Sinomenine increases the methylation level at specific GCG site in mPGES-1 promoter to facilitate its specific inhibitory effect on mPGES-1.

Prostaglandin E2 (PGE2) in cancer and inflammatory diseases is a key mediator of disease progression. Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to inhibit the expression of PGE2 by depressing cyclooxygenase (COX) in inflammatory treatments. However, the inhibition to COXs may cause serious side effects. Thus, it is urgent to develop new anti-inflammatory drugs aiming new targets to inhibit PGE2 production. Microsomal prostaglandin E synthase 1 (mPGES-1) catalyzes the final step of PGE2 biosynthesis. Therefore, the selective inhibition of mPGES-1 has become a promising strategy in the treatments of cancer and inflammatory diseases. Our previous studies confirmed that sinomenine (SIN) is a specific mPGES-1 inhibitor. However, the exact mechanism by which SIN inhibits mPGES-1 remains unknown. This study aimed to explain the regulation effect of SIN to mPGES-1 gene expression by its DNA methylation induction effect. We found that the demethylating agent 5-azacytidine (5-AzaC) reversed the inhibitory effect of SIN to mPGES-1. Besides, SIN selectively increased the methylation level of the promoter region in the mPGES-1 gene while the pretreatment of 5-AzaC suppressed this effect. The results also shows that pretreatment with SIN increased the methylation level of specific GCG sites in the promoter region of mPGES-1. This specific methylation site may become a new biomarker for predicting and diagnosing RA and cancer with high expression of mPGES-1. Also, our research provides new ideas and solutions for clinical diagnosis and treatment of diseases related to mPGES-1 and for targeted methylation strategy in drug development.

[Molecular mechanism of Spatholobi Caulis in treatment of ovarian cancer based on network pharmacology and experimental verification].

The present study explored the main active ingredients and the underlying mechanism of Spatholobi Caulisin the treatment of ovarian cancer(OC) by network pharmacology, molecular docking, and in vitro cell experiments. The active ingredients and their predicted targets(AITs) were first acquired online with the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Theoretical disease targets(DTs) were obtained through professional databases including GeneCards, OMIM, PharmGkb, TTD, and DrugBank. The common targets in the intersection of AITs and DTs were used for the construction of a "drug-ingredient-disease-target" network by Cytoscape 3.7.1. STRING database was used to construct a protein-protein interaction(PPI) network. R 4.0.5 was used for GO and KEGG functional enrichment analyses. Schr9 dinger Maestro was used to perform and optimize the molecular docking and virtual screening.Twenty-three active ingredients of Spatholobi Caulis were screened out, involving 75 OC targets and 178 signaling pathways.Network analysis revealed that Spatholobi Caulis presumedly exerted an anti-OC effect by acting on key protein targets such as GSK-3ß, Bcl-2, and Bax. Molecular docking showed that GSK-3ß possessed goodbinding activity to prunetin. In vitro cell experiments preliminarily verified the core targets and pathways of prunetin, the active ingredient of Spatholobi Caulis against human OC SKOV3 cells.CCK-8 assay was used to detect the cell proliferation, and flow cytometry was used to detect the effect of prunetin on apoptosis of human OC SKOV3 cells.The expression of prunetin targets and related regulatory proteins was detected by Western blot.In vitro cell experiments demonstrated that prunetindisplayed significant inhibitory effects on the proliferation of OC cells and could induce apoptosis of SKOV3 cells. Western blot showed that prunetin could induce SKOV3 cell apoptosis by inhibiting GSK-3ß phosphorylation and regulating the expression of downstream Bcl-2 and Bax proteins. This study reveals the scientific nature of network pharmacology in the prediction and guidance of experimental design, confirming that prunetin can treat OC by blocking the GSK-3ß/Bcl-2/Bax cell signal transduction pathway. The findings are expected to provide a basis for the investigation of the mechanism of Spatholobi Caulis in the treatment of OC.

Systematic investigation on the distribution of four hidden toxic Aconitum alkaloids in commonly used Aconitum herbs and their acute toxicity.

