OBJECTIVETo study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.

METHODSImmunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.

RESULTSThe expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).

CONCLUSIONSThe decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.

Cicatrix ; metabolism ; pathology ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Serpins ; metabolism

OBJECTIVE: To analyze the expression of Maspin and Ki67 in EN-NK/TCL and the significance. METHOD: The expressions of Maspin and Ki67 were detected by immunohistochemistry in 34 cases of EN-NK/TCL and 11 cases of chronic sinusitis. RESULT: Maspin was low expressed in EN-NK/TCL, Ki67 was highly expressed in EN-NK/TCL. Negative correlation was found between the expression of Maspin and Ki67 in EN-NK/TCL. CONCLUSION The combined examination of Maspin and Ki67 has important significance in the diagnosis in EN-NK/ TCL.
Biomarkers, Tumor ; metabolism ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; Serpins ; metabolism

In order to research the bioactivity of kallistatin (Kal), we obtained the recombinant Kal using Pichia pastoris expression system. Kal cDNA was amplified from pAAV-Kal and inserted into pPIC9 vector to generate a recombinant vector of pPIC9-Kal. Then, pPIC9-Kal was linearized and transformed into Pichia pastoris strain GS115 (His4) by electroporation. The positive transformants were selected by MD plate and confirmed by PCR. High level of Kal was obtained in BMMY medium (pH 7.0) after 96 hours induction of 29 degrees C and 2% methanol, with the highest yield of 14 mg/L in shake flask culture. Kal protein was purified from the supernatant with Phenyl Superose and Heparin Sepharose FF chromatograph. The recombinant Kal had a molecular weight of 58 kDa with 98% purity, showing by SDS-PAGE. Moreover, it had a high peroxidase activity (163+/-4) U/(mgmin), which could protect LX-2 cell against oxidation of H2O2. Recombinant Kal also effectively inhibited HUVEC proliferation. In this report, we successfully established the expression system using Pichia pastoris and obtained the bioactive recombinant human Kal. It lays a foundation for its further anti-cancer therapy.
Antioxidants ; pharmacology ; DNA, Complementary ; Electroporation ; Genetic Vectors ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Serpins ; biosynthesis ; genetics

BACKGROUNDVaspin is a newly-identified adipocytokine related to obesity and insulin sensitivity. However, its pathophysiologic role in humans remains largely unknown. The aim of our study was to investigate the relationship between serum vaspin level and glucose metabolism or obesity in Chinese adults.

METHODSA total of 123 subjects, including 84 subjects with normal glucose tolerance (NGT) and 39 subjects with diabetes, were enrolled in this study. Anthropometric parameters, abdominal fat areas, plasma glucose concentration, serum insulin, lipids, and vaspin level were measured in each participant.

RESULTSSerum vaspin concentration was significantly higher in diabetic patients than that in NGT subjects (592 (438 - 695) pg/ml vs 380 (294 - 517) pg/ml, P = 0.020) in women. In all participants, age, fasting plasma glucose concentration (FPG), 2-hour post-load plasma glucose (PG2h), hemoglobin A1c (HbA1c) and high-density lipoprotein cholesterol (HDL-c) significantly increased from the lower tertile to the higher tertile of vaspin. Univariate linear regression analyses revealed that vaspin level was only positively correlated with age (beta = 0.340, P = 0.002) in NGT subjects. And vaspin was positively associated with FPG (beta = 0.365, P = 0.023), PG2h (beta = 0.526, P = 0.001), HbA1c (beta = 0.388, P = 0.016), and HDL-c (beta = 0.353, P = 0.027), while negatively with homeostasis model assessment of beta cell function (HOMA-beta) (beta = -0.361, P = 0.024) in diabetic patients. In stepwise multivariate regression analyses, age was independently associated with circulating vaspin in NGT subjects, whereas PG2h was an independent predictor of vaspin in diabetic patients. In addition, there was no significant difference of serum vaspin level between men and women. And no significant correlations between vaspin and body fat indexes were detected.

CONCLUSIONSSerum vaspin level is higher in diabetic patients than that in NGT subjects in women. Age predicts serum vaspin level in NGT subjects, while PG2h is independently associated with vaspin in diabetic patients.

Adult ; Aged ; Blood Glucose ; metabolism ; Diabetes Mellitus ; blood ; Female ; Humans ; Linear Models ; Male ; Middle Aged ; Postprandial Period ; Serpins ; blood

BACKGROUND: Maspin is a serpin family of protease inhibitors. Althouth maspin has been considered a tumor suppressor that inhibits the motility, invasion, and metastasis of breast and prostatic cancer cells, there are many conflicting reports about maspin expression and cancer prognosis. METHODS: To investigate whether the expression of maspin could be used as a prognostic marker in pancreatic cancer, 72 paraffin-embedded pancreatic ductal adenocarcinomas were analyzed using immunohistochemistry. We examined the prognostic value of maspin as well as its relationship with clinicopathological features. RESULTS: Maspin expression was observed in all pancreatic ductal adenocarcinoma. Unlike cancer tissues, however, faint or no expression was observed in the corresponding normal pancreatic tissues. In the Cox proportional hazard model, high maspin expression predicted a high hazard rate. Maspin expression had a positive correlation with tumor stage, but there were also no statistically significant relationships between maspin expression and other clinicopathological features. CONCLUSION: These findings suggest maspin expression to have biological relevance in the progression of pancreatic cancers, with potential use as a prognostic marker for pancreatic neoplasm with epithelial origin.
Adolescent ; Adult ; Aged ; Biological Markers ; Carcinoma, Pancreatic Ductal/*metabolism/surgery ; Female ; Human ; Immunohistochemistry ; Male ; Middle Aged ; Pancreatic Neoplasms/*metabolism/surgery ; Prognosis ; Proteins/*metabolism ; Serpins/*metabolism

OBJECTIVETo investigate the expression of mammary serine proteinase inhibitor(Maspin) in patients with oral squamous cell carcinoma and its relationship with clinicopathological features.

METHODSThe Maspin protein expression was examined in 45 patients with oral squamous cell carcinoma and in non-malignant oral epithelia by immunohistochemical staining. The relationship between the Maspin protein expression and the clinicopathological parameters was statistically analyzed.

RESULTSThe Maspin protein expression in the cancerous tissues of oral squamous cell carcinoma was lower than that in the non-malignant oral epithelia (P = 0.001). Negative correlation was found between the Maspin protein expression in cancerous tissue and the status of lymph node metastasis (P = 0.038). Positive Maspin protein expression indicated negative lymph node metastasis, and negative correlation was also found between the Maspin protein expression and the postoperative metastasis (P = 0.004). Positive correlation was found between the Maspin expression and the patients' survival rate (P = 0.014), patients with positive Maspin protein expression having higher survival rate than those with negative Maspin expression.

CONCLUSIONSMaspin might be useful as a potential prognostic marker for oral squamous cell carcinoma.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Serine Proteinase Inhibitors ; metabolism ; Serpins ; metabolism

OBJECTIVETo study if the expression of maspin protein in respiratory epithelial cells was downregulated through IκB kinase-α (IKKα)-controlled mechanism in an Aspergillus fumigatus-induced model in rat.

METHODSInactivated Aspergillus fumigatus hyphae (AFH) suspension was used to induce respiratory epithelial cells (REC) cultured in vitro in rat, with PBS buffer as control. By RT-PCR, the mRNA expression of maspin was quantified, and by immunocytochemistry, the expression of maspin and IKKα in REC was observed. Furthermore AFH (from level 1 to level 3) suspension was prepared to induce REC. Then Western blot hybridization technique was used to detect the expression of maspin and IKKα protein. All data were processed by analysis of variance and two-variable correlation analysis.

RESULTSBy RT-PCR, a statistically significant (t = 2.463, P < 0.05) expression of maspin mRNA was found (0.128 ± 0.059 in AFH group, 2.972 ± 0.353 in control group). By Immunocytochemistry, the difference of maspin protein color in different groups was shown statistically significant in integrated scoring (t = 3.721, P < 0.05, weak positive in AFH group, moderately positive in control group). While in IKKα color study, the difference between the two groups was also statistically significant (t = 6.825, P < 0.05) in integrated scoring, with a moderate positive in AFH group and weak positive in control group. By Western Blot hybridization, grayscale ratio of maspin and β-actin was 0.912 ± 0.023 in control group, 0.607 ± 0.030, 0.476 ± 0.019, 0.416 ± 0.017 in AFH 1-3 groups, respectively, with a statistically significant difference within the four groups (F = 281.91, P < 0.05); While grayscale ratio of IKKα and β-actin was 0.624 ± 0.012 in control group, 0.739 ± 0.020, 0.778 ± 0.010, 0.927 ± 0.017, respectively, in AFH 1-3 groups; with a statistically significant difference within the four groups(F = 200.91, P < 0.05). Moreover, the difference between any two groups from both AFH group (including subgroup 1, 2 and 3) and control group was statistically significant. Two-variable correlation analysis indicated a negative correlation between expression of maspin and IKKα (r = -0.911, P < 0.05).

CONCLUSIONSInduced by Aspergillus fumigatus, the rat respiratory epithelia might upregulate the expression of IKKα with a downregulated expression of maspin protein.

Animals ; Aspergillus fumigatus ; Cell Line ; Epithelial Cells ; metabolism ; Gene Expression Regulation ; I-kappa B Kinase ; metabolism ; Rats ; Respiratory System ; cytology ; Serpins ; metabolism

OBJECTIVE: To examine the expression of Maspin in the bladder carcinoma, and to explore the relationship among Maspin expression, clinical pathology and lymph node metastasis.
 METHODS: The expression of Maspin in 72 bladder cancer tissues and 12 normal bladder tissues were detected by immunohistochemistry. Preoperative pelvic CT images and postoperative pathological results of lymph node metastasis in 72 bladder cancer patients were analyzed retrospectively. The sensibility, specificity, positive predictive value and negative predictive value were calculated, and the advantage for diagnosis of lymph node metastasis was compared between Maspin examination and CT scan.
 RESULTS: 1) The expression of Maspin in 72 bladder cancer tissues was significantly lower than that in the normal bladder tissues (P<0.05); 2) The expression of Maspin was obviously decreased with the progress in clinical stage, pathlogical grade and lymph node metastasis, with significant difference (all P<0.05); 3) There were 27 patients diagnosed as pelvic lymph node metastasis by CT scan, and the positive rate was 37.5% (27/72); there were 22 patients diagnosed as pelvic lymph node metastasis by pathological results, and the positive rate was 30.5% (22/72). The pathological diagnose for pelvic lymph node metastasis was the gold standard. The sensitivity, specificity, positive predictive value and negative predictive value for CT was 45.5%, 73.3%, 37.0%, and 66.7%, respectively; the sensibility, specificity, positive predictive value and negative predictive value was 81.8%, 50%, 41.8% and 86.2% in the diagnosis of lymph node metastasis for the 72 bladder cancer cases by Maspin examination; the sensitivity, specificity, positive predictive value and negative predictive was 90.9%, 78.0%, 64.5% and 95.0% in the diagnose of lymph node metastasis by Maspin examination combined with CT scan.
 CONCLUSION The expression of Maspin in bladder cancer is significantly lower than that in normal bladder cancer, and a statistically significant correlation is also observed between Maspin expression and lymph node metastasis. Maspin maybe a valuable biomarker in diagnose of bladder cancer with lymph node metastasis. Maspin examination combined with CT scan has more advantage in the evaluation of bladder cancer with lymph node metastasis than Maspin or CT alone.
Biomarkers, Tumor ; metabolism ; Humans ; Immunohistochemistry ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Neoplasm Staging ; Retrospective Studies ; Sensitivity and Specificity ; Serpins ; metabolism ; Tomography, X-Ray Computed ; Urinary Bladder Neoplasms ; metabolism ; pathology

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.
Apoptosis ; Cell Proliferation ; Dependovirus ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; Serpins ; metabolism ; Signal Transduction ; Transfection

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