedure (first synthesizing the epoxide and insertion of CO2 afterwards), olefins can be directly converted by oxidative carboxylation with a catalytic system of MoO2(acac)2, a quaternary ammonium salt and tert-butyl hydroperoxide as an oxidant in an one-pot multistep process22. Other methods available for this oxidative carboxylation were reviewed in 2011 by J. Sun and coworkers23. Methods involving other starting materials are of minor importance, e.g. from halohydrins23, substituted propargyl alcohols25, halogenated carbonates26, or linear oligo-carbonates29. Six- or seven- membered cyclic carbonates can be obtained from 1,3-diols or 1,4 diols by similar means as five-membered cyclic carbonates. While they provide a better reactivity due to a lower thermodynamic stability 9, the synthesis of six- or seven- membered carbonates is significantly more difficult for exactly the same reason30. Still, K. Tomishige et al. reported a particularly efficient catalyst (CeO2) with high yields for six- membered carbonates (62–99%) from CO2 and diols in 201431. However, a high excess (10 times) of 2-cyanopyridine was needed to remove water from the reaction mixture and obtain a high selectivity.For some carbonates, such as bis(carbonate) (EBC) was already synthesized in 1960 from butadiene and patented by Union Carbide Whelan for polyurethane synthesis32, however it can also be synthesized in a more sustainable fashion by transesterification from the renewable sugar substitute erythritol. Erythritol is a narurally occuring sugar alcohol, which is present in many fruits and vegetables and used as a bulk sweetener. It is industrially produced by a fermentation process with the naturally occurring yeast Moniliella pollinis from aqueous solutions of glucose or sucrose33. Starting from erythritol, several routes to EBC are known. T. Griesser et al. achieved high yields up to 88%, however the phosgene based 1,1′-carbonyldiimidazole was used as carbonyl source34. Until now, the carboxylation of erythritol with dimethyl carbonate was claimed to be unfeasible due to low yields35, since in 2012 G. Rokicki et al. reported it as side reaction of an intramolecular etherification with only 10% yield36. They utilized an excess of dimethyl carbonate (DMC) at 70 °C and 226 mbar with K2CO3 as catalyst, only obtaining the desired EBC in 5% yield. In 2017, based on these results, R. Mülhaupt et al. used diphenyl carbonate (DPC) instead of DMC at 120 °C and 30 mbar in DMSO with Zn(OAc)2 as catalyst, thus avoiding the intramolecular etherification and obtaining EBC in 80–90% yield35. Additionally, the same method was later applied to sorbitol in order to obtain sorbitol tricarbonate37. Even though the yield is good, the synthesis lacks in sustainability regarding several aspects. DPC is far less sustainable than DMC. Aside from the classic unsustainable phosgenation of alcohols, both can be produced by oxidative carbonylation from the respective alcohol and carbon monoxide38. However, while phenol is typically produced in the cumene process from fossil benzene and propene39, methanol can be obtained via various different routes from renewables (e.g. from bio-derived synthesis gas or methane)40. Moreover, the process of the oxidative carbonylation of methanol is more efficient and involves cheaper catalysts  and auxiliaries than the one of phenol. Industrially, the oxidative carbonylation of phenol is barely realized, while for methanol this route is established38. Here, we report on an efficient and sustainable synthesis of EBC based on DMC , but as it can be recovered and reused, the overall price might not be much higher. The workup consisted of a simple filtration compared to a recrystallization, which requires additional solvent and thus results in additional waste, as it cannot be reused or recycled. When testing the developed reactions conditions with sorbitol, another important renewable alditol that could yield a useful tricarbonate for NIPU synthesis, unfortunately, significantly lower conversion as well as selectivity was observed. Thus, the development of specific conditions for each alditol seems necessary. For instance, L. Avérous et al. reported a yield of up to 50% of -6--2,4,7trioxa-3-oxy-bicyclo[3.3.0]octane, a bCC from D-sorbitol obtained after intramolecular etherification, using 5 mol% TBD in DMC at 75 °C and a reaction time of 16 hours43.DMSO, the solvent for the literature procedure, achieves a total rating of “some issues” in GSK’s solvent sustainability guide, while DMC is among the few solvents with “few issuessee Fig. . By usinbis(carbonate) – a valuable monomer for NIPU synthesis – was obtained from erythritol featuring mild reaction conditions and a facile workup in 90% yield. The reusability of catalyst and reaction mixture was demonstrated up to 8 times, while the catalyst showed no sign of decrease in activity. In principle, the developed procedure might enable a one-pot polymerization by directly adding the diamine to the reaction mixture due to its high selectivity as well as the presence of a TBD as a suitable catalyst, but keeping the catalyst in the monomer synthesis cycle even longer seems more practical and more sustainable.In conclusion, erythritol m/z at rate of 1.0 scans s−1. The oven temperature program was: initial temperature 95 °C, hold for 1 min, ramp at 15 °C min−1 to 220 °C, hold for 4 min, ramp at 15 °C min−1 to 300 °C, hold for 2 min. The injector transfer line temperature was set to 250 °C. Measurements were performed in the split – split mode (split ratio 50:1) using helium as carrier gas (flow rate 1.0 mL min−1).GC-MS  measurements were performed on the following system: a Varian 431 GC instrument with a capillary column FactorFour VF-5 ms (30 m × 0.25 mm × 0.25 mm) and a Varian 210 ion trap mass detector. Scans were performed from 40 to 650 1H- and at 75 MHz for 