Yunaconitine (YAC), crassicauline A (CCA), 8-deacetylyunaconitine (DYA), and 8-deacetylcrassicauline A (DCA), as hidden toxic Aconitum alkaloids, are detected in some products of processed Aconitum carmichaelii lateral root and poisoning cases. The distribution and toxicity of these four components in Aconitum herbs should be further systematically studied for medication safety. This study developed a new UHPLC-QQQ-MS/MS method to determine ten Aconitum alkaloids, including aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine, benzoylhypaconine, YAC, CCA, DYA, and DCA, for Aconitum herbs simultaneously. YAC and CCA were founded in some samples of unprocessed A. carmichaelii lateral root (7.04%), A. carmichaelii root (9.43%), A. brachypodum root (6.00%), and A. ouvrardianum root (100%). Four hidden toxic Aconitum alkaloids were detected in processed A. carmichaelii lateral root (2.56%) and A. vilmorinianum root (100%). Four hidden toxic Aconitum alkaloids played significant roles in the classification of Aconitum herbs by OPLS-DA analysis. The acute toxicity test was performed by up-and-down procedure (UDP). The oral administration of the half lethal dose (LD50) of YAC, CCA, DYA, and DCA to female ICR mice was 2.37 mg/kg, 5.60 mg/kg, 60.0 mg/kg, and 753 mg/kg, respectively. The LD50 by intravenous injection was 0.200 mg/kg, 0.980 mg/kg, 7.60 mg/kg, and 34.0 mg/kg, respectively. The LD50 of unprocessed A. carmichaelii lateral root, A. vilmorinianum root, and A. brachypodum root to mice orally was 1.89 g/kg, 0.950 g/kg, and 0.380 g/kg, respectively. Symptoms of Aconitum alkaloid poisoning in mice were decreased activity, fur erect, palpebral edema, vomiting, polypnea, and convulsions. The main change of organs was flatulence. No poisoning or death occurred in mice at the maximum dosage (27.0 g/kg) of A. ouvrardianum root orally. To better control the quality and safety of Aconitum herbs, this study provides favorable support for improving the existing standards to strengthen the supervision of the four hidden toxic Aconitum alkaloids.

Potential propensity of traditional herbal materials ingested by collegiate athletes in South China for their consuming health care.

ABSTRACT: In south China, traditional herbal medicines have been widely used as functional foods or dietary supplements for daily health care. Many plant-derived chemical substances with biological activity are inadvertently ingested by collegiate athletes daily through canton-style herbal tea or herbal slow-cooked soup. In the view of the complexity of herbal ingredients, it is still no full survey reported for the sports risk of plant-derived sports doping. This research is firstly a descriptive statistical analysis. Collegiate athletes with different socio-economic characteristics from medical colleges in 3 different regions in China participated in the questionnaire survey. Three survey forms, including the oral interview, email inquiry, handing out and recovering the questionnaires in live, were developed and performed by researchers. It was first found that collegiate athletes resorted to some traditional herbal materials to protect their health care that there were regional differences (P < .01). Collegiate athletes with Health Fitness and Traditional Wushu as their sports expertise showed a higher frequency of recognition or ingestion in the use of traditional herbal materials (P < .01), while their different living types and cuisine preferences did not seem to be associated with the ingestion frequency of traditional herbal materials. In addition, in the view of the significant differences in the use of herbal preparations to relieve sports stress among young athletes in different regions (P < .01), the findings strongly suggested that athletes should strictly control their use of various herbal preparations during sports training and competition, including herbal wines, herbal oils, topical plasters, analgesic tablets.

Identification of blood-red color formation in edible bird's nests provides a new strategy for safety control.

In order to reveal the color formation mechanism of blood-red edible bird's nests (EBNs) and develop a quick and specific strategy to distinguish the artificial fake one, multiple methods of UPLC-TOF/MS, UV, NMR, FT-IR and 2D IR were used to detect the chemical markers of the reddening reaction, the results showed that the reddening substances were C9H10N2O5 and C9H9NO6, which were verified as products of a phenol-keto tautomerism evolved from l-tyrosine. Moreover, natural and artificial red EBNs with varying degrees of chemical fumigation also can be successfully distinguished using the chemical markers, and the protein variation in SDS-PAGE gel could also support the distinction. This work established a systematic method of chemical identification for both natural and artificial blood-red EBNs, and provided a new identification strategy for food safety control that can promote the development of a healthier market of EBNs.

Review on the Development and Applications of Medicinal Plant Genomes.

With the development of sequencing technology, the research on medicinal plants is no longer limited to the aspects of chemistry, pharmacology, and pharmacodynamics, but reveals them from the genetic level. As the price of next-generation sequencing technology becomes affordable, and the long-read sequencing technology is established, the medicinal plant genomes with large sizes have been sequenced and assembled more easily. Although the review of plant genomes has been reported several times, there is no review giving a systematic and comprehensive introduction about the development and application of medicinal plant genomes that have been reported until now. Here, we provide a historical perspective on the current situation of genomes in medicinal plant biology, highlight the use of the rapidly developing sequencing technologies, and conduct a comprehensive summary on how the genomes apply to solve the practical problems in medicinal plants, like genomics-assisted herb breeding, evolution history revelation, herbal synthetic biology study, and geoherbal research, which are important for effective utilization, rational use and sustainable protection of medicinal plants.

Novel SNP markers on ginsenosides biosynthesis functional gene for authentication of ginseng herbs and commercial products.

Panax ginseng and Panax quinquefolius have similar bioactive components and morphological characteristics, but they are known to have different medicinal values, high-sensitive and accurate method is expected to identify the sources of ginseng products and evaluate the quality, but with a huge challenge. Our established UHPLC-TOF/MS method coupled with orthogonal partial least squares discriminant analysis (OPLS-DA) model based on 18 ginsenosides was applied to discriminate the sources of raw medicinal materials in ginseng products, and nested PCR strategy was used to discover 6 novel single nucleotide polymorphism (SNP) sites in functional dammarenediol synthase (DS) gene for genetic authentication of P. ginseng and P. quinquefolius for the first time. OPLS-DA model could identify the sources of raw ginseng materials are real or not. SNP markers were applied to identify ginseng fresh samples as well as commercial products, and proved to be successful. This established molecular method can tell exact source information of adulterants, and it was highly sensitive and specific even when total DNA amount was only 0.1 ng and the adulteration was as low as 1%. Therefore, this study made an attempt at the exploration of new type SNP marker for variety authentication and function regulation at the same time, and the combination of chemical and molecular discrimination methods provided the comprehensive evaluation and authentication for the sources of ginseng herbs and products.

External application of two unrestricted herbal medicines to treat costochondritis in a young collegiate athlete: A case report.

Costochondritis (ChC), especially chronic ChC, typically manifests as spontaneous vague pain in anterior chest area and often occurs in adolescents for unknown reasons; it has prevented many collegiate athletes from participating in physical training and competitions. A 21-year-old female collegiate taekwondo athlete suffering from chronic chest pain was sent by her coaches for diagnosis and treatment. Seated motion palpation was used to identify spontaneous and motion-involved pain areas. Palpation in the supine position was used to initially rule out breast diseases. X-ray, electrocardiogram, and cardiac Doppler ultrasound were used in conjunction with myocardial enzyme testing to rule out lung and cardiovascular diseases. The patient was treated using herbal medicines applied via an external patch. The medicine was comprised of Rhizoma Corydalis and borneol, and the treatment lasted for seven weeks. For five weeks patches were applied at a frequency of two or three times per day, followed by a two-week period of once per day. The patient reported that the pain was relieved after two weeks of external herb use, and the autonomic chest pain had resolved. Re-examination after one month showed that her upper limb range of motion was close to normal, and her psychological burden had almost disappeared. It is possible to seek more active medicinal treatment and more practical external products for young athletes who is suffering chronic ChC that affects the sport training and competitive performances.

Isovaleroylbinankadsurin A ameliorates cardiac ischemia/reperfusion injury through activating GR dependent RISK signaling.

Ischemia/reperfusion (I/R) injury is a pathological process caused by reperfusion. The prevention of I/R injury is of great importance as it would enhance the efficacy of myocardial infarction treatment in patients. Isovaleroylbinankadsurin A (ISBA) has been demonstrated to possess multiple bioactivities for treating diseases. However, its protective effect on myocardial I/R injury remains unknown. In this study, the cardiomyocytes hypoxia/reoxygenation (H/R) in vitro model and coronary artery ligation in vivo model were used to examine the protective effect of ISBA. Apoptosis was determined by flow cytometry and Caspase 3 activity. Protein level was determined by Western blot. The mitochondrial viability was examined with mitochondrial viability stain assay. Mitochondrial membrane potential was detected by JC-1 staining and reactive oxygen species (ROS) was stained with 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA). The binding interactions between ISBA and receptors was simulated by molecular docking. Results showed that ISBA effectively protected cardiomyocytes from I/R injury in in vitro and in vivo models. It remarkably blocked the apoptosis induced by H/R injury through the mitochondrial dependent pathway. Activation of the reperfusion injury salvage kinase (RISK) pathway was demonstrated to be essential for ISBA to exert its protective effect on cardiomyocytes. Moreover, molecular docking indicated that ISBA could directly bind to glucocorticoid receptor (GR) and thus induce its activation. Furthermore, the treatment of GR inhibitor RU486 partially counteracted the protective effect of ISBA on cardiomyocytes, consistent with the results of docking.Most attractively, by activating GR dependent RISK pathway, ISBA significantly elevated the cellular anti-oxidative capacity and hence alleviated oxidative damage induced by I/R injury. In conclusion, our study proved that ISBA protected the heart from myocardial I/R injury through activating GR dependent RISK pathway and consequently inhibiting the ROS generation. It provides a valuable reference for ISBA to be developed as a candidate drug for cardiovascular diseases.

Discovery of chemical markers for identifying species, growth mode and production area of Astragali Radix by using ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry.

BACKGROUND: Astragali Radix (AR) is a well-known Chinese herbal medicine. The quality of AR can be affected by many factors such as species, growth mode and production area, but there are still no chemical markers to distinguish it. PURPOSE: To explore chemical markers for improving the quality assessment of AR and discover chemical markers for identifying species, growth mode and production area of AR. METHODS: A highly sensitive, efficient and accurate method based on ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) for simultaneous quantitative determination of 14 major chemical components (five flavonoids and nine triterpene saponins) in 94 batches of AR from China, Republic of Korea and Germany was developed for the first time. To explore chemical markers and assess changes in the contents of 14 compounds in the 94 batches of AR samples from different regions, hierarchical clustering analysis (HCA) and principal component analysis (PCA) were performed. RESULTS: Astragaloside III was not only an important chemical marker for distinguishing two species of AR, i.e.: Astragalus mongholicus and A. membranaceus, but also a potential chemical marker for the classification of cultivated and semi-wild AR. In addition, in the batches of cultivated AR, the content of isoastragaloside II and cyclocephaloside II were greater in batches from the region of Shaanxi Province than that of other Provinces in China, but the content of calycosin-7-O-ß-D-glucoside and astragaloside IV, which are the quality control markers of AR required by the Chinese Pharmacopoeia, were higher than that of other Provinces in China. In addition, the content of calycosin-7-O-ß-D-glucoside, ononin, calycosin and astragaloside I could be used to identify samples of AR collected from China, Republic of Korea and Germany. CONCLUSION: This UHPLC-QQQ-MS/MS method could be applied to the quantitative evaluation of AR and could be an important and meaningful reference to develop chemical markers for quality control of AR.

[Research on hemostatic effect and mechanism of carbonized Scutellariae Radix on rats of uterine bleeding].

The aim of this paper was to investigate the hemostatic effect and mechanism of carbonized Scutellariae Radix on uterine bleeding in the rats caused by early pregnancy termination. Eight unpregnant female rats were selected as normal group. Forty female rats conceived on the same day received mifepristone(11. 4 mg·kg-1) and misoprostol(125 µg·kg-1) to induce model of incomplete abortion in early pregnancy. Abortion models were randomly divided into model group,carbonized Scutellariae Radix water extract low dose group(0. 55 g·kg-1),medium group(1. 10 g·kg-1),high dose group(2. 20 g·kg-1) and positive control group(0. 45 g·kg-1).The uterine bleeding volume was detected by ultraviolet spectrophotometry. The pathological changes of endometrium were detected by HE(hematixylin-eosin) staining. The levels of interleukin(IL-1ß),IL-6 and tumor necrosis factor(TNF-α) in the plasma of rats were determined by ELISA. The expression levels of IL-1ß,IL-6 and TNF-α mRNA in the uterus of rats were determined by RT-PCR.The protein expression levels of VEGF,MMP-2 and MMP-9 were determined by Western blot. As compared with the normal group,the uterine bleeding volume and histopathological score were increased significantly; microvessel density of endometrial tissues was decreased significantly; the contents of TNF-α,IL-1ß and the levels of TNF-α mRNA and IL-1ß mRNA in the plasma were increased,while the content of IL-6 and level of IL-6 mRNA were decreased significantly. The protein expression levels of VEGF,MMP-2 and MMP-9 in the uterine tissues were also decreased. As compared with the model group,the uterine bleeding volume was decreased significantly in the carbonized Scutellariae Radix medium dose and high dose groups; endometrial repair was promoted,and the microvessel density of endometrial tissues was increased significantly; the contents of IL-1ß and TNF-α in the plasma of rats were decreased significantly,while the content of IL-6 in the plasma of rats was increased significantly; the expression levels of IL-1ß and TNF-α mRNA in the uterus of rats were decreased and the expression level of IL-6 mRNA showed an increase; the protein expressions of VEGF,MMP-2 and MMP-9 were decreased significantly in carbonized Scutellariae Radix medium and high dose groups. In conclusion,carbonized Scutellariae Radix showed good hemostatic effect,and its mechanism may be related to the repair of endometrium and inhibition of inflammatory reaction.

Discovery of the mechanisms and major bioactive compounds responsible for the protective effects of Gualou Xiebai Decoction on coronary heart disease by network pharmacology analysis.

BACKGROUND: Gualou Xiebai decoction (GLXB), a multi-component herbal formula, has been widely used to treat coronary heart disease (CHD) in China for centuries. Several studies have revealed part of its pharmacological activities, whereas its active compounds and mechanisms of action are still unknown because of its complex composition. PURPOSE: Discover the major active compounds and the pharmacological mechanisms of GLXB by network pharmacology methods. METHODS: The main candidate target network was constructed by predicting targets of absorbable chemical compounds of GLXB, collecting therapeutic targets of cardiovascular drugs, constructing target network and layers of screening. Community detection and edge-betweenness calculation were applied to analyze the main candidate target network. Cell viability test, Western blot and flow cytometry were performed to validate the predicted results in cardiomyocytes hypoxia/reoxygenation model. RESULTS: Five clusters and eight cross-talk targets were found in the main candidate target network. Their functions combined together might explain the multifunctional role of GLXB against CHD. Among the cross-talk targets, ESR1 (Estrogen receptor alpha, ERα) and MAPK14 (Mitogen-activated protein kinase 14, p38) were both drug targets and therapeutic targets whose interaction exhibited the greatest edge-betweenness value, suggesting their crucial role in the protective effect of GLXB. The compounds targeting on ESR1 and MAPK14 were identified as apigenin and 25S-macrostemonoside P respectively which were regard as the major bioactive compounds. The predicted results including the major bioactive compounds, their targets and the synergic effects between them were validated. CONCLUSION: This study screened out major bioactive compounds from GLXB and offered a new understanding of the protection mechanism of GLXB against CHD by network pharmacology method and provides a combination strategy to explore mechanisms of action of multi-component drugs from a holistic perspective.

Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis.

To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield, it is important to identify molecular fingerprint of dominant O. sinensis. In the present study, we collected 3 batches of industrially cultivated O. sinensis product with higher yield than the others and compared their internal transcribed spacer (ITS) sequences with the wild and the reported. The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O. sinensis collected. The ITS sequences of O. sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship. The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples. The results showed that all wild and cultivated samples were identified as O. sinensis and all sequences were divided into seven phylogenetic groups in the tree. Those groups were precisely distributed on the map and the process of their system evolution was clearly presented. The three cultivated samples were clustered into two dominant groups, showing the correlation between the industrially cultivated samples and the dominant wild samples, which can provide references for its optimized breeding in the future.

Activation of Nrf2/HO-1 Pathway by Nardochinoid C Inhibits Inflammation and Oxidative Stress in Lipopolysaccharide-Stimulated Macrophages.

