Objective To determine the content of Sinapine thiocyanate in the Baixi Kechuan Cream by HPLC and to lay foundation for establishing the quality control standard of the preparation. Methods The HPLC system consisted of the column of DIKMA inertsil (pH=3,5 ?m,250 mm?4.6 mm). The acetonitrile and 3% acetic acid were used as the mobile phase A and B,respectively,and gradient elution was used. The detective wavelength was 326 nm and the flow rate was 1.0 mL/min. The column temperature was room temperature. Results The method was linear within the range of 0.8～4.0 ?g (r=0.999 6,n=5). The average recovery was 100.83%,and RSD=1.95% (n=5). Conclusion The method is quick,simple,highly reproductive and steady,and it can be used for quality control of the preparation.

Objective:To explore the application of EMG and bio-feedback instrument in the recovery of neurological function and hemorheology for patients with stroke.Methods: 65 patients with stroke were enrolled in this study and were randomly divided into combined group (33 cases) and control group (32 cases) as random number table. The patients of combined group were treated by rehabilitation training combined with reconstruction and treatment instrument of neurological function on the basis of basic medicine therapy. The patients of control group were treated only by rehabilitation training on the basis of basic medicine therapy. The clinical effects, the recovery situation of neurological function and the change situation of hemorheology before and after treatment were compared and analyzed.Results: The effective rate of clinical treatment was 96.97% in the combined group and it was 59.38% in the control group, and the difference of effective rate between the two groups was significant (x2=12.473,P<0.05). After treatment, the differences of the daily life behavior Barthel index scale and Fug I -Meyer Assessment (FMA) of motor function score between the two groups were significant (t=7.632,t=5.693;P<0.05). Besides, the plasma viscosity, erythrocyte ratio, platelet aggregation rate, erythrocyte deformation index, erythrocyte aggregation index and fibrinogen of combined group were significantly better than control group, and the differences of them were statistically significant (t=6.859, t=10.263,t=7.626,t=6.623,t=8.257,t=6.003;P<0.05).Conclusion:The reconstruction and treatment instrument of neurological function make a synergistic effect in the treatment of stroke, and it can effectively treat patient with stroke and improve their clinical symptoms. And it also can enhance the motor function of limbs and the self -help ability, and it can significantly improve each indexes of hemorheology of patients. Therefore, it can achieve the better clinical therapeutic effect.

According to the sequence of SmLEA gene,a DNA fragment of 1 038 bp upstream of the coding sequence of SmLEA gene was amplified by DNA walking with the genomic DNA of Salvia miltiorrhiz as the template.Sequence analysis showed that the fragment contained some putative cis-elements relating to abiotic stress,ABA,seed specific expression.So the S.miltiorrhiz seedings was treated with 100?mol/L ABA,200mmol/L NaCl,4℃,and subjected to dehydration.The real-time PCR showed that expression levels of SmLEA was increased obviously,which was in accordance with the sequence analysis.

OBJECTIVETo observe the effect and mechanism of curcumin derivative B06 on kidney from rats with hyperlipidemia and type 2 diabetes.

METHODSThirty five male SD rats were randomly divided into five groups(n = 7): the normal control group, high-fat group, high-fat + B06-treatd group, diabetic group, diabetic + B06-treated group. After fed with high-fat diet for 4 weeks, the later two groups were in- jected with streptozotocin intraperitoneally to induce type 2 diabetes mellitus. B06-treated groups were given B06 by gavage at a dosage of 0.2 mg/kg . d for 8 weeks. After the treatment, the serum creatinine, blood urea nitrogen and uric acid were detected biochemically, the morphology of kidney was observed with light and transmission electron microscopy, the expression of collagen fibers was observed with Masson staining, the protein expression of collogen IV and fibronectin in kidney were determined by Immunohistochemistry.

