Field avian infectious bronchitis virus (IBV) designated as JS/9 5/03, which was isolated from Jiangsu province of china, was cultivated in chicken emb ryo. It's single strain RNA was extracted from purified virus and worked as temp late of reverse transcription polymerase chain reaction (RT-PCR), a pair of pri mer designed according to megalign results of published IBV sequences in Genbank was used to amplify the neucleocapsid gene, the RT-PCR product was sequenced d irectly. Sequence analysis revealed that the sequence of JS/95/03 is most homolo gized with that of M41 strain.

Background Light-induced retinal damage models vary as many influence factors,herein the modeling method is difficult to copy.It is necessary to establish the grading standardization of retinal damage after retinal light exposure.Objective This study was to improve the modeling method and establish a grading standardization for light-induced retinal damage in rat.Methods Twenty-four SPF 8-10 week-old male SD rats were randomly divided into 4 groups and 6 eyes for each group.The rats were exposed to light intense of 5000 lx for 1,2,3 hours respectively in 3 groups,and other 6 rats served as the normal group.Full-field light exposure experiment was performed for each individual rat separately,and an annular illumination box was used tO ensure the experimental rat moving in a single direction and exposing the right eye in 5000 lx light surrounding during experimental duration.Ganzfeid electroretinogram(ERG)was recorded from the experimental rats at the fifth day after light exposure,and the animals were then sacrificed for histopathology observation to evaluate the retinal thickness change.All procedures which involved animals adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results After exposing to intensity light for 1,2,3 hours,the b-wave amplitudes of rod response,maximal mixed response,oscillatory potential in scotopic ERG as well as cone response,20 Hz flicker response of photopic ERG were significant declined as lapse of light exposure time(F=71.690,P=0.000；F=56.250,P=0.000；F=23.610,P=0.000：F=27.130,P=0.000；F=27.030,P=0.000)and lowed by 26.2％,52.5％,70.7％,24.4％,39.3％,58.1％respectively at the end of experiment.Meanwhile,the b-wave latencies of rod response,maximal mixed response in scotopic ERG as well as cone response of photopic ERG were evidently different among different groups (F=1.370,P=0.282；F：0.800,P=0.508；F=11.840,P=0.000；F=2.080,P=0.136).Light induced retinal damage located mainly at the temporal retina area.After intensity light exposure for 1,2,3 hours,the thickness of outer nuclear layer at the superior temporal retina attenuated by 11.3％,25.6％and 72.5％,respectively(P<0.05).A significant difference was seen in mean thickness of outer nuclear layer at superior temporal retina among different groups(F=410.27,P=0.000). Conclusion A standardized grading method for light-induced retinal damage is recommended.The continuous illumination in a intensity of 5000 Ix for 1,2,3 hours can induce the mild,moderate or severe retinal damage respectively at temporal retina.

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.
Amputees ; Arginase ; metabolism ; Arteriosclerosis Obliterans ; pathology ; Atherosclerosis ; pathology ; Autophagy ; Cell Polarity ; Chemokine CCL2 ; metabolism ; Foam Cells ; cytology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Macrophages ; cytology ; NF-kappa B ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phenotype ; STAT6 Transcription Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

OBJECTIVETo develop a colon-specific prodrug of Indomethacin microbially triggered, carry out in vitro/in vivo evaluation of drug release, and appraise its inhibitory effect on liver metastasis from colon cancer.

METHODSIndomethacin prodrugs were synthesized and characterized by FTIR and NMR, and dissolution test simulating gastrointestinal tract was employed to screen the colon-specific prodrug. Then, the pharmacokinetic profile of portal vein and peripheral blood in Sprague-Dawley rats was studied. Lastly, the inhibitory effect on liver metastasis from colon cancer in nude mice was observed.

RESULTSThe chemical structure characterized by FTIR and NMR demonstrated that six kinds of indomethacin-block-amylose with different drug loading (IDM-AM-1-6) were synthesized, among which IDM-AM-3 was degraded 1.3%, 9.3% and 95.3%, respectively, in simulated gastric fluid for 4 h, small intestine for 6 h, and colon for 36 h. The pharmacokinetic test of IDM-AM-3 showed that absorption was delayed significantly (P < 0.01), peak time [(11.35 + or - 2.45) h], elimination half-life [(16.74 + or - 4.04) h] and mean residence time [(22.27 + or - 0.52) h] were significantly prolonged (P < 0.01), as well as peak serum concentrations [(9.69 + or - 2.40) mg/L] and AUC(0-t) [(236.7 + or - 13.1) mg x L(-1) x h] were decreased markedly (P < 0.01) as compared with those of IDM regarding to portal vein. Additionally, its AUC(0-t) in peripheral blood was remarkably lower than that in Portal vein (P < 0.01). The tumor suppression observation showed that it could remarkably reduce the number of liver metastases in contrast to IDM (P < 0.05).

