Rapid screening and evaluation of natural antioxidants from leaf, stem, and root of Artemisia argyi by online liquid microextraction combined with HPLC-based antioxidant assay system coupled with calibration quantitative analysis.

To reveal the utilization value of leaf, stem, and root of Artemisia argyi, a rapid online liquid microextraction combined with a high-performance liquid chromatography coupled with 2,2-nitrogen-di (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt antioxidant assay system was established for analysis of antioxidants in the leaf, stem, and root of A. argyi, and a calibration quantitative method of antioxidant activity with equivalent chlorogenic acid was proposed. Thirty-three positive peaks were identified; among them, 12 compounds were found that possess good antioxidant activity including eleven organic acids (components 2-4, 8, 11-14, 17, 19, and 21) and one flavonoids (component 22). The proposed calibration quantitative method avoided the influence of content of compound and compared the extent of radical scavenging capacity of five antioxidant compounds, which were ranked as follow: 3,5-dicaffeoylquinic acid > 3,4-dicaffeoylquinic acid ≈ 4,5-dicaffeoylquinic acid > 1,4-dicaffeoylquinic acid > chlorogenic acid. In conclusion, this study provided composition and biological potential for the future development of the leaf, stem, and root of A. argyi. It is believed that the online liquid microextraction combined with high-performance liquid chromatography based antioxidant assay system can be widely used for the rapid screening of natural antioxidant components in the different parts of natural products.

Identification and quantification of nine compounds in Fangwen Jiuwei decoction by liquid chromatography-mass spectrometry.

Fangwen Jiuwei Decoction is a traditional Chinese medicine preparation for the treatment of pneumonia developed by Shenzhen Bao'an Chinese Medicine Hospital, which shows remarkable clinical responses. Qualitative and quantitative analyses of the main active compounds are crucial for the quality control of traditional Chinese medicine prescription in clinical application. In this study, we identified nine active compounds essential for the pharmacological effects of Fangwen Jiuwei Decoction based on the analysis of the Network Pharmacology and relevant literature. Moreover, these compounds can interact with several crucial drug targets in pneumonia based on molecular docking. We applied high-performance liquid chromatography-tandem mass spectrometry method was established these nine active ingredients' qualitative and quantitative detections. The possible cleavage pathways of nine active components were determined based on secondary ions mass spectrometry. The results of high-performance liquid chromatography-tandem mass spectrometry were further validated, which show a satisfactory correlation coefficient (r > 0.99), recovery rate (≥93.31%), repeatability rate (≤5.62%), stability (≤7.95%), intra-day precision (≤6.68%), and inter-day precision (≤9.78%). The limit of detection was as low as 0.01 ng/ml. In this study, we established a high-performance liquid chromatography-tandem mass spectrometry method to qualitatively and quantitatively analyze the chemical components in the Fangwen Jiuwei Decoction extract.

Determination of six core components from Mahuang Xuanfei Zhike syrup in rat plasma and tissues by UPLC-MS/MS: Application to a pharmacokinetics and tissue distribution study.

Mahuang Xuanfei Zhike (MXZ) syrup, a Chinese patent medicine, has been widely used in the clinical treatment of cough. However, there is no reported method for the quantitative analysis of the effective components of MXZ syrup in biological samples. In this study, the effective components of MXZ syrup were screened by network pharmacology and molecular docking technology. A sensitive and rapid ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established to test the active components of MXZ syrup in rat plasma and tissue homogenates, including ephedrine, amygdalin, chlorogenic acid, harpagoside, forsythin and forsythoside A. Chromatographic separation was performed on a Waters Acquity UPLC HSS T3 column (2.1 × 50 mm, 1.8 µm) and the mass analysis was conducted using a Waters Xevo TQ mass spectrometer using multiple reaction positive and negative ion simultaneous monitoring mode. The results showed that the linearity ranged from 0.3 to 409.4 ng/ml. The extraction recoveries were all <8.33%, and the matrix effects were all <8.45, which met the requirements. The pharmacokinetic and tissue distribution results indicated that the main active components of MXZ syrup were absorbed quickly and eliminated slowly in vivo, and there may be a reabsorption process.

[Research progress in chemical constituents in plants of Polygonatum and their pharmacological effects].

Polygonatum plants are perennial plants of Liliaceae. There are about 60 species reported at home and abroad,32 species in China,mainly distributed in the north temperate zone. Their main chemical components are steroidal saponins,polysaccharides,flavonoids,alkaloids,etc. They have anti-aging,anti-tumor,immunomodulation,antibacterial,antiviral,hypoglycemic and blood lipid effects. With the development of health industry,Polygonati Rhizome used as medicine and food has attracted great attention in recent years,and has become a research hotspot. However,the material basis of its efficacy is unclear and the product quality is uneven,which seriously limited the rapid upgrading of the industry. This review summarizes Polygonatum plants system classification,the chemical composition and pharmacological activity to provide theoretical basis for the development and utilization of Polygonatum plants.

[Study on HPLC Fingerprint of Solanum nigrum Fruit].

Objective: To establish the HPLC fingerprint method of Solanum nigrum fruit for its identification and quality evaluation. Methods: The analysis was carried out by Agilent ZORBAX SB-C18column( 150 mm × 4. 6 mm,5 µm),and eluted with mobile phase containing of acetonitrile-0. 3% phosphoric acid in a gradient mode. The temperature of column was 25 ℃,the flow rate was 1. 0m L / min, the detection wavelength was set at 205 nm, the injection volume was 10 µL. The similarity evaluation system for chromatographic fingerprint of TCM( version 2004A) was used to calculate the similarity degree of the HPLC fingerprint of Solanum nigrum fruit. . Results: The HPLC fingerprint of ten batches Solanum nigrum fruit was established with twelve common peaks. Two characteristic peaks included solasonine and solamargine were confirmed. Conclusion: The results indicate that establishing HPLC fingerprint of Solanum nigrum fruit can provide more comprehensive reference for identification and quality evaluation of Solanum nigrum fruit

[Study on Triterpenoids from Helicteres angustifolia by High-Speed Countercurrent Chromatography].

Objective: To develop a method for separation and purification of triterpenoids from Helicteres angustifolia by highspeed countercurrent chromatography( HSCCC). Methods: The ethyl acetate extract of Helicteres angustifolia were separated by two-step HSCCC. The structures of the target compounds were identified by ESI-MS and1H-NMR and characterized by FTIR. Results: Six compounds were isolated and prepared from ethyl acetate extract of Helicteres angustifolia,and identified as methyl helicterilate( 1),methyl helicterate( 2),helicterilic acid( 3),helicteric acid( 4),betulic acid( 5),oleanolic acid( 6). The purity of the chemical constituents were determined by HPLC,and the mass fraction was more than 95. 0%,respectively. Conclusion: This method is suitable for the separation and preparation of triterpenoids from Helicteres angustifolia,which is rapid,high purity and high yield.

[Study on Chemical Constituents of Petroleum Ether Fraction from Rubus alceaefolius].

OBJECTIVE: To investigate the chemical constituents of Rubus alceaefolius. METHODS: Nine compounds were isolated and purified from the petroleum ether extract of 95% alcohol extract of Rubus alceaefolius by repeated column chromatography on silica, Sephadex LH-20 and structurally identified by spectral analysis. RESULTS: The compounds were identified as chrysophanol(1), physcion (2), ß-sitosterol(3), 3-oxotirucalla-7, 24-dien-21-oic acid(4), myricadiol(5), 19-α-hydroxy-3-acetyl-ursolic acid(6), N-benzoylphenylalaninyl-N-benzoylphenylalaninate(7), aurantiamide acetate(8) and euscaphic acid(9). CONCLUSION: Compounds land 4~8 are isolated from this plant for the first time, and compounds 4 - 8 are found in plants of Rubus genus for the first time.

