OBJECTIVETo explore the clinicopathological features, immunophenotype, differential diagnosis, pathogenesis and prognosis of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract.

METHODSClinical and pathologic findings of 3 cases of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract were analyzed by gross examination, microscopic investigation and immunohistochemical staining. The related literatures were reviewed.

RESULTSAll of the three cases were middle-aged or elderly patients. Three cases all presented with hematuria and mucusuria. Endoscopic examination identified that case 1 had a polyp with broad attachment in the dome of bladder, case 2 had a solid mass in the ureter, and case 3 had a exophytic fungating tumor in the renal pelvis. Microscopically, case 1 revealed a papillary lesion with finger-like processes lined by pseudostratified columnar epithelium with abundant goblet cells. The cells demonstrated moderate degree dysplasia. In case 2 and case 3, both villous adenomas and poorly differentiated adenocarcinoma were observed, the adenoma cells arranged in a cribriform pattern, and the tumor cells showed severe atypia, mitotic activity, and transition with invasive poorly differentiated adenocarcinoma. Immunohistochemically, the tumor cells in three cases were positive for CK20, CEA,EMA and MUC-1; none of them expressed cdx-2 and PSA; In case 2 and 3, the same immunophenotype of villous adenomas and their associated adenocarcinomas was observed, but the number of the positive cells of p53 and Ki-67 staining were significantly increased in the area of adenocarcinomas than in that of the villous adenomas.

CONCLUSIONSVillous adenoma of the urinary tract is rare. It can occur in the urinary bladder, urachus, renal pelvis, ureter and urethra. These lesions may have malignant potential and frequently coexist with other malignant tumors. So, villous adenoma of the urinary tract should be removed completely and sampled thoroughly to avoid missing a more aggressive component.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenoma, Villous ; metabolism ; pathology ; secondary ; surgery ; Adult ; Aged ; Carcinoembryonic Antigen ; metabolism ; Follow-Up Studies ; Humans ; Keratin-20 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Kidney Pelvis ; Lung Neoplasms ; secondary ; Male ; Mucin-1 ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Ureteral Neoplasms ; metabolism ; pathology ; surgery ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery

Objective To investigate the curative effect of acute severe brain injury complicated ARDS, Methods 31 patients who had acute severe brain injury complicated ARDS were divided into two groups:A group was early discovery of ARDS and given treatment.B group was late discovery of ARDS and treated late.Then the curative effects were compared.Results A group was significantly higher than B group in blood gas analysis(P

OBJECTIVETo compare the durability of quantum dots with that of green fluorescein for labeling survivin antisense oligonucleotide (ASODN) and investigate the difference in growth and apoptosis of cells transfected with the labeled survivin ASODN.

METHODSSurvivin ASODN labeled with quantum dots or green fluorescein was transfected into MCF-7 cells via Lipolifectmain(TM2000). The proliferation of MCF-7 cells was assessed with MTT assay, survivin mRNA expression determined by RT-PCR and its protein expression measured by Western blot analysis. The apoptosis rate of the transfected cells was estimated by flow cytometry, and the fluorescence distribution in the cells observed under fluorescent inverted microscope.

RESULTSThe mRNA and protein expressions of survivin were significantly decreased in the MCF-7 cells after cell transfection with survivin ASODN labeled with quantum dots or green fluorescein, and no significant difference was noted between the two labeling methods (P>0.05). Nor did survivin ASODN transfection with different labeling methods produced significant difference in cell proliferation and apoptotic rate (P>0.05). For green fiuorescein labeling, the fluorescence disappeared 4 days after transfection, whereas the fluorescence sustained for 1 week for quantum dots labeling.

CONCLUSIONSurvivin ASODNs labeled with quantum dots and green fiuorescein do not significantly differ in survivin expression or the transfected cell proliferation and apoptosis rate, but quantum dot labeling can be more stable with longer maintcnance of the labeling.

Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Fluorescein ; chemistry ; Gene Expression ; Humans ; Inhibitor of Apoptosis Proteins ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; chemistry ; genetics ; Quantum Dots ; Reverse Transcriptase Polymerase Chain Reaction ; Staining and Labeling ; methods ; Transfection

OBJECTIVETo evaluate the specific killing effects of combination of recombinant adenovirus mediated double suicide gene driven by KDR promoter and survivin antisense oligonucleotide(ASODN) on colorectal cancer cells and vascular endothelial cells.

METHODSThe 293 packaging cells were transfected with the plasmids of pAdEasy-CDglyTK and the recombinant adenovirus were generated. The KDR expressive cells of SW620, ECV304 were infected with adenovirus, meanwhile survivinASODN was transferred into the same cells. The infection rate of adenovirus and transfection efficiency of survivinASODN were observed and the expression of CDglyTK was detected by RT-PCR. The expression of survivin was measured by Western blot. The killing effects and bystander effects on SW620, ECV304 were examined through MTT method.

RESULTSThe cells which were infected with the adenovirus mediated double suicide gene could be transfected with the survivin ASODN and the infection rate was not affected as well as the transfection efficiency. The high expression of CDglyTK gene was found in SW620, ECV304 cells infected with recombinant adenovirus and survivin ASODN decreased the survivin protein level. The survival rate of gene therapy group was significantly lower than that of negative group. The combination of survivin ASODN and AdKDR-CDglyTK gene therapy showed significantly lower survival rate of SW620 and ECV304 cells as compared with the AdKDR-CDglyTK or survivin ASODN used alone (P<0.05). The survival rate was slightly lower in GCV 100 microg/ml, 5-FC 2000 microg/ml than that AdKDR-CDglyTK used alone (P>0.05). The combined therapy of AdKDR-CDglyTK and survivin ASODN showed synergistic killing efficacy and more significant bystander effects.

CONCLUSIONThe combined gene therapy of AdKDR-CDglyTK system and survivin ASODN has stronger specific killing effects on colorectal cancer cells and vein endothelial cells.

Adenoviridae ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; Endothelial Cells ; metabolism ; Genes, Transgenic, Suicide ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Receptors, Vascular Endothelial Growth Factor ; genetics ; Transcription Initiation Site

BACKGROUNDThis study was undertaken to obtain differentially expressed genes related to human glioma by cDNA microarray and the characterization of a novel full-length gene.

METHODSTotal RNA was extracted form human glioma and normal brain tissue, and mRNA was used as a probe. The results of hybridization procedure were scanned with the computer system. The gene named 507E08 cone was subsequently analyzed by northern blot, bioinformatic approach, and protein expression.

RESULTSFifteen differentially expressed genes were obtained from human glioma by hybridization and scanning for four times. Northern blot analysis confirmed that the 507E08 clone was low expressed in human brain tissue and over expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that the 507E08 clone was a novel full-length gene, which codes 203 amino acid of protein and is called human ribosomal protein 14.22 gene. The nucleotide sequence had been submitted to the GenBank with the accession number of AF329277. After expression in E. coli., protein yielded a major band of apparent molecular mass 22 kDa on an SDS-PAGE gel.

CONCLUSIONScDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human ribosomal protein 13.22 may be correlated with the development of human glioma.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Glioma ; genetics ; pathology ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism ; Ribosomal Proteins ; genetics ; metabolism ; Sequence Analysis, DNA

Objective To evaluate the effect of long-term microwave radiation on the expression of N-methyl-D-aspartate receptor(NMDAR),brain derived neurotrophic factor(BDNF) and related molecules in signal pathways in the hippocampus of rats.Methods Fifty male Wistar rats were exposed to microwave radiation at an average power density of 0,5,10,20 and 30 mW/cm2for 6 min/time,3 times/week,and for 6 weeks,which were sacrificed and the hippocampus was quickly removed at 14 d and 28 d after exposure.The changes in NMDAR (NR1,NR2A,NR2B),postsynaptic density protein(PSD)-95,cortactin,BDNF and tyrosine kinase receptor B (TrkB) in hippocampal neurons of each group were detected by Western blotting and image analysis techniques.Results Compared with the control group,the expressions of related proteins did not change significantly after microwave irradiation of 5 mW/cm2 at each time point.After 20 mW/cm2 microwave radiation,the expression of NR1 was increased at 14 and 28 d (P ＜0.05),the expression of NR2A was increased at 28 d (P ＜ 0.05),but the expression of NR2B was decreased at 14 and 28 d (P ＜ 0.05).At a average power density of 30 mW/cm2,the expressions of NR1,NR2A and PSD-95 and the expression of NR2B were decreased at 14 and 28 d(P ＜0.05),and cortactin,BDNF and TrkB were increased at 14 d after irradiation (P ＜ 0.05).Conclusion The effect of different dosages of long-term microwave radiation on the proteins of NMDAR and its signal pathway related molecules is different.Microwave radiation may affect the NMDAR of postsynaptic information transmission through the BDNF-TrkB signaling pathways,which might play an important role in the impediment of learning and memory function caused by microwave radiation.

OBJECTIVETo investigate the effect of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy (PDT) on MGC-803 human gastric cancer cells in vitro.

METHODSMGC-803 human gastric cancer cells were treated with 5-ALA at various concentrations followed by laser irradiation. The cells were also treated with 5-ALA at the same concentration before laser exposure at various doses. PDT-induced phototoxicity of the cells was determined by MTT assay.

RESULTSAfter laser exposure of the cells at the same dose (25.0 J/cm(2)), the cell survival rates decreased significantly with incubation of the cells with 5-ALA at 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L, respectively (F=266.39, P<0.001), but 2.0 and 4.0 mmol/L ALA showed no significant difference in lowering the cell survival rates (P>0.05). Following treatment with the same 5-ALA concentration (1 mmol/L), the cell survival rates decreased in response to increased laser doses (at 6.25, 12.5, 25.0, 50.0, and 100 J/cm(2), respectively, F=226.31, P<0.0001). Without laser exposure, the survival rate of the cells did not significantly change for different 5-ALA concentrations (F=0.79, P=0.5383), nor did it undergo obvious variation in response to different laser doses without 5-ALA incubation (F=0.61, P=0.6551).

CONCLUSIONSThe damage of MGC-803 cells by PDT increases with 5-ALA concentration within a relative lower range and is proportional to the laser doses delivered. Without 5-ALA treatment, the laser at the chosen dose cannot produce photodynamic effect and ALA itself is nontoxic. ALA-mediated PDT appears to be a promising therapy for gastric cancer.

Aminolevulinic Acid ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Dose-Response Relationship, Drug ; Dose-Response Relationship, Radiation ; Humans ; Lasers ; Photochemotherapy ; Photosensitizing Agents ; pharmacology ; Stomach Neoplasms ; drug therapy ; pathology

OBJECTIVETo evaluate the curative effect of curved cutter stapler (Contour, Ethicon Endo-Surgery, Inc) in the ultra low anterior resection for low rectal cancer.

METHODSClinic data of 56 patients with low rectal cancer from Dec. 2005 to Sep. 2006 were reviewed retrospectively. After total mesorectal excision (TME) and lateral lymph node dissection (LLD) in 56 cases, the rectal (anal) remnant was cut and closed with curved cutter stapler (Contour), and preserved for ultra low colo-rectal (anal) anastomoses with 33 mm straight intraluminal stapler.

RESULTSThere was no operational death and the mean hospitalization time was (11.2+/-3.2) days. The incidence rate of postoperative complications in 1 month was 3.57% (2/65). Both of the cases were anastomotic leakage. One was cured by surgical drainage, the other combining with rectal vaginal fistula was cured by transverse colostomy.

CONCLUSIONCurved cutter stapler has the advantages of complete cutting, safe closure and low anastomotic leakage rate in the process of ultra low anterior resection for low rectal cancer.

Adult ; Aged ; Anastomosis, Surgical ; instrumentation ; methods ; Female ; Humans ; Middle Aged ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; pathology ; Retrospective Studies ; Surgical Equipment

BACKGROUNDIn patients suffering from acute pancreatitis, the pathogenesis is not completely understood, and several recent studies in vitro suggested that heat shock proteins might play an important role in cell signaling. To investigate the possible role of extracellular heat shock protein 70 (Hsp70) in pancreatitis, toll-like receptor-4 (TLR4)-deficient and wild-type mice were administered with exogenous Hsp70 during the course of cerulein-induced pancreatitis (CIP).

METHODSAcute pancreatitis was induced by 5 intraperitoneal injections of cerulein at hourly intervals, and then treated with recombinant Hsp70 through the caudal vein 4 hours after the start of cerulein injections. Subsequently serum amylase and serum cytokines levels were detected. Histologic alteration of the pancreas was evaluated. Tumor necrosis factor alpha (TNF-alpha) concentrations and myeloperoxidase (MPO) activity in both pancreas and lungs were analyzed. The nuclear factor kappa B (NF-kappaB) activation in pancreatic tissue was measured using a sensitive RelA enzyme-linked immunosorbent assay.

RESULTSTreatment with recombinant Hsp70 to wild-type mice in CIP resulted in significant aggravation of inflammation in pancreas, elevated levels of serum cytokines, up-regulation of pulmonary MPO activity and increase of lung tissues TNF-alpha concentrations. In contrast, treatment with Hsp70 to TLR4-deficient mice had little effect on serum cytokines levels, pancreatic inflammation, pulmonary MPO activity and TNF-alpha concentrations.

CONCLUSIONSThe results suggest that extracellular Hsp70 might induce systemic inflammatory response syndrome (SIRS)-like response in vivo and TLR4 might be involved in the Hsp70-mediated activation of inflammatory reaction in the progression of CIP without infection.

Acute Disease ; Animals ; Ceruletide ; toxicity ; Female ; HSP70 Heat-Shock Proteins ; physiology ; Male ; Mice ; Mice, Inbred C57BL ; Pancreatitis ; etiology ; Systemic Inflammatory Response Syndrome ; etiology ; Toll-Like Receptor 4 ; physiology

OBJECTIVETo establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells.

METHODSWe cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31(+) cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay.

RESULTSThis approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel.

CONCLUSIONSThe improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.

Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Line ; Collagen Type I ; Culture Media ; Embryonic Stem Cells ; cytology ; Endothelial Cells ; cytology ; Humans