Breast Neoplasms ; metabolism ; pathology ; Female ; Humans ; Insulin-Like Growth Factor Binding Protein 5 ; biosynthesis ; genetics ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured

Objective To evaluate operative efficacy in regional pancreaticoduodenectomy with mesenteric-portal vein resection and graft reconstruction using iuternal jugular vein for pancreatic head carcinoma.Methods From Jan 2000 to Jan 2003,6 patients with pancreatic head tumors and mesenteric- portal vein involvement underwent regional pancreatieoduodenectomy with mesenteric-portal vein reseetion and vascular reconstruction using internal jugular vein.Results Surgery was successful in all 6 patients. Postoperative pathology revealed that mesenteric vein or portal vein were invaded by tumor in all cases. Survival time ranged from 17 to 49 mouths.The median survival was 23.4 months.Two cases have survived over 3 years and one of them was alive 49 months postoperatively without recurrence.Conclusion The regional pancreaticoduodenectomy with tumor invaded mesenteric-portal vein resection and graft reconstruction by using internal jugular vein renders a longer survival in metastasis-negative patients of pancreatic head adenocarcinoma.

Breast Neoplasms ; genetics ; metabolism ; Female ; Gene Expression Profiling ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; methods

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Objective To explore the mechanism of protecting cells from hypoxia/ reoxygenation (H/R) injury by constructing specific small interference RNA (siRNA) to inhibit NALP3 expression in rat renal tubular epithelial cells (NRK-52E).Methods (1) To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 1 h,2 h,4 h,8 h,16 h and 24 h.The activity of lactae dehydrogenase (LDH) in the culture medium,cell count and cell viability,the expression of NALP3 were determined by biochemical method,trypan blue exclusion and Western blotting.(2) The siRNA was transfected into NRK-52E.The irrespective siRNA transfected group wasused as control.NALP3 expression was examined by Western blotting.(3) The cells were divided into 4 groups:control group,H/R group,irrespective siRNA transfected group and NALP3-siRNA transfected group.To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 4 h.And the expression of NALP3 was determined by Western blotting.(4)Cellular apoptosis was examined by Annexin V/PI staining and flow cytometry.NF-κB DNA binding activity,IκB-α,Bcl-2 and Bax expression were examined by EMSA and Western blotting.Results (1)Compared with the control group,the activity of LDH significantly increased,cell count and cell viability significantly decreased (all P＜0.05).The expression of NALP3 significantly increased and peaked at 4 h after H/R.(2)The specific siRNA could efficiently inhibit NALP3 expression in NRK-52E.Compared with the irrespective siRNA transfected group,the protein expression of NALP3 was significantly down-regulated in NALP3 siRNA transfected group (P＜0.05).(3)After hypoxia 1 h and reoxygenation 4 h,the activity of LDH and the expression of NALP3 increased.Compared with the irrespective siRNA transfected group,LDH concentration in media and the expression of NALP3 significantly decreased in NALP3-siRNA transfected group.(4)After hypoxia 1 h and reoxygenation 4 h,NF-κB DNA binding activity was increased,IκB-α phosphorylation and degradation,Bcl-2 and Bax were significantly up-regulated.However,compared with the irrespective siRNA transfected group,NF -κB DNA binding activity,IκB-α degradation and Bax/Bcl-2 were significantly decreased (P＜0.05) in NALP3-siRNA transfected group.At the same time,the ratio of apoptosis was significantly increased in three groups than that in control.Compared with the irrespective siRNA transfected group,the ratio of apoptosis in NALP3-siRNA transfected group was significantly decreased (P＜0.05).Conclusions H/R induces the expression of NALP3 in NRK-52E.The synthesized siRNA can inhibit the expression of NALP3 and protect NRK-52E from hypoxia/reoxygenation injury.The mechanism may be via inhibiting the activation of NF-κB,modulating expression of Bcl-2 and Bax,as well as decreasing cell apoptosis.

OBJECTIVETo screen, clone and identify the cDNA fragments of human endometrial carcinoma-related genes,and explore the molecular mechanism of endometrial carcinogenesis.

METHODSPure endometrial glandular epithelial cells and endometrial carcinoma cells were obtained by laser capture microdissection (LCM). RNA from these cells was isolated, and differentially expressed gene fragments that were specialy relevant to endometrial carcingenesis were identified by using fluorescence differential display reverse transcription polymerase chain reaction (FDD-PCR). The selected fragments were cloned, sequenced and verified by reverse Northern blot analysis, and positive fragments were BLAST analysed and compared with those in Genbank.

RESULTS38 differential fragments were isolated, 3 of which were expressed more abundantly in normal endometrium and 35 were highly expressed in endometrial carcinoma. 10 fragments were recoverd, cloned and sequenced, confirmed by reverse Northern blot analysis, among which 6 fragments were positive. BLAST analysis showed that T1.1 was homologous to cyclin-dependent protein kinase 7 (CDK7, 99%); L1.9 was homologous to protein phosphatase 1 regulatory (inhibitor) subunit 12A (PPP1R12A, 99%); L1.21 and L1.22 were homologous to cellular repressor of E1A-stimulated genes 1 (CREG, 100%); L1.25 and L1.26 were homologous to solute carrier family 39 (zinc transporter) member 10 (SLC39A10, >98%).

