Afatinib ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoembryonic Antigen ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gallbladder Neoplasms ; drug therapy ; pathology ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Neoplasm Staging ; Tomography, X-Ray Computed

To observe the effect of various doses of oil of Piper longum unsaponifiable matter (OPUM) to cholesterol gallstones in experimental mice. C57BL/6 mice (n = 60) were randomly divided into 6 groups: control group, model group, OPUM (15, 30 and 60 mg x kg(-1)) group and ursodeoxycholic acid (UDCA, 60 mg x kg(-1)) group, administered for 10 weeks. The level of serum lipid and liver function enzymes were tested. The gallbladder was removed and bile was obtained by centrifugation. Next, the levels of the bile total cholesterol (TC), phospholipid (PL) and bile acid (TBA) were measured. The indicators of lipid peroxidation were determined and cholesterol saturation index (CSI) was calculated. The liver histological changes were observed by HE staining. The results showed that serum TC, TG (triglycerides) and AST (aspartate transaminase) contents, gallbladder cholesterol crystallization and CSI increased significantly (P < 0.05). In addition, the activity of SOD decreased significantly and MDA content increased significantly in liver (P < 0.05). HE staining results showed that the hepatic cord disorder and intracellular lipid droplets increased significantly. All results indicate that lithogenic diet lead to the formation of cholesterol gallstones. In OPUM (30 and 60 mg x kg(-1)) group, serum TC, TG and AST content, gallbladder cholesterol crystallization and CSI decreased significantly, the activity of SOD increased significantly and MDA content decreased significantly. HE staining results showed that OPUM can improve the morphology of liver cell, reduce the degree of hepatic cord disorders and restore the cell morphology close to normal. The cause of OPUM prevents cholesterol gallstone formation maybe due to protect the integrity of the liver cells, lower CSI, and reduce cholesterol crystal formation and hence prevent cholesterol gallstone formation.
Animals ; Aspartate Aminotransferases ; blood ; Bile ; chemistry ; Cholesterol ; blood ; Gallbladder ; Gallstones ; metabolism ; prevention & control ; Liver ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred C57BL ; Piper ; chemistry ; Plant Oils ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Superoxide Dismutase ; metabolism ; Triglycerides ; blood

OBJECTIVETo study the role of arginase-1 (Arg-1) expression in differential diagnosis of hepatocellular carcinoma (HCC), Arg-1 staining pattern in clear cell neoplasm (HCC and non-HCC) and Arg-1 expression in non-hepatocellular tumors.

METHODSSeventy-eight cases of HCC (including 8 cases of clear cell type and 70 cases of non- clear cell type) and 246 cases of non-hepatocellular neoplasms (including 29 cases of metastatic tumors such as breast cancer, nasopharyngeal carcinoma and neuroendocrine carcinoma, 77 cases of tumors with clear cell changes such as malignant melanoma, clear cell renal cell carcinoma and alveolar soft part sarcoma, and 140 cases of other types of tumors such as ovarian endometrioid adenocarcinoma, pituitary tumor and thyroid papillary carcinoma) were studied.Immunohistochemical study for Arg-1 was performed on the paraffin-embedded tumor tissue.

RESULTSIn HCC, Arg-1 demonstrated both cytoplasmic and nuclear staining, with an overall sensitivity of 96.2% (75/78).In well, moderately and poorly differentiated HCC, the sensitivity was 15/15, 100% (41/41) and 86.4% (19/22), respectively. That was in contrast to negative staining for Arg-1 in all the 29 cases of metastatic tumors studied. The sensitivity, specificity, positive predictive value and negative predictive value of Arg-1 in distinguishing HCC from metastatic tumors was 96.2%, 100%, 100% and 90.6%, respectively. Cytoplasmic and membranous staining was observed in clear cell type of HCC. The overall sensitivity of Arg-1 expression in the 77 cases of tumors with clear cell changes was 14.3% (11/77), including 8/15 for malignant melanoma, 2/4 for ovarian clear cell carcinoma and 1/1 gall bladder adenocarcinoma with clear cell component.In malignant melanoma and ovarian clear cell carcinoma, only cytoplasmic staining was demonstrated. There was no expression of Arg-1 in the 140 cases of other tumor types studied.

