Objective To explore an algorithm to define the threshold value for tumor contouring on 18F-fluorodexyglucose (FDG) PET imaging. Methods A National Electrical Manufacturing Association (NEMA)NU 2 1994 PET phantom with 5 spheres of different diameters were filled with 18F-FDG. Seven different sphere-to-background ratios were obtained and the phantom was scanned by Discovery LS 4. For each sphere-to-background ratio, the maximum standardized uptake value ( SUVmax ) of each sphere, the SUV of the border of each sphere ( SUVborder ), the mean SUV of a 1 cm region of background (SUVbg) and the diameter (D) of each sphere were measured. SPSS 13.0 software was used for curve fitting and regression analysis to obtain the threshold algorithm. The calculated thresholds were applied to delineate 29 pathologically confirmed lung cancer lesions on PET images and the obtained volumes were compared with the volumes contoured on CT images in lung window. Results The algorithm for defining contour threshold is TH% = 33.1% + 46.8% SUVbg/SUVmax + 13.9%/D ( r = 0.994) by phantom studies. For 29 lung cancer lesions, the average gross tumor volumes ( GTV ) delineated on PET and CT are ( 7.36 ± 1.62 ) ml and (8.31 ±2.05) ml, respectively (t = -1.26, P＞0.05). Conclusion The proposed threshold algorithm for tumor contouring on PET image could provide comparable GTV with CT.

AlM:To research the reasonability of current criteria on distant vision anddiopter for recruiting civil aviators in China.
METHODS: The data about distant vision and diopter of 1901 aviators, including flight majors and aviators in active service participating in physical examination of Civil Aviation Flight University of China from 2006 to 2013, were collected. ANOVA and LSD were used to compare the differences between distant vision and diopter among different groups. The Spearman correlation coefficients of distant vision (≥0. 1 vs ≥0. 3) and diopter (0. 00 to -3. 00D) were calculated.
RESULTS: The diopter of civil aviators in China increased with distant vision decreased. The correlation between distant vision and diopter ( 0. 00 to -3. 00D ) among distant vision≥0. 3's population (0. 4 CONCLUSlON: The current eye standard of recruiting civil aviators in China is reasonable to consider both distant vision and diopter, but the standard for distant vision is higher. Further research is proposed to explore the proper standard of distant vision that matches the diopter standard.

Amyloidosis is less involved in the respiratory tract. The tracheobronchial amyloidosis is rare, and the amyloid changes involved in the larynx, trachea and bronchus are more rare. The study introduces a case of the primary laryngeal and tracheobronchial amyloidosis. The patient was diagnosed as primary laryngeal amyloidosis, and had tracheobronchial amyloidosis after 15 years. Due to the rare and non-specific clinical manifestations of the disease, the patient was misdiagnosed as upper respiratory tract infection and bronchial asthma.After the treatment of the clamp of the bronchoscope and argon knife,the symptoms were significantly relieved in the patient.At present,the disease is stable and has not relapsed,but it still needs to track the long-term prognosis.

AIMTo demonstrate the specific killing of folate receptor (FR)-positive tumor cells can be achieved by folate-targeted penicillin-G amidase (PGA) combined with its prodrug substrate N-(phenylacetyl) doxorubicin (DOXP).

METHODSFolic acid was covalently linked to PGA and folate content value was determined by quantitative UV spectrophotometry. The ability of folate conjugated PGA to hydrolyze DOXP was measured by RP-HPLC. Visual demonstration of uptake by FR (+) HeLa and SKOV3 cells was detected by using FITC labeled folate-PGA and a fluorescence microscopy. The cytotoxicity of DOXP towards the cells in the presence or absence of folate-PGA was assayed by using MTT method.

RESULTSThe folate-PGA has a specific activity of 29. 8 U x mg(-1) (protein). FR selectivity was confirmed by fluorescence microscopy. The combination of DOXP prodrug with folate-PGA generated higher cytotoxicity towards the FR (+) cells than free doxorubicin. The IC50 was 0.72 micromol x L(-1) for HeLa cells and 0.75 micromol x L(-1) for SKOV3 cells, respectively. Further, the enhanced cytotoxicity reduced greatly with the addition of free folic acid.

CONCLUSIONFolate conjugated PGA did not significantly compromise PGA catalytic activity and enabled binding prodrug-activating enzyme PGA to folate receptor expressing cells, and increased the sensitivity of the cells to doxorubicin followed by administration of its prodrug substrate.

Antibiotics, Antineoplastic ; pharmacology ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Doxorubicin ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Female ; Folate Receptors, GPI-Anchored ; Folic Acid ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Inhibitory Concentration 50 ; Ovarian Neoplasms ; pathology ; Penicillin Amidase ; chemistry ; pharmacology ; Prodrugs ; pharmacology ; Receptors, Cell Surface ; metabolism

OBJECTIVETo investigate the protective immunity against Echinococcus granulosus in mice immunized with rEg14-3-3.

METHODSICR mice were subcutaneously immunized three times with rEg14-3-3, followed by the challenge with Echinococcus granulosus protoscoleces intraperitoneally and then sacrificed after six months of post-challenge to detect the proliferation of splenocytes by MTT assay, and to measure the secretion of IL-2, IL-4, IL-10, and IFN-γ by ELISA. The rate of reduced hydatid cyst and the levels of IgE, IgG and IgG subclasses in sera were examined.

RESULTSMice vaccinated with rEg14-3-3 and challenged with protoscoleces revealed significant protective immunity of 84.47%. ELISA analysis indicated that the immunized mice generated specific high levels of IgG and the prevailing isotypes of IgG were IgG1 and IgG2a. Splenocytes from mice immunized with rEg14-3-3 showed a significant proliferation response. The secretion of IFN-γ and IL-2 increased significantly in the vaccinated mice whereas there was no significant difference in IL-4 and IL-10 levels between vaccinated and control mice.

CONCLUSIONThe results indicate that the rEg14-3-3 vaccine could induce a high level of protective immunity as a promising vaccine candidate to prevent cystic echinococcosis.

14-3-3 Proteins ; genetics ; metabolism ; Animals ; Blotting, Western ; Cell Proliferation ; Cytokines ; genetics ; metabolism ; Echinococcosis ; prevention & control ; Echinococcus granulosus ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation ; physiology ; Mice ; Spleen ; cytology ; Vaccines ; immunology

AIMTo construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting TGF-beta1 for further research on the effects of TGF-beta1 on vasculogenesis and angiogenesis.

METHODSThree pairs of siRNA target sequences coding from the mRNA of TGF-beta1 gene were designed and three pairs of nucleotides were synthesized. After annealing, the double-strand DNA products were ligated into the pEN_mH1c entry vector, and in turn into the shRNA eukaryotic expression vector pDS_hpEy labled by GFP through the LR recombination reaction. After sequencing successfully, the three resulting TGF-beta1 shRNA expression vectors were transfected into the mouse fibroblast cell line (NIH/3T3), and then cell clones stably expressing TGF-beta1 shRNA were screened. Reverse Transcript-Polymerase Chain Reaction (RT-PCR) and Western blot were used to detect the mRNA and protein expression.

RESULTSRT-PCR and Western blot showed that one of the TGF-beta1 shRNA expression vectors pDS_Tc downregulated TGF-pl mRNA and protein expression markedly in NIH/3T3 cells.

CONCLUSIONShRNA eukaryotic expression vectors targeting TGF-beta1 are successfully constructed which can be used for further investigation on the mechanism through which TGF-beta1 regulates vasculogenesis and angiogenesis.

Animals ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; Neovascularization, Physiologic ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Transforming Growth Factor beta1 ; genetics ; metabolism

The paper reports the systematic study on felodipine and its impurities in tablets, to improve its quality standards for the control of the related substances. HPLC-DAD, UPLC-MS, IR and NMR methods were used for the isolation of felodipine and its impurities in tablets, their identification and the zebrafish animal model was used for the analysis of the toxic impurities. In felodipine material and its tablets, three impurities are isolated and identified. They are impurity 1 [dimethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], impurity 2 [ethyl methyl 4-(2, 3-dichlorophenyl)-2, 6-dimethylpyridine-3, 5-dicarboxylate] and impurity 3 [diethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], separately. The result of zebrafish animal model analysis showed that the teratogenic effects of four compounds were: impurity 3 > or = felodipine > impurity 1 > impurity 2, lethal effects were as follows: impurity 2 = impurity 3 > felodipine > or = impurity 1. This study confirmed the toxicity of three impurities in felodipine. According to the results, the paper suggested the amendments to the standard of the medicine and provided the support to the control of impurities in the manufacturing process.
Abnormalities, Drug-Induced ; Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; toxicity ; Calcium Channel Blockers ; administration & dosage ; chemistry ; toxicity ; Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Felodipine ; administration & dosage ; chemistry ; toxicity ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Pharmaceutical Preparations ; analysis ; chemistry ; Quality Control ; Spectrophotometry, Infrared ; Tablets ; Tandem Mass Spectrometry ; Zebrafish

OBJECTIVETo investigate the effects of murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of yolk sac hematopoietic progenitors.