13C-measurements) were recorded on a Bruker AVANCE DPX spectrometer operating at 300 K. For all NMR-spectra the residual non-deuterated solvent (1H NMR) or CDCl3 (13C NMR) signal was used as internal standard. Infrared spectra (IR) were recorded on a Bruker Alpha-p instrument in a frequency range from 3998 to 374 cm−1 applying ATR technology.NMR spectra (300 MHz for ESI-MS spectra were recorded on a Q Exactive (Orbitrap) mass spectrometer  equipped with a HESI II probe to record high resolution electrospray ionization – MS (ESI-MS). Calibration was carried out in the m/z range of 74–1.822 using premixed calibration solutions (Thermo Fisher Scientific). A constant spray voltage of 4.7 kV and a dimensionless sheath gas of 5 were employed. The S-lens RF level was set to 62.0, while the capillary temperature was set to 250 °C.In a 100 mL flask, erythritol  and TBD  were dispersed in 41 mL DMC and heated to 60 °C for 40 min at the rotary evaporator at 314 mbar. The crystalline erythritol dissolved completely after 30 min and after 35 min, a white precipitate was formed. The product was filtered off after the mixture was cooled down to room temperature and washed with dimethyl carbonate yielding a white powder .1H-NMR : δ (ppm) = 5.25–5.07 , 4.69–4.52 , diastereotopic signals), 4.47–4.31 ; 13C-NMR : δ (ppm) = 158.2 (CO), 74.9 (CH), 64.7 (–CH2-); HRMS (ESI) of C6H6O6 [M + H]+m/z calc. 175.0237, found 175.0234; IR (ATR): ν = 1803.7, 1779.0, 1545.5, 1476.1, 1381.2, 1299.6, 1206.5, 1144.6, 1067.0, 1030.9, 978.1, 896.2, 771.8, 739.1, 719.1, 532.1, 384.9 cm−1.In a 100 mL flask, erythritol  and TBD  were dispersed in 41 mL DMC and heated to 60 °C for 40 min at the rotary evaporator at 314 mbar. The crystalline erythritol dissolved completely after 30 min and after 35 min, a white precipitate was formed. The product was filtered off after the mixture was cooled down to room temperature and washed with 15 mL dimethyl carbonate, which was combined with the mother liquor, to obtain the product  and again 41 mL volume of a mixture containing the catalyst and non-precipitated product. Another batch of erythritol  was dispersed in the mother liquor and the procedure was repeated for 7 additional times yielding on average 90 ± 1% (procedure carried out in triplicate) of the product.


Hypertension is a highly prevalent disease and the leading cause of chronic kidney disease (CKD). Metabolic syndrome could also be the risk factor for CKD. We sought to study the association between metabolic syndrome components and the prevalence of CKD in patients with hypertension.We carried out a multi-center cross-sectional study from Apr. 2017- Apr. 2018 in 15 cities in China.p < 0.01), elevated fasting blood glucose (FBG)  and hypertension grades  were associated with the prevalence of CKD. In sub-group analysis, elevated TG remained strongly associated with CKD in both diabetes  and non-diabetes . In sub-group analysis of hypertension grades, there was also a graded trend between elevated TG and CKD from controlled blood pressure (BP) to hypertension grade 2 .A total of 2484 patients with hypertension were enrolled. Among them, 56% were male and the average age was 65.12 ± 12.71 years. The systolic BP/diastolic BP was 142 ± 18/83 ± 12 mmHg. Metabolic syndrome components turned out to be highly prevalent in patients with hypertension, ranging from 40 to 58%. The prevalence of chronic kidney disease reached 22.0%. Multi-variate logistic analysis revealed that elevated triglyceride (TG) (OR = 1.81, 95% CI 1.28–2.57, Elevated TG, elevated FBG and hypertension grades were significantly associated with the prevalence of CKD in patients with hypertension. Particularly, elevated TG was strongly associated with CKD, independent of diabetes and hypertension grades. Hypertension is highly prevalent worldwide. It has been reported that 31.1% (95%CI 30.0–32.2%) of the world’s adults have hypertension. However, the awareness, treatment and control rate are far from satisfying. Data from different countries showed that the awareness of hypertension ranged from 37.9–67%, treatment rate ranged from 29.0–55.6% and the control rate ranged only from 7.7–28.4% [We carried out a multi-center cross-sectional study from Apr. 2017-Apr. 2018 in the main 15 cities in China. Participants were enrolled from hypertension clinic using convenience sampling. Inclusion criteria were patients with primary hypertension, aged over 18 years, who could provide informed consents. Patients were excluded if they had secondary hypertension, genetic renal diseases, active cancer or recognition disorder. Each patient completed a questionnaire collecting information on demographic characteristics, medical history, lifestyle features, current medical treatment, physical examination and lab tests.According to American Heart Association, the diagnosis of metabolic syndrome includes any 3 of the 5 criteria: 1) Elevated waist circumference > =90 cm in men and > =80 cm in women (for Asian); 2) Elevated triglycerides (TG) > =1.7 mmol/L; 3) Reduced HDL-C < 1.03 mmol/L in men or < 1.3 mmol/L in women; 4) Elevated blood pressure > =130/85 mmHg or on antihypertensive drug treatment with a history of hypertension; 5) Elevated fasting blood glucose (FBG) > =5.6 mmol/L or on drug treatment for elevated glucose [As is listed above, in our study, elevated waist circumference was defined as waist circumference > =90 cm in men and > =80 cm in women; elevated TG was defined as TG > =1.7 mmol/L; reduced HDL-C was defined as HDL-C < 1.03 mmol/L in men or < 1.3 mmol/L in women; elevated FBG was defined as FBG > =5.6 mmol/L or on drug treatment for elevated glucose.Hypertension grades was defined according to 2013 ESC/ESH Guidelines for the management of arterial hypertension  . Contro2, while eGFR was calculated using Modification of Diet in Renal Disease Study equation [CKD was defined