OBJECTIVETo observe the effect of bear bile powder (BBP) on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma (HCC) xenograft, and to explore its mechanism for treating HCC.

METHODSThe subcutaneous xenograft model was established using HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly divided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were administered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT-mediated dUTP nick end labeling (TUNEL). The expression of Bcl2-associated X protein (Bax), B cell lymphoma/eukemina-2 (Bcl-2), cyclin-dependent protein kinase (CDK4), cyclinD1 were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of signal transducers and transcription activators 3 (p-STAT3), proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry.

RESULTSBBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group (P < 0.01). Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma carcinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-regulate mRNA expression of Bcl-2, CDK4, and cyclinD1. Immunohistochemical results showed that BBP could up-regulate the expression of Bax and inhibit the protein expression of p-STAT3, PCNA, Bcl-2, CDK4, and cyclinD1.

CONCLUSIONBBP could induce the apoptosis of hepatoma carcinoma cells and inhibit their proliferation by regulating STAT3 pathway.

Animals ; Bile ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Ursidae ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo evaluate the effect of bear bile powder (BBP) on angiogenesis, and investigate the underlying molecular mechanisms.

METHODSA chick embryo chorioallantoic membrane (CAM) assay was used to evaluate the angiogensis in vivo. Human umbilical vein endothelial cells (HUVECs) were treated with 0, 0.25, 0.5, 0.75, and 1.0 mg/mL of BBP for 24, 48 and 72 h, respectively. The 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the viability of HUVECs. Cell cycle progression of HUVECs was examined by fluorescence-activated cell sorting (FACS) analysis with propidium iodide staining. HUVEC migration was determined by wound healing method. An ECMatrix gel system was used to evaluate the tube formation of HUVECs. The mRNA and protein expression of vascular endothelial growth factor (VEGF)-A in both HUVECs and HepG2 human cells were examined by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively.

RESULTSCompared with the untreated group, BBP inhibited angiogenesis in vivo in the CAM model (P< 0.01). In addition, treatment with 0.25-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability by 14%-27%, 29%-69% and 33%-91%, compared with the untreated control cells (P< 0.01). Additionally, BBP inhibited the proliferation of HUVECs via blocking the cell cycle G to S progression, compared with the S phase of untreated cells 48.05%± 5.00%, 0.25-0.75 mg/mL BBP reduced S phase to 40.38%± 5.30%, 36.54± 4.50% and 32.13± 3.50%, respectively (Pglt; 0.05). Moreover, BBP inhibited the migration and tube formation of HUVECs, compared with the tube length of untreated cells 100%± 12%, 0.25-0.75 mg/mL BBP reduced the tube length to 62%± 9%, 43%± 5% and 17%± 3%, respectively (p< 0.01). Furthermore, BBP treatment down-regulated the mRNA and protein expression levels of VEGF-A in both HepG2 cells and HUVECs.

CONCLUSIONBBP could inhibit the angiogenesis by reducing VEGF-A expression, which may, in part, explain its anti-tumor activity.

Animals ; Bile ; chemistry ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Gene Expression Regulation ; Hep G2 Cells ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Neovascularization, Physiologic ; Powders ; RNA, Messenger ; genetics ; metabolism ; Ursidae ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate the angiogenic effect of the Xiongshao capsule (XSC) in human umbilical vein endothelial cells (HUVEC), and to investigate the possible molecular mechanisms mediating its biological effect.

METHODSSerum pharmacology was applied in this study, in which different doses of XSC were administrated to rats orally and then XSC-containing serum (XSC-S) was collected for the following in vitro experiments. The viability of HUVEC was determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell density was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with propidium iodide staining was performed to determine cell cycle phase. Cell migration was determined by wound-healing method. Capillary tube formation by HUVEC was examined using ECMatrix gel-based assay. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression levels were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbant assay (ELISA) analyses.

RESULTSXSC-S dose-dependently stimulated proliferation of HUVEC by promoting the cell cycle G1 to S progression. In addition, XSC-S treatment dramatically increased the migration and capillary tube formation of HUVEC in a dose-dependent manner. Moreover, XSC-S enhanced the expression of VEGF and bFGF at both mRNA and protein levels.

CONCLUSIONXSC can promote several features of angiogenesis in endothelial cells through up-regulating the expression of bFGF and VEGF, suggesting that XSC may be a potential novel therapeutic agent for the treatment of ischemic heart diseases.

