Adenine Research Articles

Changes of NADPH oxidase expression in retina of rd mice

Background Studies showed that activation of microglia-derived nicotinamide adenine dinucleotide phosphate (NADPH) oxidase plays a key role in the neurodegenerative diseases and neural cell death in central nervous system.The effect of NADPH on cone degeneration have been determined in rd rats,its role in rod degeneration is relatively less studied.Objective This study was to study the expression of NADPH oxidase in the retinal degenerative process in rd mice and further explore its role in the photoreceptor degeneration.Methods rd Mice at postnatal day 8 (P8),P10,P12,P14,P16 and P18 were collected.The mice were sacrificed,and retinal sections,RNAs and proteins were prepared in above-mentioned time points.The expressions of the gp91 phox,a major subunit of NADPH oxidase,in transcript level and protein level in the retinas were semi-quantitatively detected by real-time PCR and Western blot respectively.Expression of gp91phox was localized in the rd retinas as ageing by immunohistochemstry,and the co-expression of gp91phox with CD11b,a specific marker of microglial cells,was assayed by immunofluorescent double labeling.The C57BL/6N mice were served as controls.The use and care of the animals complied with the Guideline of ARVO.Results Real-time PCR showed that gp91phox mRNA was not expressed in the retinas of C57BL/6N mice.Gp91phox mRNA was found to have less expressed in retinas of P8 rd mice.With aging,the expression level of gp91phox mRNA (gp91phox mRNA/β-actin) in rd mouse retinas was gradually increased with the highest level in P14 mice(1.136±0.370).A significant difference was seen in the gp91 phox mRNA expression among various groups of mice (F=17.81,P =0.00),and gp91phox mRNA expression was significantly elevated in P10,P12,P14,P16 and P18 rd mice compared with P8 rd mice(all at P＜0.05).The expression level of gp91phox protein (A value) in the retinas presented with a similar trend in the rd mice,with a significant difference among the various ages of rd mice and C57BL/6N mice (F =354.00,P＜0.01).The expression level of gp91 phox protein was increased in the rd mice in comparison with the C57BL/6N mice (all at P＜0.05).Immunochemistry revealed that the positive response cells for gp91phox increased in the inner layers of retinas in P10 rd mice and peaked in P14 mice.Immunofluorescent double labeling exhibited that gp91phox were seen to present a co-expression with CD11b,showing an orange fluorescence.Conclusions Expression of NADPH oxidase in the rctinas in the rd mice up-regulates and is parallels to the microglial activation and photoreceptor degeneration,suggesting that NADPH oxidase plays a role in the retinal dystrophy associated with microglial activation. Key words: Nicotinamide adenine dinucleotide phosphate oxidase; Retinal degeneration; Microglia; rd Mouse/Nspontaneous hereditary retinal degeneration

A thixotropic supramolecular hydrogel of adenine and riboflavin-5′-phosphate sodium salt showing enhanced fluorescence properties

An equimolar mixture of riboflavin-5′-phosphate sodium salt (RP) and adenine (AD) dissolved in a phosphate buffer (pH 4.0, 1.0% w/v) produces a red coloured transparent thixotropic hydrogel at 30 °C. The gelation of the RPAD system occurs in the pH range of 2–5. FTIR spectra and WAXS patterns indicate self-assembly via H-bonding between the >C=O group of RP and the amino/imino group of AD followed by supramolecular organization through a π-stacking process producing a fibrillar network structure. FESEM images clearly indicate that the nanofibres are produced from the intertwining of helical fibrils. The dynamic frequency sweep experiment of the supramolecular gel at a constant strain of 1% exhibits a wide linear viscoelastic region and a considerably higher G′ value (460 Pa) than that of G′′ (21 Pa) confirming the gel nature of the RPAD system. The hydrogel shows high stiffness (G′/G′′ = 3.3), a high yield stress (σ*) (79.5 Pa) and a moderate critical strain (γ = 17.5%). Time sweep experiments at both low (0.1%) and high strain (100%) indicate the thixotropic property of the gel. The RPAD hydrogel shows non-Newtonian viscosity in the shear rate region (0.1–158 s(−1)) and after that there is a sudden fall of viscosity. The gel melting point obtained by the falling ball method is 6° higher than that obtained by the DSC method probably due to the presence of the thixotropic property of the gel. The UV-vis spectra indicate a red shift of the π–π* transition band of RP in the RPAD xerogel. On excitation of the RPAD hydrogel at 373 nm it shows twelve times enhancement of emission intensity with a 7 nm red shift of the emission peak. This has been attributed to the enhancement of lifetime from 2.2 ns in RP to 3.4 ns in the RPAD hydrogel. With increase of temperature, the fluorescence intensity of the RPAD hydrogel at first increases till 40 °C, then decreases up to 55 °C and it again increases after 60 °C.

