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ObjectiveTo investigate the effect of alcohol extract of Oroxylum indicum （MHD-80） on reducing uric acid （UA） and protecting the kidney in the hyperuricemia （HUA） model in vivo. MethodPotassium oxazine （350 mg·kg-1） and adenine （80 mg·kg-1） were used to construct an HUA model of mice in vivo to evaluate the mechanism related to UA reduction and the protective effect of renal function of MHD-80. Seventy male ICR mice were randomly divided into seven groups， including the normal group， model group， allopurinol group （5 mg·kg-1）， febusotan group （5 mg·kg-1）， and MHD-80 low-， medium-， and high-dose groups （3， 6， 12 mg·kg-1）， with 10 in each group. Except for the normal group， the other groups were given intragastric administration of potassium oxazine and adenine for 14 consecutive days to establish the HUA model. On the 8th to 14th day after modeling， each group was given corresponding drugs by intragastric administration， once a day. 1 h after the last administration， blood was collected from the eyeballs， and kidney and liver tissues of mice were collected. Serum levels of UA， urea nitrogen （BUN）， and creatinine （Cr） and liver activity of xanthine oxidase （XOD） were determined by enzyme colorimetry. Serum contents of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were determined by enzyme-linked immunosorbent assay （ELISA）. Hematoxilin-eosin （HE） staining was used to observe the pathological changes in kidney tissues. The protein expression levels of ATP-binding box transporter G2 （ABCG2） and glucose-facilitating transporter 9 （GLUT9） in kidney tissues were detected by Western blot. ResultIn vivo experiment shows that compared with the normal group， the serum levels of UA， Cr， BUN， inflammatory factors TNF-α， IL-1β， and liver XOD activity in the serum of mice in the model group were significantly increased （P<0.05， P<0.01）， and the expression of GLUT9 in kidney tissues was significantly up-regulated （P<0.05）. ABCG2 protein expression was significantly down-regulated （P<0.05）， and renal injury was obvious. Compared with the model group， the levels of UA， BUN， Cr， TNF-α， IL-1β， and liver XOD activity in the serum of mice in the high-dose group of MHD-80 were decreased to different degrees （P<0.05， P<0.01）， GLUT9 protein expression was significantly down-regulated （P<0.01）， ABCG2 protein expression was significantly up-regulated （P<0.05） in the high-dose group of MHD-80， and the degree of renal injury was reduced. ConclusionMHD-80 has certain uric acid reduction， anti-inflammatory， and anti-renal injury effects， which are related to inhibiting XOD activity and regulating the expression of ABCG2 and GLUT9 uric acid transporter.
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@#Objective To investigate the predictive value of right atrial myocardial fibrosis in the prognosis of isolated tricuspid regurgitation surgery after left heart valve surgery. Methods The patients who underwent tricuspid valvuloplasty by the same operator in Guangdong Provincial People's Hospital from April 2016 to August 2021 due to long-term isolated severe tricuspid regurgitation after left heart valve surgery were included in the study. According to the degree of right atrial myocardial fibrosis, the patients were divided into three groups: a mild group, a moderate group, and a severe group. The clinical data of these patients were compared and analyzed. Results A total of 75 patients were enrolled, including 16 males and 59 females with an average age of 57.0±8.4 years. There were 30 patients in the mild group, 29 patients in the moderate group and 16 patients in the severe group. In terms of the preoperative data, there were statistical differences in cardiac function grade, right atrial diameter, tricuspid incompetence area among the three groups (P<0.05). In terms of the postoperative data, there were statistical differences among the three groups in the cardiopulmonary bypass time, mechanical ventilation time, ICU monitoring time, complication rate and mortality (P<0.05). Further pairwise comparison showed that, compared with the mild group, the severe group had longer mechanical ventilation time (P=0.024), longer ICU monitoring time (P=0.003) and higher incidence of postoperative complications (P=0.024), while the moderate group had no statistical difference in all aspects (P>0.05); compared with the moderate group, the severe group had longer ICU monitoring time (P=0.021) and higher incidence of complications (P=0.006). Conclusion The early outcome of tricuspid valvuloplasty in patients with isolated tricuspid regurgitation after left heart valve surgery with severe right atrial myocardial fibrosis is worse than that in the patients with mild and moderate fibrosis, suggesting that the degree of myocardial fibrosis in the right atrium can be a predictor of the effect of tricuspid regurgitation surgery and a judgement indicator of the surgery timing.