CONCLUSIONSArg-1 is a sensitive and specific marker for HCC.It is a potentially useful immunohistochemical marker in distinguishing HCC from metastatic tumors. Though also expressed in malignant melanoma and ovarian clear cell carcinoma, Arg-1 shows a different staining pattern as compared with that in HCC.

Adenocarcinoma ; enzymology ; Adult ; Aged ; Arginase ; metabolism ; Carcinoma, Hepatocellular ; enzymology ; pathology ; secondary ; Cell Differentiation ; Diagnosis, Differential ; Female ; Gallbladder Neoplasms ; enzymology ; Humans ; Liver Neoplasms ; enzymology ; pathology ; secondary ; Male ; Melanoma ; enzymology ; Middle Aged ; Ovarian Neoplasms ; enzymology ; Stomach Neoplasms ; enzymology ; pathology

Animals ; Cholestasis ; complications ; Fibrosis ; etiology ; Gallbladder ; pathology ; Humans ; Kidney ; pathology ; Lipid Metabolism ; Liver Cirrhosis ; etiology ; therapy ; Myocardium ; pathology ; Receptors, Cytoplasmic and Nuclear ; physiology

OBJECTIVETo examine the expression of tissue factor pathway inhibitor-2 (TFPI-2) in gallbladder cancer (GBC) and to investigate the anti-cancer activities of TFPI-2 against the growth of GBC.

METHODSTFPI-2 expression in gallbladder normal tissues, gallbladder polyp (GBP) tissues and GBC tissues were examined by reverse transcriptase polymerase chain reaction (RT-PCR), Western blot and immunohistochemical staining. Adenovirus carrying human TFPI-2 gene (Ad5-TFPI-2) were constructed and its anti-cancer effects were investigated in xenograft tumors. Xenograft tumors were constructed by injection of GBC-SD and SGC-996 cells into the flank of nude mice and the volume of xenograft tumors was measured every 3 days until the sacrifice of mice. The apoptosis index of xenograft tumors was examined by TUNEL assay. The status of Bax, Bcl-2 and caspase-3 was examined by Western blot assay.

RESULTSTFPI-2 expression was profoundly lower in GBC tissues (87.0%) when compared to normal tissues (23.3%) and GBP tissues (52.2%; χ(2) = 21.104, P = 0.000). Ad-TFPI-2 significantly inhibited the growth of xenograft tumors in nude mice. Ad-TFPI-2 inhibited GBC-SD cell growth through the induction of apoptosis. The means of total apoptotic cells per field were much higher in Ad5-TFPI-2 group than those in PBS and Ad5-GFP groups. Ad5-TFPI-2 elevated the expression of Bax and cleaved caspase-3, while it decreased the expression of Bcl-2.

CONCLUSIONSTFPI-2 gene and protein was down-regulated in GBC and the down-regulation of TFPI-2 may play a role in the tumorigenesis of GBC. Adenovirus-mediated TFPI-2 can inhibit GBC growth through the induction of apoptosis.

Adenoviridae ; genetics ; Aged ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; therapy ; Genetic Therapy ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Adenocarcinoma ; metabolism ; pathology ; Carcinoid Tumor ; metabolism ; pathology ; surgery ; Child ; Chromogranin A ; metabolism ; Common Bile Duct ; pathology ; surgery ; Common Bile Duct Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Duodenum ; pathology ; surgery ; Gallbladder ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Lymphoma ; metabolism ; pathology ; Male ; Mucin-1 ; metabolism ; Neoplasm Invasiveness ; Rhabdomyosarcoma ; metabolism ; pathology ; Stomach ; pathology ; surgery ; Synaptophysin ; metabolism

BACKGROUNDThe precise molecular mechanisms underlying the gallbladder carcinoma (GBC) metastasis has not been fully elucidated.

METHODSIn the present study, metastasis-associated proteins were identified by comparative proteomic analysis. The functional study of the candidate protein vimentin was further investigated. First, a pair of higher and lower metastatic sublines (termed GBC-SD/M3 and GBC-SD, respectively), originated from the same parental cell line, was screened by spontaneous tumorigenicity and metastasis in vivo in animal study and further characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach comprised two-dimensional gel electrophoresis analysis and mass spectroscopy was used to identify and compare the protein expression patterns between higher metastatic GBC-SD/M3 and lower metastatic GBC-SD cell lines. Then twenty-six proteins were identified.

