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OBJECTIVE@#To elucidate the underlying mechanism of Panax notoginseng saponin (PNS) on gastric epithelial cell injury and barrier dysfunction induced by dual antiplatelet (DA).@*METHODS@#Human gastric mucosal epithelial cell (GES-1) was cultured and divided into 4 groups: a control, a DA, a PNS+DA and a LY294002+PNS+DA group. GES-1 apoptosis was detected by flow cytometry, cell permeability were detected using Transwell, level of prostaglandins E2 (PGE2), 6-keto-prostaglandin F1α (6-keto-PGF1α) and vascular endothelial growth factor (VEGF) in supernatant were measured by enzyme linked immunosorbent assay (ELISA), expression of phosphatidylinositide 3-kinase (PI3K), phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), glycogen synthase kinase-3β (GSK-3β) and Ras homolog gene family member A (RhoA) were measured by Western-blot.@*RESULTS@#DA induced apoptosis and hyper-permeability in GES-1, reduced supernatant level of PGE2, 6-keto-PGF1α and VEGF (P<0.05). Addition of PNS reduced the apoptosis of GES-1 caused by DA, restored the concentration of PGE2, 6-keto-PGF1α and VEGF (P<0.05). In addition, PNS attenuated the alteration of COX-1 and COX-2 expression induced by DA, up-regulated p-PI3K/p-Akt, down-regulated RhoA and GSK-3β. LY294002 mitigated the effects of PNS on cell apoptosis, cell permeability, VEGF concentration, and expression of RhoA and GSK-3β significantly.@*CONCLUSIONS@#PNS attenuates the suppression on COX/PG pathway from DA, alleviates DA-induced GES-1 apoptosis and barrier dysfunction through PI3K/Akt/ VEGF-GSK-3β-RhoA network pathway.
Subject(s)
Humans , Cyclooxygenase 1 , Epithelial Cells/metabolism , Glycogen Synthase Kinase 3 beta , Panax notoginseng , Phosphatidylinositol 3-Kinases/metabolism , Platelet Aggregation Inhibitors , Proto-Oncogene Proteins c-akt/metabolism , Saponins/pharmacology , Vascular Endothelial Growth Factor A , rhoA GTP-Binding ProteinABSTRACT
Objective:To investigate and compare the fingerprints of different polarity fractions (petroleumether,chloroform,ethyl acetate,n-butanol,water) of fresh Gardeniae Fructus with different fruit shapes,and further understand the content difference and distribution of its chemical composition. Method:Gardeniae Fructus was reflux extracted by water in order to obtain the water extract; water extract 0.1 g and dissolved with 50 mL water,then it was extracted by petroleum ether,chloroform,ethyl acetate and n-butanol in turn in order to obtain the different extraction phases and the water phase. Each phase was condensed to extractum. Finally,the samples were analyzed by high performance liquid chromatography (HPLC) fingerprints and the similarity evaluation software was used for data analysis. Result:Fingerprint of chloroform fraction of water extract in gardenia from different habitats can be used to distinguish Gardeniae Fructus from Fujian,Henan and Jiangxi. The differences between the water extract of Gardeniae Fructus from Fujian and those of Henan and Jiangxi were mainly manifested in the petroleum ether fraction, and the fat-soluble components of Gardeniae Fructus were more than those of Henan and Jiangxi. The differences between the water extract of Gardeniae Fructus from Henan and those of Fujian and Jiangxi were mainly manifested in the ethyl acetate fraction,and the content of iridoid glycosides was significantly higher than that in Fujian and Henan. The differences between the water extract of gardenia from Jiangxi and those of Fujian and Henan were mainly manifested in the n-butanol fraction,organic acid peak C1 not detected. The fingerprint of chloroform fraction of water extract in Gardeniae Fructus can be used to distinguish Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs from Fujian and Henan,and the contents of all components of Gardeniae Fructus of seven ribs were more than those in Gardeniae Fructus of six ribs. The fingerprint of petroleum ether fraction of water extract in Gardeniae Fructus can be used to distinguish Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs from Jiangxi. The Z3 peak of Gardeniae Fructus of six ribs from Henan was obviously higher than that of Gardeniae Fructus of seven ribs. The contents of all components on chloroform and ethyl acetate fractions of water extract in Gardeniae Fructus of seven ribs were significantly higher than those of Gardeniae Fructus of six ribs. Conclusion:There are significant differences on chemical constituents and content among Gardeniae Fructus from Fujian,Henan and Jiangxi. The main difference of fingerprint between Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs is the peak height.
