BACKGROUND: Non-enzymatic glycation of proteins is involved in the complications of diabetes mellitus. Previous experiments have demonstrated that total flavones of buckwheat flower (TFBF) could improve carbohydrate tolerance. However, it is little known whether TFBF inhibit the non-enzymatic glycation of proteins.OBJECTIVE: To investigate the influences of TFBF on the non-enzymatic advanced glycation end products (AGEs) of proteins in vivo and in vitro.DESIGN: Completely randomized controlled trial.SETTING: Department of Pharmacology, North China Coal Medical College.MATERIALS: Totally 75 adult SD rats , of clean grade, weighing (200±20) g, including 38 female rats and 37 male rats, were provided by the institute of experimental animals, Chinese Academy of Medical Sciences (Certification No. SCXK11-00-0006). TFBF was extracted by our laboratory from flowers of buckwheat. The blood glucose kit was purchased from Beijing Biosino Biotechnology Company Ltd. Penicillin (Batch No.031020, 8×105 U) and streptomycin (Batch No. 030920, 1×106 U) were purchased from North China Pharmaceutical Company. Streptozotocin and BSA were purchased from Sigma Company. Fructosamine kit was purchased from Nanjing Jiancheng Bioengineering Institute, and the other chemicals were analytical pure produced domestically.METHODS: This experiment was carried out in the Department of Pharmacology, North China Coal Medical College from March to October 2004.In the first experiment, in vivo macromolecular AGEs was measured: ①Modeling and grouping: Rats were divided into 3 groups according to body mass: Normal control group (n=15), the rats were treated with 8 mL/kg normal saline intraperitoneally. Streptozotocin-treated group (n=45), the rats were fasted for 16 hours and then treated with 80 mg/kg streptozotocin of 8 mL/kg intraperitoneally. Twenty-two hours later, the blood of all rats was harvested from vena caudalis to measure the level of blood sugar.Those with fasting blood glucose ≥ 15 mmol/L were acted as diabetic rats.Streptozotocin-treated group were divided into 3 subgroups, 15 rats in each subgroups. Each rat was given intragastric administration of 0.1, 0.2 and 0.4 g/kg TFBF. Model group (n=1S): Rats were only treated with 80 mg/kg streptozotocin of 8 mL/kg . The rats in normal control group and model group were given the same volume of salt water. The administration was once a day for 12 weeks successively. ②Measurement of fasting blood glucose: After the last administration, the rats of streptozotocin-treated group were fasted for 12 hours and the blood was harvested from vena caudalis. The fasting blood glucose was measured by glucose oxidase method. ③The levels of blood plasma and nephridial tissue fructosamine and macromolecular AGEs were measured: The rats of each group were anesthetized with ethyl ether on the second day following the last administration. Blood was chosen from carotid artery, and plasma was separated.Kidneys were taken at the same time, prepared into 100 g/L tissue homogenate and centrifuged at low temperature. The levels of fructosamine in plasma and the supernatant fluid of kidney homogenate were measured according to the instructions of the kit. AGEs in plasma and renal tissue were determined by fluorospectrophotometer. The products of macromolecular AGEs were calculated. In the second experiment, in vitro macromolecular AGEs were measured as below: 0.01, 0.05, 0.10 mg/L TFBF of 6 mLrespectively was prepared with solution A (0.2 mol/L glucose, 2×l06 U/Lpenicillin, 2×106 U/L streptomycin , PBS containing 20 g/L bovine serum albumin). Control groups were set: ① without TFBF, ② without TFBF and glucose, ③ without BSA, ④ without glucose. Five parallels of each sample were sterilized by filtration and incubated in the attemperator at 37 ℃. The fluorescence of AGEs (F) in the culture was determined at the 4th, 8th and 12th weeks. Inhibition ratio (IR) was calculated and the inhibition of TFBF on AGEs was observed.MAIN OUTCOME MEASURES: In the first experiment, the levels of fasting blood glucose, fructosamine in kidney and plasma, and AGEs were measured. In the second experiment, the inhibition of TFBF on AGEs in vitro was measured.RESULTS: In the first experiment, 75 rats were involved, and 56 successful rats entered the stage of result analysis. The levels of blood glucose,fructosamine in kidney and plasma of rats in the model group were significantly higher than those of rats in the normal control group (t=7.572,10.186, 5.794,P ＜ 0.01 ). The level of blood glucose of rats in the 3 subgroups was significantly lower than that of rats in the model group (t=3.357,4.382,3.938,P ＜ 0.05-0.01); The levels of fructosamine in kidney and plasma of rats in the 0.2 and 0.4 g/kg TFBF groups were significantly lower than those in the model group (t=5.109, 4.605, 3.731,3.097,P ＜ 0.05-0.01 ). The levels of AGEs in plasma and kidney of rats in the model group were significantly higher than those in the normal control group (t=6.463, 12.704,P ＜ 0.01 ), while the levels of AGEs in plasma of rats in the streptozotocin-treated group were similar to those in the model control group (P ＞. 0.05), and those in kidney of rats in the streptozotocintreated subgroups were significantly lower than those in the model group (t=9.845, 12.799, 12.899,P ＜ 0.01 ). In the second experiment, the level of macromolecular AGEs of each group was gradually increased with ime.TFBFcould inhibit the formation of macromolecular AGEs in dose- and time-dependent manner.CONCLUSION: TFBF obviously inhibited the formation of AGEs of proteins in vivo and in vitro.

Aim To explore the vasodilative effect on rat thoracic aortic ring of total flavonoids of Buckwheat flowers and leaves(TFBFL) and the underlying mechanisms.Methods Isometric tension measurements were used to study the effect of TFBFL on isolated rat thoracic aorta rings.Laser scanning confocal microscope was employed to measure the concentration of intracellular free calciums.Results In aorta rings precontracted with phenylephrine or potassium chloride,TFBFL caused a dose-dependent relaxation in both endothelium-intact and denuded rings and the relaxant effect of TFBFL was more potent on endothelium-intact aorta rings than that on endotheliumdenuded aorta rings(P

Objective:To observe the effect of Radix Paeoniae Bubra(RPB)on NADPH oxidase p22phox,monocyte chemotactic protein-1(MCP-1)mRNA expression and neointimal hyperplasia after carotid artery balloon injury in cholesterol-fed rabbits.Methods:The rabbit model of carotid balloon injury was established adopting Clowes method,and treated with extract of RPB.Component of neointima and expression of proliferating cell nuclear antigen(PCNA)and macrophage was determined by immunochemical stain.The collagen of typeⅠwas detected by special staining for blood vessels and the area of neointima was measured by image assay system.Expression of NADPH oxidase p22phox mRNA,MCP-1 mRNA was detected by in situ hybridization and transcription-polymerase chain reaction.Results:RPB can attenuate the neointima area and proliferation of collagen typeⅠinduced by balloon-injury,remarkable prevent the formation of restenosi,and down-regulate expression of NADPH oxidase p22 and MCP-1mRNA significantly and decrease the degree of macrophages infiltration especially in vessel wall of injuring carotid artery.Conclusions:RPB inhibited NADPH oxidase P22Phox and MCP-1 mRNA expression,and modestly reduced neointimal hyperplasia,which might be partly attributed to its antioxidant and inflammatory effects.

Aim To investigate the influence of total fl avones of buckwheat flower (TFBF) on the productivity of the non-enzymatic advanced glycation end products (AGEs) in vivoand vitro. Methods TFBF in different dosages (0.1 g?kg -1?d -1,0.2 g?kg -1?d -1,0.4 g?kg -1?d -1) was taken orally by streptozotocin-induced diabetic rats for 12 wk. After the treatment, blood glucose, fructosamine and AGEs in plasma and kidney were measured. Meanwhile, glucose and bovine serum albumin (BSA) were incubated with TFBF at different concentrations (0.01 mg?L -1,0.05 mg?L -1,0.10 mg?L -1) respectively for 4,8,12 wk.The fluorescence intensity of glycated BSA was detected by a spectrophotometer BSA was detected spectrophotometer.Results TFBF significantly lowered the level of blood glucose in diabetic rats (P

Objective:To observe the inhibitory effect on ?-glucosidase of 10 kinds Chinese herbs and to screen the Chinese herbal medicines which have great inhibitory effect on ?-glycosidase.Methods:The ?-glucosidase was extracted from small intestine of rat.The amount of glucose was measured with produced from substrate of malt sugar.The inhibitory effect of 10 kinds of Chinese herbs on ?-glucosidase was observed by this enzyme reaction system.Then disposable gastric perfused malt sugar(2 g/kg) and the extraction screened at the same time,detected the levels of blood glucose after 60 min.The positive control group is acarbose(ACAR) group.Results:The three kinds Chinese herbs(Chishao,Shanzhuyu and Sangbaipi) showed very good inhibitory activities,and they showed obviously concentration-effect curve relationship.Among them,the inhibitary activity of Sangbaipi is stronger than Chishao and Shanzhuyu.While the dose of Sangbaipi reached 10mg/mL,the inhibition rate arrived 80%,which effect was equivalet to the dose of 1mg/ml acarbose.The results of postprandial blood glucose in vitro showed us:Sangbaipi,Chishao and Shanzhuyu can inhibit postprandial blood glucose levels in rats that have been disposable gastric perfused malt sugar after 60min(P0.05).Conclusion:The three kinds of Chinese herbs(Chishao,Shanzhuyu and Sangbaipi) can inhibit ?-glucosidase activity obviously in both vitro and vivo.

