A Novel Strategy for Screening Active Components in Cistanche tubulosa Based on Spectrum-Effect Relationship Analysis and Network Pharmacology.

Cistanche tubulosa (Schenk) R. Wight is a valuable herbal medicine in China. The study aimed to explore the potential mechanisms of C. tubulosa on antioxidant activity using spectrum-effect relationship and network pharmacology and the possibilities of utilizing herbal dregs. In this work, different extracts of C. tubulosa, including herbal materials, water extracts, and herbal residues, were evaluated using high-performance liquid chromatography (HPLC) technology. In addition, the antioxidant activities were estimated in vitro, including 2, 2-diphenyl-1-picrylhydrazyl; superoxide anion; and hydroxyl radical scavenging assays. The spectrum-effect relationships between the HPLC fingerprints and the biological capabilities were analyzed via partial least squares regression, bivariate correlation analysis, and redundancy analysis. Furthermore, network pharmacology was used to predict potential mechanisms of C. tubulosa in the treatment of antioxidant-related diseases. According to the results, eleven common peaks were shared by different extracts. Geniposidic acid, echinacoside, verbascoside, tubuloside A, and isoacteoside were quantified and compared among different forms of C. tubulosa. The spectrum-effect relationship study indicated that peak A 6 might be the most decisive component among the three forms. Based on network pharmacology, there were 159 target genes shared by active components and antioxidant-related diseases. Targets related to antioxidant activity and relevant pathways were discussed. Our results provide a theoretical basis for recycling the herbal residues and the potential mechanisms of C. tubulosa in the treatment of antioxidant-related diseases.

The localization of the alkaloids in Coptis chinensis rhizome by time-of-flight secondary ion mass spectrometry.

Background: Understanding the spatial distribution of active compounds can effectively evaluate the quality of decoction pieces of traditional Chinese medicine (TCM). Traditional methods are economical and practical but lack chemical information on the original distribution. Time-of-flight secondary ion mass spectrometry (TOF-SIMS), with the advantage of non-destructive detection of samples, can directly analyze the distribution of chemical compounds on the surface of various samples. Methods: In this study, TOF-SIMS image analysis technology was used to detect TCM for the first time. Taking Coptis rhizome (CR) as an example, a commonly used TCM, the distribution of the compounds in the cross-section of CR was studied. Meanwhile, ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLCQQQ-MS/MS) was used to verify the results of TOF-SIMS. Results: The distribution of nine active compounds: berberine, epiberberine, coptisine, palmatine, columbamine, jatrorrhizine, tetrahydricheilanthifolinium, and oxyberberine, was well imaged in the cross-section of CR by TOF-SIMS. The content of berberine and epiberberine was the highest; Palmatine distribution in the pith was more than that in other parts; Oxyberberine was mainly concentrated in the cork and xylem rays. Normalization analysis showed contents of these compounds increased along with the growth years. The result was consistent with UPLC-QQQ-MS/MS. Conclusion: The TOF-SIMS method can display the spatial distribution status of the active compounds of herbs, providing a basis for selecting the medicine site with non-destructive and fast detection.

Metabolic Insight Into the Neuroprotective Effect of Tao-He-Cheng-Qi (THCQ) Decoction on ICH Rats Using Untargeted Metabolomics.

Tao-He-Cheng-Qi decoction (THCQ) is an effective traditional Chinese medicine used to treat intracerebral hemorrhage (ICH). This study was performed to investigate the possible neuroprotective effect of THCQ decoction on secondary brain damage in rats with intracerebral hemorrhage and to elucidate the potential mechanism based on a metabolomics approach. Sprague-Dawley (SD) rats were randomly divided into five groups: the sham group, collagenase-induced ICH model group, THCQ low-dose (THCQ-L)-treated group, THCQ moderate-dose (THCQ-M)-treated group and THCQ high-dose (THCQ-H)-treated group. Following 3 days of treatment, behavioral changes and histopathological lesions in the brain were estimated. Untargeted metabolomics analysis with multivariate statistics was performed by using ultrahigh-performance liquid chromatography-mass spectrometry (UPLC-Q-Exactive Orbitrap MS). THCQ treatment at two dosages (5.64 and 11.27 g/kg·d) remarkably improved behavior (p < 0.05), brain water content (BMC) and hemorheology (p < 0.05) and improved brain nerve tissue pathology and inflammatory infiltration in ICH rats. Moreover, a metabolomic analysis demonstrated that the serum metabolic profiles of ICH patients were significantly different between the sham group and the ICH-induced model group. Twenty-seven biomarkers were identified that potentially predict the clinical benefits of THCQ decoction. Of these, 4 biomarkers were found to be THCQ-H group-specific, while others were shared between two clusters. These metabolites are mainly involved in amino acid metabolism and glutamate-mediated cell excitotoxicity, lipid metabolism-mediated oxidative stress, and mitochondrial dysfunction caused by energy metabolism disorders. In addition, a correlation analysis showed that the behavioral scores, brain water content and hemorheology were correlated with levels of serum metabolites derived from amino acid and lipid metabolism. In conclusion, the results indicate that THCQ decoction significantly attenuates ICH-induced secondary brain injury, which could be mediated by improving metabolic disorders in cerebral hemorrhage rats.

