This study was to examine the mechanism of oleanolic acid (OA) induces G2/M phase arrest and apoptosis in human hepatocellular carcinoma Bel-7402 cells. MTT and trypan blue exclusion test assay were adopted to detect the proliferate status of cells treated with OA. We assayed the cell cycle by flow cytometry using PI staining. Apoptosis was determined by Annexin V-FITC staining and PI labeling. The expressions of cycle related proteins and apoptotic related proteins were determined by Western blot analysis. OA strongly inhibited human hepatoma cells proliferation. When Bel-7402 cells were pretreated with OA for 24 h, OA induced apoptosis and G₂/M phase cell cycle arrest in a concentration-dependent manner. Analysis of the cell cycle regulatory proteins demonstrated that OA decreased the protein levels of cyclin B1, but increased the protein levels of p-Cdk1 (Tyr15) and p-Cdc25C (Ser 216). Moreover, OA modulated the phosphorylation of protein kinases Chk1 and p2l. Western blotting assay also showed significant decrease of Bcl-2 protein expression and increase of Bax protein expression, the cytosol Cyt c level, cleaved-caspase-9 and cleaved-caspase-3 activity. These data suggest that OA produces anti-tumor effect via induction of G₂/M cell cycle arrest and apoptosis.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; G2 Phase Cell Cycle Checkpoints ; drug effects ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; M Phase Cell Cycle Checkpoints ; drug effects ; Oleanolic Acid ; pharmacology

OBJECTIVETo explore the effect of Shenxiong Huayu Capsule (SHC) on the cerebral ischemia-reperfusion (IR) injury and the expression of growth associated protein 43 (GAP43) after total cerebral IR in the hippocampal CA1 region of rats.

METHODSTotally 100 male adult SD rats were randomly divided into five groups, i.e., the control group, the model group, the group A (by taking SHC once daily), the group B (by taking SHC twice daily), and the group C (by taking SHC thrice daily), 20 in each group. The total IR model was prepared by improved Pulsinelli's 4-vessel occlusion method. Morphological changes of the hippocampal CA1 region were observed by HE staining at day 1, 3, 7, and 14. The expression of GAP43 in the hippocampal CA1 region was detected using immunohistochemical assay at day 1, 3, 7, and 14. Meanwhile, the behavioral score was determined. The expression of GAP43 in the hippocampal CA1 region was detected using Western blot at day 14.

RESULTSCompared with the control group, the expression of GAP43 increased in the model group, the behavioral score was elevated, degenerated neurons increased, and survival neurons decreased in the model group (all P < 0.05). Compared with the model group, the expression of GAP-43 increased (with the most significant difference seen in the group C, P < 0.01), the behavioral score significantly decreased, degenerated neurons decreased, and survival neurons increased in each HSC group (all P < 0.05). Survival neurons obviously increased at day 14, of which, most number of survival neurons and highest contents of GAP43 protein could be seen in the group C, showing statistical difference when compared with those of the group A and the group B (P < 0.01).

CONCLUSIONSHC had protective effect on total cerebral IR in the hippocampal CA1, which might be associated with increased expression of GAP43.

Animals ; Brain Ischemia ; metabolism ; CA1 Region, Hippocampal ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; GAP-43 Protein ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

Objective To investigate the effect of salvia mihiorrhiza on spinal cord ischemia reperfusion injury after surgical treatment in patients with cervical canal stenosis. Methods Retrospective analysis of 64 cases had cervical canal stenosis in the last 5 years in our hospital. Sixty-four cases were randomly divided into the salvia mihiorrhiza group(31 cases)and the control group(33 cases). The therapeutic effect was assessed using JOA grade system. Results In the salvia mihiorrhiza group,the JOA average score was 8. 8 ±2. 6 before surgical treatment, after two weeks of surgical treatment it was 13. 7 ± 2. 4. The JOA improvement ratio was (61. 5 ± 2. 9) % . In the control group,the JOA average score was 9. 1 ±2. 2 before surgical treatment,after two weeks of surgical treatment it was 13. 4 ± 2. 3. The JOA improvement ratio was (60. 5 ± 2.2)% .The JOA improvement ratio in the salvia mihiorrhiza group was significantly higher than that in the control group (P < 0. 05) . Conclusions Salvia mihiorrhiza has protective effect on spinal cord ischemia reperfusion injury.

AIM:To investigate the feasibility of using human amniotic epithelial cells(HAECs) sheets,which are cultured by using a biodegradable fibrin sealant to re-establish corneal surface layer.METHODS:Human amniotic membrane(AM) harvested at the time of elective caesarean section was digested by collagenase and trypsin respectively to obtain HAECs.The HAECs were planted on Millicell culture dishes coated with biodegradable fibrin glue,and cultured by using the air-lifting cultivation.The HAECs sheets were investigated morphologically by Hematoxylin-Eosin staining,scanning electron microscopy(SEM) and were identified by cytokeratin immunohistochemistry.RESULTS:HAECs sheets were transparent and successfully cultured by using a biodegradable fibrin sealant.Most of the cultured cells were round or polygon and typical slabstone-like appearance.Many microvilli were observed on cell surfaces by SEM.Cytokeratin staining was positive.HAECs had stratificated growth tendency and became corneal-like stratificating epithelial cells.CONCLUSION:A commercially available fibrin sealant was an effective means of tissue engineering to create a carrier-free,transplantable HAECs sheets.The sheets were potential graft to re-establish corneal surface layer.

Objective To observe the effects of diazepam on the percen-tage of δ -band power of quantitative pharmaco -electroencephalogram ( QPEEG ) in rabbits and the relation with benzodiazepine receptor.Methods Thirty-six healthy rabbits were randomly divided into 6 groups (each n=6),where rabbits in each group were intravenously injected with 1 mL· kg-1 0.9%NaCl (blank group), 0.12 mg· kg-1 , 0.25 mg· kg-1 or 0.50 mg· kg-1 diazepam groups, 0.1 mg· kg-1 flumazenil group and 0.1 mg· kg-1 flumazenil +0.5 mg· kg-1 diazepam ( combined group ).Then a digital EEG system was adopted to record the changes of QPEEG in eight encepha-lic regions before and after injection at each time point.Results The per-centage of δband power in eight encephalic regions of 0.25, 0.50 mg· kg-1 diazepam groups are noted increase within 0.5 to 30 min.The change of the percentage of δ band power has a positive correlation with diazepam dose.Compared with the 0.5 mg· kg-1 diazepam group,the percentage of δ-band power in combined group was lower at same time point.Conclusion Diazepam increases the percentage ofδ-band power of QPEEG in the way of dose dependence in rabbits.It indicates that benzodiazepine receptor mediates the effect of diazepam and the percentage of δ-band power may become an index that reflects the degree of sedation and hypnosis.

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.

ObjectiveTo analyze the prevalence and epidemic trend of human brucellosis between 2007 -2010 in Qian'an county,and to provide a basis for future prevention and control measures. Methods Epidemiological data of brucellosis between 2007 - 2010 were obtained from the national disease surveillance report on management information system, population data were from the national disease surveillance information management system for basic information reporting system, and descriptive epidemiological methods was used for statistical analysis. Results A total of 86 cases were infected with brucellosis in Qian'an county from 2007 to 2010, with 68 cases of male and 18 cases of female, and male to female ratio was 3.78 : 1. Patients were mainly in the age of 20 - 55, with 40 - ＜ 45 year-old group the highest. All patients were local residents. Most professional were livestock acquisition, processing and aquaculture personnel. The majority of patients lived in Jianchang town,with 51 cases, accounting for 59.30％. Thirteen patients were reported in 2007, incidence was 0.0201‰(13/ 647 983). Thirty-nine patients were reported in 2010, and incidence was 0.0563‰(39/657 380). There was a fluctuations increase from 2007 to 2010. In 2007 brucellosis occurred in only two townships(towns), which spread to eight townships (towns) and urban areas in 2010. ConclusionsThe epidemic of human brucellosis in Qian'an is in a spreading trend. We recommend the government to strengthen the quarantine of livestock, and the infected livestock should be timely treated. Strengthen the prevention and control in Jianchangying. At the same time increase the brucellosis propaganda, and enhance self-protection awareness of the occupational groups.

Objective To investigate MR imaging features of femoral marrow in treated ?-thalassemia major.Methods MR imaging of the proximal femoral marrow was performed in 35 cases of ?-thalassemia major and 45 age-and sex-matched normal children as control.Coronal images of femoral marrow with the techniques of spin echo and fast field echo(FFE)were obtained.On T_1-weighted imaging the red and yellow femoral marrow were judged and marrow distribution was classified into five groups.The hemosiderosis of marrow was judged on the basis of signal intensity of marrow on FFE imaging.The marrow distribution classification and the hemosiderosis on MR imaging were correlated with clinical features.Results On FFE,marrow hemosiderosis occurred in 15 patients with a marked hypo-intensity signal and was related to the age(P=0.032).On T_1-weighted imaging,the femoral marrow in 35 patients was classified as groupⅢand IV,while the marrow distribution was groupⅠorⅡin all normal children,there was statistically significant difference(P

Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P＜0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P＜0.01) and 27.9%(χ2=42.68,P＜0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.

OBJECTIVETo observe clinical therapeutic effect of acupuncture in abatement of fever.

METHODSEighty-six cases of high fever were randomly divided into two groups, an acupuncture group (n = 45) treated with acupuncture at Dazhui (GV 14) and a drug group (n = 41) treated with intramuscular injection of Antongding Injectio. Changes of body temperature before and after 24 hours treatment were observed by single blind.

RESULTSIn the acupuncture group, 9 cases were cured, and in the drug group, 2 cases were cured, the acupuncture group was better than the drug group in abatement of fever.

CONCLUSIONAcupuncture has a reliable therapeutic effect on fever.

Acupuncture Points ; Acupuncture Therapy ; Common Cold ; Fever ; Humans ; Single-Blind Method