Agmatine ; administration & dosage ; pharmacology ; Analgesia ; methods ; Animals ; Injections, Spinal ; Male ; Morphine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; drug effects

AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

OBJECTIVETo compare the differences in uptake of 2-deoxy-D-glucose (2-DG)-conjugated nanoparticles between breast carcinoma MDA-MB-231 cells with high metabolism and breast fibroblasts with normal metabolism, and investigate the feasibility of using the coated nanoparticles as a MRI-targeted contrast agent for highly metabolic carcinoma cells.

METHODSThe γ-Fe2O3@DMSA-DG was prepared. The glucose metabolism level of both cell lines was determined. The targeting efficacy of γ-Fe2O3@DMSA-DG and γ-Fe2O3@DMSA NPs to breast carcinoma MDA-MB-231 cells and breast fibroblasts at 10 min, 30 min, 1 h and 2 h was measured with Prussian blue staining and UV colorimetric assay. MRI was performed to visualize the changes of T2WI signal intensity.

RESULTSPrussian blue staining showed more intracellular blue granules in the MDA-MB-231 cells of γ-Fe2O3@DMSA-DG NPs group than that in the γ-Fe2O3@DMSA NPs group, and the γ-Fe2O3@DMSA-DG uptake was greatly competed by free D-glucose. As revealed by UV colorimetric assay, MDA-MB-231 cells also showed that the cellular iron amount of γ-Fe2O3@DMSA-DG group was significantly higher than that of the γ-Fe2O3@DMSA group and γ-Fe2O3@DMSA-DG + D-glucose group, statistically with a significant difference between them. MRI showed that the signal intensity of γ-Fe2O3@DMSA-DG group was decrease significantly, the T2 signal intensity was decreased by 10.5%, 37.5%, 72.9%, 92.0% for 10 min, 30 min, 1 h and 2 h, respectively. In contrast, the signal intensity did not show obvious decrease in the γ-Fe2O3@DMSA-DG group, the T2 signal intensity was decreased by 8.5%, 11.4%, 32.0%, 76.7% for 10 min, 30 min, 1 h and 2 h, respectively. However, HUM-CELL-0056 cells did not produce apparent difference for positive staining in the γ-Fe2O3@DMSA-DG group, γ-Fe2O3@DMSA group and γ-Fe2O3@DMSA-DG+D-glucose group, and the signal intensity also did not produce apparent difference.

CONCLUSIONSγ-Fe2O3@DMSA-DG has good targeting ability to highly metabolic breast carcinoma (MDA-MB-231) cells. It is feasible to serve as a specific MRI-targeted contrast agent for highly metabolic carcinoma cells, and deserves further studies in vivo.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Colorimetry ; methods ; Contrast Media ; pharmacokinetics ; Deoxyglucose ; chemistry ; pharmacokinetics ; Female ; Ferric Compounds ; chemistry ; pharmacokinetics ; Fibroblasts ; cytology ; metabolism ; Glucose ; metabolism ; Humans ; Iron ; metabolism ; Magnetic Resonance Imaging ; methods ; Nanoconjugates ; chemistry ; Particle Size ; Succimer ; chemistry ; pharmacokinetics

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.
Animals ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Cell Nucleus ; metabolism ; Cerebral Cortex ; metabolism ; Female ; Fibroblast Growth Factor 1 ; administration & dosage ; pharmacokinetics ; Gene Products, tat ; administration & dosage ; pharmacokinetics ; Hippocampus ; metabolism ; Injections, Intravenous ; Male ; Mice ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; administration & dosage ; pharmacokinetics

OBJECTIVETo assess the distribution frequency of Gly82Ser polymorphism of receptor for advanced glycation end products (RAGR) gene and investigate its association with type 2 diabetic Chinese patients.

METHODSThe allele frequencies and genotype distribution of Gly82Ser polymorphism of RAGE gene were compared in a case-control study of 194 type 2 diabetic and 546 non-diabetic subjects. PCR-restriction fragment length polymorphism (PCR-RFLP) was used for detection of the genotype variants.

RESULTSIn general Chinese population and type 2 diabetic Chinese patients, the most frequent genotype and allele of RAGR gene Gly82Ser polymorphism were genotype GG and allele G, whose frequency distribution were significantly higher than those in other countries (P<0.01). No significantly difference in the genotype frequencies or allele frequencies of Gly82Ser polymorphism were found between the diabetic patients and non-diabetic subjects (P>0.05).

CONCLUSIONGly82Ser polymorphism of RAGE gene does not demonstrate any association with type 2 diabetes in Chinese patients, but high genotype and allele frequencies of Gly82Ser polymorphism occur in Chinese population and type 2 diabetic Chinese patients.

Aged ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; China ; Diabetes Mellitus, Type 2 ; ethnology ; genetics ; Female ; Gene Frequency ; Genotype ; Glycine ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; genetics ; Serine ; genetics

OBJECTIVETo observe the role of magnesium sulfate in vascular calcification, to explore the role and the mechanism of magnesium sulfate in vascular calcification.

METHODSThe vascular calcification model was established by administration of vitamin D3 plus nicotine (VDN) in SD rats. To estimate the extent of calcification by Von Kossa staining, calcium content and alkaline phosphatase activity, osteopontin (OPN) mRNA were determined by using semi-quantitative reverse-transcription polymerase chain reaction.The malondialdehyde (MDA) and nitric oxide (NO) content and activities of superoxide dismutase(SOD) were measured by biochemistry.

RESULTSA strong positive staining of black/brown areas among the elastic fibers of the medial layer in calcified aorta by Von Kossa staining, calcium content and ALP activity in calcified arteries increased by 3.9-and 3.4-fold as compared with the controls. The expression of OPN mRNA was up-regulated by 40% (P < 0.01). The lipid peroxidation products MDA in vascular were increased 2.0-fold (P < 0.01). The NO content and SOD activity were greatly decreased by 64% and 72% (P < 0.01), respectively, compared with controls. However, calcium content and ALP activity in VDN plus magnesium sulfate group were lower than those in VDN group. Low and high dosage magnesium sulfate obviously relieved degree of calcification in the cardiovascular tissues in a dosage-dependent manner (P < 0.01).

CONCLUSIONMagnesium sulfate plays a role in the pathogenesis of vascular calcification by reducing vascular calcification and decreasing vascular injury.

Animals ; Cholecalciferol ; adverse effects ; Magnesium ; pharmacology ; Male ; Nicotine ; adverse effects ; Osteopontin ; metabolism ; RNA, Messenger ; genetics ; Rats ; Vascular Calcification ; chemically induced ; pathology

OBJECTIVETo investigate the role of the expression of ephrinB2 and EphB4 in non-small cell lung cancer (NSCLC), and their relationship with multi-slice spiral CT pulmonary perfusion imaging.

METHODSThirty-one nodules with pathologically proven NSCLC underwent CT perfusion scan. The perfusion parameters including blood flow (BF), blood volume (BV), peak enhancement image (PEI) were collected. The expression of ephrinB2 and EphB4 in tumor cells and interstitial vasculature were detected by immunohistochemistry. Correlation analysis and trend test were used to assess the relationship between ephrinB2/EphB4 expression and clinicopathological features, and between ephrinB2/EphB4 expression and perfusion parameters.

RESULTSPositive expression of ephrinB2 and EphB4 in the NSCLC group was 83.9% and 71.0%, respectively, significantly higher than that in the internal control group (P < 0.01). The expression of ephrinB2 and EphB4 was consistently in tumor parenchyma but differently in tumor vessels. The expressions of ephrinB2 and EphB4 were positively correlated with lymphatic metastasis (P < 0.05). The expression of EphB4 was negatively correlated with blood flow (BF) and blood volume (BV), respectively (P < 0.05). There was a significant positive correlation between ephrinB2 expression and BF (r = 0.516, P = 0.003), and a positive correlation between ephrinB2 expression and BV (r = 0.448, P = 0.013). The expressions of ephrinB2 and EphB4 were not correlated with PEI (P > 0.05). The values of BF and BV in the high and moderate EphB4 expression groups were significantly decreased compared with that in the negative group (P < 0.01). The value of BF in the high ephrinB2 expression group was significantly increased compared with that in the moderately positive group and negative group (P < 0.01). The value of BV in the high ephrinB2 expression group was significantly increased compared with that in the negative group (P < 0.01).

CONCLUSIONThe CT pulmonary perfusion imaging reflects the density difference of blood vessels with functional lumen, and such difference also depends on the quantity and quality of vasculature with functional lumen.

Adult ; Aged ; Blood Volume ; Carcinoma, Non-Small-Cell Lung ; blood supply ; diagnostic imaging ; metabolism ; pathology ; physiopathology ; Ephrin-B2 ; metabolism ; Female ; Humans ; Lung Neoplasms ; blood supply ; diagnostic imaging ; metabolism ; pathology ; physiopathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Perfusion Imaging ; Pulmonary Circulation ; Receptor, EphB4 ; metabolism ; Tomography, Spiral Computed ; methods

