Objective To explore the influence of homocysteine(Hcy)on liver cystathionine-?-synthase(CBS)and methylenetetrahydrofolate reductase(MTHFR)system in apoE-/- mice,and determine the effects of atorvastatin and/or folate/vitamin B12 on liver CBS and MTHFR system.Methods Eighty male 6-week-old apoE-/- mice were randomly divided into two groups:65 mice were fed with a chow diet containing 2%(wt/vol)L-methionine(homomethionine group)and 15 mice were fed with normal saline(control group).Two months later,the 60 mice survived in homomethionine group were subdivided into four groups:group Ⅰ(untreated),Ⅱ(3 mg/kg atorvastatin),Ⅲ(3 mg/kg atorvastatin+2 mg/kg folate+30 ?g/kg vitamin B12)and Ⅳ(2 mg/kg folate+30 ?g/kg vitamin B12).After one month,Western blotting was performed to detect the liver CBS and MTHFR system protein expression in each group.Results The relative expression of liver CBS and MTHFR was significantly lower in group Ⅰ than in control group(P

The contents of coumarins in the sulfur fumigated Angelicae Dahuricae Radix (Baizhi, ADR) were reduced significantly. To achieve the quality control of ADR, the qualitative identification of sulfur fumigated ADR and quantitative model of imperatorin content should be established. The near-infrared (NIR) spectrograms of non-sulfur and sulfur fumigated ADR were collected by NIR diffuse reflectance spectroscopy technology and pretreated by the method of first derivative derivation and vector normalization. The Ward's Algorithm method was used for the cluster analysis. The non-sulfur and sulfur fumigated ADR can be quickly identified in the range of 8,806. 0-3 811.0 cm(-1) based on the cluster analysis. The NIR quantitative model of imperatorin was established by the contents of imperatorin determined by HPLC in combination with partial least squares regression analysis. According to the calibration model established in this study, correlation coefficients (R2), the root-mean-square error of cross-validation (RMSECV), and the root-mean-square error of prediction (RMSEP) for imperatorin were 0.982 8, 0.006 8, 0.011 8, respectively. The quantitative model of imperatorin can be applied to determine the content of imperatorin in ADR accurately.
Angelica ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Sulfur ; chemistry

Adolescent ; Adult ; Female ; Health Education ; Health Knowledge, Attitudes, Practice ; Health Services Needs and Demand ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Rural Population ; statistics & numerical data ; Surveys and Questionnaires ; Transients and Migrants ; statistics & numerical data ; Young Adult

Objective To investigate whether homocysteine(Hcy) induces apoptosis of endothelial cells via a pathway involving caspases3 and whether simvastatin antagonizes the proapoptotic effects of Hcy by regulating c-IAP. Methods Human umbilical vein endothelial cells(HUVEC) were treated with Hcy,with or without simvastatin,for 24 h.Cell apoptosis was evaluated by Annexin V staining and flow cytometery,as well as TUNEL.The mRNA and protein levels of caspase3,c-IAP-1 and c-IAP-2 were analyzed by RT-PCR and Western blot,respectively. Results Treatment with both low(0.5 mmol/L) and high(3.0 mmol/L) concentrations of Hcy-induced HUVEC apoptosis was accompanied by an increased level of caspase3 expression and activation,together with decreased c-IAP-1 and c-IAP-2 level.Simvastatin upregulated c-IAP-1 and c-IAP-2 expression while attenuated Hcy-induced apoptosis and caspase3 activation. Conclusion Hcy may induce HUVEC apoptosis via a pathway involving caspase3,which can be partially antagonized by simvastatin,possibly through upregulated c-IAP-1 and c-IAP-2 expression.

Anesthetics, Intravenous ; administration & dosage ; pharmacology ; Animals ; Animals, Newborn ; Cerebral Cortex ; cytology ; metabolism ; Drug Synergism ; Female ; Fentanyl ; administration & dosage ; pharmacology ; Male ; Midazolam ; administration & dosage ; pharmacology ; Neurons ; metabolism ; Patch-Clamp Techniques ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Voltage-Gated Sodium Channels ; drug effects

UNLABELLEDOBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test.

RESULTSCompared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05).

CONCLUSIONclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glycation End Products, Advanced ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Stem Cells ; cytology ; drug effects

Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.

OBJECTIVETo investigate the effects of antisense oligonucleotides (ASODN) targeting protein kinase C alpha (PKCalpha) on the proliferation of A549 cells.

METHODSPKCalpha ASODN and random oligonucleotides (RODN) were transfected into A549 cells mediated by polyethyleneimine, and the proliferation and clone formation of A549 cells were detected by CCK-8 and clone formation assay, respectively. The expression of PKCalpha in the transfected cells was analyzed by RT-PCR and Western blotting.

RESULTSCompared with those in the control group, PEI group and PEI-RODN group, the proliferation and clone formation of A549 cells treated with ASODN targeting PKCalpha were significantly inhibited (P<0.05). The expressions of PKCalpha mRNA and protein in PKCalpha ASODN-transfected A549 cells were significantly lower than those in the other 3 groups (P<0.05).

CONCLUSIONThe PKCalpha ASODN mediated by PEI down-regutates the expression of PKCalpha gene and suppress the proliferation and clone formation of A549 cells.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Lung Neoplasms ; enzymology ; pathology ; Oligonucleotides, Antisense ; genetics ; pharmacology ; Protein Kinase C-alpha ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Transfection

AIMTo establish a method for determinate of the inhibitory activity of angiotensin-converting enzyme inhibitor captopril by high performance capillary electrophoresis.

METHODSThe characteristic absorptive wavelength of hippuric acid determined by ultraviolet spectrophotometer is 228 nm. The method employed a melted capillary column, 50 mmol.L-1 phosphoric acid (pH 8.3) buffer solution, inject pressure 4.8 kPa, inject time 3 s, separation voltage 20 kV and detection wavelength 228 nm.

RESULTSThe reactant and resultant was separated completed within 7 min. IC50 of captopril was 0.019 mumol.L-1. Captopril is a competitive inhibitor, which was proved by enzyme reaction dynamics.

CONCLUSIONThe method was shown to be accurate, simple and rapid and can be used for determination of the inhibitory activity of captopril.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Captopril ; pharmacology ; Electrophoresis, Capillary ; methods ; Hippurates ; analysis ; Peptidyl-Dipeptidase A ; metabolism

AIMTo establish quality control requirements and methods for recombinant adeno-associated virus(rAAV) type 2/human blood coagulation factor IX (rAAV-2/hFIX).

METHODSIdentification of rAAV genome fragments, potential contaminants including wild type AAV(wtAAV) and helper virus, were detected by PCR. Purity of rAAV-2/hFIX was analyzed by cation-exchange HPLC and SDS-PAGE. Virus partical numbers were performed by dot blot assay. hFIX expression was demonstrated by ELISA and potency of hFIX was verified by APTT.

RESULTSIdentity of rAAV-2/hFIX was proved. Residues of wtAAV and helper virus were conformed to requirements. Purity of rAAV-2/hFIX were more than 98%. Partical numbers of rAAV-2/hFIX were more than 1.0 x 10(15) VG.L-1. hFIX expression was more than 20.0 micrograms.L-1. hFIX potency was verified by APTT following rAAV-2/hFIX injected to FIX gene knockout mice, potency results conformed to requirements.

CONCLUSIONThe methods and requirements had been established for quality control of rAAV-2/hFIX.

Animals ; Dependovirus ; genetics ; Factor IX ; biosynthesis ; genetics ; Female ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Genome, Viral ; Humans ; Male ; Mice ; Mice, Knockout ; Quality Control ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics