Objective Scutellarin (SCU), a Chinese traditional medicine, has a protective effect against ischemia-reperfusion (IR) induced myocardial injury, but it is not yet clear whether SCU acts against vascular endothelial IR injury via extracellular signal-regulated kinase 1/2 (ERK1/2).The aim of this study was to explore the effect of SCU on hypoxia-reoxygenation (HR)-induced injury to human cardiac microvascular endothelial cells (HCMECs) and its influence on the ERK1/2 signaling pathway.Methods HCMECs were subjected to normal culture and divided into a normal control, a DMSO, an SCU 1 μmol/L, and an SCU 10 μmol/L group.The model of HR injury was established by exposing the HCMECs to 12-h hypoxia and 12-h reoxygenation after treated with DMSO or SCU at 1 and 10 μmol/L for 2 hours.Then, the survival rate of the HCMECs was detected by MTT and trypan blue staining, the concentration of malondialdehyde (MDA) in the cells measured, and the expressions of the p-ERK1/2, ERK2 and GAPDH proteins determined by Western blot.Results SCU at 1 and 10 μmol/L significantly increased the survival rate of the normally cultured HCMECs ([110.40±2.34] and [122.00±1.25] ％) as compared with that of the normal control (100％) (P<0.05), while HR injury markedly decreased the vitality of the HCMECs ([68.00±4.06] ％) in comparison with that of the blank control (100％) (P<0.05).The survival rate of the HCMECs was remarkably higher in the HR+SCU 1 μmol/L and HR+SCU 10 μmol/L groups than in the HR model group ([90.53±3.67] and [92.04±2.32] ％) (P<0.05), and so was their vitality in the SCU 10 μmol/L group than in the normal control ([96.78±2.01] vs [90.06±1.85] ％, P<0.01), while their survival rate was significantly lower in the HR model than in the blank control ([73.72±4.91] vs [91.83±2.34] ％, P<0.01) and remarkably higher in the SCU 10 μmol/L ([87.59±2.64] ％) than in the HR model group (P<0.05).The MDA concentration in the HCMECs was markedly increased in the HR model and HR+DMSO groups as compared with the blank control (P<0.01), but decreased in the HR+SCU 1 μmol/L and HR+SCU 10 μmol/L groups in comparison with the HR model group (P<0.05).The expression of the p-ERK1/2 protein was significantly down-regulated in the HR model group as compared with the blank control (P<0.01), but up-regulated in the HR+SCU 10 μmol/L group in comparison with the HR model (P<0.01).Conclusion HR injury reduces the vitality of HCMECs, increases the MDA concentration, and down-regulates the expression of the p-ERK1/2 protein in HCMECs, while SCU acts against ischemia-reperfusion injury to HCMECs by increasing ERK phosphorylation.

Objective To investigate the changes and significances of the level of serum Fibulin-3 and correlation between Fibulin-3 and branchial-ankle Pulse Wave Velocity (baPWV) in patients with hypertension. Methods A total of 50 patients with hypertension and 38 healthy people were enrolled finally, whose fasting blood were collected. The levels of Fibulin-3、MMP-9 were detected by enzyme-linked immunosorbent assay. The baPWV was detected by pulse wave velocity automatic analyzer. Results The level of serum Fibulin-3 in hypertension was lower than the control group significantly (P < 0.001). There was no significance of serum MMP-9 in the two groups. The baPWV in hypertension was higher than the control group.Correlations between the level of Fibulin-3 and baPWV were significant (r = -0.324,P = 0.006). While correlations between the level of MMP-9 and baPWV had no significance (r=0.003,P=0.968). Conclusion The level of serum Fibulin-3 and baPWV can be used as index of functional changes in the early stage of hypertensive vascular remodeling, and may be applied for forecasting the organ damage in hypertension.

