Objective To investigate the expression and the effects of X-chromosome linked inhibi- tor of apoptosis (XIAP) associated factor 1 (XAF1) on apoptosis and cell proliferation in SMMC7721 hepatocellur carcinoma (HCC) cell line.Methods The expression of XAF1 mRNA and protein in hu- man SMMC7721 cell line were detected by semi-quantitative,RT-PCR and Western blot.Plasmid con- structs expressing sense and antisense XAF1 were generated and transfected into SMMC7721 cell line to establish stable transfectants.Cell proliferation and apoptosis were detected by MTT,flow cytometry and TUNEL.Results XAF1 mRNA and protein were detectable in SMMC7721 cell line but lower than that in normal liver cell.Stable expression of XAF1 significantly inhibited cell proliferation and increased spontaneous apoptosis in SMMC7721 cell (P＜0.05).Over-expression of XAF1 in stable transfactants increased the sensitivity of SMMC7721 cell to chemotherapeutic drugs such as 5-fluorouracial and hydroxycamptothecin.Conclusions Over-expression of XAF1 induces apoptosis and inhibits SMMC7721 cell growth.XAF1 may be a promising candidate for HCC gene therapy.

OBJECTIVETo study the conditions on separation and regeneration of protoplast from Phellinus igniarius.

METHODThe effects of enzymolysis conditions of P. igniarius mycelia on yield of protoplast and culturing conditons on regeneration ratio of protoplast were investigated.

RESULTWhen the 8 days-old mycelia was hydrolysed by 1.5% of lywallzyme adding to driselase of 0. 5% and at 30 degrees C for 3 h and enzymolysis was stablized by sucrose as a stablisher of osmotic pressure, higher yield of P. igniarius protoplast was obtained. If 10 days-old mycelia was used as raw material of enzymolysis and manntol was selected as stablisher of osmotic pressure of enzymolysis, higher regeneration ratio of P. igniarius protoplast also would be obtained in following regeneration step at same time keeping higher yield. For the regeneration processing, it was beneficial for the regeneration of P. igniarius protoplast that PDA plusing mulberry ramulus was used as the culture medium of regeneration and manntol was selected as the osmotic pressure establisher of regeneration culture medium.

CONCLUSIONThe method and conditions to keep both higher yield and regeneration ratio of P. igniarius protoplast were obtained.

Culture Media ; pharmacology ; Fungal Proteins ; pharmacology ; Glucan Endo-1,3-beta-D-Glucosidase ; pharmacology ; Glycoside Hydrolases ; pharmacology ; Mannitol ; pharmacology ; Multienzyme Complexes ; pharmacology ; Osmotic Pressure ; Peptide Hydrolases ; pharmacology ; Polyporaceae ; drug effects ; physiology ; Protoplasts ; drug effects ; physiology ; Regeneration ; drug effects ; Sucrose ; pharmacology ; Temperature

Objective: To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotocin-induced diabetic rats. Methods: The chemical constituents in Callicarpa nudiflora extract were identified by UPLC-Q-TOF-MS. Callicarpa nudiflora extract (0.15 and 0.3 g/kg) was orally administered to streptozotocin-induced diabetic rats for 42 d. The effects of Callicarpa nudiflora extract on body weight, blood glucose, serum insulin, total cholesterol, triglyceride, LDL-C and HDL-C were investigated, and its effect on liver and pancreatic pathology was assessed by histopathological analysis. Moreover, the expression levels of adenosine 5'-monophosphate-activated protein kinase (AMPK), glucose transporter type 4 (GLUT4), phospho-AMPK/AMPK, and p-acetyl-coA carboxylase (P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry. Results: A total of 34 compounds, including 8 iridoids, 14 phenylpropanoids, and 12 flavonoids, were identified from Callicarpa nudiflora extract. Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels, including serum insulin, total cholesterol, triglyceride, LDL-C, and HDL-C. Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes. Furthermore, Callicarpa nudiflora extract increased the expression levels of phospho-AMPK, GLUT4, P-ACC, and insulin receptor substrate-1 and decreased the expression level of PPAR毭 in diabetic rats.Conclusions: Callicarpa nudiflora extract improved oral glucose tolerance, lipid metabolism, insulin resistance, and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.