CONCLUSIONGene fragments related to endometrial carcinoma have been obtained by applying LCM and FDD-PCR. To our knowledge it is the first time that the correlation between CDK7, PPP1R12A, CREG, SLC39A10 and endometrial carcinoma is discovered at mRNA level, and their role in molecular mechanism of cancinogenesis is discussed. CDK7, CREG, SLC39A10 as new candidate oncogene and PPP1R12A as new candidate anti-oncogene are worthy of being further investigated in the future.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Cation Transport Proteins ; genetics ; metabolism ; Cyclin-Dependent Kinases ; genetics ; metabolism ; DNA, Complementary ; genetics ; Endometrial Neoplasms ; genetics ; metabolism ; pathology ; Female ; Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; Myosin-Light-Chain Phosphatase ; genetics ; metabolism ; Oncogenes ; RNA, Messenger ; metabolism ; RNA, Neoplasm ; genetics ; Repressor Proteins ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo study the methods of diagnosis and treatment of malignant peritoneal mesothelioma.

METHODSThe clinical data of 41 patients with malignant peritoneal mesothelioma pathologically confirmed were retrospectively analyzed.

RESULTSOf these 41 patients, abdominal pain and diarrhea were found in 38 and 35. The histopathologic types were: epitheloid, fusiform and mixed in 21, 11 and 9 cases. CT was performed in 21 and laparoscopic exploration in 13. The overall surgical resection rate was 70.7%. After operation, local abdominal cavity chemotherapeutic instillation was given in 12, systemic chemotherapy in 23 and immunotherapy in 3. The two year survival rate was 36.6%.

CONCLUSIONFor malignant peritoneal mesothelioma, the final diagnosis depends on histological examination and the treatment should be surgical-core combined methods.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Cisplatin ; therapeutic use ; Combined Modality Therapy ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Mesothelioma ; diagnosis ; drug therapy ; surgery ; Middle Aged ; Peritoneal Neoplasms ; diagnosis ; drug therapy ; surgery ; Retrospective Studies ; Survival Rate ; Tomography, X-Ray Computed

Patients with pulmonary metastasis from colorectal cancer have been considered to be associated with poor prognosis. It is a problem to improve survival for patients who suffer pulmonary metastasis from colorectal cancer by analyzing the prognosis of patients who underwent pulmonary surgery or not and then choose the right treatment regimen. The identification of prognostic factors is particularly important in colorectal cancer patients with pulmonary metastasis.
Colorectal Neoplasms ; diagnosis ; therapy ; Humans ; Lung Neoplasms ; diagnosis ; therapy ; Prognosis

OBJECTIVETo study the relationship between collgen (Col) IV, VEGF secreted by the tumor cells and vasculogenic mimicry (VM).

METHODS158 bi-phase differential malignant tumor specimens were alloted and made into tissue microarray. These tissue microarray sections were stained with CD31, periodic acid-Schiff (PAS) and Col IV. Subsequently, distributive trait of Col IV and the difference of VEGF expression were analyzed.

RESULTSThe basement membrane of VM was PAS and Col IV positive. The expression of VEGF in bi-phase differential malignant tumor with VM was less than that in those without VM. The difference of VEGF expression in malignant melanoma and alveolar rhabdomyosarcoma was significant (P < 0.05).

CONCLUSIONCollgen IV and periodic acid-Schiff positive material take part in constructing the basement membrane of vasculogenic mimicry. The difference of the VEGF expression proves that vasculogenic mimicry can sustain the tumor blood supply.

Collagen Type IV ; analysis ; Humans ; Immunohistochemistry ; Neoplasms ; blood supply ; chemistry ; pathology ; Periodic Acid-Schiff Reaction ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo explore the potential mechanisms of leukemia cell resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) -induced apoptosis.

METHODSCells apoptosis, changes of mitochondrial membrane potential, activity of NF-kappaB, activity of caspase-8 and expressions of apoptosis-related proteins in TRAIL treated K562 and CEM cells, were detected by flowcytometry, ELISA and Western blotting methods, respectively.

RESULTSAfter treated with TRAIL, the apoptosis indexes were 29.98% and 14.1%, and mitochondrial membrane potential were decreased to 73.25% and 25.4% in K562 and CEM cells respectively. Constitutive level of caspase-8 expression in CEM was lower than that in K562 cells. Both cells became over-expressed Bcl-xL and down-regulated Bax. The ratio of Bcl-xL/Bax in CEM cells was higher than that in K562 cells. Compared with that in K562, the NF-kappaB activity increased significantly in CEM after treatment with TRAIL in early stage.

CONCLUSIONCEM cells were more resistant to TRAIL-induced apoptosis than K562 cells did. The potential mechanisms associated with CEM drug resistance might be the lower expression of the constitutive level of caspase-8, lower sensitivity of mitochondrial inner membrane, early increase in NF-KB activity and altered expression of Bcl-2 proteins family.

Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 8 ; metabolism ; Drug Resistance, Neoplasm ; Humans ; K562 Cells ; Ligands ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Tumor Cells, Cultured