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To prepare the mimetic exosomes and co-delivery proteins and nucleic acids, and achieve efficient and safe co-delivery of multi-component drugs, an optimized formulation was designed by modifying a polylactic acid-glycolic acid copolymer (PLGA) matrix with a cationic lipid excipient dioleyl trimethylammonium propane (DOTAP), and a PLGA/DOTAP nanoparticles packaged protein and nucleic acid was prepared by double emulsion method, and the outermost membrane structure prepared by reverse phase evaporation method and consists of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and membrane proteins. The structure of the mimetic exosomes is formed by ultrasonic dispersion and extrusion, and analyzed its characteristics and nature of the transfer effect. The size of mimetic exosomes was about 156.13 nm, with negative charge (-18.23 ± 0.57 mV), and it could efficiently co-transfer protein and siRNA, and siRNA could effectively inhibit the expression of target gene Trim28. The mimetic exosomes simulate the structure of exosomes and achieve safe and efficient co-delivery of multi-component drugs.
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This research is to establish an HPLC method for determination of geniposidic acid, genipin-1-β-D-gentiobioside, geniposide, p-trans-coumaroylgenipin gentiobioside, chlorogenic acid, crocin-Ⅰ, crocin-Ⅱ and crocin-Ⅲ in Gardeniae Fructus at different harvest time. The detection wavelength was 238, 320 and 440 nm. Principal component analysis(PCA), correlation analysis, regression analysis and partial least squares(PLS) analysis were used to explore the relationship of color and content of eight components in Gardeniae Fructus. The result showed that the trend of the eight components in Gardeniae Fructus at harvest time in different three years was varied similarly. According to the variation of eight components at different harvest time, the mature and immaturate Gardeniae Fructus were discriminated. The content of crocin-Ⅰwas correlated positively with a~* of color significance. The redder color of Gardeniae Fructus showed the higher value of a~* and content of crocin-Ⅰ, indicating the better quality of Gardeniae Fructus. This method provided reference for justifying the color and quality of Gardeniae Fructus and scientific evidence for "assessing quality by distinguishing color".
Subject(s)
Chlorogenic Acid , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fruit , GardeniaABSTRACT
Objective::To compare the effect of different medicinal parts of Nandina domestica in reducing toxicity of anti-tumor drug arsenic trioxide, so as to provide the scientific basis for its further development and application. Method::Chronic arsenic trioxide poisoning model was used in this paper. Totally 56 SD rats were randomly divided into normal group, model group (arsenolite 40 mg·kg-1), sodium 2, 3-dimercapto-1-propanesulfonate group (sodium 2, 3-dimercapto-1-propanesulfonate 25 mg·kg-1+ arsenolite 40 mg·kg-1), Nandinae Radix group (Nandinae Radix 20 g·kg-1+ arsenolite 40 mg·kg-1), Nandinae Caulis group (Nandinae Caulis 20 mg·kg-1+ arsenolite 40 mg·kg-1), Nandinae Folium group (Nandinae Folium 20 g·kg-1+ arsenolite 40 mg·kg-1), and Nandinae Fructus group (Nandinae Fructus 20 g·kg-1+ arsenolite 40 mg·kg-1). The intragastric administration lasted for 10 days. After the last administration, urine was collected within 24 hours, serum, kidney and liver tissue samples were collected after operation, and serum creatinine (SCr) and urine creatinine (UCr) levels were measured, in order to calculate endogenous creatinine clearance rate (CCr). At the same time, the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in liver and kidney of rats in each group were detected. Some kidneys and livers were fixed with formaldehyde, and the histopathological changes were observed under microscope after hematoxylin-eosin staining. Result::Compared with the model group, the rats in combination group of Nandinae Radix, Nandinae Caulis, Nandinae Fructus have a heavier body mass (P<0.01), the kidney coefficient was lower (P<0.01), the levels of UCr and CCr were significantly increased (P<0.01), the content of MDA in renal tissue was decreased significantly (P < 0.01), the level of MDA in liver tissue was decreased (P < 0.05), whereas the activities of SOD and CAT were significantly increased (P<0.01), the pathological damage of liver and kidney was alleviated. There was no significant difference in the activity of SOD in the liver between the Nandinae Folium combination group and the model group, but the changes of the other indexes were consistent with those of the above three groups. Conclusion::Nandinae Radix, Nandinae Caulis, Nandinae Fructus have significant protective effects on liver and kidney toxicity induced by arsenic trioxide oxidative stress, and Nandinae Folium was the least effective among them.
