Point-of-care testing of methamphetamine and cocaine utilizing wearable sensors.

The imperative for the point-of-care testing of methamphetamine and cocaine in drug abuse prevention necessitates innovative solutions. To address this need, we have introduced a multi-channel wearable sensor harnessing CRISPR/Cas12a system. A CRISPR/Cas12a based system, integrated with aptamers specific to methamphetamine and cocaine, has been engineered. These aptamers function as signal-mediated intermediaries, converting methamphetamine and cocaine into nucleic acid signals, subsequently generating single-stranded DNA to activate the Cas12 protein. Additionally, we have integrated a microfluidic system and magnetic separation technology into the CRISPR system, enabling rapid and precise detection of cocaine and methamphetamine. The proposed sensing platform demonstrated exceptional sensitivity, achieving a detection limit as low as 0.1 ng/mL. This sensor is expected to be used for on-site drug detection in the future.

External application of brassinolide enhances cold resistance of tea plants (Camellia sinensis L.) by integrating calcium signals.

MAIN CONCLUSION: Exogenous brassinolide can activate the expression of key genes in the calcium signalling pathway to enhance cold resistance of tea plants. Brassinolide is an endogenous sterol phytohormone containing multiple hydroxyl groups that has the important function of improving plant cold resistance and alleviating freeze damage. To explore the molecular mechanism of how brassinolide improves the cold resistance of tea plants, "Qiancha 1" was used as the material, and the method of spraying brassinolide on the leaves was adopted to explore its effects on the tea plants under 4 °C low-temperature treatment. The results showed that brassinolide can significantly increase the protective enzyme activity of tea plants under cold stress and reduce cold damage. At the transcriptome level, brassinolide significantly enhanced the expression of key genes involved in calcium signal transduction, Calmodulin (CaM), Calcium-dependent protein kinase (CDPK), calcineurin B-like protein (CBL) and calmodulin-binding transcriptional activators (CAMTA), which then activated the downstream key genes transcriptional regulator CBF1 (CBF1) and transcription factor ICE1 (ICE1) during cold induction. Quantitative real-time PCR (qRT‒PCR) results showed that the expression of these genes was significantly induced after treatment with brassinolide, especially CaM and CBF1. When calcium signalling was inhibited, the upregulated expression of CBF1 and ICE1 disappeared, and when CAMTA was knocked down, the expression of other genes under cold stress was also significantly reduced. The above results indicate that brassinolide combined with the calcium signalling pathway can improve the cold resistance of tea plants. This study provides a new theoretical basis for the study of the cold resistance mechanism of brassinolide.

Research method of rapid determination of chiral pesticide fenpropathrin enantiomers in fruit and vegetable puree by supercritical fluid chromatography.

A method is first established for the separation and determination of fenpropathrin enantiomer residues in apple puree, strawberry puree, and tomato puree considered a supplementary food for infants by supercritical fluid chromatography. After the sample was extracted with acetonitrile and cleaned up by a solid-phase extraction column, then it was separated by a CHIRALPAK AD-3 chiral column with gradient elution at a flow rate of 1.5 mL/min using methanol and supercritical carbon dioxide as the mobile phase, detected by ultraviolet detector at 230 nm wavelength and quantified with the external standard method. The limits of quantification of the two fenpropathrin enantiomers were both 0.2 mg/kg, the linear ranges were 1.0-20.0 mg/L with linear correlation coefficients greater than 0.9992, the recoveries in the spiked samples at 0.2, 0.4 and 2.0 mg/kg were from 80.6 to 105%, and the relative standard deviation reached 2.6-7.7%. This method has the advantages of convenient operation, good resolution, and environmental protection, which can satisfy the requirement of determination for fenpropathrin enantiomer residues in fruit and vegetable puree as supplementary food for infants.

Hydroxytyrosol and Oleuropein Inhibit Migration and Invasion via Induction of Autophagy in ER-Positive Breast Cancer Cell Lines (MCF7 and T47D).

Hydroxytyrosol (HT) and oleuropein (OL), the most abundant of the phenolic compounds in olives, have anticancer properties against breast cancer (BC). However, little attention has been paid to the mechanism of HT or OL in BC cells. The objective of this study was to identify the underlying molecular mechanisms of these compounds. ER-positive BC MCF7 and T47D cells were treated with HT and OL in combination with hepatocyte growth factor (HGF), rapamycin (Rapa, an agonist of autophagy) or 3-methyladenine (3-MA, an inhibitor of autophagy). Cell viability, metastasis capability and autophagy-related proteins were evaluated by wound healing assays, Transwell assays and Western blot. HT and OL reduced the cell viability of MCF-7 and T47D cells in a dose-dependent manner. Both cells were more sensitive to HT than OL. In addition, Rapa significantly inhibited HGF-induced migration and invasion, indicating that metastases of both BC cells could be inhibited by suppression of autophagy. Moreover, HT and OL significantly blocked HGF- or 3-MA-induced cell migration and invasion by reversing LC3II/LC3I and Beclin-1 downregulation and p62 upregulation. These findings revealed that HT and OL could suppress migration and invasion by activating autophagy in ER-positive BC cells, which might be a promising therapeutic strategy.

A nomogram prediction model for postoperative seroma/hematoma in elderly subjects after TAPP.

BACKGROUND: Formation of seroma/hematoma is one of the most common postoperative complications following laparoscopic inguinal hernia repair. This study aimed to identify risk factors associated with seroma/hematoma and construct a prediction model. METHODS: Elderly subjects undergoing laparoscopic Transabdominal preperitoneal Patch Plasty (TAPP) were included in this study. The observation endpoint was set as the occurrence of seroma/hematoma within 3 months after TAPP surgery. Independent risk factors were identified through preliminary univariate screening and binary logistic regression analysis. These risk factors were then used to construct a nomogram predictive model using R software. RESULTS: A total of 330 patients were included in the analysis, of which 51 developed seroma/hematoma, resulting in an incidence rate of 15.5%. Obesity (OR: 3.54, 95%CI: 1.45-8.66, P = 0.006), antithrombotic drug use (OR: 2.73, 95%CI: 1.06-7.03, P = 0.037), C-reactive protein (CRP) ≥ 8 (OR: 2.72, 95%CI: 1.04-7.10, P = 0.041, albumin/fibrinogen ratio (AFR) < 7.85 (OR: 2.99, 95%CI: 1.28-7.00, P = 0.012), and lymphocyte/monocyte ratio (LMR) < 4.05 (OR: 12.62, 95%CI: 5.69-28.01, P < 0.001) were five independent risk factors for seroma/hematoma. The nomogram model has well predictive value for seroma/hematoma, with an AUC of 0.879. CONCLUSIONS: The nomogram model based on obesity, antithrombotic drug, CRP, AFR, and LMR has a proved good predictive value and it has potential in clinical practice.

Biodegradable bio-film based on Cordyceps militaris and metal-organic frameworks for fruit preservation.

In this study, Cordyceps militaris matrix was employed for the first time to fabricate a biodegradable food packaging. Carmine and Ag@CuBTC were introduced to cross-link with mycelium and were uniformly dispersed within the matrix to enhance the water resistance, antimicrobial, and antioxidant properties of the bio-films. The bio-film displayed high biodegradability, with nearly 100 % degradation achieved after three weeks. The bio-film exhibited exceptional resistance to oxidation (49.30 % DPPH and 93.94 % ABTS•+), as well as effective inhibitory capabilities against E. coli and S. aureus, respectively. The composite film maintained a high CO2/O2 selective permeability, which was advantageous for mitigating fruit metabolism and extending shelf life. Simultaneously, food preservation experiments confirmed that these bio-films can decelerate the spoilage of fruits and effectively prolong the shelf-life of food. The experimental findings indicated that the prepared Bio-R-Ag@Cu film held promise as an environmentally friendly biodegradable material for food packaging.

Modified Frailty Index and Albumin-Fibrinogen Ratio Predicts Postoperative Seroma After Laparoscopic TAPP.

