GDF11 ameliorated myocardial ischemia reperfusion injury by antioxidant stress and up-regulating autophagy in STZ-induced type 1 diabetic rats

Abstract Purpose: To investigate whether GDF11 ameliorates myocardial ischemia reperfusion (MIR) injury in diabetic rats and explore the underlying mechanisms. Methods: Diabetic and non-diabetic rats subjected to MIR (30 min of coronary artery occlusion followed by 120 min of reperfusion) with/without GDF11 pretreatment. Cardiac function, myocardial infarct size, creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) 15-F2tisoprostane, autophagosome, LC3II/I ratio and Belcin-1 level were determined to reflect myocardial injury, oxidative stress and autophagy, respectively. In in vitro study, H9c2 cells cultured in high glucose (HG, 30mM) suffered hypoxia reoxygenation (HR) with/without GDF11, hydrogen peroxide (H2O2) and autophagy inhibitor 3-methyladenine (3-MA) treatment, cell injury; oxidative stress and autophagy were assessed. Results: Pretreatment with GDF11 significantly improved cardiac morphology and function in diabetes, concomitant with decreased arrhythmia severity, infarct size, CK-MB, LDH and 15-F2tisoprostane release, increased SOD activity and autophagy level. In addition, GDF11 notably reduced HR injury in H9c2 cells with HG exposure, accompanied by oxidative stress reduction and autophagy up-regulation. However, those effects were completely reversed by H2O2 and 3-MA. Conclusion: GDF11 can provide protection against MIR injury in diabetic rats, and is implicated in antioxidant stress and autophagy up-regulation.

Experimental study of allicin inhibiting the proliferation and invasion of human multiple myeloma cell / 国际肿瘤学杂志

Objective To investigate the effect of allicin on the proliferation,apoptosis and invasion of human multiple myeloma (MM) cell line RPMI8226 in vitro,and to explore its mechanism.Methods The human MM cell line RPMI8226 cells were treated with different concentrations of allicin as 0,5,25,125 μmol/L,wherein the control group was 0 μmol/L allicin treatment group.The proliferation of cells was calculated by cell counting kit-8 (CCK-8).The apoptosis and invasion ability of tumor cells were determined with flow cytometry and Transwell method respectively.The expressions of transforming growth factor-α (TGF-α),matrix metalloproteinase (MMP)-2,MMP-9 and tissue inhibitor of metalloproteinase-2 (TIMP-2)protein were detected by Western blotting.Results The proliferation rates of RPMI8226 cells in groups treated with 0,5,25 and 125 μmol/L allicin were 1.00％ ± 0.02％,0.98％ ± 0.04％,0.73％ ± 0.22％ and 0.44％ ± 0.15％ respectively,with a significant difference (F =329.2,P ＜ 0.001).The proliferation rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly inhibited compared with control group (P ＜0.001;P ＜0.001);but there was no significant difference between 5 μmol/L treatment group and control group (P=0.395).The apoptosis rates of RPMI8226 cells treated with 0,5,25 and 125 μmol/L allicin were 3.05％ ±0.53％,4.06％ ±0.29％,12.17％ ± 1.08％ and 12.81％ ± 1.78％ respectively,with a significant difference (F =531.0,P ＜0.001).The apoptotic rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly increased compared with control group (P =0.013;P =0.012);and there was no significant difference between 5 μmol/L treatment group and control group (P =0.211).The numbers of invasive cells in the 0,5,25 and 125 μmol/L allicin treatment groups were 112.5 ± 1.9,104.8 ±4.0,76.9 ± 2.6 and 52.5 ± 3.7 respectively,with a significant difference (F =734.9,P ＜ 0.001).The abilities of cells invasiveness in 25 and 125 μmol/L allicin treatment groups were significantly decreased compared with control group (P ＜ 0.001;P ＜ 0.001),but 5 μmol/L allicin treatment group had no significant difference compared with the control group (P =0.160).Western blotting results showed that the expression levels of TGF-α,MMP-2,MMP-9 and TIMP-2 proteins were significantly different between groups treated with different concentrations of allicin (F =227.1,P＜0.001;F=348.5,P＜0.001;F=359.7,P＜0.001;F=158.0,P ＜0.001).The protein expression levels of TGF-eα,MMP-2 and MMP-9 were significantly decreased in the 25 and 125 μmol/L treatment groups compared with the control group (all P ＜ 0.001),and the expression of TIMP-2 protein was significantly increased compared with the control group (all P ＜ 0.001),but there were no significant diffe-rences between the 5 μmol/L treatment group and the control group (P =0.349;P =0.744;P =0.613;P =0.567).Conclusion Allicin can significantly inhibit cell proliferation,invasive ability and induce apoptosis of human MM cell line RPMI8226 in vitro,and the mechanism may be related to inhibiting the expressions of TGF-α,MMP-2,MMP-9 and promoting the expression of TIMP-2 at the same time.

Mechanism underlying docosahexaenoic acid-induced regulation of angiopoietin expression in human brain vascular pericytes subjected to oxygen-glucose deprivation and restoration: the relationship with SSeCKS / 中华麻醉学杂志

Objective To evaluate the relationship between the mechanism underlying docosahexaenoic acid (DHA)-induced regulation of angiopoietin expression and Src-suppressed C kinase substrate (SSeCKS) in human brain vascular pericytes (HBVPs) subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods HBVPs were seeded in 96-well or in 6-well plates at a density of 2× 105 cells/ml and divided into 5 groups (n =18 each) using a random number table:control group (group C),OGD/R group,DHA group (group D),SSeCKS gene silencing group (group S) and SSeCKS gene silencing plus DHA group (group SD).The model of OGD/R injury was established as follows:the cells were subjected to O2-glucose deprivation for 24 h in glucose-and serum-free culture medium aerated with 94％ N2-5％ CO2-1％ O2 followed by restoration of O2-glucose supply for 6 h in high-glucose DMEM culture medium in normal atmosphere.DHA was added at 1 h before hypoxia with the final concentration of 40 μmol/L in group D.Small interfering RNA induced SSeCKS gene silencing in S and SD groups.Subsequently,DHA with the final concentration of 40 μmol/L was added at 1 h before hypoxia in group SD.At 6 h of reoxygenation,the cell survival rate was determined by CCK-8 assay,the amount of LDH released was detected using ELISA,and the expression of SSeCKS,angiopoietin-1 (Ang-1) and Ang-2 was detected by Western blot.Results Compared with group C,the cell survival rate was significantly decreased,the amount of LDH released was increased,the expression of SSeCKS and Ang-1 was down-regulated,the expression of Ang-2 was up-regulated,and Ang-1/Ang-2 ratio was decreased in group OGD/R,and the expression of SSeCKS was down-regulated in group S (P＜0.05).Compared with group OGD/R,the cell survival rate was significantly increased,the amount of LDH released was decreased,the expression of SSeCKS and Ang-1 was up-regulated,the expression of Ang-2 was down-regulated,and Ang-1/Ang-2 ratio was increased in group D (P＜O.05),and no significant change was found in the parameters mentioned above in group SD (P＞0.05).Compared with group D,the cell survival rate was significantly decreased,the amount of LDH released was increased,the expression of SSeCKS and Ang-1 was down-regulated,the expression of Ang-2 was up-regulated,and Ang-1/Ang-2 ratio was decreased in group SD (P＜0.05).Conclusion The mechanism by which DHA increases the ratio of Ang-1/Ang-2 may be totally related to up-regulation of SSeCKS expression in HBVPs subjected to OGD/R.

Effect of Weide'an Tablet on Gastric Function and the Expressions of EGF and EGFR in Gastric Tissue of Model Rats with Stress-induced Gastric Injury / 中国药房

OBJECTIVE:To study the effects Weide'an tablet on gastric function and the expressions of epidermal growth fac-tor(EGF)and its receptor(EGFR)in gastric tissue of model rats with stress-induced gastric injury,and investigate its mechanism in the treatment of gastric ulcer. METHODS:Rats were randomly divided into normal group (normal saline),model group (nor-mal saline),positive group (ranitidine hydrochloride,0.015 mg/kg) and Weide'an tablet high-dose,medium-dose,low-dose groups (1.7,0.87,0.43 g/kg),10 in each group. Except for normal group,rats in other groups were given fasting swimming in cold water to induce gastric ulcer model. After modeling,all rats were intragastrically administrated once a day,for 2 weeks. The body mass of rats in 0,7,14 d of administration was recorded. After 12 h of last administration,gastric juice secretion,gastric juice pH,gastric ulcer area and gastric mucosal damage index of rats were detected,and the expressions of EGF and EGFR in gas-tric tissue were detected. RESULTS:Compared with normal group,body mass and gastric juice pH in 7,14 d in model group were declined;gastric juice secretion and gastric ulcer area were increased,gastric mucosal damage index was increased;and the expressions of EGF and EGFR in gastric tissue were enhanced,with statistical significances(P<0.05 or P<0.01). Compared with model group,except that the body mass,gastric juice secretion,gastric juice pH and gastric ulcer area in Weide'an medium-dose group in 7 d,and body mass,gastric juice secretion,gastric juice pH,gastric ulcer area and the expression of EGF in gastric tis-sue in Weide'an low-dose group in 7,14 d were not significantly improved,above indexes were significantly improved in other ad-ministration groups(P<0.05 or P<0.01);and the effect of Weide'an tablet showed a certain dose-effect relationship. CONCLU-SIONS:Weide'an tablet can significantly improve the gastric function of model rats with stress-induced gastric injury;and the mechanism may be related with enhancing the expressions of EGF and EGFR and promoting ulcer healing.

Identification and determination of scutellarin in compound granule tetrandra / 药学实践杂志

Objective To develop the methods for identification and determination of scutellarin in compound Granule Tetrandra .Methods A TLC method was used to identify quality for scutellarin ,and a HPLC method was used to determine the content of scutellarin .Results The spot was clear in TLC identification without interference of negative control .The sam-ple size of scutellarin had a good linear relationship with peak area r=0 .9996(n=5)when ranged from 22 .4~156 .8 μg/ml , with the average recovery rate 97 .79% ( RSD=0 .32% ,n=6) .Conclusion The method is simple ,with good specificity and reproducibility ,which can be used as the quality control for this preparation .

Effects of docosahexaenoic acid on expression of Ang-2 and VEGF in rat brain microvascular endo-thelial cells subjected to oxygen-glucose deprivation / 中华麻醉学杂志

Objective To evaluate the effects of docosahexaenoic acid ( DHA) on the expression of angiotensin?2 ( Ang?2) and vascular endothelial growth factor ( VEGF) in rat brain microvascular endo?thelial cells (BMVECs) subjected to oxygen?glucose deprivation (OGD). Methods Primarily cultured rat BMVECs were divided into 3 groups ( n=18 each) using a random number table: control group ( group C) , OGD group and DHA group. The cells were cultured with glucose?free and serum?free DMEM culture medium in OGD and DHA groups. In group DHA, DHA 40μmol was added, and then the cells were ex?posed to 1%O2?5%CO2?94%N2 in an air?tight incubator. The cells were cultured in the normal glucose and normal oxygen conditions in group C. All the cells were cultured for 24 h. Cell apoptosis was detected using Annexin V∕propidium iodide flow cytometry assay, and the apoptosis rate was calculated. The concentra?tions of Ang?2, VEGF, prostaglandin E2 ( PGE2 ) and prostacyclin ( PGI2 ) in the supernatant of the cul?ture medium were determined by enzyme?linked immunosorbent assay. The expression of Ang?2 mRNA and VEGF mRNA in cells was detected by real?time polymerase chain reaction. The expression of cyclooxygen?ase?2 ( COX?2) protein in cells was detected by Western blot. Results Compared with group C, the ap? optosis rate and concentrations of Ang?2, VEGF, PGE2 and PGI2 in the supernatant were significantly in?creased, and the expression of Ang?2 mRNA, VEGF mRNA and COX?2 protein was significantly up?regu?lated in OGD and DHA groups (P<0.05). Compared with group OGD, the apoptosis rate and concentra?tions of Ang?2, VEGF, PGE2 and PGI2 in the supernatant were significantly decreased, and the expression of Ang?2 mRNA, VEGF mRNA and COX?2 protein was significantly down?regulated in group DHA ( P<0.05) . Conclusion DHA can inhibit the expression of Ang?2 and VEGF in rat brain BMVECs subjected to OGD and reduce cell apoptosis, and down?regulated expression of COX?2 protein is involved in the mecha?nism.