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@#Objective To summarize the early outcomes of totally thoracoscopic minimally invasive aortic valve replacement (AVR) and double valve replacement (DVR). Methods The clinical data of patients who underwent totally thoracoscopic minimally invasive AVR or DVR in Guangdong Provincial People’s Hospital from April 2020 to January 2021 were retrospectively analyzed. The patients were divided into an AVR group and a DVR group according to the surgical method, and the clinical data of the two groups were compared. Results Finally 22 patients were enrolled, including 14 males and 8 females with an average age of 50.0±11.2 years at operation. Eight patients were degenerative disease, 8 were rheumatic heart disease combined with valvular disease, and 6 were bicuspid aortic valve. Out of the 22 patients, 16 underwent AVR alone, and 6 underwent DVR. All patients completed the operation successfully, and there was no death. Perivalvular leakage during surgery occurred in 2 patients. The average cardiopulmonary bypass time was 187.0±39.9 minutes, and aortic cross-clamping time was 117.0 (99.0, 158.0) minutes. Duration of mechanical ventilation and intensive care unit stay was 9.5 (4.8, 18.3) hours and 41.0 (34.0, 64.0) hours, respectively. The volume of chest drainage at the first 24 hours after surgery was 214.0±124.6 mL, and the postoperative hospital stay was 5.5 (4.0, 8.3) days. The cardiopulmonary bypass time and aortic cross-clamping time in the DVR group were longer than those in the AVR group, and the volume of chest drainage at 24 hours after surgery was more than that in the AVR group, with a statistical difference (P<0.05). Echocardiography before hospital discharge showed paravalvular leakage in 1 patient. There was no death during follow-up of 5.9±3.0 months. Conclusion The early outcome of totally thoracoscopic minimally invasive AVR and DVR is satisfactory, and the approach of surgery is worth exploring.
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Objective: To elucidate the anti-inflammatory mechanism of Reduning Injection (RDN) by analyzing the potential biomarkers and metabolic pathways of the carrageenan-induced inflammatory model from the overall metabolic level. Methods: Rat inflammatory model was established by carrageenan. UPLC-Q-TOF/MS was used to detect and analyze changes of endogenous metabolites in the serum and urine of carrageenan-induced inflammatory rats. Combined with multivariate analysis and databases analysis, inflammatory-related potential biomarkers were screened and identified to analyze possible metabolic pathways. The reliability and biological significance of these biomarkers was verified by metabolic network analysis and correlation analysis with pharmacodynamic indicators. Results: A total of 16 potential biomarkers were screened and identified by multivariate analysis and metabolite databases, among which 13 species could be adjusted by RDN. The metabolism pathway analysis revealed that histidine metabolism, sphingolipid metabolism, and tyrosine metabolism were greatly disturbed. Their biomarkers involved urocanic acid, sphingosine, and norepinephrine, all of which showed a callback trend after RDN treatment. The three biomarkers had a certain correlation with some known inflammatory-related small molecules (histamine, arachidonic acid, Leukotriene B4, and PGE
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Objective:To evaluate the efficacy and safety of perioperative rehabilitation approaches based on the concept of Enhanced Recovery After Surgery (ERAS) for pelvic fractures.Methods:A prospective randomized control trial was conducted to include 114 emergency patients who had been admitted to Department of Orthopaedic Trauma, Beijing Jishuitan Hospital for surgical treatment of pelvic fractures from June 2019 to December 2020. Of them, 57 were assigned into an intervention group according to a random digits table. They were 42 males and 15 females, aged from 18 to 77 years and subjected to management of pelvic fractures with tentative perioperative ERAS approaches which were adjusted at different stages. The other random 57 patients were assigned into a control group. They were 40 males and 17 females, aged from 17 to 70 years and subjected to management of pelvic fractures with conventional rehabilitation approaches which included postoperative in-hospital consultation and guidance by rehabilitation physicians. The 2 groups were compared in terms of Majeed pelvis scores and Barthel indexes at postoperative 2, 6, 12 and 24 weeks, and visual analogue