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Supercritical fluid extraction (SFE) employing carbon dioxide (SC-CO2) is an efficient method to extract bioactive compounds from agro-forest wastes. These compounds maintain and/or improve food nutrition, safety, freshness, taste, and health and are employed as natural functional food components. To highlight the potential of this technology, we focus on the following current advances: (I) parameters affecting solubility in SFE (pressure, temperature, SC-CO2 flow rate, extraction time, and co-solvents); (II) extraction spectra and yield obtained according to proportion and composition of co-solvents; (III) extract bioactivity for functional food production. Fatty acids, monoterpenes, sesquiterpenes, diterpenoids, and low-polarity phenolic acids and triterpenoids were extracted using SFE without a co-solvent. High-polarity phenolic acids and flavonoids, tannins, carotenoids, and alkaloids were only extracted with the help of co-solvents. Using a co-solvent significantly improved the triterpenoid, flavonoid, and phenolic acid yield with a medium polarity.
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Chromatography, Supercritical Fluid , Functional Food , Chromatography, Supercritical Fluid/methods , Forests , Functional Food/analysis , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Waste Products/analysisABSTRACT
Objective:To investigate the mid-term clinical outcomes of selective column arthrodesis based on the three-column theory in the treatment of malunion of Lisfranc injury.Methods:The 28 patients with malunion of Lisfranc injury were analyzed retrospectively who had been treated by selective column arthrodesis at Department of Orthopedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine from January 2011 to January 2020.They were 18 males and 10 females, with an average age of 37.2 years(from 18 to 65 years). Twelve left and 16 right sides were affected. According to Myerson's three-column classification, one case was medial column injury (type A), 4 ones middle column injury (type B), 7 ones medial plus middle columns injury and 16 ones three-column injury. Medial column arthrodesis was conducted in 7, middle column arthrodesis in 4 and medial plus middle columns arthrodesis in 17. The American Orthopaedic Foot and Ankle Society (AOFAS) midfoot score and visual analogue scale (VAS) were compared between preoperation and the last follow-up to evaluate the improvements in foot function and pain. The operation-related complications were recorded.Results:All patients were followed up for an average of 35.6 months (from 18 to 60 months). The AOFAS midfoot score increased from 43.1±4.1 at pre-operation to 84.1± 7.4 at the last follow-up and the VAS score decreased from 5.7±1.3 at pre-operation to 2.0±0.9 at the last follow-up (both P<0.001). The wounds healed in 28 patients, 3 of whom had postoperative wound exudation but responded to dressing change. There were no such complications as injury to the deep peroneal nerve or deep venous thrombosis. The internal fixation was removed in 5 patients at about one year after arthrodesis. Conclusion:Selective column arthrodesis based on the three-column theory can result in satisfactory med-term clinical outcomes in the treatment of malunion of Lisfranc injury.
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Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.
