Intra-plaque angiogenesis plays an important role in the development of atherosclerotic plaque. Vascular endothelial growth factor (VEGF) is a major initiating factor in this pathologic progress. One selective and specific inhibitor of VEGF is soluble VEGF receptor-1 (sFlt-1). The anti-angiogenic utilization of sFlt-1 in treatment of atherosclerotic plaque has not been fully confirmed yet. Our study was designed to construct eukaryotic expression recombinant pEGFP-N1-sFlt-1, evaluate sFlt-1 recombinant's effects on endothelial cells proliferation and tube formation in vitro, and investigate effects of local high-expressed sFlt-1 on atherosclerotic plaque in vivo. Rabbit models of atherosclerotic plaque were established by high-lipid diet combined with injury induced by balloon catheter on iliac artery intima. Animals were divided into four groups randomly: control group (C), atherosclerotic plaque group (AP), atherosclerotic plaque with blank vector pEGFP-N1 transfection group (APV), and atherosclerotic plaque with pEGFP-N1-sFlt-1 transfection group (APsFlt-1). The local expression of sFlt-1 protein in target artery was detected by western blotting. The plaque area (PA), plaque circumference (PC), and maximum plaque thickness (MPT) were measured via HE staining. Degree of intra-plaque angiogenesis was evaluated by CD34+ cells immunohistochemistry. As results, we observed that pEGFP-N1-sFlt-1 transfection suppressed the HUVECs proliferation and ability of tube formation, against the effect of VEGF. We obtained higher local expression of sFlt-1 protein in Group APsFlt-1 than that in other groups (P < 0.05). PA, PC, and MPT of plaque in group APsFlt-1 were significantly decreased when compared with other groups (P < 0.05). Amount of annulations surrounded by CD34-positive cells was significantly decreased in pEGFP-N1-sFlt-1 transfection group, which represented decreased level of intra-plaque neovessels formation. The present study confirmed that local gene delivery of sFlt-1 can suppress plaque formation, as one of possible mechanisms, via inhibitive effect on intra atherosclerotic plaque angiogenesis, which hints at the clinical utility of sFlt-1 in atherosclerosis therapy.

Objective To treat superficial hemangioma in children according to the local blood flow.Methods A total of 98 children with superficial hemangiomas admitted to our hospital from January 2005 to June 2009, and their clinical data were analyzed.Results According to the local blood flow velocity, 98 children were treated with injections or injection plus surgical treatment, respectively. Ninety-four children (95.9%) were cured. Conclusion Injection therapy is effective for children with superficial hemangioma, but we should arrange individualized treatment according to the local blood flow in children.

The tumour suppressor p53 modulates transcription of a number of target genes involved in cell cycle arrest, apoptosis, DNA repair, and other important cellular responses. Mitofusin-2 (Mfn2) is a novel suppressor of cell proliferation that may also exert apoptotic effects via the mitochondrial apoptotic pathway. Through bioinformatics analysis, we identified a p53 binding site in the Mfn2 promoter. Consistent with this, we showed that the p53 protein binds the Mfn2 promoter directly both in vitro and in vivo. Additionally, we found that Mfn2 mRNA and protein levels are up-regulated in a p53-dependent manner. Furthermore, luciferase assays revealed that the activity of the wild-type Mfn2 promoter, but not a mutated version of the promoter, was up-regulated by p53. These results indicate that Mfn2 is a novel p53-inducible target gene, which provides insight into the regulation of Mfn2 and its associated activities in the inhibition of cell proliferation, promotion of apoptosis, and modulation of tumour suppression.

Mitochondrial GTPase mitofusin-2 gene (Mfn2) is a novel gene characterised as a cell proliferation inhibitor. Mfn2 protein over-expression, mediated by an adenovirus, has a significant anti-tumour effect in A548 and HT-29 cells. However, there is no report on the effect of Mfn2 on urinary bladder carcinoma (UBCC). In this study, we sought to investigate the function of Mfn2 in UBCC. Mfn2 expression in 36 paired UBCC samples was investigated by reverse transcription-polymerase chain reaction and Western blot analyses. An adenovirus encoding the complete Mfn2 open reading frame (Ad-Mfn2) was used to infect UBCC cells, and an adenoviral vector encoding green fluorescent protein (Ad-GFP) was used as a control. The effects of Mfn2 on cell-cycle distribution and apoptosis were assessed by flow cytometry and Western blot analyses. The Mfn2 protein showed significantly lower expression in UBCC tissues than nearby non-tumourous tissues. Ad-Mfn2 exhibited a significant anti-tumour effect in T24 and 5,637 cells. Mfn2 overexpression in T24 cells significantly inhibited cell proliferation, by arresting the transition of the cell cycle from the G(1) to S phase, and induced apoptosis by upregulating active caspase-3 and cleaved PARP levels. Mfn2 also induced increased p21 and p27 expression levels, but down-regulated PCNA levels. These findings indicate that Mfn2 is a potential UBCC tumour suppressor gene, which showed significantly lower expression in tumour tissues than adjacent non-tumourous tissues and could promote apoptosis and inhibit the proliferation of UBCC cells. Mfn2 may become an important therapeutic target for treating UBCC.

Membrane fusion and fission in intracellular trafficking is controlled by both intraluminal Ca(2+) release and phosphoinositide signaling. However, the molecular identities of the Ca(2+) release channels and the target proteins of phosphoinositides are elusive. Here, by direct patch-clamping of the endolysosomal membrane, we report that PI(3,5)P(2), an endolysosome-specific phosphoinositide, binds and activates endolysosome-localized mucolipin TRP (TRPML) channels with specificity and potency. Both PI(3,5)P(2)-deficient cells and cells that lack TRPML1 exhibited enlarged endolysosomes/vacuoles and trafficking defects in the late endocytic pathway. We find that the enlarged vacuole phenotype observed in PI(3,5)P(2)-deficient mouse fibroblasts is suppressed by overexpression of TRPML1. Notably, this PI(3,5)P(2)-dependent regulation of TRPML1 is evolutionarily conserved. In budding yeast, hyperosmotic stress induces Ca(2+) release from the vacuole. Here, we show that this release requires both PI(3,5)P(2) production and a yeast functional TRPML homolog. We propose that TRPMLs regulate membrane trafficking by transducing information about PI(3,5)P(2) levels into changes in juxtaorganellar Ca(2+), thereby triggering membrane fusion/fission events.

Acute viral myocarditis (AVMC) is characterized by virus-triggered myocardial inflammation, and Coxsackievirus B3 (CVB3) is the primary pathogen. We previously proved that Th17 cells, besides having proinflammatory effects, were involved in AVMC by enhancing humoral response. However, the relationship between Th17 cells and CVB3 replication remains unknown. In this experiment, we infected BALB/c mice with CVB3 for establishing AVMC models and then found that, with the increase of viral replication, the expressions of splenic Th17 cells, serum IL-17, and cardiac IL-17 mRNA were elevated significantly, accompanied by the progressive cardiac injuries of AVMC. Furthermore, on day 5, the peak time for viral replication, correlation was positive between cardiac IL-17 mRNA and CVB3 RNA (correlation index = 0.835; p < 0.01). Although the expressions of Th1 and CD8(+) T cells, which could secrete the antiviral cytokine IFN-gamma and damage the heart, were also elevated, along with Th17 cells, in AVMC, the neutralization of IL-17 further upregulated the percentages of splenic Th1 and CD8(+) T cells and the levels of cardiac IFN-gamma mRNA. The cardiac pathological changes were obviously improved after neutralization, with reduced viral replication followed by decreases in the cardiac inflammatory cytokines IL-17, TNF-alpha, and IL-1beta. These data suggest that Th17 cells contribute to CVB3 replication in AVMC, and that IL-17 might be an important target for regulating the balance of antiviral immunities.

