Objective To investigate the expression level of cyclooxygenase-2 (COX-2) and its effects on cell proliferation activity and apoptosis in colorectal carcinoma. Methods The expression of COX-2, proliferating cell nuclear antigen (PCNA) and apoptosis in 60 cases of colorectal carcinoma, 20 cases of colorectal adenoma and 20 cases of normal mucous were studied by the immunohistochemical and TUNEL methods, the apoptosis index(AI)and proliferation index(PI) were defined respectively. Results The positive expression rate of COX-2 in colorectal carcinoma was significantly higher than that in the other groups. Overexpression of COX-2 in colorectal carcinoma was related to its lymph node metastasis and the size of tumor and clinical stages. In the group with COX-2 positive expression, PI was higher than that in the group with COX-2 negative expression; whereas AI was higher in the latter group than in the former group. Conclusions The expression level of COX-2 may have a close correlation with cell proliferation and apoptosis, they may participate in oncogenesis and progression of colorectal carcinoma.

OBJECTIVETo compare the effects of Buyang Huanwu Decoction (BYHWD) and its active regions combination on brain heat shock protein 70 (HSP70) expression.

METHODSCerebral ischemia model gerbils produced by ligating bilateral common carotid artery were injected intraperitoneally by BYHWD and its active regions combination separately. HSP70 mRNA expression was determined by Northern blot test and protein expression determined by Western blot test 48 hrs after ischemia for 15 min and reperfusion.

RESULTSThe level of HSP70 mRNA and protein expression in ischemic/reperfusion brain tissue increased significantly. BYHWD and its active regions combination could decrease the HSP70 mRNA expression obviously, the inhibition effect of BYHWD was more significant than that of its active regions combination, but neither of them showed any effect on HSP70 protein expression.

CONCLUSIONBYHWD and its active regions combination might down-regulate HSP70 mRNA expression in ischemic brain tissue through its anti-ischemic effect.

Animals ; Brain ; metabolism ; Brain Ischemia ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gerbillinae ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Reperfusion Injury ; metabolism

Objective To use the fusion image of the end-inhalation holding (EIH) phase and endexhalation holding (EEH) phase to define the target volume of individual patient with liver cancer,and to evaluate the target geometry,feasibility,and clinical significance of the technology.Methods Eighteen patients with liver caucer who were treated in our hospital from 2012 to 2013 were enrolled as subjects.With the same posture and scan range,all patients underwent contrast-enhanced three-dimensional computed tomography (3DCT) scans in the phases of free breathing (FB),EIH,and EEH.Gross tumor volume (GTV),clinical target volume (CTV),and organ of risk (OAR) were delineated on the above images.CTVFB was defined as GTV on the FB phase image (GTVFB) plus a margin of 10 mm,while planning target volume (PTVFB) was defined as CTVFB plus a margin of 10 mm in the right-left and anterior-posterior directions and a margin of 20 mm in the superior-inferior direction.GTVEI and GTVEE were defined as GTV on the EIH and EEH images,respectively.Based on the EEH images,the registered EEH and EIH images were fused to form GTVEI+EI.CTVEI+EE was defined as GTVEI+EI plus a margin of 10 mm,while PTVEI+EE was defined as CTVEI+EE plus a margin of 5 mm in the right-left and anterior-posterior directions and a margin of 10 mm in the superior-inferior direction.The Pinnacle3 v8.0m treatment planning system was used to design two 3D conformal radiotherapy plans for each patient.The volume,degree of inclusion (DI),matching index (MI),and central displacement of CTVFB and CTVEI+EE,as well as PTVFB and PTVEI+EE,were compared between the two plans.Results In the 18 patients,the mean CTVFB was significantly smaller than the mean CTVEI+EE(149.00±87.54 cm3 vs.188.17± 125.72 cm3,P=0.014);there was no significant difference between the mean PTVFB and PTVEI+EE (276.68± 146.41 cm3 vs.253.66± 117.35 cm3,P=0.080).DI of CTVFB to CTVEI+EF,PTVFB to PTVEI+EE,CTVEI+EE to CTVFB,and PTVEI+EE to PTVFB were (99.83±0.09)％,(84.55±8.45) ％,(80.83± 12.31) ％,and (99.78±0.08) ％,respectively.MI of CTVEI+EE to CTVFB and PTVEI+EE to PTVFB were 0.83± 0.07 and 0.87± 0.03,respectively.The central displacements of CTVEI+EE from CTVFB in x,y,and z axes were 0.55± 1.07 cm,0.76±3.02 cm,and-0.26± 1.98 cm,respectively (P =0.432,0.971,0.587).Conclusions In the treatment of liver cancer,the target volume delineation and image fusion using 3DCT images in EIH and EEH phases may avoid target omission due to respiratory movement,making it possible to increase radiation dose to target volume and improve the efficacy of radiotherapy.