Effects of Oscillator on the Cleanliness of Class 100 Clean Bench in Pharmacy Intravenous Admixture Ser-vices / 中国药房

OBJECTIVE:To study the influence of oscillator on the cleanliness of class 100 clean bench in PIVAS. METH-ODS:Using sedimentated bacteria and the number of dust particle as index,in common drug configuration room,antibiotics con-figuration room and risk drugs configuration room including biological safety cabinet and horizontal laminar flow,the cleanliness of class 100 clean bench were monitored when oscillator was set at clean bench and different positions in work and non-working state. RESULTS & CONCLUSIONS:In working and non-working state of oscillator,there was no difference in sedimentated bacteria and the number of dust particle which was in line with the requirements of 2010 edition of GMP,i.e. the application and location of oscillator didn't influence the cleanliness of class 100 clean bench. From a view of safety,it is suggested to place the oscillator in the left(or right)posterior wall of clean table when biological safety cabinets is used to dispense antibiotic and risk drugs.

Protective Effect of Weidean Tablets on Rats with Stress-induced Gastric Ulceration / 中国药房

OBJECTIVE:To study the protective effect of Weidean tablets on rats with stress-induced gastric ulceration. METH-ODS:Water immersion restraint stress method and empty stomach were employed to establish rat models with gastric ulceration. 60 SD rats were equally randomized into normal control group(isometric sodium chloride injection),model group(isometric sodium chloride injection),ranitidine group(0.015 g/kg),and groups of high,medium and low doses(1.70,0.87 and 0.43 g/kg)of Wei-dean tablets. General situation of stomach was observed .The gastric mucosal injury index,the NO content in serum,and the activi-ties of SOD and iNOS and the contents of MDA and PGE2 in the gastric tissue were detected for rats. Pathomorphological observa-tion of rats’gastric tissues was conducted under the microscope. RESULTS:Compared with normal control group,the rats in the model group had higher gastric mucosal injury index,lower content of NO in serum,and weaker activity of SOD,stronger activity of iNOS,higher content of MDA and PGE2,in the gastric tissue. There was statistical significance (P<0.01 or P<0.05). Dark matter,severe deformation and necrosis of gastric mucosal cells in rats were noted,as well as multiple large-area superficial ulcers. Compared with the model group,the rats in groups of high,medium and low doses of Weidean tablets had lower gastric mucosal injury index,higher content of NO in serum,and stronger activity of SOD,weaker activity of iNOS,lower content of MDA and higher content of PGE2,in the gastric tissue. There was statistical significance(P<0.01 or P<0.05). General situation of stomach and the pathomorphological condition of rats’gastric tissues were improved. CONCLUSIONS:Weidean tablets has protective ef-fect to some extent on rats with stress-induced gastric ulceration,the mechanism is related to the improvement of serum levels of anti-oxidation index in rats.