One hundred and sixty-three Bordetella bronchiseptica isolates from pigs, consisting of 65 isolated in 2003 and 98 isolated in 2007, were tested for antimicrobial susceptibility to 17 antimicrobial agents by the disk diffusion method. All 163 B. bronchiseptica isolates were sensitive to polymyxin B and cefoperazone/sulbactam; the majority of the strains were sensitive to amikacin (149/163; 91.4%), gentamicin (132/163; 81.0%), ampicillin/sulbactam (127/163; 77.9%) and ciprofloxacin (115/163; 70.6%). A high level of resistance was found for furazolidone (100%), ampicillin (90.2%), cefazolin (89.0%), streptomycin (87.7%), amoxicillin/clavulanic acid (74.2%), and tetracycline (64.4%). Comparison of the data revealed that isolates with multi-resistance to at least six or eight of 17 antimicrobials used became more frequent from 32.3 or 16.9% in 2003 to 90.8 or 41.8% in 2007.

Mps1 is a dual-specificity protein kinase with key roles in regulating the spindle assembly checkpoint and chromosome-microtubule attachments. Consistent with these mitotic functions, Mps1 protein levels fluctuate during the cell cycle, peaking at early mitosis and abruptly declining during mitotic exit and progression into G1 phase. While evidence in budding yeast indicates that Mps1 is targeted for degradation at anaphase by the APC/cCdc20 complex, little is known about the regulatory mechanisms that govern Mps1 protein levels in human cells. Here, we provide evidence for the ubiquitin ligase/proteosome pathway in regulating human Mps1 levels during late mitosis through G1 phase. First, we showed that treatment of HEK 293T cells with proteosome inhibitor MG132 resulted in an increase in both the polyubiquitination and the accumulation of Mps1 protein levels. Next, Mps1 was shown to co-precipitate with APC and its activators Cdc20 and Cdh1 in a cell-cycle dependent manner. Consistent with this, overexpression of Cdc20 or Cdh1 led to a marked reduction of endogenous Mps1 levels during anaphase or G1 phase, respectively. In contrast, depletion of Cdc20 or Cdh1 by RNAi treatment both led to the stabilization of Mps1 protein during mitosis or G1 phase, respectively. Finally, we identified a single D-box motif in human Mps1 that is required for its ubiquitination and degradation. Failure to appropriately degrade Mps1 is sufficient to trigger centrosome amplification and mitotic abnormalities in human cells. Thus, our results suggest that the sequential actions of the APC/cCdc20 and APC/cCdh1 ubiquitin ligases regulate the clearance of Mps1 levels and are critical for Mps1 functions during the cell cycle in human cells.

In this article, the dielectrophoretic (DEP) assembly of chemically-modified silica nanoparticles (SiNPs) was introduced. Five types of surface-modified SiNPs, including OH-SiNPs, COOH-SiNPs, CH(3)HPO(2)-SiNPs, PEG-SiNPs, and NH(2)-SiNPs, have been investigated. After applying an ac field with relatively high intensity and frequency, it was shown that only COOH-SiNPs and CH(3)HPO(2)-SiNPs could be self-assembled on the microelectrodes by the DEP forces. The results indicated that the anionic group modification could obviously enhance the DEP self-assembly of SiNPs on the microelectrodes. Then the DEP assembly of CH(3)HPO(2)-SiNPs was selected as a representative to be investigated further. By using Rubpy dye doped in the core of the CH(3)HPO(2)-SiNPs, the assembly process was visualized in real time by inverse fluorescence microscopy. Precise control over the frequency of the applied ac field showed that the DEP forces can assemble CH(3)HPO(2)-SiNPs from aqueous suspensions into submicrowires, and it was found that the number of assembled submicrowires between the microelectrode gaps could be well controlled with reversibility. Furthermore, the DEP assembly process of CH(3)HPO(2)-SiNPs was sensitive to the pH of the dispersed medium. These findings would provide a way to circumvent the difficulty in controlling the dielectrophoretic assembly process of nanoparticles and offer application opportunities for the DEP assembly of chemically modified SiNPs.