Objective To compare the difference in recurrence rates between the endovenous laser therapy(EVLT) combined with percutaneous continuous circumsature (PCCS) and simplex EVLT following the treatment of great saphanous vein incompetence through the introduction of propensity score matching (PSM).Methods T the baseline data of 170 patients diagnosed with great saphanous vein incompetence who were treated in Punan Hospital in Pudong New District of Shanghai from 2009 to 2014 were retrospectively analyzed,of which underwent EVLT were 87 cases as EVLT group and EVLT combined with PCCS were 83 cases as EVLT +PCCS group.The groups covariate were balanced based on the PSM function of SPSS software using 1 ∶ 1 nearest neighbor matching method.The recurrence rates of the two groups were estimated by Kaplan-Meier method and the differences between the two groups were evaluated by Log-rank test.Results Sixty-seven pairs of patients were successfully matched.No significant difference between the two matched groups in the basic clinical features.Before PSM,the 1,2,and 3 year cummulative recurrence rates were 3.5％,5.4％ and 7.3％ in the EVLT group,and 0.9％,4.7％ and 4.7％ in the EVLT+PCCS group,respectively,there were no statistically significant differences between the two groups by Log-rank test (P =0.491).After PSM,the 1,2,and 3 year cummulative recurrence rates were 5.2％,5.2％ and 7.1％ in the EVLT group,and 0％,1.0％ and 1.0％ in the EVLT+PCCS group,there were statistically significant differences between the two groups (P =0.031).Conclusion The PSM methods can effectively balanced the covariates of groups in non-randomised study.EVLT combined with PCCS can effectively reduce the recurrence rate after the treatment of great saphanous vein incompetence.

Objective To evaluate the value and safety of transbronchial needle aspiration (TBNA) in the diagnosis of lung cancer. Methods The cytologic diagnosis of TBNA in 82 patients with enlarged hilar and/or mediastinal lymphnodes or lesions adjacent to the bronchial wall were analyzed retrospectively. All specimens were detected by the ThinPrep cytologic test. Results There were 43 positive cases in the 82 patients, and the positive rate was 52.4 %. There were 18 SCLCs,11squamous cancers, 9 adenocarcinomas and 5 undefinable cancers, respectively. There were 39 patients with local bronchial wall swelling accompanied with abnormal mucosae. TBNA, douche, brushing and forcep biopsy were applied, and the diagnostic rate was 64.1%, 7.7 %, 25.6 % and 48.7 %, respectively. The total positive rate was 76.9 %. 43 patients with normal mucous membrane only underwent TBNA. 18 cases were positive, and the positive rate was 41.9 %. There was no obvious complication in the 82 patients. Conclusion The technique of TBNA enlarged the inspection scope of bronchoscopy. It has significant meaning to the diagnosis of lung cancer. TBNA was an useful and safe method in clinical application and could be used widely.

OBJECTIVETo explore the effect and mechanism of flavones of buckwheat flower and leaf (FBFL) on lowering blood glucose and improving insulin resistance in type 2 diabetic rats.

METHODSeventy healthy male Wistar rats were used in this trial. Ten of them were selected randomly as normal group; the others were given fat milk by intragastric administration daily, from the 14th day on, low dose tetraoxypyrimidine was added by intraperitoneal injection every other day for three times. Rats with fasting (72 hours after the last injection) blood sugar > or = 16.7 mmol x L(-1) and K(IPT) < 60% of normal group were selected as type 2 diabetic model with insulin resistance, which were randomly divided into 5 groups: model group. LGLT group; low, moderate and high dosage FBFL groups (L-FBFL; M-FBFL; H-FBFL). Every rat was treated accordingly for 4 weeks; then FBG, FFA, INS were detected and ISI was calculated to evaluate the degree of insulin resistance. Liver PTP1B expression was determined by immunohistochemistry method. staining were observed by light microscopy.

RESULTFBFL could dose-dependently inhibit the rising of FBG, FFA, INS, improve the state of insulin resistance and reduce the expression level of liver PTP1B.

CONCLUSIONFBFL could effectively improve insulin resistance in type 2 diabetic rats induced by tetraoxypyrimidine and fat milk and showed dose-dependence relationship.

Animals ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Fagopyrum ; chemistry ; Flavones ; administration & dosage ; Flowers ; chemistry ; Gene Expression ; drug effects ; Humans ; Insulin Resistance ; Liver ; drug effects ; metabolism ; Male ; Plant Extracts ; administration & dosage ; Plant Leaves ; chemistry ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo evaluate the prognosis of cervical intraepithelial neoplasia grade 1 (CIN1) at different follow-up time points in Chinese women and the relationship with high-risk human papillomavirus (HR-HPV) infection.

METHODSBiopsy-confirmed CIN1 women were followed up from cervical cancer screening cohorts established during 1999 to 2008 in Xiangyuan county, Yangcheng county, Qinxian county and Wuxiang county, Shanxi Province.In each follow-up visit, participants were examined by visual inspection with acetic acid, liquid-based cytology and HR-HPV DNA testing. Those with any positive results received colposcope and biopsies. The cumulative incidence rates of CIN grade 2 or worse (CIN2+) and CIN grade 3 or worse (CIN3+), regression rates and persistent rates were calculated using pathological findings as a gold standard. The risks of progression related with HR-HPV were evaluated stratified by baseline and follow-up HR-HPV status.

RESULTSA total of 228, 224, 261 and 105 CIN1 women received the 1-year, 2-year, 6-year and 11-year follow-up exams, respectively. The cumulative incidence rate of CIN2+ among baseline HR-HPV positive women was 4.8% (6/126), 10.7% (16/150), 16.9% (29/172) and 35% (19/55) in the above follow-up visits, respectively, and their risk of progression was 2.7(95%CI:0.3-22.0), 2.9 (95%CI:0.7-12.1), 12.0 (95%CI:1.7-86.2) and 30.6 (95%CI:1.9-493.5) times higher than baseline HR-HPV negative women. Moreover, the cumulative incidence of CIN2+ among women with positive HR-HPV both at baseline and follow-up visit was 11% (6/55), 14% (6/42), 17% (10/60) and 50% (13/26) in the above follow-up visits, respectively.No new CIN2+ cases were found among those with negative HR-HPV both at baseline and follow-up visits.

CONCLUSIONGiven that CIN1 progression is related to HR-HPV infection, different follow-up intervals and strategies for CIN1 should be taken according to HR-HPV infection status.

Aged ; Biopsy ; Cervical Intraepithelial Neoplasia ; Disease Progression ; Female ; Humans ; Papillomavirus Infections ; Prognosis ; Prospective Studies ; Uterine Cervical Neoplasms

【Objective】 To investigate the correlation of serum exosomal microRNA-301a (miR-301a) with cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. 【Methods】 A total of 211 patients with diabetic nephropathy treated with peritoneal dialysis from June 2019 to June 2020 in the First Hospital Affiliated of Hebei North University were selected as study subjects. Serum exosomal miR-301a was detected by real-time fluorescence quantitative polymerase chain reaction. The patients were divided into high miR-301a group and low miR-301a group based on the median of miR-301a; the clinical data of the two groups were compared. The correlation of miR-301a with high-sensitivity C-reactive protein (hs-CRP) was analyzed by Spearman. Linear regression was applied to analyze the factors associated with the effect of miR-301a. The patients were followed up for two years. Kaplan-Meier and Log-Rank were conducted to compare the cumulative incidence of cardiovascular and cerebrovascular events between the two groups, and COX regression and restricted cubic spline were used to analyze the level-effect relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis. 【Results】 Thirty-seven cases (17.54%) of cardiovascular and cerebrovascular events occurred during follow-up. The hs-CRP level and dialysis duration were lower in low miR-301a group than in high miR-301a group (P<0.05). There was a positive correlation between miR-301a and hs-CRP (rs=0.237, P=0.001). Linear regression analysis showed that hs-CRP was independently associated with miR-301a (P<0.05). The cumulative cardiovascular and cerebrovascular event rate in low miR-301a group was 3.70% (4/108), which was lower than that in high miR-301a group [32.04% (33/103), P<0.001]. COX regression analysis showed that high serum albumin level was an independent protective factor for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy,while high hs-CRP level and miR-301a >1.46 were independent risk factors for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. Restricted cubic spline fitting COX regression analysis showed a non-linear relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy (P<0.05). 【Conclusion】 hs-CRP is independently associated with miR-301a in diabetic nephropathy peritoneal dialysis patients. High miR-301a level suggests a high risk of cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy.