The PET tracer 5-([11C]methyloxy)-L-tryptophan (5-(11)CMTP) was prepared by nucleophilic fluorination and alkylation reaction via a two-step procedure in order to develop specific tumor probe. The biodistribution and microPET imaging of 5-(11)CMTP were executed. The results unveiled that the overall radiochemical yield with no decay correction was (14.6 ±7.2) %, the radiochemical purity was more than 95% and high uptake and long retention time of 5-(11)CMTP in liver, kidney and blood were observed but low uptake in brain and muscle were found, furthermore, high uptake of 5-(11)CMTP in tumor tissue was observed. It seems that 5-(11)CMTP will be a potential amino acid tracer for tumors imaging with PET.
Amino Acids ; Animals ; Neoplasms ; diagnostic imaging ; Positron-Emission Tomography ; Radioactive Tracers ; Tissue Distribution ; Tryptophan ; analogs & derivatives

Objective To explore the relationship between the inflammatory reaction and insulin function in children with critically ill.Me-thods Ninty-six children with critical disease in Oct.2003 to Oct.2006 were enrolled in the study.Blood sugar,plasma insulin,C-peptide,tumor necrosis factor(TNF)-?,C reactive protein(CRP)were measured in the peak period and convalescence.Results Blood sugar and plasma levels of insulin,C-peptide,TNF-?,CRP were significantly higher in the peak period than those in the convalescence(Pa

OBJECTIVETo investigate the effect of Shuizhongcao Granule (SZCG) on cellular immune function in animal model of recurrent aphthous stomatitis (RAS).

METHODSExperimental animal model of RAS were established by immunological method. The modeled animals were randomized into 4 groups, Group A was the model control group, Group B was treated by Levamisole (2.5 mg/mL), Group C and D was treated with high-dose (0.5 g/mL) and low-dose (0.24 g/mL) SZCG respectively. The medication was administered via gastric perfusion, starting from the 7th week of the experiment, and continued for 28 days. Meanwhile, a normal control group was set up. Percentage of T-lymphocyte subsets, CD3, CD4 and CD8, and serum levels of interleukin 10 and 12 (IL-10 and IL-12) in peripheral blood were detected before and after treatment.

RESULTSPercentages of CD3, CD4 and CD4/CD8 ratio in the model control group were 40.50 +/- 7.46%, 30.80 +/- 5.33% and 79.56 +/- 8.32 respectively, all lower than those in the normal control group (P<0.05); level of serum IL-10 was higher (8.02 +/- 0.53 ng/L) and that of IL-12 was lower (15.51 +/- 1.35 ng/L) in the model animal than those in normal control (P<0.05). After treatment, levels of CD3, CD4, CD4/CD8 and IL-12 in Group B were 55.30 +/- 6.33%, 43.50 +/- 5.33%, 99.29 +/- 13.84 and 17.72 +/- 1.70 ng/L, those in Group C were 61.50 +/- 4.32%, 47.20 +/- 4.37%, 103.30 +/- 7.42 and 18.18 +/- 1.54 ng/L, and in Group D were 58.50 +/- 6.03%, 42.80 +/- 3.17%, 110.27 +/- 12.85 and 17.74 +/- 1.96 ng/L, respectively, all were higher than those in Group A, but levels of IL-10 in the three treatment groups were lower than that in Goup A (P<0.05 or P<0.01).

CONCLUSIONThe immune function of RAS model animals is in the suppressed condition; SZCG can improve the immune suppression to achieve its therapeutic effect on the disease.

Animals ; CD4-CD8 Ratio ; Drugs, Chinese Herbal ; therapeutic use ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Male ; Mouth Mucosa ; Phytotherapy ; Rabbits ; Stomatitis, Aphthous ; drug therapy ; immunology ; T-Lymphocyte Subsets ; immunology

A sensitive electrochemical sensor for the detection of semicarbazide ( SEM ) based on the glassy carbon electrode modified with carboxylated multi-walled carbon nanotube ( CMWCNTs ) was constructed in this work. The modified materials were characterized by Fourier transform infrared spectroscopy, transmission electron microscopy and electrochemical impedance spectroscopy. The results showed that the CMWCNTs exhibited a certain degree of changes, such as the appearance of the characteristic peaks of carbon oxygen double bonds in carboxyl group, a decrease in diameter and length and a remarkable reduction of the impedance value. The electrochemical behavior of SEM on CMWCNTs/GCE was investigated by cyclic voltammetry and amperometric i-t curve in 1 mol/L HAc-NaAc buffer solution. An irreversible oxidation peak of SEM appeared at CMWCNTs/GCE. Compared to the bare electrode, the current of the oxidation peak was significantly increased. Under the optimal conditions such as HAc-NaAc solution ( pH 7 ) and scan rate of 0. 1 V/s, the obtained sensor presented linear response to SEM concentration in the range of 5. 0 × 10-6 mol/L-1. 09×10-3 mol/L, the linear regression equation was Ip(μA)=-0. 472+0. 0599C (μmol/L) with linear correlation coefficient of 0. 997. The detection limit was 1. 88×10-7 mol/L. At the same time, the recovery was from 92. 8% to 98. 0% in the test of pork liver samples.

Xanthomonas campestris pv. campestris ( Xcc), causative agent of the black rot disease of cruciferous crops worldwide, produces large amount of extracellular polysaccharide( EPS), which has found wide applications in industry. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5gus A5, and a number of EPS-defective mutants were isolated. The Tn5gusA5 insertion sites in the mutants were analyzed by using thermal asymmetric interlaced PCR(TAIL-PCR), and the corresponding genes were identified by homology blast to the completely sequenced genome of Xcc 8004 strain. A novel gene, waxE, identified from the EPS-defective mutant 151D09, was found to be disrupted by the insertion of Tn5gusA5 in the open reading frame(ORF) with genome coordinates 4478998bp to 4479819bp.This gene showed 52% similarity to the kdtX gene of Serratia marcescens and 50% to the waaE of Klebsiella pneumoniae at amino acid level, with characteristics of glycostransferase 2 family domain. In order to identify the function of waxE gene, waxE gene deletion mutant of Xcc 8004 was constructed by gene replacement strategy in which waxE gene of genome was replaced by kanamycin resistant gene kan. The waxE gene deletion mutant strain, named Xcc 8570, was confirmed by both PCR and southern analysis. The growth rate of the deletion mutant 8570 in rich medium was not affected, but the EPS yield reduced by 35% as compared with the wildtype strain 8004. The deletion mutant could be completmented in trans with plasmid pLATC8976 harboring an intact waxE gene, and the EPS yield of the mutant was restored. The combined data showed that waxE gene involved in EPS biosynthesis in Xcc.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; genetics ; metabolism ; Blotting, Southern ; Cloning, Molecular ; DNA Transposable Elements ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polysaccharides, Bacterial ; genetics ; metabolism ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Xanthomonas campestris ; genetics ; metabolism

Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.
Cell Proliferation ; Genes, Bacterial ; physiology ; Lipopolysaccharides ; biosynthesis ; Mutation ; Polysaccharides, Bacterial ; biosynthesis ; Xanthomonas campestris ; genetics

A sensitive antibody-based lateral flow dipstick was developed for ginsenoside Re (GRe) detection. The stick consisted of a sample pad, a conjugate pad, membrane and an absorbent pad. The membrane was coated with two capture reagents, GRe-BSA conjugate and goat anti-mouse antibodies, forming a test line and a control line, respectively. The conjugate pad was saturated with colloidal gold particles coated with affinity purified monoclonal anti-GRe antibody. The visual detection limit was 200 microg x L(-1) of GRe and the reaction time was 10 min. The Panax ginseng roots were identified after these samples (10 mg) were extracted with 5 mL tap water for 30 min at room temperature, and the extracts were tested by the dipsticks and ELISA kit. The true and false P. ginseng could be distinguished with dipsticks. The dipstick could be used to detect the quality of the P. ginseng samples when the extract was diluted 100-folds. The results were compared with those obtained using an indirect competitive enzyme-linked immunosorbent assay (icELISA). The dipstick assay proved to be a sensitive and rapid tool for quality control of P. ginseng.
Animals ; Antibodies, Monoclonal ; immunology ; Counterfeit Drugs ; analysis ; Ginsenosides ; analysis ; Immunoassay ; instrumentation ; methods ; Mice ; Panax ; chemistry ; Reagent Strips ; Time Factors

AIMTo determine the effects of cyclovirobuxine D (CD) on intracellular Ca2+ mobilization and L-type Ca2+ current (I(Ca-L)) in isolated rat cardiomyocytes.

METHODSThe effects of CD on the amplitude of I(Ca-L) and intracellular Ca2+ mobilization induced by KCl and caffeine were studied with the method of patch-clamp technique and laser scanning confocal microscopy in rat ventricular myocytes.

RESULTSCD decreased the amplitude of I(Ca-L) in a concentration-dependent manner. At 10 mV, 1 and 10 micromol x L(-1) CD decreased I(Ca-L) density from (- 9.9 +/- 1.8) pA/pF to (-6.4 +/- 1.4) and (-4.2 +/- 0.6) pA/pF, respectively. Confocal experiments showed that intracellular fluorescent intensity (FI) value of [Ca2+] in control resting level was not changed by 1 and 10 micromol x L(-1) CD. [Ca2+] increase in response to KCl could not be reduced by CD. The rise of [Ca2+]i in response to caffeine was further enhanced by pretreatment with CD.

CONCLUSIONCD decreased I(Ca,L) in a concentration-dependent manner and increased [Ca2+]i release induced by caffeine in rat ventricular cardiomyocytes.

Animals ; Buxus ; chemistry ; Calcium ; metabolism ; Calcium Channels, L-Type ; drug effects ; Cell Separation ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Heart Ventricles ; cytology ; Male ; Myocytes, Cardiac ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

Objective To evaluate whether 18F-fluorothymidine(FLT) can be used to monitor early response to irradiation in colorectal cancer (CRC).Methods SW480 cells were cultured and irradiated with 0, 10, and 20 Gy.Twenty-four hours later, morphological changes, apoptosis, necrosis, proliferation,and cell cycle phases were observed.Uptake of 18F-FLT was measured in these tumors in vitro from 24 h to 72 h after irradiation.The one-way analysis of variance was used to analyze the data.Results Apoptotic and necrotic cells were detected 24 h after radiotherapy.SW480 cells proliferation was significantly delayed after irradiation in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTI) assay.Cell cycle analysis showed that SW480 cells had a decreased fraction of cells in S phase( from 33.23% to 9.24%,then to 5.43% ) and an arrested fraction in G0-G1.After SW480 cells were cultured for60 min, the uptake of 18F-FLT was (5.21 ± 1.60) %; and 24 h after irradiation of 10 Gy, the uptake decreased significantly to (4.27±0.48)% (F=8.253, P=0.009).And 72 h after irradiation, the uptake further decreased significantly to (3.39 ± 0.59) % ( F = 36.715, P＜0.001 ).In tumor tissue, the uptake of 18F-FLT reduced significantly 72 h after radiotherapy (10 Gy:F = 12.388, P = 0.007; 20 Gy:F = 16.744, P = 0.004) and the attenuation degree increased with the radiation dose.Conclusion The uptake of 18F-FLT in SW480 cells or in CRC could reflect the changes of SW480 cells in proliferation, cell cycle re-distribution, cell apoptosis and necrosis.The results suggest that 18F-FLT may be used for monitoring early response to irradiation of CRC.

Objective To evaluate the predictive value of 18F-fluorodeoxyglucose (FDG) and 18F-fluorothymidine(FLT) PET in monitoring the metastatic potential of human colorectal cancer (CRC).Methods Human CRC cell lines SW480 and SW620 were cultured and implanted into nude mice to create CRC models. Tumor growth,metastatic status and survival were assessed in CRC bearing mice. Uptake of 18F-FDG and 18F-FLT in SW480 and SW620 cells was detected In vitro at 0,30,60,90,120 min after incubation. PET images of both tracers were acquired for SW480 and SW620 tumor-bearing mice using the small animal PET at 60 min after tracer injection. Region of interest (ROI) was drawn using Image J software on reconstructed PET images. Immunocytochemistry and Western blot analysis of the tumor tissue were performed. The correlation between tracer uptake and tumor marker expression was evaluated using linear regression. Results Compared with SW620 tumor-bearing mice,SW480 induced tumor grew much faster ( t = - 3.332,P = 0.004),the tumor-bearing mice had more serious dyscrasia ( t = 2.240,P = 0.038 ),shorter survival and higher metastatic rate. In vitro study,the uptake of both 18F-FDG and 18F-FLT in SW620 cells was lower than that in SW480 cells. 18F-FLT uptake was higher than 18F-FDG uptake in both SW480 and SW620 cells. After incubation for 60 min,the uptake of 18F-FDG in SW480 and SW620 cells was ( 1.76 ± 0.87 )% and ( 1.14 ± 0.38 )%,respectively ( t = - 2.507,P = 0.021 ); while the uptake of 18F-FLT in SW480 and SW620 cells was (5.21 ± 1.60)% and (2.90 ± 1.82)%,respectively (t =3.497,P =0.002). In micro-PET study,the 18F-FDG radioactivity ratio of tumor to non-tumor (T/NT) in SW480 and SW620 tumors was 2.69 ± 0.98 and 3.09 ± 1.26 respectively (t =0.657,P =0.524); while T/NT of 18F-FLT in SW480 and SW620 tumors was 3.65 ±0.51 and 2.22 ±0.42 (t =6.491,P ＜0.001 ),respectively. In immunocytochemistry and western blot assay,heat shock protein(HSP) 27,Integrin β3,vascular endothelial growth factor receptor 2 ( VEGFR2 ) and Ki67 were all over expressed in two kinds of tumor cells with different intensities. HSP27 and Integrin β3 expression was higher in SW480 cells than that in SW620 cells. While VEGFR and Ki67 expression was lower in SW480 cells than that in SW620 cells. The uptake of 18F-FLT closely correlated with the expression of HSP27 ( r =0.924,P =0.004) and Integrin β3 (r=0.813,P =0.025). 18F-FDG uptake inversely correlated with the survival of tumor-bearing mice (r =0.500,P=0.017). Conclusions The uptake of 18F-FDGand 18F-FLT may reflect different biological characteritics of CRC. High 18F-FLT uptake in CRC on PET scan may predict high metastatic tendency.