OBJECTIVEThis study aimed to investigate the effects of hirudin on the expression of transforming growth factor (TGF-β1) and basic fibroblast growth factor (bFGF) in human gingival fibroblasts (HGFs) in vitro, as well to explore its func- tion in the mechanism of gingival remodeling.

METHODSAfter culturing was performed with classic tissue-explant method, HGFs were derived from normal gingival and gingival hyperplasia tissues followed by orthodontic treatments with different concentrations of hirudin. The mRNA and protein expression levels of TGF-β1 and bFGF were respectively detected by real time quantity polymerase chain reaction and immunocytochemistry.

RESULTSCompared with normal HGFs, TGF-β1 expression promoted collagen synthesis of fibroblasts, whereas bFGF collagen synthesis was decreased in hyperplasia HGFs without hirudin (P < 0.05). Hirudin significantly upregulated the expression levels of bFGF but downregulated TGF-β1 in hyperplasia HGFs (P < 0.05).

CONCLUSIONOrthodontic force may influence the balance of collagen synthesis and degradation in HGFs. Hirudin may modulate the balance of HGF collagen metabolism, thereby promoting gingival remodeling.

Objective To investigate the effects of hepatitis B virus X (HBX) gene on cell morphology and transdifferentiation of human proximal tubular epithelial cells.Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.The expression of HBX was confirmed by Q-PCR and Western blotting.Untransfected HK-2 cells and those transfected with empty vector were used as control.The morphology of HK-2 cells was observed by microscopy,the expressions of differentiation marker proteins α-SMA and E-cadherin were detected by Western blotting and Q-PCR,and the contents of IL-1,IL-6 and TNF-α in the supernatant were detected by ELISA assay.Results HBX was successfully expressed in HK-2 cells after transfection.After transfection of HBX gene,the shape of HK-2 cells became irregular,HK-2 cells significantly expressed E-cadherin and α-SMA,and had high levels of IL-1,IL-6 and TNF-α in the cell supernatant (P＜0.01).Conclusion Overexpression of HBX gene in renal tubular epithelial cells may damage cell morphology and promote the occurrence of epithelial-mesenchymal transdifferentiation,which may be related to the inflammatory microenvironment.

Objective To study the effects of ?-catenin-dependent lymphoid enhancer factor(LEF-1) isoforms on biological behavior of HeLa cells.Methods ?-catenin-dependent LEF-1 genes were obtained by PCR from human lymphoid node cDNA library and inserted into pcDNA3.1/V5-His vector to construct the eukaryotic expression plasmid pcDNA3.1-F-LEF-1.Using lipofectamineTM 2000,the plasmid pcDNA3.1-F-LEF-1 was transfected into Hela cells.Then we screened the stable cell lines that expressed the truncated LEF-1 isoforms by G418 and identified the expression of target gene with Western blot.Then we analyzed the proliferation,apoptosis,cell clone formation and capability of tumor formation in vivo of transfected cell lines.Results We successfully constructed the ?-catenin-dependent LEF-1 eukaryotic expression plasmid and obtained the stable HeLa cell lines that expressed the full-length LEF-1 isoforms.The proliferation and capability of tumor formation in vivo of transfected cells were increased while apoptosis was decreased.Conclusion The overexpression of ?-catenin-dependent isoforms can stimulate the malignant biological behavior of HeLa cells.

Purpose To detect the expression of NKX2. 2 protein in gastrointestinal, pancreatic and esophageal neuroendocrine tumors and the correlation between the expression of NKX2. 2 and the clinicopathologic parameters. Methods 41 cases of gastrointestinal, pancreatic and esophageal neuroendocrine tumors were collected. The expression of NKX2. 2, Syn and CgA in neuroendocrine tumor samples were checked by using immunohistochemical staining. Results The positive expression of NKX2. 2 protein was localized in the nucleus. NKX2. 2 protein showed positive staining in neuroendocrine tumors of the seven original sites. In the four cases of normal pancreatic islet cells also showed strongly diffuse positive nuclear staining. The positive rates of NKX2. 2, Syn and CgA protein in the small intestine, rectum, pancreatic neuroendocrine tumors were 100%, 100% and 46. 7%, respectively. The positive rates of NKX2. 2 in foregut, midgut and hindgut were 30% and 87% (χ2 = 11. 09, P=0. 001). Positive NKX2. 2 protein expression was not associated with gender, age group, grade, tumor size and lymph node metastasis. Conclusion NKX2. 2, as a new type of neuroendo-crine markers, is obviously superior to CgA in the diagnosis of neuroendocrine tumors in the small bowel, rectum and pancreas. NKX2. 2, Syn and CgA, a panel approach may be beneficial to enhance diagnostic accuracy of the neuroendocrine tumors.

