The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Fibroblasts ; cytology ; physiology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Mice ; Peptides ; chemistry ; Water ; chemistry

OBJECTIVETo evaluate the mechanism of increased invasion and migration of hepatocellular carcinoma (HCC) cells induced by vascular endothelial growth factor receptor-1 (VEGFR-1) activation.

METHODSVascular endothelial growth factor-B (VEGF-B) was used to induce and stimulate hepatocellular carcinoma cell line MHCC97-H. Morphologic changes of MHCC97-H were investigated. The expression of E-cadherin and alpha-catenin (two epithelial markers) and Vimentin and N-cadherin (two mesenchymal markers) was detected by reverse transcriptase polymerase chain reaction (RT- PCR), western blotting, and immunofluorescence staining. Cell invasion and migration test was performed.

RESULTSTreatment of MHCC97-H cells with VEGF-B led to morphologic changes characteristic of epithelial to mesenchymal transition (EMT), including loss of polarity, increased intercellular separation, and the presence of pseudopodia. Expression of the epithelial adhesion molecules, including E-cadherin and alpha-catenin, was decreased after VEGF-B treatment. Conversely, an increase in the expression of the mesenchymal cell markers, including N-cadherin and vimentin, was observed after VEGF-B treatment (P less than 0.05). VEGF-B-treated cells exhibited a change in E-cadherin from an organized, membrane-bound structure to a disorganized state in which it was noted to be dispersed throughout the cytoplasm. Pretreatment with VEGFR-1 blocking antibody 18F1 inhibited the change in localization of E-cadherin induced by VEGF-B treatment. The ability of invasion and migration of MHCC97-H was enhanced by VEGF-B reatment (P less alpha 0.05).

CONCLUSIONIncreased invasion and migration of HCC cells induced by VEGFR-1 activation was mediated by epithelial to mesenchymal transition.

Carcinoma, Hepatocellular ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Liver Neoplasms ; Vascular Endothelial Growth Factor A ; metabolism

Objective To investigate the effect of fibronectin type Ⅲ and SPRY domain containing 1 (FSD1) protein on the invasion of glioma stem cells (GSCs), so as to probe into the new biomarkers or potential therapeutic targets for gliomas. Methods The Cancer Genome Altas (TCGA) database data were used to analyze and compare the FSD1 gene expression (the FSD1 mRNA level) in the glioblatoma (also known as glioblastoma multiforme, GBM) and normal brain tissues as well as in the different grade glioma tissues, and the correlation of the FDS1 gene expression (mRNA level) with the survival prognosis of patients was also analyzed using the TCGA database data. The lentivirus was used to overexpress the FSD1 protein in the GSCs, T4121 and D456. The effect of the overexpressed FSD1 protein on the invasive ability of the GSCs, T4121 and D456 was evaluated by Transwell invasion assay. Results The FSD1 gene expression (mRNA level) was significantly lower in GBM than in normal brain (P＜0.01). The FSD1 gene expression (the mRNA level) in gliomas significantly decreased with the increase of the gliomas grade (gradeⅡvs Ⅲ, P＜0.05;gradeⅢvs Ⅳ, P＜0.01). The survival prognosis of patients with gilomas was well associated with the level of FSD1 gene expression (the FSD1 mRNA level), as indicated by the overall survival rate of the patients, which was significantly lower in the patients with the low FSD1 mRNA level than in the patients with the high FSD1 mRNA level (P＜0.01). In the Transwell invasion assay, the count of the invasive cell numbers significantly decreased in the FSD1 protein-overexpressed T421 and D456 groups than in the corresponding control group (P＜0.01 in both T4121 and D456 cell lines). Conclusion There is a clinical relevance of the FSD1 expression for the malignant progression of gliomas (the grade of gliomas). The low level FSD1 is favorable for keeping the invasive ability in GSCs.

OBJECTIVETo study the effects of Liangge San to the expression of CD14 and scaverger receptor(SR) in the kupffer cells of liver and the pathological changes of liver tissue of endotoxemia-mice.

METHODThe model was established with intravenous injection of lipopolysaccharide (LPS) and at the same time different dose Liangge San were given. The expression of CD14 and scaverger receptor were detected with immunohigtochemistry at the 2nd, 4th, 8th hour ofter injury and analyzed with computer image system, and the pathological changes of liver tissue were also observed.

RESULTAt the three different hours, the expression of CD14 and scaverger receptor in macrophages of liver of LPS-injury group showed significant increase and significant decrease respectively, compared with that of the blank-control group (P < 0.01). The expression in dexamethasone group and Liangge San different dose groups were intermediate between those in injury group and those in control group. Compared with expression of LPS-injury group, those of dexamethasone group and Liangge San different dose groups showed significant differences (P < 0.01), especially that of Liangge San high dose group. Liver cells showed vacuole change. Changes of CD14 and SR expression were paralleled with the severity of liver damages of the mice.

CONCLUSIONLiangge San can inhibite the up-regulation of CD14 expression and down-regulation of scaverger receptor expression in a dosage-dependent manner and also alleviate the damages of liver induced by LPS.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endotoxemia ; chemically induced ; metabolism ; pathology ; Female ; Kupffer Cells ; metabolism ; pathology ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; Liver ; metabolism ; pathology ; Mice ; Plants, Medicinal ; chemistry ; Receptors, Scavenger ; metabolism

OBJECTIVEThe sex therapy is not yet popularized at present. This study aimed to evaluate the effect of the combination of the improved sex therapy and oral sildenafil on erectile dysfunction (ED).

METHODSA total of 3130 Uigur cases of ED received in Xinjiang Bogda Hospital were divided into a control group (n=625) and a trial group (n=2505), the former treated with oral sildenafil alone, and the latter by the combination of the improved genital therapy and sildenafil, both for 3 months and followed up at 6 and 12 months after the treatment. The therapeutic effects were evaluated and compared using IIEF-5.

RESULTSThe IIEF-5 scores of the control group were 12.80 +/- 3.76 and 18.10 +/- 2.61 before and after the treatment, and 17.35 +/- 2.73 and 16.64 +/- 2.63 at 6 and 12 months, respectively, while those of the trial group were 12.73 +/- 3.52 and 19.06 +/- 4.07 before and af- ter the treatment, and 19.86 +/- 2.42 and 20.47 +/- 2.38 at 6 and 12 months, respectively, with statistically significant differences either between pre- and post-treatment (P < 0.05) or between the control and trial groups at 6 and 12 months (P < 0.05).

CONCLUSIONThe combination of the improved sex therapy and oral sildenafil is superior to sildenafil alone in the treatment of ED, and its efficacy is relatively stable at 12 months.

Adult ; Aged ; Asian Continental Ancestry Group ; Erectile Dysfunction ; drug therapy ; ethnology ; Humans ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Purines ; therapeutic use ; Retrospective Studies ; Sildenafil Citrate ; Sulfones ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the efficacy, median time to progression (TTP), quality of life and toxicity in the patients with advanced non-small cell lung cancer (NSCLC), treated with thalidomide plus vinorelbine and cisplatin (NP) or NP alone.

METHODSSixty six patients with advanced NSCLC were divided randomly into two groups, the trial and control groups. The trial group was treated with vinorelbine 25 approximately 30 mg/m(2) i.v. on D1 and D8, cisplatin 70 approximately 80 mg/m(2) i.v. on D1 (NP regimen), and thalidomide 200 mg orally and daily from D1. The control group received vinorelbine and cisplatin as above described.

RESULTSOf 66 assessable patients, the overall response rate was 51.5% in the trial group and 36.4% in the control group (P = 0.22). The median TTP was 6.0 months for the trial group, and 3.6 months for the control group (P < 0.001). The score of quality of life in trial group was higher than that in the control group, but no significant difference was observed between the two groups (P > 0.05). There were no significant differences in toxicities between the two groups (P > 0.05).

CONCLUSIONNP regimen combined with thalidomide can significantly prolong the median time to tumor progression in patients with advanced NSCLC. Thalidomide may have a synergic activity with NP regimen without increased toxicities.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; adverse effects ; Disease Progression ; Drug Synergism ; Feeding and Eating Disorders ; chemically induced ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; chemically induced ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Quality of Life ; Remission Induction ; Thalidomide ; administration & dosage ; adverse effects ; Thrombocytopenia ; chemically induced ; Vinblastine ; administration & dosage ; adverse effects ; analogs & derivatives ; Vomiting ; chemically induced

OBJECTIVETo construct different mutants of human p53 for expression in eukaryotic cells and investigate the effects of these mutants on stress-induced cell apoptosis.

METHODSHuman p53 cDNA was amplified by PCR and cloned into pcDNA3/HA vector following the routine procedures. The Ser15 and Ser46 of p53 were mutated to Ala and identified by enzyme digestion and PCR, and these mutants were expressed in NIH3T3 cells and detected by Western blotting. After transfection with the plasmids of different p53 mutants, the NIH3T3 cells were double-stained with AnnexinV-FITC and propidium iodide for apoptotic analysis using flow cytometry.

RESULTSThe recombinant plasmids of HA-tagged wild-type p53, HA-p53(WT), and its mutants, HA-p53(S15A) and HA-p53(S46A), were successfully constructed and expressed efficiently in NIH3T3 cells. The apoptotic ratio of p53(WT)-transfected cells induced by arsenite increased and that of p53(S15A)-transfected cells decreased significantly after arsenite stimulation, but no significant changes occurred in the apoptosis of p53(S46A)-transfected cells.

CONCLUSIONThe phosphorylation on Ser15 of p53 plays an important role in mediating arsenite-induced cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenites ; pharmacology ; Base Sequence ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Mice ; Molecular Sequence Data ; Mutation ; NIH 3T3 Cells ; Phosphorylation ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Objective To compare the diagnostic value 18F-fluorothymidine (FLT) and 18F-fluorodeoxyglucose (FDG) PET/CT in detecting lymph node metastases of untreated thoracic esophageal carcinoma. Methods Twenty-two patients with thoracic esophageal squamous cell carcinoma underwent both 18F-FLT and 18F-FDG PET/CT before surgery. The imaging results of the two modalities in detecting regional lymph node metastases were compared prospectively with the pathologic findings. The X2-test was used with SPS S 13.0. Results All patients underwent esophagectomy and lymphadenectomy. The metastatic lymph nodes were found in 16 patients, from which 47 of 424 excised nodes were positive by pathologic examination. False positive results were 14 while false negative 8 on 18F-FDG PET/CT. In contrast, false positive results were only 3 but false negative were 12 on 18 F-FLT PET/CT. The sensitivity, specificity, accuracy,negative predictive value, and positive predictive value were 74.47% ( 35/47 ), 99.20% ( 374/377 ),96.46% (409/424), 96.89% ( 374/386 ) and 92.11% ( 35/38 ) respectively for 18 F-FLT PET/CT, whereas the corresponding values were 82.98% (39/47), 96.29% (363/377), 94.81% (402/424), 97.84%(363/371 ) and 73.58% (39/53) respectively for 18 F-FDG PET/CT (X2 = 0.572, 6.018, 1.017, 0.348,3.852, P＞0. 05, ＜0.05, ＞0.05, ＞0.05 and ＞0.05). Conclusions Compared with 18F-FDG PET/CT, 18F-FLT PET/CT may be less sensitive but more specific for the detection of lymph node metastases of thoracic esophageal carcinoma.

Objective To investigate the relationship between atherosclerotic lesions of arteries of lower extremities and metabolic disorders in patients with diabetic foot. Methods Three hundreds and sixty two patients with type 2 diabetes were selected, including 232 males and 130 females, with average age of (64.9?11.2) years and the average diabetic duration of (9.2?7.5) years. Atherosclerotic lesions of the arteries were detected by type B ultrasound. According to severity of lesions of femoral, popliteal and tibial arteries, the patients were classified into four groups: A-control group, B-plague formation (plague), C-arterial stenosis (stenosis) (luminal narrowing≥50%) and D-arterial occlusion (occlusion). Fasting blood glucose, GHbAlc and lipid levels (Total cholesteral, TC; Triglyceride, TG; Low density iipoprotein, LDL) were tested in all patients. Results (1)GHbAlc levels in group B and group C were significantly different from that in group A respectively[(8.4?2.2%)vs(7.8?2.2%),P

OBJECTIVETo construct pNTAP-MK2 eukaryotic expression plasmid and establish a HEK293 cell line stably expressing tandem affinity purification (TAP)-tagged MK2.

METHODSThe MK2-encoding region was subcloned into the vector pNTAP to construct the recombinant plasmid pNTAP-MK2, which was subsequently transformed into DH5 alpha.E.coli. After identification by PCR, digestion with restriction endonuclease and sequencing, the recombinant expression plasmid was transfected into HEK293 cells via liposome, and the cell line with stable expression of exogenous TAP tag-MK2 gene was selected by antibiotic G418. The expression and localization of the fusion protein TAP tag-MK2 were detected by Western blotting and immunofluorescence assay.

RESULTSThe results of PCR, restriction endonuclease digestion and sequencing all confirmed the correct construction of the recombinant eukaryotic expression plasmid pNTAP-MK2. Western blotting showed that the recombinant plasmid was expressed stably in HEK293 cells after transfection with G418 selection. Immunofluorescence assay identified the expression product TAP tag-MK2 mainly in the cell nuclei.

CONCLUSIONThe eukaryotic expression vector pNTAP-MK2 has been successfully constructed, and in the established cell line with stable expression of TAP tag-MK2, TAP tag does not influence the localization of exogenous MK2.

Gene Expression ; Genetic Vectors ; HEK293 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Plasmids ; Protein-Serine-Threonine Kinases ; genetics