[Assay of amino acids in leaves of Eucommia ulmoides under arbor forest mode and leaf-oriented cultivation mode by pre-column derivatization HPLC].

This study aims to develop the pre-column derivatization high performance liquid chromatography(HPLC) method for the determination of 16 kinds of amino acids in Eucommia ulmoides leaves, and compare the content of amino acids in the leaves harvested at different time and under leaf-oriented cultivation mode(LCM) and arbor forest mode(AFM). The HPLC conditions are as below: phenyl isothiocyanate(PITC) as pre-column derivatization agent, Agilent ZORBAX C_(18 )column(4.6 mm×250 mm, 5 µm), mobile phase A of acetonitrile-water(80∶20), mobile phase B of 0.1 mol·L~(-1) sodium acetate solution-acetonitrile(94∶6), gradient elution, flow rate of 1.0 mL·min~(-1), injection volume of 5 µL, column temperature of 40 ℃, and detection wavelength of 254 nm. The HPLC profile indicated well separation of 16 kinds of amino acids and the amino acid content in E. ulmoides leaves was up to 16.26%. In addition, the amino acid content in leaves of E. ulmoides under LCM was higher than under AFM. The amino acid content varied with the harvesting time. Through orthogonal partial least squares discriminant analysis, the amino acids of E. ulmoides under LCM and AFM were compared, which can distinguish the leaves under LCM from those under AFM. Principal component analysis was applied to comprehensively score the amino acids of E. ulmoides leaves. The results showed that the score of leaves under LCM was higher than that under AFM. Nutritional evaluation results indicated that the proteins in E. ulmoides leaves belonged to high-quality vegetable proteins. The established method for the determination of amino acid content is reliable. With the amino acid content as index, the leaf quality of E. ulmoides under LCM is better than that under AFM. This study lays a theoretical basis for the promotion of LCM for E. ulmoides and the development of medicinal and edible products from E. ulmoides leaves.

[Effects of different drying methods on quality of male flowers of Eucommia ulmoides based on color and chemical composition].

To study the effects of different drying methods on the quality of male flowers of Eucommia ulmoides(MFOEU), we treated fresh MFOEU samples with drying in the shade(DS), vacuum freeze drying(VFD), high-or low-temperature hot air drying(HTHAD, LTHAD), microwave drying(MD), and vacuum drying(VD), respectively. The color, total flavonoid content, total polysaccharide content, and main active components such as geniposide, geniposidic acid, rutin, chlorogenic acid, galuteolin, pinoresinol diglucoside, and aucubin in MFOEU were taken as the evaluation indicators. The quality of MFOEU was comprehensively evaluated by entropy weight method combined with color index method, partial least squares discriminant analysis and content clustering heat map. The experimental results showed that VFD and DS basically kept the original color of MFOEU. The MFOEU treated with MD had higher content of total polysaccharides, phenylpropanoids, lignans, and iridoids. The MFOEU treated with LTHAD had higher content of total flavonoids and that treated with VD had lower content of active components. According to the results of comprehensive evaluation, the quality of MFOEU dried with different methods followed the order of MD>HTHAD>VFD>LTHAD>DS>VD. Considering the color of MFOEU, the suitable drying methods were DS and VFD. Considering the color, active components, and economic benefits of MFOEU, MD was the suitable drying method. The results of this study are of a reference value for the determination of suitable methods for MFOEU processing in the producing areas.

[Spleen deficiency and phlegm dampness syndrome model in rats treated by Citri Reticulatae Pericarpium based on metabolomics].

