Intelligent analysis of excitation-emission matrix fluorescence fingerprint to identify and quantify adulteration in camellia oil based on machine learning.

Camellia oil (CAO) is a premium edible vegetable oil with medical value and biological activity, but it is susceptible to adulteration. Therefore, the demand for intelligent analysis to decipher the category and proportion of adulterated oil in CAO was the main driver of this work. Excitation-emission matrix fluorescence (EEMF) spectra of 933 vegetable oil samples were characterized by a chemometric method to obtain chemically meaningful information. Authenticity identification models were constructed using four machine learning methods to realize the discrimination of oil species adulterated in CAO mixtures. Meanwhile, quantitative models were established aiming at the fraud of CAO proportion in blended oil. Results showed that the specially constructed CNN obtained the optimal performance when evaluating unseen real-world samples, with a classification accuracy of 95.8% and 92.2%, and mean-absolute quantitative errors between 2.6 and 6.7%. Therefore, EEMF fingerprints coupled with machine learning are expected to provide intelligent and accurate analysis for authenticity detection of CAO.

Chemometrics-enhanced high-performance liquid chromatography-diode array detection strategy to quantify protoberberine alkaloids in varying Coptidis Rhizoma-related medicines.

Quantitation of protoberberine alkaloids is an essential guarantee for efficacy control and medication safety of Coptidis Rhizoma (CR) related medicines. Traditional univariate chromatography faced challenges with co-elution, unknown interferences, and retention time shift when analyzing isomeric analytes in varying sample matrices. We presented a chemometrics-enhanced high-performance liquid chromatography-diode array detection (HPLC-DAD) strategy for simultaneous quantification of six protoberberine alkaloids and processed multi-channels chromatographic-spectral data with four second-order calibration algorithms. Chromatographic conditions were firstly optimized. Four groups of predicted samples were modeled individually with the designed calibration set. Mathematical resolutions were then obtained, and pseudo-univariate regression gave the quantitative concentration of each analyte. Four models were scored on fit, linearity, recovery, and robustness, where alternating trilinear decomposition assisted multivariate curve resolution (ATLD-MCR) exhibited an optimal and stable performance. Besides, the resolved spectra presented high consistency with the actual spectra (r≥0.9993). Limits of quantification (LOQ) fully met the pharmacopoeia stipulation and were 0.17, 0.60, 0.19, 0.74, 0.15, and 0.38 µg mL-1 for columbamine, epiberberine, jatrorrhizine, coptisine, palmatine, and berberine, respectively. The importance of this strategy is to exploit collinearity resolution and additional selectivity that permit accurate quantitation at poor chromatographic resolutions, avoiding individual pretreatment and HPLC optimizations for different samples. This study provides a universal alternative for routine quality assessment of protoberberine alkaloids in CR-related medicines.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry combined with chemometrics to identify the origin of Chinese medicinal materials.

Geographical origin and authenticity are two core factors to promote the development of traditional Chinese medicine (TCM) herbs perception in terms of quality and price. Therefore, they are important to both sellers and consumers. Herein, we propose an efficient, accurate method for discrimination of genuine and non-authentic producing areas of TCM by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Take Atractylodes macrocephala Koidz (AMK) of compositae as an example, the MALDI-TOF MS spectra data of 120 AMK samples aided by principal component analysis-linear discriminant analysis (PCA-LDA), partial least squares discriminant analysis (PLS-DA) and random forest (RF) successfully differentiated Zhejiang province, Anhui province and Hunan province AMK according to their geographical location of origin. The correct classification rates of test set were above 93.3%. Furthermore, 5 recollected AMK samples were used to verify the performance of the classification models. The outcome of this study can be a good resource in building a database for AMK. The combined utility of MALDI-TOF MS and chemometrics is expected to be expanded and applied to the origin traceability of other TCMs.

Candidate Chinese Herbal Medicine Alleviates Methamphetamine Addiction via Regulating Dopaminergic and Serotonergic Pathways.

