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 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.

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.

Disaster Planning ; Humans ; Physicians

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.

Objective To investigate the effect of astragalus polysaccharide (AP) on proliferation and apoptosis of human acute myelocytic leukemia cell line KG-1a and the underlying mechanisms.Methods Human acute myelocytic leukemia cell line KG-la was cultured under 37 ℃ and 5 ％ C02,when appropriate cell passage and cryopreservation were performed.After treatment for 48 h with different concentrations of AP,the proliferative activity and apoptosis rate of KG-1a cells were examined by CCK-8 assay and flow cytometry,respectively.The expressions of p-Akt and bcl-2 protein in AP-treated KG-1a cells were evaluated by Western blot.The expression of PTEN mRNA by quantified real-time PCR.Results The proliferative activity of KG-la cell was obviously suppressed by AP treatment,and the inhibition rate increased in a dosedependent manner.The flow cytometry showed that,compared with the control group,the apoptosis rates of KG-1a cells were significantly increased after treatment with AP for 48 h.The early apoptosis rates were (1.98±0.16) ％,(12.60±0.48) ％,(16.31±0.73) ％,the late apoptotic rates were (3.11±0.19) ％,(17.17±1.40) ％,(21.17 ± 0.81)％,the differences were statistically significant (P ＜ 0.05).Western blot showed that the expressions of p-Akt and bcl-2 protein in KG-1a cells decreased significantly after treatment with AP (P ＜ 0.05).In contrast,the mRNA level of PTEN increased (P ＜0.05),which was shown by real-time PCR.Conclusion AP could repress cellular proliferation activity of KG-1a cells,which could be attributed to inhibition of PTEN-PI3K-Akt signaling pathway.

Since the establishment of the People’s Republic of China, the development of China’s official de-velopment assistance for health has been going through three phases. To date, it has developed in many forms, inclu-ding the dispatch of medical team and the construction of health facilities. Since the market reforms, the global con-text together with the domestic socioeconomic foundations have changed; the impact of China’s relatively simple and segmented development assistance for health on the development of health systems in developing countries is limited;the effectiveness of assistance has been watered-down due to segmentation and vague management and accountability systems, as well as the lack of an overarching strategy;China’s health institutions, techniques and products suffer va-rious obstacles in their transfer to other countries where rules are dominated by western countries;compared with the increasing and multiple demands of development assistance for health from developing countries, the capacities of co-operation need to be further developed. As the paper suggests, use the “new major-country” and “new morality-in-terest” and“human oriented” concepts, as well as the ideology of“aid in order to develop, develop in order to coop-erate, so as to develop hand-in-hand” to guide China’s development assistance for health;innovate stereo-aid models to adapt to the changed foreign and domestic socioeconomic context; reform the development assistance for health management system and define rights and obligations appropriately;strengthen coordination and information sharing;link development assistance for health with global health governance to promote a maximized spillover effect;mobilize the civil society with strengthened guidance and supervision.