scale (VAS) pain scores and SF36 scores at postoperative 12 and 24 weeks.Results:A total of 105 patients (55 in the intervention group and 50 in the control group) were completely followed up for 151 to 254 d (mean, 177 d). The 2 groups were comparable due to no significant difference between them in the preoperative general data ( P>0.05). The Majeed scores (44±13, 67±16, 86±14 and 98±7) and Barthel indexes (57±13, 79±16, 95±8 and 100±2) at postoperative 2, 6, 12 and 24 weeks in the intervention group were significantly higher than those in the control group [(35±16, 51±16, 73±14 and 91±12) and (45±19, 67±18, 86±12 and 98±4)] (all P<0.05). At postoperative 12 and 24 weeks, the SF-36 scores (129±15 and 141±6) in the intervention group were significantly higher than those in the control group (114±15 and 131±12) ( P<0.05). There was no significant difference in the pain degree between the 2 groups ( P>0.05). Conclusion:In management of pelvic fractures, compared with conventional perioperative rehabilitation approaches, the perioperative ERAS rehabilitation approaches may improve early functional outcomes and thus help the patients restore their activities of daily living earlier.
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OBJECTIVE：To investigate the effects of Guizhi ful ing capsules and its principal components （paeoniflorin， paeonol and amygdalin ）on the intestinal flora of primary dysmenorrhea model rats. METHODS ：Female SD rats were randomly divided into normal group ，model group ，capsule group（Guizhi fuling capsule ，1 000 mg/kg），paeoniflorin group （15.0 mg/kg）， paeonol group （10.3 mg/kg）and amygdalin group （12.1 mg/kg），with 6 rats in each group. Except for normal group ，other groups were given estradiol benzoate subcutaneously on the back of rats and oxytocin intraperitoneally to induce primary dysmenorrhea model. From the 4th day after subcutaneous injection of estradiol benzoate ，normal group was given constant volume of normal saline intragastrically ；model group was given constant volume of 0.5％CMC-Na solution intragastrically ；administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. The writhing times and the contents of and MDA in uterus tissue of rats were determined ，and then com the contents of short-chain fatty acids （SCFAs）such as acetic acid，propionic acid ，butyric acid in colonic contents were detected by GC method. Using diver sity index as index ， Rep-PCR and Eric-PCR were used to evaluate the d iversity of intestinal flora in feces of rats. RESULTS ：Compared with normal group，the writhing times of rats were increased significantly in model group ；the contents of NO and MDA in uterus were increased significantly ，while the contents of acetic acid ，propionic acid and butyric acid in colonic contents and total content of SCFAs were decreased significantly （P＜0.05 or P＜0.01）；the number of DNA electrophoresis bands of intestinal flora was significantly reduced ，the brightness of most bands was significantly reduced ，and the diversity indexes （by Rep-PCR and Eric-PCR method ，hereinafter）1 h after administration were significantly reduced （P＜0.05 or P＜0.01）. Compared with model group，writhing times of rats were decreased significantly in capsule group ，paeoniflorin group and paeonol group ；the contents of NO in uterus of rats in capsule group and paeoniflorin group as well as the contents of MDA in capsule group ，paeoniflorin group and paeonol group were decreased significantly （P＜0.05 or P＜0.01）；the propionic acid content and total content of SCFAs in colon of rats in capsule group ，the contents of acetic acid ，propionic acid and butyric acid ，total content of SCFAs in paeoniflorin group as well as the contents of propionic acid and butyric acid ，total content of SCFAs in paeonol group were increased significantly；the content of isovaleric acid was decreased significantly in paeoniflorin group （P＜0.05 or P＜0.01）；DNA electrophoresis bands and its brightness of intestinal flora changed to different extents in administration groups ，and the diversity indexes of intestinal flora 1 h after administration were increased significantly in capsule group and paeoniflorin group ，while those indexes were decreased significantly in paeonol group and amygdalin group （P＜0.05 or P＜0.01）. CONCLUSIONS ：Guizhi fuling capsules can significantly reduce writhing times and the contents of NO and MDA in uterus of primary dysmenorrhea model rats. At the same time ，the capsules also can regulate SCFAs content in colonic contents and intestinal flora diversity of rats. The above effects may be related to paeoniflorin and paeonol in the capsules.
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Previous studies have shown that spinosin was implicated in the modulation of sedation and hypnosis, while its effects on learning and memory deficits were rarely reported. The aim of this study is to investigate the effects of spinosin on the improvement of cognitive impairment in model mice with Alzheimer’s disease (AD) induced by Aβ1–42 and determine the underlying mechanism. Spontaneous locomotion assessment and Morris water maze test were performed to investigate the impact of spinosin on behavioral activities, and the pathological changes were assayed by biochemical analyses and histological assay. After 7 days of intracerebroventricular (ICV) administration of spinosin (100 µg/kg/day), the cognitive impairment of mice induced by Aβ1–42 was significantly attenuated. Moreover, spinosin treatment effectively decreased the level of malondialdehyde (MDA) and Aβ1–42 accumulation in hippocampus. Aβ1–42 induced alterations in the expression of brain derived neurotrophic factor (BDNF) and B-cell lymphoma-2 (Bcl-2), as well as inflammatory response in brain were also reversed by spinosin treatment. These results indicated that the ameliorating effect of spinosin on cognitive impairment might be mediated through the regulation of oxidative stress, inflammatory process, apoptotic program and neurotrophic factor expression, suggesting that spinosin might be beneficial to treat learning and memory deficits in patients with AD via multi-targets.
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Animals , Humans , Mice , Alzheimer Disease , B-Lymphocytes , Brain , Brain-Derived Neurotrophic Factor , Cognition Disorders , Hippocampus , Hypnosis , Learning , Locomotion , Malondialdehyde , Memory Disorders , Memory , Neuroprotection , Neuroprotective Agents , Oxidative Stress , WaterABSTRACT
OBJECTIVE:To optimize the flash extraction technology for Guizhi fuling capsule. METHODS:Using blade speed,liquid-solid ratio,extraction time as investigation factors,using the comprehensive evaluation values consisting of transfer rates of gallic acid,paeoniflorin,benzoic acid,cinnamic acid,benzoyl paeoniflorin,amygdalin as investigation index,single fac-tor test was combined with Box-Behnken design-response surface method to optimize the flash extraction technology for Guizhi ful-ing capsule. Verification test was conducted and compared with conventional decoction extraction method(extracting twice,totally 240 min). RESULTS:The optimal flash extraction technology for Guizhi fuling capsule was using water as extraction solvent with blade speed of 5600 r/min and liquid-solid ratio of 10.5:1,extracting for 60 s. The average comprehensive evaluation value of veri-fication test was 85.40%(n=3),with relative error of 0.15% to the predicted value(85.55%),higher than decoction extraction method(72.65%). CONCLUSIONS:Optimized flash extraction technology for Guizhi fuling capsule is efficient and rapid.
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OBJECTIVE:To optimize the dry granulation technology conditions for Yinqiao baidu tablet. METHODS:Using granulating difficulty degree and disintegration time as investigation indexes,ratio and amount of accessories microcrystalline cellu-lose and compressible starch in Yinqiao baidu tablet,moisture content of the sprayed powder were screened. Using yield of particle and angle of repose as indexes,L9(34)orthogonal test was used to optimize the wheel pressure,rotating speed and feeding speed in dry granulation technology,and verification test was conducted. RESULTS:The ratio of microcrystalline cellulose and compress-ible starch was 7:3,and mixing ratio of the two with spray powder+inclusion compound was 1:5. The moisture content of spray powder was controlled in 1％-2％. The optimal technology was as follow as wheel pressure of 3.5 MPa,roller speed of 4 r/min and feeding speed of 10 r/min. In verification test,average yield of particle was 69.2％ and angle of repose was 31.5 °. Transfer rate of chlorogenic acid had reached over 92％,and RSD of each index was below 2.53％(n=3). CONCLUSIONS:Each index of parti-cle prepared by optimized accessories formulation and technology shows good reproducibility and feasibility,and the technology is stable and suitable for production.
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OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.
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Box-Behnken design-response surface methodology was used to optimize the transformation from chlorogenic acid to neochlorogenic acid.Based on single factor experiments,the experiment design were developed by box-benhnhen central composite design with causal factors of the reaction temperature,time and PH to neochlorogenic acid.The optimum transformation conditions were as follow:reaction temperature at 107℃,reaction time of 60 min,the PH of 4.72.Under the optimum extraction technology conditions,the productivity of neochlorogenic acid was 64.20％.Neochlorogenic acid was isolated and purified.The determination and characterization of neochlorogenic acid was detected by HPLC,1H-NMR,13C-NMR and ESI-MS.The results showed that the content of neochlorogenic acid reached to 98.78％ and the yield of 87.37％.
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OBJECTIVE:To optimize the ultrasonic extraction technology for Tianshu capsules. METHODS:Using the transfer rate of active ingredient ferulic acid in Ligusticum chuanxiong and gastrodin in Gastrodia elata of Tianshu capsules as investigation indexes,L9(34)orthogonal design test was adopted to investigate the effects of ethanol volume fraction,ethanol amount,extraction time and ultrasonic power on the extraction rate. Ultrasonic extraction technology for Tianshu capsules was optimized,and verifica-tion test was carried out. RESULTS:The optimized extraction technology for Tianshu capsules was as follow as 8-fold 70% etha-nol,extracting twice under 350 W,extracting 1.5 h every time. Results of verification test showed the average transfer rate was 94.06% for ferulic acid(RSD=0.18%,n=3)and 95.02% for gastrodin(0.47%,n=3). CONCLUSIONS:The optimized tech-nology is rapid,simple,stable and feasible,and can be used for extracting the active ingredients in Tianshu capsules.
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OBJECTIVE:To establish the quality standard for the leaves of Nauclea officinalis. METHODS:Microscopic identi-fication and TLC methods were used for qualitative identification of N. officinalis. The moisture and ash of medicinal materials were determined. HPLC method was adopted to determine the content of strictosamide in medicinal materials. The determination was per-formed on Lichrospher C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1 mL/min,injection volume was 10 μL. The detection wavelength was set at 226 nm,and the column temperature was 30 ℃. RESULTS:Microscopic identification of the leaves of N. officinalis had strong characteristics,and TLC sports were clear and well-separated without interference from negative control. The linear range of strictosamide were 0.0105-0.21 mg/mL(r=0.9995);RSDs of precision,stability and reproducibility tests were all lower than 2.0%. Average recoveries were 96.25-101.82%(RSD=1.86%,n=9). Total ash of medicinal materials was 5.27%-6.44%,acid insoluble ash was 0.23%-0.36%,water content was 9.48%-11.46%,strictosamide was 0.124%-1.003%. CONCLUSIONS:Established standard can be used for quality evaluation of N. officinalis.
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AIM To establish an HPLC method for the simultaneous content determination of four constituents in Qingzhiyi Tablets (Puerariae lobatae Radix,Phyllanthi Fructus,Salviae miltiorrhizae Radix et Rhizoma,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 30 ℃ thermostatic Kromasil C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of 0.1％ formic acid-methanol-acetonitrile flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 270 nm.RESULTS Gallic acid,puerarin,salvianolic acid B and tanshinone Ⅱ A showed good linear relationships within the ranges of 11.95-382.48,14.23-455.28,10.77-344.68 and 3.89-124.32 μg/mL,whose average recoveries were 99.96％,100.92％,98.87％ and 97.67％ with the RSDs of 1.09％,1.30％,1.11％ and 1.22％,respectively.CONCLUSION This sensitive,simple and accurate method can be used for the quality control of Qingzhiyi Tablets.
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Aim To investigate the protective effects of Diterpene Ginkgolides Meglumine Injection(DGMI)on SY5 Y cells damaged by oxygen-glucose deprivation and its functional mechanisms.Methods After 4 h of OGD,the cells were treated with 25 mg·L-1 drugs for 1 h.Subsequently,cell viabilities were measured by cell counting kit-8(CCK-8 kit)and cell apoptosis was measured by flow cytometric analysis.Furthermore, the mitochondrial membrane potential was detected by rhodamine123 staining.The levels of phospho-p38, phospho-p53,Bcl-2,Bax and cleaved caspase-9/3 were evaluated by western blot.Results DGMI signif-icantly increased the cell viabilities of SY5 Y cells dam-aged by OGD,and reduced OGD-elicited dissipation of mitochondrial membrane potential and cell apoptosis. Furthermore,DGMI also reduced p-p38,p-p53,Bax/Bcl-2 ratio,cleaved caspase-9 and cleaved caspase-3. Conclusion DGMI shows good neuroprotective effects on SY5 Y cells after oxygen-glucose deprivation.The underlying mechanisms may be associated with the sup-pression of p38/p53/Bcl-2 /caspase-9/caspase-3 sig-naling pathway.
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Aim To investigate the effects of diterpene ginkgolides meglumine injection (DGMI ) on amino acids and monoamine neurotransmitters in rats with cerebral ischemia/reperfusion injury.Methods In-traluminal suture was applied to establish middle cere-bral artery occlusion (MCAO/R)model with ischemia for 1.5 h and reperfusion for 24 h.After the adminis-tration of DGMI (i.v.),the levels of amino acid and monoamine neurotransmitters in brain tissue were de-tected through HPLC-ECD.Results DGMI down-reg-ulated the concentrations of aspartic acid, glutamic acid,glycine and γ-aminobutyric acid which were in-creased in MCAO/R group.DGMI also reduced the levels of norepinephrine epinephrine,glyoxylic acid, serotonin and 5-HIAA in cortex and hippocampus,and increased adrenaline content compared to the model group.Conclusion DGMI exhibits a protective role in rats with cerebral ischemia /reperfusion injury through regulating amino acids and monoamine neuro-transmitters.
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OBJECTIVE:To establish a method for the simultaneous determination of 5 unsaturated fatty acids in Perilla oil cap-sule. METHODS:With the reference material of α-linolenic acid methyl ester,GC was used to determine and calculate the relative correction factors of α-linolenic acid methyl ester with methyl palmitate,methyl stearate,methyl oleate and linoleic acid methyl es-ter,and the correction factors were used to calculate the contents of 5 unsaturated fatty acids;the column was Agilent Innowax cap-illary column,the detector was FID,the inlet temperature was 230 ℃,the detector temperature was 250 ℃,the gas flow rate was 20 ml/min(nitrogen),40 ml/min(hydrogen)and 350 ml/min(air),split ratio was 30 to 1,the column temperature was 190 ℃, and injection volume was 1 μl. RESULTS:The linear range was 0.018-0.792 μg(r=0.9994)for methyl palmitate,0.0016-0.0176μg(r=0.9993)for methyl stearate,0.0056-0.2464 μg(r=0.9999)for methyl oleate,0.003-0.132 μg(r=0.9990)for linoleic acid methyl ester and 0.018-0.792 μg(r=0.9998) for α-linolenic acid methyl ester;RSDs of precision,stability and reproducibility tests were lower than 5%;recoveries were 98.990%-101.70%(RSD=0.720%,n=6) for methyl palmitate,99.599%-100.699%(RSD=0.368%,n=6) for methyl stearate,98.996%-101.680%(RSD=1.240%,n=6) for methyl oleate,99.813%-100.963%(RSD=0.434%,n=6)for linoleic acid methyl ester and 97.185%-99.602%(RSD=0.874%,n=6)for α-linolenic acid methyl es-ter. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of methyl palmitate,methyl stearate,methyl oleate,linoleic acid methyl ester,α-linolenic acid methyl ester in Perilla oil cap-sule.