Background: Postoperative seroma is the most common minor complication after inguinal hernia repair surgery and can have negative consequences. The objective of this study was to identify potential risk factors for postoperative seroma. Methods: This study consecutively included 354 elderly patients with inguinal hernia who underwent laparoscopic Transabdominal preperitoneal Patch Plasty (TAPP). Seroma diagnosis was conducted by the same experienced surgeon based on the physical examinations combined with ultrasound. Risk factors for seroma were identified through univariate analysis and subsequently included in the binary multivariate logistic regression model. Results: A total of 40 patients experienced postoperative complications of seroma, with an incidence rate of 11.3% (40/354). The binary logistic regression analysis revealed that obesity (OR: 2.98, 95% CI: 1.20-7.41, P = 0.018), disease duration ≥ 4.5 years (OR: 4.88, 95% CI: 2.14-11.18, P < 0.001), albumin-fibrinogen ratio (AFR) level < 9.25 (OR: 6.13, 95% CI: 2.00-18.76, P = 0.001), and modified frailty index (mFI) score ≥ 0.225 (OR: 6.38, 95% CI: 2.69-15.10, P < 0.001) were four independent risk factors for postoperative seroma. Conclusion: Obesity, prolonged disease duration, decreased AFR level, and increased mFI score independently predict postoperative seroma after laparoscopic TAPP.

[Determination of four fatty acid ethyl esters in olive oil by solid phase extraction-gas chromatography].

The fatty acid ethyl ester (FAEE) content of olive oil is an important indicator of its quality. At present, the international standard method used to detect FAEEs in olive oil is silica gel (Si) column chromatography-gas chromatography (GC); however, this technique presents a number of disadvantages, including complex operation, long analysis times, and high reagent consumption. In this study, a method based on Si solid phase extraction (SPE)-GC was established to determine four FAEEs in olive oil, namely, ethyl palmitate, ethyl linoleate, ethyl oleate, and ethyl stearate. First, the effects of the carrier gas were investigated, and He gas was ultimately selected as the carrier gas. Next, several internal standards were screened, and ethyl heptadecenoate (cis-10) was determined as the optimal internal standard. The SPE conditions were also optimized, and the effects of different brands of Si SPE columns on the recoveries of analytes were compared. Finally, a pretreatment method in which 0.05 g of olive oil was extracted with n-hexane and purified through a Si SPE column (1 g/6 mL) was developed. A sample could be processed within approximately 2 h using a total reagent volume of about 23 mL. Validation of the optimized method revealed that the four FAEEs have good linearities within the range of 0.1-5.0 mg/L (coefficients of determination (R2)>0.999). The limits of detection (LODs) of the method were within 0.78-1.11 mg/kg, and its limits of quantification (LOQs) were in the range of 2.35-3.33 mg/kg. The recoveries ranged from 93.8% to 104.0% at all spiked levels tested (4, 8, and 20 mg/kg), and the relative standard deviations were 2.2%-7.6%. Fifteen olive oil samples were tested using the established method, and the total FAEEs of three extra-virgin olive oil samples were found to exceed 35 mg/kg. Compared with the international standard method, the proposed method has the advantages of simpler pretreatment process, shorter operation time, lower reagent consumption and detection cost, high precision, and good accuracy. The findings provide an effective theoretical and practical reference for improving olive oil detection standards.

[Simultaneous determination of monochloropropanediol esters and glycidyl esters in vegetable oils by acidic transesterification-gas chromatography-mass spectrometry].

