Sixteen compounds have been isolated from the EtOAc fraction of 95% ethanolic extract of Sophora dunnii through silica gel, Sephadex LH-20 and semi-prerarative HPLC column chromatographies. Their structures were identified on the basis of NMR and MS spectra data as phaseollidin (1), L-maackiain (2), 2-(2',4'-dihidroxyphenyl)-5,6-methylenedioxy benzofuran (3), 8-demethyl-farrerol (4), liquiritigenin (5), genistein (6), 6-methylgenistein (7), 5-O-methyl genistein (8), 7,2',4'-trihydroxys-5-methoxy-isoflavanone (9), 7, 3', 4'-trihydroxy-isoflavanone (10), erythribyssin D (11), calycosin (12), trans-resveratrol (13), cis-resveratrol (14), stigmasterol (15), β-sitosterol (16). Among these, compounds 1-14 and 16 were isolated from this plant for the first time.
Chemical Fractionation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Sophora ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Adult ; China ; epidemiology ; GB virus C ; classification ; Hepatitis, Viral, Human ; epidemiology ; virology ; Homosexuality, Male ; Humans ; Male

Antineoplastic Agents ; Humans ; Leukemia, Myeloid, Acute/drug therapy* ; Mutation ; Protein Kinase Inhibitors/therapeutic use* ; fms-Like Tyrosine Kinase 3

OBJECTIVETo investigate the effects of protein tyrosine kinase (PTK), protein tyrosine phosphatase (PTP) and protein kinase C (PKC) on apoptosis and observe the changes of cytosolic calcium ([Ca(2+)]i) of arsenic trioxide (As2O3) treated human leukemia cells NB4 and cortex neurons.

METHODS[Ca(2+)]i of NB4 cells and cortex neurons was probed with Fluo-3/AM, its changes were assayed with laser confocal microscopy in real-time after As2O3 treatment at different concentrations, the effects of PTK and PTP and the activation of PKC on these changes with confocal microscopy and phosphorus radioisotope assay. DNA ladders of NB4 cells and cortex neurons after exposed to As2O3 were observed.

RESULTSAs2O3 at 1 micromol/L could remarkably increase the [Ca(2+)]i of NB4 cells but had no effects on neurons. Vanadate, a kind of PTP inhibitor, could promote the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total increase rates at 240 seconds after exposed to As2O3 at different concentrations were (6.5 +/- 2.3)%, (21.7 +/- 2.1)%, (49.2 +/- 2.5)% for NB4 cells, and (6.7 +/- 2.1)%, (19.4 +/- 2.5)%, (52.3 +/- 2.7)% for cortex neurons, respectively. Genistein, a kind of PTK inhibitor, could decrease the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total inhibited rates at 240 seconds after As2O3 treatment at different concentrations were (6.7 +/- 2.9)%, (25.6 +/- 2.5)%, (52.2 +/- 3.5)% for NB4 cells, and (7.8 +/- 3.1)%, (18.1 +/- 2.8)%, (51.3 +/- 3.3)% for cortex neurons, respectively. The activation of PKC began to increase as exposed to As2O3 at 1 micromol/L for 3 h, and kept rising continuously in NB4 cells and at 24 h DNA ladders emerged. However, none of the above results was found in human cortex neurons, but when exposed to 2 micromol/L As2O3, the activation of PKC and DNA ladders did emerge in neurons.

CONCLUSIONSThe phosphorylation and dephosphorylation of PTK and PTP participated in nonspecific apoptosis signal transduction pathway related to As2O3, and accompanied with PKC activation. The [Ca(2+)]i elevation was closely related to increased PKC activation. There existed difference in dose tolerances to As2O3 between NB4 cell and cortex neurons.

Adult ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Calcium ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Cerebral Cortex ; cytology ; Cytoplasm ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Leukemia, Promyelocytic, Acute ; enzymology ; metabolism ; pathology ; Male ; Neurons ; cytology ; drug effects ; metabolism ; Oxides ; pharmacology ; Protein Kinase C ; metabolism ; Protein Tyrosine Phosphatases ; metabolism ; Protein-Tyrosine Kinases ; metabolism

OBJECTIVETo explore the alterations of apoptosis factor Bcl-2/Bax in the early Parkinson's disease (PD) rats and the protective effect of scorpion venom derived bioactive peptide.

METHODSHealthy male SD rats (180-220 g) were randomly divided into 4 groups (n = 10): early PD model group, sham operation group, scorpion venom derived bioactive peptide control group, scorpion venom derived bioactive peptide therapy group. 6-hydroxydopamine (6-OHDA) was used to prepare the early PD rat model. The immunohistochemistry was used to detect the expression of Bax and Bcl-2 and further explore the mechanism of anti-apoptosis regarding the neuroprotective effect of scorpion venom derived bioactive peptide.

RESULTSThe results indicated that compared with the control rats, the immunostaining of Bax in the brain increased significantly while that of Bcl-2 decreased significantly in the lesion side of 6-OHDA treated rats. Interestingly, scorpion venom derived bioactive peptide could attenuate the above abnormal changes.

CONCLUSIONUp-regulation of Bax and down-regulation of Bcl-2 could participate in the early stage of PD and the anti-apoptotic mechanism could be involved in the neuroprotective effect exerted by scorpion venom derived activity peptide regarding the dopaminergic neuron in the early stage.

Animals ; Apoptosis ; Disease Models, Animal ; Down-Regulation ; Male ; Neuroprotective Agents ; chemistry ; Oxidopamine ; Parkinson Disease ; metabolism ; Peptides ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; chemistry ; Up-Regulation ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the changes in the PGE(2) and PGI(2), TXA(2) levels and PGT mRNA expression in the intestinal mucosa in scalded rats.

METHODSWistar rats inflicted with TBSA 30% III degree scald were employed as the model. The PGE(2) and PGI(2) and TXA(2) contents in the intestinal mucosa were measured by radioimmunoassay, and the expression of PGT mRNA was detected by in situ hybridization.

RESULTSThe PGE(2) and PGI(2) levels in intestinal mucosa were increased at 12 postburn hours (PBHs) and thereafter decreased dramatically (P < 0.05). The TXA(2) level in intestinal mucosa of scalded rats was obviously higher than that of normal level at 24 and 48 PBHs (P < 0.05), and the expression of PGT mRNA seemed to be increased after scalding.

CONCLUSIONThe decrease of PGE(2) level and the increase of TXA(2) level in the intestinal mucosa of scalded rats might be involved in rat mucosal injury, and PGT played an important role in the regulation of PGs levels.

Animals ; Burns ; metabolism ; pathology ; Intestinal Mucosa ; metabolism ; pathology ; Organic Anion Transporters ; metabolism ; Prostaglandins ; analysis ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo explore the different effects of systolic blood pressure (SBP) and low density lipoprotein on carotid plaques (LDL-C).

METHODSA total of 101 510 serving and retired workers of a company who participated in the health examination in 2006-2009, 5852 participants were selected as study subjects by stratified random sampling according to the age and sex ratio. These subjects took their health examination in 2010-2011 including the carotid ultrasound. Finally, 5361 eligible participants with complete data were included in the analysis. The detection and weighted rates of carotid plaques were calculated for four groups: normal SBP and LDL-C group (3524 subjects), normal SBP and high LDL-C group (356 subjects), elevated SBP and normal LDL-C group (1308 subjects) and elevated SBP and high LDL-C group (173 subjects). The effects of different baseline SBP and LDL-C on detection rates of the carotid artery plaques were analyzed by logistic regression.

RESULTSThe detection rate of carotid plaques in normal SBP and LDL-C group, normal SBP and high LDL-C group, elevated SBP and normal LDL-C group, elevated SBP and high LDL-C group was 33.7% (1186/3524), 41.3% (147/356), 64.8% (847/1308), 68.8% (119/173) (χ(2) = 425.75, P < 0.05) and the weighted detection rate was 36.0%, 42.0%, 64.5% and 68.3% respectively. For men, the detection rate was 44.2% (877/1985), 51.1% (97/190), 70.6% (657/930), 71.3% (82/115) (χ(2) = 194.02, P < 0.05) and the weighted detection rate was 31.2%, 36.1%, 49.8% and 50.3% respectively. For women, the detection rate was 20.1% (309/1539), 30.1% (50/166), 50.3% (190/378), 63.8% (37/58) (χ(2) = 180.17, P < 0.05) and the weighted detection rate was 30.9%, 46.3%, 70.3%, and 88.1% respectively. After adjusted for other risk factors, the OR (95%CI) value was 1.37 (1.05 - 1.78), 2.05 (1.74 - 2.43) and 2.12 (1.45 - 3.12) for normal SBP and high LDL-C group, elevated SBP and normal LDL-C group and elevated SBP and high LDL-C group respectively compared with normal SBP and LDL-C group.