Objective]To explore the aberrant expression of SOD1 gene in nasopharyngeal carcinoma tissues and adjacent tissues,as well as in NPC cell lines,then to observe the effect of SOD1 on NPC cells metastatic ability and investigate the intrinsic?mechanism.[Methods]Immunohistochemical technique was used to examine SOD1 expression in carcinoma tissues and adjacent tissues(n=10). Small interfering RNAs and inhibitor LCS-1 were used to knockdown of SOD1 expression and inhibit SOD1 activity, respectively. Then,wound healing test and migration assay were applied to detect cell metastatic ability in vitro. Real-time PCR and Western Blot were used to analyze the expression of EMT-related genes(E-cadherin,Vimentin,Twist).[Results]SOD1 was found to be significantly up-regulated in nasopharyngeal carcinoma tissues(n = 7 ,70%),compared to control. SOD1 was also highly expressed in highly metastatic potential NPC cell lines(CNE2,5-8F,S18)compared with low metastatic ability cell lines(6-10B). Knockdown SOD1 expression or inhibit SOD1 activity suppress cell motility in CNE2 and 5-8F cells. Finally,we demonstrate that SOD1 inhibition plays a role in induction of epithelial marker E-cadherin and has an opposite effect on mesenchymal marker vimen tin and transcriptional factor twist.[Conclusion]These results suggest that SOD1 contributes to EMT and might be important for tumor metastasis in NPC.

OBJECTIVETo establish the mathematical kinetic model of the components extracted from the FTMC (formulae of the traditional Chinese medicine) and analyze parameters of the astragaloside IV extracted from the BYHWD (Buyang Huanwu decoction).

METHODThe model, including algebra and differential groups, have been set up according to the FICK discipline and Noyes-whitney soluted theories, as well as two transfer diffusive processes ((1) from protoplasate to apoplasmic, also from material compartment interior cell membrane to outside compartment; (2) apoplasmic to solution, also from outside compartment to solvent compartment) on components extraction from the FTMC. The equation groups, according to laplace transform, have been given a expression as solutions, which indicate the quantitative changes of the component concentration in solvent vs. time. The model kinetic parameters have been analyzed, meanwhile the parameters of the astragaloside IV in the BYHWD under 100 degrees C, extracted by water, have been analyzed by way of this model:

RESULTIt has been established a mathematical model that consists of three parts of e exponent. The kinetic parameters: M, alpha, N, beta, L, pi, K, k1', k2', rho1, rho2, tmax, Cmax, AUC, w0, P, D of the BYHWD were respectivelly 0.061 27% , 0.280 2 min(-1), - 1.027% , 0.008 965 min(-1), 1.077%, 0.002 665 min(-1), 3.451 x 10(-3) min(-1), 3.188 x 10(-3) min(-1), 0.375 9 min(-1), 1.420 min, 0.754 7 min, 184.9 min, 0. 0572 1 mg x mL(-1), 289.9 min, 0.070 11%, 46.24%, 22. 35%.

CONCLUSIONThe kinetic model, applied to isolated system, can have been of the rule of multiplex linear. Each parameters can be analyzed completely.

Algorithms ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Kinetics ; Mathematics ; Models, Biological ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

Objective Using twins methods to explore the influence of genetic and environmental factor on Intelligence develop-ment of children and adolescent twins .Methods It was 190 pair of twins aged 6 to 18 who lived in Chongqing were recruited ,DNA were extracted from buccal mucosa and venous blood samples to identify zygosity .The intelligence was investigated by using the Wechsler Intelligence Scale for Children(C-WISC) ,the intrapair correlation coefficients of twins were compared and calculate the genetic degrees .Results The total IQ of children aged 6 to 18 heritability were 0 .63 ,Verbal IQ were 0 .44 ,Performance IQ were 0 .57 .The total IQ heritability of children aged 10 to 14 and 15 to 18 years(0 .78 ,0 .79) were higher than that of those aged from 6 to 9 years(0 .018) .Conclusion This study suggested the intelligence of children aged 6 to 18 affected by genetic and environment , and the older ,the more influence of genetic on intelligence development .

Administration, Oral ; Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; DNA, Viral ; blood ; Female ; Guanine ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Lamivudine ; administration & dosage ; pharmacology ; therapeutic use ; Male ; Treatment Outcome ; Young Adult

OBJECTIVETo study osteoblastic phenotype expression of New Zealand rabbit skin fibroblasts transfected with mouse core binding factor a1/osteoblast specific transplanting factor-2 gene (Cbfa1/Osf2).

METHODSCbfa1/Osf2 gene, engineered into eukaryotic expression vector pSG5, was introduced into New Zealand rabbit skin fibroblasts with catholyte liposomes-Lipofectamine 2000. Meanwhile, those transfected pSG5 and un-transfected were set the control groups. The expression of Cbfa1 gene, osteocalcin (OCN) gene, alkaline phosphatase (ALP) gene and pre-peptide 2 alpha gene of collagen type I were detected by RT-PCR assay. Cbfa1 protein was detected by Western-Blot assay, in-cell ALP activity by p-nitrophenyl phosphate (PNPP) assay and OCN content in the supernatant by radio-immunity method. The ossification nodules was detected by Alizarin-Red staining and scanning electron microscope.

RESULTSCbfa 1mRNA and Cbfa1 protein were expressed in New Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 from the first day to the fifth day, but they were not detected in the control groups. In the pSG5-Cbfa1/Osf2 transfected group, the expression of ALP gene and OCN gene were respectively induced from the third day and the forth day, pre-peptide 2 alpha gene of collagen type I was enhanced from the third day. From the sixth day, ALP activity greatly increased, OCN strongly secreted, and they were maintained at a high level for about 4 weeks, and the difference was significant compared with the control group (P < 0.05). On the forty-second day, ossification nodules were found on the surface of pSG5-Cbfa1/Osf2 gene transfected cells.

CONCLUSIONSNew Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 can express osteogenesis-related genes and proteins, and form ossification nodules on their surface.

Alkaline Phosphatase ; biosynthesis ; genetics ; Animals ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; biosynthesis ; genetics ; Fibroblasts ; cytology ; physiology ; Gene Expression ; Genetic Vectors ; Mice ; Osteocalcin ; biosynthesis ; genetics ; Osteogenesis ; genetics ; physiology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo study the feasibility of osteogenic phenotype expression by human skin fibroblasts induced in polyglycolic acid (PGA) foams and the effect of tumor necrosis factor-alpha (TNF-alpha) on the expression of bone morphogenetic protein (BMP) receptors.

METHODSThe fibroblasts were isolated, purified from human skin. (1) Fibroblasts were seeded onto PGA foams. The cell-PGA complexes were cultured in RCCS for 6 weeks, in the media of TNF-alpha (50 U/ml) and BMP-2 (0.1 microg/ml). 1 d, 3 and 6 weeks later, cells and extracellular matrix were investigated by electron microscopic and histochemistry observation respectively. Secretion of osteogenic markers were analyzed by biochemical methods. (2) Fibroblasts were seeded on the glass fragments or culture flasks and treated with TNF-alpha (50 U/ml) in different usage (one-time, all-time). The RT-PCR method and the immunohistochemistry fluorescence staining were used to examine the influence of TNF-alpha on the mRNA expression and the protein expression of the type I BMP receptors at 2, 4, 6, 8 d after treatment.

RESULTSFibroblasts seeded on the PGA foams formed 3-dimensional matrix 3 weeks after seeding, which was demonstrated as osteo-tissue by tetracycline labeling and ARS staining. Cells secreted much more bone-specific alkaline phosphatase (B-AKP) and osteocalcin (OCN) into supernatant than the cells that were cultured in the media without TNF-a and BMP2. Eight days after all-time usage, the TNF-alpha (50 U/ml) increased the expression of the mRNA and protein of the type IB BMP receptor.

CONCLUSIONSFibroblasts on 3-D cell-foam structures can express osteoblastic phenotype under certain inducing conditions. The numerous fibroblasts in body would be a promising resource for cell seeds candidate of tissue- engineered bone. TNF-alpha provides the essential condition for BMP2's target effect on fibroblasts, and combined use of TNF-alpha and BMP2 is one of the regulating factors.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein Receptors, Type I ; biosynthesis ; genetics ; Bone Morphogenetic Protein Receptors, Type II ; biosynthesis ; genetics ; Bone Morphogenetic Proteins ; pharmacology ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Cells, Cultured ; Drug Synergism ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Phenotype ; Polyglycolic Acid ; Transforming Growth Factor beta ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo determine the expressions of HO-2 and eNOS in the corpus cavernosum of castrated rats and investigate the correlation of HO-2 and eNOS with erectile dysfunction.

METHODSForty SD rats of 10 weeks were equally divided into 3 castrated groups (4, 8 and 12 weeks respectively after removal of bilateral testes) and a sham operation control group. All the rats were detected for serum testosterone (T), intracavernosal pressure (ICP) and mean arterial blood pressure (MAP) 2, 8 and 12 weeks after the surgery, the content of HO-2 analyzed by Western blot and the expressions of eNOS and HO-2 determined by immunohistochemistry.

RESULTSTestosterone level and ICP/MAP ratio were markedly decreased after the castration (P < 0.05). The HO-2 expression was detected in the corpus cavernosum of all but the 12-week castrated rats, and the area under the curve was (341.50 +/- 99.70) in the 4-week castrated group, significantly less than in the 8-week (705.00 +/- 152. 74) and the control group (876 +/- 443. 36) (P < 0. 05), with no obvious difference between the latter two. eNOS was found mainly in the vascular endothelial cells of the corpus cavernosum, significantly less in the castrated (123.94 +/- 30. 23) than in the control group (421.21 +/- 125. 12).

CONCLUSIONAndrogens might mediate the penile erectile function through HO-2 and eNOS in the corpus cavernosum of rats.

Animals ; Blood Pressure ; Blotting, Western ; Erectile Dysfunction ; blood ; enzymology ; physiopathology ; Heme Oxygenase (Decyclizing) ; metabolism ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood