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In recent years, evidence-based medicine has developed rapidly, and its ideas and methods have been increasingly used in medical education. We have promoted the model, theory and practical methods of evidence-based medicine in the teaching of endocrinology standardized residency training, which has improved students' learning enthusiasm and initiative, and also enhanced students' mastery of professional knowledge and cultivation and innovation of clinical thinking. In view of the existing problems, this essay puts forward some solutions to help residents to meet the requirements of rapid developments in endocrinology and generally promote the standardization of residents' medical practice behavior.
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OBJECTIVE： To provide reference for the establishment of quality standard of Shenhuang liniment. METHODS： Qualitative identification of Sophora flavescens， Phellodendron chinense， Rhei Radix Et Rhizoma and Scutellaria baicalensis in Shenhuang liniment were carried out by TLC according to the method of 2015 edition of Chinese Pharmacopeia （part Ⅳ）. HPLC method was used to determine the contents of matrine and oxymatrine in Shenhuang liniment [Phenomenex Luna NH2 column， mobile phase consisting of acetonitrile-absolute ethanol-3％ phosphoric acid aqueous solution （80 ∶ 10 ∶ 10，V/V/V）， column temperature of 45 ℃， flow rate of 1.0 mL/min， detection wavelength of 220 nm， sample size of 5 μL]. RESULTS： Results of TLC showed that the corresponding spots of the same color were found in the corresponding positions of the chromatograms for test samples and reference substance/substance control； and the spots were clear， retardation factor was moderate， the separation degree was high， and the negative control had no interference. Results of HPLC showed that the linear range of matrine and oxymatrine were 203.60-1 221.60 ng（r＝0.999 4）and 210.08-840.30 ng（r＝0.999 7）， respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.0％ （n＝6）. Average recovery rates were 96.03％ and 100.93％， and RSDs were 2.55％ and 2.69％（n＝9）. CONCLUSIONS： Established method is specific， accurate and reproducible， and can be used for quality control of Shenhuang liniment.
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Objective To investigate the effect of fenofibrate on glucolipid metabolism and insulin sensitivity in lipoprotein lipase heterozygous knockout ( LPL+/-) mice, and to explore its mechanism. Methods LPL+/- mice and wild type ( WT) C57 mice were selected and divided into 3 groups ( n=6 each group):LPL+/-( FB) group, LPL+/-(W)group,andWTgroup.MiceinLPL+/-(FB)groupweregavagedwithfenofibrate(50mg·kg-1·d-1)for8 weeks. Mice in LPL+/-( W) and WT groups were orally fed with the same volume water as that in LPL+/-( FB) group for 8 weeks. Body weight was observed. Plasma triglyceride ( TG ) and free fatty acid ( FFA ) were measured. Intraperitoneal glucose tolerance test in 3 groups of mice were performed. The glucose area under the curve ( AUCG) and homeostasis model assessment for insulin resistance index ( HOMA-IR) were calculated. Insulin-stimulated Ser473 Akt phosphorylation in liver and skeletal muscle was measured by Western blot. Reactive oxygen species ( ROS) levels in liver and skeletal muscle were determined by dihydroethidium staining method and superoxide dismutase ( SOD) and catalase ( CAT) mRNA expression levels were detected by real-time PCR. Results Compared with LPL+/-( W) mice, body weight of LPL+/-( FB) mice was lowered, plasma TG and FFA levels were decreased by about 46.0％and 76.5％respectively, and fasting insulin level and HOMA-IR were decreased while there were no significant differences in fasting glucose level and AUCG between two groups. Insulin-stimulated Ser473 Akt phosphorylation levels in liver and skeletal muscle of LPL+/-mice were enhanced by fenofibrate. ROS level in skeletal muscle of LPL+/-( FB) mice was lower than that in LPL+/-( W) mice while there was no significant difference in ROS of liver between two groups. Fenofibrate significantly increased SOD and CAT mRNA expressions in skeletal muscle of LPL+/-mice, but not in liver. Conclusion Fenofibrate reduces body weight, ameliorates lipid metabolism, and improves insulin sensitivity in LPL+/- mice, with reduced oxidative stress.
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OBJECTIVE:To establish UPLC fingerprint of Gehua formula granules. METHODS:UPLC method were adopted. The determination was performed on Zorbax Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-water at the flow rate of 0.5 mL/min. The detection wavelength was set at 264 nm,column temperature was 25 ℃,and the sample size was 1 μL. Using tectorigenin as reference substance,UPLC chromatograms of 10 batches of Gehua formula granules were determined. The common peak identification and similarity evaluation were conducted by TCM Chromatogram Fingerprint Similarity Evaluation Sys-tem(2004 A edition). RESULTS:14 common peaks were identified in UPLC chromatograms of 10 batches of Gehua formula gran-ules and similarities were all higher than 0.90. UPLC chromatograms of 10 batches of samples were in good agreement with control fingerprint. CONCLUSIONS:Established UPLC fingerprint can provide reference for identification and quality evaluation of Gehua formula granules.
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AIM To establish the HPLC fingerprints of Cajanus cajan (L.) Millsp.leaves and to determine the contents of orientoside and luteolin.METHODS The analysis of 65％ methanol extract from C.cajan leaves was performed on a 25 ℃ thermostatic Agilent Zorbax SB-C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-1％ acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS There were twenty-one common peaks in ten batches of samples (S1-S10),whose similarities were more than 0.950,except for that of S3 (0.516).Orientoside and luteolin showed good linear relationships within the ranges of 0.089 5-3.960 μg and 0.015 5-0.408 μg,whose average recoveries were 99.43％ (RSD =1.32％) and 98.50％ (RSD =0.82％),respectively.The contents of two constituents in the samples from three growing areas (Guangdong,Yunnan and Hainan) showed obvious differences.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of C.cajan leaves.
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Objective To measure the fasting plasma ghrelin and obestatin concentrations in essential hyper-tension (EH)patients with obesity,and to observe their relationship with blood pressure,obesity index,insulin resist-ance (IR),blood glucose and blood lipid.Methods 68 hypertensive obese patients,60 normotensive obese patients and 65 healthy controls were included in the study.The fasting plasma obestatin and ghrelin were measured using a radioimmunoassay method.The ghrelin/obestatin ratio was calculated.Results Hypertensive obese patients had lower plasma ghrelin and obestatin compared with normotensive obese patients (t =3.771,P <0.01;t =4.373,P <0.01) and controls (t =16.451,P <0.01;t =17.862,P <0.01).Normotensive obese patients had lower plasma ghrelin and obestatin compared with controls (t =13.121,P <0.01;t =13.686,P <0.01 ).Hypertensive obese patients had higher ghrelin/obestatin ratio than controls (t =7.720,P <0.01).The ghrelin/obestatin ratio in normotensive obese patients was higher than controls (t =4.587,P <0.01).In hypertensive obese group,the plasma ghrelin and obestatin were negatively associated with BMI (r =-0.882,P <0.01;r =-0.806,P <0.01),SBP (r =-0.787,P <0.01;r =-0.837,P <0.01),DBP (r =-0.769,P <0.01;r =-0.810,P <0.01),and the HOMA-IR (r =-0.800, P <0.01;r =-0.810,P <0.01).In normotensive obese group,ghrelin and obestatin were all negatively associated with BMI (r =-0.577,P <0.01;r =-0.372,P <0.01)and HOMA-IR (r =-0.866,P <0.01;r =-0.662,P <0.01).The ghrelin/obestatin ratio was positively associated with BMI (r =0.460,P <0.01)and HOMA-IR (r =0.420,P <0.01).Conclusion The peripheral blood ghrelin,obestatin and ghrelin/obestatin ratio were significantly correlated with EH,obesity and IR.No correlation was observed between ghrelin/obestatin system and blood lipid or blood glucose.
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Objective To establish the quality standard for mongolian medicine Rhaponticum uniflorum (L.) DC. with modern analysis methods. Methods Chlorogenic acid and apigenin were identified by thin -layer chromatography (TLC). The chlorogenic acid content was determined by high performance liquid chromatograghy ( HPLC). And HPLC was performed on Kromat Universil C18 ( 4.6 mm × 250 mm, 5 μm) with methanol-water-acetic acid ( v/v/v, 15:85:0.85) as mobile phase, the detection wavelength was 326 nm, the flow rate was 1.0 mL/min, and the column temperature was 20℃. Results The samples and the reference substance shared the same color spots at the same sites. The linear range of chlorogenic acid was 0.044 66-0.446 6μg (r=0.999 8), and the average recovery rate was 99.84%. Conclusion The method is simple, accurate, and with good reproducibility, and can be used for the quality control of Rhaponticum uniflorum ( L. ) DC.