OBJECTIVES: Femoral head osteonecrosis (ON) is a serious complication of steroid administration. We examined whether implantation of autologous bone marrow derived endothelial progenitor cells (EPCs) can augment neovascularization and bone regeneration in steroid-induced osteonecrosis of the femoral head.METHODS: Forty 12-week-old male New-Zealand white rabbits were divided into Group I (left untreated, n=12), Group II (core decompression, n = 12) and Group III (core decompression + autologous EPCs implantation, n = 16) after receiving an established inductive protocol for inducing steroid-associated ON. Four weeks later, these rabbits were euthanized, bilateral femora were dissected for Micro-CT-based micro-angiography to assess vascularization, and then the osteonecrotic changes and repair processes were examined histopathologically.RESULTS: Quantitative analysis showed that new vessel formation in Group III was significantly greater compared with other groups at 4 weeks after treatment. The histologic and histomorphometric analysis revealed that the new bone volume was significantly higher in the group III than in the group I and II 4 weeks after treatment.CONCLUSION: A combination of EPCs and core decompression enhances the neovascularization and bone regeneration in rabbit steroid induced femoral head osteonecrosis. Local implantation of endothelial progenitor cells (EPCs) may provide a novel and effective therapeutic option for early corticosteroid-induced ON.

A facile method has been developed to encapsulate Fe(3)O(4) nanocrystals (NCs) in morphology-tunable fibers (belt-like, solid, and tubal) by using a sonochemistry driven synthesis and a subsequent reflux procedure. By adapting the use of tetraethyl orthosilicate, ammonia, Cd(2+), and thiolglycolic acid (TGA) to an ultrasound-driven synthesis, the Fe(3)O(4) NCs were coated with a thin composite shell. Supersonic treatment plays an important role to prevent the agglomeration of the Fe(3)O(4) NCs in an alkaline condition. The composite shell became thicker due to the deposition of SiO(2) monomers, Cd-TGA clusters, Cd(2+), and free TGA molecules during reflux. In addition, these composite shell-coated Fe(3)O(4) NCs were assembled in composite fibers which were created by the growth of Cd-TGA clusters and the deposition of SiO(2) monomers. The Fe(3)O(4) NCs mono-dispersed in fibers revealed superparamagnetic behavior. The magnetic saturation value of tubal fibers is lower than those of belt-like and solid fibers. These fibers with Fe(3)O(4) NCs would be utilizable for further application. The strategy described here should give a useful enlightenment for the design and fabrication of morphology-tunable fibers with functional NCs.

BACKGROUND: Video-assisted mini-thoracoscopy (VAMT) has been used for pulmonary lobectomy for 20 years, which has many merits including small wound, less pain and quick recovery. VAMT is the tendency of pectoral minimally invasive surgical treatment. The aim of this study is to evaluate the efficacy of video-assisted mini-thoracoscopy (VAMT) for radical resection of pleomorphic carcinoma of the lung. METHODS: Complete resection of pleomorphic carcinoma of the lung was performed on 28 patients by VMAT. A 1.5 cm and a 6 cm to 8 cm incision was made during the operation. Both standard surgical instruments and thoracoscopic set were used to treat the pulmonary vessels, perform lobotomy, and remove the lymph nodes in the mediastinum and pulmonary portal. RESULTS: The operation was completed in all of the cases. No peri-operative death occurred. The total volume of hemorrhage was 200 mL to 450 mL (mean, 300 mL). The patients received chest drainage for 3 to 8 days after the operation (mean, 5 d). The time in hospital was 7 d-14 d (mean, 12 d). Five-year overall survival and disease-free survival were 39.2% and 47.1%, respectively. Follow-up was available in all 28 patients for up to 2 to 91 months. Among the 28 cases, only 2 patients died 2 months after the operation, the rest all live up over 1 year. 3-year survival rate was 60.7% (95%CI: 40.3%-81.1%). CONCLUSIONS: VAMT is effective for radical resection of pleomorphic carcinoma of the lung in a short term. Combination the superiority of traditional procedure and VATS, so VAMT is safe and reliable for radical resection of pleomorphic carcinoma of the lung.

There are approximately 33 million injuries involving musculoskeletal tissues (including tendons and ligaments) every year in the United States. In certain cases the tendons and ligaments are damaged irreversibly and require replacements that possess the natural functional properties of these tissues. As a biomaterial, collagen has been a key ingredient in tissue engineering scaffolds. The application range of collagen in tissue engineering would be greatly broadened if the assembly process could be better controlled to facilitate the synthesis of dense, oriented tissue-like constructs. An electrochemical method has recently been developed in our laboratory to form highly oriented and densely packed collagen bundles with mechanical strength approaching that of tendons. However, there is limited information whether this electrochemically aligned collagen bundle (ELAC) presents advantages over randomly oriented bundles in terms of cell response. Therefore, the current study aimed to assess the biocompatibility of the collagen bundles in vitro, and compare tendon-derived fibroblasts (TDFs) and bone marrow stromal cells (MSCs) in terms of their ability to populate and migrate on the single and braided ELAC bundles. The results indicated that the ELAC was not cytotoxic; both cell types were able to populate and migrate on the ELAC bundles more efficiently than that observed for random collagen bundles. The braided ELAC constructs were efficiently populated by both TDFs and MSCs in vitro. Therefore, both TDFs and MSCs can be used with the ELAC bundles for tissue engineering purposes. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.

Objective To survey on men who have sex with men (MSM) population's sexual behaviors, condom-service condition, HIV related knowledge and other issues among MSM population at sexually transmitted disease (STD) clinics to understand the characteristics of behaviors and offer suggestions for effective health education and behavior intervention.Methods From January to December, 2009, we used anonymous questionnairres which involved in their mastery of demographic characteristics, sexual behavior, condom-service condition, HIV related knowledge, and so one, to investigate 200 MSM at 3 STD clinics of comprehensive hospital. Results The average age of informant was (26.7+/- 8.9) years and 121 individules (62.6%) had confirmed with STD in recent one year. In the recent 6 months, the average number of homosexual partners was 9.2+/- 4.8 and 102 (52.8%) had heterosexual partners. In the sexual intercourse with homosexual, 123 individules had anal intercourse (63.7 %) and 117 had oral intercourse (60.6%). In the sexual intercourse with heterosexual, 92 (90.2%) individules had vaginal intercourse, 37 (36.2%) had anal intercourse, and 59 (57.8%) had oral sex behavior.There were a statistical difference between heterosexual and homosexual sex behaviors (P<0.01). The condom-using frequency had statistic difference in different sexual behaviors(chi(2)=188.396, P<0.001). There was no linear correlation between HIV related knowledge and condom-using condition in sexual behaviors except the heterosexual anal intercourse. Conclusion High AIDS knowledge mastery rate is found in our survey. The respondents get HIV/AIDS knowledge through various ways actively. There is no obvious relation between the mastery of HIV related knowledge and condom-using frequency. The ratio of non-protected sexual behaviors is high in heterosexuals. How to adopt effective methods for behavior intervention to MSM at STD clinic needs to be further studied.

Phosphoribosylaminoimidazole-succinocarboxamide synthase (SAICAR synthase) plays an essential role in the de novo biosynthesis of purine nucleotides. In this study, the SAICAR synthase from Streptococcus suis was cloned and overexpressed in Escherichia coli. The subsequent product was purified and crystallized using the hanging-drop vapour-diffusion method. The crystals diffracted to 2.8 A resolution and belonged to space group P2, with unit-cell parameters a = 70.2, b = 52.2, c = 153.9 A, beta = 102.8 degrees .

A simple method for the preparation of monodisperse protein-loaded polymer microspheres is presented in this paper. The method is based on the co-extrusion of an internal phase of an aqueous protein solution and an external phase of an organic polymer solution through a 200-micron-sized hole. Controlled in the correct flow region, this process produces a core-shell-structured laminar liquid jet, which breaks to form monodisperse compound liquid droplets. Stabilized in a dilute aqueous polyvinyl alcohol (PVA) solution, the droplets are converted into solid protein-loaded polymer microspheres through evaporation of the organic solvent. Results show that preparation parameters such as polymer concentration, total flow rate, flow rate ratio of the aqueous to organic phase have significant effects on the mean particle size, particle morphology and protein encapsulation efficiency (EE). The results of biodegradation and the protein release characteristics of the polymer microspheres are also presented.