Objective To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.Untransfected HK-2 cells and those transfected with empty vector were used as controls.The TLR4 expression was detected by real-time PCR and Western blotting.The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry,and the contents of IL-4 and IFN-γ in the supernatant were detected by EIISA.Results Compared with control groups,the number of apoptotic cells was significantly increased in the HBX transfection group (P ＜ 0.05),and the expressions of TLR4,MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05).IFN-γ level in the supernatant of HBX transfection group was higher (P ＜ 0.05),but IL-4 level was lower as compared to control groups (all P ＜ 0.05).Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and upregulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.

Disaster Planning ; Humans ; Physicians

Objective To investigate the clinical and cytogenetic features of myelodysplastic syndrome (MDS) ,and to investigate the application of the International Prognostic Scoring System Revised (IPSS-R) in myelodysplastic syndrome .Methods A retro-spective analysis was conducted on 112 patients diagnosed with MDS on basis of French-American-British(FAB) and World Health Organization(WHO) criterion .Conventional cytogenetics are performed to investigated the cytogenetics changes .Results Karyo-type abnormalities are found in 46(41 .1% ) patients .Cytogenetics subgroups are devided into 5 groups according to IPSS-R criteri-on :very good ,good ,intermediate ,poor ,very poor ,which account for 2 cases(1 .8% ) ,48 cases (42 .9% ) ,41 cases (36 .6% ) ,10 ca-ses (8 .9% ) ,11 cases (9 .8% ) ,with a median survival time(MS) 59 ,36 ,15 ,10 month respectively ,except for very good .Conclusion It is of great guiding significance of IPSS-R criteria for prognostic stratification .

Objective To investigate the synergistic anticancer effect of ATO combined with Rad001 on human ovarian cancer cells in vitro and the underlying mechanisms. Methods SKOV3 cells were treated with Rad001, ATO and a combination of Rad001 and ATO, respectively. Cell relative luminescence units, viability, and combination index were tested. The apoptosis was quantified by Flow cytometry. Autophagy and apoptosis associated proteins were measured with immunoblotting. Results Significant decrease in live cell number were observed in cells treated with combination of Rad001 and ATO , compared with single compound treatment (P <0.05). Moreover, a higher rates of autophagy as well as apoptosis were observed in the combination treatment compared with single compound treatment (P < 0.05). Conclusions combination of Rad001 and ATO can result in synergistic cytotoxicity through activation of the excessive autophagy and apoptotic pathway.

Objective To explore the effect of scoliosis-specific exercises ( SSE) on patients with mild ado-lescent idiopathic scoliosis ( AIS) . Methods Thirty patients with mild AIS were assigned to a control group ( n=10) or an SSE group ( n=20) . The control group received routine health education, while the SSE group completed a 60-minute set of SSE 2 to 3 times a week for 12 weeks. The angle of trunk rotation ( ATR) , maximum Cobb angle and angle of vertebral rotation (AVR) were recorded. Bone strength parameters including the speed of sound (SOS), Z-score and percentile distal radius were measured. Surface electromyography ( sEMG) was performed for the erector spinae muscle. In addition, forced vital capacity ( FVC) and forced expiratory volume in one second ( FEV1) were measured and compared with the predicted values ( FVC/pred% and FEV1pred%) . A falling index ( FI) and quality of life ( QOL) were measured. Results Compared with before the treatment, the average maximum Cobb angle in the control group increased significantly after the lessons, but there was no significant difference in any of the other measures, including QOL. For the control group the activation rate of the concave side of the apex level erector spi-nae was significantly lower than on the convex side both before and after the lessons. The SSE group showed no sig-nificant improvement in their average ATR, maximum Cobb angle, AVR or FI results, but their average SOS, Z-score and percentile of the distal radius, FVC, FEV1 and motor function improved significantly after the treat-ment. Before the treatment the activation rate of their concave side was also lower than on the convex side, but after the treatment there was no significant difference between them on average. Conclusion Early SSE can prevent further deformation, promote bone strength, improve lung function reduce the difference in the motor control of the bilateral erector spinae muscles among patients with mild AIS. It can promote a better quality of life and is worth ap-plying in clinical practice.