as estimated glomerular filtration rate (eGFR) < 60 ml/min/1.73mBaseline characteristics were analyzed and compared in patients with and without CKD. Either two-side Student’s t-test or Wilcoxon rank sum test were used for continuous variables. Chi-square test was used for categorical variables. A forward stepwise method was used to reach the logistic regression model. Odds Ratios (OR) and 95% confidence intervals from logistic regression were used to estimate the association between metabolic syndrome components and CKD. Sub-group analysis was conducted according to concomitant diabetes and hypertension grades. P < =0.05 was considered statistically significant. STATA 13.0 was used for statistical analysis.In the 2484 patients with hypertension, the average age was 65.12 ± 12.71 years. Male accounted for 55.9% of the total participants. The average BP was 142.30 ± 17.68 / 83.01 ± 11.93 mmHg. The distribution of hypertension grade was 41.6, 36.9, 14.9 and 6.7% separately. Totally 90.7% of them were taking antihypertensive therapy.Metabolic syndrome turned out to be highly prevalent. Patients with elevated TG, reduced HDL-C, elevated FBG and elevated waist circumference were 46.9, 40.1, 58.3 and 47.5% separately. Besides, 24.7% of them had diabetes and 24.2% of them were taking antidiabetic therapy. Additionally, 60.3% were using statins and 0.8% were using fibrates.The prevalence of chronic kidney disease was also high, reaching 22.0% of the total participants Table .Table 1p < 0.01). Among them, 57.9% were male in non-CKD group and 49.0% were male in CKD group (p < 0.01). No difference was found in the distribution of hypertension grades (p = 0.16) and antihypertensive therapy (p = 0.99) between the two groups. The ratio of elevated TG, reduced HDL and elevated FBG were 45.2,39.5 and 56.6% in non-CKD group compared to 54.1% (p = 0.01), 42.5% (p = 0.35) and 66.1% (p = 0.01) in CKD group. The elevated waist circumference was 45.7% in the non-CKD group compared to 54.0% in the CKD group (p < 0.01). The prevalence of DM was 23.7% in non-CKD group compared to 28.3% in CKD group (p = 0.03). Totally 22.8% of the patients in non-CKD group were taking antidiabetic therapy compared to 29.2% in CKD group (p < 0.01). Additionally, 58.2% patients were using statins in non-CKD group compared to 67.1% in CKD group (p < 0.01) (Table We compared the characteristics of patients with and without CKD. The average age was 64.3 ± 12.5 years in non-CKD group and 68.09 ± 12.94 years in CKD group (p = 0.05), elevated TG  and elevated FBG .The association between metabolic syndrome components and prevalence of CKD were explored Table . UnadjusAs diabetes plays a key role in the development of CKD, we also carried out sub-group analysis investigating whether metabolic syndrome components would still be associated with CKD in patients either with or without diabetes.p = 0.03). However, such association was not found in non-diabetes patients .The association between hypertension grades and CKD remained strongly significant in both unadjusted and multivariate adjusted models in diabetes patients , but also in patients without diabetes .Elevated TG was always independently associated with CKD, not only in patients with diabetes  or without diabetes .Reduced HDL-C was not associated with CKD in patients either with diabetes   or without diabetes , grade 1  to grade 2 . While only in hypertension grade 3, elevated TG was not associated with CKD   in the US, participants with metabolic syndrome had increased odds for chronic kidney disease compared to those without metabolic syndrome [ated FBG . In a reated FBG .Except that hypertension and diabetes have long been documented to be the major risks for the progression of CKD, elevated TG was found in our study to be associated with CKD, particularly independent of diabetes and hypertension grades. The mechanism of how the lipid level affect the injury of renal function was still incompletely understood. There are several mechanic pathways involved, including atherosclerosis, oxidative stress, chronic inflammation, hemodynamic change and endothelial dysfunction. For patients with dyslipidemia, specifically high TG, direct stimulation of glomerulosclerosis may be the key mechanism . In factTG has been strongly debated to be a causal factor driving atherosclerosis . There aThe main weakness of our study was the relatively small sample size with convenience sampling, by which the possibility of bias might exist. In addition, since it was a cross-sectional study rather than a causal design, it was difficult to tell the causal relationship between metabolic syndrome components and CKD. We could only conclude that there was a strong independent association between them.However, the strength of the study was that it was a multicenter cross-sectional study collecting data from all over the country, which made it possible for the conclusion to be generalized to all the patients with hypertension in China. Second, to our knowledge, this was the first study to evaluate the risk of CKD in patients with hypertension and other metabolic syndrome components. The importance of metabolic syndrome management in patients with hypertension should be further emphasized. Last, more attention should be focused on TG during risk management in order to prevent CKD and further research into the mechanism behind is needed.In our cross-sectional study of patients with hypertension, elevated TG, elevated FBG and hypertension grades were independently associated with the prevalence of CKD. Particularly, elevated TG was strongly associated with CKD, independent of diabetes and hypertension grades. Thus, among all the metabolic syndrome components, TG should also be concerned during the risk management of hypertension.Additional file 1. The ethics approval.Additional file 2. The patients' consent.


The mean (range) AUC0-t and AUC0-inf were 658.80 (268.29–1045.01) ng.h/ml and 676.98 (274.10–1050.96) ng.h/ml, respectively. The mean ke and t1/2 were 0.5386 h−1 and 1.91 h, respectively. The silymarin SMEDDS formulation soft capsule showed rapid absorption and high oral bioavailability.The present study aimed to determine the pharmacokinetic parameters and bioavailability of silymarin 140 mg SMEDDS formulation. An open-label, single-dose pharmacokinetic study was conducted. Twelve healthy volunteers were included in the study. After the volunteers had fasted overnight for 10 h, a single-dose generic silymarin 140 mg SMEDDS soft capsule was administered. Then 10 ml blood samples were taken at 0.0, 0.25, 0.50, 0.75, 1.0, 1.33, 1.67, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0, 10.0, and 12.0 h. The plasma silybin concentrations were analyzed using validated LC-MS/MS. The pharmacokinetic parameters were analyzed and calculated. The pharmacokinetic parameters were calculated after silymarin had been administered as a single capsule. The mean (range) C Silybum marianum or Carduus marianus) has been used for centuries as a treatment for liver-related disease [Silymarin, a flavonoid found in the seeds and fruit of milk thistle extraction  , 13, 14.2, were recruited for an open-label, single-dose under fasting conditions pharmacokinetics study. The inclusion criteria were that the volunteers had to be healthy according to their medical history, they had to undergo a physical examination, they had to be hepatitis B negative according to their clinical laboratory test results, they had to be nonsmoking, females had to be nonpregnant who were either unable to have children or committed to using acceptable nonhormonal contraceptives if potentially childbearing, and finally they had to be willing to comply with the study's procedures and restriction. Volunteers not included were those with a history of allergy to silymarin, a related structure of silymarin, or other components in the formulation; those who had a current or history of alcohol addiction or drug abuse; those who had been using any medication including vitamins, herbal products, and dietary supplements up to 14 days before or during the study; those who had consumed oranges, pomelos, or grapefruits up to 7 days before or during the study; and finally those who had ingested caffeine-containing beverages or food up to 3 days before or during the study. Any volunteers who had donated more than 300 ml of blood, suffered significant blood loss, or participated in other clinical trials up to 90 days before the start of the study were also excluded. All the volunteers were briefed on the details of the study. The volunteers who met the above criteria were eligible for participation after voluntarily signing their informed consent.Twelve healthy male and female volunteers, aged between 18 and 45 years old with a body mass index between 18.5 and 25.0 kg/mThe study was carried out at the Clinical Trial Unit (CTU) at the Faculty of Medicine at Chiang Mai University in Chiang Mai, Thailand, and complied with the International Conference on Harmonisation's Good Clinical Practice Guidelines. The Research Ethics Committee at the Faculty of Medicine at Chiang Mai University, Chiang Mai, Thailand, approved the study's protocol on 21st January 2016. Carduus marianus extract, equivalent to 140 mg silymarin, and calculated as 60 mg silybin. The SMEDDS was prepared using a mixture of Carduus marianus extract, polysorbate 80, polyoxyl-40-hydrogenated castor oil, polyethylene glycol 400, and propylene glycol. The recommendation dose of silymarin is a range from 70 to 140 milligrams three time per day in various disease conditions; thus one capsule of 140 mg silymarin was used for pharmacokinetic study.The silymarin SMEDDS formulation used in this study comprised Silyvercell® Soft Capsules manufactured by Phil Inter Pharma Co. Ltd. in Vietnam, which were made of 175 mgEvery volunteer who had fasted for at least 10 h and stayed at the CTU the night before the administration received a single oral dose silymarin SMEDDS capsule with 240 ml drinking water. Drinking water up to 1 h before or after taking the medication was not allowed. A standardized meal for lunch and dinner was served 4 and 10 h after the dose.2EDTA 0.0, 0.25, 0.50, 0.75, 1.0, 1.33, 1.67, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0, 10.0, and 12.0 h after the silymarin administration. They were then transported at a controlled temperature to the Pharmacy Service Center at the Pharmacy Faculty at Chiang Mai University, where the plasma samples were immediately separated and stored at −30 ± 5°C until their analysis.Ten ml blood samples were collected from each volunteer using a blood collection tube coated with KAn analytical method for measuring silybin in human plasma by LC-MS/MS was developed. The analytical procedures are briefly described as follows.μg/ml working solution of NRG  was subsequently added to 750 μl of plasma prepared for calibration standards, QC samples, and study samples. After adding 6 ml ethyl acetate, the mixture was mixed using Vibramax 110 shaker at 1500 rpm for 15 min at room temperature and then centrifuged using a Hermle Z 383K at 3000 rpm for 1 min at 20°C. A 4 ml portion of organic phase was collected, transferred into another test tube, and evaporated to dryness under a stream of 5 psi nitrogen gas for 40 min at 25°C. The dried residue was dissolved with 200 μl of mobile phase and shaken using a Vibramax 110 shaker at 2000 rpm for 5 min at room temperature. An aliquot was centrifuged at 11000 rpm for 5 min at 25°C. The supernatants were collected in an insert vial and 10 μl was injected into the analytical column.Sample Preparation. Thirty microliters of 7 Analytical Method. The LC-MS/MS analysis was carried out using an HPLC Agilent 1260 system consisting of a model 1260 Bin solvent delivery unit, a 1260 Hip on-line degasser, a 1260 Hip ALS autoinjector, and a 1260 TCC column oven  in combination with an API 3200 mass spectrometer .