Objective To learn the current neglected situation and its impact factors on rural children in two provinces in Western China.Methods The investigation was conducted by using multistage stratified cluster sampling method in some parts of the Shaanxi province and the city of Chongqing.Results 1488 subjects,aged from 0 to 6,were recruited in the present study.Results showed that the total prevalence rates of neglected rural children in the two research sites were 31.59％ and 48.32 respectively.No significant difference was found on the prevalence of neglected for boys and girls ( x2 =0.86,U=1.51,P ＞ 0.05 ).The degrees of negligence in the older children showed a significant increase than in the younger children (x2 =13.36,F=33.45,P＜ 0.05 ).The prevalence and degree of negligence in families with three generations were lower than that of a single-parent families and remarried families (H=10.03,F=2.83,P＜0.05).The one-child-families were significantly lower than multiple children families,both in terms of prevalence and degree of negligence.Our data demonstrated the degrees of negligence in children whose parents were away from home,were higher than the children whose parents stayed with them ( x2=30.30,U=6.76,P＜0.05).The degree of negligence in Shaanxi was lower than that in Chongqing (x2=9.42,U=8.40,P＜0.05).Based on multivariate non-conditional logistic regression analysis,the determinants of stunting among the stranded children were as follows:parents were away from home (OR=1.54,95％ CI:1.20-1.97) ;structure of the family (OR=1.38,95％ CI:1.16-1.65) ; father' s occupation (OR=0.87,95％ CI:0.78-0.97) ; schooling of the children' s mother (OR=1.27,95％CI:1.07-1.52) ; relationship between children and their fathers (OR =1.43,95％ CI:1.07-1.91 ) etc.Conclusion The situation of negligence in children living in the rural areas,were serious in Shaanxi and Chongqing provinces,which called for the strengthening on the publicity and education of the issue.School and the society should also pay more attention to this problem.

Objective To investigate the value of endoscopy combined with laparoscopy in the treatment of colorectal polyps and polyp canceration.Methods Different combinations of endoscopic and laparoscopic procedures were employed and the clinical efficacies were compared.Results From January 2004 to September 2006,46 cases with colorectal polyp were treated with endoscopy combined with laparoscopy.Among them,5 cases(10.87%)underwent laparoscopic-assisted endoscopic polypectomy,30(65.22%)endoscopic-assisted laparoscopic resection,6(13.04%)synchronously endoscopic and laparoscopic resection.Five cases were performed further operation after endoscopic polypectomy.According to the pathological findings,21(45.7%)were proved to be polyp canceration,among which 6 were advanced carcinoma,and 3 were found metastasis to the lymph nodes.Among the 41 cases of laparoscopic resection,there was no conversion to an open surgery.Anastomotic leakage was found in 2 cases and anastomotic bleeding in 1.In the 5 cases of laparoscopic-assisted endoscopic polypectomy,no complication was observed.During the period of follow-up(1 to 21 months),no recurrence was detected.Conclusion Endoscopy combined with laparoscopy extends the safety and indications of endoscopic polypectomy,and is minimally invasive to the patients.It is an ideal procedure in the treatment of colorectal polyps and poly carceration.

OBJECTIVETo investigate the effect of bone marrow stem cell transplantation (BMT) on the diaphragm muscles of mdx mice, a mouse model of Duchenne muscular dystrophy (DMD).

METHODSThe bone marrow-derived stem cells form male SD rats was transplanted through the tail vein into 18 female 8-week-old mdx mice, which were sacrificed at 4, 8 and 12 weeks after BMT (6 at each time point), respectively. The diaphragm muscles of the mice were subjected to HE staining, immunofluorescence detection of dystrophin, reverse transcription (RT)-PCR analysis of dystrophin mRNA transcripts and PCR analysis of Sry (sex-determining region on the Y chromosome) gene, with age-matched female C57 mice and untreated mdx mice as the controls.

RESULTSThe proportion of centrally nucleated fibers (CNF) in the diaphragm muscle of the recipient mdx mice was (15.58+/-0.91) %, (12.50+/-1.87) % and (10.17+/-1.17) % at 4, 8 and 12 weeks after BMT, respectively, significantly smaller than that of untreated mdx mice [(19.5+/-1.87) %], and the fibers after BMT showed less inflammatory infiltration. Compared with the untreated mice, the recipient mdx mice showed green fluorescence on significantly more diaphragm muscle cell membranes [with the proportion of dystrophin-positive fibers of (1.00+/-0.32) %, (6.00+/-1.05) % and (11.92+/-1.11) % at 4, 8, and 12 weeks after BMT]. RT-PCR of dystrophin mRNA also demonstrated significantly higher relative levels of dystrophin in the recipient mdx mice (0.19+/-0.05, 0.26+/-0.06 and 0.36+/-0.04 at 4, 8 and 12 weeks after BMT) than in untreated mdx mice, and Sry gene was present in the recipient mice.

CONCLUSIONBMT can partially restore dystrophin expression and ameliorate the pathology in the diaphragm muscles of mdx mice, and has great potential to produce general therapeutic effect in patients with DMD.

Animals ; Bone Marrow Transplantation ; methods ; Diaphragm ; metabolism ; pathology ; Dystrophin ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred mdx ; Muscular Dystrophy, Duchenne ; metabolism ; pathology ; surgery ; Rats ; Rats, Sprague-Dawley ; Transplantation, Heterologous