A comprehensive analytical method based on gas chromatography-mass spectrometry (GC-MS) was developed for the determination of 3-monochloropropanediol esters, 2-monochloropropanediol esters, and glycidyl esters in vegetable oils. Different parameters, such as bromination reaction temperature, bromination reaction time, derivatization reagent dosage, and derivative reaction time, were studied. The optimal conditions were as follows: 0.25 g of oil was weighed in a 10-mL glass tube, followed by the addition of 2 mL tetrahydrofuran, 25 µL of internal working standard solutions, and 30 µL of acid aqueous solution of NaBr, homogenized, and the mixture was incubated at 50 ℃ for 15 min. The reaction was stopped by the addition of 3 mL of an aqueous solution of sodium hydrogen carbonate. To separate the oil from the water phase, n-heptane was added, and the upper layer was transferred to an empty test tube and evaporated to dryness under a nitrogen stream. The residue was dissolved in 1 mL of tetrahydrofuran. 1.8 mL of sulfuric acid solution in methanol was added to the sample, and the resulting mixture was incubated at 40 ℃ for 16 h. The reaction was stopped by the addition of 0.5 mL of an aqueous solution of sodium hydrogen carbonate. After purification by n-hexane and derivatization of phenylboric acid, the derivatives were extracted with n-hexane. After nitrogen blowing, the residue was dissolved in 1 mL of n-hexane, and then filtered through a 0.45-µm membrane filter unit prior to GC-MS analysis. Temperature programming was applied at an initial temperature of 80 ℃. After 0.5 min, the temperature was raised to 180 ℃ at a rate of 20 ℃/min, held for 0.5 min, raised to 200 ℃ at a rate of 5 ℃/min for 4 min, and finally raised to 300 ℃ at a rate of 40 ℃/min for 4 min. The target compounds were separated on a DB-5MS column (30 m×0.25 mm×1 µm). Identification and quantification were achieved using an electron impact (EI) ion source in the positive ion mode with the selected ion monitoring mode. The internal standard method was used to quantify the 3-chloropropanediol esters, 2-chloropropanediol esters, and glycidyl esters. Under the optimal conditions, the correlation coefficients of the standard calibration curves were greater than 0.999 in the mass concentration range of 0.01-0.80 mg/L. The limits of detection were 25, 25, and 20 µg/kg (S/N=3), and the limits of quantification were 75, 75, 60 µg/kg (S/N=10). Four samples of different matrix types were selected for scaling experiments. At spiked levels of 250, 500, and 750 µg/kg, the recoveries of 3-chloropropanediol esters, 2-chloropropanediol esters, and glycidyl esters in spiked samples ranged from 89.0% to 98.7%, with relative standard deviations between 2.05% and 7.81% (n=6). This method was used to determine 112 commercially available vegetable oil samples, among which 84 samples were detected with 3-chloropropanediol esters, 2-chloropropanediol esters, or glycidyl esters. The method developed in this study was remarkably different from the standard method, which are mentioned in the national standard method (GB 5009.191-2016) and industry standard method (SN/T 5220-2019), especially in the pretreatment step that involved acidic transesterification. Use of the acidic transesterification method can avoid side reactions, such as the conversion of 3-chloropropanediol, 2-chloropropanediol, and 3-bromopropanediol to free glycidol under alkaline conditions. The method developed in this study was more efficient, and the results were more accurate and reproducible. It has theoretical and practical significance for the control of 3-chloropropanediol esters, 2-chloropropanediol esters, and glycidyl esters residues in vegetable oils, establishment of detection standards, and optimization of the production process.

[Determination of free amino acids in Eriocheir sinensis by ultra-high performance liquid chromatography-high resolution mass spectrometry].

Eriocheir sinensis is a unique freshwater crab found in China, which is well known for its rich nutrition and sweet and delicious taste. Free amino acids in Eriocheir sinensis are not only important nutrients but also are closely related to their unique taste and aroma. Therefore, the determination of the free amino acid contents in Eriocheir sinensis is of great significance for product quality evaluation, flavor research, authenticity, and origin identification. Herein we proposed an ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS)-based method for the determination of 17 free amino acids in Eriocheir sinensis. First, 5 g of the Eriocheir sinensis sample was weighed into a 50-mL polypropylene centrifuge tube. Then, 10 mL of extraction solvents was added to the centrifuge tube, and the resultant solution was mixed well using a vortex mixer. We compared a variety of solvents and finally selected 5%(v/v) perchloric acid aqueous solution as the optimum extraction solvent. The supernatant was transferred to another polypropylene centrifuge tube after centrifuging at 8000 r/min for 5 min. The extraction procedure was repeated according to the above-mentioned steps, and the extraction solution was combined with the supernatant. The extracts were then adjusted to pH 6.5 with 1 mol/L potassium hydroxide solution, and were diluted to 50 mL with water. After filtering by both qualitative filter paper and a 0.45-µm polyether sulfone syringe filter, the extracts were determined by UHPLC-HRMS. We compared three types of mobile phases and chose 0.1%(v/v) formic acid aqueous solution mixed with acetonitrile as the optimum one. Precise parent ion and ion source parameters were also optimized. The 17 analytes, viz. aspartic acid, threonine, serine, glutamic acid, proline, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, arginine, glycine, alanine, and histidine, were separated on an XDB-C18 column (100 mm×4.6 mm, 1.7 µm) with gradient elution. The amino acids were then detected by HRMS in electrospray ionization and selected ion monitoring modes, and the analytes were quantified using external standards. The instrumental detection limit (IDL) and the instrumental quantification limit (IQL) were 0.3 mg/L and 1.0 mg/L, respectively. The linear correlation coefficients were all above 0.9990 in the concentration range of 10.0-200.0 mg/kg. Three levels of free amino acid standards were spiked into the edible parts of Eriocheir sinensis. The recoveries of the amino acids were between 78.4% and 105.3%. The intra-sample, intra-day, and inter-day repeatabilities were below 4.2%, 5.2%, and 11.4%, respectively, which were within reasonable ranges. Twenty samples of Eriocheir sinensis were tested using the proposed method. Thus, in this study, we developed an alternative method for the determination of free amino acids in Eriocheir sinensis with simple pretreatment, good selectivity, and high accuracy.

[Determination of four trihalomethanes in ship ballast water by gas chromatography-negative chemical ionization-mass spectrometry].

Ship ballast water can control the roll, trim, and draft of the ship, and thus ensuring the balance and stability of the ship in the course of sailing, and playing a vital role in the safe navigation of ships. The annual discharge of ship ballast water is very large in China. About three to five billion cubic meters of ship ballast water is discharged into offshore or inland waters every year. This water contains plankton, pathogens, and their larvae or spores. If not be handled appropriately, this will have a serious impact on the ecological environment of the discharge waters. Ballast water is usually treated by electrolysis before being discharged. Sodium hypochlorite can be generated, which can kill microorganisms; however, the by-products trihalomethanes (THMs) are cytotoxic and biotoxic. Studies have shown that THMs may cause fetal growth retardation, spontaneous abortion, or death. The concentration of THMs in drinking water is closely related to the risk of bladder cancer death. Hence, it is important to establish a method for the determination of THMs in ship ballast water. The four kinds of THMs are chloroform, dichlorobromomethane, chlorodibromomethane, and tribromomethane. At present, ship ballast water is mostly analyzed by gas chromatography (GC) using an electron capture detector (ECD) or by gas chromatography-mass spectrometry (GC-MS). Given the low boiling point of THMs, headspace injection and purge-and-trap can be used. Gas chromatography-negative chemical ionization-mass spectrometry (GC-NCI-MS), was adopted. NCI is a soft ionization technique that shows special response to compounds bearing electronegative elements or groups. THMs contain electronegative chlorine atoms and bromine atoms. Therefore, NCI is a good choice for their analysis. The samples were processed by the headspace injection technique. The NaCl content in 10 mL sample was optimized in headspace injection. The results showed that 3.0 g NaCl was the most suitable dosage. The analytes were separated on a DB-5MS UI capillary-column (30 m×0.25 mm×1.0 µm). The target compounds were quantified by using the external standard method in selected ion monitoring (SIM) mode. The four THMs were not only well separated but also showed a high response at 0.2 µg/L. The four THMs showed good linear relationships in the range of 0.2-50 µg/L, with correlation coefficients≥0.995. The limits of quantification (LOQs, S/N=10) were 0.1-0.2 µg/L, and the average recoveries of the four THMs were 90.3%-106.8% at the three spike levels of 0.2, 0.5, and 2.0 µg/L. The relative standard deviations were 1.4%-6.2%. The LOQs of the THMs in the GB/T 5750.8-2006 Standard Test Method of Drinking Water Organic Matter Index are 0.3-6.0 µg/L. It can be seen that the LOQs of the THMs are greatly reduced in this study. The proposed method is accurate, stable, and reliable, and it can be used for monitoring the four THMs in ship ballast water. The method was applied for the detection of 36 ship ballast water samples. In all cases, the detection rates of tribromomethane, chlorodibromomethane, dichlorobromomethane, and chloroform were 83.3%, 69.4%, 22.2%, and 19.4%, respectively. The detection values of tribromomethane, chlorodibromomethane, dichlorobromomethane, and chloroform were 34.25-221.5 µg/L, 3.52-41.87 µg/L, 1.52-8.56 µg/L, and 0.02-5.46 µg/L, respectively. Based on the analysis of several ship ballast water samples (electrolytic water), it was concluded that the greater the number of bromine atoms in the THMs, the higher are the detection rate and detection value in ship ballast water. Compared to chloroform, tribromomethane is more harmful to living beings. China has acceded to the International Convention on Ship Ballast Water and Sediment Control and Management. There is an urgent need to establish analysis methods with high sensitivity, good stability, and high accuracy in addition to determining standards and regulations for ship ballast water.