CONCLUSIONColon-specific IDM-AM-3 possesses advantage of sustained release in portal vein providing some experimental basis for colon-specific delivery system applied to sustained release in the portal vein.

Amylose ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Animals ; Colon ; metabolism ; Colonic Neoplasms ; pathology ; Delayed-Action Preparations ; Drug Delivery Systems ; HT29 Cells ; Humans ; Indomethacin ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Prodrugs ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley

This study aimed to assess the measurement uncertainty of a new method for determination of allura redin food by high performance liquid chromatography (HPLC).The uncertainty of mathematical model of allura red is based on Europe for Analytical Chemistry(EURACHEM) guidelines.The sources and components of uncertainty were calculated,including recovery,working solution,sample mass,final volume,response of standard solution,response of sample solution.The expanded uncertainty was 0.0024 (k=2).Uncertainty of working solution was the most significant factor contributing to the total uncertainty,accounting for 86.2％.The uncertainty of volume accounted for the minimum at 0.025％.The developed method is simple and accurate,which can be used for the determination of allura redin puffed samples.

OBJECTIVETo evaluate the antagonism effects of green tea (GT) against microcystin LR (MC-LR) induced hepatotoxicity and nephrotoxicity in mice.

METHODSAll 40 male mice were randomly divided into four groups. Mice in group III and IV were pretreated with green tea for free drink at doses of 2 g/L and 12 g/L prior to MC-LR intoxication, for consecutively 18 days. The toxin treatment mice were administered continually intraperitoneal injections of MC-LR at a dose of 10 microg x kg(-1) x d(-1) bw from day 6th till sacrifice, continually 13 days. Mice were sacrificed and immediately subjected to necropsy, and the body weight, relative organ weight, serum biochemical parameters, antioxidant enzyme levels (SOD and GSH), lipid peroxidation products (MDA) and histopathology were systematically evaluated.

RESULTSMC-LR exposure led to increase the oxidative stress and organ injury was significantly observed through biochemical parameters and microscopic evaluation. However, high dose of GT pretreatment caused a significant elevation in serum GSH and SOD levels, and a decrease of serum MDA level as compared with MC-LR control. The mean values of GSH and SOD activities were separately 467.29 mg/L and 139.22 U/ml in group IV. Subsequently, GT pretreatment obviously diminished the serum ALT, AST and Cr activities. Those pathological damages in liver and kidney, were to a certain extent, lessened in GT pretreatment mice in correlation with the biochemical parameters.

CONCLUSIONGT might elevate antioxidant defense system, clean up free radicals, lessen oxidative damages and protect liver and kidney against MC-LR induced toxicity.

Animals ; Antioxidants ; pharmacology ; Chemical and Drug Induced Liver Injury ; Free Radicals ; metabolism ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Liver Diseases ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Microcystins ; toxicity ; Oxidative Stress ; Tea

Animals ; Blood Glucose ; drug effects ; Crystallization ; Cyclohexanes ; chemistry ; pharmacology ; Female ; Hyperglycemia ; blood ; Hypoglycemic Agents ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Phenylalanine ; analogs & derivatives ; chemistry ; pharmacology ; X-Ray Diffraction

OBJECTIVETo establish and evaluate a rat model of type 2 diabetes mellitus (T2DM) associated with depression for further elaborating the disease.

METHODSTwenty-four Wistar rats were randomly divided into three groups: normal group (group N), T2DM group (group T) and T2DM with depression group (group T + D), with 8 rats in each group. The T2DM rat model was induced by high fat diet and low dose of Streptozotocin (STZ) injection, and in addition, the T2DM rats were made restraint stress for 21 days. After the model was established, the insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed. Then the rat depression level was analyzed by open field test, and the concentration of 5-hydroxytryptamine (5-HT) and dopamine (DA)was determined by ELISA to confirm the model identity.

RESULTSThe blood glucose level in group T and group T + D didn't return to the normal level at 180 minutes in the ITT and OGTT test; Compared with the group N, the max movement distance, retaining time in the central zone and the retaining frequency within 5 minutes in the group T + D decreased; 5-HT and DA level in the serum of rats in. group T + D was reduced.

CONCLUSIONA rat model of type 2 diabetes mellitus associated with depression has been successfully established by high fat diet and injection of low dose streptozotocin in combination with restraint stress for 21 days. This rat model is useful for further relevant studies.

Animals ; Depression ; complications ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; Diet, High-Fat ; Glucose Tolerance Test ; Rats ; Rats, Wistar ; Restraint, Physical ; Serotonin ; Streptozocin ; Stress, Psychological