Warfarin is prone to cause thromboembolism or bleeding due to its narrow therapeutic window.Studies have shown that both genetic and environmental factors affect individual difference of warfarin dose.This review will summarize the recent progress of models building and valida-ting at home and abroad to provide a reference for individualized treat-ment of warfarin.

Antithrombotic drugs play an important role in the prevention and treatment of thromboembolism , which include anticoagulant , anti-platelet therapies and thrombolytic drugs.In this paper , we review the pharmacological properties of these most commonly used oral antithrom-botic drugs and explore the development of anticoagulant and antiplatelet therapies , in order to guide the safety and rational use of antithrombotic drugs in clinic.

OBJECTIVETo explore the expression and clinical significance of P-glycoprotein (P-gp)/mdr1mRNA, multidrug resistance-associated protein (MRP) and lung resistance protein (LRP) in newly diagnosed non-Hodgkin's lymphoma.

METHODSmdr1 mRNA of in 41 patients with non-Hodgkin's lymphoma was assayed by semi-quantitative RT-PCR. The expressions of P-gp, MRP and LRP proteins in lymph node viable blasts were identified by flow cytometry. The results were compared with those obtained from control cases, and the correlation of the changes with clinical outcomes was analyzed.

RESULTS(1) Among the 41 cases, the positive expression of P-gp protein was detected in 8 cases, MRP in 7 cases, LRP in 15 cases, and mdr 1 mRNA in 11 cases. (2) The P-gp and LRP levels in NHL were significantly higher than those in control group, but MRP wasn't. The P-gp over-expression was significantly associated with mdr1mRNA (r = 0.396, P = 0.01). No correlation was showed among the expressions of P-gp, MRP and LRP. (3) Patients with P-gp expression had a poorer outcome of chemotherapy than those with P-gp-negative (P = 0.005). P-gp expression was significantly associated with higher clinical stage (P = 0.046) and elevated serum lactate dehydrogenase level (P = 0.032), but not associated with malignant degree (P = 0.298). MRP had no impact on the outcome of chemotherapy (P = 0.212), and wasn't significantly associated with higher clinical stage (P = 0.369), elevated LDH (P = 0.762) and higher malignant degree (P = 0.451). Patients with LRP expression had a poorer outcome of chemotherapy than those LRP-negative (P = 0.012). LRP expression was significantly associated with higher clinical stage (P = 0.0019), elevated LDH (P = 0.02) and higher malignant degree (P = 0.01).

CONCLUSIONThe data of this study indicate that P-gp and LRP expressions but not MRP expression are important in the mechanism of drug resistance associated with a poor clinical outcome in previously untreated NHL.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cell Line, Tumor ; Child ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Humans ; Lactate Dehydrogenases ; blood ; Lymph Nodes ; metabolism ; Lymphoma, Non-Hodgkin ; drug therapy ; metabolism ; pathology ; Male ; Middle Aged ; Multidrug Resistance-Associated Proteins ; metabolism ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Remission Induction ; Vault Ribonucleoprotein Particles ; metabolism ; Young Adult

OBJECTIVETo investigate the association of advanced oxidation protein products (AOPP) with oxidative stress in colon cancer cells exposed to intermittent hypoxia (IH).

METHODSColon cancer SW480 cells were exposed to IH, continuous hypoxia, or normoxia. Enzyme-linked immunosorbent assay (ELISA) was employed to examine the levels of AOPP and vascular endothelial growth factor (VEGF), xanthine oxidase assay was used to determine malonaldehyde (MDA) and glutathione peroxidase (GSH-PX), and Western blotting and immunofluorescence assay were performed for detection of transforming growth factor-β(1) (TGF-β(1)) expression.

RESULTSCompared with the normoxia group, the two hypoxia groups showed significantly increased AOPP and MDA levels (P<0.05) and lowered SOD and GSH-PX levels (P<0.05). The concentration of AOPP was positively correlated to MDA, VEGF, and TGF-β(1) levels (P<0.05), but inversely to SOD. No significant correlation was found between AOPP and GSH-PX levels.

CONCLUSIONCompared with continuous hypoxia, IH results in more obvious protein oxidation in relation to oxidative stress. The increased expression of VEGF and TGF-β(1) in the context of hypoxia is closely related to AOPP level.

Advanced Oxidation Protein Products ; metabolism ; Cell Hypoxia ; Cell Line, Tumor ; Colonic Neoplasms ; metabolism ; Humans ; Oxidative Stress ; Transforming Growth Factor beta1 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Objective Through observing the morphological changes of the prepared influenza viruses (H1N1) treated with the different concentration of Nonidet P-40 solutions and added with antibody against the influenza virus using transmission electron microscopy (TEM), to explore the principle of application of the internal antibody for immunoassay of influenza virus. Methods Through treating the virus samples with serial diluted Nonidet P-40 solutions from 0.01% to 0. 2% and/or not adding antibody against the influenza virus, then investigating the samples by TEM to obtain the morphological changes of the virions. Results The serial images show that the denudation degree of the virions is proportional with the rise of NP-40 concentration, and partly denuded virion image appeared at 0. 1% NP-40 treatment, also under this condition no influence was observed on antibody binding to the virus. Conclusion This work demonstrated the interaction between influenza virion and its antibody under nonionic surfactants existing,which supports the advantages of sample preparation for immunoassay enveloped virus using internal antibody theoretically.

OBJECTIVETo detect methylation in promoter region of hMSH2 gene in esophageal cancer.

METHODSSpecimens of cancer and normal tissues freshly removed from 32 cases of esophageal cancer patients without previous radiotherapy, chemotherapy or other treatment were preserved at -80 degrees C within 30 min. Methylation specific PCR (MSP) was used to detect methylation of mismatch repair gene (MMR) hMSH2 in promoter region in esophageal cancer and normal esophageal tissues.

RESULTSThe frequencies of methylation of hMSH2 gene in promoter region of cancer and normal esophageal tissues were 32.4% (11/32) and 0/30 (0%), respectively, and significant difference was found between the two groups (P < 0.01). The frequency of methylation in elder patients (> or = 70 years old) was significantly higher than that in younger patients (< 70 years old) (P < 0.05). Methylation was less frequently found in grade I-II (18.2%) than in grade III-IV (70.0%) (P < 0.05).

CONCLUSIONMethylation of hMSH2 gene in promoter region is related to patients' age and histopathological grade of the esophageal cancer.

Aged ; Base Pair Mismatch ; Carcinoma, Squamous Cell ; genetics ; pathology ; DNA Methylation ; Esophageal Neoplasms ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; MutS Homolog 2 Protein ; genetics ; Promoter Regions, Genetic ; Transfection

To investigate the effects of c-myc antisense oligodeoxynucleotide (ASODN) on the telomerase activity and the induction of apoptosis in HL-60 cells, and to explore the relationship between the telomerase activity and the expression of c-myc gene in HL-60 cells, after treatment by c-myc ASODN, the expression of c-myc was detected by RT-PCR, the apoptosis, cell cycle were detected with agarose gel electrophoresis and flow cytomety, and the telomerase activity was determined with TRAP-ELISA. The results showed that after blocking c-myc gene with ASODN for 72 hours, it is obvious that the expression of c-myc gene was inhibited. The percentage of S phase HL-60 cells decreased from 55.6% to 30%, the early apoptosis peak appeared (the percentage of apoptosis cells were 25.2%) and the DNA ladders were shown. OD(450 - 690) were 2.648 +/- 0.42, 2.324 +/- 0.36, 2.162 +/- 0.38, 1.952 +/- 0.14, 1.805 +/- 0.40, 1.616 +/- 0.41 and 2.466 +/- 0.29, respectively, as the cells were treated with 0, 1, 2, 3, 4, 5 micromol/L ASODN and 5 micromol/L SODN for 72 hours. The difference was significant when compared 3, 4, 5 micromol/L groups with 0 micromol/L ASODN group respectively (P < 0.05), but the difference was no significant when compared 1, 2 micromol/L ASODN and 5 micromol/L SODN groups with 0 micromol/L ASODN group (P > 0.05). It is concluded that the c-myc gene ASODN may induce the apoptosis of cells, inhibit cells from G(1) phase into S phase and regulate the telomerase activity down in HL-60 cells by blocking the expression of c-myc gene.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; HL-60 Cells ; Humans ; Oligonucleotides, Antisense ; genetics ; pharmacology ; Proto-Oncogene Proteins c-myc ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; metabolism