Objective To construct the expression vector of early growth response-1 promoter-macrophage inflammatory protein 3? (pEgr-1-MIP-3?, pEM) to study MIP-3? expression in transfected Lewis lung cancer cells. Methods MIP-3? cDNA was inserted into plasmid with Egr-1 promoter by directional cloning. The vectors were transfected into Lewis lung cancer cells with liposomes. The expression of MIP-3? in transfected Lewis lung cancer cells induced by electron ray irradiation at different doses were confirmed by RT-PCR and Western blotting. Results The recombinant vector was sequenced, and the result indicated the correct insertion in the expression vector. After electron ray irradiation at different doses, MIP-3? expression in the transfected Lewis lung cancer cells increased significantly. Conclusion Electron ray can enhance the expression of MIP-3? in Lewis lung cancer cells. This has laid the experimental foundation for the studies of pEM for radio-genetic therapy in vivo.

Objective To observe the effects of tissue factor pathway inhibitor(TFPI) on thrombosis formation in rabbit carotid arteries after ballon injury. Methods Fouty rabbits with the weight 2.5-3.0 kg were respectively divided into 4 groups, Ad-TFPI, Ad-LacZ, PBS and normal control groups. The normal control group was not given any treatment and other 3 groups were given 0.2 ml Ad-TFPI, Ad-LacZ or PBS reproduced by the Dispatch catheter respectively after the PTCA balloon iniury on the right carotid arteries. Ten days after gene transfer the repeated balloon injury was performed in the 3 groups, and the first balloon injury was performed in the normal control group by the same method. The carotid blood flow was recovered immediately after the injury. Thirty minutes later all the animals were sacrificed. The injured carotid arteries and one part of contralateral normal artery were cut down, scissored along the long axis, flattened and fixed in the 2% glutaral. The platelet aggregation and thrombosis formation on the luminal surfaces was observed under electron microscope. Results The electron microscope results showed that the vascular endothelial cell structure was integrated and lined up in order in the nomal artery which had no any injury. After the balloon injury in the normal control group, the structure of the endothelial cell was disintegrated, and there was some platelet aggregation but no fibrosis formation. A large amount of platelet aggregated but no fibrosis formed in Ad-TFPI group after the repeated balloon injury. A large amount of fibrosis formed and red cells piled up in the Ad-LacZ and PBS group. The positive rate of thrombosis formation among groups had siginificant differences(χ2=14.95, P<0.01). The positive rate in Ad-TFPI group(20%) was lower than that in Ad-LacZ group(80%, χ2=7.20, P<0.01) and PBS group(70%, χ2=5.05, P<0.05), but was higher than that in the normal control group(10%, χ2=0.39, P>0.05). The positive rate in Ad-LacZ group(80%) was higher than in the normal control group(10%, χ2=9.90, P<0.01) and in the PBS group(70%, χ2=0.27, P> 0.05). The positive rate in PBS group(70%) was higher than that in the normal control group(10%, χ2=7.50, P< 0.01). Conclusions The repeated balloon injury method can cause a large amount of fibrosis formation in the rabbit carotid. TFPI gene inhibits thrombosis formation in balloon-injured rabbit carotid arteries.

Objective To analyze the risk factors for failures of intramedullary and extramedullary internal fixation in surgery of femoral intertrochanteric fractures in elderly patients.Methods A retrospective study was conducted of the 205 elderly patients with osteoporotic femoral intertrochanteric fracture who had accepted closed reduction together with intramedullary and extramedullary internal fixation between September 2005 and August 2014.They were 89 men and 116 women,from 65 to 98 year of age (average,78.8 years).By AO classification,137 cases were of types AI.1-A2.1 (stable fractures),and 68 of type A2.2-A3.3(unstable fractures).The incidence of internal fixation failure and Harris scores at the last follow-up were recorded.The factors possibly contributing to the failure were analyzed using the univariate analysis and multivariate logistic regression analysis.Results Of the patients,192 obtained a mean follow-up of 39 months (from 14 to 60 months),but 13 were lost after a 10-month follow-up.Of the 205 patients,internal fixation failure occurred in 12 (incidence of 5.9％).Five failed cases received DHS fixation and 7 accepted PENA fixation.At the last follow-up when the 13 cases were lost after 10-month follow-up,the mean Harris hip score was 81.6 (from 57 to 92),and the excellent to good rate was 84.9％ (29 excellent cases,145 good ones,10 fair ones and 21 poor ones).The multivariate regression analysis revealed that tip-apex distance (TAD) ＞ 25 mm(OR ＝ 333.33),severe osteoporosis (OR =267.44),AO types A2.2-A3.3 (OR ＝ 22.24),functional reduction of fracture (OR =20.79),and concomitant medical diseases (OR =4.59) were independent risk factors for failures of internal fixation.Conclusions DHS and PFNA fixations are effective treatments for elderly patients with femoral intertrochanteric fractures.TAD＞ 25 mm,severe osteoporosis,unstable factures,functional reduction of fracture,and concomitant medical diseases may lead to internal fixation failure in surgery of femoral intertrochanteric fractures in elderly patients.

Objective To obtain the normal reference ranges of global longitudinal strain (GLS),global circumferential strain (GCS),global area strain (GAS) and global radial strain (GRS) of left ventricular in normal adults by three-dimensional speckle tracking imaging (3D-STI) using Meta analysis.Methods Eligible trials which detected global strain of left ventricular in normal subject through 3D-STI were searched in Embase,Pubmed,Cochrane Library database.According to the heterogeneity,parameters of contained studies were analyzed the weighted mean difference (WMD) and 95％ confidence interval (CI).The statistical software was STATA 12.0.Results Totally 1 552 healthy adults from 27 articles were included.Based on the Meta-analysis,theWMDand 95％CIofGLSwere 17.80 and (16.27,19.33),of GCS were 24.73 and (22.50,26.95),of GRS were 47.86 and (39.52,56.19),of GAS were 36.17 and (34.08,38.26).Conclusion The Meta analysis defines reference range of strains obtained by 3D-STI in healthy adults.Using these parameters of 3D globe strains,a guidance of reference for patienfs management and therapy selection may be provided.

Objective: To evaluate the effects of dietary behaviors on the risk of hypertension, diabetes and cardiovascular diseases. Methods: A total of 12 208 subjects aged 18-60 years old were investigated by questionnaires to collect demographic data, dietary behaviors and lifestyle information, when they did health examination in a tertiary hospital in Beijing from 2014 to 2019. During the observation period of five year, the incidence of hypertension, diabetes and cardiovascular diseases were collected through health examination files every year. The multivariate logistic regression model was employed to analyze the associations of dietary behaviors with hypertension, diabetes and cardiovascular diseases. Results: The study included 6 218 ( 50.93% ) males and 5 990 ( 49.07% ) females. The cumulative incidence rates of hypertension, diabetes and cardiovascular diseases were 7.75%, 2.72% and 3.49%, respectively. The multivariate logistic regression analysis indicated that the high-sodium diet ( OR=1.422, 95%CI: 1.191-1.697 ) , eating fast ( OR=1.457, 95%CI: 1.102-1.974 ), eating more refined grain ( OR=1.251, 95%CI: 1.050-1.490 ) and drinking milk less than once a week ( OR=1.316, 95%CI: 1.022-1.697 ) were risk factors for hypertension. The high-sodium diet ( OR=1.344, 95%CI: 1.048-1.725 ), eating fast ( OR=1.733, 95%CI: 1.046-2.871 ), eating more meat ( OR=1.651,95%CI: 1.263-2.158 ) were risk factors for diabetes. High-sodium diet ( OR=1.501, 95%CI: 1.192-1.889 ) was risk factors for cardiovascular disease. Conclusion The diet with high sodium, more meat and refined grain as well as eating fast can increase the risk of hypertension, diabetes and cardiovascular diseases.

Objective To investigate the in vivo gene expression of adenovirus-mediated human tissue factor pathway inhibitor (hTFPI) and its inhibition effects on intimal proliferation in rabbit carotid arteries after ballon injury. Methods Rabbits underwent carotid artery balloon injuries were treated with Ad-TFPI (n=25), Ad-LacZ (n=25) or PBS (n=10), respectively. Sham operated rabbits (n=10) serve as normal controls. The expressions of human TFPI at mRNA and protein levels were detected by RT-PCR and ELISA respectively on the 3rd, 7th, 10th, 14th, 28th day after operation. Intimal proliferation was detected by angiograms and morphometric analysis. Results TFPI mRNA and protein expressions were detected at 3 days and peaked at the 10th and 14th day after TFPI gene transfer. The expressions were still detectable on the 28th day. There was no TFPI expression in Ad-LacZ group. The carotid angiogram results indicated that the minimal lumen diameter in TFPI group was significantly larger and the lumina stenosis percentage was significantly lower in TFPI group compared those in Ad-LacZ and PBS groups (all P < 0.05). The morphometric analysis showed that the intimal area, the ratio of the intimal/media area, the lumina stenosis percentage in TFPI group were all significantly reduced compared with those in Ad-LacZ and PBS groups (all P<0.01).Conclusions The TFPI gene could be effectively transferred by adenovirus vector to injured carotid arteries in rabbits.

Homoisoflavonoid is a special type in flavonoids. There are more than 110 homoisoflavonoid compounds isolated from natural materials. Homoisoflavonoid compounds show many bioactivities on anti-inflammatory, estrogenicy, antiestrogenic, anticancer and angioprotective etc. This paper summarized the plant sources, structural types spectrocopy features and the biology activities of homoisoflavonoids.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Antimutagenic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Humans ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plants, Medicinal ; chemistry ; Vasodilator Agents ; pharmacology

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

OBJECTIVETo detect the expression of aldehyde dehydrogenase (ALDH) in tongue squamous cell carcinoma Tca8113 cell line and observe the biological characteristics of tumor cells expressing high level of ALDH (ALDH(hig)).

METHODSThe expression of ALDH in tongue squamous cell carcinoma Tca8113 cell line was detected by the flow cytometry. The ALDH(hig) and ALDH(low) subpopulation cells were isolated and cultured in vitro respectively. The ability of differentiation, proliferation and tumorsphere formation were detected. Suppression subtractive hybridization (SSH) was employed to screen the genes overexpressed in ALDH(hig) subpopulation cells.

RESULTSThe ALDH(hig) cells in tongue squamous cell carcinoma Tca8113 cell line was rarely 1.3%. Compared with ALDH(low) cells and unsorted cells, ALDH(hig) cells demonstrated increased proliferation capacity. After culture in vitro, the percentage of ALDH(hig) cells decreased gradually to the level of that in unsorted tumor cells. The ALDH(hig) subpopulation cells could survive in serum free medium and generate tumorspheres. SSH analysis revealed these tumor cells also expressed stem cell related genes.

CONCLUSIONThere is a rare subpopulation cells expressing ALDH highly, which displays characteristics similar to stem cells. ALDH may be one of markers for cancer stem cells of tongue squamous cell carcinoma Tca8113 cell line.

Aldehyde Dehydrogenase ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Neoplastic Stem Cells ; Tongue Neoplasms