Corrigendum to "Selection of quality markers of Jasminum amplexicaule based on its anti-diarrheal and anti-inflammatory activities: Effect-target affiliation-traceability-pharmacokinetics strategy" [Chinese Herbal Medicines 11 (2019) 379-386].

[This corrects the article DOI: 10.1016/j.chmed.2019.08.002.].

[Quality evaluation of Bolbostemmatis Rhizoma by UPLC fingerprint combined with QAMS].

The present study was performed to establish the UPLC fingerprints of Bolbostemmatis Rhizoma and determine the contents of three saponins by quantitative analysis of multi-components by single marker(QAMS), and provide basis for quality evaluation of Bolbostemmatis Rhizoma. The analysis was carried out on an analytical column of Waters Cortecs T3(2.1 mm×100 mm,1.6 µm)with gradient elution by acetonitrile-0.1% phosphoric acid solution, at a flow rate of 0.3 mL·min~(-1). The detection wavelength was 203 nm, the column temperature was 30 ℃ and the injection volume was 1 µL. The UPLC fingerprints of Bolbostemmatis Rhizoma were established and evaluated by similarity calculation, cluster analysis and principal component analysis. The relative calibration factors of toberoside B and toberoside C were determined with toberoside A as internal reference. The content was calculated by relative calibration factors to develop a method of QAMS. Comparing the results of QAMS with those of ESM, the accuracy and feasibility of one-eva-luation and multi-evaluation can be determined. RESULTS:: showed that the fingerprints of 19 batches of Bolbostemmatis Rhizoma have four common peaks with similarities ranging from 0.754 to 1.000. Cluster analysis and principal component analysis classified 19 batches of Bolbostemmatis Rhizoma into three categories, which was consistent with the similarity evaluation results. The relative deviation between the content of tubeicosides B and C in 19 batches of Bolbostemmatis Rhizoma determined by QAMS and ESM is less than 5.0%, indicating that there was no significant difference between the two methods. Therefore, the UPLC fingerprints combined with QAMS and similarity evaluation can be effectively used to evaluate the quality of Bolbostemmatis Rhizoma.

[Multi-component quantitation of artificial cow-bezoar and study on its markers of quality difference].

An HPLC-ELSD method for quantitation of GCANa, CA, HDCA, CDCA and DCA in artificial cow-bezoar was developed in this study. The chromatographic separation was performed using a Wondasil CC18(4.6 mmχ250 mm，5 µm) column at a column temperature of 25 °C and liquid flow-rate of 1.0 mL·min⁻¹. Acetonitrile and 0.2% formic acid solution were used as mobile phase with a linear gradient and injection volume was 10 µL. An ELSD was used with a nitrogen flow-rate of 2.3 L·h⁻¹at a drift tube temperature of 110 °C. Some chemometric methods were applied in data procession and analysis for exploration of the markers of quality difference. The accurate and simple method is suitable for the quality evaluation and the quality control of artificial cow-bezoar.

Trichiconlides CF, four new limonoids with 1,2-seco phragmalin-type carbon skeleton from the fruits of Trichilia connaroides.

Phytochemical investigation of the fruits from Trichilia connaroides led to the isolation of four rare 1,2-seco phragmalin-type limonoids (1-4) with C-7/28 δ-lactone ring along with two known triterpenoids (5-6). An oxygen bridge between C-1 and C-2 formed an rare 9-oxa-tricyclo[3.3.2.17,10]undecane-2-ene moiety in structure of 1 and 2, and 3-4 possessed a bicyclo[5.2.14,10]decane skeleton. Their structures were established based on extensive spectroscopic methods and their absolute configurations of 1 were confirmed by TDDFT/ECD calculation and comparative analysis. Compounds 5-6 showed moderate inhibitory effects on Hela tumor cell lines with inhibition ratio values of 83.8% and 87.1% at concentration 40µM, respectively.

[The analysis of multivariate image and chemometrics in TLC fingerprinting of artificial cow-bezoar].

A method of thin-layer fingerprinting chromatogram of artificial cow-bezoar was established with the developing solvent consisting of cyclohexane, ethyl acetate, acetic acid and methanol (2∶7∶1∶2), and 10% sulfuric acid ethanol solution sprayed as colour-developing agent. After heated at 105 ℃, TLC was recorded as an image in ultraviolet light at 366 nm which was converted into grayscale. By the gray value extracted from the grayscale, the multivariate data obtained from TLC of samples could be analyzed by chemometric method. The results indicated that samples from different manufacturers could be distinguished by this method and some specific bands were found out. All in one, this simple and practical method was suitable for the evaluation of quality difference.