μm); it was fitted with a Mightysil RP-18 GP guard column  purchased from Fortis Technologies Ltd. (United Kingdom) and the Thermo Electron Corporation (United States), respectively. The column thermostat was at a controlled temperature of 25°C. The flow rate was set at 0.5 ml/min. The elution was carried out using a mixture of methanol and 0.125% ammonia solution (18:82 v/v), filtered through a 0.2 μm cellulose acetate membrane filter  before use. The injection volume was 10 μl.LC Conditions. The chromatographic analysis of the sample was performed on a Fortis C18 column  mode and was equipped with an electrospray ionization (ESI) source. Quantification was achieved with MS/MS detection in negative ion mode. The analyte (SLB) and internal standard (NRG) were monitored at transition m/z 481.007→125.110 and 270.932→150.912, respectively. The ion spray voltage was set at −4500 V. The nebulizer gas (GS1), auxiliary gas (GS2), curtain gas (CUR), and collision gas (CAD) were set at 60, 55, 35, and 6 psi, respectively. The declustering potential (DP), entrance potential (EP), collision cell entrance, potential (CEP), collision energy (CE), and collision cell exit potential (CXP) were −50, −8.5, −18, −42, and −6 V for the SLB and −40, −10, −16, −11, and −6 V for the NRG.The analytical method for determination of silybin in human plasma by LC-MS/MS was developed and validated according to EMA and US FDA guidance including precision, accuracy, selectivity, linearity, recovery, and stability.max, tmax, AUC0-t, AUC0-inf, ke, t1/2, CL, and Vd. Cmax and tmax came from the actual data. The trapezoidal rule (linear up log down) was used to calculate AUC0-t. AUC0-inf was estimated from AUC0-t, the last plasma concentration (Clast), and the elimination rate constant (ke). The elimination rate constant (ke) was calculated from −2.303 multiplied by the slope of the linear regression equation curve of the log-transformed plasma drug concentrations at the terminal log linear phase. The elimination half-life (t1/2) was calculated as 0.693 divided by ke. The equations for calculating Vd and CL were dose/(λz.AUCinf_obs) and dose/AUCinf_obs, respectively. The pharmacokinetic parameter determination was performed using Phoenix™ Winnonlin® 6.3 computer software.The pharmacokinetic parameters were analyzed including C0-t, AUC 0-inf, and Cmax was reported. The data from all the volunteers participating in the entire study were used for the pharmacokinetic parameters and statistical analysis.The pharmacokinetic data were expressed as mean ± SD. A ninety-five percent confidence interval (95% CI) for the mean of AUC 2 (a range of 19.8–24.1 kg/m2).Twelve healthy Thai volunteers consisting of 6 males and 6 females took part in the study. The mean age ± SD of the volunteers was 25.8 ± 5.2 years (a range of 20–35 years). The mean weight ± SD of the volunteers was 61.4 ± 5.5 kg (a range of 51.0–71.7 kg). The mean height ± SD of the volunteers was 165.4 ± 7.6 cm (a range of 156–180 cm). The mean BMI ± SD of the volunteers was 22.4 ± 1.3 kg/mThe average plasma silybin concentration of the silymarin SMEDDS soft capsules is shown in max, tmax, AUC0-t, AUC0-inf, ke, t1/2, CL, and Vd. The pharmacokinetic parameters, i.e., tmax, Cmax, AUC0-t, and AUC0-inf, are collated in max value was 812.43 (259.47–1505.47) ng/ml and the mean (range) AUC0-t value was 658.80 (268.29–1045.01) ng.h/ml. The mean (range) AUC0-inf value was 676.98 (274.10–1050.96) ng.h/ml. The mean (range) tmax value was 0.80 (0.25–1.67) h.The pharmacokinetic parameters were calculated by a noncompartmental model using Phoenix Winnonlin 6.3 software. These parameters include Ce) was calculated based on the log-transformed plasma drug concentration-time curve at the terminal phase of elimination. The time points used for the linear regression analysis in the ke calculation were selected automatically by the Phoenix Winnonlin 6.3 computer software, according to the “Best Fit” method. The regression analysis parameters ke and t1/2 are presented in e value was 0.5386 h−1. The average t1/2 value was 1.91 h.The elimination phase constant  for the lower and upper mean for Cmax (0.54 ± 0.19 versus 1.06 ± 0.48 h) (p<0.05).Pharmacokinetic parameters show no difference between male and female volunteers except for tAdverse events (AEs) were followed and recorded according to the interview data and physical examination results. With regard to the safety of subjects, adverse events were observed and concomitant drug assessments were performed throughout the study. The volunteers' blood pressure, pulse rate, respiratory rate, and body temperature were within the normal range determined at their screening and during the entire study period. Their tolerance to formulations was good. There were no adverse events/serious adverse events throughout this study.A simple, sensitive, and specific LC-MS/MS method was developed and validated for determination of silybin in human plasma. Sufficient separation of silybin peak and naringenin  peak from other interferences was observed. The calibration curve for the analyte was linear in the range of 4–2000 ng/ml with coefficient of determination ≥ 0.99. The lower limit of quantification for silybin was 4 ng/ml. The within-run and between-run accuracy and precision were within the acceptable ranges. At room temperature, silybin was stable in plasma for at least 5 hours, and for at least 37 days at −30 ± 5 degree Celsius. The method has been found to be highly precise, accurate, robust, and suitable for the determination of silybin in plasma samples.max after the oral administration of silybin in a SMEDDS capsule was approximately 2 times faster than in a conventional capsule. Moreover, the Cmax and AUC were approximately 4.9–5.9 and 1.7–2.5 times higher than a conventional capsule. In addition, if compared to a different commercial preparation of a powdered milk thistle extract capsule [max and AUC of silybin from the silymarin SMEDDS soft capsules were approximately 34–62 and 9.5–26.3 times higher, respectively, than from the powdered milk thistle extract capsule. However, there were no previous reports about pharmacokinetic, bioavailability, and bioequivalence study of silymarin in the form of SMEDDS; thus pharmacokinetic data cannot be compared to the present study.The silymarin SMEDDS formulation in previous animal study has shown the enhancing bioavailability properties after oral administration , 14, bei capsule , 18, theThe elimination half-life of silybin in humans was previously reported as being 1.3–7.8 h after the administration of various forms of silymarin formulation and was shown to be within the same range in this study (0.73–7.50 h).The above data indicated that after oral administration of silybin in a SMEDDS formulation, the absorption of the silybin was faster, and the extent of the absorption was enhanced. However, further study with the same group of volunteers using a noninferior test is recommended for a more appropriate comparison. It would be more beneficial to conduct study on hepatic disease patients who might have difference in pharmacokinetic parameters.max value was 812.43 (259.47–1505.47) ng/ml and the average AUC0-t value was 658.80 (268.29–1045.01) ng.h/ml. The average AUC0-inf value was 676.98 (274.10–1050.96) ng.h/ml. The mean (range) tmax value was 0.80 (0.25–1.67) h. It showed a fast absorption (tmax < 1 h) and high oral bioavailability . The subjects tolerated the study well, as no clinically significant or serious adverse drug reactions were noticed.After a single oral dose of silymarin SMEDDS soft capsules, the mean (range) C


Antioxidant activity was performed using 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals and ferric reducing/antioxidant power (FRAP) methods. The α-Glucosidase and β-Galactosidase inhibitory activities were investigated using an in vitro model. Moreover, phytochemical analysis of tested extracts was carried out. The aqueous fraction of this herb exhibited the highest antioxidant activity for both DPPH and ABTS methods, IC50=125.40±2.40 μg/mL and IC50=65.23±2.49 μg/mL, respectively. However, the ethyl acetate fraction possessed the strongest inhibitory effect towards α-Glucosidase (IC50=1.112±0.026 mg/mL). Furthermore, the result showed high levels of phenolic content. The results showed that this plant could be a significant source of medically important natural compounds.Oxidative stress plays a major role in diabetic physiopathology; hence, the interest of using natural antioxidants as therapeutic tools exists. The aim of this study was the evaluation of Medicinal plants are one of the main resources of therapeutic agents. Indeed, 80% of the world's population uses plants in health care . Recentl Aristolochia longa (A. longa) is a medicinal plant belonging to Aristolochiaceae family, which is widely distributed in the tropical and temperate regions [ A. longa locally known as “Barraztam” is a species commonly used in Moroccan traditional medicine. Many traditional healers also use a small amount of its rhizome powder with honey or salted butter for the treatment of abdominal pain and upper respiratory tract infections [Among Moroccan medicinal plants, regions . Aristol regions . A. longfections –10.α-Glucosidase and β-Galactosidase [ in vitro antioxidant activity, α-Glucosidase and β-Galactosidase inhibitory potentials of A. longa root extracts.Likewise, diabetes mellitus is a major cause of mortality and the most common metabolic disorder characterized by hyperglycemia due to lack of insulin production by the pancreas or the inability of the insulin produced to control blood glucose , 12. Onetosidase , 14. In p-Nitrophenyl-α-D-glucopyranoside, 2-Nitrophenyl-β-D-galactopyranoside, α-Glucosidase from Saccharomyces cerevisiae, β-Galactosidase from Aspergillus oryzae, Acarbose, Folin-Ciocalteu reagent, rutin, catechin, 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and ascorbic acid were purchased from Sigma-Aldrich (France). All other reagents were of analytical grade. A. longa were collected in April 2016, in the province of Al Haouz in Morocco. The collected plant materials were authenticated at the Herbarium of Botany Department of the Scientific Institute of Rabat, Morocco. The roots of the plant were washed, dried at room temperature from 48 to 92 h, grounded into powder, and then stored in glass bottles preserved from light and moisture until use.Roots of A. longa dry roots powder. After 24 h of maceration under magnetic stirring at room temperature, the mixture was centrifuged. Then, all extracts already prepared were filtered through a filter paper (Whatman) and evaporated to dryness by a rotary evaporator at 50°C. The extracts obtained were kept at 4°C until further uses.To prepare the extracts, the technique of continuous hot extraction by a Soxhlet extractor was carried out using solvents of different polarities. Briefly, 10 g of root powder was extracted successively with 100 mL each of ethyl acetate, methanol, and water until the extracts were colorless in the siphon tube. The aqueous extract was prepared by adding 500 mL of distilled water to 50 g of A. longa extracts was assessed by Folin-Ciocalteu method [Total phenolic content ofu method  as descru method . Gallic  A. longa extracts was determined according to the method described by Dewanto et al. [The total flavonoid content ofo et al.  using Al A. longa extracts was determined as reported by Julkunen-Tiitto [The proanthocyanidin content ofn-Tiitto . Catechi50 was calculated for comparing the obtained results.Radical scavenging activity of the extracts was measured using the stable radical DPPH such as that determined by Sayah et al.  with som∙+ was carried out according to the method described by Sayah et al. [∙+) was prepared by the reaction between 10 mL of ABTS (2 mM) in H2O and 100 μL of potassium persulfate (K2S2O8) (70 mM). The mixture was incubated in the dark for 16 hours at room temperature. Then, the ABTS∙+ solution was diluted with methanol to obtain an absorbance of 0.70 at 734 nm. Then, 200 μL of each extract was mixed with 2 mL of the diluted ABTS∙+ solution and allowed to react for 1 minute. After, the absorbance of the ABTS∙+ radical is measured at 734 nm. All samples were made in triplicate. The results were represented as Trolox equivalent per gram of extract dry weight (mg TE/edw).The TEAC test  or discoloration test of ABTSh et al. . Briefly3 . The absorbance was measured at 700 nm. The results are expressed as ascorbic acid equivalent per gram of extract dry weight (mg AAE/g edw).The FRAP test is performed according to the method described by Sayah et al. . Brieflyα-Glucosidase inhibitory activity was monitored using the substrate p-Nitrophenyl α-d-glucopyranoside (pNPG), which is hydrolyzed by α-Glucosidase to release p-Nitrophenyl (a colored agent which can be monitored at 405 nm), according to the method described by Marmouzi et al. [50) were determined.The i et al. , 21. Theβ-Galactosidase inhibitory activity was assessed according to the method of Bouabid et al. [β-D-Galactopyranoside as substrate, which is hydrolyzed by β-Galactosidase to release 2-nitrophenyl . Briefly, a mixture of 150 μL of the samples at different concentrations (0.5-5 mg/mL) and 100 μL of sodium phosphate buffer 0.1 M (pH=7.6) containing the enzyme β-Galactosidase solution (0.1U/mL) was incubated at 37°C for 10 min. After preincubation, 200 μL of gala solution 1 mM in sodium phosphate buffer 0.1 M (pH=7.6) was added. The reaction mixtures were incubated at 37°C for 30 min. After incubation, 1 mL of 0.1 M of Na2CO3 were added to stop the reaction and the absorbance was recorded at 405 nm using the spectrophotometer. The β-Galactosidase inhibitory activity was expressed as percentage inhibition and calculated using the same formula as the α-Glucosidase test, and the IC50 values were determined. Quercetin was used as positive control and the experiment was carried out in triplicate.The d et al.  using 2-p < 0.05 was considered to be statistically significant. All analyses were performed with GraphPad Prism 6. Also, IC50 values were determined using a nonlinear regression curve with the same program.Results are expressed as a mean ± standard error of the mean. Differences between the means were determined by one-way analysis of variance (one-way ANOVA). A difference in the mean values of  A. longa were found to be 32.55±0.43 mg GAE/g edw, which are significantly (p< 0.05) greater than methanolic fraction and aqueous extract (24.48±1.63 mg GAE/g edw and 13.13±0.48 mg GAE/g edw) respectively. There is no significant difference between EAF and aqueous fraction (AF) (28.90±1.36 mg GAE/g edw). Results of flavonoid and proanthocyanidin contents show that ethyl acetate fraction resulted also in significantly (p< 0.05) higher values of those compounds  followed by the aqueous fractions that were 7.60±0.11 mg RE/g edw and 10.79±0.49 mg CE/g edw, respectively. The interesting phenolic contents of this herb indicate an important health promoting activity. Indeed, Djeridane et al. [ A. longa roots and they estimated the total phenolics and flavonoids content as 1.47±0.02 mg GAE/g edw and 0.81±0.02 mg RE/g edw, respectively [ A. bracteolata and A. indica, revealed that the methanol extract from each plant contains high amount of phenols and flavonoids [Total phenolic, flavonoid, and proanthocyanidins contents are presented in e et al.  worked oectively . These cectively . Previouavonoids , 26. Pheavonoids . in vitro and in vivo antioxidant activity have been developed, but only a few fast and reliable methods for a large number of plant extract samples exist [Recently, many scientific studies have shown that free radicals play a major role in the development of cancer, heart disease, aging, cataracts, and immune system damage . These ues exist –34. Furt in vitro antioxidant activity of plant extracts [ A. longa at various concentrations. All A. longa extracts showed scavenging effect, which increases with the concentration of samples. At 416 μg/mL concentration, aqueous and methanolic fraction of A. longa exhibited increased DPPH radical scavenging activity of 77.17% and 74.66%, respectively. Moreover, aqueous extract showed less activity at all concentrations. These results showed that A. longa roots contained high amount of radical scavenging compounds with proton-donating ability. Similar result was observed in one previous study of Benmehdi et al. [ A. clematitis roots.DPPH is one of the stable free radicals, commercially available, nitrogen centered, and largely used for evaluating scavenging activity of antioxidant standards and plant extracts with a characteristic absorbance at 517 nm , which dextracts , 41. Recextracts –44. Fromi et al. , which sμg/mL concentration, aqueous and methanolic fraction of A. longa possessed 90.89% and 82.58% scavenging activity on ABTS. At the concentrations investigated, this effect may indicate the capacity of the herb to minimize oxidative damage to certain vital tissues of the body [The radical cation of 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) is stable in its free form. The concentration of this radical can be determined by measuring the absorbance at 734 nm. The addition of an antioxidant to a solution of this radical leads to its reduction and a decrease in absorbance. This decrease depends on the antioxidant activity of the test compound, but also on the time and the concentration . The expthe body . These rthe body  using thμg/mL concentration and among the solvents tested, aqueous fraction exhibited higher reducing activity 80.73%. The results obtained are consistent with the studies carried out on A. longa  [ A. indica  [In the FRAP method, antioxidants present in the sample reduced the Fe3+/ferricyanide complex to the blue ferrous form , which cal part)  and A. ial part)  and  , which i50 are calculated as shown in 50 indicates greater antioxidant activity. Indeed, the aqueous fraction showed the highest DPPH radical inhibition value (IC50=125.40±2.40 μg/mL), while the methanolic fraction was the most active against the ABTS radical cation (IC50=61.58±2.15 μg/mL), and the Trolox value for both tests was 1.79±0.35 μg/mL and 0.70±0.01 μg/mL, respectively. Our results are in analogy to those of Ramalalingam et al. [ A. indica, who showed IC50 value 182.31±0.31 μg/mL, while standard showed the value of 30.12±0.11 μg/mL [ A. longa roots had the weakest activity compared to Trolox, among all the plants tested [ A. bracteolate who found that the methanolic extract had potent antioxidant activity [p <0.05); this result can be interpreted by the fact that the extracts have several compounds, while the concentrations of those who could perform this activity will be really low. It is also noted that the antioxidant effects change depending on the test used. In fact, the antioxidant activity depends on the interactions in the reaction media between the substrate(s)  and the active molecule(s) that trap them. Other studies have shown that the roots of A. longa possess antioxidant activity [50 values of fruit aqueous extract and fruit methanol extract were found to be 145.15±0.78 μg/mL and 186.21±6.24 μg/mL, respectively. The difference between the results is due, as already mentioned, to the difference of the phytoconstituents, also to the method used, the harvest period, and the studied part To compare the results, the ICm et al.  using th11 μg/mL . In anots tested . Furtheractivity . The effactivity –58. Phenactivity , 60. Plaactivity . Moreoveactivity , and we  A. longa are evaluated for their antidiabetic activity. Two different in vitro assays were used to evaluate this activity, α-Glucosidase and β-Galactosidase uptake assay.Diabetes is characterized by high blood sugar levels, which can lead to serious complications, so the goal of treating patients with diabetes is to maintain near-normal levels of glycemia control. In modern medicine, there is no treatment or medication to treat diabetes without side effects, which are related to the use of insulin and oral hypoglycemic agents . Medicinα-Glucosidase catalyzes the final step in the digestion of carbohydrates and is located in the brush-border surface membrane of intestinal cells [al cells . Its inhal cells . This isal cells .β-Galactosidase catalyzes the hydrolysis of β-Galactosides subtracts to simple carbohydrates in the intestine. Subsequently, the inhibition of this enzyme can lead to the intestinal hydrocarbon reduction and eventually decreases the glucose level. A. longa extracts showed inhibitory effects on both enzymes tested as presented in α-Glucosidase and β-Galactosidase activity concentration dependently (0.5–5 mg/mL). Indeed, at the concentration of 1.5 mg/mL, the ethyl acetate fraction has the highest inhibitory activity against α-Glucosidase (76.56±2.54%) and β-Galactosidase (12.70±1.27%). The methanolic fraction showed a moderate inhibition of α-Glucosidase (21.94±1.34%) and β-Galactosidase (2.05±1.22%). However, the aqueous extract inhibited only the enzymatic activity of β-Galactosidase (2.20%). These results are consistent with those of Janani and Revathi, [A. indica). Their study revealed that the methanolic extract of whole plant showed α-Glucosidase inhibitory activity, which increased with the increasing concentration.On the other hand, Revathi,  who work50 which represents the concentration of an inhibitor that is required for 50% inhibition of its targeted enzyme. The fractions of ethyl acetate and methanol showed a strong inhibitory capacity against α-Glucosidase with IC50 values of 1.112±0.026 and 2.378±0.037 mg/mL, respectively. These inhibition values are greater than that obtained by Acarbose (0.199±0.014 mg/mL), used as standard antidiabetic. Similar effects were observed on A. indica [α-Glucosidase inhibition values by the aqueous extract and aqueous fraction are above 5 mg/mL. For β-Galactosidase inhibitory capacity, all extracts showed an IC50 value greater than 5 mg/mL. The fact that α-Glucosidase and β-Galactosidase showed difference is due to structural differences related to the origins of enzymes [To measure the inhibitory effectiveness of each extract, we used the IC. indica . Likewis enzymes . A. longa extracts against the enzyme α-Glucosidase demonstrate their potential abilities to reduce the postprandial increase of blood glucose levels in diabetic patients and their capacities to prevent type 2 diabetes. Hence, it is suggested that the mechanism of antihyperglycemic may be due to the antioxidant activity of this herb. Our finding is in accordance with earlier reports that showed that, in animal models, two of its species, aerial parts of A. indica [ A. ringens roots extracts [α-Glucosidase inhibition including a species of Aristolochiaceae (Asarum heterotropoides) [ A. longa demonstrated its abilities to produce a high amount of phenolic compounds and several flavonoids, including alkaloids [α-Glucosidase in both in vitro and in vivo studies [α-Glucosidase [ A. longa may be associated with the presence of other phytoconstituents like alkaloids, tannins, and saponins [α-Glucosidase, have fewer side effects and are less expensive compared to synthetic pharmacotherapeutics like Acarbose [ Aristolochia sp from different parts of the world are not only due to the type of specie but also different agroclimatic conditions, extraction method, harvest period, and characterization techniques [ A. longa, which is used in traditional medicine for the treatment of several diseases. This herb was not previously investigated for its in vitro antidiabetic activity. The results from this work should be relevant to the human body.The inhibitory effects of. indica  and the extracts , showed opoides) . The diflkaloids , saponinlkaloids . The phelkaloids . Moreove studies  and may  studies . Importacosidase . The inhsaponins , 77. Thesaponins . These cAcarbose , and theAcarbose . Generalchniques , 82 and chniques . In the  A. longa roots had the best antioxidant effects against the DPPH and ABTS radicals and a strong ferric reducing power. This suggests that A. longa can be used to prevent and control the oxidative stress induced by free radicals. The antidiabetic activity was also investigated, focusing on the inhibitory effects on α-Glucosidase and β-Galactosidase. Our study is the first to report a potential mode of action of A. longa and suggests that the effect of this plant is due to the inhibition of digestive enzymes. On the other hand, the presence of flavonoids and phenols concludes that this herb has multiple biological properties. Other studies must be conducted to isolate the active ingredients of this plant, identify them, and study their bioactivity.The aqueous fraction of the


Plagiorhegma dubium (Berberidaceae) has been listed as an endangered species in Korea due to extensive collection and destruction of natural habitats. In this study, 30 microsatellite loci, including 25 polymorphic loci, were developed for P. dubium for use in population‐level genetic analyses.P. dubium using Illumina HiSeq high‐throughput sequencing. To test utility of the developed markers, we genotyped 60 individuals from three populations and estimated the number of alleles and levels of observed and expected heterozygosity. Expected heterozygosity levels ranged from 0.000 to 0.594, 0.000 to 1.000, and 0.000 to 0.744 in the three populations, respectively.We carried out transcriptome sequencing and isolated a total of 30 expressed sequence tag–simple sequence repeat markers from P. dubium. Population genetic studies with these markers will provide valuable insights for conservation by unraveling evolutionary patterns of P. dubium.These transcriptome‐derived simple sequence repeat markers are highly polymorphic and can be widely used in characterization of the endangered  Plagiorhegma Maxim. (Berberidaceae) is composed of only one species, P. dubium Maxim., which has a narrow distribution in northeastern Asia and Siberia , 2 μL of 2.5 mM dNTPs, 0.1 μM of forward and reverse primer, 0.1 μL of TaKaRa Ex Taq, and 5–10 ng of template DNA. All PCRs were performed in a GeneAmp PCR System 9700 thermocycler  using the following protocol: initial denaturation at 98°C for 5 min; followed by 30 cycles of denaturation at 95°C for 1 min, annealing at locus‐specific annealing temperature , Incheon, Republic of Korea (Appendix A), observed heterozygosity, and expected heterozygosity using GenAlEx 6.5 . A total of 30 markers were successfully amplified, of which polymorphism was detected in 25  protein database with BLASTX E‐value  × 10−5. P. dubium  Sequence Read Archive (Bioproject ID PRJNA472226). Sequence information for the developed primers has been deposited to NCBI; GenBank accession numbers are provided in Table