Adenocarcinoma, Mucinous ; enzymology ; pathology ; Cathepsin B ; metabolism ; Cathepsin D ; metabolism ; Colorectal Neoplasms ; enzymology ; pathology ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness

<b>OBJECTIVEb>To investigate the expression of cathepsin B (CatB) in colorectal cancer tissues and serum levels of CatB in patients with colorectal carcinoma and to study the association of CatB expression with lymph node and li ver metastasis.

<b>METHODSb>Immunohistochemistry was used to detect CatB expression in tissues, and enzyme linked immunosorbent assay was applied to test CatB levels in peripheral vein blood in 83 patients with colorectal cancer.

<b>RESULTSb>The expression rates of CatB in primary lesions, normal colon mucosa, lymph node metastases and hepatic metastases were 56.6%, 31.3%, 88.4%, 85.0% respectively. The positive rates of CatB in primary lesions, hepatic and lymph node metastases were higher than that in normal mucosa (chi (2)=45.6124, P< 0.01). The CatB expression rates in lymph node and hepatic metastases were higher than that in primary lesions chi (2)=11.5982, 4.3747, P< 0.05). The positive rate of CatB was higher in Dukes C and D tumors than that in Dukes A and B tumors (chi (2)=18.8871, 25.1650, P< 0.01), higher in poorly differentiated and mucous adenocarcinomas than that in well-moderately differentiated adenocarcinomas chi (2)=14.2338, P< 0.05). The mean serum level of CatB in 83 patients with colorectal cancer was (5.9+/- 2.9) ng/ml, higher than (2.3+/- 1.1) ng/ml in the controls of 30 healthy volunteers (t=6.6975, P< 0.01). The serum level of CatB in the patients with Dukes C, D stages were higher than that with Dukes A, B stages.

<b>CONCLUSIONSb>Enhanced expression of CatB in colorectal cancer tissues is associated with tumor infiltration and metastasis. Monitoring serum CatB level in patients with colorectal cancer is important in the prediction and diagnosis of lymph node and hepatic metastasis,and valuable for evaluation of the therapeutic effect.

Adult ; Aged ; Cathepsin B ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVE: To study the expression of cathepsin-B and -D in different time point after traumatic brain injury. METHODS: Traumatic brain injury (TBI) model was established on rats, cathepsin-B and cathepsin-D immunofluorescence staining and confocal microscope analysis were performed. Positive cells were counted by confocal microscope and image analysis techniques were used to determine the morphological changes in each group. RESULTS: Immunofluorescence staining results showed that cathepsin-B was activated 1 hour after TBI while cathepsin-D was not activated until 12hour after TBI. Both of them got to their peak during 4 to 8days, and kept a high level of activating 32days after TBI. Cathepsin-B and -D positive cells did not merge with caspase-3 positive cells until 6 h after TBI. CONCLUSION Cathepsin-B and -D could be the diagnostic markers of TBI and can estimating time course of lateral TBI. They blocked caspase-3 activation at the beginning period after TBI and started to promote cell death with caspase-3 6 h after TBI.
Animals ; Brain/pathology* ; Brain Injuries/pathology* ; Caspase 3/metabolism* ; Cathepsin B/metabolism* ; Cathepsin D/metabolism* ; Disease Models, Animal ; Forensic Pathology ; Hippocampus/pathology* ; Immunohistochemistry ; Lysosomes ; Male ; Neurons/metabolism* ; Rats ; Rats, Sprague-Dawley ; Time Factors

<b>OBJECTIVEb>[corrected] To investigate cathepsin B (CB) expression in squamous cervical carcinoma and its relationship to the clinical and pathological condition.

<b>METHODSb>CB expression was detected by immunohistochemistry in 56 cases of human invasive squamous cervical carcinoma (ISCC) tissues, 85 cases of cervical intraepithelial neoplasia (CIN) and 38 cases of normal cervical squamous epithelial tissue. The results were analyzed in relation to the grade of differentiation, depth of invasion and pelvic lymph node metastasis.

<b>RESULTSb>The positive rates of CB were 87.5% (49/56), 48.3% (41/85) and 48.3% (41/85) in ISCC, CIN and normal tissue, respectively. CB expression in ISCC had significant differences from that ub the CIN and normal tissues (P<0.01). CB positive rates in the tissues with invasion of less than two thirds of the cervix and over two thirds of the cervix were 83.4% (28/34) and 95.5% (21/22) respectively, showing obvious differences between them (P<0.05). CB-positive rates also showed an obvious difference between the tissues with lymphatic metastasis and those without lymphatic metastasis [97.4% (37/38) vs 66.7% (12/18), P<0.05]. CB expression in ISCC was not related to the grade of differentiation.

<b>CONCLUSIONb>High expression of CB is closely associated with tumor infiltration and lymphatic metastasis of cervical cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; Cathepsin B ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Uterine Cervical Neoplasms ; metabolism

Protein folding, stability, and function are usually influenced by pH. And free energy plays a fundamental role in analysis of such pH-dependent properties. Electrostatics-based theoretical framework using dielectric solvent continuum model and solving Poisson-Boltzmann equation numerically has been shown to be very successful in understanding the pH-dependent properties. However, in this approach the exact computation of pH-dependent free energy becomes impractical for proteins possessing more than several tens of ionizable sites (e.g. > 30), because exact evaluation of the partition function requires a summation over a vast number of possible protonation microstates. Here we present a method which computes the free energy using the average energy and the protonation probabilities of ionizable sites obtained by the well-established Monte Carlo sampling procedure. The key feature is to calculate the entropy by using the protonation probabilities. We used this method to examine a well-studied protein (lysozyme) and produced results which agree very well with the exact calculations. Applications to the optimum pH of maximal stability of proteins and protein-DNA interactions have also resulted in good agreement with experimental data. These examples recommend our method for application to the elucidation of the pH-dependent properties of proteins.
Cathepsin B ; chemistry ; metabolism ; DNA ; metabolism ; Hydrogen-Ion Concentration ; Molecular Dynamics Simulation ; Monte Carlo Method ; Muramidase ; chemistry ; metabolism ; Probability ; Protein Binding ; Proteins ; chemistry ; metabolism ; Protons ; Thermodynamics

PURPOSE: Hyperoxia has the chief biological effect of cell death. We have previously reported that cathepsin B (CB) is related to fetal alveolar type II cell (FATIIC) death and pretreatment of recombinant IL-10 (rIL-10) attenuates type II cell death during 65%-hyperoixa. In this study, we investigated what kinds of changes of CB expression are induced in FATIICs at different concentrations of hyperoxia (65%- and 85%-hyperoxia) and whether pretreatment with rIL-10 reduces the expression of CB in FATIICs during hyperoxia. MATERIALS AND METHODS: Isolated embryonic day 19 fetal rat alveolar type II cells were cultured and exposed to 65%- and 85%-hyperoxia for 12 h and 24 h. Cells in room air were used as controls. Cytotoxicity was assessed by lactate dehydrogenase (LDH) released into the supernatant. Expression of CB was analyzed by fluorescence-based assay upon cell lysis and western blotting, and LDH-release was re-analyzed after preincubation of cathepsin B-inhibitor (CBI). IL-10 production was analyzed by ELISA, and LDH-release was re-assessed after preincubation with rIL-10 and CB expression was re-analyzed by western blotting and real-time PCR. RESULTS: LDH-release and CB expression in FATIICs were enhanced significantly in an oxygen-concentration-dependent manner during hyperoxia, whereas caspase-3 was not activated. Preincubation of FATIICs with CBI significantly reduced LDH-release during hyperoxia. IL-10-release decreased in an oxygen-concentration-dependent fashion, and preincubation of the cells with rIL-10 significantly reduced cellular necrosis and expression of CB in FATIICs which were exposed to 65%- and 85%-hyperoxia. CONCLUSION: Our study suggests that CB is enhanced in an oxygen-concentration-dependent manner, and IL-10 has an inhibitory effect on CB expression in FATIICs during hyperoxia.
Animals ; Cathepsin B/*genetics/metabolism ; *Down-Regulation ; Gene Expression Regulation ; Hyperoxia/*genetics ; Interleukin-10/*pharmacology/physiology ; L-Lactate Dehydrogenase/metabolism ; Necrosis/chemically induced ; Oxygen/metabolism ; Rats