Effect of Compound Pueraria Tablets on Osteoprosis Model Rats Induced by Retinoic Acid / 中国药房

OBJECTIVE:To study the effect of Compound pueraria tablets on osteoporosis model rats induced by retinoic acid. METHODS:The osteoporosis model was induced by intragastric administration of retinoic acid solution for 15 days;normal group was established. After modeling,the rats were randomly divided into model control group,Xianling gubao capsule [0.32 g/(kg·d)] positive control group,Compound pueraria tablets low-dose and high-dose [0.24,0.4 g/(kg·d)] groups (n=8). After 6 weeks of ig,the serum sample was collected to determine the levels of serum calcium(s-Ca),serum phosphorus(s-P),ALP and bone gla protein (BGP);bone density instrument was used to detect the contents of bone mineral density (BMD),bone mineral content (BMC),bone image area (BIA) and muscle content (MC);the results of compact bone substance scanning were observed. RE-SULTS:Compared with normal group,the levels of s-Ca,BMD,BMC and MC in rats were decreased in model control group, while the level of BGP was increased(P<0.05 or P<0.01). Compared with model control group,related index and compact bone substance scanning of Compound pueraria tablets groups were all improved;the levels of s-Ca,s-P,ALP,BMD and MC were in-creased in Compound pueraria tablets high-dose group,while the level of BGP was decreased;the levels of BMD and MC were in-creased significantly in Compound pueraria tablets low-dose group(P<0.05 or P<0.01). CONCLUSIONS:Compound pueraria tab-lets can improve the osteoporosis induced by retinoic acid in rats.

Study on Protective Effect of Compound Pueraria Tablets on Myocardium of Aging Model Rats Induced by D-galactose / 中国药房

OBJECTIVE:To study the protective effect of Compound pueraria tablets on the aging model rats. METHODS:The rats were divided into normal group,model control group,positive control group (vitamin E) and Compond pueraria tablets high-dose,medium-dose and low-dose (800,400,200 mg/kg) groups. The aging model rats were made by injection of D-galac-tose solution for 30 d in latter 5 groups,and they were given relevant medicine at same time every 7 days for consecutive 56 days since 31st day. SOD activity and MDA content in the serum and myocardium and LPF contents in the myocardium were determined after the last dosing,and apoptotic cells were counted and the pathology of cerebral tissues were observed. RESULTS:Compared with normal group,the activity of SOD in the serum and myocardium were decreased significantly in model control group,MDA and LPF contents in the myocardium and the number of apoptotic cells were increased significantly(P<0.01);significant myocar-dial cell injury was found. Compared with model control group,the activity of SOD in the serum and myocardium were increased significantly,while MDA and LPF content in the myocardium and the number of apoptotic cells in Compound pueraria tablets high-dose,medium-dose and low-dose groups were decreased significantly(P<0.05 or P<0.01);myocardial cell morphology was improved significantly. CONCLUSIONS:Compound pueraria tablets has obvious protective effect on myocardial cells in aging rats induced by D-galactose,the mechanism maybe related to the increase of SOD activity and the decrease of the content of MDA and LPF.

Protective Effect of Compound Pueraria Tablets on the Kidney Tissues of Diabetic Nephropathy Model Rats / 中国药房

OBJECTIVE:To explore the effect of Compound pueraria tablets on the kidney tissues of diabetic nephropathy rats. METHODS:High sugar and lipid diet combined with intraperitoneal injection of streptozotocin were used to establish the model of diabetic nephropathy (DN). Normal control group was established. Model rats were randomly divided into model control group, positive control group(Irbesartan tablet),Compound pueraria tablets low-dose,medium-dose and high-dose [0.102,0.203,0.406 g/(kg·d)] groups(n were 8-10). The corresponding drugs were given,and fasting blood glucose(FBG)and 24 h urinary protein (Upro) were collected at 1st,14th,28th,42nd,56th day after treatment. SCr,BUN,TC,TG and KI were detected,and renal pathology was observed after the last dose. RESULTS:Compared with normal group,FBG of those groups were all increased after modeling,and 24 h Upro of them were all increased after 28th day (P<0.05 or P<0.01). Compared with model control group, FBG of Compound pueraria tablets medium-dose and high-dose groups were all decreased since 42nd day (P<0.05 or P<0.01), and 24 h Upro of Compound pueraria tablets groups were all decreased since 28th day(P<0.05 or P<0.01);BUN,TC,TG and KI of Compound pueraria tablets in medium-dose and high-dose groups were all decreased significantly,and SCr of 3 dose groups were all increased (P<0.05 or P<0.01). The morphological structure of renal cells was improved significantly in drug treatment groups. CONCLUSIONS:Compound pueraria tablets can correct lipid metabolism disorder,reduce Upro and improve renal func-tion,indicating certain protective effect on the kidney tissues of diabetic nephropathy rats.