Methamphetamine (METH) addiction and its induced mental disorders have become a severe worldwide problem. A candidate Chinese herbal medicine (CCHM) in our lab had therapeutic effects on METH-induced locomotor sensitization, however, its chemical and pharmacological profiles remain to be elucidated. The current study aimed to investigate the effect of CCHM on conditioned place preference (CPP) induced by METH and screen the main active ingredients and key targets by using network pharmacology and molecular docking methods. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, Gene ontology (GO) analysis and protein-protein interaction (PPI) network were performed to discover the potential mechanisms. Results showed that CCHM could significantly inhibit METH-induced CPP behaviors in mice. A total of 123 components and 43 targets were screened. According to the network pharmacology analysis, ten hub targets including D(2) dopamine receptor (DRD2) and 5-hydroxytryptamine receptor 3A (HTR3A) were screened. GO analysis and KEGG enrichment indicated that mechanisms of CCHM treatment of METH addiction were related to multiple pathways such as dopaminergic synapse and serotoninergic synapse. Western blot results showed that the protein expressions of DRD2 in nucleus accumbens and prefrontal cortex were significantly decreased in METH group, while the protein expressions of HTR3A were significantly increased. These changes caused by METH could be prevented by CCHM pretreatment. The results of molecular docking displayed that the five active ingredients such as (S)-Scoulerine, Hyndarin, and Beta-Sitosterol had good affinities with DRD2 and HTR3A. In conclusion, this study constructed the CCHM's pharmacologic network for treating METH addiction based on the method of network analysis and experimental verification, and analyzed its major active ingredients and potential targets, indicating a new direction for further revealing its mechanisms of effect on METH addiction.

Terminalia chebula extracts ameliorate methamphetamine-induced memory deficits via activating the ERK and Nrf2 pathway.

As a psychoactive substance abused worldwide, methamphetamine (METH) abuse leads to multiple neurodegenerative symptoms including memory deficits. Terminalia chebula retzius extracts (TREs) isolated by our lab have great antioxidant activity and its effect on METH-induced memory deficits has not been investigated yet. The present study was designed to investigate the protective effect of TREs on METH induced cell apoptosis in vitro and memory deficits in vivo. The results showed that TREs treatment attenuated free radical release and improved cell survival of primary hippocampal neurons after METH injury. In the Morris water maze task, TREs treatment reversed METH-induced learning and memory deficits in acquisition and retention. Moreover, TREs reduced oxidative stress in the serum and hippocampus of mice. Additionally, extracellular regulated protein kinases (ERK1/2) pathway and the nuclear factor E2-related factor 2 (Nrf2) pathway were inactivated after METH treatment, and were significantly activated after TREs pretreatment. These findings suggest that TREs may exert potent neuroprotective effect via activation of both ERK and Nrf2 pathways, thus providing a basis for its potential use for ameliorating memory deficits induced by METH.

Geographical origin traceability of traditional Chinese medicine Atractylodes macrocephala Koidz. by using multi-way fluorescence fingerprint and chemometric methods.

Atractylodes macrocephala Koidz. (AM) is an important plant of traditional Chinese medicine (TCM), and its status can be comparable with ginseng in China. The efficacy and quality of AM are closely related to the place of origin. Hence, we proposed a simple and fast strategy to classify AM from different geographical origins by using multi-way fluorescence fingerprint combined with chemometric methods. AM samples with different dilution levels have different fluorescence characteristics, resulting from different content of fluorescence components and chemical microenvironment. Therefore, AM samples were diluted 5-fold, 10-fold, and 20-fold with 40% ethanol aqueous solution to obtain excitation-emission matrix data, and multi-way (three-way and four-way) data arrays were constructed. And then, the fluorescence fingerprints of AM samples were characterized by three-way and four-way parallel factor analysis (PARAFAC). In addition, four pattern recognition methods were used to classify AM from different provinces. The results show that the four-way data array can provide more abundant information than three-way data arrays, so it is more conducive to sample classification. According to the results obtained from the analysis of four-way data array, the correct classification rate (CCR) of the cross-validation and prediction set obtained by partial least squares-discrimination analysis (PLS-DA) were 90.5% and 100%, respectively. To sum up, the proposed method can be regarded as a powerful, feasible, convenient, reliable, and universal classification tool for the classification of AM samples from different provinces and can be used as a promising method to realize the geographical origin traceability of other TCMs.

