Lipoxygenase ; metabolism ; Oxidation-Reduction ; Teratogens

Animals ; Cells, Cultured ; Excitatory Postsynaptic Potentials ; Glutamic Acid ; physiology ; Hippocampus ; cytology ; Neurons ; cytology ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the feasibility and diagnostic accuracy of CE-MRA with low dose contrast agent by comparison with DSA in diabetic patients with peripheral arterial diseases. Methods ( 1 )Study in vitro: test tubes containing Gd-DTPA of different concentrations were scanned, and the relationship between signal intensities and concentrations of GD-DTPA was analyzed. DSA and CE-MRA with selected concentrations of Gd-DTPA were performed on stenotic vascular models to estimate the proper low dose of GD-DTPA for clinical applications. (2) Clinical applications: 78 diabetic patients with peripheral arterial diseases were scanned from the abdomen and pelvis station to the calf-foot station in a 3 T MR system with standard bolus chase 3D CE-MRA sequence after injection of 13 ml GD-DTPA . The image quality,diagnostic rate of stenosis of arteries in calf and degree of venous contamination were evaluated with Fisher's exact test. DSA images of 220 vascular segments in 22 patients ( 10 segments per patient) were acquired as the gold standard and compared with CE-MRA by using Kappa test. Results The MR signal intensities were proportional to the concentrations of contrast agent in present study, and all stenotic segments of vascular model were displayed by CE-MRA with GD-DTPA at lower concentration of 1.5 mmol/L. As for MRA images of 78 diabetic patients with low dose Gd-DTPA, about 97.4% (76/78) showed diagnostic image quality for pelvic and thigh stations. But the MRA images of lower extremities were interfered by the venous contamination significantly (P ＜ 0.01 ). Compared with DSA for 22 patients, the diagnostic sensitivity, specificity and agreement coefficient (Kappa value) of MRA were 96. 0% ( 168/175), 73.3%(33/45), and 0.72 (P＜0.01), respectively. Conclusion Using 3.0 T MR scanner, high quality CE-MRA of lower limb arteries can be obtained for clinical applications with contrast agent dose as low as 13 ml,which has comparable diagnostic sensitivity and specificity with DSA. But the limitation of venous contamination in MRA image should be resolved in further studies.

Objective To establish a method to determine the contents of total flavonoids and shikimic acid in pine needles of Pinus massoniana Lamb.in Wudang Area.Methods Rutin was used as reference standard,and the content of total flavonoids in pine needles of Pinus massoniana Lamb. was determined by UV spectrometry at wavelength of 500 nm. The content of shikimic acid was determined by HPLC-DAD. The Fortis Xi C8 column (5 μm, 250 mm × 4.6 mm) was adopted with acetonitrile - 0.4% phosphoric acid solution (8:92, V/V) as mobile phase at the flow rate of 0.9 mL/min. The detection wavelength was 213 nm; the column temperature was 30 ℃; the injection volume was 20 μL. Results The linear range was 8.26-49.54 μg for rutin (r=0.999 4) with an average recovery of 99.2%, RSD=1.94%. The linear range was 10.26-61.56 μg for shikimic acid (r=0.999 4) with an average recovery of 99.5%,RSD=1.93%.The contents of total flavonoids in pine needles of Pinus massoniana Lamb.was 28.33 mg/g, and shikimic acid was 15.25 mg/g, respectively. Conclusion The method is simple, rapid, accurate and reproducible. It can be used for the content determination of total flavonoids and shikimic acid in pine needles of Pinus massoniana Lamb. in Wudang Area.

OBJECTIVEThe oxidation of benzo (a) pyrene mediated by 5-lipoxygenase (5-LOX) were investigated in HBE cells in order to provide further proof that lipoxygenase is the alternative pathway for the oxidation of xenobiotics.

METHODSEnzymic experiment: Soybean lipoxygenase (SLO), substrate (benzo[a] pyrene) and other component react in the enzymic system and the reaction product are detected by spectrophotometry. At the same time, in vitro detect of benzo (a) pyrene-DNA adducts with a UV spectrophotometer and HPLC. Cellular experiment: After HBE cells exposure to different poison (B[a]P 4, 8, 16, 32, 64, 128µmol/L, AA-861, naproxen or α- naphthoflavone 0.1, 1, 10 µmol/L) for 24 hours, the effect of benzo (a) -pyrene on cell survival rate were assessed by reductions of tetrazolium dye (MTT) and flow cytometry in cultured HBE cells, and the protein expressions of 5-lipoxygenase in the cells are tested by western-blot, and the DNA damages by the single cell gel electrophoresis. And then, the effect of the specific inhibitor of 5-lipoxygenase (AA-861) on 5-lipoxygenase protein expression and DNA damage in the cells are detected.

RESULTSSLO can catalyze the co-oxidation of benzo (a) pyrene to generate benzo (a) pyrene-7,8-epoxide in the presence of hydrogen peroxide. GTP can inhibit the reaction , the IC50 value is 0.46 mg/L, the model equation is Probit (P) = 0.8985+2.6824 Log (dose). SLO can catalyze the co-oxidation of benzo (a) pyrene to generate a new product, but fail to form DNA adducts in vitro. HBE cell viability decreased with the benzo (a) pyrene concentration increased , but AA-861 and naproxen can inhibit it. Flow cytometry and single cell gel electrophoresis experiments show, Benzo (a) pyrene can induce 5-lipoxygenase protein expression, but AA-861 cannot in HBE. Benzo (a) pyrene causes toxic action and DNA damage in HBE, which can significantly inhibit by AA-861, the difference is statistically significant (P < 0.05).

CONCLUSIONSThe co-oxidate of benzo (a) pyrene by 5-LOX turns into electrophiles that covalently bind to DNA and induce DNA damage, which can be significantly inhibited by AA-861.

Benzo(a)pyrene ; metabolism ; Cells, Cultured ; DNA Adducts ; metabolism ; DNA Damage ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Lipoxygenase ; pharmacology

ObjectiveTo probe the indication,treatment algorithm,anesthesia method,safety and efficacy ofextracorporealshockwavelithotripsy(ESWL)incombinationwithendoscopicretrograde cholangiopancreatography (ERCP) for pancreatic duct stones.MethodsThe patients with chronic pancreatitis and large pancreatic duct stones ( ＞ 5 mm diameter) and receiving ESWL and ERCP between March and July 2011 in Changhai Hospital were prospective studied.The third generation of extracorporeal shockwave lithotripsy was applied in ESWL,and the localization of stone was determined by X-ray.No more than 5000 shocks were given per session,and ESWL was performed continuously till the calculi were fragmented,and then was cleared by ERCP.ResultsA total of 100 patients underwent ESWL during the 5 months,among whom 84 patients received ERCP treatment and 41 cases failed to deep cannulation (41/84,48.8％ ).Multiple stones were seen in 83 patients.Ninety five patients had radio-opaque stones,two patients had radiolucent calculi,while three patients had both radio-opaque and radiolucent stones.Seventy five percent,14％ and 11％ stones were located in pancreatic head,pancreatic head and body,pancreatic body and tail,respectively.A total of 175 ESWL procedures were performed,43 patients needed 2 or more sessions for successful fragmentation.Anesthesia method was mainly intravenous sedation,accounting for 96％ (168/175).ERCP was successful in 96 patients after ESWL,only 4 patients failed after ESWL. Forty one cases which failed ERCP procedures before ESWL underwent ERCP,and 37 patients (90.2％) achieved successful cannulation.Successful fragmentation ratewas 100％.Complete clearance was achieved in 78 patients,and complication rate of post-ERCP pancreatitis,fever was 1.71％ (n =3 ),0.57％ (n =1 ),and the overall complication rate was 2.28％.Conclusions ESWL is an effective,safe and necessary modality for fragmentation of large PD stones in the management of minimal invasive treatment of chronic pancreatitis.

Interleukin-2 (IL-2) was initially isolated as a T cell growth factor and had been shown to direct the expansion and differentiation of several hematopoietic cell types. Clinical studies using IL-2 in the treatment of AIDS have been encouraging, due to its critical role as a proliferative signal for activated T-lymphocytes. IL-2 has also undergone trials in the treatment of several types of cancer, based on its stimulation of cytotoxic, antitumor cells. Today, human IL-2 is produced completely by genetically engineered method, and it has been proved that genetically engineered recombinant human IL-2 has almost the same function and clinical effect as wild IL-2. In the former study, recombinant human IL-2 usually comes from E. coli, in this paper the mutant IL-2 was successfully expressed and purified in Pichia pastoris for the first time. As a eukaryote, Pichia pastoris has many of the advantages of higher eukaryotic expression systems such as protein processing, protein folding, and posttranslational modification, while being as easy to manipulate as E. coli or Saccharomyces cerevisiae. It is faster, easier, and less expensive to use than other eukaryotic expression systems such as baculovirus or mammalian tissue culture, and generally gives higher expression level. Expression conditions of human mutant interleukin-2(the codon for cysteine-125 of human IL-2 with alanine; the codon for leucine-18 with methionine; the codon for leucine-19 with serine) in the recombinant Pichia pastoris strain were optimized via test of some factors such as the rate of aeration, the inductive duration, the initial pH and the concentration of methanol. The results from tests showed that the most important parameter for efficient expression of interleukin-2 in recombinant Pichia pastoris strain is adequate aeration during methanol induction, and the optimum inductive condition for interleukin-2 expression was: more than 80% aeration, 2 days for induction, the initial pH of 6.0, the final methanol concentration of 1.0%. With this condition, the expressed IL-2 was secreted into fermentation broth and reached a yield of 30%, approximately 200 mg/L. Expressed interleutin-2 (MvIL-2) was isolated and purified by centrifugation, millipore filtration to concentration, Econo-PacS strongly acidic cation exchanger cartridge and molecular sieve chromatography and the yield of MvIL-2 was 27%. MvIL-2 was purified to electrophoretic purity by SDS-PAGE and only one peak being loaded on HPLC. Purified MvIL-2 protein had stimulating activity similar to the wild type of IL-2 as assayed by IL-2-dependent CTLL-2 cells. However, the stability of MvIL-2 was superior than that of IL-2 at different temperatures. The activity of obtained MvIL-2 was 4 - 5 times of the wild type of IL-2, So MvIL-2 had an advantage over wild type of rhIL-2 in storage stability and activity.
Fermentation ; Humans ; Interleukin-2 ; biosynthesis ; genetics ; Mutant Proteins ; biosynthesis ; Mutation ; Pichia ; genetics ; metabolism ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

OBJECTIVETo study the death mode of human hepatocellular carcinoma Hep-3B cells and human lung adenocarcinoma A549 cells induced by bluetongue virus strain HbC3 (BTV-HbC3) and the mechanism of its action.

METHODSBTV-HbC3 was used to infect the tumor cells, and the cytopathic effects (CPE) was observed. TUNEL staining was used to detect the apoptosis of tumor cells induced by BTV-HbC3. The changes of endoplasmic reticulum and nuclei treated with BTV-HbC3 were further examined by laser scanning confocal microscopy. The activities of caspase-3/7, caspase-8 and caspase-9 were determined by fluorescence analysis.

RESULTSHep-3B cells were sensitive to BTV-HbC3. Lots of early apoptotic cells were found by TUNEL staining. The laser scanning confocal microscopic examination showed characteristics of apoptosis, such as pyknotic nuclei, margination of nuclear chromatin and vacuolization of endoplasmic reticulumin in Hep-3B cells exposed to BTV-HbC3. The activity of caspase-3/7 was increased, but the activity changes of caspase-8 and caspase-9 were not found. A549 cells were sensitive to BTV-HbC3 too. But no apoptotic cells were observed by TUNEL staining. The results of laser scanning confocal microscopy showed marked vacuolization of endoplasmic reticulum, but chromatin margination was not found after A549 cells was exposed to BTV-HbC3. The activity of caspase-3/7 and caspase-9 was increased, but the activity of caspase-8 was not changed.

CONCLUSIONBTV-HbC3 induces apoptosis of Hep-3B tumor cells mainly through endoplasmic reticulum signal transduction pathway, and the features of cell death in A549 cells could be described as paraptosis.

Adenocarcinoma ; metabolism ; pathology ; virology ; Apoptosis ; Bluetongue virus ; pathogenicity ; physiology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; virology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Nucleus ; pathology ; Endoplasmic Reticulum ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; virology ; Lung Neoplasms ; metabolism ; pathology ; virology ; Oncolytic Viruses ; pathogenicity ; physiology ; Signal Transduction

This study was intended to look for anti-HIV chemical constituents of aerial parts of Caragana rosea Turcz. Column chromatographic technique was used for the isolation and purification of constituents of Caragana rosea under the guide of anti-HIV assay. The structures were established on the basis of physical and chemical properties and spectroscopic data. Five compounds were obtained from the EtOAc fraction of aerial parts of Caragana rosea and identified as myricetin (1), mearnsetin (2), p-hydroxy cinnamic acid (3), cararosinol A (4) and cararosinol B (5). At the same time, one possible transformation route between cararosinol B and kobophenol A, another resveratrol tetramer isolated from this plant previously, was proposed. Compounds 4, 5 are new resveratrol tetramers, compounds 1 -3 were isolated from this plant for the first time. All compounds showed no activities in an in vitro assay against HIV-1.
Anti-HIV Agents ; chemistry ; isolation & purification ; pharmacology ; Benzofurans ; chemistry ; isolation & purification ; pharmacology ; Caragana ; chemistry ; Coumaric Acids ; chemistry ; isolation & purification ; pharmacology ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; HIV-1 ; drug effects ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Propionates ; Stilbenes ; chemistry ; isolation & purification ; pharmacology

Inguinal hernia is the most prevalent type of abdominal wall hernia. Indirect inguinal hernia is twice as common as direct inguinal hernia. Computed tomography and magnetic resonance imaging can be used to evaluate inguinal hernia, but these modalities are greatly limited by their cost and availability. Ultrasonography has emerged as the most convenient imaging tool for diagnosing inguinal hernia due to its advantages, such as portability and absence of radiation. The present pictorial review presents an overview on the use of ultrasonography in the evaluation of inguinal hernia with a particular emphasis on the regional anatomy, relevant scanning tips, identification of subtypes, postoperative follow-up, and diagnosis of pathologies mimicking inguinal hernia.