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.

Fibroblast Growth Factor 2 ; Fibroblasts ; Gingiva ; Hirudins ; Humans ; RNA, Messenger ; Transforming Growth Factor beta ; Transforming Growth Factor beta1

Disaster Planning ; Humans ; Physicians

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 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.

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 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.

OBJECTIVETo investigate the influence of isoliensinine (IL) on CYP3A enzyme activity.

METHODSA mixture metabolic system of liver microsome enzymes in vitro was developed. A HPLC method to test the metabolic activity of CYP3A was established with testosterone as a probe. The activities of CYP3A enzymes were measured with different IL concentrations and incubation time with testosterone.

RESULTSIn mixture metabolic system of liver microsome enzymes, the best incubation concentration of testosterone was 200 μmol/L, the best incubation time was 210 min, in this condition the IC₅₀ of IL for CYP3A inhibition was >1 000 μmol/L.

CONCLUSIONNo significant interaction between IL and CYP3A is detected, which indicates that IL might be used with CYP 3A enzyme substrates.

Animals ; Cytochrome P-450 CYP3A ; metabolism ; In Vitro Techniques ; Isoquinolines ; pharmacology ; Male ; Microsomes, Liver ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; pharmacology

Objective To investigate the clinical outcome of correction of unilateral delayed orbitozygomatic complex fractures with digital guide plates.Methods The study involved 14 cases of unilateral delayed orbitozygomatic complex fractures treated between September 2007 and March 2012.Craniofacial structures were measured by thin-section CT before operation,followed by data input to the image processing software.Before and after the processing of images,three dimensional skull models were produced using rapid prototyping technique and applied to have surgical simulation.Digital guild plates were produced for guiding the reduction of fractures and orbital walls were reconstructed as well.Results All cases were followed up for a period of 6-11 months,which showed basic restoration of facial structure and significant correction of diplopia and enophthalmos,with no infection or dislocation of implants buried in orbit floor.Mild ectropion of the lower eyelid occurred in one case and temporary decrease of frontal wrinkle in two cases,but all recovered in 3-6 months postoperatively.Conclusion Digital guide plate is easy in operation and reliable for treatment of unilateral delayed orbitozygomatic complex fractures.

Objective The palliation of dysphagia in metastatic esophageal cancer remains a challenge ,and the optimal approach for this difficult clinical scenario is not clear .We therefore sought to define and determine the efficacy of various treatment options used at our institution for this condition .Methods Methods We reviewed a prospective database for all patients managed in an e-sophageal cancer referral centre over a 5-year period .All patients receiving palliation of malignant dysphagia were reviewed for de-mographics ,palliative treatment modalities ,complications ,and dysphagia scores (0= none to 4= complete) .The Wilcoxon signed rank test was used to determine significance (P<0 .05) .Results During 2005~2010 ,80 patients with inoperable esophageal cancer were treated for palliation of dysphagia .The primary treatment was radiotherapy in 66% ,metal stenting in 21% and radiotherapy combined with stent in 13% .Mean duration of treatment was 1 day in he stent group and 40 days in the radiotherapy group(P=0 . 001) .In patients treated initially by stenting ,dysphagia improved within 2 weeks of treatment in 82% of patients(dysphagia score of 0 or 1) .However ,18% of patients presented with recurrence of dysphagia at 10 weeks of treatment .In the radiotherapy group , the onset of palliation was slower ,with only 50% of patients palliated at 2 weeks(dysphagia score of 0 or 1) .However ,long-term palliation was more satisfactory ,with 90% of patients remaining palliated after 10 weeks of treatment .Conclusion In inoperable e-sophageal cancer at our centre ,radiation treatment provided durable long-term relief ,but came at a high price of a long wait time for initiation of treatment and a long lag time between initiation of treatment and relief of symptoms .On the other hand ,stenting pro-vided more rapid and effective early relief from symptoms ,but was affected by recurrence of dysphagia in the long-term .