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.

Equipment Design ; Facial Injuries ; therapy ; First Aid ; instrumentation

OBJECTIVETo observe clinical therapeutic effect of mild-warm moxibustion on diabetic peripheral neuropathy (DPN) and to probe the mechanism.

METHODSSixty cases of DPN were randomly divided into a mild-warm moxibustion group, an acupuncture group and a medication group, 20 cases in each group. In the mild-warm moxibustion group and the acupuncture group, the same points, Shenshu (BL 23), Pishu (BL 20), Zusanli (ST 36), Yongquan (KI 1), etc. were selected; and the medication group were treated with Mecobalamin tablets. Their therapeutic effects and changes of fasting blood-glucose (FBG), glycosylated hematoglobin (GHB), hemorheological indexes, plasma endothelin (ET), nitric oxide (NO) and malondialdehyde (MDA) before and after treatment were investigated.

RESULTSThe total effective rate was 90.0%, FBG, GHB, hemorheological indexes, plasma ET, NO and MDA significantly improved in the mild-warm moxibustion group (P < 0.01), with no significant difference as compared with those in the acupuncture group (P > 0.05), but with a significant difference as compared with the medication group (P < 0.05).

CONCLUSIONMild-warm moxibustion has definite therapeutic effect on diabetic peripheral neuropathy, which is better than that of Mecobalamin.

Adult ; Aged ; Diabetic Neuropathies ; physiopathology ; therapy ; Endothelin-1 ; blood ; Female ; Hemodynamics ; Humans ; Male ; Malondialdehyde ; blood ; Middle Aged ; Moxibustion ; methods ; Nitric Oxide ; blood ; Peripheral Nervous System Diseases ; physiopathology ; therapy

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

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.

In lieu of large samples of cases and/or controls with hundreds of markers spreading throughout the human genome, researchers started to notice the dramatic increase of genome-wide association study (GWAS) for complex disorders, in the last 5 years. This paper highlights the statistical challenges in such huge-scale genetic studies, and introduces the analytical strategies and steps for handling GWAS data. Such issues as quality control of data, population stratification, methods available to data analysis and results presentation, replication, as well as the limitations of GWAS studies and the challenges presenting for statistics, are addressed.

China Three Gorges University(CTGU) started undergraduate medical education for foreign students since 2004. Over the years,through continuous interaction with coordinators,CTGU grad-ually transited from passive teaching to active teaching. Firstly we determined basic principles,objectives and requirements of talent training programme. Secondly,we strengthen Chinese language teaching, reasonably adjusted professional curriculum and teaching content,added basic medical courses and rea-sonably arranged clinical practice based on the concrete situation in our country. in order to make the cur-riculum system consistent with the require-ments of talent training.