Behavioral intervention on quality of life in chronic heart congestive failure patients with urinary incontinence: a pilot randomized controlled trial / 中国实用护理杂志

Objective To explore the effect of behavioral intervention on the quality of life of chronic heart congestive failure patients with urinary incontinence, to further provide basis for more effective clinical nursing measures. Methods 62 patients with chronic heart congestive failure combining urinary incontinence were divided into two groups by random digital table method, the control group and the intervention group, with 31 cases in each group. Patients in the control group received routine nursing care, while patients in the intervention group received behavioral intervention on the basis of routine care. The behavioral intervention included pelvic floor muscle training, pectineus exercise and reconstruction of micturition habits. The effect of the intervention on incontinence, quality of life and depression were observed in two groups. Results 61 patients completed the study, including 30 cases in the intervention group and 31 cases in the control group. There were no significant differences in the scores of the International Consultation on Incontinence Questionnaire-Urinary Incontinence Short Form (ICI-Q-SF), the MOS item Short Form Health Survey (SF-36) and Zung Self-rating Depression Scale ( SDS) between two groups before the intervention (P>0.05). After 6 weeks of continuous intervention, the scores of ICI-Q-SF and SDS in the intervention group were(8.69±1.94)points and (55.91±4.57) points, which were significantly lower than (11.07±2.14) points and (61.44±5.98) points of the control group (t=5.04, 3.18, P<0.01). About the SF-36, scores of physical functioning and social functioning in the intervention group were (76.77 ±10.34) points and (77.69±6.17) points, which were significantly higher than (60.39±10.07) points and (59.38±8.25) points in the control group (t=43.31, 120.36, P<0.01). Conclusions For chronic heart congestive failure patients with urinary incontinence, behavioral intervention can effectively ameliorate symptoms of urinary incontinence, improve the quality of life and relieve patients' depression, which thereby potentially promote patients′physical and mental health.

Research in effect of oral cavity nursing on senile patients with diabetes mellitus / 中国实用护理杂志

Objective To discuss oral cavity nursing in senile diabetic patients with oral disease.Methods 80 senile diabetic patients were offered diet control and treatment for lowering blood glucose with simultaneous oral nursing for 4 weeks.The blood glucose,blood lipid before and after nursing were observed.Results The total effective rate reached 91.3% after oral nursing,blood glucose and blood lipid greatly lowered after nursing.Conclusions Prevention of oral disease is important for preventing diabetic complications,appropriate oral nursing contributes to prevention and treatment of diabetes mellitus complicated with oral disease.

Treatment of dissecting aneurysm using aorta stent implantation: Analysis of anesthesia related factors / 中国组织工程研究

The disease incidence of aortic dissecting aneurysm showed an increasing tendency.Recently,the clinical application of endovascular stent-graft implantation received good results in treating aortic dissecting aneurysm,which is characterized by minimally invasive,safe,with high success rates.However,the chosen of anesthesia is still in dispute.Local anesthesia,intraspinal anesthesia and general anesthesia can be used in this operation.The investigation demonstrated that intubation general anesthesia is more security in this operation with less complication,which is conductive to controlling blood pressure and braking conditions.

Preparation and Quality Control of Breviscapine Orally Disintegrating Tablet / 中国药房

OBJICTIVE: To prepare Breviscapine orally disintegrating tablet and to establish its quality control method. METHODS: The formula of the core of the tablet was optimized with disintegration time as a parameter by an orthogonal design, and the content and the related substances of Breviscapine orally disintegrating tablet were determined by HPLC. RESULTS: The prepared Breviscapine orally disintegrating tablet showed a good stability, and its characters, dissolution, content and the related substances were all in conformity with the related specifications stated in Chinese Pharmacopoeia (2005 edition). CONCLUSION: The methods of preparation and Quality control are simple and feasible.