Lysidrhodosides A-I, acylphloroglucinol glucosides with anti-inflammatory activities from the leaves of Lysidice rhodostegia.

Lysidrhodosides A-I (1-9), nine acylphloroglucinol glucoside derivatives along with three known analogues (10-12) were isolated from the leaves of Lysidice rhodostegia. Their structures and absolute configuration were elucidated by spectroscopic data analysis (NMR, UV, IR, HR-ESI-MS), single-crystal X-ray diffraction, and acid hydrolysis with HPLC analysis. Notably, compounds 7-9 represent the first examples of 3-methylbutyryl phloroglucinol glucoside dimers isolated from this plant. Additionally, compounds 1-12 were assessed for their inhibitory effects on nitric oxide (NO) in the LPS-induced BV-2 cells. The results showed that compounds 6 and 12 significantly inhibited the production of the inflammatory mediator NO, with an inhibitory rate of 95.96 and 91.13% at a concentration of 50 µM, respectively.

Anti-inflammatory effect of dictamnine on allergic rhinitis via suppression of the LYN kinase-mediated molecular signaling pathway during mast cell activation.

Mast cells (MCs) are important therapeutic targets for allergic diseases. High-affinity immunoglobulin E (IgE) Fc receptors (FcεRI) trigger abnormal activation of MCs. Allergic rhinitis (AR) is an IgE-mediated antigen inhalation reaction that occurs in the nasal mucosa. MC aggravation and dysfunction were observed during the early stages of AR pathogenesis. Herb-derived dictamnine exhibits anti-inflammatory effects. Here, we investigated the pharmacological effects of herb-derived dictamnine on IgE-induced activation of MCs and an ovalbumin (OVA)-induced murine AR model. The results indicated that dictamnine attenuated OVA-induced local allergic reactions and reduced body temperature in OVA-challenged mice with active systemic anaphylaxis. Additionally, dictamnine decreased the frequency of nasal rubbing and sneezing in an OVA-induced murine AR model. Moreover, dictamnine inhibited FcεRI-activated MC activation in a dose-dependent manner without causing cytotoxicity, reduced the activation of the tyrosine kinase LYN in LAD2 cells, and downregulated the phosphorylation of PLCγ1, IP3R, PKC, Erk1/2, and Akt, which are downstream of LYN. In conclusion, dictamnine suppressed the OVA-stimulated murine model of AR and activated IgE-induced MCs via the LYN kinase-mediated molecular signaling pathway, suggesting that dictamnine may be a promising treatment for AR.

Study on mechanism of hepatoprotective effect of Chrysanthemum morifolium Ramat. based on metabolomics with network analysis and network pharmacology.

Hangju (HJ), the dried flower heads of Chrysanthemum morifolium Ramat., has a significant hepatoprotective effect. However, its underlying protection mechanism against acute liver injury (ALI) has been unclear. An integrated strategy based on metabolomics with network analysis and network pharmacology was developed to explore the potential molecular mechanism of HJ on ALI protection. Firstly, differential endogenous metabolites were screened and identified by metabolomics approach and metabolic pathway analysis was performed by MetaboAnalyst. Secondly, marker metabolites were used to construct metabolite-response-enzyme-gene networks and discover hub metabolites and potential gene targets in network analysis. Thirdly, hub genes through the protein-protein interaction (PPI) network were acquired by the aid of network pharmacology. Finally, the gene targets were taken to intersect with the relevant active ingredients for validation by molecular docking. In total, 48 flavonoids were identified in HJ, which were associated with 8 potential therapeutic targets in network pharmacological analysis. Biochemistry and histopathology analysis demonstrated that HJ exerted hepatoprotective effects. 28 biomarkers were successfully identified as possible biomarkers for the prevention of ALI. The sphingolipid metabolic pathway and the glycerophospholipid metabolic pathway was considered a crucial signaling pathway by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. In addition, phosphatidylcholine and sphingomyelin were considered as hub metabolites. Twelve enzymes and 38 genes were considered as potential targets in the network analysis. Based on the combined analysis above, HJ was shown to modulate 2 key upstream targets, including PLA2G2A and PLA2G4A. Molecular docking showed that active compounds of HJ had high binding affinity with these key targets. In conclusion, the flavonoid components of HJ can inhibit PLA2 and regulate glycerophospholipid and sphingolipid metabolism pathway to delay the pathological process of ALI, which may be a potential mechanism of HJ against ALI.

