Urokinase plasminogen activator receptor (uPAR) is a membrane protein which is attached to the cellular external membrane. The uPAR expression can be observed both in tumor cells and in tumor-associated stromal cells. Thus, in the present study, the human amino-terminal fragment (hATF), as a targeting element to uPAR, is used to conjugate to the surface of superparamagnetic iron nanoparticle (SPIO). Flowcytometry was used to examine the uPAR expression in different tumor cell lines. The specificity of hATF-SPIO was verified by Prussian blue stain and cell phantom test. The imaging properties of hATF-SPIO were confirmed in vivo magnetic resonance imaging (MRI) of uPAR-elevated colon tumor. Finally, the distribution of hATF-SPIO in tumor tissue was confirmed by pathological staining. Results showed that the three cells in which we screened, presented different expression characteristics, i. e., Hela cells strongly expressed uPAR, HT29 cells moderately expressed uPAR, but Lovo cells didn't express uPAR. In vitro, after incubating with Hela cells, hATF-SPIO could specifically combined to and be subsequently internalized by uPAR positive cells, which could be observed via Prussian blue staining. Meanwhile T2WI signal intensity of Hela cells, after incubation with targeted probe, significantly decreased, and otherwise no obvious changes in Lovo cells both by Prussian blue staining and MRI scans. In vivo, hATF-SPIO could be systematically delivered to HT29 xenograft and accumulated in the tumor tissue which was confirmed by Prussian Blue stain compared to Lovo xenografts. Twenty-four hours after injection of targeting probe, the signal intensity of HT29 xenografts was lower than Lovo ones which was statistically significant. This targeting nanoparticles enabled not only in vitro specifically combining to uPAR positive cells but also in vivo imaging of uPAR moderately elevated colon cancer lesions.
Cell Line, Tumor ; Colonic Neoplasms ; diagnosis ; Ferric Compounds ; Humans ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; chemistry ; Molecular Imaging ; methods ; Receptors, Urokinase Plasminogen Activator ; chemistry ; Staining and Labeling

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Objective To investigate the effects of exercise training(ET)on the expression of glucogen synthase kinase 3 bate(GSK 3?)in the adipose tissues of insulin resistant(IR)rats on a high fat diet(HFD). Methods Thirty male Wistar rats were randomly divided into a control group(n=10)and a model group(M group,n=20).Insulin resistance was established by feeding a HFD to the M group for 4 w,while rats in the control group were fed a normal diet.The IR rats were then randomly divided into two subgroups:an IR group and an ET group.All rats in the IR and ET groups were fed a HFD,but ET was administrated to the ET group for 6w.The expres- sion of GSK 3?protein in the rats'epididymis adipose tissue was detected using Western blotting,and body weight (BW),the concentrations of serum triglyceride and cholesterol(TG and TC),fasting plasma glucose(FPG)and serum insulin(FINS),as well as insulin sensitivity index(ISI)were regularly detected.Results Compared with the con- trol group,BW and the concentrations of serum TG and TC,FPG and FINS in the model group were significantly in- creased(P＜0.05),while ISI was decreased(P＜0.01).Compared with the control group,there was no difference in GSK 3 protein expression in the ET group,but the expression of GSK 3?protein in the ET group was obviously de- creased in comparison with that in the IR group(P＜0.05).Conclusion ET can ameliorate IR by decreasing GSK 3?protein expression in adipose tissues and enhancing the ingestion of glucose and the synthesis of glycogen.

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

An enhancer-like element VV16 from Vaccinia virus genome DNA was obtained by using the plasmid with CAT reporter gene. Sequence analysis showed the element of 112bp is a part of the DNA-dependent RNA polymerase, polyA polymerase and DNA polymerase (RPO30 gene). It contains 4 AT-rich regions. Detection of β-galactosidase activity showed that VV16 in the positive direction can increase the activity 9.0 times and VV16 in the negative direction can increase 4.1 times. The RNA dot blotting confirmed the enhancing activity of the element are on the transcription level. DNA deletion experiment indicated the sequences of 10bp at the 5′ end and 12bp at the 3′ end in the element are important to its function and the sequence from nt76 to nt82 is essential to its activity.

OBJECTIVETo observe three-dimensional space position change of nucleus pulposus and nerve root before and after treatment of lumbar disc herniation by spinal fixed-point rotating reduction, and explore the mechanisms.

METHODSTotally 52 patients with L5S1 lumbar disc hernation treated by spinal fixed-point rotating reduction were collected from April 2009 to June 2011. There were 33 males and 19 females with an average age of 34.6 years old (ranged, 19 to 55). Three-dimensional MRI were performed to observe relationship between nucleus pulposus and related nerve root,configuration change of spine and pelvic on coronary MRI.

RESULTSMRI showed relationship between nucleus pulposus and related nerve root mainly located on axillary, shoulder, front and surround. Vertebral displacement disappeared, lumbocrural pain alleviated after manipulative therapy. All patients were followed up from 2 to 28 months with an average of 12 months, and no recurred. All patients recovered work. Nucleus pulposus had no change,while lumbral spinal and pelvic curve changed before and after admission.

CONCLUSIONLumbar disc herniation combined with single (multiple) vertebral displacement,can cause biomechanical properties of nucleus puplosus and related nerve root, while spinal fixed-point rotating reduction can correct vertebral displacement, recover balance between inside and outside of spinal.

Adult ; Female ; Humans ; Imaging, Three-Dimensional ; Intervertebral Disc Displacement ; diagnosis ; diagnostic imaging ; physiopathology ; therapy ; Lumbar Vertebrae ; diagnostic imaging ; physiopathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Rotation ; Young Adult

OBJECTIVETo evaluate the clinical effect of the treatment for sequestered type lumbar disk herniation (SLDH) with FENG's spinal manipulation.

METHODSForty-one patients (27 male, 14 female) were treated by FENG's spinal manipulation. The clinical results were measured by the Japanese and FENG's lumbar treatment standard and evaluated by scoring system for assessment of treatment for low back pain between pre- and post-treatment. To measure MRI sagittal index (SI) of lumbar disc herniation between pre- and post-treatment.

RESULTSThe score of subjective symptoms and clinical signs increased obviously after treatment. There was no statistical difference of SI between pre- and post-treatment (P<0.05). The outcome of treatment was satisfying.

CONCLUSIONThe major pathologic change of SLDH is the single/multiple vertebral subluxation. FENG's spinal manipulation can correct the vertebral subluxation which leading to the series of clinical manifestations. The good clinical outcome may due to the alleviation of distortion and tension of dura and nerve root with the recovery of the spinal column's stability.

Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Manipulation, Spinal ; methods ; Middle Aged

OBJECTIVETo study the compatibility of composite herbal medicines of the Ling Gui Zhu Gan Decoction.

METHODEthanol extract test solutions of the different combinations were prepared according to the orthogonal layout L16(4(5)). Pharmacologic experiments, such as the time of surviving of mice in shortage of oxygen in regular air pressure, the antagonizing effect on arrhythmia induced by chloroform and diuresis were carried out with the solutions. Variance analysis, canonical correlation and stepwise regression analysis were applied to interrelate the amount of each drug and the pharmacologic data.

RESULTThe results confirmed that Fuling and Guizhi are the basis, while Baizhuand Gancao are the adjuvans, which is conformed to the theory of TCM.

CONCLUSIONThis study provides a significant try for studying the compatibility of composite herbal medicines.

Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Diuretics ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Female ; Hypoxia ; drug therapy ; Male ; Medicine, Chinese Traditional ; Mice ; Phytotherapy ; Plant Extracts ; isolation & purification ; therapeutic use ; Plants, Medicinal ; chemistry ; Ventricular Fibrillation ; drug therapy

OBJECTIVETo screen enhancer-like sequences from vaccinia virus genome, to construct an expression vector harboring prokaryotic enhancer-like sequence and study the effect of interferon gene expression.

METHODSEnhancer-like element from vaccinia virus genome was obtained by using the chloramphenicol acetyl-transferase cat gene as reporter gene. An expression vector harboring prokaryotic enhancer-like sequence VV1 from vaccinia virus was constructed. Interferon was expressed and assayed.

RESULTSEighteen enhancing sequences were found. From them two enhancer-like sequences with distance and orientation independence property were screened and named VV1 and VV16 respectively. Quantification test showed that the direct and reverse orientation of VV1 could increase the activity of beta-galactosidase with 10.9 and 3.8 times and those of VV16 could increase by 9.0 times and 4.1 times respectively. The enhancing activity of the element was on transcription level. An expression vector harboring prokaryotic enhancer-like sequence VV1 was constructed. Using this vector the antiviral activity of interferon alpha-2b was increased by 2.6 times in comparison with the original expression plasmid.

CONCLUSIONTwo enhancer-like sequences were screened from vaccinia virus genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.

Enhancer Elements, Genetic ; Genetic Vectors ; genetics ; Interferon-alpha ; biosynthesis ; genetics ; pharmacology ; Plasmids ; Recombinant Proteins ; Vaccinia virus ; genetics

Objective To discuss the status of self-care ability and health behavior in gynecologic patients receiving chemotherapy,and analyze their correlation.Methods ESCA and HPL were used to investigate 90 cases of gynecologic patients receiving chemotherapy and analyze the correlation.Results The score of self-care ability was (112.17 ± 17.46),health behavior was (123.21 ± 20.23),and there was a positive correlation between the self-care ability and health behavior in gynecologic patients receiving chemotherapy (r =0.275-0.705,P ＜ 0.05).Conclusions Nurses should take effective nursing measures to improve self-care ability of gynecologic patients receiving chemotherapy,so as to promote them take health behaviors to reduce tumor recurrence and improve cancer survivors' quality of life.