RESULTSAmong the 26 proteins identified, fourteen proteins were up-regulated and 12 proteins were down-regulated in GBC-SD/M3. Vimentin was identified and found to be overexpressed in GBC-SD/M3 as compared with GBC-SD. This result was further confirmed by quantitative PCR and Western blotting analysis. Furthermore, the cell migration and invasion potency of GBC-SD/M3 in vitro was remarkably suppressed after small interference RNA-mediated knockdown of vimentin. Moreover, immunoblot and immunohistochemical analysis on 12 human GBC specimens showed consistently increased vimentin expression in metastases compared with primary tumors.

CONCLUSIONTumor vimentin level may reflect the pathological progression in some GBC and may be a useful marker for predicting tumor metastasis and a therapeutic target for the treatment of GBC patients with metastases.

Animals ; Blotting, Western ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Electrophoresis, Gel, Two-Dimensional ; Gallbladder Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Neoplasm Metastasis ; genetics ; pathology ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Vimentin ; genetics ; metabolism

OBJECTIVETo study the expression of ephrin-A7 (EphA7) and metadherin (MTDH) and their clinicopathological significances in the benign and malignant lesions of gallbladder.

METHODSEnVisiom immunohistochemical methods was used for determining the expressions of EphA7 and MTDH in routinely paraffin-embedded sections of surgically-resected specimens from 108 cases with gallbladder adenocarcinoma, 15 cases with adenomatous polyp and 35 cases with chronic cholecystitis treated from June 1996 to June 2006. And 46 cases of peritumoral tissues were also harvested as controls (n = 35).

RESULTSThe positive expression rates of EphA7 and MTDH were significantly higher in gallbladder adenocarcinoma than those in peritumoral tissues (χ(2)(EphA7) = 12.65, χ(2)(MTDH) = 13.00; P < 0.01), adenomatous polyp (χ(2)(EphA7) = 8.21, χ(2)(MTDH) = 9.39; P < 0.01) and chronic cholecystitis (χ(2)(EphA7) = 21.21, χ(2)(MTDH) = 23.68; P < 0.01); Moderately-or severely-atypical hyperplasia of gallbladder epithelium was found in the benign lesions with positive expression of EphA7 and/or MTDH. The positive rates of EphA7 and MTDH were significantly lower in the cases of well-differentiated adenocarcinoma, maximal diameter of tumor < 2 cm, no-metastasis of lymph node, and tumor with no-invasiveness of regional tissues than those in the poorly-differentiated adenocarcinoma (χ(2)(EphA7) = 12.34, χ(2)(MTDH) = 12.80; P < 0.01), maximal diameter of tumor ≥ 2 cm (χ(2)(EphA7) = 5.22, χ(2)(MTDH) = 5.00; P < 0.05), cases with metastasis of lymph node (χ(2)(EphA7) = 5.15, χ(2)(MTDH) = 5.86; P < 0.05) and cases with invasiveness of regional tissues (χ(2)(EphA7) = 7.06, P < 0.01; χ(2)(MTDH) = 4.13; P < 0.05) in gallbladder adenocarcinoma (P < 0.05). The high consistency was found between the expressive levels of EphA7 and MTDH in gallbladder adenocarcinoma (χ(2) = 13.11, P < 0.01). The univariate Kaplan-Meier analysis showed that the increased expression of EphA7 (P = 0.023) and MTDH (P = 0.034) was negatively associated with the overall survival. The multivariate Cox regression analysis showed that increased expression of EphA7 and/or MTDH (P(EphA2) = 0.023, P(MTDH) = 0.034) was an independent poor-prognostic predictor for gallbladder adenocarcinoma.

CONCLUSIONSThe expression of EphA7 and/or MTDH might be closely related to the carcinogenesis, progression, clinical biological behaviors and prognosis of gallbladder adenocarcinoma. The positive expression of EphA7 and/or MTDH may predict bad-prognosis in gallbladder adenocarcinoma.