RESULTSIt was showed that the levels of the serum creatinine and blood urea nitrogen elevated significantly in diabetic group. In high-fat and diabetic groups, increased glomerular mesangial matrix and collagen fiber and thicken glomerular basal membrane were observed under light microscopy, swelling and fusion of foot process were found under electron microscope; increased green matrix within glomeruli was observed under Masson staining. collogen IV and fibronectin protein expression were significantly enhanced in high-fat group and diabetic group. After B06's intervention, the levels of serum creatinine and blood urea nitrogen were decreased in diabetic groups, the morphological change of kidney was obviously relieved, Collogen IV and fibronectin protein expression reduced.

CONCLUSIONCurcumin derivative B06 exerts a protective effect on kidney in type 2 diabetic rats, reduced expressions of collogen IV and fibronectin, inhibition of the accumulation of extracellular matrix and glomerular mesangial proliferation, and then prevention of renal fibrosis may be the mechanism.

Animals ; Blood Urea Nitrogen ; Collagen Type IV ; metabolism ; Creatinine ; blood ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; complications ; drug therapy ; Diabetes Mellitus, Type 2 ; Drugs, Chinese Herbal ; Fibronectins ; metabolism ; Kidney ; metabolism ; physiopathology ; Kidney Diseases ; drug therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Uric Acid ; blood

OBJECTIVETo explore if the antileukemic drugs Vp16 or Ara-C are able to upregulate DR5 gene expression and enhance Apo2L-induced apoptosis of HL-60 cells.

METHODSCell apoptosis was determined by flow cytometry after annexin V/PI staining, the effect of Apo2L on fresh leukemia cells by MTT reduction assay, the expression of DR5 gene in HL-60 cells by semi-quantitative RT-PCR.

RESULTS1. Apo2L induced apoptosis of HL-60 cells in a dose-dependent manner. 2. Apo2L inhibited the proliferation of fresh leukemia cells, but there was difference among different subtypes. 3. Vp16 or Ara-C upregulated DR5 gene expression and augmented Apo2L-induced apoptosis in HL-60 cells.

CONCLUSIONApo2L could induce apoptosis of HL-60 cells and inhibit the proliferation of fresh leukemia cells. Ara-C or Vp16 upregulated DR5 gene expression and increased the sensitivity of HL-60 to Apo2L-induced cytotoxicity. Apo2L might be a promising antileukemic agent for the treatment of leukemia.

Antineoplastic Agents ; pharmacology ; Apoptosis Regulatory Proteins ; Cytarabine ; pharmacology ; Drug Synergism ; Etoposide ; pharmacology ; Female ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; Male ; Membrane Glycoproteins ; pharmacology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Necrosis Factor-alpha ; pharmacology

AIM:To observe the changes in heme oxygenase-1(HO-1) expression in the lung after ischmia-reperfusion of hind limbs in rats.METHODS:Hind limbs ischemia was made by clamping infrarenal aorta with a microvascular clip and lung injury was made by following reperfusion. Lung tissue was obtained from the animals subjected to sham operation, 4 h ischemia without reperfusion and 4 h, 8 h, 16 h, 24 h, 48 h reperfusion following 4 h ischemia. The levels of HO-1 mRNA and protein were measured at different times by Northern blot and Western blot. Immunohistochemistry technique was used to determine the cell types responsible for limb ischemic reperfusion induced HO-1 expression. RESULTS:After ischemia-reperfusion of limbs, HO-1 mRNA increased by 4 h, reached a peak at 16 h, and returned toward baseline at 24-48 h. This time course correlated with increased HO-1 protein. Immunohistochemical studies showed HO-1expressed in a variety of cell types, including the airway epithelium, alveolar macrophages and vascular smooth muscular cells. There were no positive signals in sham group and ischemia group both in mRNA levels and protein levels. CONCLUSION:The expression of HO-1 in the lung is not induced by limb ischemia or sham operation, but induced by limb reperfusion after ischemia in rats.

AIM: To observe the changes in heme oxygenase-1(HO-1) expression in the lung after ischmia-reperfusion of hind limbs in rats.METHODS: Hind limbs ischemia was made by clamping infrarenal aorta with a microvascular clip and lung injury was made by following reperfusion. Lung tissue was obtained from the animals subjected to sham operation, 4 h ischemia without reperfusion and 4 h, 8 h, 16 h, 24 h, 48 h reperfusion following 4 h ischemia. The levels of HO-1 mRNA and protein were measured at different times by Northern blot and Western blot. Immunohistochemistry technique was used to determine the cell types responsible for limb ischemic reperfusion induced HO-1 expression. RESULTS: After ischemia-reperfusion of limbs, HO-1 mRNA increased by 4 h, reached a peak at 16 h, and returned toward baseline at 24-48 h. This time course correlated with increased HO-1 protein. Immunohistochemical studies showed HO-1expressed in a variety of cell types, including the airway epithelium, alveolar macrophages and vascular smooth muscular cells. There were no positive signals in sham group and ischemia group both in mRNA levels and protein levels. CONCLUSION: The expression of HO-1 in the lung is not induced by limb ischemia or sham operation, but induced by limb reperfusion after ischemia in rats.

OBJECTIVETo investigate the effect of Tongshenluo Capsule (TSL) on the components of extracellular matrix (ECM) and their metabolism in kidney of rats with diabetic nephropathy (DN), and to explore its mechanism of kidney protecting.

METHODSThe DN model rats established by one side nephrectomy and intraperitoneal injection of streptozotocin were randomly divided into 5 groups, the sham-operation group, the model group, the Valsartan group, the Gliquidon group, and the TSL group, 10 in each group. Levels of fasting blood glucose (FBG) and 24-h urinary micro-content of albumin (24 h mAlb) were determined dynamically; the serum glycosyl hemoglobin (HbA1c)was determined after the last medication; the ultrastructural changes of kidney were observed by transmission electron microscope; the expressions of collagen IV (IV-C), fibronctin (FN), laminin(LN), and the ECM metabolism influencing factors, including MMP-2, tissue inhibitor of metalloproteinase (TIMP-2), transfer growth factor-beta1 (TGF-beta1) in renal tissue were detected by immunohistological chemistry and image collecting analytical system. Results TSL could decrease the levels of FBG, HbA1c, 24 h mAlb (P < 0.05 or P < 0.01), ameliorate the thickness of glomerular basement membrane (GBM), decrease the components of ECM, down-regulate TGF-beta1 and TIMP-2 expression, and up-regulate MMP-2 expression (P < 0.05 or P < 0.01).

CONCLUSIONTSL plays a role of kidney protection by decreasing the ECM components expression and regulate ECM metabolism.

Animals ; Capsules ; Collagen Type IV ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Diabetic Nephropathies ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Laminin ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.

Objective To evaluate the performance of HbA1C in diagnosis of pre-diabetes in Chinese adolescents.Methods A total of 933 students(aged 13-16 year,46.5% female)without known history of diabetes were selected from junior and senior middle schools in Liaoyang city.HbA1C and fasting plasma glucose(FPG)levels were determined in all participants.Pre-diabetes was diagnosed as according to 2010 American Diabetes Association (ADA)criteria[HbA1C 5.7%-6.4%,FPG(5.6-6.9)mmol/L].The individuals with HbA1C ≥5.7% underwent oral glucose tolerance test(OGTT)and were classified into impaired glucose tolerance(IGT)and impaired fasting glucose(IFG)according to 1999 World Health Organization(WHO)criteria.The performance of HbA1C in diagnosing pre-diabetes by the new criteria was evaluated by the traditional OGTT results based on receiver operating characteristic(ROC)curves.Results Among all participants,213(22.8%)individuals were diagnosed as prediabetic subjects.72.3% of individuals with pre-diabetes were identified by HbA1C alone,23.0% by FPG alone,and only 4.7% by both tests simultaneously.The ROC curve for HbA1C to identify impaired fasting glucose diagnosed by new criteria had an area under the curve(AUC)of 0.50 without statistical significance(P = 0.97).Of the 164 participants with HbA1C 5.7% -6.4%,68 individuals received OGTT,among which 18 subjects were diagnosed as IGT(n= 14)or IFG(n = 4).The ROC curve for HbA1C to identify pre-diabetes diagnosed by OGTT had an AUC of 0.53,without statistical significance(P = 0.69).Conclusion In Chinese adolescents,HbA1C is not suitable for diagnosis or screening of pre-diabetes.