METHODSThe serum-free mBMEC-CM was obtained from subcultures of murine endothelial cell line derived from bone marrow which was established in our laboratory. The murine yolk sacs were harvested on day 8.5 postcoitus (pc) and incubated with 0.1% collagenase in 10% fetal calf serum at 37 degrees C for 40 minutes. Yolk sac cells were incubated in tissue culture dishes at 37 degrees C for 1 hour. Nonadherent cells were collected for semisolid culture assay of granulocyte-macrophage colony forming unit (CFU-GM) and high proliferative potential-colony forming cell (HPP-CFC) after being cultured in DMEM with 10% mBMEC-CM and 10% FBS for 24 hours. The number of CFU-GM and HPP-CFC was counted at day 7 and day 14 respectively.

RESULTSThe growth of CFU-GM and HPP-CFC was supported by mBMEC-CM with GM-CSF. mBMEC-CM could induce the proliferation and differentiation of yolk sac hematopoietic stem cells and progenitors in liquid culture system. The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (119.5 +/- 5.7)% and (130.8 +/- 9.8)% respectively after 24 hours liquid culture (P < 0.05). The expansion effects of mBMEC-CM on CFU-GM and HPP-CFC were enhanced by compounded with flt3 ligand (FL) and thrombopoietin (TPO). The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (132.0 +/- 6.2)% and (176.9 +/- 12.8)% respectively after 24 hours liquid culture (P < 0.01).

CONCLUSIONMurine bone marrow endothelial cell conditioned medium could support the growth and proliferation of yolk sac hematopoitic stem cells and progenitors, and this promoting effect was further enhanced by addition of FL and TPO.

Animals ; Bone Marrow Cells ; cytology ; Cell Division ; Cells, Cultured ; Culture Media, Serum-Free ; Endothelium ; cytology ; Female ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Yolk Sac ; cytology

Objective To explore factors influencing the incidence of carotid atherosclerosis in different genders so as to provide reference for the specific prevention of the disease. Methods A nested case-control study was conducted to analyze factors influencing the incidence of carotid atherosclerosis in Jinchang cohort population who were randomly selected through stratified sampling by age and followed up. A risk prediction model was established and the goodness of fit was evaluated by the area under the receiver operator characteristic curve (ROC). Results The standardized incidence of carotid atherosclerosis in this follow-up population was 12.32%, and the incidence rate of males (13.65%) was greater than that of females (11.29%). The difference was statistically significant ( 2=4.267, P<0.001). Age, education, elevated systolic blood pressure, and elevated low-density lipoprotein cholesterol were common risk factors for carotid atherosclerosis in both men and women. Elevated fasting plasma glucose (OR=2.556, 95% CI: 1.618-4.038) and elevated triglyceride (OR=1.535, 95% CI: 1.058-2.227) were only associated with men. Abdominal obesity (OR=1.414, 95% CI: 1.013-1.974) was only associated with women. The area under ROC of male and female prediction models was 0.835 (95% CI: 0.815-0.856) and 0.809 (95% CI: 0.788-0.831), respectively. The sensitivity was 78.0% and 78.9%, the specificity was 78.8% and 73.1%, and the diagnostic coincidence rate was 91.3% and 82.4%, respectively. Conclusions There are different risk factors for carotid atherosclerosis in males and females, and targeted prevention and control measures should be taken according to gender. The risk prediction model established by Logistic regression had certain guiding value.

Toxoplasma gondii is a serious foodborne zoonosis,which can not only affect the development of animal husbandry and the safety of meat products,but also cause great harm to public health.Therefore,the prevention of toxoplasmosis is crucial.Toxoplasma gondii vaccine which is regarded as an important measure to prevent toxoplasmosis has significant value to both public health and economics.This paper mainly summarizes advances in the study of Toxoplasma gondii vaccine in order to provide references for the further development of it,so as to control the toxoplasmosis more effectively.