Cyclins are primary regulators of the activity of cyclin-dependent kinases and play crucial roles in cell cycle progression in eukaryotes. Although extensive studies have revealed the roles of some cyclins and underlying mechanisms in plants, relatively few cyclins have been functionally analyzed in maize. We identified 59 cyclins in the maize genome, distributed on 10 chromosomes; these were grouped into six types by phylogenetic analysis. The cyclin genes in the maize genome went through numerous tandem gene duplications on five chromosomes. However, no segmental duplications, which occur in rice, were found on maize chromosomes. This information allows us to assess the position of plant cyclin genes in terms of evolution and classification, which will be useful for functional studies of maize cyclins.

An integrated single-tooth modeling scheme is proposed for the 3D dental model acquired by optical digitizers. The cores of the modeling scheme are fusion regions extraction, single tooth shape restoration, and single tooth separation. According to the "valley" shape-like characters of the fusion regions between two adjoining teeth, the regions of the 3D dental model are analyzed and classified based on the minimum curvatures of the surface. The single tooth shape is restored according to the bioinformation along the hole boundary, which is generated after the fusion region being removed. By using the extracted boundary from the blending regions between the teeth and soft tissues as reference, the teeth can be separated from the 3D dental model one by one correctly. Experimental results show that the proposed method can achieve satisfying modeling results with high-degree approximation of the real tooth and meet the requirements of clinical oral medicine.

To investigate the association of expression and promoter methylation of tumor-suppressor genes with risk of ovarian cancer, we conducted a case-control study of 102 patients with serous epithelial ovarian cancer and 100 patients without ovarian cancers. We measured mRNA expression levels (by real-time reverse transcription polymerase chain reaction) and methylation status (by methylation-specific polymerase chain reaction) of five candidate genes (BRCA1, BRCA2, hMLH1, MGMT, and DNMT3B) in tumors from the cases and normal ovaries from the controls. We found that mRNA expression levels of the five genes were decreased in tumors than in normal ovaries with 0.39-fold for BRCA1, 0.25-fold for BRCA2, 0.42-fold for hMLH1, 0.45-fold for MGMT, and 0.87-fold for DNMT3B, calculated by the 2(-DeltaDeltaCT) method. Ovarian cancer risk (odds ratios, ORs) was associated with low expression of all genes (2.95 [95% confidence interval (CI), 1.51 - 5.78] for BRCA1, 3.65 (95% CI, 1.82 - 7.30) for BRCA2, 5.25 (95% CI, 2.52 - 10.96) for hMLH1, and 4.72 (95% CI, 2.32 - 9.62) for MGMT) but not DNMT3B. However, methylation status was not associated with gene expression levels in the tumors, except for hMLH1 whose mean (+/- SD) gene expression was significantly lower in methylated (13.0 +/- 7.6) than in unmethylated (31.2 +/- 44.8) tumors (P < 0.001). We concluded that low mRNA expression of these tumor-suppressor genes, likely due to molecular mechanisms in addition to the promoter methylation in some instances, may be a biomarker for ovarian cancer risk in this study population. Larger studies are needed to validate our findings.

RNA interference (RNAi) libraries have been proven to be a powerful tool for large-scale functional genetic screens. To facilitate high-throughput functional genetic screens in embryonic stem cells, a system for construction of random dsRNA expressing RNAi libraries was developed. Previous studies have demonstrated that sequence-specific gene silencing could be induced by long dsRNA in mouse embryos, mouse oocytes, embryonic stem cells, and some other mammalian cells. Our study demonstrated that the dsRNA interference library can be used for functional genetic screens of genes involved in self-renewal of embryonic stem cells (ES cells). The random RNAi library is easy to construct and provides a useful tool for investigation of molecular mechanisms of cellular development and differentiation.