OBJECTIVETo study the molecular mechanism of extracts from Euodia rutaecarpa on hepatotoxicity in mice.

METHODTotally 30 KM mice were divided into 3 groups and orally administrated extracts from E. rutaecarpa for consecutively 15 days. The expressions of Erkl/2, CDK8, CK1e, Stat3 and Src were detected by Western blotting method.

RESULTThe extracts from E. rutaecarpa could up-regulated Erkl/2, CDK8 and CK1e expressions (P <0.01) and down-regulate Stat3 and Src (P <0.01).

CONCLUSIONThe molecular mechanism of E. rutaecarpa on hepatotoxicity may be correlated with Erkl/2, CDK8, CKle, Stat3 and Src signal molecules.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Down-Regulation ; Drugs, Chinese Herbal ; toxicity ; Evodia ; chemistry ; Female ; Fruit ; chemistry ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; metabolism ; Male ; Mice ; Plants, Medicinal ; Signal Transduction ; drug effects ; Specific Pathogen-Free Organisms ; Triglycerides ; blood ; Up-Regulation

Objective To evaluate if continuous hemihepatic inflow occlusion(HH)during hepatectomy can be as safe and effective as intermittent total hepatic inflow occlusion(TH)in reducing blood loss during hepatectomy.Methods From November 2001 to March 2006.eighty patients undergoing liver resections were included in a prospective randomized study comparning the intra-and postoperative course underTH(n=40)or HH(n=40).TH was performed with periods of 20 minutes of occlusion and 5 minutes of releasing,while HH with continuous occlusion.The surface area of liver transection was measured and blood loss was calculated.The amount of blood loss,levels of alanine aminotransferuse (ALT)and aspartate aminotransferase(AST),and postoperative course were recorded. Results The total ischemic time of the HH groups was longer than in the TH group[(42±13)min,(31±13)min,P=0.37],and the operative time in the HH group was longer than in the TH group[(236 ±49)min,(204±38)min,P=0.02 ].No signincant difierenee was found between HH and TH group in blood loss during liver parenchyma transection[(500 ±269)ml,(416 ±235)ml,P=0.14]and in the changes of ALT and AST on the first postoperative day[ALT:(677±572)IU/L,(577 ±327)IU/L,P=0.12;AST:(591 ±468)IU/L,(512±301)IU/L,P=0.66].There were no difierences on postoperative morbidity between the two groups(22.5%versus 20.0%,P=0.35).Conclusion The technique of continuous hemihepatic inflow occlusion is as safe and effective as intermittent total hepatic inflow occlusion.

Objective To assess the value of prenatal sonographic diagnosis on fetal agenesis of corpus callosum(ACC). Methods A retrospective study was on prenatal sonographic findings of 10 cases with ACC malformation and their abnormalities in central nervous system (CNS) or extra-CNS.Results The special sonographic findings established the diagnosis of ACC malformation in all 10 cases,with 7cases diagnosed absence agenesis. Among all patients with ACC,6 cases were accompanied with abnormalities in extra-CNS,8 in CNS and 5 in both. Conclusions Prenatal ultrasonography plays a vital role in accurate diagnosis on fetal ACC. Attentions should be paid to the indirect encephalic features and complicated abnormalities so as to make accurate and prompt diagnosis.

OBJECTIVETo study the regulation of luteolin on spleen cells and sarcoma S180 cells in normal ICR mice.

METHODSSpleen cells and S180 cells were incubated with different concentrations of luteolin (50, 100, 200, and 400 μmol/L). The effect of luteolin on spleen cells and sarcoma S180 cells was determined by MTT assay. The apoptosis was detected using propidium iodide staining flow cytometry. Intracellular reactive oxygen species (ROS) was determined by flow cytometric analysis. Activities of free radicals scavenging were determined by hydroxyl radical and DPPH tests.

RESULTSCompared with the solvent control group, 200 and 400 μmol/L luteolin increased the spleen cells viability (P < 0.05). Luteolin at 100, 200, and 400 μmol/L decreased activities of S180 cells (P < 0.01). The proportion of sub-G1 phase spleen cells was reduced after treated with 200 and 400 μmol/L luteolin (P < 0.05). The proportion of sub-G1 phase S180 cells was elevated after treated with 200 and 400 μmol/L luteolin (P < 0.05). Compared with the solvent control group, levels of intracellular ROS in spleen cells of ICR mice all increased; levels of intracellular ROS in S180 cells all decreased after treated with 50, 100, 200, and 400 μmol/L luteolin (P < 0.05). Luteolin scavenged hydroxyl radical and DPPH in a dose dependent manner.

CONCLUSIONLuteolin had bilateral regulation on viability and apoptosis of spleen cells and S180 cells (promoting the viability of spleen cells, inhibiting apoptosis of spleen cells, inhibiting the viability of S180 cells, and promoting apoptosis of S180 cells), which was worth further study and exploration.

Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Survival ; Luteolin ; metabolism ; Mice ; Mice, Inbred ICR ; Reactive Oxygen Species ; Sarcoma ; Spleen ; metabolism

BACKGROUND: Early reperfusion can effectively treat acute myocardial infarction (AMI) and reduce the mortality significantly. This study aimed to compare the role of plasma microRNA-1 (miR-1) and cardiac troponin T (cTnT) in early diagnosis of AMI patients. METHODS: From May 2011 to May 2012, plasma samples were collected from 56 AMI patients and 28 non-AMI controls. The expression of plasma miR-1 was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and the level of plasma cTnT was measured using electrochemiluminescence-based methods on an Elecsys 2010 Immunoassay Analyzer. SPSS 16.0 was used for the statistical analysis of the results. Data were expressed as mean±standard deviation unless otherwise described. The differences about clinical characteristics between the AMI patients and controls were tested using Student'st test or Fisher's exact test. The Mann-WhitneyU test was conducted to compare the expression of microRNAs between the AMI patients and controls. MicroRNAs expression between different intervals of the AMI patients was compared using Wilcoxon's signed-rank test. The receiver operating characteristic (ROC) curve was established to discriminate the AMI patients from the controls. RESULTS: In the present study, the expression of plasma miR-1 was significantly increased in the AMI patients compared with the healthy controls (P<0.01). The plasma miR-1 in the AMI patients decreased to the normal level at 14 days (P>0.05). The expression of plasma miR-1 was not related to the clinical characteristics of the study population (P>0.05). ROC curve analyses demonstrated that miR-1 was specific and sensitive for the early diagnosis of AMI, but not superior to cTnT. CONCLUSION: Plasma miR-1 could be used in the early diagnosis of AMI, but it is similar to cTnT.

OBJECTIVETo observe the effect of recombinant interleukin-6 (IL-6) and osteoprotegerin (OPG) on inhibiting bone absorption induced by receptor activator for nuclear factor-kB ligand (RANKL) in murine osteoclast precursor cells (OCPs) model.

METHODSRAW 264.7 cells were solely treated with 50 ng/ml RANKL for 1 day, and then they were divided into three groups: RANKL (control group), RANKL+IL-6 (IL-6 group) and RANKL+IL-6+OPG (combination group). These cells were harvested and investigated by means of HE staining under light microscope after consecutive 9 days. Furthermore, staining tartrate-resistant acid phosphatase(TRAP)-positive multinucleated cells were detected by inverted phase contrast microscope. The absorption pits of bone slices were observed under scanning electron microscope.

RESULTSThe number of mature osteoclast cells in control group was more than that in IL-6 alone or IL-6 combined with OPG group (P less than 0.05). Interestingly, this experiment has also demonstrated that there was a large number of TRAP-positive multinucleated osteoclasts (more than 3 nuclei) and several bone absorption formation in the control group, whereas the outcome was completely different in both IL-6 group and IL-6+OPG group (P less than 0.05).

CONCLUSIONIL-6 can suppress the differentiation of mature osteoclasts as directly adding it into the RAW 264.7 cells induced by 50 ng/ml RANKL, and further the effect of osteolysis is remarkably reduced. When treatment with IL-6 combined with OPG, a more effective strategy for the treatment of osteoporosis is reached.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Interleukin-6 ; administration & dosage ; pharmacology ; Mice ; Osteoclasts ; cytology ; drug effects ; Osteoprotegerin ; administration & dosage ; pharmacology ; RANK Ligand ; pharmacology ; Recombinant Proteins ; administration & dosage ; pharmacology

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

OBJECTIVETo observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes.

METHODLymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments.

RESULTThe results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent.

CONCLUSIONThe comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.

Animals ; Calorimetry ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Isatis ; chemistry ; Lymphocytes ; drug effects ; Male ; Mice ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.

METHODSHUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.

RESULTSs The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.

CONCLUSIONDigestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.

Cell Culture Techniques ; Cell Cycle ; Cells, Cultured ; Culture Media ; chemistry ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 8 ; chemistry ; Serum

OBJECTIVETo explore the Video-urodynamic characteristics of various neurogenic bladder.

METHODSA total of 1800 patients with neurogenic bladder were included in our study from December 2002 to June 2008. All patients underwent Video-urodynamic studies. Urodynamic data was collected and analyzed.

RESULTSUrodynamic study showed detrusor overactivity in 71%, of which 60% with uninhibited sphincter relaxation, and acontractile detrusor in 29% stroke patients. No upper urinary tract deterioration was found in all 42 stroke patients. Detrusor overactivity without sphincter dyssynergia was found in 70% patients with head trauma. Seven patients with Parkinson disease showed detrusor overactivity, of which 3 with delayed sphincter relaxation. Detrusor overactivity was found in 91% and detrusor sphincter dyssynergia in 83% supra-sacral spinal cord injured patients. Acontractile detrusor was found in 73% patients with conus medullaris and cauda equina injury. Overall, upper urinary tract changes were found in 12% and vesicoureteral reflux in 4% spinal cord injured patients. Urodynamic study showed acontractile detrusor in 81%, reduced compliance in 86%, upper urinary tract changes in 55% and vesicoureteral reflux in 33% patients with myelodysplasia. Most patients (92%) with protruded lumbar disc showed detrusor areflexia. Normal bladder compliance was found in 88% patients with protruded lumbar disc. Urodynamic study showed reduced bladder sensation in 81% and detrusor under-activity in 76% patients with diabetic urinary bladder disease.

CONCLUSIONSVideo-urodynamic study can provide the most detailed information about the bladder dysfunction. It is the most valuable examination before treatment of patients with neurogenic bladder.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Urinary Bladder, Neurogenic ; diagnosis ; etiology ; physiopathology ; Urodynamics