A new ionic liquid bridged periodic mesoporous organosilicas stationary phase for per aqueous liquid chromatography and its application in the detection of biogenic amines.

In order to solve the problems of using a large proportion of acetonitrile on the hydrophilic interaction liquid chromatography (HILIC) columns that was not environmentally friendly, and the poor acid and base resistance of traditional bonded silica columns, we reported a novel stationary phase of Au nanoparticles (Au NPs) covalently bonded to ionic liquid (ILs) bridged periodic mesoporous organosilicas (PMO) hydrophilic microspheres (PMO-ILs-Au NPs) for per aqueous liquid chromatography (PALC). The PMO hydrophilic microspheres were prepared by condensation of 1,3-bis(trimethoxysilylpropyl)imidazoliumchloride and 1, 2-Bis (triethoxysilyl) ethane and then modified with Au NPs the surface. The obtained materials were characterized by elemental analysis, FT-IR spectra, scanning electron microscope and transmission electron microscopy. The retention behavior was evaluated by investigating the effect of various chromatographic factors on the retention of different types of solutes. The retention mechanism of the stationary phases in PALC was a mixed type of anion-exchange and hydrophobic interaction. Compared with C18-SiO2 column, the acid and base resistance of the stationary phase were greatly improved. Compared with the HILIC column and C18 column, some hydrophilic compounds such as six organic acids and eight biogenic amines were baseline separated with the enhanced resolution of the PMO-ILs-Au NPs column under the PALC mode. The efficiency of the new column was significantly higher than that of the HILIC column. Furthermore, the analysis of PALC-triple quadrupole mass spectrometry was developed for simultaneous detection of eight biogenic amines. This method could improve detection efficiency, save reagent and reduce environmental pollution. PALC as a green chromatography analytical method was suitable for the replacement of HILIC.

A Mathematical Model of the Mouse Atrial Myocyte With Inter-Atrial Electrophysiological Heterogeneity.

Biophysically detailed mathematical models of cardiac electrophysiology provide an alternative to experimental approaches for investigating possible ionic mechanisms underlying the genesis of electrical action potentials and their propagation through the heart. The aim of this study was to develop a biophysically detailed mathematical model of the action potentials of mouse atrial myocytes, a popular experimental model for elucidating molecular and cellular mechanisms of arrhythmogenesis. Based on experimental data from isolated mouse atrial cardiomyocytes, a set of mathematical equations for describing the biophysical properties of membrane ion channel currents, intracellular Ca2+ handling, and Ca2+-calmodulin activated protein kinase II and ß-adrenergic signaling pathways were developed. Wherever possible, membrane ion channel currents were modeled using Markov chain formalisms, allowing detailed representation of channel kinetics. The model also considered heterogeneous electrophysiological properties between the left and the right atrial cardiomyocytes. The developed model was validated by its ability to reproduce the characteristics of action potentials and Ca2+ transients, matching quantitatively to experimental data. Using the model, the functional roles of four K+ channel currents in atrial action potential were evaluated by channel block simulations, results of which were quantitatively in agreement with existent experimental data. To conclude, this newly developed model of mouse atrial cardiomyocytes provides a powerful tool for investigating possible ion channel mechanisms of atrial electrical activity at the cellular level and can be further used to investigate mechanisms underlying atrial arrhythmogenesis.

The Role of CaMKII Overexpression and Oxidation in Atrial Fibrillation-A Simulation Study.