[Formulation Optimization of Zuojin Floating-Bioadhesive Pellets by Central Composite Design-Response Surface Methodology].

OBJECTIVE: To optimize the formulation of Zuojin floating-bioadhesive pellets by the central composite design-response surface methodology (CCD-RSM). METHODS: In the formulation design using CCD-RSM, independent variables were the amounts of sodium bicarbonate (X1), HPMC (X2) and MCC (X3) as factors. Small pills roundness, 12 h floating rate and the percentages of in vitro cumulative releases at 2,6 and 12 h were dependent variables. Multilinear and quadratic model were used to estimate the relationship between the dependent and the independent variables. According to best model, the contour plots and RSM were drawn, and the optional formulation was selected. According to the optional formulation,the pellets were prepared and validated. RESULTS: The quadratic was the best fitting mode. Small pills roundness was 15.04 °. 12 h floating rate was 75.07%. Percentages of in vitro cumulation release at 2,6 and 12 h of the option formulation pellets was 27.01%, 70.00% and 84.61%, respectively. The actual value was close to the predicted value. Deviation was less than 5%. CONCLUSION: Quadratic model was preferred in the optimization of formulation due to the statistical confidence. The multi-objective simultaneous optimization of Zuojin floating pellet formulation can be achieved by the central composite design-response surface methodology.

[Evaluation of the immediate effect of acupuncture on cervical spondylosis of vertebral artery type based on orthogonal design].

OBJECTIVE: To analyze the differences in the immediate effect of acupuncture on cervical spondylosis of vertebral artery type among three factors: needling technique, acupoint selection and time of needle retaining so as to option the best therapeutic program. METHODS: Thirty-two cases were observed and randomly divided into 8 groups, 4 cases in each one. The orthogonal design of three factors and two levels was adopted. There were needling technique (electroacupuncture, reinforcing and reducing method), acupoint selection [C4-C7 Jiaji (EX-B 2) and three needles of neck: Tianzhu (BL 10), Jingbailao (EX-LHN 15), Dazhu (BL 11)] and time of needle retaining (5 min, 15 min). L8 (2(7)) orthogonal design table was arranged in the trial. The changes in mean velocity (Vm) of vertebral artery (VA) and basilar artery (BA) were observed before and after acupuncture. RESULTS: The immediate effect of VA-BA blood flow was the most significant after electroacupuncture at C4-C7 Jiaji (EX-B 2), with continuous wave for 5 min. This method and acupoint selection greatly influenced the therapeutic results (both P<0.05). CONCLUSION: The optimized therapeutic program of the immediate effect of acupuncture for cervical spondylosis of vertebral artery type is electroacupuncture + C4-C7 Jiaji (EX-B 2) + 5 min. The importance of different factors for the immediate effect in acupuncture treatment of cervical spondylosis of vertebral artery type is: acupoint selection > needling technique > time of needle retaining.

[Genetic analysis of family constellation for cochlear implant recipients].

GJB2, SLC26A4 (PDS) and mitochondrial DNA (mtDNA) have been associated with sensorineural hearing loss. In the present study, the clinical, genetic and molecular analysis of 14 cochlear implant recipients and their parents was studied from April 2006 to September 2007. Of the 14 subjects, 35.7% had gene mutations; 28.6% had homozygous GJB2 235delC mutation, whose parents carried heterozygous GJB2 235delC mutation; and 7.1% had mtDNA A1555G mutation, whose mother carried mtDNA A1555G mutation too. There was no SLC26A4 (PDS) mutation. These results strongly suggested that the mutation in GJB2 gene was a major cause of deafness in cochlear implant recipients and the mutation of mtDNA A1555G was another important cause. Genetic test of hot-spots and analysis of family constellation can offer an accurate genetic counseling to deaf family and reduce the incidence of hearing loss.

[Determination of amentoflavone in 11 species of Selaginella medicinal material by HPLC].

OBJECTIVE: To determine the content of amentoflavone in 11 species of Selaginella medicinal materials. METHODS: HPLC was used with kromasil (250 x 4. 6 mm, 5 microm) column. The mobile phase consisted of methanol and phosphate buffer. The gradient condition was 0-->28 min with methanol 50%-->58%, and 30-->35 min with methanol 58%-->70%. The column temperature was 40 degrees C. The flow rate was 1. 0 ml/min and the detection wavelength at 330 nm. RESULTS: Amentoflavone showed a good linearity in the range of 153 - 767 ng. The average recovery was 97.03% and RSD was 0. 80% (n = 5). CONCLUSION: The method was rapid, accurate and can be used for quality control of Selaginella medicinal materials.