The present study analyzed the effect of Citri Reticulatae Pericarpium on endogenous metabolites in spleen deficiency and phlegm dampness syndrome by metabolomics, and explored the underlying mechanism of Citri Reticulatae Pericarpium in the treatment of spleen deficiency and phlegm dampness syndrome.The model of spleen deficiency and phlegm dampness syndrome was induced in rats by the multi-factor modeling method.The intervention effects of Citri Reticulatae Pericarpium on rats with spleen deficiency and phlegm dampness syndrome were preliminarily evaluated by observing the pathological changes of rat liver tissues and measuring the plasma content of pathological and biochemical indexes such as triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C).Immunohistochemistry was used to detect the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland, and the effect of Citri Reticulatae Pericarpium on aquaporin expression in rats with spleen deficiency and phlegm dampness syndrome was evaluated.Furthermore, UHPLC-ESI-MS/MS was used to analyze the metabolic profiles of rat plasma samples.Multiple methods, such as principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used for pattern recognition.Differential metabolites were screened out by t-test and variable importance in projection(VIP), followed by pathway analysis based on MetaboAnalyst 5.0.As revealed by experimental results, Citri Reticulatae Pericarpium could improve the pathological changes of liver tissues, increase the levels of HDL-C in the plasma, reduce the levels of TC, TG, and LDL-C, and enhance the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland of rats with spleen deficiency and phlegm dampness syndrome.In addition, 87 differential metabolites of spleen deficiency and phlegm dampness syndrome were screened out by UHPLC-ESI-MS/MS(the levels of 39 metabolites increased significantly and the levels of 48 metabolites decreased significantly), with the representatives of glycine, L-isoleucine, N-acetyl-L-tyrosine, xanthine, hypoxanthine, and trigonelline.The differential metabolites were mainly enriched in the pathways of steroid hormone biosynthesis, linoleic acid metabolism, and purine metabolism.This study distinguished and revealed the characteristic metabolic pattern of spleen deficiency and phlegm dampness syndrome by metabolomics.The preliminary construction of the OPLS-DA model provides an objective basis for the differentiation of spleen deficiency and phlegm dampness syndrome in traditional Chinese medi-cine(TCM), as well as ideas and methods for exploring the biological basis of TCM syndrome from the molecular level and the overall level.

Hypoglycemic and Hepatoprotective Effects of Dried and Rice-Fried Psidium guajava Leaves in Diabetic Rats.

Psidium guajava leaves (PGL) have been long used as an adjuvant therapy for diabetics. The present study evaluated the in vivo hypoglycemic and hepatoprotective effects of dried and the rice-fried PGL decoctions (PGLD and RPGLD). Our results indicated that both PGLD and RPGLD could significantly decrease the contents of fasting blood glucose (FBG) and haemoglobin A1c (HbA1c) in diabetic rats. Compared with the HFD/STZ (high-fat diet with streptozotocin) group, the PGLD and RPGLD-treated diabetic rats showed different degrees of recovery against the liver pathological changes. The upregulated expressions of glucokinase (GK), glucose transporter 2 (GLUT2), insulin growth factor-1 (IGF-1), insulin receptor substrate-1 (IRS-1), and insulin receptor substrate-2 (IRS-2) in PGLD and RPGLD-treated groups were observed. In general, RPGLD exhibited a much better antidiabetic effect than PGLD, which was further verified by the comprehensive evaluation with the TOPSIS method. Besides, HPLC (high-performance liquid chromatography) and UPLC-MS/MS (ultra-performance liquid chromatography-tandem mass spectrometry) analyses revealed that the contents of the primary constituents (ellagic acid, hyperoside, isoquercitroside, reynoutrin, guaijaverin, auicularin, and quercetin) in RPGLD increased obviously compared with PGLD. These results shed new light on the antidiabetic potential and mechanism of PGL, as well as the "higher efficacy" of the rice-fried processing method in traditional Chinese medicine.

Meta-Analysis on the Chinese Herbal Formula Xiaoer-Feike Granules as a Complementary Therapy for Children With Acute Lower Respiratory Infections.

BACKGROUND: Over the past five years the Chinese herbal formula (CHF) medicine, Xiaoer-Feike granules (XFG), has become a widely used adjuvant therapy for acute lower respiratory infections (ALRI). Considering the rapid popularization and application of XFG, and the lack of systematic evidence evaluating its effectiveness and safety in treating ALRI, it is necessary to conduct a meta-analysis to determine its benefits for patients. METHODS: This study systematically identified randomized controlled trials (RCTs) of XFG treatments for ALRI through July 2019 using four English-databases (PubMed, Cochrane Library, Ovid, and Web of Science) and four Chinese-databases (Sino-med database, China National Knowledge Infrastructure (CNKI), VIP database, and the WANFANG database). We then performed a quality assessment and data analysis with Review Manager 5.3.5 and Stata 15.1. RESULTS: Twenty-one RCTs involving 3425 patients were randomly divided into an XFG group and a conventional medicine (CM) group. The results showed that the clinical efficacy rate (CER) of the XFG group was significantly higher than that of the CM group (RR=1.17, 95% CI =1.13-1.22, P< 0.00001). In comparison with the CM group, the XFG group had strikingly shortened: resolution time of cough (RTC) (MD = -1.92; 95% CI =-2.33, -1.51, P<0.00001); resolution time of rale (RTR) (MD = -1.68; 95% CI =-2.27, -1.10, P<0.00001); resolution time of fever (RTF) (MD = -1.46; 95% CI =-1.92, -1.00, P<0.00001); resolution time of inflammatory lesions (RTIL) (MD = -2.43, 95% CI =-2.94, -1.93, P< 0.00001); and hospital stays (HS) (MD = -2.26, 95% CI =-3.03, -1.49, P< 0.00001). At the cellular and molecular level, the CD4, CD8, CD4/CD8, IL-6, TNF-α, and CRP levels were significantly improved when CM was complemented with XFG. In addition, no significant difference was observed between the XFG and CM groups in terms of the adverse events (AE) (RR =0.97, 95% CI= 0.61-1.54, P= 0.89). CONCLUSIONS: The findings of this meta-analysis support the use of XFG in the treatment of ALRI. However, these results should be treated with caution due to the significant heterogeneity and publication bias of existing data. Further well-designed and high-quality RCTs are needed to interrogate the efficacy and safety of XFG.

Meta-Analysis on the Efficacy and Safety of Hyperbaric Oxygen as Adjunctive Therapy for Vascular Dementia.

Background: Vascular dementia (VD) is a common type of disease in the elderly. Numerous clinical trials have suggested that hyperbaric oxygen is an effective and safe complementary therapy for aging-related disorders. However, there is no reliable systematic evidence regarding hyperbaric oxygen therapy (HBOT) for the treatment of VD. Therefore, we performed a meta-analysis to evaluate the clinical efficacy and safety of HBOT in treating VD. Methods: We methodically retrieved the clinical studies from eight databases (PubMed, Cochrane Library, Embase, Web of Science, Sino-Med, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), and WanFang) from their inception to November 2018. RevMan 5.3.5 was used for quality assessment and data analysis. Stata 15.1 was employed for publication bias detection and sensitivity analysis. Results: Twenty-five randomized clinical trials (RCTs) involving 1,954 patients met our inclusion criteria. These articles researched the HBOT + oxiracetam + conventional therapy (CT) vs. oxiracetam + CT (n = 13), HBOT + butylphthalide +CT vs. butylphthalide + CT (n = 5), HBOT + donepezil + CT vs. donepezil + CT (n = 4), HBOT + nicergoline + CT vs. nicergoline + CT (n = 2) and HBOT + CT vs. CT (n = 1). The results indicated that additional HBOT strikingly improved the Mini-Mental State Examination (MMSE) (MD = 4.00; 95% CI = 3.28-4.73; P < 0.00001), activities of daily living (ADL) (MD = -5.91; 95% CI = -6.45, -5.36; P < 0.00001) and ADL by Barthel index (BADL) (MD = 13.86; 95% CI = 5.63-22.10; P = 0.001) and increased the total efficacy rate (TEF) (OR = 4.84, 95% CI = 3.19-7.33, P < 0.00001). The adverse events rates were not statistically significant between the HBOT and CT groups (OR = 0.85, 95% CI = 0.26-2.78, P = 0.79). Conclusion: In view of the effectiveness and safety of HBOT, the present meta-analysis suggested that HBOT can be recommended as an effective and safe complementary therapy for the treatment of VD. Protocol Registration: PROSPERO (ID: CRD42019117178). Available online at: http://www.crd.york.ac.uk/PROSPERO/display_record.asp?ID=CRD42019117178.

A new abietane diterpenoid glycoside from ajuga ovalifolia var. calantha.

A new abietane diterpenoid glycoside, ajugaside B (1), along with three known compounds (2-4), were isolated from the whole plants of Ajuga ovalifolia var. calantha. The structure of the new compound (1) was elucidated by means of spectroscopic analyses (HRESIMS, IR, NMR and ECD). All of the isolated compounds were evaluated for their antitumor activities against MGC803, MCF-7, A549, HT29 and HepG2 cell lines. Compounds 3-4 showed moderate cytotoxicity against all tested cell lines with IC50 values of 1.8-7.3 µM.

The Increase of Flavonoids in Pericarpium Citri Reticulatae (PCR) Induced by Fungi Promotes the Increase of Antioxidant Activity.

There are thousands of traditional Chinese medicines in China, and they all have to be stored for a period of time for the reason of market price or other factors. But some traditional Chinese medicines especially need to be stored longer, and the clinical efficacy will be better. The cause and mechanism of this phenomenon have attracted much attention in recent years. In this study, we analyze the reason of "the longer storage period the better" of Pericarpium Citri Reticulatae (PCR); method of microscopic and molecular identification was used to identify the fungi separated and purified from PCR. The HPLC and UV spectrophotometry methods were used to determine the contents of flavonoids in PCR. The isolated fungi were inoculated into sterile samples to screen the fungi closely related to the change of flavonoids. The results indicate that the strain of Aspergillus niger could obviously promote the contents of flavonoids, and it could also increase the antioxidant effect of PCR. In conclusion, this study explains the reason of "the longer storage period the better" of PCR from the perspective of microbe, proving the beneficial effect of microorganism on the surface of PCR.

[GC-MS combined with AMDIS and Kováts retention index to investigate dynamic change rules of volatile components from Atractylodis Macrocephalae Rhizoma with different stir-baking degrees].

To investigate the dynamic change rules of volatile components from Atractylodis Macrocephalae Rhizoma with different stir-baking degrees (from slight stir-baking, stir-baking to yellow, stir-baking to brown, to stir-baking to scorch). In the present experiment, the Atractylodis Macrocephalae Rhizoma samples with different stir-baking degrees were collected at different processing time points. The contents of volatile oil in various samples were determined by steam distillation method, and the volatile compounds were extracted by using static headspace sampling method. Gas chromatography-mass spectrography (GC-MS) and automated mass spectral deconrolution and identification system (AMDIS) were combined with Kováts retention index to analyze the chemical constituents of the volatile compounds. The results showed that with the deepening of the stir-baking degree, the content of volatile oil was decreased step by step in 4 phases, and both the compositions and contents of volatile components from Atractylodis Macrocephalae Rhizoma showed significant changes. The results showed that the dynamic change rules of volatile components from Atractylodis Macrocephalae Rhizoma in the process of stir-baking were closely related to the processing degree; in addition, Atractylodis Macrocephalae Rhizoma and honey bran had adsorption on each other. These results can provide a scientific basis for elucidating the stir-baking (with bran) mechanism of Atractylodis Macrocephalae Rhizoma.

Dissipation Pattern, Processing Factors, and Safety Evaluation for Dimethoate and Its Metabolite (Omethoate) in Tea (Camellia Sinensis).

Residue levels of dimethoate and its oxon metabolite (omethoate) during tea planting, manufacturing, and brewing were investigated using a modified QuEChERS sample preparation and gas chromatography. Dissipation of dimethoate and its metabolite in tea plantation followed the first-order kinetic with a half-life of 1.08-1.27 d. Tea manufacturing has positive effects on dimethoate dissipation. Processing factors of dimethoate are in the range of 2.11-2.41 and 1.41-1.70 during green tea and black tea manufacturing, respectively. Omethoate underwent generation as well as dissipation during tea manufacturing. Sum of dimethoate and omethoate led to a large portion of 80.5-84.9% transferring into tea infusion. Results of safety evaluation indicated that omethoate could bring higher human health risk than dimethoate due to its higher hazard quotient by drinking tea. These results would provide information for the establishment of maximum residue limit and instruction for the application of dimethoate formulation on tea crop.

[Study of reason of "the older, the better" of Pericarpium Citri Reticulatae (PCR) which contributed increase of flavonoids].

This study is to explore the reason of "the older, the better" of PCR and itsincrease of flavonoids. We identified the fun- gus isolated from the PCR using microscopic and molecular identification. HPLC method was used to determine the content of 4 fla- vonoids and to clarifythe regularity of them; UV spectrophotometry method was used to determine the total content of flavonoids; reverse thinking was applied to screen the fungus that have close relation to the change of flavonoids. Finally, we have isolated and identified 25 fungusfrom the PCR, which belong to 2 genus and 4 species, including pencillium commune, P. minioluteeum, P. citrinum, Aspergillus flavus and A. niger. The content of flavonoids was increased in the mildew PCR due to A. niger and other fungus. Therefore, "the ol- der, the better" of PCR had its scientific reason that the increase of flavonoids had a close relation of the metabolic activity of A. niger and other fungus.

[Deconvolution of overlapped peaks in total ion chromatogram of essential oil from citri reticulatae pericarpium viride by automated mass spectral deconvolution & identification system].

This experiment shows how to use the automated mass spectral deconvolution & identification system (AMDIS) to deconvolve the overlapped peaks in the total ion chromatogram (TIC) of volatile oil from Chineses materia medica (CMM). The essential oil was obtained by steam distillation. Its TIC was gotten by GC-MS, and the superimposed peaks in TIC were deconvolved by AMDIS. First, AMDIS can detect the number of components in TIC through the run function. Then, by analyzing the extracted spectrum of corresponding scan point of detected component and the original spectrum of this scan point, and their counterparts' spectra in the referred MS Library, researchers can ascertain the component's structure accurately or deny some compounds, which don't exist in nature. Furthermore, through examining the changeability of characteristic fragment ion peaks of identified compounds, the previous outcome can be affirmed again. The result demonstrated that AMDIS could efficiently deconvolve the overlapped peaks in TIC by taking out the spectrum of matching scan point of discerned component, which led to exact identification of the component's structure.

[Determination of sulfur dioxide in traditional Chinese medicine by derivative fluorometry].

OBJECTIVE: To establish a derivative fluorometry method for the determination of sulfur dioxide residues in traditional Chinese medicine. METHOD: The optimal derivation condition was established. The fluorescence intensity was detected at excitation wavelength of 321 nm, and emission wavelength of 384 nm. RESULT: A linear relationship was obtained between the fluorescence intensity and the addition of reference substance in the range of 0.999 7-17.99 nmol with a correlation coeffient of 0.999 9, and the average recovery was 102.3% with RSD 4.6%. CONCLUSION: This method is simple and sensitive with quick and correct result. It can provide a reference for the determination of sulfur dioxide residues in traditional Chinese medicine.

[Chemical variability of essential oils in peels collected in different time from Citrus reticulata 'Ponkan'].

OBJECTIVE: To study the chemical variation of essential oils in peels collected from Citrus reticulata' Ponkan' in different time. METHODS: The volatile oils were extracted by steam distillation and analyzed by GC-MS. The oil components were identified by their mass spectra and Kovats retention indices (RIs) and quantified by the area normalization method. RESULTS: A total of 68 compounds were identified in 6 samples harvested in different time, and the total contents of identified compounds in each sample were from 94.27% to 97.12%. The highest content compounds were d-limonene, gamma-terpinene and linalool representing 56.862%-67.728%, 9.298%-11.081% and 4.792%-7.893%, respectively. There was little difference in the chemical components between different samples, but a great variation in quantitation between the former 2 samples and the latter 4 samples. CONCLUSION: The chemical variability of essential oils from 6 Citrus reticulata 'Ponkan' peel samples presents a regularity.

[Contents comparision of atractylode I, II, III in rhizoma atractylodis macrocephalae and the processed with soils].

OBJECTIVE: To research the effect of processing with soils on the contents of Atractylode I, II, III of Rhizoma Atractylodis Macrocephalae. METHODS: Contents of Atractylode I, II, III were determined by HPLC. Hypersil-ODS C18 (200 mm x 4.6 mm, 5 microm) column was used; The mobile phase consisted of acetonitrile-water; The flow rate was 1.0 mL/min; The detection wavelength was 220 nm and 276 nm,the column temperature was 30 degrees C. RESULTS: All the indexes of Atractylode I, II, III peak were qualified; The result of methodological study was met the requirement of HPLC; The average contents of Atractylode I , II , II in raw Rhizoma Atractylodis Macrocephalae were 0.4365, 0.2878, 0.4140 mg/g, the soil-processed Rhizoma Atractylodis Macrocephalae were 0.5503, 0.3013, 0.8403 mg/g, the processed Rhizoma Atractylodis Macrocephalae were 0.5386, 0.2958, 0.7399 mg/g. CONCLUSION: The content of Atractylode II is no significant difference among the raw Rhizoma Atractylodis Macrocephalae and the Processed. The contents of both Atractylode I and III increased after processed. However, there is no significant difference between the soil-processed and the processed Rhizoma Atractylodis Macrocephalae with nothing.