The Role of Chinese Herbal Therapy in Methamphetamine Abuse and its Induced Psychiatric Symptoms.

Repeated intake of methamphetamine (METH) leads to drug addiction, the inability to control intake, and strong drug cravings. It is also likely to cause psychiatric impairments, such as cognitive impairment, depression, and anxiety. Because the specific neurobiological mechanisms involved are complex and have not been fully and systematically elucidated, there is no established pharmacotherapy for METH abuse. Studies have found that a variety of Chinese herbal medicines have significant therapeutic effects on neuropsychiatric symptoms and have the advantage of multitarget comprehensive treatment. We conducted a systematic review, from neurobiological mechanisms to candidate Chinese herbal medicines, hoping to provide new perspectives and ideas for the prevention and treatment of METH abuse.

Fast identification of the geographical origin of Gastrodia elata using excitation-emission matrix fluorescence and chemometric methods.

Geographical origin is an important factor affecting the quality of traditional Chinese medicine. In this paper, the identification of geographical origin of Gastrodia elata was performed by using excitation-emission matrix fluorescence and chemometric methods. Firstly, excitation-emission matrix (EEM) fluorescence spectra of Gastrodia elata samples from different geographical origins were obtained. And then three chemometric methods, including multilinear partial least squares discriminant analysis (N-PLS-DA), unfold partial least squares discriminant analysis (U-PLS-DA), and k-nearest neighbor (kNN) method, were applied to build discriminant models. Finally, 45 Gastrodia elata samples could be differentiated from each other by these classification models according to their geographical origins. The results showed that all models obtained good classification results. Compared with the N-PLS-DA and U-PLS-DA, kNN got more accurate and reliable classification results and could identify Gastrodia elata samples from different geographical origins with 100% accuracy on the training and test set. Therefore, the proposed method was available for easily and quickly distinguishing the geographical origin of Gastrodia elata, which can be considered as a promising alternative method for determining the geographic origin of other traditional Chinese medicines.

Cysteiniphilum marinum sp. nov., isolated from coastal seawater.

Six aerobic Gram-negative bacteria were isolated from seawater in Guangdong Province, P.R. China. Cells were observed to be Gram-negative, aerobic, non-motile and non-spore forming. Growth of the designated type strain 19X3-30T occurred at a temperature range of 14-37 °C (optimum, 28 °C), a pH range of 6.0-8.0 (optimum, pH 7) and up to 7.5% NaCl (optimum, 1.5%; w/v), and was enhanced by CO2 and L-cysteine supplementation. The major polar lipids identified in strain 19X3-30T were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The principal cellular fatty acids profile showed the presence of anteiso-C15:0, anteiso-C17:0 and C18:0 (> 8% of total fatty acids), and the respiratory quinone was ubiquinone 8 (UQ-8). According to the analysis of 16S rRNA gene sequences, these strains represented a novel species within the family Fastidiosibacteraceae, sharing maximum similarities with Cysteiniphilum litorale DSM 101832T (96.6%) and Cysteiniphilum halobium DSM 103992T (95.3%). Phylogenetic dendrograms based on 16S rRNA gene and protein marker genes from the genomic sequences both indicated that the strains formed a monophyletic lineage closely linked to the genus Cysteiniphilum, which was also supported by the UPGMA dendrogram based on the MALDI-TOF MS profile. The genomic DNA G + C contents of six strains ranged from 38.0% to 38.1%. Based on different taxonomic genomic metrics, phylogeny and phenotypic features, we propose that the strains warrant the assignment to a novel species, for which the name Cysteiniphilum marinum sp. nov. is proposed. The type strain is 19X3-30T (= KCTC 82154T = CGMCC 1.18585T).

The Neuroprotective Effect of L-Stepholidine on Methamphetamine-Induced Memory Deficits in Mice.