[HPLC Fingerprints of Clerodendrum lindleyi].

OBJECTIVE: To establish the HPLC fingerprint of Clerodendrum lindleyi in order to provide the basis for its quality standard. METHODS: The chromatographic fingerprint was obtained with Angilent Zorbax C18 (250 mm x 4.6 mm, 5 µm) column and gradiently eluted with acetonitrile-0.1% phosphoric acid solution. The column temperature was maintained at 35 degrees C. The flow rate was 1.0 mL/min and the detection wavelength was 327 nm. RESULTS: HPLC fingerprint of Clerodendrum lindleyi was established and 21 common peaks from 11 batches of samples were found. CONCLUSION: The method has good precision, stability and repeatability, which can provide reliable basis for quality evaluation of Clerodendrum lindleyi.

[Chemical constituents from roots of Rubus parvifolius].

OBJECTIVE: To investigate the chemical constituents from the roots of Rubus parvifolius. METHODS: The chemical constituents were isolated by silica gel column chromatography, Sephadex LH-20, as well as preparative high performance liquid chromatography. Their structures were identified on the basis of physicochemical properties and NMR analysis. RESULTS: Six comounds were isolated and identified as 3-O-Acetyl-11α, 12α-epoxy-oleanan-28,ß-olide( I ) ,3-O-Acetyl-pomolic acid( II ), Ursolic acid( Ill),Ursolic acid acetate (1V ), Euscaphic acid ( V) and ß-Sitosterol ( VI). CONCLUSION: Compounds I , II and IV are isolated from Rubus parvifolius for the first time.

Discovery of mitochondrion-targeting copper(II)-plumbagin and -bipyridine complexes as chemodynamic therapy agents with enhanced antitumor activity.

Four copper(II)-plumbagin and -bipyridine complexes (Cu1-Cu4) were synthesized as chemodynamic therapy agents with enhanced antitumor activity. As lipophilic and positively charged compounds, Cu1-Cu4 were preferentially accumulated in mitochondria and activated the mitochondrial apoptosis pathway. Mechanistic studies showed that Cu1-Cu4 reacted with GSH to reduce Cu2+ ions to Cu+ ions, catalyzed the formation of toxic hydroxyl radicals (˙OH) from hydrogen peroxide (H2O2) through a Fenton-like reaction, induced mitochondrial dysfunction, and activated caspase-9/3, which eventually led to apoptosis. Cu1-Cu4 arrested HeLa cells in the S phase and eventually killed cancer cells. Cu2 showed a favorable pharmacokinetic profile in mice. Moreover, Cu2 effectively inhibited the growth of HeLa xenografts in nude mice and showed low toxicity in vivo.

[Optimization extraction conditions of triphyllin A in Pronephrium triphyllum].

OBJECTIVE: To optimize the extraction conditions for triphyllin A in Pronephrium triphyllum. METHODS: The content of triphyllin A in the Pronephrium triphyllum was determined by HPLC. Concentration and volume of the alcohol,time and times of the extraction were assayed by orthogonal test to detect their influences on the extraction rate of triphyllin A in the Pronephrium triphyllum. RESULTS: Alcohol volume and extraction times had significant influence on the process (P < 0.05) while alcohol concentration and extraction time had no effect. The optimal extraction conditions were as follows:50 fold 60% alcohol, extraction for 3 times and 50 min for each time. CONCLUSION: The extration rate of triphyllin A is higher,and the process can be used for the development and production of Pronephrium triphyllum.

[Species and ecological control of disease on cultivated Gentiana rigescens in Yunnan].