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Objective::To study the effect of total alkaloids of Nandina domestica in attenuating arsenic trioxide, and analyze chemical constituents of alkaloids extract of Nandina, in order to provide the theoretical basis for studying the effect of N. domestica in attenuating arsenic trioxide with alkaloids extract of N. domestica as effective fraction. Method::The model of acute liver injury induced by arsenic trioxide was used to compare the effects on heart and liver functions of mice between arsenic trioxide alone and total alkaloids of N. domestica combined with arsenic trioxide. The detoxification of total alkaloids on arsenic trioxide was evaluated based on biochemical parameters and pathological report. Peakview (Version1.2, AB SCIEX) software was used to process the mass spectrometry data of total alkaloids of N. domestica. The structures of determined compounds were identified by molecular weight of compound (molecular formula), secondary fragments, chromatographic peak retention and literature information. Result::Among biochemical indicators, creatine kinase(CK), lactate dehydrogenase(LDH), blood urea nitrogen(BUN), malondialdehyde(MDA), alanine aminotransferase(ALT) and amino transferase of aspartate(AST) of the arsenic trioxide group were increased, while elimination rates of Na+ -K+ -adenosine triphosphate(ATP), superoxide dismutase(SOD), catalase(CAT) and serum creatinine(SCr) were decreased, compared with those of the combination group. CK and LDH of the alkaloids extract group were more obviously increased than those of the N. domestica extract group, but with no remarkable difference. In histomorphometric examination, edema of mouse heart cells was improved, and some kidney and liver damages in rats were alleviated. Totally 25 alkaloids of alkaloid extract were identified. Among them, 18 were known, and 7 were unknown, including 3 structural types, in which apomorphine alkaloids were mostly. Conclusion::Heart, kidney and liver damage degrees of the combination group were significantly alleviated compared with the arsenic trioxide group. The total alkaloids fraction extracted and purified have a significant attenuation in arsenic trioxide toxicity. The detoxification of total alkaloids extract was equal to that of N. domestica extract. Furthermore, apomorphine alkaloids, such as nantenine and domesticine, can be used as index components to establish a quality control method for total alkaloids.
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Objective::Histomorphological study of the Nandinae Radix, Nandinae Caulis, Nandinae Folium, Nandinae Fructus were conducted to provide the basis for the identification of its authenticity and falsehood. Method::The origin and macroscopic identification were used to describe the plant morphology and the appearance characteristics of all medicinal parts. The microscopic characteristics of the medicinal parts, such as roots, stems, leaves, flowers, fruits, were observed and photographed by paraffin section and powder preparation techniques. Result::It was found that the morphological characteristics of the original plant were consistent with the descriptions in herbaceous books. There was no pith on the cross-section of roots. In the transverse section of stems, there were intermittent circular fiber bundles in the cortex and a cap-shaped fibrous bundle in the inner part of the xylem. In the transverse section of the leaves, the palisade tissue was wider and the fiber bundles around the main vascular bundles formed a ring. In the transverse section of petiole, the fiber bundles were arranged intermittently into rings. In the transverse section of fruit, multi-layered sclereids formed a ring in the mesocarp. The powder characteristics of root and stem mainly contained crystalliferous sclereids. There were crystal sheath fibers and stomatal infinitive in the leaf powder, and pollen grains in the flower powder, with 3-hole grooves, obvious reticulate engraving pattern in the outer wall and more reticulate cells. There were a large number of branched sclereids and calcium oxalate square crystals in the fruit powder. Conclusion::The above-mentioned morphological and microscopic features have identification significance, and provide scientific basis for the authenticity identification, the quality standard and the utilization of resources of Nandina domestica.
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Objective:To investigate the effective substance of the choleretic effect of Gardeniae Fructus，and analyze the relationships between its choleretic effect and the HPLC fingerprint chromatogram. Method:HPLC method was applied to establish the fingerprint chromatography of 8 batches of Gardeniae Fructus at different harvest periods. The flow，the content of bile acid，bilirubin and cholesterol in bile were tested，and then the principal component analysis was used to comprehensively evaluate the total choleretic effects of Gardeniae Fructus. After the relationships between the relative peak area of the common peaks and the choleretic effects were explored using grey relationship analysis method，the spectrum-effect relationship of Gardeniae Fructus was established. Result:The order of the contribution of the chemical components to the choleretic effect at the common peaks was as follow（r>0.8）：P9>P14>P26>P4>P30>P6>P1>P10>P5>P24. Among all peaks，the full wavelength scanning results implied that the peaks 9，14 and 4 might be iridoids, and the peaks 26，30 and 24 might be crocins. By comparing with the standard substances，the peak 9 was finally identified as geniposide. Conclusion:The choleretic effect of Gardeniae Fructus may be the results of multiple components and pathways，and the main components in Gardeniae Fructus with the choleretic effect was from geniposide. In conclusion，these results provide a reference for investigating the material basis of choleretic effect of Gardeniae Fructus.
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At present, it is generally believed that the paracrine effect of stem cells in the repair of myocardial injury is one of the important ways for stem cell therapy. Exosomes are phospholipid bilayer-enclosed nanovesicles that secreted by cells under physiological and pathological conditions. Cargo loaded into exosomes including protein, lipids and nucleic acids can be delivered to recipient cells. Therefore, exosomes are recognized as important mediators for intercellular communication. It has been suggested that exosomes from stem cells (eg. embryonic stem cells, induced pluripotent stem cells, cardiac progenitor cells, mesenchymal stem cells and cardiosphere-derived cells) have protective effects against heart injury. In this review, we summarized recent research progresses on stem cell-derived exosomes in myocardial injury, including the therapeutic effects and mechanism.
Subject(s)
Humans , Cell Communication , Exosomes , Physiology , Heart Injuries , Induced Pluripotent Stem Cells , Cell Biology , Mesenchymal Stem Cells , Cell BiologyABSTRACT
Aim To determine the effect of exosomes from lipopolysaccharide-treated human bone marrow mesenchymal stem cells on proportion of Ly6Chigh and Ly6Clow monocytes/macrophages in inflammatory micro- environment. Methods BMSCs were obtained by gra-dient centrifugation, identified and then treated with li-popolysaccharide for 48 h. The exosomes were purified from conditional medium with or without LPS treatment and identified by CD63 protein using Western blot and transmission electron microscope. The diameters and concentration were detected by Nanoparticle Trafficking Analysis ( NTA ) . The monocytes/macrophages were sorted from bone marrow of the mice by magnetic beads. Cells were co-cultured with exosomes for 24 hours, and then treated with LPS for 48 hours. The proportion of Ly6C monocytes/macrophages was detec-ted by flow cytometry. Inflammatory cytokines in cell supernatant were investigated using ELISA. Results BMSCs surface markers CD44, CD90 were positively detected, but CD34, CD45 were not expressed. BM-SCs presented adipogenic differentiation ability. Exo-somes were positively expressing CD63 protein, and NTA showed that the diameters of exosomes were up to (82.4 ± 3.7 ) nm. BMSCs stimulated by LPS pro- duced more exosomes ( P < 0.01 ) . Exosomes from BMSCs with or without LPS treatment could increase the ratio of Ly6Chigh monocytes (P<0.01) and down-regulate the ratio of Ly6Chigh macrophages (P<0.05), and the effect of LPS treated-exosomes was more signif-icant than untreated-exosomes (P<0.05). Moreover, the concentration of IL-6 was also elevated under exo-somes treatment ( P <0.05 ) . Conclusions Human bone marrow mesenchymal stem cells-derived exosomes contribute to the regulation of Ly6Chigh monocytes/mac-rophages, indicating that they could be involved in the therapeutic treatment of inflammatory diseases.
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Objective To establish an HPLC fingerprint method of Gardeniae Fructus which could provide the basis for its authenticity identification and quality evaluation. Methods HPLC method was adopted to construct the fingerprint of Gardeniae Fructus and its main confused species (G. jasminoides f. logicarpa and G. jasminoides var. grandiflora). HPLC-ESI-MS was applied to identify the common peaks. The cluster analysis (CA) and principal component analysis (PCA) were used to classify the samples. Results The HPLC fingerprint of Gardeniae Fructus was obtained with 17 common peaks, and 15 common peaks were identified using HPLC-ESI-MS. A total of 22 batches of the fruits of Gardenia were classified to Gardeniae Fructus, G. jasminoides Ellis f. logicarpa, and G. jasminoides var. grandiflora by CA and PCA. Conclusion The establishment of HPLC fingerprint of Gardeniae Fructus and the application of chemical pattern recognition can provide scientific basis for authenticity identification and quality control of Gardeniae Fructus.
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Through the textual research, resource investigation, literature reviews (including Flora of China, municipal Flora, pharmacopoeia of China and municipal drug standards) and identification of commercial drugs on Cuscutae Semen, it was found the species described in the herbal textual was Cuscuta chinensis, with good quality from Shandong and Henan Province. The identification of commodities showed the majority drugs were from C. australis, varied from the ancient herbal textuals .Mordern literature reviews indicate that it was necessary to strengthen the research on Cuscutae Semen from C. australis, C. chinensis and C. japonica because of their differences in resources, macroscopical and microscopical characters, while wrong descriptions in some literatures. It was suggested that the two species (C. australis and C. chinensis) should be separated in pharmacopoeia of China. The study provides scientific basis for the development and utilization of Cuscutae Semen.
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To set standards for histomorphological studies on Lysimachia fortunei, an efficacious and widely applied folk medicine in this study, in order to develop its resources. Its species were identified by observing plant morphology and herbs appearance characters, preparing slices with routine methods and defining structural characters. According to the results of morphologic observation, leaves, stamen and pistil of this plant were different from the descriptions in Flora of China. The whole herb can be used in medicines, mainly including rhizomes, stems and leaves. According to the findings in the first study on microscopic structures, its rhizomes, stems and leaves were characteristic and worth identifying. The transaction tissue structures of rhizomes and stems were under developed and contained endodermis, secretory structures; Stems had sclerenchymata of different shapes of sclereids; Leaves were bifacial and had vascular bundles under midribs, which were surrounded by parenchymal sheathes. On the surface of leaves, stomata, glandular hairs and keratin lines were morphologically different in upper and lower epidermis. The herbal power had glandular hairs, sclereids and vessels. In conclusion, herbs of L. fortunei can be identified by the above histomorphological characteristics, which lays a foundation for further development and application of L. fortunei.
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Medicine, Traditional , Plant Leaves , Plant Stems , Plants, Medicinal , PrimulaceaeABSTRACT
<p><b>OBJECTIVE</b>To clarify the origin and provide pharmacognostical evidences for the leaves of 5 species in Chloranthus.</p><p><b>METHOD</b>Histological observation and microscopic identification through different slice-making techniques were applied to the research.</p><p><b>RESULT</b>There were subtle differences between the histological characteristics. In microscopical identification, the different structures of vascular bundles in veins were observed, appendages and non-glandular hairs were distinct.</p><p><b>CONCLUSION</b>The method can be used to distinguish the features of 5 species in Chloranthus. This article offers information for the further research and exploitation of Chloranthus.</p>
Subject(s)
Magnoliopsida , Pharmacognosy , Plant Epidermis , Plant Leaves , Plant Vascular Bundle , Plants, Medicinal , Species SpecificityABSTRACT
<p><b>OBJECTIVE</b>To investigate the expressions of human telomerase reverse transcriptase (h-TERT), c-myc, and proliferating cell nuclear antigen (PCNA) in chronic viral hepatitis (CVH), liver cirrhosis and primary hepatocellular carcinoma (HCC) and understand their possible role in liver carcinogenesis.</p><p><b>METHODS</b>Totally 157 liver disease specimens were collected, including 56 CVH, 52 liver cirrhosis and 49 primary HCC specimens. In situ hybridization was performed on these specimens to examine the expressions of h-TRET and c-myc mRNA, and immunohistochemistry carried out for PCNA detection, with the cell apoptosis detected with in situ ending labeling.</p><p><b>RESULTS</b>In the CVH, liver cirrhosis and primary HCC specimens, h-TERT expression was detected at the frequencies of 11/56 (19.6%), 43/52 (82.7%) and 44/47 (93.6%), c-myc expression at 7/56 (12.5%), 21/52 (40.4%) and 26/47 (55.3%), with apoptotic index of (27.3-/+4.7)%, (16.5-/+2.6)% and (8.7-/+1.3)% and PCNA expression rate of (17.1-/+2.9)%, (49.3-/+7.8)% and (62.5-/+9.1)%, respectively. Correlations among h-TERT, c-myc, and PCNA expressions and the apoptotic index were not found in the examined tissues (P>0.05).</p><p><b>CONCLUSION</b>Liver carcinogenesis may involve increased h-TERT, c-myc, and PCNA expressions and suppressed cell apoptosis.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Apoptosis , Carcinoma, Hepatocellular , Genetics , Metabolism , Pathology , Cell Transformation, Neoplastic , Hepatitis B, Chronic , Genetics , Metabolism , Pathology , Immunohistochemistry , Liver Cirrhosis , Genetics , Metabolism , Pathology , Liver Neoplasms , Genetics , Metabolism , Pathology , Proliferating Cell Nuclear Antigen , Proto-Oncogene Proteins c-myc , Genetics , RNA, Messenger , Genetics , Metabolism , Telomerase , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the mechanism of anti-tumor activity of Acanthopanax gracilistylus extract (Age).</p><p><b>METHODS</b>The tumor cells proliferation was detected by using (3H)-TdR incorporation method, and the effects of Age on cell cycle of tumor cells, retinoblastoma (Rb) protein and cyclin-dependent kinases (Cdk) were analyzed by flow cytometry and Western blotting assay, respectively.</p><p><b>RESULTS</b>It was indicated by cytoactivity test in vitro that Age only had effect in inhibiting the proliferation of tumor cells, it couldn't lead to death of cells. Under action of Age, the proliferation of tumor cells was halted at G0/G1 stage of cell cycle, and showed no direct cytotoxic effect by Age. Age could induce lowering of the expression of Rb, Cdk2 and Cdk4, cause halt of tumor cell proliferation.</p><p><b>CONCLUSION</b>The tumor inhibitory effect of Age is realized by way of regulating the activity of cell cycle controlling enzymes to suspend the proliferation of tumor cells.</p>