Reducing COVID-19 diagnostic errors with dNTPαSe supplementation.

Timely and accurate diagnosis of COVID-19 is critical for controlling the pandemic. As the standard method to diagnose SARS-CoV-2, the real-time reverse transcription polymerase chain reaction (RT-qPCR) has good convenience. However, RT-qPCR still has a relatively high false-negative rate, particularly in the case of detecting low viral loads. In this study, using selenium-modified nucleoside triphosphates (dNTPαSe) in the RT-PCR reactions, we successfully increased the detection sensitivity and reduced the false-negative rate in COVID-19 diagnosis. By detecting positive controls, pseudovirus, and clinical samples with the commercial kits, we found that the dNTPαSe supplementation to these kits could generally offer smaller Ct values, permit the viral detection even in single-digit copies, and increase the detection specificity, sensitivity, and accuracy, thereby reducing the false-negative rate. Our experimental results demonstrated that dNTPαSe supplementation can make the commercial kits more specific, sensitive, and accurate, and this method is a convenient and efficient strategy for the disease detection and diagnosis.

Investigation of pharmacodynamic material basis of Anemarrhenae Rhizoma and its processed products based on plant metabolomics and molecular docking technology.

RATIONALE: Anemarrhenae Rhizoma (AR) has been an often used traditional Chinese medicine (TCM) for a long time. Its salt-processed form is one of the most common application forms. Modern pharmacological research has shown that the salt-processed product has various significantly enhanced pharmacological activities. However, the pharmacodynamic material basis of this change is not yet known. The aim of this study was to develop a strategy to screen pharmacodynamic substances in AR and salt-processed AR (SAR). METHODS: An integrated strategy combining plant metabolomics with molecular docking technology was established to screen pharmacodynamic substances. The plant metabolomics analysis was performed to select the chemical markers between AR and SAR. Then, molecular docking technology was applied to explore the relationship between chemical markers and diabetes targets (α-glucosidase). Finally, potential quality control markers were screened. RESULTS: There were significant differences in the quantification of nine steroidal saponins between AR and SAR. The results of plant metabolomics analysis showed a quite clear discrimination including 29 chemical markers between AR and SAR. Taking the hypoglycemic activity into consideration, 16 steroidal saponins were selected as potential quality markers. CONCLUSIONS: The developed method not only supplied an optional solution to search for pharmacophores in AR and SAR, but also provided a foundation for the study of the differential components and pharmacodynamics in various processed products of TCMs.

Characterization of Fritillariae cirrhosae bulbus from multiple sources by potential Q-marker based on metabolomics and network pharmacology.

RATIONALE: Fritillaria cirrhosae bulbus (BFC), a typical traditional Chinese medicine with multiple botanical sources, has been used for relieving cough and reducing sputum. Studies have shown that there were obvious differences in the chemical compositions and clinical efficacy of different sources of BFC. How to characterise BFC from botanical sources accurately and quickly is vital for drug quality evaluation and clinical applications. METHODS: In the present study, an integrated strategy of plant metabolomics combined with the target network pharmacology was developed to characterise BFC. Plant metabolomics analysis was performed to screen out the chemical markers of six species of BFC. Then, target network pharmacology was applied to explore the relationship between chemical markers and related diseases. Finally, potential Q-markers for species characterization were selected by combined analysis of plant metabolomics and the target network pharmacology. RESULTS: A total of 67 Fritillaria alkaloid compounds were identified. Six species showed clear characterization by multivariate statistical analysis, resulting in 12 chemical markers. Meanwhile, a total of nine components related to asthma were screened out based on the target network pharmacology. Taking content difference and pharmacological activity into consideration, nine constituents were selected as potential Q-markers. CONCLUSION: The method developed provided not only a standard protocol for characterising different species of BFC directly, but also an effective approach for multisource medicines discrimination.

