Study on Insecticidal Activities and Effect on Three Kinds of Enzymes by 5-Aminolevulinic Acid on Oxya chinensis

Insecticidal activities and effects on three enzymic activities caused by 5-aminolevulinic acid （ALA） on Oxya chinensis were studied. Fourth-instar nymphs of O. chinensis were treated with different doses ofALA （A1,250 mM; A2, 450 mM; A3,750 mM; A4, 1 000 mM）. Mortality and the activities of acetylcholinesterase （ACHE）, glutathione S-transferase （GSTs）, and glutathione peroxidase （GPx） were determinated. The mortality of O. chinensis rose with an increasing dose of ALA. The mortality of high-dose treatments A3 and A4 reached 66.19 and 80.21%, respectively. The value of LD50 was 3.61 （3.29-3.93） mg·g^-1 body weight （95% confidence interval）. Biochemical studies showed that the activities of AChE and GPx in the A4 treatment declined by 51.53 and 42.82% in the female, and 42.65 and 43.85% in the male compared to the control, respectively, and the degree of decline reached a significant level at P 〈 0.05. Meanwhile, the GSTs activities of O. chinensis enhanced with increasing dose of ALA. The GSTs activities of female and male O. chinensis in the A4 treatment remarkably increased by 171.05 and 97.42% compared to the control （P〈 0.05）. ALA had an obviously toxic effect on O. chinensis. Moreover, ALA caused the photoinactivation of AChE and GPx, which induced nerve transmission blocking and the capability to defend oxidation damage declining. Meanwhile, a high dose of ALA could activate GSTs, which caused a feedback inhibition of the insect to the phototoxic substance.

OPTIMIZATIONS FOR 5-AMINOLEVULINIC ACID BASED PHOTODYNAMIC THERAPY IN PURGING LEUKEMIA CELL HL60

Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.

套袋和外源5-氨基乙酰丙酸处理对‘云红梨2号’果皮着色的影响

以‘云红梨2号’为试材,研究套袋、外源5-氨基乙酰丙酸(ALA)处理对果皮花色素苷含量、花色苷合成关键酶活性、可溶性总糖含量以及内源ALA含量变化的影响,并探讨果皮花色素苷含量与酶活性及可溶性总糖含量的关系。结果表明:在果实着色初期,套袋和外源ALA处理均有明显的促进果皮着色的效果,且两者共同处理促进作用更为显著;随着果实接近成熟,花色素苷含量有所下降。在正常光照情况下,果实摘袋后5 d左右对促进着色效果较好。相关性分析表明,套袋处理的果皮花色素苷含量与苯丙氨酸解氨酶(PAL)活性显著相关;不同处理及对照果皮花色素苷含量均与类黄酮3-O-葡萄糖基转移酶(UFGT)活性显著相关,但与果实可溶性总糖含量无显著相关性。

Promotion by 5-Aminolevulinic Acid of Germination of Pakchoi （Brassica campestris ssp. chinensis var. communis Tsen et Lee） Seeds Under Salt Stress

The seed germination and seedling growth of pakchoi （Brassica campestris ssp. chinensis var. communis Tsen et Lee cv. Hanxiao） were not significantly inhibited until the concentration of NaCl was increased to 150 mmol/L. Treatment of pakchoi seeds with exogenous 5-aminolevulinic acid （ALA）, at concentrations ranging from 0.01 to 10.00 mg/L, promoted seed germination when seeds were stressed by salinity, whereas levulinic acid （LA）, an inhibitor of ALA dehydrase, significantly inhibited seed germination and seedling growth, suggesting that metabolism of ALA into porphyrin compounds was necessary for seed germination and seedling growth. Determination of respiratory rate during seed germination showed that ALA increased seed respiration under both normal conditions and salt stress. Furthermore, salt stress decreased levels of endogenous ALA, as well as heme, in etiolated seedlings. More salt-tolerant cultivars of pakchoi contained higher relative levels of endogenous ALA and heme under conditions of salt stress. These results indicate that salt stress may inhibit the biosynthesis of endogenous ALA and then heme, which is necessary for seed germination, and treatment of seeds with exogenous ALA prior to germination may be associated with the biosynthesis of heme.

Study of possibility of cell recognition in brain tumors

The brain has an exceptionally high requirement for energy metabolism,with glucose serving as the exclusive energy source.Cancers,including glioblastoma,have a high glucose uptake and rely on aerobic glycolysis for energy metabolism.The alternation of high-efficiency oxidative phosphorylation to a low-efficiency aerobic glycolysis pathway(Warburg effect)provides macromolecules for biosynthesis and proliferation.Current research indicates that the specific metabolism in the tumor tissue and nonnal brain tissue in the glioma allows the use of 5-aminolevulinic acid(5 ALA)-induced protoporphyrin IX(PpIX)and methylene blue(MB)to monitor and correct the development of the tumor.The focus is on the detection of the differences between tumor cells and tumorassociated macrophages/microglia using spectroscopic and microscopic methods,based on the fluorescent signals and the difference in the drug accumulation of photosensitizers(PSs).Since 5 ALA has long been used effectively in the clinic for fluorescent surgical navigation,it was employed as an agent to identify the localization of tumor tissue and study its composition,particularly tumor and immune cells(macrophages),which have also been shown to actively accumulate PpIX.However,since PpIX is photodynamically active,it can be considered effective as the main target of tumor tissue for further successful photodynamic therapy.MB was employed to visualize resident microglia,which is important for their activation/deactivation to prevent the reprogramming of the immune cells by the tumor.Thus,using two drugs,it is possible to prevent crosstalk between tumor cells and the immune cells of different geneses.