OBJECTIVE: To find out variety of the fungal diseases of cultivated Gentiana rigescens and provide important basis for prevention. METHODS: The diseases were diagnosed based on field investigate, symptoms observation, pathogen isolation, determination the size of morphological and verification following the Koch's Postulate procedures. RESULTS: Anthracnose (Colletotrichum gloeosporioides), grey mould (Botrytis cinerea), brown spot (Alternaria tenuis), rust (Aecidiumpers), circular spot (Pestalotiopsis), leaf blight (Stemphylium, Ascochyta, Pleospora) and nematodes (Heterodera spp., Meloidogyne spp.) were found on Gentiana rigescens. Anthracnose was the first main disease, the diseased plant rate was over 40% and disease severity was 4 - 5 degree and second disease was rust, incidence of rate was less 10% and other diseases rate was not enough 2%. CONCLUSION: All these diseases on Gentiana rigescens are reported for the first time and Gentiana rigescens is the new host plant of the diseases.

[Effects of environmental enrichment on the repair and proliferation of neurons in neonatal rats with hypoxic-ischemic brain damage].

OBJECTIVE: To study the effects of environmental enrichment on neuron proliferation, learning and memory ability and motor ability in neonatal rats with hypoxic-ischemic brain damage (HIBD). METHODS: One hundred and eight 7-day-old Sprague-Dawley rats were randomly divided into three groups: sham operation (CON group), HIBD and intervention group. HIBD model was prepared according to the classic Rice-Vannucci method. Environmental enrichment was administered for the rats in the intervention group after HIBD inducement. Behavioral tests (Water maze test, Suspension test and Slope test) were performed and the number of neural cells in the left hippocampus was examined 7, 14 and 28 days after intervention. RESULTS: The pyramid cells in the hippocampus CA1 area in the HIBD group were significantly less than in the CON group at 7, 14 and 28 days (P<0.05). The number of pyramid cells in the hippocampus CA1 area in the intervention group was significantly higher than in the HIBD group (P<0.01) at 7, 14 and 28 days. The hidden platform escape latency period (EL) in the Water maze test was significantly more prolonged and the cross-platform number within 2 minutes was significantly less in the HIBD and the intervention groups than in the CON group at all observed time points (P<0.01). The EL was significantly shorter and the cross-platform number within 2 minutes was significantly higher in the intervention group than in the HIBD group at all observed time points (P<0.01). The maintain time and score in the Suspension test were significantly lower and the time in the Slope test was significantly more prolonged in the HIBD and intervention groups than in the CON group at 7, 14 and 28 days (P<0.01). An increased maintain time and score and a decreased time in the Slope test were found in the intervention group compared with the HIBD group at 14 and 28 days (P<0.01). CONCLUSIONS: Environmental enrichment can improve motor function, learning and memory ability, and promote the repair and proliferation of neurons in neonatal rats with HIBD.

Fraud Detection of Herbal Medicines Based on Modern Analytical Technologies Combine with Chemometrics Approach: A Review.

Fraud in herbal medicines (HMs), commonplace throughout human history, is significantly related to medicinal effects with sometimes lethal consequences. Major HMs fraud events seem to occur with a certain regularity, such as substitution by counterfeits, adulteration by addition of inferior production-own materials, adulteration by chemical compounds, and adulteration by addition of foreign matter. The assessment of HMs fraud is in urgent demand to guarantee consumer protection against the four fraudulent activities. In this review, three analysis platforms (targeted, non-targeted, and the combination of non-targeted and targeted analysis) were introduced and summarized. Furthermore, the integration of analysis technology and chemometrics method (e.g., class-modeling, discrimination, and regression method) have also been discussed. Each integration shows different applicability depending on their advantages, drawbacks, and some factors, such as the explicit objective analysis or the nature of four types of HMs fraud. In an attempt to better solve four typical HMs fraud, appropriate analytical strategies are advised and illustrated with several typical studies. The article provides a general workflow of analysis methods that have been used for detection of HMs fraud. All analysis technologies and chemometrics methods applied can conduce to excellent reference value for further exploration of analysis methods in HMs fraud.

The Actual Status of Hospitals as COVID-19 Vaccination Clinics in China and Safety Monitoring of Inactivated Vaccine: A Cross-Sectional Study.

BACKGROUND: The outbreak has had a devastating impact, and efforts are underway to speed up vaccination. The study's objective was to describe the clinical characteristics of the coronavirus disease 2019 (COVID-19) vaccination clinic in the Second People's Hospital of Fujian Province, China. Meanwhile, we monitored all the vaccine recipients to evaluate adverse reactions. METHODS: A cross-sectional study was done at the COVID-19 Vaccination Clinic, the Second People's Hospital of Fujian Province, China. We systematically collected Clinical data from the COVID-19 vaccination clinic between March 11 and November 11, 2021, including the type of vaccine, number of doses, gender, age, educational level, occupational category, adverse reactions, etc. Investigators will contact vaccine recipients by means of phone call or WeChat message to record the negative responses. Last, this report covers data through 8 mo, so it will be better to Evaluate the Safety of 2 inactivated COVID-19 vaccines from China (BBIBP-CorV [Beijing Institute of Biological Products, Beijing, China] and CoronaVac [Sinovac Life Sciences, Beijing, China]). RESULTS: The results indicated that the Second People's Hospital of Fujian Province received a total of 64,602 COVID-19 vaccines from March 11 to November 11, 2021, including 34,331 (53.14%) first doses, 29,245 (45.27%) second doses, and 1026 (1.59%) third doses. This study found the highest proportion in other personnel (38.69% at the first dose, 38.75% at the second dose, and 2.44% at the third dose), who were mainly retirees. People with higher levels of education are more likely to be vaccinated against COVID-19 during the early stages of vaccine rollout. In terms of age stratification, the highest proportion was found among people aged 18-49 (BBIBP-CorV: first dose 61%, second dose 62.6%, and third dose 76.8%; CoronaVac: first dose 66.1%, double dose 63.6%, and third dose 75.5%), followed by those over 60. The common adverse reactions were mainly local and systemic, and there were some differences between the 2 inactivated vaccines (P < 0.05). CONCLUSIONS: This is the first study to analyze the actual status of hospitals as COVID-19 vaccination clinics in China. The hospital has focused on vaccinating citizens and the initial rollout of vaccines to ensure any safety issues are identified. More citizens are willing to vaccinate in hospitals because of the uncertain safety of the available vaccines and adverse reactions. The good news is that vaccine-related severe adverse events have not been found in the hospital vaccination clinic. The Safety of BBIBP-CorV and CoronaVac is relatively high.

Effects of Different Pre-drying and Drying Methods on Volatile Compounds in the Pericarp and Kernel of Amomum tsao-ko.

The effects of twelve different pre-drying and drying methods on the chemical composition in the pericarp and kernel of Amomum tsao-ko were studied. The volatile components were isolated from the samples by simultaneous distillation and extraction and analyzed by gas chromatography-mass spectrometry (GC-MS). Sixty and thirty-eight compounds were identified from pericarp and kernel, respectively, and the main constituents were oxygenated monoterpenes. These compounds were not only significantly affected by pre-drying and drying methods but also varied in content due to different tissue locations. The total volatile content of pericarp varied from 0.70 to 1.55%, with the highest obtained by microwave-dried samples (150 W) and the lowest in freeze-dried samples. The total volatile content of the kernel varied from 6.11 to 10.69%, with the highest content obtained during sun drying (SD) and the lowest content in samples treated with boiling water for 2 min. Oxygenated monoterpenes were the highest compounds in pericarp and kernel, which were also the most affected by drying methods. The highest content of oxygenated monoterpenes in the pericarp (0.77%) could be obtained by boiling water treatment for 5 min, and the highest content of oxygenated monoterpenes in the kernel (7.48%) could be obtained by SD. Additionally, the main components such as 1,8-cineole, 2-carene, (Z)-citral, nerolidol, (Z)-2-decenal, (E)-2-dodecenal, citral, (E)-2-octenal, 4-propylbenzaldehyde, and phthalan showed remarkable variations in pre-drying and drying methods.

[Study on the chemical constituents from the aerial parts of Medicago sativa and their hypolipidemic activity].