The roots and rhizomes of Nardostachys chinensis have neuroprotection and cardiovascular protection effects. However, the specific mechanism of N. chinensis is not yet clear. Nardochinoid C (DC) is a new compound with new skeleton isolated from N. chinensis and this study for the first time explored the anti-inflammatory and anti-oxidant effect of DC. The results showed that DC significantly reduced the release of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharide (LPS)-activated RAW264.7 cells. The expression of pro-inflammatory proteins including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were also obviously inhibited by DC in LPS-activated RAW264.7 cells. Besides, the production of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were also remarkably inhibited by DC in LPS-activated RAW264.7 cells. DC also suppressed inflammation indicators including COX-2, PGE2, TNF-α, and IL-6 in LPS-stimulated THP-1 macrophages. Furthermore, DC inhibited the macrophage M1 phenotype and the production of reactive oxygen species (ROS) in LPS-activated RAW264.7 cells. Mechanism studies showed that DC mainly activated nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, increased the level of anti-oxidant protein heme oxygenase-1 (HO-1) and thus produced the anti-inflammatory and anti-oxidant effects, which were abolished by Nrf2 siRNA and HO-1 inhibitor. These findings suggested that DC could be a new Nrf2 activator for the treatment and prevention of diseases related to inflammation and oxidative stress.

A preliminary study of the effects of medical exercise Wuqinxi on indicators of skin temperature, muscle coordination, and physical quality.

BACKGROUND: Wuqinxi is a traditional medicinal exercise that is widely practiced in China now. Because of its obvious medical rehabilitation, Wuqinxi has been used in the physical education for more than 1.2 million people in at least 24 Chinese Medicine university campuses in China for many years. This investigation aimed to evaluate whether Wuqinxi has the positive effect on physical improvements for female college students. METHODS: Infrared scanners were used for real-time monitoring of body calorie dynamics; the electromyography (EMG) was used to detect the iEMG on biceps, brachioradialis, quadriceps, and gastrocnemius; beside, the physical health elements, heart rate and cardiopulmonary function were also taken within the scope of our investigation and records. RESULTS: Wuqinxi exercise can improve the body function through making the abdominal muscles, back muscles and limbs strength exercise more effectively; Wuqinxi exercise had also made the athletes better control their muscles to have a good way to contraction and keeping balance; Moreover, the performances of speed of 800 m run, setting flexion, set-ups and grip strength had a comprehensive promotion for each of the participants including long-time practitioners and short-term practitioners. CONCLUSIONS: Therefore, the new gymnastics derived from ancient Chinese Wuqinxi exercise can improve the physical health of female college students so that it can be used as part of the development of higher education's health quality in the future.

Specific inhibitor of Notch­3 enhances the sensitivity of NSCLC cells to gemcitabine.

Notch­3 is a receptor of the Notch signaling pathway and plays an important role in regulating self­renewal, differentiation and apoptosis in cancer cells. Overexpression of Notch­3 has been proved to be associated with resistance to gemcitabine (GEM) and poor patient prognosis for various malignant tumors. In the present study, two non­small cell lung cancer (NSCLC) cell lines, H1299 and A549, were induced with GEM for two months and then were treated with various concentrations of a Notch signaling blocker, N­[N­(3,5­difluorophenacetyl)­L­alanyl]­S­phenylglycine t­butyl ester (DAPT), with the goal of reducing expression of Notch intracellular domain 3 (NICD3). Both cell lines were subsequently treated with either DAPT or DAPT combined with GEM and then viability, apoptosis, colony formation and cell count assays were performed. DAPT treatment effectively downregulated the expression of NICD3 in both cell lines. DAPT combined with GEM also significantly reduced the percentage of viable cells in both cell lines, while increasing the percentage of apoptotic cells, compared with GEM alone. In the clonogenicity assays, the combination of DAPT and GEM led to a decrease in clone numbers and significantly greater inhibition of the H1299 and A549 cells compared to treatment with DAPT or GEM alone. Meanwhile, levels of the apoptosis­related proteins, Bcl­2 and Bax, were found to be affected by the various treatments. Thus Notch­3 appears to be a promising target for gene therapy and DAPT is able to mediate a strong antitumor effect in NSCLC cells that overexpress Notch­3. Further studies of a combined treatment regimen with DAPT and GEM are warranted and may provide greater efficacy and safety in the treatment of NSCLC patients.

