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

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; pathology ; Female ; Humans ; Laryngeal Neoplasms ; pathology ; Lymphatic Metastasis ; pathology ; Lymphatic Vessels ; pathology ; Male ; Middle Aged ; Neck ; pathology ; Neoplasm Staging

Objective To evaluate the efficacy and safety of methotrexate(MTX)on the hip involve ment in patients with ankylosing spondylitis(AS).Methods Among the AS patients with hip joint involvement admitted to the department from 1999—2002,50 patients were treated with sulfasalazine(SSZ)(control group, CG)and 48 MTX(observation group,OG).One kind of NSAIDs was taken by the patients in both groups as the basic therapy.The treatment was maintained in both groups after being discharged.Patients were followed-up for three years.The observation parameters included symptoms,signs,Bath AS disease activity index(BASDAI), Bath AS functional index(BASFI),hip joint function score,the CT staging of hip joint involvement,serum in- flammation markers,laboratory tests and side effects.The data were analyzed with SPSS10.0 statistics software. Results At the 1st,2nd and 3rd year,44,38 and 32 cases in the OG and 45,38 and 31 cases in the CG were followed up respectively.During the three-year follow-up period,the hip joint function score of the OG was significantly higher than that of the CG(P

OBJECTIVE: To optimize the preparation process of Xiangsha Liujun Granule. METHODS: By the orthogonal design. taking the rates of recovery of volatile oil, Ginsenoside Re and qualified granules as the indexes, we optimized the oil extracting conditions, the decocting conditions and the conditions of granulation by spayer. RESULTS: The optimum oil ex- tracting conditions for four kinds of herbs(Pericarpium Citri Reticulatae etc.) were that water was 30 times of herbs in weight, soaking time was 3 hours and extracting time 15 hours. The optimum decocting conditions for four kinds of herbs(Fuling etc.) were that water was 15 times of herbs in weight, soaking time was 3 hours and decocting time 4 hours, herbs were decocted twice. The optimum conditions of granulation by sprayer were that the fliud extract was 2 times of Dextrin in weight, the rel- ative density of the fliud extract was 1. 30(measured at 25℃), the voltage of sprayer was 120 volts, the period of shaking bag was 65 times(shaking bag back and forth twice in each period). CONCLUSION: The results of the experiment make it clear that the preparation process of Xiangsha Liu jun Granule is stable and feasible.

RT-PCR amplification of ginseng ?-amyrin synthase gene was successfully performed based on the total RNAs extracted from ginseng hairy roots induced by Agrobacterium rhizogenes A4 using a modified guanidine isothiocyanate-method. Sequence analysis of this gene revealed that its sequence was consistent with the sequence of a previously reported ginseng ?-amyrin synthase gene (GenBank No. AB009030). This gene was inserted into pMD-119T simple vector and transformed into E.coli DH5?. Furthermore antisense plant expression vector of this gene was constructed using the pBI121 vector, laying foundation for studies on antisense regulation of ginseng ?-amyrin synthase gene.

Environmental Pollution ; prevention & control ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Iatrogenic Disease ; prevention & control ; Medical Laboratory Personnel ; psychology

Objective To comparatively analyze serum levels of zinc , iron, magnesium, calcium, copper and phosphorus and their relationships with serum albumin and circulating HCV viral load in patients with chronic HCV infection , subjects with spontaneously resolved HCV infection and healthy subjects . Methods Serum levels of the six trace elements in patients with chronic HCV infection (n=59), subjects with spontaneously resolved HCV infection (n=65) and healthy subjects (n=48) were measured by a flame atomic absorption spectrophotometer and then a comparative analysis was performed to analyze the differences among the three groups .The relationships of the six trace elements with serum albumin and HCV viral load were analyzed among patients with chronic HCV infection .Results Compared with healthy subjects , the levels of serum zinc were significantly decreased , but serum levels of iron , copper , phosphorus were signifi-cantly increased in patients with chronic HCV infection .There was a significant positive correlation between the levels of zinc(r=0.4022, P=0.0016)and albumin in patients with chronic HCV infection with ALT no less than 40 IU/L, whereas negative correlations were presented between trace elements of iron ( r=-0.3001, P=0.0209), copper (r=-0.3856, P=0.0036), phosphorus (r=-0.3600, P=0.0075) and serum albumin.The circulating HCV viral load was negatively correlated with serum zinc (r=-0.4367, P=0.0005), but positively correlated with serum copper (r=0.3328, P=0.0139).The serum levels of six trace elements showed no significant differences between healthy subjects and spontaneous resolvers of HCV infection.Moreover, no significant differences of serum calcium and magnesium were found among the three groups.Conclusion Chronic HCV infection can induce abnormal serum levels of zinc , iron, copper and phosphorus and the abnormal serum levels of trace elements were closely related with liver function and HCV viral load.With the spontaneous clearance of HCV infection , the serum levels of trace elements could restore to normal .

Fomentation engineering is one of the modern biotechnologies. It has been intensively studied and widely applied in edible fungi. Based on the review of research history of liquid fermentation for edible fungi, the research status of liquid fermentation about edible fungi were summarized, and its application prospects on edible fungi production of our country were described in this paper.

Genetic engineering is a powerful tool in Panax ginseng breeding.Genetic transformation and plant regeneration are the premise and foundation involved in genetic engineering of Panax ginseng.Ginseng can be regenerated through organogenesis or somatic embryogenesis and indirect somatic embryogenesis is mainly used for its regeneration.Summurized the factors influencing plant regeneration such as different explants,different carbohydrates,somatic embryo optimization and hormone-free approach.Ginseng transformation has been achieved by Agrobacterium tumefaciens and Agrobacterium rhizogenes and transgenic ginseng with good characters was obtained by introducing genes associated with biosynthesis of ginsenosides or herbicide gene.Hairy root culture system can supply large scale of ginsenosides,thus effect of rolC genes on ginseng hairy root induction,regeneration and bioreactor culture of hairy root were discussed.Additionally,problems that are present in genetic engineering of Panax ginseng were also discussed in this review.

Objective To discuss the histology change,apoptosis state and Bcl-2,Bax expression after the bone cement leakage into the intervertebral disc in vertebroplasty.Methods Eight majority canis familiaris were studied.Three lumbar intervertebral discs(L2 to L5)in each dog were randomly classified into three groups(control group,PMMA group,and CPC group),the canine discs were stabbed by 18-gauge needle,and 0.1 ml cement was injected into them.Control discs were only stabbed and injected with nothing.Histology of all discs was studied 24 weeks after the operation.Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and immunohistochemisty were used to detect apoptosis and Bcl-2,Bax expression in the discs.The data were statistically analyzed by SPSS 12.0.Results Intervertebral disc degeneration was not found in control groups.In bone cement groups,however,ruptured or serpentine patterned fibers,decreased cellularity of the nucleus pulposus and condensed matrix of the nucleus pulposus were found in histologic results.The Bax protein decreased in the order of control group, CPC group,and PMMA group.However,the Bcl-2 protein increased in the order of control group,CPC group,and PMMA group.The histology grade was significantly different among the three groups under ANOVA analysis(P