OBJECTIVETo investigate the effect of Xianxiong decoction on the mice with acute lung injury induced by lipopolysaccharide.

METHODEighty female ICR mice were randomly divided into 8 groups: model group, Xianxiong decoction group, Daxianxiong decoction group, Xianxiong decoction group without Kansui Radix group, Xianxiong decoction group without Glycyrrhizae Radix et Rhizoma group, Glycyrrhizae Radix et Rhizoma and Kansui Radix group, normal group and control group. Animals of each group, except normal group, were undertaken intraperitoneal injection and intranasal inhalation of lipopolysaccharide (LPS) on day 1, 2, 3 to establish acute lung injury (ALI) model. 30 min after modeling, 0.2 mL corresponding drugs were administrated to each mice, dexam ethasone and normal saline were given to the mice of control group and normal group respectively. White blood cell in blood, neutrophil percentage of blood and bronchoalveolar lavage fluid (BALF) supernatant, the ratio of wet and dry lung tissue ( W/D), histopathological changes of lung tissue were estimated. Sixty ICR mice were randomly divided into normal, model, control, high, middle and low dose Xianxiong decoction groups and were modeled in the same way. ELISA was applied to detect the level of NF-kappaB, TNF-alpha and IL-6 in BALF, PCR for NF-kappaB and TNF-alpha mRNA in lung tissue, and Western blot for NF-kappaB and TNF-alpha. Half of 20 ICR mice were administrated with Xianxiong decoction of its maximum tolerant normal saline.

RESULTCompared with model group, the number of WBC in blood of Xianxiong decoction group mice decreased (P < 0.01), percentage of neutrophils in both blood and BALF decreased as well (P < 0.01, P < 0.05); it also significantly reduced the ratio of W/D (P < 0.01); and found the alveolar wall, the number of inflammatory cells infiltrating improved, compared with model group. Xianxiong decoction reduced the level of NF-kappaB, TNF-alpha and IL-6 in BALF (P < 0.01, P < 0.01, P < 0.05); its high and low dose groups only found TNF-alpha level declined. Five mice died 24 h after administration of Xianxiong decoction which indicated its toxicity when other influential factors were considered.

CONCLUSIONXianxiong decoction is effective on the ALI mice induced by LPS, but it is of toxicity at 3 g x mL(-1).

Acute Lung Injury ; drug therapy ; genetics ; metabolism ; pathology ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred ICR ; NF-kappa B ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Objective To investigate the influence of diabetes liaisons on the practicing skills of nurses and life quality of patients in neurology department.Methods Diabetes liaisons were established in 2009 in neurology department.40 nurses' practicing skills of diabetes nursing were compared 3 months before with 3 months after the establishment,and 120 diabetes patients' knowledge 3 months before and 112 diabetes patients' knowledge 3 months after the establishment were compared.Results The correct rates of 40 nurses' practicing skills of diabetes nursing in blood sugar monitoring,insulin pen injection and hypoglycemic emergency measures were 70％,75％ and 60％ 3 months before the establishment of diabetes liaisons and 100％,100％,and 100％ 3 months after the establishment of diabetes liaisons.And the differences were statistically significant (x2 =69.83,72.57,60.28,respectively;P ＜0.01).The rates of diabetes patients' knowledge in the field of reasonable diets,drug knowledge,regular exercises,skin care and self management were respectively 76.79％,64.07％,59.82％,58.04％ and 82.14％ 3 months after and 29.17％,20.83％,33.33％,20.00％ and33.33％ 3 months before the establishment.And differences were statistically significant (x2 =44.68,46.28,13.33,27.73,47.64,respectively; P ＜ 0.01).Conclusions Diabetes liaisons in the department of neurology can improve nurse' s practicing skills and both nurse' s and patient' s knowledge of diabetes,so as to improve nurse' s nursing quality as well as patient' s life quality.