A holistic comparison of flavor signature and chemical profile in different harvesting periods of Chrysanthemum morifolium Ramat. based on metabolomics combined with bioinformatics and molecular docking strategy.

Taiju and Duoju are products of Hangbaiju (HJ) obtained during different collection periods, and they have been commonly used as ingredients in tea beverages and dietary traditional Chinese medicine. This study reports an integrated strategy based on metabolomics, bioinformatics and molecular docking to further explore the effect of the harvesting period on the metabolic profile and clinical efficacy of HJ. Firstly, gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) were employed for non-targeted metabolomics profiling of essential oils and flavonoids. A sequential window acquisition of all theoretical fragment-ion spectra information-dependent acquisition (SWATH-IDA) bi-directionally verified (SIBDV) method was developed that integrates the advantages of both SWATH and IDA in characterizing flavonoids. Chemometric methods were then used to screen potential chemical markers. Furthermore, HJ is effective in hepatoprotective functions. Therefore, hepatocellular-carcinoma-related differentially expressed genes were obtained using bioinformatics, and the corresponding proteins were molecularly docked with diagnostic chemical markers. In total, 78 volatile oils and 63 flavonoids were tentatively identified. The results allowed the selection of 11 metabolites (5 volatile oils and 6 flavonoids), which are nominated as novel markers for material authentication of Taiju and Duoju. Additionally, two proteins associated with hepatoma were screened using bioinformatics. All six flavonoid markers with binding energies of <-5 kcal mol-1 were considered to be anti-hepatoma biomarkers. Noticeably, in Taiju, the content of hydroxygenkwanin showed a downward trend, but the content of the other five flavonoids and the five flavored volatile difference compounds had an upward trend. This bestows a unique flavor profile on Taiju, leading to differences in sensory aroma and clinical efficacy in Taiju and Duoju. In conclusion, the transformation of secondary metabolites was the dominant trend during HJ growth. These findings lay the foundation for food development and distinguishing clinical applications.

Qu-Du-San-Jie decoction induces growth inhibition and vascular normalization in NF2-associated vestibular schwannoma.

Background: Neurofibromatosis type 2 (NF2) is a rare genetic syndrome that predisposes individuals to develop bilateral vestibular schwannomas (VSs) causing a high risk of life-threatening neurological complications. Traditional treatment options for NF2-associated VS usually cause neurological damage, and to date, there are no FDA-approved pharmacotherapies for NF2. The aim of this study was to evaluate the antitumor efficacy of Qu-Du-San-Jie (QDSJ) decoction, a traditional Chinese medicine formula, on NF2-associated VS and to investigate the potential underlying mechanisms. Methods: Ultra high-performance liquid chromatography-mass spectroscopy (UHPLC-MS) analysis was performed to identify the components of QDSJ and their targets. To determine the relationships between the putative targets of QDSJ and the differential genes of NF2-associated VS, the drug-disease crossover genes were screened using the UHPLC-MS data combined with our previous gene expression profiling data. The differentially expressed genes were imported into the STRING database to generate a PPI network. Differentially expressed gene targets and pathways were identified using GO and KEGG pathway enrichment analyses. The in vitro and in vivo drug efficacy of QDSJ decoction was tested using a patient-derived schwannoma cell line and a patient-derived xenograft mouse model, respectively. H&E staining, immunochemistry, and immunofluorescence staining were used to evaluate the cell proliferation and tumor vessels. Results: A total of 133 compounds were identified in QDSJ decoction using UHPLC-MS analysis. Network pharmacology showed that the regulation of necroptosis, apoptosis, cell cycle, angiogenesis, adherens junction, and neuroactive ligand-receptor interaction could be associated with the efficacy of QDSJ in treating NF2-associated VS. Treatment with QDSJ induced necrotic cell death and apoptosis of schwannoma cells in vitro and suppressed the tumor growth in vivo. Histopathological analysis revealed areas of cell necrosis and enlarged tumor blood vessels in the QDSJ-treated tumors. The numbers of cells positive for Cyclin D1 and Ki-67 were significantly reduced in QDSJ-treated tumors compared to control tumors. Immunofluorescence staining of CD31 and αSMA showed a decreased number and density of tumor vessels and normalized vessel structure in QDSJ-treated tumors. Conclusion: Our study demonstrates that QDSJ decoction shows significant antitumor activity against NF2-associated schwannoma and is a possible candidate for future clinical trials.