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<p><b>INTRODUCTION</b>Diabetes mellitus induces microangiopathic changes that lead to endothelial dysfunction. This study investigated the effect of Xiaokening, a type of Chinese compound medicine, on the mesenteric arteriolar endothelial cell function of diabetic rats and its underlying mechanism.</p><p><b>METHODS</b>Diabetes mellitus was induced in rat models via intraperitoneal injection of 60 mg/kg streptozotocin and observed over three weeks. Mesenteric arterioles, which were isolated in a cannulated and pressurised state, were incubated with intravascular injections of 1, 3 or 5 g/L Xiaokening for 24, 48 or 72 hours. The effects of Xiaokening on the release of nitric oxide (NO) on the mesenteric arterioles were detected under shear stress of 1, 10 and 20 dyn/cm(2). Biochemical methods were used to determine the activities of superoxide dismutase (SOD) and xanthine oxidase (XO). The expressions of endothelial NO synthase (eNOS), SOD and XO in the mesenteric arterioles were assessed using Western blot.</p><p><b>RESULTS</b>Compared to normal rat arterioles, less NO was released in the mesenteric arterioles of diabetic rats. Xiaokening was found to have a concentration- and time-dependent effect on NO release; when the shear stress was increased, there was a gradual increase in the release of NO. Compared to normal arterioles, the expression of eNOS in the mesenteric arterioles of diabetic rats was lower. Incubation with Xiaokening increased SOD activity and expression, and decreased XO activity and expression in the mesenteric arterioles of the diabetic rats.</p><p><b>CONCLUSION</b>Xiaokening was able to significantly increase NO release and improve the endothelial function of mesenteric arterioles through antioxidative mechanisms.</p>
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Animals , Male , Rats , Antioxidants , Chemistry , Arterioles , Diabetes Mellitus, Experimental , Drug Therapy , Dose-Response Relationship, Drug , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Injections, Intraperitoneal , Nitric Oxide , Metabolism , Nitric Oxide Synthase Type III , Metabolism , Oxygen , Metabolism , Rats, Sprague-Dawley , Streptozocin , Superoxide Dismutase , Metabolism , Xanthine Oxidase , MetabolismABSTRACT
Purpose To observe the effects of midkine ( MK) on human breast cancer cell line MDA-MB-231 angiogenesis in vitro, and to explore its mechanism. Method shRNA interference was performed to silence the expression of MK in MDA-MB-231 cells, and Western blot was used to identify the expression of MK and EPCR. After MK and EPCR knockdown, or treated with anti protease-activated receptor 1 (PAR1) antibody, the culture medium of MDA-MB-231 cells were collected and the conditioned medium were pre-pared. Human umbilical vein endothelial cells ( HUVECs) were cultured with conditioned medium, and the endothelial cells prolifera-tion was detected by CCK-8 assay, cell migration was detected by transwell method, vasculogenic activity was assessed by Matrigel-based tube formation assay. Results After knockdown of MK, the protein level of EPCR was decreased in MDA-MB-231 cells. Com-pared with control, knockdown of MK and EPCR decreased the proliferation, migration and angiogenesis ability of HUVECs significant-ly (P<0. 05), and the effect of EPCR knockdown group was stronger than MK knockdown group (P<0. 05). After treated with anti-PAR1 antibody, the proliferation, migration and angiogenesis ability of HUVECs were decreased compared with control and EPCR knockdown group (P<0. 05). Conclusion MK promotes human breast cancer cell line MDA-MB-231 angiogenesis through EPCR /PAR1 signaling pathway in vitro.
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Purpose To observe the effect of MK on proliferation, migration and angiogenesis of human umbilical vein endothelial HU-VECs cells and to explore the role of MK in tumor angiogenesis. Methods siRNA targeting MK was used to downregulate MK expres-sion in human breast cancer cell line MDA-MB-231. Then, the experiment was divided into three groups: the untreated group, the empty vector transfection group and MK gene interference group. CCK-8 assay was used to detect the endothelial cells proliferation, tr-answell method was for detection of endothelial cell migration numbers, and matrigel in vitro small tube formation assay was used to sur-vey the state of tube formation. Expression of MK in siRNA transfection was identified by RT-PCR and Western blot. Results The MK gene interference group showed lower cell proliferation activity, less number of migration of cells and tube formation than the other two groups (P<0. 05). Conclusion Low-expressed MK in human breast cancer cell line MDA-MB-231 can inhibit proliferation, mi-gration and angiogenesis of endothelial cells, which shows that MK may play an important role in tumor angiogenesis.
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Objective To explore clinical effect of reconstruction for chronic Achilles tendon rupture of Kuwada IV type with flexor hallucis longus (FHL) harvested using a minimally invasive technique.Methods The data of 35 patients with chronic Achilles tendon rupture of Kuwada IV type was retrospectively analyzed who were treated by FHL which was harvested using a minimally invasive technique from July 2006 to June 2011.There were 21 males and 14 females,with the age from 23 to 71 years (average,42.1 years).All patients were unilateral injury.MRI showed Achilles tendon rupture fissures 6.0-9.2 cm.The local appearance and function recovery on postoperation was observed,and all patients were assessed with the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Leppilahti Achilles tendon repair score.Results Thirty-two patients were followed up for 18 to 72 months,with an average of 33.2 months.Except for 1 patient whose wound healed after six weeks through resuture immediately for the wound dehiscence occurred in the ten days,other patients' wound healed smoothly.The average of AOFAS ankle-hindfoot score had increased from 51.92±7.08 preoperatively to 92.56±6.71 postoperatively.Leppilahti Achilles tendon repair score had increased from 72.56±7.43 preoperatively to 92.58±5.1 postoperatively.Twenty-seven cases were excellent,good in 3,and fair in 2,with the total excellent and good rate 93.8% (30/32).No case of the sural nerve and tibial nerve injury,plantar painful scar,plantar outside nerve injury.MRI of Achilles tendon showed even signal without signal of tear and cystic degeneration.Conclusion Reconstruction for chronic Achilles tendon rupture of Kuwada IV type with FHL harvested using a minimally invasive technique offers a desirable outcome in rapid postoperative recovery,high strength in tenodesis,fewer complications.
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<p><b>OBJECTIVE</b>To establish a HPLC-DAD model for the simultaneous determination of two selaginellins (selaginellin and selaginellin B) and four biflavones (amentoflavone, sequoiaflavone, hinokiflavone and isocryptomerin) contained in 10 batches of Selaginella tamariscina and 12 batches of S. pulvinata produced in different areas.</p><p><b>METHOD</b>The analysis was performed on a Waters Cosmosil C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril-0.1% formic acid as mobile phase in a linear gradient mode. The detection wavelength was set at 280, 337 nm. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was 30 degrees C.</p><p><b>RESULT</b>The six active constituents showed significant different in content. Amentoflavone in S. tamariscina contains (5. 628-9. 184 mg x g(-1)) is more than that contained in S. pulvinata (0.823-7.131 mg x g(-1)), while selaginellin in S. pulvinata (0.123-0.593 mg x g(-1)) is more than that contained in S. tamariscina (0.067-0.133 mg x g(-1)). All the calibration curves showed good linear correlation coefficients (r > 0.9997) over the wide test ranges.</p><p><b>CONCLUSION</b>The developed HPLC-DAD method is simple, sensitive and accurate and has the good repeatability in separation, which is available for the quality control of S. tamariscina and S. pulvinata.</p>
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Biflavonoids , Chemistry , Biphenyl Compounds , Chemistry , Chromatography, High Pressure Liquid , Methods , Cyclohexanones , Chemistry , Flavones , Chemistry , Flavonoids , Chemistry , Selaginellaceae , ChemistryABSTRACT
AIM: The study aimed to verify the effects of arsenic trioxide in combination with aspirin on angiogenesis of human gastric cancer xenografts in nude mice. METHODS: Experiments were performed at the Laboratory of Pathology, Xuzhou Medical College from September 2006 to July 2007. A total of 36 male BALB/C-nu/nu nude mice aged 4-6 weeks, weighting 18-22 g were selected. Human gastric cancer cell strain SGC-7901 were purchased from Shanghai Institute of Biology of Chinese Academy of Sciences. Thirty-six nude mouse models of cell strain SGC-7901 lateral armpit subcutaneously transplantation tumor were established and randomized into a control group, a aspirin group, a arsenic trioxide group and a combination group with nine in each group. Seven days after inoculation, nude mice were intraperitoneally administrated with drugs. Nude mice in the control group were treated with sterile saline (0.2 mL/d); Mice in the aspirin group were administrated with aspirin 0.3 g/(kg?d); Mice in the arsenic trioxide group received 2.5 mg/(kg?d); Mice in the combination group were administrated with 2.5 mg/(kg?d)plus aspirin 0.3 g/(kg?d), successively for 14 days. Immunohistochemical method (SP kit) was used to detect the expression of cycloxygenase-2 (COX-2) and CD34, and count microvessel density (MVD). Western-blotting was used to detect the expression of IKK?, I?B, nuclear factor (NF)-?B and vascular endothelial growth factor (VEGF). RESULTS: A total of 27 male BALB/C-nu/nu nude mice were included in the final results. Seven days after inoculation, 1 mm tumor was formed on the back of 31 nude mice. One mouse respectively died in the arsenic trioxide group and the combination group. One mouse died and one loss in the aspirin group. Compared to the aspirin group, arsenic trioxide group and control group, expressions of IKK?, NF-?B and VEGF decreased in the combination group (P
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Purpose:To study the roles of cyclin D1 and cyclin E in non small cell lung cancer(NSCLC) and the relationship between cyclin D1 and cyclin E.Methods:The expression of cyclin D1 and cyclin E in 87 NSCLC were studied immunohistochemically, PCNA was detected immunohistochemically to estimate the proliferative index(PI), the relationship between cyclin D1, cyclin E, PI, clinicopathological and prognostic factors were analysed.Results:Of the 87 NSCLC, 44 8%(39/87) were positive for cyclin D1 and 48.3%(42/87) for cyclin E, the PI of the cyclin D1 positive group was significantly higher than the cyclin D1 negative group( P 〈0.05).Significant differences were shown in the diameter of tumor, lymph node metastasis rate and survival rate between the positive and negative group of cyclin D1( P 〈0.01,0.05,0.01), and there were significant differences in lymph node metastasis, stages and survival rate between the positive and negative group of cyclin E( P 〈0.05,0.05,0.01). The expression of cyclin E in the cyclin D1 positive group was significantly higher than that of the cyclin D1 negative group( P 〈0.05), and the PI, diameter of tumor, lymph node metastasis rate in the coexpression group of cyclin D1 and cyclin E were significantly higher than that of the non coexpression group( P 〈0.05), while the coexpression group had significantly lower survival rate than that of the non coexpression group( P 〈0.05).Conclusions:Cyclin D1 and cyclin E are involved in NSCLC, but their effects are different: cyclin D1 is an important factor in the regulation of proliferation, cyclin E is related to the progression of NSCLC, cyclin D1 can promote the expression of cyclin E, while cyclin E can perhaps intensify the effect of cyclin D1, thus there could exist synergism between cyclin D1 and cyclin E.
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Objective: To study the expression of cyclin D1 and E in lung cancer. Methods: The immunohistochemical technique was used to detect the expression of cyclin D1 and E in 68 cases of non small cell lung cancer(NSCLC) and 21 cases of small cell lung cancer(SCLC). Results: The rate of cyclin D1 protein positive expression was 48.5% in NSCLC, 4.8% in SCLC,showing significant difference( P
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Objective To explore the effects and relationship of tert-butyl hydroperoxide (t-BHP) and hypoxia in transient ischemia-like attack (TIA-like) in Kunming mice.Methods Fifty-six mice were randomly divided into normal saline group (group A), t-BHP group (group B), t-BHP plus hypoxia group (group C) and model group (group D). Fourteen mice in group D were injected t-BHP (0.11 mol/L, 10 ml/kg) through tail vein with an interval of 24 hrs to induce TIA-like attack. The average time of TIA-like was ( 3.7? 1.1) days, and this time was used as experimental evidence to collect blood sample in other groups. Hemorheological indexes before TIA-like attack were observed in different experimental conditions.Results Compared with group A, the whole blood viscosity at shear rate 100S-1, 1S-1 and Fibrinogen were significantly increased in group B and group C (all P
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Objective: To develop a method to determine Astragaloside IV in spray-dry of Radix Astragali by HPLC-ELSD. Methods: ELSD was used to determine directly Astragaloside IV in spray-dry of Radix Astragali on Elite-OSD column, using CH 3CN-H 2O(36∶64) as a mobile phase. The flow rate was 1.0 ml/min. The temperature of drift tube for ELSD was 105 ?C and the flow rate of N 2 was 2.84 ml/min. Results: The recovery of the added sample was 91.68% and RSD 1.64%. Soluble amylum was helpful for spray-dry but it can influence was determination of Astragaloside IV. Conclusion: The method is accurate and can be used in the determination of Astragaloside IV in spray-dry of Radix Astragali. Better adjuvant should be found.