Roles of SIRT1 in tumorigenesis and tumor progression

Silent information regulator 1 (SIRT1) is a kind of histone deacetylase which dependents on nicotinamide adenine dinucleotide.It is involved in the occurrence and development of tumors.Data show that SIRT1 is highly expressed in many tumors,such as gastric cancer,breast cancer,prostate cancer and so on.On the other hand,the expression of SIRT1 is lower in many other types of tumors,including glioblastomas,bladder cancer,ovarian cancer and so on.Therefore,the role of SIRT1 in tumor formation process is still uncertain and controversial. Key words: Neoplasms; Silent information regulator 1

Neuroprotective effect of lycium barbarum polysaccharides after cavernous nerve crush injury in a rat model

Objective To investigate the feasibility of erectile function recovery by repairing crushed cavernous nerves (CN) with lyceum barbarum polysaccharides (LBP).Methods Twenty-one male Sprague-Dawley rats were randomly divided into 3 groups:sham group,LBP group,and control group.Erectile function was assessed by cavernous nerve electrostimulation at 3rd month after operation and nerve regeneration was assessed by toluidine blue staining of CN and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue.Results The peak ICP/MAP ratio in LBP group (0.44 ± 0.05) was significantly higher than in control group (0.26 ± 0.03,P ＜ 0.05),but lower than in sham group (0.68-± 0.08,P ＜ 0.05).The LBP group had more myelinated axons of CNs (106.8 ± 17.9) than the control group (64.9 ± 13.9,P ＜ 0.05),but fewer than the sham group (180.9 ± 15.3,P ＜ 0.05).The LBP group had more NOS diaphorase positive nerve (164.2 ± 12.2) than the control group (85.6 ±14.7,P＜0.05),but fewer than the sham group (263.3 ± 15.0,P＜0.05).Conclusion The application of LBP after CN crush injury promotes nerve regeneration and erectile function recovery. Key words: Lyceum barbarum polysaccharide; Erectile dysfunction; Cavernous nerve

Glutathione-enriched baker's yeast: production, bioaccessibility and intestinal transport assays

A glutathione (GSH) yeast-based biomass (Saccharomyces cerevisiae) was used to investigate GSH stability, solubilization during gastrointestinal digestion and GSH intestinal transport. A postgrowing procedure was applied to improve intracellular GSH yeast content. The presence of adenine (ADE) in the biotransformation solution (CYS-GLY-GLU mixture) and alternatively, a glucose shot after 4-h incubation, allowed to obtain cells containing about GSH 1.6-1.7% dcw (dry cell weight) (control 0.5%). Yeast samples were subjected to in vitro gastrointestinal digestion and absorption assays employing Caco-2 and HT29-MTX cell lines in different proportions (100/0, 70/30 and 50/50). Trials were also performed to verify intestinal cell viability. At least 87% of ingested GSH is available in reduced form for intestinal absorption. In vitro GSH transport assays indicated that GSH is poorly absorbed (<20%). Nevertheless, studies in response to oxidative stress induced by H2 O2 demonstrated a protective role of the GSH-enriched biomass towards intestinal cell viability. An enriched GSH yeast-based biomass has been obtained using a postgrowing procedure. Although GSH present in enriched yeasts is poorly absorbed by intestinal cells, this biomass showed an intestinal local protective effect, improving cells viability when a simulated oxidative stress was applied.