An integrated strategy to reveal the potential anti-asthma mechanism of peimine by metabolite profiling, network pharmacology, and molecular docking.

Peimine, one of the major quality markers in Fritillaria Cirrhosae Bulbus, was expected to become a new anti-asthma drug. However, its metabolic profiles and anti-asthma mechanism have not been clarified previously. In this study, a method was developed for the detection of peimine metabolites in vitro by ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry. The potential anti-asthma mechanism was predicted by an integrated analysis of network pharmacology and molecular docking. A total of 19 metabolites were identified with the aid of software and molecular networking. The metabolic profiles of peimine elucidated that the metabolism was a multi-pathway process with characteristics of species difference. The network pharmacology results showed that peimine and its metabolites could regulate multiple asthma-related targets. The above targets were involved in various regulatory pathways linked to asthma. Moreover, the results of molecular docking showed that both peimine and its metabolites had a certain affinity with the ß2 adrenergic receptor. The results provided not only important references to understand the metabolism and pharmacodynamic changes of peimine in vitro, but also supporting data for further pharmacological evaluation. It also provided a new perspective for clarifying the functional changes of traditional Chinese medicine in vitro.

Multidisciplinary treatment of pancreatic cancer: a case report.

Background: Carcinoma of the head of pancreas has a high malignant degree and the 5-year survival rate at 5%. For unresectable pancreatic cancer, the 5-year survival rate is even lower. The clinical diagnosis of pancreatic cancer is difficult, and surgical indications are difficult to grasp. Moreover, perioperative and postoperative management is complex, and patients with different conditions need more attention to implement a comprehensive diagnosis and treatment strategy. In the diagnosis and treatment of pancreatic cancer and even other cancers, multi-disciplinary diagnosis and treatment can provide reasonable, personalized and more effective plans for patients so that cancer patients can receive better treatment and improve their quality of life. The multi-disciplinary diagnosis and treatment model can respond to the complex needs to individual patients. Case Description: This model is designed according to each patient's comprehensive situation, including their clinical symptoms, biochemical indicators, body mass index, and psychological status, and the tumor position, pathological tissue typing, and invasion scope. Complications of tumors can be reduced if treatment is controlled and if radical treatments are used within a broader comprehensive care model, thereby improving the quality of life of patients to prolong their survival. In our case report, the overall survival is much longer than un-resectable pancreatic cancer (median overall survival 6-9 months. The female patient was 53 years old. Her chief complaints were yellow skin, yellow urine, and absorption emaciation for 1 month. The preliminary diagnosis was postoperative pancreatic cancer. CT reexamination suggested portal vein liver metastasis. Repeated gastrointestinal bleeding occurs over the course of the disease. Postoperative review suggested recurrence, and she was admitted to First Affiliated Hospital, Army Medical University. During the treatment, the disease progressed to gastrointestinal bleeding, ascites, and jaundice. Conclusions: After multidisciplinary treatment (MDT) discussion, targeted treatment strategies were developed to improve the symptoms and improve the quality of life of the patients.

Robustness and accuracy evaluation of moisture prediction model for black tea withering process using hyperspectral imaging.

The rapid and non-destructive detection of moisture in withering leaves is an unsolved problem because the leaves are stacked together and have random orientation. To address this issue, this study aimed to establish more robust and accurate models. The performance of front side, back side and multi-region models were compared, and the front side model showed the worst transferability. Therefore, five effective wavelength (EW) selection algorithms were combined with a successive projection algorithm (SPA) to select EWs. It was found that the shuffled frog leaping algorithm (SFLA) combined with SPA was the best method for the front side model for moisture analyses. Based on the selected EWs, the extreme learning machine (ELM) became the model with the best self-verification result. Subsequently, moisture distribution maps of withering leaves were successfully generated. Considering the processing demand of withering leaves, local region models developed based on partial least squares and the SFLA-SPA method were applied to predict the moisture of withering leaves in the local and stacked region. The results showed that the RPD, Rcv and Rp values were above 1.6, 0.870 and 0.897, respectively. These results provide a useful reference for the non-destructive detection of moisture in withering leaves.

Effect of Electroacupuncture Treatment at Dazhui (GV14) and Mingmen (GV4) Modulates the PI3K/AKT/mTOR Signaling Pathway in Rats after Spinal Cord Injury.

Electroacupuncture (EA) is widely recognized as clinical treatment of spinal cord injury (SCI). The purpose of this study is to elucidate whether and how the PI3K/AKT/mTOR signaling pathway plays any role in EA treating SCI. Rats were randomly divided into four equal groups: Control Group, Sham-operation Group, Model Group, and EA Group, then further randomly divided into the following subgroups: 1-day (n = 12), 1-day rapamycin (n = 6), 14-day (n = 18), and 28-day (n = 18). A rat model of SCI was established by a modified Allen's weight-drop method. In the EA Group, rats were stimulated on Dazhui (GV14) and Mingmen (GV4) for 20 min by sterilized stainless steel needles. In the EA Group, the Basso, Beattie, and Bresnahan locomotor rating scale showed obvious improved locomotor function, and hematoxylin-eosin staining and magnetic resonance imaging showed that the histological morphology change of injured spinal cord tissue was obviously alleviated. Also, blocking spinal mTOR by injection of rapamycin showed that mTOR existed in the injured spinal cord, and EA could significantly activate mTOR in SCI rats. And immunohistochemistry and western blot analysis on the PI3K/AKT/mTOR signaling pathway showed that levels of PI3K, AKT, mTOR, and p70S6K in the injured spinal cord tissue were greatly increased in the EA Group, while the levels of PTEN and caspase 3 were decreased. The present study suggests that EA could affect cell growth, apoptosis, and autophagy through the PI3K/AKT/mTOR signaling pathway.

Effects of Paeonia lactiflora Extract on Estrogen Receptor ß, TPH2, and SERT in Rats with PMS Anxiety.

This study is to investigate the effect of Paeonia lactiflora extract on PMS anxiety and on expression of estrogen receptor ß (ERß), tryptophan hydroxylase-2 (TPH2), and serotonin transporter (SERT) in the premenstrual syndrome (PMS) anxiety model rats. The vaginal smear and open field test were used to screen rats in nonreception phase of estrus cycle with similar macroscopic behaviors and regular estrus cycle. PMS anxiety model rats were prepared by electrical stimulation. RT-PCR and immunofluorescence were used to measure the expression of ERß, TPH2, and SERT. Compared with normal rats, the total distance in the open field test of the model rats was significantly increased (P < 0.05). The model rats showed nervous alertness, irritability, and sensitivity to external stimuli. After treatment with the Paeonia lactiflora extract, the total distance of rats was significantly reduced (P < 0.05). In reception stage, there was no significant difference in the mRNA and protein expression of ERß, TPH2, and SERT. In nonreception stage, the expression of ERß and TPH2 in the model group was significantly decreased (P < 0.05) as compared with the control group, but not SERT. Abnormal changes of the above indicators were reversed after the administration of the Paeonia lactiflora extract. In conclusion, Paeonia lactiflora extract can increase the expression of ERß and TPH2 and decrease SERT in PMS model rats, which may be one of the mechanisms underlying the effect of Paeonia lactiflora extract on PMS.

Advances in Research of Drugs for Treating Respiratory Syncytial Virus / 中国实验方剂学杂志

Respiratory syncytial virus (RSV) is an RNA virus, which belongs to the paramyxoviridae family, and is transmitted by air droplets and close contact and the main pathogen causing acute lower respiratory tract infection in infants, the elderly and immunocompromised individuals.Although there have been studies on the prevention and treatment of RSV drugs and RSV infection in patients, many medical demands have not been met.And there is no specific antiviral therapy.The only two drugs approved to be applied in RSV prevention and treatment are perizol and ribavirin.However, the former must be used prophylactically, and primarily in high-risk children, while the latter is less effective, and some children even suffer from airway spasm.Therefore, it is urgent to propose new methods for prevention and treatment of RSV.In recent years, traditional Chinese medicine(TCM) has shown a good anti-RSV effect, with a fewer side effects, less resistance to drugs and broad-spectrum antiviral advantage.There are also newly developed biological and chemical anti-RSV drugs.Some new drugs have shown a good efficacy, with an extended half-life and reduced costs, such as fusion inhibitors, monoclonal antibodies.This paper reviews the research progress of anti-RSV drugs in the aspects of TCM, biological drug and chemical drugs, laying a foundation for the development of new anti-RSV drugs and the formulation of new therapeutic strategies.

[Correlation Study on Pathological Characteristics of Target Organs and Excess Evil Syndrome in IgA Nephropathy].

OBJECTIVE: To explore the correlation between pathological characteristics of target organs and excess evil syndrome in IgA nephropathy. METHODS: Data were collected in multicenter cooperation. Totally 266 IgA nephropathy patients were typed into exogenous wind-heat affection syndrome (49 cases), lower energizer damp-heat syndrome (100 cases), damp-phlegm syndrome (43 cases), and blood stasis syndrome (74 cases). Meanwhile, percutaneous renal biopsy was performed in all patients for Hass classification, Oxford classification, Katafuchi integral, and Jiang's classification methods. The correlation between excess evil syndrome and pathological index was analyzed. RESULTS: Four syndrome types were correlated with their Hass levels (r = 0. 341, P <0. 01). Affection of exogenous wind-heat syndrome was correlated with segmental proliferation of endothelial cells and damaged active lesions of segmental capillary loops. Lower-energizer damp-heat syndrome was associated with Hass III level, destroying active lesions of capillary loops, segmental proliferation of endothelial cells, glomerular segmental lesions, focal interstitial infiltration of inflammatory cells, focal interstitial fibrosis and tubular atrophy. Blood stasis syndrome was associated with Hass IV level, glomerular sclerosis, segmental glomerulosclerosis (S)/adhesion, mesangial hypercellularity (M), angiohyalinosis, multi-foci interstitial infiltration of inflammatory cells, multi-foci interstitial fibrosis and tubular atrophy. Phlegm-damp syndrome had higher proportions of Hass I and III levels, but with no association with other pathological parameters. CONCLUSIONS: Excess evil syndrome was associated with partial pathological characteristics of IgA nephropathy. It could reflect pathological damage degree of target organs, activities, chronic lesions, and prognosis of IgA nephropathy to certain extent. Correlated pathological characteristics and its evolution could indicate excess evil syndrome types and their evolution rules.