Simultaneous quantification of nine aconitum alkaloids in Aconiti Lateralis Radix Praeparata and related products using UHPLC-QQQ-MS/MS.

Aconiti Lateralis Radix Praeparata (Fuzi) is obtained from processed daughter roots of Aconitum carmichaeli, a toxic plant with a high medical value well known in Chinese medicine. In addition to the known toxic alkaloids (aconitine, mesaconitine, and hypaconitine) and bioactive alkaloids (benzoylaconine, benzoylmesaconine, and benzoylhypaconine), three rarely found alkaloids have been previously reported in Fuzi, i.e., yunaconitine, 8-deacetyl-yunaconitine, and crassicauline A, and they were reported in recent years to cause potential risk to patients who took Fuzi or related products. To better control the quality of this herb and its related products and ensure safe use, developing a method to simultaneously determine these 9 alkaloids is important. In this research, sensitive and accurate ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry method was established and used to examine 51 Fuzi and 27 Fuzi-containing products. Unexpectedly, 8-deacetyl-yunaconitine was detected in 17 Fuzi samples (33.3%) and 3 Fuzi-containing products (11.1%); yunaconitine in 10 Fuzi samples (19.6%) and 10 Fuzi-containing products (37.0%); and crassicauline A in 3 Fuzi samples (5.8%). Industry and clinics should be aware of the unusually high detection rate of these three toxic alkaloids in the Fuzi herb and its related products and take the necessary precautions to protect patients from any potential risk.

A new approach for identification of medicinal almonds by fourier transform infrared spectroscopy and systematic clustering of characteristic peaks.

Medicinal almonds have been used for over 2 000 years and its clinical efficacy includes relieving cough and asthma. The domestic market in China is flooded with different kinds of dried almonds, such as bitter almond (Armeniacae Semen Amarum, AAS), sweet almond (Armeniacae Semen Dulce, ADS), salted almond (Armeniacae Semen Salsa, ASS), and their sulfur-fumigating products (Armeniacae Semen Sulphur Fumabat, ASFS). Wide varieties of almonds may lead to uncertain efficacy, aberrant quality, and even increased safety risk. However, the authentication method for medicinal almonds has not been reported, although imposters may lead to ineffective medical response. In the present study, Fourier transform infrared spectroscopy (FTIR) and the 2-dimensional infrared (2D-IR) spectroscopy were used to identify different almonds, which were extracted with different solvents including water, methanol, ethanol, chloroform and ethyl acetate, respectively. A new simple FTIR method was developed in the present study. According to the gradient solvent polarity, a new 2D IR method was first developed, and the commodities of almonds in China were analyzed by using the FTIR spectroscopy supported by hierarchical clustering of characteristic peaks. Moreover, 5-hydroxymethyl-2-furfural could be used as a detection index and control target in the quality control of medicinal almonds.

Suppressing mPGES-1 expression by sinomenine ameliorates inflammation and arthritis.

Recently, microsomal prostaglandin E synthase 1 (mPGES-1) has attracted much attention from pharmacologists as a promising strategy and an attractive target for treating various types of diseases including rheumatoid arthritis (RA), which could preserve the anti-inflammatory effect while reducing the adverse effects often occur during administration of non-steroidal anti-inflammatory drugs (NSAIDs). Here, we report that sinomenine (SIN) decreased prostaglandin (PG)E2 levels without affecting prostacyclin (PG)I2 and thromboxane (TX)A2 synthesis via selective inhibiting mPGES-1 expression, a possible reason of low risk of cardiovascular event compared with NSAIDs. In addition, mPGES-1 protein expression was down-regulated by SIN treatment in the inflamed paw tissues both in carrageenan-induced edema model in rats and the collagen-II induced arthritis (CIA) model in DBA mice. More interestingly, SIN suppressed the last step of mPGES-1 gene expression by decreasing the DNA binding ability of NF-κB, paving a new way for drug discovery.

[Uses of fresh herbs with antioxidant effect and prospect for population ecology reconstruction in Macau].

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.