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Objective: To determine the content of index components in different parts of Gardenia jasminoides (pericarp, seeds, whiskers), study the fingerprint, and compare the contents and compositions differences of different parts of G. jasminoides, in order to provide the theoretical basis for different efficacies of G. jasminoides pericarp and seeds, explore the exploitation and utilization values of G. jasminoides whiskers, and avoid waste of gardenia medicinal resources. Method: The contents of geniposide and crocetin Ⅰ was were determined by HPLC, the content of total iridoid glycosides was determined by ultraviolet spectrophotometry, and three index components in different parts of G. jasminoides were analyzed. HPLC fingerprints of different parts of G. jasminoides were collected, the common pattern of HPLC fingerprints of different parts of G. jasminoides of different origins and with different processing methods was established, and the similarity evaluation software was used for data analysis; comparative analysis on fingerprints of different parts of G. jasminoides was conducted. Result: Content change of index components in G. jasminoides pericarp and seeds from Henan, Fujian and Jiangxi were the same. Content of geniposide:Fujian > Henan > Jiangxi, the contents of three components in G. jasminoides pericarp from Fujian were much higher than those from Henan and Jiangxi, the contents of crocetin Ⅰ and total iridoid glycosides:Fujian > Jiangxi > Henan, the contents of total iridoid glycosides from Fujian, Jiangxi were much higher than those from Henan. The order of three index components in G. jasminoides whiskers from different origins from high to low, the content of geniposide and crocetin Ⅰ was Fujian > Jiangxi and Henan, the content of total iridoid glycosides was Fujian > Jiangxi > Henan.In the same part, there were 22 common peaks in the fingerprints of G. jasminoides pericarp, except for S13-S15, the similarity of other samples were more than 0.9;the fingerprints of G. jasminoides seeds had 22 common peaks, except for S22-S30, the similarities of other samples were more than 0.9;the fingerprints of G. jasminoides whiskers had 16 common peaks, except for S7-S9, the similarities of other samples were more than 0.9.In different parts, the fingerprints of G. jasminoides whiskers were significant different from those of pericarp and seeds, the number of peaks in G. jasminoides whiskers reduced, the order of height of peaks 2, 3, 5 of G. jasminoides from high to low were whiskers > gardenia > seeds. There was not peak X in the seeds, the height of peak X of gardenia in whiskers was higher than that in pericarp, except for the peak 17, the height of all peaks in seeds were higher than that in whiskers. Conclusion: There are significant differences in the contents of index components in G. jasminoides pericarp and seeds. The content of total glycosides in gardenia is high, suggesting that it can be used to extract total iridoid glycosides. The fingerprints can reflect the content difference and species distribution of different parts of G. jasminoides, so as to provide theoretical support for the studies for pharmacodynamic material basis of G. jasminoides and the scientificity and rationality of the separate application of G. jasminoides pericarp and seeds.
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@#AIM: To determine the effect of obstructive sleep apnea syndrome(OSAS)on visual field and retinal nerve fiber layer(RNFL)thickness.<p>METHODS: This study consisted of 72 eyes of 72 OSAS patients who were enrolled as OSAS group, and compared with 74 eyes of 74 age-matched physical examination personnels who were enrolled as control group. Visual field sensitivities(VFS)were explored with Humphrey perimeter. Peripapillary RNFL thickness was measured by stratus optical coherence tomography(OCT). VFS and Peripapillary RNFL were divided into upper, lower, temporal and nasal regions. According to apnea-hypopnea index(AHI)scores monitored by polysomnography(PSG), VFS and peripapillary RNFL thickness in 29 patients with mild OSAS, 25 patients with moderate OSAS and 20 patients with severe OSAS were compared with those in control group.<p>RESULTS: There was no significant difference in VFS between OSAS group and control group in upper, lower, temporal and nasal regions(<i>P</i>>0.05). RNFL of nasal region in OSAS group, especially in severe OSAS patients, was significantly lower than that in control group(<i>P</i>=0.047). Pearson correlation analysis showed that there was negative correlation between RNFL and OSAS severity in nasal region(<i>r</i>= -0.9998, <i>P</i>=0.0138).<p>CONCLUSION: Severe OSAS may lead to nasal RNFL thickness reduction, and the change of RNFL thickness may be used as one of the indicators to assess the severity of OSAS.
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To investigate the effect of different initial processing methods on the quality of Gardenia and determine the best cooking time in gardenia processing through the determination of index components content. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia were determined before storage, six months after storage and one year after storage. During storage, the contents of geniposide, crocetin Ⅰ and total iridoid glycosides in directly dried Gardenia were 1.68%, 0.45% and 6.45% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different steaming time were 1.34%-0.5%, 0.28%-0.06% and 6.09%-1.59% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different boiling time (adding alum)were 1.42%-0.41%, 0.35%-0.07% and 6.40%-1.65% respectively. The direct drying of Gardenia samples could not achieve the function of killing enzyme and protecting glycosides. The enzymes from degradation of the index components were basically destroyed after steaming time of 13 min or boiling (adding alum) time of 8 min, achieving the function of killing enzyme and protecting glycosides.
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<p><b>OBJECTIVE</b>To investigate the dosage-efficacy/toxicity relationship of prepared rhubarb, in order to explore the bidirectional effects in hepatoprotection and hepatotoxicity of prepared rhubarb and the objective authenticity for attenuating toxicity by processing.</p><p><b>METHOD</b>Normal and pathological animals were adopted simultaneous to investigate the effect of total extracts from prepared rhubarb within a high dose range (2.0, 5.4, 14.7, 40.0 g x kg(-1) x d(-1)) on normal state, biochemical index and histopathology of experimental animals. The factor analytic approach was used to analyze the dosage-efficacy/toxicity relationship of prepared rhubarb.</p><p><b>RESULT</b>The factor analytic approach was used to extract two common factors from the nine biochemical indexes. The firs common factor was mainly dominated by HA, LN and TGF-β1, and could be explained as fibrotic factors. The second common factor was mainly dominated by ALT, AST and ALP, and could be explained as cellular factor. The results of the factor analysis suggested that prepared rhubarb showed significant bidirectional effects in hepatoprotection and hepatotoxicity, which could protect liver in CC14 injured chronic hepatic injury, but had a certain hepatotoxic effect to normal animals. The pathological examination showed consistent results with the factor analysis. Under comparable dosages, prepared rhubarb showed a stronger liver protecting effect than crude rhubarb, with a lower toxicity.</p><p><b>CONCLUSION</b>Although prepared rhubarb has a certain hepatotoxic effect to normal animals, it has also a significant therapeutic effect to animals with liver injury. The results proved the symptom-based prescription theory and the scientificity of the symptom-based medication. The symptom-based prescription theory is important to correctly realize the dosage-efficacy/toxicity relationship of traditional Chinese medicines and guide the symptom-based medication.</p>
Subject(s)
Animals , Female , Male , Rats , Dose-Response Relationship, Drug , Drug Prescriptions , Drugs, Chinese Herbal , Pharmacology , Toxicity , Fibrosis , Liver , Metabolism , Pathology , Rats, Sprague-Dawley , Rheum , Chemistry , Toxicity TestsABSTRACT
<p><b>OBJECTIVE</b>To optimize the conditions of purifying the total alkaloids in Aconitum szechenyianum with macroporous adsorption resin, and compare the content of total alkaloids and aconitine in A. szechenyianum from different producing areas, in order to provide basis for further studies.</p><p><b>METHOD</b>The orthogonal experiment method was adopted for optimizing the conditions of purifying the total alkaloids in A. szechenyianum with macroporous adsorption resin. The content of total alkaloids and aconitine were determined by using the titration method. The total alkaloids in A. szechenyianum from different producing areas were purified under optimum processing conditions. Aconitine was determined by HPLC.