Correlation Study on Pathological Characteristics of Target Organs and Excess Evil Syndrome in IgA Nephropathy / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the correlation between pathological characteristics of target organs and excess evil syndrome in IgA nephropathy.</p><p><b>METHODS</b>Data were collected in multicenter cooperation. Totally 266 IgA nephropathy patients were typed into exogenous wind-heat affection syndrome (49 cases), lower energizer damp-heat syndrome (100 cases), damp-phlegm syndrome (43 cases), and blood stasis syndrome (74 cases). Meanwhile, percutaneous renal biopsy was performed in all patients for Hass classification, Oxford classification, Katafuchi integral, and Jiang's classification methods. The correlation between excess evil syndrome and pathological index was analyzed.</p><p><b>RESULTS</b>Four syndrome types were correlated with their Hass levels (r = 0. 341, P <0. 01). Affection of exogenous wind-heat syndrome was correlated with segmental proliferation of endothelial cells and damaged active lesions of segmental capillary loops. Lower-energizer damp-heat syndrome was associated with Hass III level, destroying active lesions of capillary loops, segmental proliferation of endothelial cells, glomerular segmental lesions, focal interstitial infiltration of inflammatory cells, focal interstitial fibrosis and tubular atrophy. Blood stasis syndrome was associated with Hass IV level, glomerular sclerosis, segmental glomerulosclerosis (S)/adhesion, mesangial hypercellularity (M), angiohyalinosis, multi-foci interstitial infiltration of inflammatory cells, multi-foci interstitial fibrosis and tubular atrophy. Phlegm-damp syndrome had higher proportions of Hass I and III levels, but with no association with other pathological parameters.</p><p><b>CONCLUSIONS</b>Excess evil syndrome was associated with partial pathological characteristics of IgA nephropathy. It could reflect pathological damage degree of target organs, activities, chronic lesions, and prognosis of IgA nephropathy to certain extent. Correlated pathological characteristics and its evolution could indicate excess evil syndrome types and their evolution rules.</p>

[Screening of 10 types of Chinese herbal compounds inhibiting Abeta and their possible related mechanism in vitro].

This study is to screen the Chinese herbal compounds which could inhibit the production of Abeta and investigate the underlying mechanism. Ten types of compounds which have potential value in the treatment of AD were selected as initial screening trial. The cell models which used could overexpress Abeta and beta-secretases or Abeta and gamma-secretases. Extracellular Abeta was determined by ELISA after the cell models treated with different concentrations of compounds (0.5-100 micromol x L(-1)), separately. Then the compounds were selected which could inhibit extracellular Abeta and their best concentration ranges were decided, too. Furthermore, the cell viability and apoptosis rate, the level of intracellular Abeta, beta and gamma-secretases were determined after the cell models treated with different concentrations of selected compounds. The results showed that 4 of the 10 compounds could reduce the level of extracellular Abeta; they were cryptotanshinone, astragalosides, gastrodin and paeoniflorin, and their best concentration ranges were 0.5-5.0, 0.5-5.0, 5.0-50, 1.0-25 micromol x L(-1), respectively. Further study indicated that the 4 selected compounds were nontoxic to the cellular models and lowering intracellular Abeta were more effective compared with extracellular; of which astragalosides and gastrodin showed dose-dependent inhibition to the activities of beta and gamma-secretases, with the maximum inhibiting rates of 78.2% and 80.3%, respectively. In conclusion, cryptotanshinone, astragalosides, gastrodin and paeoniflorin could inhibit the expression and secretion of Abeta, and the underlying inhibiting mechanism of astragalosides and gastrodin were related with the reduction of the beta and gamma-secretase activities, respectively.

Screening of 10 types of Chinese herbal compounds inhibiting Abeta and their possible related mechanism in vitro / 药学学报

This study is to screen the Chinese herbal compounds which could inhibit the production of Abeta and investigate the underlying mechanism. Ten types of compounds which have potential value in the treatment of AD were selected as initial screening trial. The cell models which used could overexpress Abeta and beta-secretases or Abeta and gamma-secretases. Extracellular Abeta was determined by ELISA after the cell models treated with different concentrations of compounds (0.5-100 micromol x L(-1)), separately. Then the compounds were selected which could inhibit extracellular Abeta and their best concentration ranges were decided, too. Furthermore, the cell viability and apoptosis rate, the level of intracellular Abeta, beta and gamma-secretases were determined after the cell models treated with different concentrations of selected compounds. The results showed that 4 of the 10 compounds could reduce the level of extracellular Abeta; they were cryptotanshinone, astragalosides, gastrodin and paeoniflorin, and their best concentration ranges were 0.5-5.0, 0.5-5.0, 5.0-50, 1.0-25 micromol x L(-1), respectively. Further study indicated that the 4 selected compounds were nontoxic to the cellular models and lowering intracellular Abeta were more effective compared with extracellular; of which astragalosides and gastrodin showed dose-dependent inhibition to the activities of beta and gamma-secretases, with the maximum inhibiting rates of 78.2% and 80.3%, respectively. In conclusion, cryptotanshinone, astragalosides, gastrodin and paeoniflorin could inhibit the expression and secretion of Abeta, and the underlying inhibiting mechanism of astragalosides and gastrodin were related with the reduction of the beta and gamma-secretase activities, respectively.

Application of an enzyme immunoassay for the quantitative determination of azo dye (Orange II) in food products.

This paper reports the preparation of polyclonal antibodies against a synthetic azo dye, Orange II, and the development of an indirect ELISA to detect Orange II in foods. The sulfonic group of Orange II was modified and linked with carrier protein to synthesise an artificial antigen. Based on the checkerboard titration, the method showed excellent sensitivity (IC50 = 0.61 ng g⁻¹) to Orange II in the linear range of 0.05-10 ng g⁻¹. The antibody had little cross-reactivity with Chromotrope FB, Gardenia Yellow, Ponceau 4R, Sunset Yellow and Sudan dyes. The ELISA had limits of detection (LOD) of 0.22, 0.97 and 0.74 ng g⁻¹ in chilli powder, chilli oil and braised pork, respectively. The limits of quantification (LOQ) of the assay were 0.91 ng g⁻¹ in chilli powder, 1.48 ng g⁻¹ in chilli oil and 1.10 ng g⁻¹ in braised pork. For food products fortified with 1-10 ng g⁻¹ Orange II, the inter- and intra-assay variations were all less than 24.0% and 18.0%, respectively. Therefore, the proposed test could be used as a rapid screening method for Orange II detection in food samples.

Determination of folic acid in milk, milk powder and energy drink by an indirect immunoassay.

BACKGROUND: Folic acid (FA) is essential for healthy people (reference daily intake 400 µg day⁻¹) and pregnant women (600 µg day⁻¹). Insufficient intake of FA will increase the risk of neural tube defects in newborns. In this study an indirect enzyme-linked immunosorbent assay was developed for rapid and convenient detection of FA in vitamin-fortified foods. RESULTS: A carbodiimide-modified active ester method was used to synthesise the immunogen (FA-bovine serum albumin (BSA) conjugate) to raise polyclonal antibodies for FA. The coupling ratio of FA with BSA was determined to be 14:1 (molar ratio). The detection limit of the immunoassay was 3.0 ng mL⁻¹ in buffer, 3.52 ng mL⁻¹ in energy drink, 11.91 ng mL⁻¹ in milk and 16.50 ng mL⁻¹ in milk powder. Intra- and inter-assay variability ranged from 6.6 to 15.1%. Analytical recoveries of FA-spiked samples were 88.3-108.9%. CONCLUSION: The immunoassay developed in this study can be used as a simple, rapid and accurate method for fast semi-quantitative and quantitative on-site analysis of FA in food products.