AIM:To investigate the effects of evodiamine on the growth and apoptosis of human hepatocellular carcinoma Huh7 cells,and to illustrate the molecular mechanism that evodiamine enhances antitumor activity of tumors nec -rosis factor-related apoptosis-inducing ligand(TRAIL)in Huh7 cells.METHODS: The cell viability was measured by MTT assay.The cell cycle distribution was analyzed by flow cytometry.The apoptosis rate was determined by TUNEL stai-ning.The protein levels of cell cycle-and apoptosis-related proteins were detected by Western blot analysis.RESULTS:Treatment of Huh7 cells with evodiamine reduced the cell viability(P<0.05).Evodiamine induced cell cycle arrest in G2/M phase by upregulation of p27,cyclin B1, cell division cycle protein 2(Cdc2)and p-Cdc2.Evodiamine triggered apoptosis accompanied by cleavage of caspase-3 and poly(ADP-ribose)polymerase(PARP).Combination of evodiamine with TRAIL significantly reduced the cell viability and increased cleavage of caspase -3 and PARP as compared with the use of each agent alone.Moreover,evodiamine increased the expression of death receptor 5(DR5)in the Huh7 cells.CON-CLUSION:Evodiamine inhibits the cell growth by reducing the cell viability and inducing cell cycle arrest.Evodiamine also triggers cell apoptosis and enhances the sensitivity of Huh 7 cells to TRAIL by upregulating the expression of DR5.

OBJECTIVETo study the effects of lipid rafts on cell signal transmembrane transduction mediated by c-Met.

METHODSAfter HepG2Cells were treated with MbCD to disrupt the lipid rafts and were treated with artificial recombination hepatocyte growth factor to activate c-Met, the activities of PLCr1/PKC, PI3K/Akt and MAPK signaling pathways in HepG2 cells were analyzed using Western blot.

RESULTS(1) After disruption of lipid rafts with MbCD, phosphorylation of PLCr1 decreased by 35% (P = 0.022); the content of PLCr in the cytoplasm increased by 1.75 fold (P = 0.017); PLCr1 conjugated with membrane decreased by 30% (P = 0.037). (2) The content of PKB in the cytosol decreased by 38% (P = 0.028), and the phosphorylation level of PKB conjugated with membrane decreased by 14% (P = 0.041). At the same time, PDK translocation from cytosol to the plasma membrane and its activation were inhibited by treatment with MbCD. (3) Treatment with MbCD had no significant effect on ErK/MAPK, p38/MAPK and JNK/MAPK signaling pathways.

CONCLUSIONDisruption of lipid rafts with MbCD inhibits the activation of PLCr1/PKC and PI3K/PKB signaling pathways by HGF/cMet, but has no effect on MAPK signaling pathway.

Hep G2 Cells ; Humans ; Membrane Microdomains ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phospholipase C gamma ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-met ; metabolism ; Signal Transduction

Toxicity of different processed was evaluated Polygoni Multiflori Radix by determining the hepatotoxic potency for selecting processing technology. Process Polygoni Multiflori Radix using high pressure steamed, Black Bean high pressure steamed, atmospheric steamed for different time. Using normal human hepatocytes (L02) as evaluation model, hepatotoxic potency as index to evaluate hepatotoxic potency of different processed Polygoni Multiflori Radix. Analysis chemical composition of some processed products by UPLC-MS. Hepatotoxic bioassay method cloud evaluate the toxicity of different Polygoni Multiflori Radix samples. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix, high pressure steamed three hours attenuated was better. Different processing methods have different effects on chemical constituents of Polygoni Multiflori Radix. Comparing with crude sample, the contents of gallic acid, 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside, emodin-8-O-beta glucoside and emodin were decreased in processed products with 3 kinds of different methods. The change trend of 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside content was similar with hepatotoxic potency. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix. Processing methods and time attenuated obvious impact on toxicity. Recommended further research on the attehuated standard control of Polygoni Multiflori Radix concocted.
Biological Assay ; Cell Line ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Fallopia multiflora ; chemistry ; toxicity ; Hepatocytes ; drug effects ; Humans ; Plant Roots ; chemistry ; toxicity

The aim of this study was to investigate the effect of Celastrol on induction of HL-60 cell apoptosis and its possible mechanism. The proliferative activity of HL-60 cells treated with 0.25 - 8.0 μmol/L of Celastrol for 24 - 72 hours was assayed by MTT method, the effects of Celastrol on apoptosis and cell cycle of HL-60 were detected by TUNEL staining and flow cytometry with Annexin V-FITC/PI double labeling, the expression of pAkt and cyclin D1 at protein and gene level in HL-60 cells treated with Celastrol were measured by Western blot and RT-PCR. The results showed that the Celastrol could obviously inhibit the proliferation of HL-60 cells in concentration-and time-dependent manners, the IC₅₀ value of Celastrol for 24 hours was 6.21 ± 0.242 μmol/L. The Celastrol concentration-dependently induced the apoptosis of HL-60 cells, accompanying with morphological changes of apoptotic cells, which may be related with arrest of cells in G₀/G₁ phase. The Celastrol suppressed the expression of pAkt and Cyclin D1 in HL-60 cells to a varying degree which showed obvious concentration-and time-dependent manners. It is concluded that the Celastrol inhibits the proliferation and induced the apoptosis of HL-60 cells. Its mechanism may be related with down-regulation of p-Act and cyclin D1 expressions.
Apoptosis ; drug effects ; Cyclin D1 ; metabolism ; Down-Regulation ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-akt ; metabolism ; Triterpenes ; pharmacology

BACKGROUNDSimultaneous pancreas-kidney transplantation (SPKT) frees the diabetic patient with end-stage nephropathy from dialysis and daily insulin injections. Herein, we review consecutive cases of SPKT with bladder drainage performed at our institution over an 8-year period.

METHODSThe study population included 21 patients (16 males and 5 females) who underwent SPKT between September 2001 and September 2009. Seven patients had type-1 diabetes and 14 had type-2 diabetes. Nineteen patients were on dialysis at the time of transplantation. Donation after cardiac death donors were selected for SPKT. The mean human leukocyte antigen match was 2 (range 0 - 4). SPKT was always performed using bladder drainage and vascular anastomoses to the systemic circulation. Immunosuppressive treatment consisted of anti-lymphocyte globulin induction followed by tacrolimus, mycophenolate mofetil, and prednisone.

RESULTSThe mean hospital stay was 45.43 days. After a mean follow-up of 39.4 months, survival rates for patient, kidney, and pancreas were 76.2%, 76.2%, and 66.7% at 1 year; 76.2%, 59.3%, and 55.6% at 5 years; and 57.1%, 39.5%, and 41.7% at 8 years, respectively. Major complications included anastomotic leaks, reflux pancreatitis, and rejection. Six patients died from septic shock (n = 3), duodenal stump leak (1), cardiac arrest (1), or renal failure (1). Eight kidney grafts were lost due to acute rejection (n = 2), chronic rejection (3), and death with a functioning graft (3). Pancreatic graft failure (9) was caused by thrombosis (n = 1), rejection (2), duodenal stump leak (1), and death with a functioning graft (5).

CONCLUSIONSSPKT is a valid therapeutic option for uremic diabetics although few hospitals in China can undertake SPKT.

Adult ; Diabetes Mellitus, Type 1 ; surgery ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Graft Rejection ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Male ; Middle Aged ; Pancreas Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome ; Urinary Catheterization

OBJECTIVEMultivariate analysis was used to select the risk factors in non-insulin dependent diabetes mellitus (NIDDM) patients with oral candidosis, and to establish the forecasting equation, aimed to detect the risk of oral candidosis among NIDDM patients.

METHODS140 NIDDM patients were included in this study. 11 clinical parameters including gender, age, course smoking, fasting blood glucose, oral hygiene status, systemic manifestation, oral mucous membrane status, and denture were recorded respectively. Oral rinse technique was used to detect the salivary candidal carriage. The isolates were identified using CHROM agar Candida test. The Logistic multivariate regression analysis was carried our for risk factors analysis.

RESULTSCandida was found in 69 out of 140 NIDDM cases, and Candida albicans was the major species isolated. The poor glycemic control, poor oral hygiene, and dry mouth were the risk factors of oral candidosis in NIDDM patients, and the forecasting equation was established. Using substitution method, the veracity of the forecasting equation was 82.1%.

CONCLUSIONPoor glycemic control, poor oral hygiene and dry mouth were risk factors of oral candidosis among NIDDM patients. The probability obtained from the forecasting equation may offer references for predicting and preventing the oral candidosis in NIDDM patients.

Adult ; Candida ; Candida albicans ; Candidiasis, Oral ; Diabetes Mellitus, Type 1 ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Oral Hygiene ; Risk Factors ; Smoking

OBJECTIVE Gastric cancer is one of the most common malignant tumors,and the inci-dence rate is the highest in all kinds of tumors in China. However,it remains unclear that how signifi-cantly gastric cells are dependent on glycolysis,and which type of gastric cells are sensitive to glycolysis inhibition. In this study, several kind of gastric cancer cell lines were used as the research object, and the metabolic characteristics of different cell lines were systematically analyzed to provide theoretical support for the accurate treatment of gastric cancer. METHODS We examined the energy metabolism of four gastric cancer cell lines(MGC-803,SGC-7901,HGC-27 and BGC-823)by using glycolysis inhibitor, 2-deoxy-D-glucose(2-DG)and inhibitor of oxidative phosphorylation,oligomycin.Oxygen consumption rates(OCR)and L-lactate were also measured with an XF96 Analyzer(Seahorse Biosciences)to deter-mine the significance of metabolism of oxidative phosphorylation and aerobic glycolysisin gastric cells. In addition, western blot was used to detect the contribution of AMP-activated protein kinase (AMPK), and anti-apoptotic proteins(Bcl-2 and survivin)to clarify the mechanism of death or survival of gastric cancer cells treated by 2-DG or oligomycin. RESULTS In this study, it was shown that the growth of gastric cell lines were suppressed by 2-DG.However,the sensitivity to 2-DG was quite different among cell lines:IC 50 of 2-DG was from 3.28 mmol·L-1(MGC-803)to 15.57 mmol·L-1(BGC-823).MGC-803 was relatively sensitive to 2-DG (IC 50:3.28 mmol·L-1), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, MGC-803 could be more dependent on glycolysis than other cell lines, which was further confirmed by the fact that glucose (+)FCS(-)medium showed more growth and survival than glucose(-)FCS(+)medium.Alternatively, BGC-823, most resistant to 2-DG (IC50: 15.57 mmol·L- 1), was most sensitive to oligomycin, and showed more growth and survival in glucose(-)FCS(+)medium than in glucose(+)FCS(-)medium. Thus,we had reasons to think BGC-823 cells depended on oxidative phosphorylation for energy production. In BGC-823,AMPK,which is activated when ATP becomes limiting,was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented,which might result in resistance to 2-DG.Interestingly,AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in MGC-803,which is 2-DG sensitive. CONCLUSION There is a large metabolic difference between gastric cancer cell lines,which will facilitate the future gastric cancer therapy by targeting metabolic pathways.

OBJECTIVE To probe into the anti-esophagus cancer activity and mechanisms of DN3, a novel natural diterpenoid derivative. METHODS The anti-tumor activity in vitro of DN3 was evaluated by MTT, and by using human esophageal carcinoma cells xenografted into athymic mice model in vivo. The specific mechanisms of DN3, as a dual inhibitor of glycolysis and oxidative phos-phorylation(OXPHOS)were explored through cell and molecular biology techniques.For instance,the manner of cancer cell death induced by DN3 was characterized by hoechst33342, FITC-Annexin V/PI staining and flow cytometric analysis,then these changes of glucose consumption,glucose uptake and lactate production in glycolysis, as well as oxygen consumption rate (OCR) and ATP content in OXPHOS caused by DN3 were performed separately through related kits and SeahorseBioscience XF24 Extra-cellular Flux Analyzer.Furthermore,in order to obtain a clear understanding of the inhibition of DN3 to glycolysis and OXPHOS, these regulatory factors were investigated by Western blot, such as PI3K/AKT, c-Myc and p53 of glycolysis, Bax and HK2 of mitochondrial function. RESULTS DN3 inhibited the growth of esophagus cancer cell EC9706, EC109 and EC1 cells in a dose and time dependent manner,but showed no significant effects on human esophageal epithelial cells(HEECs).DN3 caused significant G2/M arrest of esophagus cancer cell lines and induced apoptosis of these cell lines, which indicated DN3 inhibited the growth of esophagus cancer cell through blocking cell cycle and inducing apoptosis in a dose and time-dependent manner. Importantly, 8 μM DN3 decreased the extracellular acidification rate (ECAR) by 45% in EC109, which indicated glycolysis was inhibited by DN3. Mean-while, DN3 decreased the oxygen consumption rate (OCR) and the OCR linked to intracellular ATP production in EC109 cells,but that was not obvious in HEECs,so which indicated that DN3 could selec-tively block OXPHOS of cancer cells. In addition, the accumulation of reactive oxygen species (ROS) and the drop of mitochondrial membrane potential (MMP) were also observed in EC109 incubated by DN3,which suggested mitochondrial biological function was disturbed.Furthermore,the expression of PI3K/AKT, c-Myc and HK2 related to glycolysis were down-regulated by DN3, but the p53 and Bax were up-regulated in esophageal carcinoma cells. The changes of these enzymes accounted for the decreased glycolysisand OXPHOS in esophageal carcinoma cells treated by DN3. CONCLUSION The new compound DN3 has a strong anti-esophageal carcinoma activity,and it is tolerable that DN3 is seen as a dual inhibitor of glycolysis and oxidative phosphorylation.