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Objective To investigate the glucolipid metabolism in lipoprotein lipase (LPL) gene knockout mice, and to explore the possible mechanisms of insulin resistance. Methods 16- and 40-week old LPL gene knockout heterozygous mice( LPL + / -) and wild type ( WT) C57 mice were selected and divided into 4 groups:16-week LPL+ / -(n=6), 16-week WT(n = 6), 40-week LPL+ / -(n = 6), and 40-week WT(n = 6) group. LPL activity of post-heparin serum was examined. Serum triglyceride( TG) and free fatty acid( FFA) were measuzed. Intraperitoneal glucose tolerance test(IPGTT) in 4 groups of mice were performed. The glucose area under the curve (AUCG) and homeostasis model assessment for insulin resistance index and β-cell function index ( HOMA-IR, HOMA-β) were calculated to evaluate insulin sensitivity and the function of islet β-cells. Serum malondialdehyde (MDA) and total antioxidant capacity ( TAOC) levels were determined by means of colorimetric method. Using dihydroethidium( DHE) fluorescent staining method, reactive oxygen species ( ROS) levels in liver and skeletal muscle were determined. Results LPL activity levels of both 16- and 40-week LPL+ / - mice were significantly lower than that in WT mice of the same age. Serum TG and FFA of 40-week old LPL+ / - mice were significantly higher than those in WT mice of the same age(P<0. 05), and they were also higher than those of 16-week old LPL+ / - mice(P<0. 05). IPGTT showed that compared with WT mice, blood glucose level in LPL+ / - mice was significantly higher than that in WT group at 30 and 120 minute(P<0. 05), and fasting insulin and HOMA-IR were increased significantly(P<0. 05). Serum MDA of 40-week old LPL+ / - mice was evidently higher than that in WT mice by the same week(P<0. 05), while TAOC level was lower than that of WT mice (P<0. 05). ROS in skeletal muscle of 16-week old LPL+ /- mice was significantly increased. Meanwhile, ROS in both liver and skeletal muscle of 40-week old LPL+ / - mice was significantly higher than that in WT mice of the same age. Conclusion As time goes by, lipid and glucose disorders of LPL+ / - mice are aggravating, and insulin resistance develops evidently. Insulin resistance in LPL+ / -mice with dyslipidemia may be related to oxidative stress.
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Objective To compare the differences in responsive ability and oxidative stress damage between type 2 diabetic rats and normal rats under acute sepsis associated with stress hyperglycemia.Methods GK rats with type 2 diabetes and Wistar rats were established critical ill models of sepsis by intraperitoneal injection of 5 mg/kg lipopolysaccharide(LPS).Before and after LPS injection,serum levels of proinflammatory cytokines tumor necrosis factor α (TNF-α),interleukin (IL)-1β3,and IL-6,and anti-inflammatory cytokine IL-10 were determined.Malondialdehyde (MDA) and total antioxidative capacity (T-AOC) levels in serum and tissues (lung,liver,kidney,heart) were measured by colorimetric determination.Results Before LPS injection,serum levels of TNF-α,IL-6,and IL-10 in GK rats were higher than those of Wistar rats.The serum levels of TNF-α,IL-6,and IL-10 after LPS injection were raised in both GK rats and Wistar rats (all P＜0.05).At baseline,serum MDA level of GK rats was higher than that of Wistar group [(25.76 vs 12.71) μmol/L,P＜0.05],while T-AOC level were lower [(0.60 vs 2.8) U/ml,P＜0.05].One hour after LPS injection,the serum MDA level in two kinds of rats significantly increased compared with those of their baseline levels [(32.30 and 20.19) μmol/L,both P＜0.05],while their serum T-AOC level decreased [(0.20 and 2.08) U/ml,P＜0.05]).There were no signigicant differences in the change trend of serum MDA and T-AOC between the two kinds of rats.MDA and T-AOC levels in tissues had no significant difference before and after LPS injection in GK and Wistar rats (P＞0.05).Conclusions The rats with type 2 diabetes had lowered level of proinflammatory factors and raised level of anti-inflammatory factors under acute sepsis,presenting better tolerance and lowered probability of inflammatory diffusion compared with normal rats.