Repeated methamphetamine (METH) exposure can cause severe neurotoxicity to the central nervous system, and lead to memory deficits. L-Stepholidine (L-SPD) is a structurally identified alkaloid extract of the Chinese herb Stephania intermedia, which elicits dopamine (DA) D1-type receptors partial agonistic activity and D2-type receptors antagonistic activity. In this study, we investigated the effect of L-SPD on METH-induced memory deficits in mice and its underlying mechanisms. We found that repeated exposure to METH (10 mg/kg, i.p., once per day for 7 consecutive days) impaired memory functions in the novel object recognition experiment. Pretreatment of L-SPD (10 mg/kg, i.p.) significantly improved METH-induced memory deficits in mice. Meanwhile, the protein expression of dopaminergic D2 receptors in hippocampus area was significantly increased by repeated METH exposure, while the protein expression of dopamine transporter (DAT) was significantly reduced. Additionally, the protein expression of phospho-protein kinase A (p-PKA) was significantly increased by repeated METH exposure. The hyperpolarization-activated cyclic-nucleotide-gated non-selective cation 1 (HCN1) channel, which was a key regulator of memory functions and could be regulated by p-PKA, was also significantly increased by repeated METH exposure. These changes caused by METH could be prevented by L-SPD pretreatment. Therefore, our data firstly showed that pretreatment of L-SPD exhibited the protective effect against METH-induced memory deficits, possibly through reducing METH-induced upregulation of dopaminergic pathway and HCN1 channels.

Rapid identification and quantification of cheaper vegetable oil adulteration in camellia oil by using excitation-emission matrix fluorescence spectroscopy combined with chemometrics.

Camellia oil is a high quality oil mainly produced in southern China. It is common that unscrupulous merchants attempt to make huge profits by adulterating camellia oil with other cheaper or lower-quality vegetable oils. Therefore, this paper proposed excitation-emission matrix fluorescence spectroscopy combined with chemometric methods for the rapid identification and quantification of camellia oil adulteration with other cheaper vegetable oils. A five-component parallel factor analysis (PARAFAC) model roughly completed spectral characterization of oil samples, and obtained chemically meaningful information. Four advanced chemometrics methods were used for the classification of camellia oil and other vegetable oils (model 1) and the classification of camellia oil and adulterated camellia oil (models 2 and 3), respectively. Two-directional two-dimensional linear discriminant analysis ((2D)2LDA) was used for chemical data for the first time and showed huge potential. Furthermore, the developed N-PLS regression model used for the prediction of adulteration level in camellia oil showed satisfactory accuracy.

[Intervention of Catalpol on High-fat Diet Induced Nonalcoholic Fatty Liver Disease in Mice].

Objective To investigate the effect of catalpol on high-fat diet(HFD)-induced nonalcoholic fatty liver disease(NAFLD)and its underlying molecular mechanisms.Methods Sixty C57BL/6J male mice were randomly divided into six groups:control group;HFD group;HFD+catalpol(100 mg/kg)group;HFD+catalpol(200 mg/kg)group;HFD+catalpol(400 mg/kg)group;and HFD+atorvastatin calcium(ATC)(30 mg/kg)group.The control group was fed a normal diet containing 4.4 kJ/g fat,whereas the other five groups were fed a high-fat diet containing 19.8 kJ/g fat.Mice in the catalpol or ATC treatment groups were administered by gavage for different doses of catalpol or ATC,whereas other mice were treated with saline.Body weight was measured once a week.Experiments were terminated after 18 weeks,and blood and liver samples were collected after an overnight fast(12 hours)for analysis.The body weight and liver weight were measured and the levels of serum total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate transaminase(AST)as well as inflammatory factors tumor necrosis factor-α(TNF-α),interleukin(IL)-1ß,and IL-6 were determined by commercially available kits.Liver sections were stained with Oil Red O and HE to investigate the lipid accumulation and histopathological changes.The protein expressions of nuclear factor kappa-B(NF-κB)p65,inhibitor of nuclear factor kappa-B α(IκBα),B-cell lymphoma-2(Bcl-2),Bcl-2 associated x protein(Bax),and Caspase-3 were determined by Western blot.Results Compared to the model group,the body weight gains(all P=0.001),liver index(P=0.008,P=0.001,P=0.001),ALT(P=0.004,P=0.001,P=0.001),and AST(P=0.008,P=0.001,P=0.001)were significantly decreased in catalpol treatment groups,and the serum levels of TC(P=0.005,P=0.001),TG (all P=0.001),and LDL-C(all P=0.001)were also significantly decreased in middle and high dose groups,and the serum level of HDL-C was significantly increased in high group(P=0.009).Moreover,compared to the model group,the degree of liver injury and lipid accumulation were obviously decreased in the catalpol treatment groups according to the pathology.Similarly,the release of inflammatory factors was significantly inhibited by the treatment with catalpol.The results of Western blot showed that the protein levels of NF-κB p65(P=0.014,P=0.001,P=0.001)and Caspase-3(all P=0.001)in the livers of HFD-fed mice were significantly reduced by catalpol treatment.In addition,the protein level of IκBα(P=0.028,P=0.001,P=0.001)and the ratio of Bcl-2/Bax in high dose group(P=0.003)was increased by treatment with catalpol.Conclusion Catalpol can effectively improve the body weight gains,liver index,dyslipidemia,and lipid accumulation in HFD-fed mice and inhibit the release of inflammatory factors and hepatocyte apoptosis,thereby preventing the development of NAFLD induced by HFD.

2 Hz EA Reduces Heroin Withdrawal-Induced Hyperalgesia and Heroin Relapse by Downregulating P2X3 Receptors in DRG Neurons.

Electroacupuncture (EA) has effective analgesic effects. Our previous study demonstrated that the upregulation of P2X3 receptors in the dorsal root ganglia (DRG) might participate in heroin withdrawal-induced hyperalgesia. The aim of this study is to further explore whether 2 Hz EA reduces heroin relapse associated with its analgesic effect and whether P2X3 receptors in the DRG are involved in this process. 2 Hz EA was adopted to treat the heroin SA rats in the present study. Heroin-seeking and pain sensitivity were evaluated. The expression of P2X3 receptors in the DRG was detected. Our results showed that compared with the control group, the reinstatement, thermal hyperalgesia, and mechanical allodynia of the heroin-addicted group were increased significantly. The expression of P2X3 receptors in the DRG was increased markedly. After being treated using 2 Hz EA, reinstatement was reduced, hyperalgesia was decreased, and the upregulated expression of P2X3 receptors in the DRG had decreased significantly compared to that in the heroin-addicted group. Consequently, our results indicated that 2 Hz EA was an effective method for treating heroin-induced hyperalgesia and helping prevent relapse, and the potential mechanism might be related to the downregulation of P2X3 receptor expression in the DRG.

Krill oil protects PC12 cells against methamphetamine-induced neurotoxicity by inhibiting apoptotic response and oxidative stress.

Methamphetamine (METH) exposure can cause severe effects to the nervous system; however, the underlying molecular mechanism of neurotoxicity caused by METH is still unclear. Oxidative stress and apoptosis are linked in the pathophysiology of many neurodegenerative diseases. Krill oil (KO) benefits human health via its strong antioxidant ability. Therefore, we hypothesized that KO supplementation might effectively prevent METH-induced neurotoxicity via the inhibition of apoptotic responses and oxidative damages. In this study, PC12 cells were exposed to both METH (3 mmol/L) and KO (0.1, 0.2, 0.4, 0.8 µg/mL) in vitro for 24 h, and the following parameters were measured to detect apoptosis and oxidative stress responses that were triggered by METH: cell viability, the oxidative enzyme system, NO production, ROS production, apoptosis, mitochondrial membrane potential and protein expression of cleaved caspase-3. The results indicate that KO mitigates the apoptotic response post-METH exposure in PC 12 cells by increasing cell viability, decreasing protein expression of cleaved caspase-3, reducing apoptotic rates, and decreasing dissipation of mitochondrial membrane potential. In addition, the study revealed increases in SOD and GSH activity, and decreases in MDA content, NO and ROS production, suggesting that KO is beneficial in reducing oxidative stress, which may also play a role in the regulation of METH-triggered apoptotic response. Consequently, these data indicate that KO could potentially alleviate METH-induced neurotoxicity via the reduction of apoptotic responses and oxidative damages.

Sensitive fluorescence sensing of T4 polynucleotide kinase activity and inhibition based on DNA/polydopamine nanospheres platform.

5'-Polynucleotide kinase (PNK) is a crucial enzyme that catalyzes the phosphorylation of nucleic acid with 5'-OH termini and this phosphorylation reaction has been involved in many important cellular activities. The evaluation of PNK activity has received an increasing attention due to the significance of PNK. Here, the polydopamine nanospheres (PDANS) could adsorb single-stranded DNA (ssDNA) through π-π stacking or hydrogen bonding between nucleobases and aromatic groups of PDANS, while the interaction between double-stranded DNA (dsDNA) with PDANS was weakened due to the changed conformation. Hence, a novel DNA/PDANS platform was constructed for the sensitive and selective determination of T4 PNK activity based on the preferential binding properties of PDANS for ssDNA over dsDNA and the excellent fluorescence quenching property of PDANS. The dye-labeled dsDNA was phosphorylated by T4 PNK and then digested by λ exonuclease, yielding dye-labeled ssDNA, which would be adsorbed on the surface of the PDANS and the fluorescence was greatly quenched by PDANS. Because of the preferential binding properties of PDANS for ssDNA over dsDNA and the high quenching property of PDANS, the developed DNA/PDANS platform exhibited good analytical performance for T4 PNK sensing in complex biological matrix and applied to screening inhibitors. The proposed DNA/PDANS based platform is promising in developing high-throughput assays for drug screening and clinical diagnostics.

[Discussion on related problems of model species of fungus： Paecilomyces hepialid].

Paecilomyces hepiali is a new species of fungus isolated from a field collection of Ophiocordyceps sinensis from Baima snow mountain, Diqing Tibetan Autonomous Prefecture, Yunnan Province by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. The specimen was identified and named as Paecilomyces hepiali by Qing-Tao Chen, the professor of the Institute of Microbiology, Chinese Academy of Sciences (Paecilomyces hepiali) (2008), who identified a dried culture of living strain 82-2 as the holotype. Until now, the holotype (the voucher specimen) was deposited in the Herbarium of the Institute of Chinese Materia Medica (HICMM), China Academy of Chinese Medical Sciences, Beijing. The P. hepiali neotype designated by the paper "Neotypification of P. hepiali (Hypocreales)" published in TAXON 64 (1) by Yao Yi-Jian et al. in February 2015 is untenable.

Core-Shell-Shell Multifunctional Nanoplatform for Intracellular Tumor-Related mRNAs Imaging and Near-Infrared Light Triggered Photodynamic-Photothermal Synergistic Therapy.

A multifunctional nanoplatform, which generally integrates biosensing, imaging diagnosis, and therapeutic functions into a single nanoconstruct, has great important significance for biomedicine and nanoscience. Here, we developed a core-shell-shell multifunctional polydopamine (PDA) modified upconversion nanoplatform for intracellular tumor-related mRNAs detection and near-infrared (NIR) light triggered photodynamic and photothermal synergistic therapy (PDT-PTT). The nanoplatform was constructed by loading a silica shell on the hydrophobic upconversion nanoparticles (UCNPs) with hydrophilic photosensitizer methylene blue (MB) entrapped in it, and then modifying PDA shells through an in situ self-polymerization process, thus yielding a core-shell-shell nanoconstruct UCNP@SiO2-MB@PDA. By taking advantages of preferential binding properties of PDA for single-stranded DNA over double-stranded DNA and the excellent quenching property of PDA, a UCNP@SiO2-MB@PDA-hairpin DNA (hpDNA) nanoprobe was developed through adsorption of fluorescently labeled hpDNA on PDA shells for sensing intracellular tumor-related mRNAs and discriminating cancer cells from normal cells. In addition, the fluorescence resonance energy transfer from the upconversion fluorescence (UCF) emission at 655 nm of the UCNPs to the photosensitizer MB molecules could be employed for PDT. Moreover, due to the strong NIR absorption and high photothermal conversion efficiency of PDA, the UCF emission at 800 nm of the UCNPs could be used for PTT. We demonstrated that the UCNP@SiO2-MB@PDA irradiated with NIR light had considerable PDT-PTT effect. These results revealed that the developed multifunctional nanoplatform provided promising applications in future oncotherapy by integrating cancer diagnosis and synergistic therapy.

Interference-free spectrofluorometric quantification of aristolochic acid I and aristololactam I in five Chinese herbal medicines using chemical derivatization enhancement and second-order calibration methods.

A rapid interference-free spectrofluorometric method combined with the excitation-emission matrix fluorescence and the second-order calibration methods based on the alternating penalty trilinear decomposition (APTLD) and the self-weighted alternating trilinear decomposition (SWATLD) algorithms, was proposed for the simultaneous determination of nephrotoxic aristolochic acid I (AA-I) and aristololactam I (AL-I) in five Chinese herbal medicines. The method was based on a chemical derivatization that converts the non-fluorescent AA-I to high-fluorescent AL-I, achieving a high sensitive and simultaneous quantification of the analytes. The variables of the derivatization reaction that conducted by using zinc powder in acetose methanol aqueous solution, were studied and optimized for best quantification results of AA-I and AL-I. The satisfactory results of AA-I and AL-I for the spiked recovery assay were achieved with average recoveries in the range of 100.4-103.8% and RMSEPs <0.78ngmL-1, which validate the accuracy and reliability of the proposed method. The contents of AA-I and AL-I in five herbal medicines obtained from the proposed method were also in good accordance with those of the validated LC-MS/MS method. In light of high sensitive fluorescence detection, the limits of detection (LODs) of AA-I and AL-I for the proposed method compare favorably with that of the LC-MS/MS method, with the LODs <0.35 and 0.29ngmL-1, respectively. The proposed strategy based on the APTLD and SWATLD algorithms by virtue of the "second-order advantage", can be considered as an attractive and green alternative for the quantification of AA-I and AL-I in complex herbal medicine matrices without any prior separations and clear-up processes.

Simultaneous determination of umbelliferone and scopoletin in Tibetan medicine Saussurea laniceps and traditional Chinese medicine Radix angelicae pubescentis using excitation-emission matrix fluorescence coupled with second-order calibration method.

A chemometrics-assisted excitation-emission matrix (EEM) fluorescence method is presented for simultaneous determination of umbelliferone and scopoletin in Tibetan medicine Saussurea laniceps (SL) and traditional Chinese medicine Radix angelicae pubescentis (RAP). Using the strategy of combining EEM fluorescence data with second-order calibration method based on the alternating trilinear decomposition (ATLD) algorithm, the simultaneous quantification of umbelliferone and scopoletin in the two different complex systems was achieved successfully, even in the presence of potential interferents. The pretreatment is simple due to the "second-order advantage" and the use of "mathematical separation" instead of awkward "physical or chemical separation". Satisfactory results have been achieved with the limits of detection (LODs) of umbelliferone and scopoletin being 0.06ngmL(-1) and 0.16ngmL(-1), respectively. The average spike recoveries of umbelliferone and scopoletin are 98.8±4.3% and 102.5±3.3%, respectively. Besides, HPLC-DAD method was used to further validate the presented strategy, and t-test indicates that prediction results of the two methods have no significant differences. Satisfactory experimental results imply that our method is fast, low-cost and sensitive when compared with HPLC-DAD method.

Discussion on related problems of model species of fungus： Paecilomyces hepialid / 中国中药杂志

Paecilomyces hepiali is a new species of fungus isolated from a field collection of Ophiocordyceps sinensis from Baima snow mountain, Diqing Tibetan Autonomous Prefecture, Yunnan Province by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. The specimen was identified and named as Paecilomyces hepiali by Qing-Tao Chen, the professor of the Institute of Microbiology, Chinese Academy of Sciences (Paecilomyces hepiali) (2008), who identified a dried culture of living strain 82-2 as the holotype. Until now, the holotype (the voucher specimen) was deposited in the Herbarium of the Institute of Chinese Materia Medica (HICMM), China Academy of Chinese Medical Sciences, Beijing. The P. hepiali neotype designated by the paper "Neotypification of P. hepiali (Hypocreales)" published in TAXON 64 (1) by Yao Yi-Jian et al. in February 2015 is untenable.