CONCLUSIONElevated SBP and high LDL-C were risk factors of the carotid artery plaques. Compared with high LDL-C, elevated SBP may add a higher risk for carotid plaques.

Adult ; Aged ; Blood Pressure ; Carotid Stenosis ; blood ; epidemiology ; physiopathology ; Cholesterol, LDL ; blood ; Dyslipidemias ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Systole

OBJECTIVETo study the clinical application value of middle segment pancreatectomy in the treatment of benign tumors of the amphi-neck of the pancreas.

METHODSFifteen cases were retrospectively analyzed treated from November 2005 to June 2009. There were 3 male and 12 female aging from 30 to 50 years. They all received middle segment pancreatectomy for benign tumors of the amphi-neck of the pancreas.

RESULTSThere was no death during perioperative period. All the 15 patients received middle segment pancreatectomy. Fourteen of them received the closure of broken ends of pancreatic head, pancreaticojejunostomy (mono-anastomosis) and the rest one received dipl-anastomosis. Postoperative pathology showed that in the 15 patients, 1 got solid-pseudopapillary tumor of the pancreas, 3 got non-functional islet cell tumor, 11 got cystadenoma of pancreas. Three of them got pancreatic fistula and were self cured in 3 months. Follow-up visits to all the patients kept in the following 2 to 45 months. There was no death. No patients got new-onset diabetes and pancreatic pseudocyst. And their tumors were not relapsed.

CONCLUSIONSThere is an exact therapeutic effect of middle segment pancreatectomy for benign tumors of the amphi-neck of the pancreas. The treatment has little function damage to patients' endocrine and external secretion. The incidence rate of pancreatic fistula in middle segment pancreatectomy is higher than that in pancreaticoduodenectomy. As long as the drainage is kept unobstructed, most of the pancreatic fistula can be self cured.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; surgery ; Retrospective Studies

OBJECTIVETo develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system.

METHODSThe heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector Pac-kappa-Fc and Pac-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized.

RESULTSThe baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16.

CONCLUSIONThe recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection.

Antibodies, Monoclonal ; biosynthesis ; immunology ; Baculoviridae ; genetics ; Hepatitis A virus ; immunology ; Hepatitis Antibodies ; biosynthesis ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; immunology ; Immunoglobulin G ; biosynthesis ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Recombinant Proteins ; biosynthesis ; immunology

OBJECTIVETo explore the way to induce mesenchymal stem cells (MSCs) to differentiate into dopaminergic neurons in vitro.

METHODSMSCs were obtained from rat bone marrow, cultured and passaged. MSCs used in this experiment had multipotency, which was indirectly proved by being induced to differentiate into chondrocytes and adipocytes. MSCs were cultured in medium containing 0.5 mmol/L IBMX for 2 days. Then the medium was replaced with induction medium, which contained GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments. The surface markers of the differentiated neurons, such as NSE, nestin, MAP-2a, b and TH were detected by immunocytochemistry and Western blot after MSCs were cultured in induction medium for 7 days and 15 days.

RESULTSMSCs differentiated into neural progenitors and expressed nestin after MSCs were incubated with medium containing IBMX for 2 d. After the medium was replaced with induction medium containing many inducing agents, MSCs differentiated into neuron-like cells and dopaminergic neuron-like cells and expressed NSE, MAP-2a, b and TH. The percentage of NSE-positive cells, MAP-2a, b-positive cells and TH-positive cells was 30.032 +/- 2.489%, 41.580 +/- 5.101% and 34.958 +/- 5.534%, respectively after MSCs were induced in medium containing GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments for 15 days.

CONCLUSIONMSCs can differentiate into dopaminergic neuron-like cells and are a new cell source for the treatment of neurodegeneration diseases and have a great potential for wide application.

Adipocytes ; cytology ; Animals ; Blotting, Western ; Bone Marrow Cells ; Carboxylesterase ; analysis ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Culture Media, Conditioned ; Dopamine ; analysis ; Intermediate Filament Proteins ; analysis ; Mesencephalon ; cytology ; Mesenchymal Stromal Cells ; cytology ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; cytology ; metabolism ; Phosphoprotein Phosphatases ; analysis ; Rats ; Rats, Wistar