Graphene Based Microsensors for the Assay of Adenine, Guanine and Epinephrine

not Available.

Effect of celastrol postconditioning on focal cerebral ischemia-reperfusion injury in rats

Objective To evaluate the effect of celastrol postconditioning on focal cerebral ischemiareperfusion (I/R) injury in rats.Methods Sixty-four Sprague-Dawle rats (32 males,32 females),weighing 250300 g,were randomized into 4 groups using a random number table (n =16 each):sham operation group (group S) ; celastrol control group (group S + C) ; focal cerebral I/R group (group I/R) ; celastrol postconditioning group (group I/R + C).Focal cerebral I/R were produced by middle cerebral artery occlusion (MCAO).Dimethyl sulfoxide (DMSO) 0.3 ml/kg was injected intraperitoneally after shame operation in group S.Celastrol 3 mg/kg was injected intraperitoneally after shame operation in group S + C.DMSO 0.3 ml/kg was injected intraperitoneally at 5 min of reperfusion in group I/R.Celastrol 3 mg/kg was injected intraperitoneally at 5 min of reperfusion in group I/R + C.The neurologic deficit was scored at 5 min before reperfusion and 24 h of reperfusion.The infarct size was detected by TTC staining,and then the percentage of infarct size was calculated.The pathological changes in CA1 region of ischemic hippocampus were detected by HE staining.The activity of nicotinamide adenine dinucleotide phosphate oxidase (NOX),content of reactive oxygen species (ROS) and expression of NOX1 and NOX2 mRNA (by RT-PCR) in ischemic brain tissues were detected.Results Compared with S and S + C groups,the neurologic deficit scores,infarct size,percentage of infarct size,NOX activity and ROS content were significantly increased,and the expression of NOX1 mRNA and NOX2 mRNA was up-regulated in I/R and I/R + C groups (P ＜ 0.01).Compared with group I/R,the neurologic deficit scores,infarct size,percentage of infarct size,NOX activity and ROS content were significantly decreased,and the expression of NOX1 mRNA and NOX2 mRNA was down-regulated in group I/R+ C (P ＜ 0.01).There was no significant difference in the parameters mentioned above between group S and group S + C (P ＞ 0.05).The pathological changes in CAl region of ischemic hippocampus were significantly attenuated in group I/R + C (P ＜ 0.01).Conclusion Postconditioning with celastrol can auenuate focal cerebral [/R injury in rats and inhibiton of oxidative stress response in brain tissues may be involved in the mechanism. Key words: Tripterygium; Brain; Reperfusion injury; NADPH oxidase; Reactive oxygen species; Ischemic postconditioning

Intraoperative rapid diagnosis of Hirschsprung's disease

Objective To evaluate the superiorities and disadvantages of reduced nicotinamide adenine dinucleotide dehydrogenase (NADPH-d),acetylcholinesterase (AchE) enzyme histochemistry and hematoxylin and eosin (HE) staining in the intraoperative rapid diagnosis of Hirschprung' s disease (HD),increase the intraoperative diagnostic accuracy and decrease the risks of reoperation.Methods Eighteen children with HD underwent intraoperative full-thickness intestinal biopsies,including 11 boys,and 7 girls,with average age of (8.6 ± 3) months.Frozen sections at three levels (narrow segments,transitional region and dilated segments) were taken,and the extent of aganglionosis was examined by using NADPH-d staining,traditional rapid HE and AchE staining respectively.TUJ-1,a marker of enteric neural cells,was used for identification of NADPH-d positive myenteric plexus and ganglion cells by immunofluorescence staining.Results The positve rate of NADPH-d staining,HE staining and AchE staining for HD was 94.4％ (17/18),83.3％ (15/18),and 88.9％ (16/18),respectively (P ＞ 0.05).Time comsuming in NADPH-d,HE and AchE staining was (7.0 ± 1.7),(23.2 ± 2.9) and (30.9 ± 2.2) min,respectively(P ＜ 0.01).Conclusion The combination of NADPH-d and HE staining,with high idiosyncrasy and sensibility,simple and convenient,can be used as a routine intraoperative diagnosis method to identify HD and detect cutting range of colon.AchE staining was the best technique to demonstrate hypertrophic nerve trunks in the mucosa and submucosa and was well suited to suction rectal biopsy. Key words: Hirschprung's disease ; Intraoperative diagnosis; Pathology ; Colorectal surgery

Expression of nicotinamide adenine dinucleotide phosphate oxidase 5 in carotid atherosclerotic plaques of a swine model

Objective To assess the association of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) 5 expression with plaque vulnerability in a carotid atherosclerotic swine model.Methods Carotid atherosclerosis models were created in 5 Chinese miniswines using the combination of partial ligation and high cholesterol diet for 3 months.Animals were sacrificed to obtain bilateral carotid arteries and rete mirahile for histological analvsis.The relationship between morphological features of carotid plaques and NOX5 expression was also evaluated.Results Atherosclerotic plaques were developed in all ten carotid arteries.Typical features of unstable plaques were also found.Distal embolisms were detected in all 10 rete mirabilis.NOX5 was expressed in both the early atherosclerotic lesions and advanced plaques.The NOX5 expression was increased in the advanced carotid plaques with fibrous cap rupture as compared with that in those without fibrous cap rupture (P ＜ 0.05).Conclusion Carotid atherosclerosis model with the features of vulnerable plaque and distal embolism can be created by the combination of partial ligation and high cholesterol diet in Chinese miniswiues.Increased expression of NOX5 may be associated with vulnerable carotid plaques. Key words: Atherosclerosis; Carotid artery; Model, animal; Swine; Nicotinamide adenine dinucleotide phosphate oxidase

Plant regeneration via somatic embryogenesis and organogenesis in Verbascum speciosum Schard

Plant regeneration was achieved in Verbascum speciosum Schard. via organogenesis and somatic embryogenesis by culture of mature embryo explants. Two types of calli, embryogenic and non-embryogenic, were induced from mature embryo explants on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyl adenine (BA) and α-naphthalene acetic acid (NAA). In order to further proliferate the somatic embryoids, the yellow and friable embryogenic calli were transferred on MS medium containing 0.5 mg-1 charchol and 0.1 or 1 mg-1 2,4-dichlorophenoxy acetic acid (2,4-D) or into MS medium containing 60 g-1 sucrose, 50 mgl-1 casein hydrolysate (CH), 0.5 mg-1 kinetin (Kin), 5 mg-1 2,4-D and 0.5 mg-1 charchol. Among the 3 tested media, MS medium containing 0.1 mg-1 2,4-D and 0.5 mg-1 charchol was more effective for proliferation of embryonic calli. Somatic embryos were transferred to hormone free MS medium for maturation and shoot regeneration. In addition, shoots and roots regenerated from non-embryogenic calli in hormone free MS medium or containing NAA and BA. Shoot buds were obtained from non-embryogenic calli and they were transferred to MS medium supplemented with 1 mg-1 BA or Kin for further growth and multiplication. Regenerated plants then were potted and maintained in the greenhouse.

In vitro propagation and phytochemical assessment of Berberis chitria : An important medicinal shrub of Kumaun Himalaya, India

Berberis chitria is an important Himalayan shrub known for its diverse medicinal uses. A reproducible in vitro micropropagation protocol for B. chitria has been developed. Seeds were initially inoculated on water agar medium; contamination-free green seeds were transferred to Woody Plant Medium (WPM) with 78.89% seed germination. Initially, maximum multiple shoots (8.30±1.0) per explant were observed in WPM supplemented with 6-benzyl adenine (BA)+α-Naphthalene acetic acid (NAA) (8.88+1.34 µM). The shoot multiplication rate was increased up to 22.89±2.51 shoots per explant on successive subculture but shoot necrosis and concomitant decrease in shoot height was the big problem. Incorporation of casein hydrolysate (CH; 500 mg/L) and gibberellic acid (1.44 µM) in shoot multiplication medium not only facilitated shoot height, but the number of shoots per explant also increased without any necrosis. Microshoots were exposed to auxins in two different ways. The highest rooting percentage (100%) was observed after 8 weeks on WPM supplemented with indol butyric acid (IBA; 100 µM) for 7 days with 5.92±0.36 roots per microshoot. Well-rooted plants were transferred to thermacol pots containing nonsterile, sieved soil and farmyard manure and hardened successfully. This study also provides evidences that the methanolic extract of in vitro raised plants have strong antioxidant activities and higher phenol content than naturally grown plants. Key words: Berberis chitria, 6-benzyl adenine (BA), gibberellic acid (GA3),micropropagation, antioxidant activity.

Effects of quercetin on gene and protein expression of phagocytic nicotinamide adenine dinucleotide phosphate oxidase and nitric oxide synthase after myocardial ischemia and reperfusion in rabbits

Objective Investigate the effects of quercetin on phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX2),endothelial nitricoxide synthase (eNOS) and inducible nitricoxide synthase (iNOS) after myocardial ischemia-reperfusion injury (MIRI) in rabbit.Methods New Zealand rabbits were subjected to 30 minutes of myocardial ischemia followed by 12 hours of reperfusion.They then were randomly assigned to four experimental groups:control,L/R,quercetin (Que),I/R + Que.Gene and protein expression of NOX2,eNOS and iNOS were compared.Results mRNA and protein expression of NOX2 in I/R were 0.29 ± 0.11,0.29 ±0.11,mRNA and protein expression of iNOS in I/R were 0.29 ± 0.11,0.29 ± 0.11.Both in Real-time polymerase chain reaction (Real-time PCR) and Westem blotting studies,myocardial ischemia reperfusion-induced NOX2 and iNOS expression were enhanced (P ＜0.01),but eNOS mRNA and protein in I/R hearts were not significantly different from that in control (P＜0.01).After administration of quercetin,mRNA and protein expression of NOX2 in Que were 0.40 ± 0.08,0.56 ± 0.28,mRNA and protein expression of eNOS in Que were 0.22 ± 0.07,0.95 ±0.23,mRNA and protein expression of iNOS in Que were 0.02 ±0.01,0.87 ±0.31 ; mRNA and protein expression of NOX2 in I/R + Que were 1.03 ±0.31,2.05 ±0.49,mRNA and protein expression of eNOS in I/R + Que were 0.32 ± 0.09,1.12 ± 0.36,mRNA and protein expression of iNOS in I/R + Que were 0.16 ±0.05,3.17 ± 1.29.Administration of quercetin reduced NOX2,eNOS and iNOS mRNA and protein expression both in control and I/R heart (P ＜ 0.01).Conclusion Gene and protein expression of NOX2 and iNOS were increased after MIRI.Quercetin not only inhibited myocardial ischemia reperfusioninduced NOX2 and iNOS mRNA and protein expression but also eNOS mRNA and protein expression. Key words: Ischemia-reperfusion injury; Quercetin; Phagocytic nicotinamide adenine dinucleotide phosphate oxidase; Endothelial nitricoxide synthase; Inducible nitricoxide synthase

In vitro plant regeneration from epicotyl explant of Withania somnifera (L.) Dunal

Studies were carried out to investigate the regeneration and rapid multiplication of Withania somnifera (L.) in vitro. Direct and indirect regeneration protocols for multiple shoots development from epicotyl explants of 50 to 60 days old seedlings were established. The shoots were initiated directly from epicotyl explant on 6-benzyl amino purine (BAP: 2.0 mg/L) along with indole-3-acetic acid (IAA: 0.2 mg/L), and the maximum of 15.5 ± 0.90 shoots/explant were achieved by subsequent subcultures at 4 weeks interval in the same medium. Calli (98.3%) were produced from epicotyl explant on 2,4-dichlorophenoxy acetic acid (2,4-D: 2.0 mg/L) along with kinetin (Kn: 0.6 mg/L), and shoots were initiated from calli on BAP (1.0 mg/L) along with adenine sulphate (AdS: 20.0 mg/L). Proliferation of shoots was achieved by subsequent subcultures at 4 weeks interval in the same medium. The maximum value of 25.3 ± 1.81 shoots/explant was achieved in the second subculture of indirect regeneration. Murashige and Skoog (MS) medium along with gibberellic acid (GA3) at 1.0 mg/L produced maximum 73.3 and 95.5% of shoot elongation in direct and indirect regenerated shoots, respectively. On the other hand, MS medium with indole-3-butyric acid (IBA) at 0.8 mg/L induced maximum 86.7% and 90.0% of rooting from elongated shoots of direct and indirect regeneration, respectively. The rooted plants were transferred to small cups filled with sterilized mixture containing soil, sand and vermiculite (1:2:1, v/v/v) for hardening. About 90% of plants survived in the hardening process, and then the plants were established successfully in the experimental field. This protocol yielded a higher number of shoots within a short period. Consequently, the protocol developed in this study offers a simple and improved in vitro method to regenerate W. somnifera. Key words: Withania somnifera, epicotyls, tissue culture, shoot elongation, root induction.

TARGETING OF NAADP-MEDIATED CALCIUM SIGNALING AFFECTS VEGF-INDUCED ANGIOGENESIS

Vascular endothelial growth factor (VEGF) and its transmembrane receptors VEGFR1 and VEGFR2 play a key role in controlling both physiological and pathological angiogenesis, including vascularization of solid tumours. We have identified a novel and crucial signalling mechanism through which activation of VEGFR2 in human endothelial cells (HUVEC) selectively triggers the intracellular release of calcium from acidic compartments, operated by the second messenger NAADP (nicotinic acid adenine dinucleotide phosphate). Live imaging of calcium fluxes in cells treated with VEGF in the presence of specific inhibitors have shown that 1) VEGF-activated calcium stores are different from IP3 and ryanodine sensitive compartments and are of acidic nature, which strongly indicates the involvement of NAADP signalling 2) NAADP inhibition by its specific antagonist Ned-19 abolishes VEGF-induced calcium response. This inhibition is accompanied by impaired phosphorylation of downstream targets ERK1/2, Akt, eNOS, JNK (but not p38) and results in significant reduction of cell proliferation, migration and capillary-like tube formation in vitro. Interestingly, when the angiogenic response to VEGF was assayed in vivo utilizing Matrigel plugs subcutaneously implanted in mice, Ned-19 was found to dramatically inhibit VEGF-induced angiogenesis. Altogether our data showing that NAADP plays a key role in the control of VEGF-induced angiogenesis could potentially contribute to identify new targets for antiangiogenic therapeutic strategies, a goal to which much scientific effort has long been devoted but still awaiting ultimate success.

Sirtuin 6 localization at cortical brain level of young diabetic mice

The metabolic syndrome, characterized by visceral obesity, dyslipidaemia, hyperglycaemia and hypertension, has become one of the major public-health challenges worldwide and it is strictly associated with the development of type II diabetes and neurodegenerative diseases (Alberti et al. 2005; Panza et al. 2010). Increased metabolic flux to the brain during overnutrition can orchestrate stress response, blood-brain barrier alteration, microglial cells activation and neuroinflammation (Nerurkar et al., 2011). The protein sirtuin family is a class of nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase that act on a variety of targets and so play a key role in central physiological regulation (Sebastian et al., 2012; Wang et al., 2012). To assess the physiopathological significance of sirtuin6 (SIRT6) at brain cortical level, we analysed its specific expression and subcellular localization in young db/db mice, animal model of type II diabetes mellitus, and respective control lean mice. In particular, we analysed the cytoarchitecture of the brain cortex, evaluated SIRT6 expression and its localization by immunohistochemistry comparing young db/db mice to lean control mice, distinguishing among the six cortical layers and between motor and somatosensory cortex. We observed that SIRT6 is mainly localized in the nucleus of both lean and db/db mice. Diabetic mice showed few SIRT6 positive cells respect to lean control mice in all cortical layers without significant differences between motor and somatosensory cortex. No morphological alteration have been find. In conclusion, our findings contribute to further understand SIRT6 protein expression in the early steps of type II diabetes mellitus and suggest its implication in the pathogenic processes of diabetes mellitus and diabetes–induced neurodegeneration.

Determination of D816V mutation in the c-kIt gene in the Slovenian patients with acute myeloid leukemia and systemic mastocytosis

Background: D816V mutation in the C-KIT gene is present in more than 90 % of patients with systemic mastocytosis (SM) and 2–7 % of patients with acute myeloid leukemia (AML). D816V mutation is caused by the substitution of adenine with thymine at 2447 nucleotide sequence in the C-KIT gene. This nucleotide substitution causes the replacment of aspartate acid by valine at codon 816 of the KIT protein. KIT protein with D816V mutation acts as constitutively active tyrosine kinase that promotes cell proliferation and inhibits apoptosis. The purpose of our study was to determine the incidence of D816V mutation in the C-KIT gene in Slovenian patients with AML and in patients with suspected systemic mastocytosis. Patients and methods: In the retrospective study, 71 patients with AML and 25 patients with suspected systemic mastocytosis were included. D816V mutation in the C-KIT gene was determined by polymerase chain reaction (PCR) and the resulting PCR products were analyzed by agarose gel electrophoresis. Results: D816V mutation in KIT protein was determined in 7 % of patients with AML and in 32 % patients with suspected systemic mastocytosis. Conclusions: Identification of D816V mutation in the C-KIT gene must always be performed in patients with suspected systemic mastocytosis. The determination of this mutation contributes to the diagnosis and treatment selection. The finding of D816V mutation in the C-KIT gene in patients with AML and concomitant genetic modifications RUNX-RUNX1T1 (typical translocation t(8; 21) (q22, q22)) or CBFB-MYH11, which is the result of inversion on chromosome 16–(inv (16) (p13, q22)), however, indicates a faster, more aggressive course of the disease and predicts a worse outcome. The finding of the mutation in other patients with AML may indicate the presence of concomitant AML and SM, which was not found in our patients.

Experimental Investigation of the Inhibiting Action of Adenine on the Corrosion of Mild Steel in Acidic Environments

Corrosion inhibition of mild steel by adenine (AD) in 0.1 M solutions of HCl and H2SO4 was investigated using weight loss, potentiodynamic polarization and electrochemical impedance spectroscopy techniques. The study was carried out at 30 °C and 60 °C respectively. The effect of iodide ions on inhibition efficiency was also assessed. Results showed that AD had a moderate inhibitive effect on the acid corrosion of mild in the order HCl > H2SO4 at 30 °C and H2SO4 > HCl at 60 °C. The presence of iodide ions in the acid media increased the efficiency of AD. From potentiodynamic polarization studies, it was observed that AD alone and the mixture of AD+KI surpressed both the anodic and cathodic partial reactions. The corrosion processes were inhibited by adsorption of AD onto the mild steel surface in a concentration and temperature-dependent manner. Langmuir isotherm was used to explain the adsorptive characteristics of AD.

Sirtuin 1 in skin diseases

Sirtuin 1 （SIRT1）, a member of the conserved sirtuin family, is a nicotinamide adenine dinucleotide （NAD）-dependent histone and nonhistone deacetylase, participating in the regulation of longevity, energy metabolism, stress, inflammation, tumor, and so on. In recent years, some scholars have found that SIRT1 can protect subepithelial collagen and prevent ultraviolet irradiation-induced photoaging, and also play an important anti-inflammatory effect via down-regulating cytokines such as STAT3 and COX2. The relationship between SIRT1 and skin tumors is complex, and SIRT1 may act as either an oncogene or a tumor- suppressor gene. SIRT1 activators and inhibitors may play an important role in the treatment and prevention of skin diseases in future. Key words: Sirtuins; Light; Cell aging; Skin neoplasms

Efficient propagation and rooting of three citrus rootstocks using different plant growth regulators

The influence of various basal medium and plant growth regulators on the efficient micropropagation of nodal explants from mature trees of alemow, sour orange, and ‘Cleopatra’ mandarin citrus rootstocks was studied. All three citrus rootstock shoot cultures showed a preference for high-salt media, like Murashige and Skoog or Driver and Kuniyuki Walnut medium. Several combinations of N 6-benzyladenine (BA) and adenine (AD), kinetin (KIN) or gibberellic acid (GA) were tested to optimize the shoot proliferation phase. BA/GA combinations improved the proliferation of all the rootstocks studied, especially alemow. The addition of BA and AD to the culture medium improved shoot proliferation in sour orange and ‘Cleopatra’ mandarin in the same way as BA and GA. The addition of different combinations of BA/KIN did not result in further improvement of any of the studied variables. The transfer of in vitro shoots to rooting media, containing different concentrations of indolebutyric acid (IBA) and indoleacetic acid (IAA), resulted in regeneration of complete plantlets. Alemow and ‘Cleopatra’ mandarin shoots rooted well using these plant growth regulators; however, all combinations of IBA and IAA tested resulted in very low rooting percentages in sour orange. To improve rooting in sour orange and ‘Cleopatra’ mandarin, different combinations of naphthaleneacetic acid (NAA) and IBA were tested. All NAA/IBA combinations produced higher rooting percentages than did the IBA/IAA combinations, and in sour orange nearly 100 % of explants developed roots. An efficient and simple protocol for the micropropagation of three citrus rootstocks, alemow, ‘Cleopatra’ mandarin, and sour orange, by culturing nodes from mature plants, has been established.

Genetic features and surgical managements for von Hippel-Lindau disease type II

Objective To improve the diagnosis and treatment of von H ippel-Lindau (VHL) disease.Methods The clinic data of one kindred with VHL disease type Ⅱ B was reviewed including clinical manifestation,imaging,pathology and therapy.A 40-year-old male complained of the left upper extremity numbness for 6 months,and a variety of visceral tumors were found 3 months ago.Contrast-enhanced MR imaging showed multiple brain tumors.CT showed left kidney tumor,pancreatic tumor,pars-aortic tumor and left adrenal tumor.Fundoscopy showed multiple retinal hemangioblastoma.PET-CT discovered abdominal multiple tumors.And pedigree analysis was to determine the family medical history,and 6 members got genetic screening.Results In the kindred with VHL disease type Ⅱ B,9 members (30％) out of 40members in 4 generations got the disease.Six members got genetic screening,and the result showed 5 (5/6) members had mutation.Three (3/5) members with 3 sites genetic mutation showed clinical manifestation,1 (1/5) members with 3 sites genetic mutation without clinical manifestation found brain tumors by MRI,and 1 ( 1/5 ) member with 1 site genetic mutation did not find disease by comprehensive checkup.The main mutation located at exon 1 in chromosome 3p25 of VHL gene.All mutation was hetcrozygous mutation.The 295,337 and 337 nucleotide thymine of the VHL gene were substituted by cytosine,cytosine and adenine,which made the 98th,112th and 112th tyrosine substituted by histidine and asparagines.One member with 1 site mutation had 98th tyrosine substituted by cytosine.The first operation was to remove brain tumor,and the second operation was to remove adrenal tumor,para-aortic tumor and renal tumor.Pathology of the brain tumors showed hemangioblastoma,and the retroperitoneal tumors were clear cell carcinoma,paraganglioma and pheochromocytoma.Followed up for 6 months after operation,no relapse occurred.Conclusions VHL disease is a relatively rare autosomal dominant disorder.Comprehensive management of patients should also include genetic counseling and screening for other manifestations of the disease process.Genetic testing might be helpful in early detection of asymptomatic VHL patients.Members having gene mutation should be followed up strictly.Surgical management of VHL disease should be decided on the base of comprehensive assess.Multiple lesions could be cut off in one operation.For patients with pheochromocytoma,pheochromocytoma shuld be handled first. Key words: Hippel-Lindau disease; Pedigree; Kindred; Surgical procedures, operative