Background Bevacizumab has been widely used in the treatment of new blood vessel disease in ophthalmology.The investigation of the pharmacokinetics and safety after intracameral injection of bevacizumab can offer the basis for the management of iris neovascularization and neovascular glaucoma.Objective The present study was to observe the distribution of bevacizumab(avastin)in eye tissue and toxic effects following the injection of anterior chamber.Methods Twenty-four New Zealand albino rabbits were divided into two groups randomly.0.05 ml (1.25mg)of Bevacizumab was intracamerally injected into the left eyes in the experimental group,and a balanced salt solution of 0.05 ml was injected in the same way into the left eyes of the control group.The anterior segment of eyes and ocular fundus were examined by slit-lamp microscope and direct ophthalmoscope after injection.Intraocular pressure was measured and corneal endothelial microscopy was performed before and after the injections.Five rabbits of the two groups were sacrificed on the first day,the fourth day,the seventh day,the fourteenth day,and the thirtieth day after injection,and the eyeballs were enucleated for histopathological examination.The ultrastructure of eye tissue was observed under the transmission electron microscope on the fourth day and the thirtieth day,and then immunofluorescence staining were performed to assess the distribution of bevacizumab in the eye tissues.This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission(Version 1988).Results No abnormality in the cornea,lens,vitreous and retina was observed after the injection of bevacizumab under the slit lamp microscope and direct ophthalmoscope.No significant differences were found in intraocular pressure and corneal endothelial cell density in the bevacizumab group compared with the control group before injection and 2 hours,1 day,7 days,14 days,30 days after injection(P =0.760,P =0.956).No histopathological and ultrastructural changes of the cornea,lens,chamber angle,iris,ciliary body and retina were seen after the injection in the experimental group and control group under the light microscope and transmission electron microscope.Bevacizumab was distributed in the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes and fellow eyes after intracameral injection with red fluorescence and presented the dynamic changes with the lapse of time.The immunofluorescence response of eye tissue to bevacizumab was weaker in the fellow eyes compared with injected eyes.Bevacizumab was mainly distributed in the vessel wall and lumen.Conclusions Bevacizumab can quickly distribute in the vascular tissue of the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes after intracameral injection without obvious toxic effects to eye tissue.Bevacizumab administered intracamerally may be a new strategy or a joint strategy for iris neovascularisation.

Objective To study prevention and treatment for patients during the intermittent period of maintaining hemodialysis complicated by acute left heart failure. Methods The treating process of 126 examples suffered from acute left heart failure during the intermittent period of 14 225 person-time maintaining hemodialysis in 143 patients were retrospectively analyzed. Results In this case group, 0.89% suffered from acute left heart failure during the intermittent period of hemodialysis and the successful rescuing rate was 88.9%. Conclusions Applying the treatment of emergency hemodialysis, quickly filtering body fluid, the curative effect is reliable and rapid. It is one of the best ways to cure patients suffered from acute left heart failure during the intermittent period of maintaining hemodialysis. Combined with other treatments, it will be able to improve the successful rate, and ensure the sufficiency of hemodialysis. United with other steps of reducing blood pressure, such as using angiotensin converting enzyme inhibitors, angiotensin II receptor blocker, and ? receptor blocker, it is an effective countermeasure to prevent patients suffering from acute left heart failure during the intermittent period of maintaining hemodialysis, and effectively reduces its appearance as well.

This article dealed with the effects of processing method and duration on the major bioactive components (sinigrin and sinapine thiocyanate) in Brassica juncea. The contents of sinigrin and sinapine thiocyanate in decoctions of raw and processed B. juncea were determined and compared by high performance liquid chromatography on a Alltima C18 column (4.6 mm x 250 mm, 5 microm) at 35 degrees C with the acetonitrile-0.1% phosphoric acid as the mobile phrase in gradient elution. The detection wavelength of sinigrin and sinapine thiocyanate was set at 227 nm and 326 nm, and the flow rate was 1.0 mL x min(-1). It was found that with the extended processing duration, the contents of sinigrin and sinapine thiocyanate first increased and then decreased: i.e., 0-2 minutes they increased gradually (for sinigrin, by 9.65% in processed products and 356. 10% in powder; for sinapine thiocyanate, by 12.82% in processed products and 3.41% in powder), and achieved their highest content at 2 min; then, decreased during the next 5 minutes (for sinigrin, by 80.35% in processed products and 82.09% in powder; for sinapine thiocyanate, by 14.29% in processed products and 17.54% in powder), suggesting that processing duration could significantly affect the contents of bioactive components in B. juncea, enzymatic hydrolysis of sinigrin when the seed is crushed in the present of moisture may be responsible for the content change. It is recommended that the slow fire should be the best processing method and the raw seed could be used directly in the water extracts related industrial production.
Brassica ; chemistry ; Choline ; analogs & derivatives ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Glucosinolates ; chemistry ; Powders ; chemistry ; Thiocyanates ; chemistry

UNLABELLEDOBJECTIVE To explore the effect of combined use of rehmannia (RM) and rhodiola (RD) on peripheral leukopenia and bone marrow hematopoietic function suppression induced by cyclophosphamide (CTX) in mice.

METHODSICR mice were established into bone marrow inhibition models by intraperitoneal injection of CTX, and were administered with RM, RD or its extract (RDE), singly or in mixture, via gastrogavage for 10 days. The changes of peripheral hemogram, bone marrow nucleated cell proliferation, CFU-GM colony formation, GM-CSF and erythropoietin (EPO) secretion were observed.

RESULTSCompared with the un-treated model mice, the peripheral white blood cell count was significantly higher in model mice treated with RDE and RM mixture; the bone marrow nucleated cells count, CFU-GM formation, and GM-CSF production were significant higher in model mice treated with RD and RM mixture, showing statistical significance (P < 0.01); while EPO production in the RD and RM mixture treated group was slightly elevated, but the difference showed no statistical significance.

CONCLUSIONRD and RM mixture could regulate hematopoietic system by promoting the production of bone marrow cells and colonies, as well as enhancing the synthesis of related cytokines, such as GM-CSF, so as to increase the amount of peripheral white blood cells and restore the hematopoietic function of organism.

Animals ; Bone Marrow ; drug effects ; Cyclophosphamide ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Erythropoietin ; secretion ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; secretion ; Hematopoiesis ; drug effects ; Leukopenia ; chemically induced ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Rehmannia ; chemistry ; Rhodiola ; chemistry

ObjectiveTo investigate the inhibitory effect of bone morphogenetic protein 7 (BMP7) on proliferation of rat hepatic stellate cells (HSC).MethodsHSC-T6 cells were cultured in vitro,and divided into four groups:control group,transforming growth factor (TGF) β1 group,TGFβ1 + BMP7 group,and TGFβ1 + BMP7 + Noggin group. The mRNA expressions of smad1,α-smooth muscle actin (α-SMA),gremlin and fibronectin (FN) were determined by semi-quantitative reverse transcriptasepolymerasechainreaction(RT-PCR).Theproteinexpressionsof phosphorylated-smad1/5/8 (P-smad1/5/8),α-SMA and gremlin were detected by Western blot.The statistical analysis were done using one-factor analysis of variance,LSD-t test,Dunnett's T3 test and Pearson linear correlation analysis.ResultsPhosphorylated-smad1/5/8,α-SMA,gremlin and FN were expressed on HSC-T6 cells in control group,which were significantly up-regulated after TGFβ1 stimulation (P＜0.05).The expressions of α-SMA,gremlin and FN were significantly down-regulated in TGFβ1 + BMP7 group compared with TGFβ1group, while P-smad1/5/8 expression was upregulated (1.613±0.031 vs.2.503±0.014; t=34.89,P＜0.05).The expressions of α-SMA,gremlin and FN in TGFβ1 + BMP7 + Noggin group were up-regulated than those in TGFβ1 + BMP7 group,while P-smad1/5/8 was down-regulated (t=34.05,P＜0.05).There was no difference of total smad1 mRNA expression among each group.ConclusionThese results collectively suggest that BMP7 strongly inhibits the expressions of α-SMA,gremlin and FN.

Objective To explore the dynamic expressions of interleukin-1 (IL-1)receptor associated kinase (IRAK)-M and IRAK-4 in rats with or without endotoxin tolerance (ETT)in acute liver failure (ALF).Methods Sixty-six male SD rats were divided into three groups:ALF group,ETT group and control group.The rats in ETT group received daily lipopolysaccharide (LPS)intraperitoneal injection for 5 days,while the rats in ALF group received daily injection with same volume of 0.75% NaCl solution.Both ETT group and ALF group received intraperitoneal injection with D-galactosamine (D-GalN)and LPS at 24 hours after the 5th injection of LPS or NaCl solution.At 2,6,12,24 and 48 h after D-GalN and LPS injection,the serum levels of tumor necrosis factor-α (TNF-α)and interleukin-6 (IL-6)were detected by enzyme-linked immunosorbent assay (ELISA).The liver pathologic changes were observed with HE staining by microscope.The mRNA expressions of IRAK-4,IRAK-M and NF-κB (p65)were detected by reverse transcriptase polymerase chain reaction (RT-PCR).The pairwise comparison between groups was done by lease significant difference (LSD)and Dunnet's t test.Results The liver pathologic changes in ETT group were much milder than those of ALF group.In ALF group,IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24and 48 h after D-GalN and LPS challenge were 0.711 ±0.074,0.904±0.118,1.012 ±0.098,1.534±0.279 and 1.451±0.290,respectively,while the IRAK-M mRNA/β-actin absorbance ratios were 0.496±0.018,0.516±0.089,0.503±0.023,0.503±0.057 and 0.469±0.142,respectively.In ETT group,the IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24 and 48 h after D-GalN and LPS challenge were 0.619±0.083,0.587±0.033,0.623±0.034,0.720±0.044 and 0.654±0.041,respectively,while the IRA'K-M mRNA/β-actin absorbance ratios were 0.929 ± 0.064,1.111±0.138,1.113±0.027,1.891±0.315 and 1.710±0.303,respectively.The IRAK-M mRNA expression level was significantly increased and IRAK-4 mRNA expression level was relatively decreased in ETT group compared to ALF group.The differences were all statistically significant at 2,6,12,24 and 48 h after D-GalN and LPS challenge (F= 17.305,54.921,121.031,67.607,55.279,respectively; F=19.506,43.777,110.823,302.681,202.822,respectively; F=172.003,59.519,987.055,68.463,96.601,respectively; all P＜0.05).Conclusion LPS pretreatment may induce ETT in rat model by downregulating the expression of IRAK-4 and upregulating the expression of IRAK-M.

Objective To observe the expression of axon guidance cues Slit2 and Robo4 in lung tissue of rat with acute lung injury (ALI) and explore the function of Slit2 and Robo4 in ALI.Methods Forty-eight Sprague-Dawley rats were randomly (random number) divided into control group (n =24) and ALl group (n =24).ALI model was reproduced by cecum ligation and puncture (CLP).The control group only experienced a simulated operation without CLP.Both groups were further divided into 3 subgroups with 8 rats in each subgroup:12 h,24 h,and 48 h subgroups.artery blood gas analysis,lung tissue wet/dry weight (W/D) ratio,lung histopathologic changes,pulmonary microvascular permeability were observed.The serum tumor nocrosis factor-α (TNF-α) was measured with enzyme linked immunosorbent assay (ELISA).The expression of Slit2 and Robo4 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expression of Slit2 and Robo4 protein in lung tissues was assessed by immunohistochemistry.Date were analyzed by one-way ANOVA with SPSS version 13.0 software.Statistical significance was established at a P value of less than 0.05.Results Compared with the control group,in ALI rats at different time points,partial pressure of oxygen in arterial blood (PaO2) decreased significantly,lung W/D weight ratio and pulmonary microvascular permeability,the serum TNF-α increased significantly (all P ＜ 0.05),histopathology of lung revealed signs of injury.The expression of Slit2 mRNA in lung tissues was decreased markedly after CLP compared with control group [(0.56±0.13) vs.(0.87±0.05),F=41.39,P＜0.05,(0.42±0.10) vs.(0.85±0.07),F=93.54,P＜0.05,(0.26±0.08) vs.(0.89 ±0.09),F=227.05,P＜0.05].but there were no significant difference in expression of Robo4 mRNA in lung tissue between ALI group and control group [(0.86±0.07) vs.(0.83±0.05),F=0.695,P＞0.05,(0.82±0.05) vs.(0.89±0.08),F=2.061,P ＞ 0.05,(0.86 ± 0.08) vs.(0.86 ± 0.05),F =0.035,P ＞ 0.05].Immunohistochemistry study showed Slit2 protein was mainly expressed on the extracellular surface of vascular endothelial cells,while lung epithelial cell nuclei and endochylema.Robo4 protein was only expressed on the extracellular surface of vascular endothelial cells.Compared with the control group,expression of Slit2 protein in lung tissue in ALI group decreased markedly [(0.37 ± 0.05) vs.(0.45 ± 0.07),F =6.82,P ＜ 0.05,(0.32±0.06) vs.(0.47±0.09),F=23.54,P＜0.05,(0.28±0.07) vs.(0.46±0.06),F=28.01,P ＜ 0.05].As good as RT-PCR,there were no significant difference in expression of Robo4 protein in lung tissue between two groups [(0.53±0.04) vs.(0.52±0.05),F=0.155,P＞0.05,(0.53± 0.09) vs.(0.50±0.05),F=0.498,P＞0.05,(0.55±0.06) vs.(0.56±0.07),F=0.073,P ＞ 0.05].Conclusions Lung tissues of control group rats express Slit2 and Robo4.The decreased Slit2 mRNA and protein expressions in the lung tissue of rat with ALI caused by CLP may be associated with the occurrence of ALI.

Objective To assess the curative effect of endoscopic resection for patients with duode-nal bulb carcinoid.Methods Data of 17 patients with duodenal bulb submucosal tumor who underwent en-doscopic dissection in our department and confirmed as duodenal bulb carcinoid by postoperative pathology from Jun 2009 to Jun 2012 were retrospectively analyzed.Seventeen patients included 1 1 men and 6 women with the mean age of 36. 3 ±8. 4.Results All patients underwent preoperative diagnosis of endoscopic ultrasonography(EUS).Four cases were diagnosed as heterotopic pancreas and 13 cases carcinoid.The size of tumor was from 4 to 10 millimeter.Cap-assisted endoscopic mucosal resection (EMR-C)were used in all patients successfully.No complications were found during or after the operation.Postoperative pathology con-firmed 6 cases of duodenal bulb carcinoid.The accuracy of EUS preoperative diagnosis was 76. 5%.The average follow-up time was 20. 5 ±12. 4 months.Metastases and recurrence had not been found.Conclusion EUS can confirm the invasive depth of duodenal submucosal tumors and estimate the indication of endo-scopic excision.EUS can not give a preoperative qualitative diagnosis of duodenal submucosal tumors.Endo-scopic hyaline cap excision is a safe and sufficient method for duodenal bulb carcinoid.