</p><p><b>RESULT</b>Different processing conditions showed different influences on the purification of total alkaloids. The optimum conditions were resin type HPD-722, ethanol concentration of 80% , and ethanol elution volume of 80 mL x min(-1). The contents of aconitine in A. szechenyianum from different producing areas--Qinghai, Maxianshan, Ningxia and Yongdeng were 0.493 5, 0.883 5, 1.527 8, 1.664 4 mg x g(-1), respectively.</p><p><b>CONCLUSION</b>The optimum processing conditions used in this essay could be used for purifying the total alkaloids and aconitine. A. szechenyianum from Yongdeng and Gansu contains the highest content of aconitine.</p>
Subject(s)
Aconitine , Chemistry , Aconitum , Chemistry , Adsorption , Alkaloids , Chemistry , China , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Porosity , Resins, Synthetic , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To construct tissue engineering bone with bone marrow stromal cell (BMSC) sheets of dogs.</p><p><b>METHODS</b>BMSC were derived from dog bone marrow and cell sheets were prepared with temperature-responsive dishes after the cells were induced by osteogenesis. Allogeneic dogs decalcification bone matrixes (DBM) were prepared. Sixteen dogs were divided into 4 groups. The MSC cell sheets-rhBMP-2-BMSC-DBM were implanted under the left latissimus dorsi myofascial as the experimental side; while the rhBMP-2-BMSC-DBM were implanted in the right side as the control. Ectopic bone formation in vivo was evaluated by histological examination 4, 8, 12, 16 weeks after operation.</p><p><b>RESULTS</b>The osteogenesis in the experimental group was better than that in the control group. New bone area in the experimental side was larger than that in the control group, and the difference was significant (P < 0.05). After 16 weeks, lamellar bone was connected into a film in the experimental group. Haversian system and red bone marrow could be seen.</p><p><b>CONCLUSIONS</b>BMSC cell sheets could promote the bone formation of functional tissue-engineered bone.</p>
Subject(s)
Animals , Dogs , Bone Marrow Cells , Bone Matrix , Bone and Bones , Mesenchymal Stem Cells , Stromal Cells , Tissue EngineeringABSTRACT
<p><b>OBJECTIVE</b>To study the effect of maxillary sinus floor elevation by the Frialit-2 Bone Condenser for implantation.</p><p><b>METHODS</b>11 patients underwent sinus floor lift by The Frialit-2 Bone Condenser and were inserted 14 implants. The time of following up was 10 - 21 months.</p><p><b>RESULTS</b>There were no implant loose or lost, no clinical complaint of maxillary sinus area, and X-ray exam showed well osseointegration.</p><p><b>CONCLUSIONS</b>The Frialit-2 bone condenser can be used for lifting sinus floor, and the sinus elevation without lateral access allows the insertion of implants with no additional surgical stress for the patients.</p>
Subject(s)
Female , Humans , Male , Alveolar Process , General Surgery , Bone Transplantation , Dental Implantation , Methods , Dental Implants, Single-Tooth , Follow-Up Studies , Maxillary Sinus , General SurgeryABSTRACT
Objective:The immunological effects of HCV-DNA vaccine with different adjuvants were detected by ELISPOT in mice.Methods:Female BALB/c mice were primed with naked HCV-DNA, HCV-DNA encapsulated by liposome DDAB/EPC or DC-Chol/DOPE, HCV-DNA mixed with Montanide ISA 720 or aluminum hydroxide, respectively, and boosted twice accordingly in a four-week interval. Cytokine production by splenocytes was assessed by ELISPOT.Results:In most cases, splenocytes from mice vaccinated with DDAB/EPC liposome produced more IFN-?. These splenocytes also have significant higher IL-2 production compared with the other groups. In expansion with NS5b, splenocytes from alum group have significance in IL-4 production compared with other groups. The profile of cytokine production revealed that the INF-? overwhelmed IL-4 in naked DNA, DDAB/EPC, and DC-Chol/DOPE groups while IL-4 surmounted IFN-? in alum and Montanide groups.Conclusion:Encapsulation with liposome DDAB/EPC has the strongest adjuvant effect in inducing Th1 dominated immunity. Alum and Montanide can convert the Th1 nature of DNA vaccine to Th2-biased immunity.