OBJECTIVE: To research the chemical constituents from the aerial parts of Medicago sativa. METHODS: Various column chromatographies were emoployed to isolate and purify the constituents. Their structures were elucidated by spectral analysis (IR, UV, MS, 1H-NMR, 13C-NMR) and chemical evidence. RESULTS: Two constituents were obtained and identified as soysaponin I (I), azukisaponin V (II). CONCLUSION: These two compounds are isolated from the arial parts of Mesicago sativa for the first time. Moreover,the compounds are found to have hypolipidemic activity.

Construction and identification of a yeast two-hybrid bait vector and its effect on the growth of yeast cells and the self-activating function of reporter genes for screening of HPV18 E6-interacting protein.

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

Identification of Chinese Herbal Compounds with Potential as JAK3 Inhibitors.

The Janus kinases (JAKs) consist of four similar tyrosine kinases and function as key hubs in the signaling pathways that are implicated in both innate and adaptive immunity. Among the four members, JAK3 is probably the more attractive target for treatment of inflammatory diseases because its inhibition demonstrates the greatest immunosuppression and most profound effect in the treatment of such disorders. Although many JAK3 inhibitors are already available, certain shortcomings have been identified, mostly acquired drug resistance or unwanted side effects. To discover and identify new promising lead candidates, in this study, the structure of JAK3 (3LXK) was obtained from the Protein Data Bank and used for simulation modeling and protein-ligand interaction analysis. The ~36,000 Chinese herbal compounds obtained from TCM Database@Taiwan were virtually screened by AutoDock Vina docking program and filtered with Lipinski's Rules and ADME/T virtual predictions. Because of high occurrence of fake hits during docking, we selected 12 phytochemicals which have demonstrated modulating JAKs expressions among the top 50 chemicals from docking results. To validate whether these compounds are able to directly mediate JAK3 kinase, we have investigated the inhibitory activity using enzymatic activity assays, western blot, and HEK 293 cell STAT5 transactivity assays. The molecular analysis included docking and molecular dynamics (MD) simulations in order to investigate structural conformations and to explore the key amino acids in the interaction between JAK3 kinase and its putative ligands. The results demonstrated that Cryptotanshinone, Icaritin, and Indirubin exhibited substantial inhibitory activity against JAK3 kinase in vitro. The results also provide binding models of the protein-ligand interaction, detailing the interacting amino acid residues at the active ATP-binding domains of JAK3 kinase. In conclusion, our work discovered 3 potential natural inhibitors of JAK3 kinase and could provide new possibilities and stimulate new insights for the treatment of JAK3-targeted diseases.

Helicteric Acid, Oleanic Acid, and Betulinic Acid, Three Triterpenes from Helicteres angustifolia L., Inhibit Proliferation and Induce Apoptosis in HT-29 Colorectal Cancer Cells via Suppressing NF-κB and STAT3 Signaling.

Colorectal cancer (CRC) is one of the most common malignancies and most frequent cause of cancer death worldwide. The activation of both NF-κB and STAT3 signaling and the crosstalk between them play an important role in colorectal tumor. Helicteres angustifolia L. is a type of commonly used Chinese medicinal herb and possesses a wide variety of biological activities. In the present study, we investigate the effects of three triterpenes from H. angustifolia (HT) such as helicteric acid (HA), oleanic acid (OA), and betulinic acid (BA), on inhibiting CRC progression. Our results showed that HT extracts could decrease proliferation and induce apoptosis in HT-29 colorectal cancer cells. Moreover, HT extracts could suppress LPS-triggered phosphorylation of IKK, IκB, and NF-κB, attenuate IL-6-induced phosphorylation of JAK2 and STAT3, and suppress the expression of c-Myc, cyclin-D1, and BCL-xL, the downstream gene targets of NF-κB and STAT3. Therefore, HT extracts showed potent therapeutic and antitumor effects on CRC via inhibiting NF-κB and STAT3 signaling.