Adult ; Aged ; Cell Adhesion Molecules ; metabolism ; Cholecystitis ; metabolism ; pathology ; Female ; Follow-Up Studies ; Gallbladder ; metabolism ; pathology ; Gallbladder Diseases ; metabolism ; pathology ; Gallbladder Neoplasms ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Polyps ; metabolism ; pathology ; Prognosis ; Receptor, EphA7 ; metabolism

OBJECTIVETo investigate the expression of VEGF-C and VEGF-D and their correlations with lymphangiogenesis and angiogenesis in gallbladder carcinoma.

METHODSFifty cases of gallbladder carcinoma with complete clinical and pathological data were analyzed. The expression of VEGF-C and -D, D2-40, CD31 was assayed by immunohistochemical staining, with 10 samples of normal gallbladder tissues away from cancer and 19 samples of chronic cholecystitis as controls, and their correlation with clinicopathological findings were analyzed retrospectively.

RESULTSThirty-two (64.0%) of the 50 gallbladder cancers were positive for VEGF-C protein expression by immunohistochemistry and the positive rate of VEGF-D protein expression was 62.0% (31/50). The protein expression of VEGF-C and VEGF-D in tumor tissues was significantly higher than that in normal gallbladder tissues away from the tumor (P < 0.05), but no correlation with that in chronic cholecystitis (P < 0.05). The VEGF-C expression correlated with the patient age and lymph node metastasis (both P < 0.05). The VEGF-D expression only correlated with lymph node metastasis (P < 0.05). In the 50 gallbladder cancers, the MLVD was 6.9 + or - 3.6 and the MVD was 36.1 + or - 12.8. The MLVD in both VEGF-C and -D positive groups was significantly higher than that in the negative groups (P = 0.000), and the lymph node metastasis also increased. MVD in both VEGF-C and -D positive groups was higher than that in the negative groups (P < 0.05), and it was also correlated with tumor differentiation (P < 0.05). A significant positive correlation was also found between VEGF-C and VEGF-D expression (r = 0.498, P < 0.01).

CONCLUSIONVEGF-C and VEGF-D are involved in the lymphangiogenesis and angiogenesis in gallbladder carcinoma, promote lymph node metastasis of the tumor, and both are important in the regulation of lymphangiogenesis and angiogenesis in this cancer. VEGF-C and VEGF-D are of clinical significance in evaluating lymph node metastatic potency and estimation of prognosis in gallbladder carcinoma.

Adenocarcinoma ; blood supply ; metabolism ; pathology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Carcinoma, Papillary ; blood supply ; metabolism ; pathology ; Cholecystitis ; metabolism ; pathology ; Female ; Gallbladder Neoplasms ; blood supply ; metabolism ; pathology ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; Retrospective Studies ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo explore the potential molecular targets for diagnosis and treatment of gallbladder cancer by analyzing and comparing the proteomes expressed in human gallbladder cancer and benign gallbladder tissues.

METHODSThe proteins expressed were analyzed using two-dimensional gel electrophoresis. The differentially expressed proteins in tumors were also analyzed by mass spectrometry (MS). AnnexinA3 expression was examined by streptavidin peroxidase immunohistochemical technique on paraffin-embedded tissue sections from 50 patients of gallbladder cancer and 38 cases of chronic eholecystitis.

RESULTSProtein extracts of individual sample in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the tissue samples of gallbladder cancer. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors and the other eight proteins were underexpressed. The positive expression rates of annexinA3 in gallbladder cancer was significantly higher than that in chronic cholecystitis, and the difference was statistically significant (74.0% vs 21.1%, P < 0.01). In the gallbladder cancer, no correlation was obtained between annexinA3 and age, gender or histologicl type (P > 0.05), but overexpression of annexinA3 correlated significantly with those cases with a lower histological grading (40.0% vs 82.5%, P < 0.05); lymph node or distant metastasis (40.9% vs 100%, P < 0.05); or a shorter survival time after operation (50.0% vs 93.8%, P < 0.05).

CONCLUSIONSSignificant discrepancies in protein expression exist among gallbladder cancer and benign gallbladder tissues. AnnexinA3 plays an important role in the initiation and progression of human gallbladder cancer.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adult ; Aged ; Annexin A3 ; metabolism ; Carcinoma, Adenosquamous ; metabolism ; pathology ; surgery ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Cholecystitis ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; surgery ; Gene Expression Profiling ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Metastasis ; Proteome ; metabolism ; Proteomics ; Survival Rate