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Aim To investigate the effects of Sanjie Zhentong capsule on cultured mouse myometrial cell contraction induced by prostaglandin F2α( PGF2α) , and to elucidate the mechanism of Sanjie Zhentong capsule in treating dysmenorrheal. Methods Primary mouse myometrial cells were cultured and identified. Intracel-lular calcium ( [ Ca2+] i ) was monitored under a Flex-Station 3 Benchtop Multi-Modo Microplate Reader u-sing Calcium 6-QF. Myometrial cells were labeled to observe the changes of contraction. The expressions of calmodulin ( CaM ) , myosin light chain kinase ( ML-CK) , myosin light chain phosphorylation( p-MLC20 ) in mouse uterine smooth muscle cells ( USMCs ) were determined by immunofluorescence. Results Sanjie Zhentong capsule suppressed the intracellular[Ca2 + ]i inflow and reduced the areas of myometrial cells induced by PGF2α. Then it significantly decreased CaM and p-MLC20 levels. Conclusion Our results indicate that Sanjie Zhentong capsule has inhibitory effect on dysmenorrheal induced by PGF2α. Furthermore,its major mechanism may be related with the regulation of intracellular[ Ca2 + ]i inflow and the levels of CaM and p- MLC20.
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This study aimed at investigating the antiviral constituents from the active fractions of Tong-An (TA) injection.In this study,the active constituents of TA injection were screened by LPS-induced PGE2 production mode to detect the contents of PGE2.The chemical constituents were isolated by HP-20 macroporous resin,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography and preparative and semi-preparative HPLC.The structures were identified by spectral data and physicochemical property.As a result,the 95％ ethanol eluate of TA injection on the macroporous adsorption resin column was proved to be the active fraction of TA injection.Seventeen compounds were isolated from TA injection and identified as syringaresinol (1),N-Trans-Feruloyltyramine (2),chelerythrine (3),sinomenine (4),coptisine (5),sanguinarine (6),chelidoniny (7),magnoflorine (8),allocryptopine (9),protopine (10),farrerol (11),dihydrosanguinarine (12),heptadec-(9Z)-enoic acid (13),chlorogenic acid (14),cryptochlorogenin acid (15),3,5-di-O-caffeoylquinic acid (16) and 4,5-di-O-caffeoylquinic acid (17).PGE2 inhibitory activities of these compounds were determined,among which six compounds presented inhibitory activities against PGE2.It was concluded that all the isolated compounds from TA injection were firstly reported with the favorable inhibitory activities of compounds 2,5,9,10,11,12 against PGE2.
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OBJECTIVE:To establish a method for the determination of 7 residual solvents(ethanol,n-hexane,benzene,tolu-ene,xylene,styrene,divinylbenzene)in Liuwei dihuang glycoside. METHODS:The column was DB-624 capillary column,carri-er gas was nitrogen,flow rate was 5.0 ml/min;detector was a hydrogen flame ionization detector with temperature of 250 ℃(pro-grammed temperature);equilibrium temperature was 80 ℃,sample loop temperature was 90 ℃,and transfer line temperature was 100 ℃;the equilibrium time of vial heating was 30 min,sample loop filling time was 0.05 min,injection time was 1.0 min;the carrier gas pressure was 95 kpa,and the vial pressure was 60 kpa. RESULTS:The linear range was 25-500 μg/ml for ethanol(r=0.998 7),0.025-10μg/ml for n-hexane(r=0.998 8),0.025-10μg/ml for benzene(r=0.999 9),0.1-40μg/ml for toluene(r=1.000 0),0.25-100 μg/ml for xylene(r=0.999 9),0.5-500 μg/ml for styrene(r=1.000 0) and 0.5-500 μg/ml for divinylbenzene (r=1.000 0);RSDs of precision,stability and reproducibility tests were lower than 4%;recoveries were 99.60%-102.70%(RSD=1.08%,n=9),90.70%-100.30%(RSD=4.51%,n=9),100.10%-109.80%(RSD=3.82%,n=9),99.50%-110.00%(RSD=4.40%,n=9),100.00%-109.10%(RSD=3.50%,n=9),93.40%-102.30%(RSD=3.73%,n=9) and 99.70%-101.70%(RSD=0.79%,n=9),respectively;the low limits of detection were 1.000,0.025,0.025,0.025,0.100,0.025,0.250 μg/ml respectively. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the determination of residual solvents(etha-nol,n-hexane,benzene,toluene,xylene,styrene,divinylbenzene)in Liuwei dihuang glycoside.