This simulation study aims to investigate how the Calcium/calmodulin-dependent protein kinase II (CaMKII) overexpression and oxidation would influence the cardiac electrophysiological behavior and its arrhythmogenic mechanism in atria. A new-built CaMKII oxidation module and a refitted CaMKII overexpression module were integrated into a mouse atrial cell model for analyzing cardiac electrophysiological variations in action potential (AP) characteristics and intracellular Ca2+ cycling under different conditions. Simulation results showed that CaMKII overexpression significantly increased the phosphorylation level of its downstream target proteins, resulting in prolonged AP and smaller calcium transient amplitude, and impaired the Ca2+ cycling stability. These effects were exacerbated by extra reactive oxygen species, which oxidized CaMKII and led to continuous high CaMKII activation in both systolic and diastolic phases. Intracellular Ca2+ depletion and sustained delayed afterdepolarizations (DADs) were observed under co-existing CaMKII overexpression and oxidation, which could be effectively reversed by clamping the phosphorylation level of ryanodine receptor (RyR). We also found that the stability of RyR release highly depended on a delicate balance between the level of RyR phosphorylation and sarcoplasmic reticulum Ca2+ concentration, which was closely related to the genesis of DADs. We concluded that the CaMKII overexpression and oxidation have a synergistic role in increasing the activity of CaMKII, and the unstable RyR may be the key downstream target in the CaMKII arrhythmogenic mechanism. Our simulation provides detailed mechanistic insights into the arrhythmogenic effect of CaMKII overexpression and oxidation, which suggests CaMKII as a promising target in the therapy of atrial fibrillation.

[Determination of eight vitamin E in vegetable oils by gas chromatography-mass spectrometry and its application on authentication of sesame oil].

A method was established for the determination of eight vitamins E (α-, ß-, γ-, δ-tocopherol and α-, ß-, γ-, δ-tocotrienol) in vegetable oils using gas chromatography-mass spectrometry (GC-MS). The targets were extracted with methanol, and analyzed by GC-MS in the selected ion monitoring (SIM) mode after concentration to a constant volume, and quantified using the external standard method. Baseline separation were achieved for all the target compounds. The linearities of all the compounds were between 0.01 and 1 mg/L. The limits of detection (LODs) and limits of quantification (LOQs) were in the range of 0.03-0.25 mg/kg and 0.10-0.83 mg/kg, respectively. The average recoveries of all the targets in sesame oil samples were between 87.5% and 107.4% at three spiked levels (10, 50, and 250 mg/kg), and the RSDs were all less than 7.5%. The tocopherols and tocotrienols contents in sesame oil samples and in six lower-price vegetable oils (soybean, rapeseed, sunflower, peanut, corn and palm oils) were determined by the above mentioned method. The results showed that the vitamin E profiles of sesame oil were significantly different from those of the other six vegetable oils. Therefore, vitamin E can be used as a discriminating parameter for detecting the adulteration of sesame.

New versatile zincic sorbent for tobacco specific nitrosamines and lead ion capture.

New carbon-doped ferric zinc oxide sorbents were fabricated to capture the environment carcinogen tobacco specific nitrosamines (TSNA) efficiently in solution, following new adsorption model of electrostatic attraction instead of traditional geometric constraints. The influence of ferric content on the structure-property of the sorbents was systemically studied with XRD, N2 adsorption-desorption and SEM methods combined with the adsorption of TSNA in different solutions. New sorbent captured 99% of 4-methylnitrosamino-1-3-pyridyl-1-butanone (NNK) in simulated surface water and 40% of TSNA in the tobacco extract solution, more than activated carbon or zeolites. Ferric ZnO sorbent took about 15 min to reach the adsorption equilibrium in the NNK or Pb(Ⅱ) solution, faster than NaZSM-5 zeolite. Moreover, the adsorbed NNK on ferric ZnO sorbent decomposed at mild conditions for the first time, providing a new way to control environment pollution.

[Determination of amitraz and its metabolites in vegetables and fruits by high performance liquid chromatography-tandem mass spectrometry].

A method was established for the determination of amitraz (AMZ), and its metabolites semiamitraz (DMPF), 2,4-dimethylformamidine (DMF) and 2,4-dimethylaniline (DMA) in vegetables and fruits by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The samples were diluted with 0.1 mol/L sodium hydroxide and extracted with n-hexane-isopropanol (2:1, v/v). The separation was performed on a Phenomenex Kinetex C18 column (100 mm×4.6 mm, 2.6 µm) with gradient elution using 0.1% (v/v) formic acid aqueous solution-methanol as the mobile phase. The analysis of amitraz and its metabolites were detected under electrospray positive ionization mode. The limits of quantification (LOQs) were between 0.01 and 0.4 µg/kg. The good linearities (r>0.99) were achieved for the target compounds in the range of 1.0-200.0 µg/L. The recoveries at three spiked levels (0.5, 5.0 and 20 µg/kg) in blank matrix were in the range of 62.5%-105.0% with the relative standard deviations between 7.5% and 17.6%. The method is convenient, rapid, accurate, efficient, sensitive and practical. It is suitable for the determination and confirmation of amitraz and its metabolites in vegetables and fruits, and can meet the demands of domestic and foreign regulations.

[Determination of 10 perfluorinated carboxylic acid compounds in water by gas chromatography-mass spectrometry coupled with negative chemical ionization].

A method was developed for the simultaneous determination of 10 perfluorinated carboxylic acid compounds in water by gas chromatography-mass spectrometry coupled with negative chemical ionization (GC-NCI-MS). Perfluorinated carboxylic acid compounds were derivatized by trifluoro-N-methyl-N-(trimethylsilyl) acetamide (MSTFA) as the trimethylsilyl derivatization reagent. The water sample was purified and enriched through a weak anion exchange solid phase extraction column and analyzed via GC-NCI-MS. The sample pretreatment, derivation and instrument conditions were optimized. The results showed that the linearity of the 10 perfluorinated carboxylic acid compounds was good in the range of 0.1-10 mg/L with correlation coefficients of 0.9956-0.9993. The limits of detection (LODs) and limits of quantification (LOQs) were 0.5-1.5 µg/L and 1.5-4.5 µg/L, respectively. The spiked recoveries of the blank samples ranged from 70.2% to 112.6% with the relative standard deviations (RSDs) between 2.1% and 14.5% (n=6). The method is simple, sensitive, accurate and precise, and can be used to detect the 10 perfluorinated carboxylic acid compounds in water.

[Determination of nine pesticide residues in honey using high performance liquid chromatography-tandem mass spectrometry].

A method is proposed for the simultaneous determination of nine benzimidazole and neonicotinoid pesticides present in honey by employing automatic solid-phase extraction with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). A honey sample was dissolved in a phosphate buffer (pH=7.8) followed by ultrasonic extraction. The extracts were then purified through solid-phase extraction (SPE) with hydrophilic-lipophilic balance (HLB) cartridges. Finally, nitrogen was blown on the obtained mixture, and the mixture was subsequently filtered for conducting HPLC-MS/MS analysis. Nine compounds were detected under the multiple reaction monitoring (MRM) mode, and the corresponding quantification was performed by employing the method of internal standards. The nine detected pesticides demonstrated good linearity in the range of 0.002-0.05 mg/L, with the correlation coefficient values (r2) being higher than 0.99. The limits of detection (LODs) (S/N=3) and limits of quantification (LOQs) (S/N=10) were found to be in the ranges of 0.1-1.0 µg/kg and 0.3-2.0 µg/kg, respectively. Furthermore, the results indicated that the recoveries of the nine detected pesticides range from 78.2%-101.2% at three spiked levels of 5.0, 10.0, and 20.0 µg/kg with a relative standard deviation (RSD) range of 1.3%-14.3% (n=6). Hence, the proposed method is rapid and can be employed for accurate determination of pesticide residues in large quantities of honey samples.

[Determination of organo-tin residues in edible vegetable oil by positive chemical ionization-gas chromatography-mass spectrometry].

For the determination of organo-tin residues in edible vegetable oil, a method was developed based on gas chromatography-mass spectrometric (GC-MS) with positive chemical ionization (PCI). The edible oil samples were first dissolved by cyclohexane-ethyl acetate (1:1, v/v), and then purified by gel permeation chromatography (GPC). After derivatization by sodium tetraethylborate, the samples were analyzed by GC-MS with PCI source in the single ion monitor (SIM) mode. The seven organo-tin compounds showed good linear relationships in the range of 20-2000 µg/L and the correlation coefficients exceeded 0.99. The limits of quantitation (LOQs) and the average recoveries of the seven organo-tin compounds were 0.3-1.2 µg/kg and 66.2%-103.2%, respectively, and the relative standard deviations were less than 11.5% at three spike levels (0.05, 0.10, and 0.20 mg/kg). The method showed good linearity and high sensitivity and can be used for the determination of organo-tin residues in edible vegetable oil.