Adult ; Aged ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Cathepsin B ; biosynthesis ; genetics ; Female ; Humans ; Lung Neoplasms ; metabolism ; Male ; Middle Aged

Biomarkers, Tumor ; metabolism ; Carcinoma, Papillary ; metabolism ; Cathepsin B ; metabolism ; Cell Cycle Proteins ; metabolism ; HSP27 Heat-Shock Proteins ; metabolism ; Humans ; Proteomics ; Repressor Proteins ; metabolism ; S100 Calcium Binding Protein A6 ; S100 Proteins ; metabolism ; Serpins ; metabolism ; Thyroid Neoplasms ; metabolism

<b>OBJECTIVESb>To detect the expression of Cathepsin B (CatB) in the intracranial aneurysm wall and its effect to the apoptosis of vascular smooth muscle cells, aimed at clarifying the pathological formation mechanism of intracranial aneurysm.

<b>METHODSb>From November 2006 to February 2009, 20 intracranial aneurysm samples were collected as the experimental group, and 6 cases of normal pallium artery samples were collected as the control group. Immunohistochemical technique was used to evaluate the expressions of CatB, alpha-smooth muscle actin (alpha-SMA) and Caspase-3. The expression of CatB mRNA was evaluated by real-time PCR. The ultrastructure of intracranial aneurysms were observed by using the transmission electronic microscope.

<b>RESULTSb>Compared with the normal pallium artery specimens, the expression of CatB and Caspase-3 both significantly increased in the intracranial aneurysm walls where alpha-SMA decreased (P < 0.05). The mean expression of CatB mRNA in intracranial aneurysm samples was about 3.8-folds than that in control group (P < 0.01). There were excessive apoptotic vascular smooth muscle cells (VSMCs) in the tunica median, and typical apoptotic body were observed in some aneurysm walls.

<b>CONCLUSIONb>Cathepsin B may be involved in the formation and the progression of intracranial aneurysm.

Adolescent ; Adult ; Aged ; Apoptosis ; Case-Control Studies ; Cathepsin B ; metabolism ; Female ; Humans ; Intracranial Aneurysm ; enzymology ; pathology ; Male ; Middle Aged ; Muscle, Smooth, Vascular ; pathology ; Young Adult

<b>OBJECTIVEb>To construct the yeast two-hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma.

<b>METHODSb>We have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma.

<b>RESULTSb>We have screened several proteins that interaction with FasL protein, including metallothionein 1K, 1G, 2A, cathepsin B, fatty acid synthase, interferon alpha-inducible protein 27, phospholipid scramblase, Ser/Thr-like kinase, anchor attachment protein, fibulin-5.

<b>CONCLUSIONSb>We have successfully constructed the yeast two-hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.

Cathepsin B ; metabolism ; Cloning, Molecular ; Colorectal Neoplasms ; chemistry ; pathology ; Fas Ligand Protein ; Gene Library ; Humans ; In Vitro Techniques ; Liver Neoplasms ; chemistry ; secondary ; Membrane Glycoproteins ; genetics ; metabolism ; Metallothionein ; metabolism ; Protein Binding ; Tumor Necrosis Factors ; genetics ; metabolism ; Two-Hybrid System Techniques ; Yeasts ; genetics