Animals ; Capsules ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Collagen ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Laminin ; pharmacology ; Male ; Neovascularization, Physiologic ; drug effects ; genetics ; Proteoglycans ; pharmacology ; Rats ; Rats, Sprague-Dawley ; S Phase ; drug effects ; Up-Regulation ; drug effects ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the anti-angiogenic effects of Pien Tze Huang in vivo and in vitro.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 0 mg/mL, 0.25 mg/mL, 0.5 mg/mL, and 1 mg/mL of PZH for 24 h, 48 h and 72 h, respectively. Chicken embryo chorioallantoic membrane (CAM) model was used to evaluate in vivo angiogenesis. An ECMatrix gel system was used to evaluate in vitro angiogenesis by examining the tube formation of HUVECs. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine HUVEC viability. Cell density of HUVECs was observed by phase-contrast microscopy. HUVEC migration was determined by wound healing method. The mRNA and protein expression of vascular endothelial growth factor A (VEGF-A) and basic fibroblast growth factor (bFGF) in both HUVEC and human colon adenocarcinoma cells (HT-29) was examined by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immune sorbent assay (ELISA), respectively.

RESULTSPZH treatment significantly reduced the total number of blood vessels compared with the untreated control in the chicken embryos and resulted in a significant decrease in capillary tube formation and cell density of HUVECs (P<0.05). In addition, treatment with 0.25-1 mg/mL of PZH for 24 h, 48 h, and 72 h respectively reduced cell viability by 9%-52%, 24%-87% or 25%-87%, compared with the untreated control cells (P<0.05). Moreover, PZH treatment decreased the migration of HUVECs. Furthermore, PZH dose-dependently suppressed the expression of VEGF-A and bFGF on both mRNA and protein levels (P<0.05).

CONCLUSIONPZH could inhibit angiogenesis in vivo in CAM model and in vitro on HUVECs, suggesting that inhibiting tumor angiogenesis might be one of the mechanisms by which PZH treats cancer.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; HT29 Cells ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Neovascularization, Physiologic ; drug effects ; genetics ; RNA, Messenger ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo determine the influences of Mannose binding protein (MBP) gene polymorphisms on HBV DNA loads and on the progression of liver disease in patients with chronic HBV infection.

METHODThe Codons on 54 MBP gene polymorphisms and HBV DNA loads in a cohort of 395 patients with chronic HBV infection, including 244 with chronic hepatitis B (CHB), 151 with liver cirrhosis (LC) and 88 normal controls were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and fluorescent quantitative PCR (FQ-PCR).

RESULTThe MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC in CHB group showed no significant differences comparing to the normal control group (P > 0.05). The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC on CHB group (severe), compensation phase of LC group and decompensation phase of LC group were higher than those in the normal control group (P < 0.05), the genetic polymorphism of decompensation of LC was 36.5%, highest of all. The MBP genotype frequencies of GGC/GAC and alleles genetic frequencies of GAC of patients with chronic HBV infection were not changed with the differences of HBV-DNA loads.

CONCLUSIONThe codes on 54 MBP gene polymorphisms is not closely related to HBV DNA loads, but was associated with the progression of hepatitis B infection.

Adolescent ; Adult ; Aged ; Cohort Studies ; DNA, Viral ; analysis ; Disease Progression ; Female ; Gene Frequency ; Genotype ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Mannose-Binding Lectin ; genetics ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo evaluate the efficacy and safety of capecitabine as first-line therapy in patients with advanced and recurrent colorectal cancer.

METHODSFrom December 2000 to November 2001, sixty patients with advanced and recurrent colorectal cancer received first-line capecitabine treatment given at a dose of 1250 mg/m(2) twice daily, on days 1 - 14 every 21 days. At least 2 cycles were administered.

RESULTSThe overall response rate was 23.3% with 14 PR, 24 SD (40.0%) and 15 PD. The median survival time was 14.7 months. The survival rate was 63.9% at 12-months and 33.4% at 24-months. Grade III-IV adverse effects were diarrhea in 4 patients (6.6%), anemia in 2 (3.3%) and hand-foot syndrome (HFS) in 1 (1.7%); Grade I-II adverse effects were hyperpigmentation in 20 (33.3%), HFS in 18 (30.0%) and diarrhea in 10 (16.7%).

CONCLUSIONCapecitabine is an efficacious and better-tolerated alternative treatment for the patients with advanced and recurrent colorectal cancer.

Adult ; Aged ; Antimetabolites, Antineoplastic ; therapeutic use ; Capecitabine ; Colorectal Neoplasms ; drug therapy ; mortality ; Deoxycytidine ; adverse effects ; analogs & derivatives ; therapeutic use ; Female ; Fluorouracil ; analogs & derivatives ; Humans ; Male ; Middle Aged ; Survival Rate