Drosophila models for studying iron-related neurodegenerative diseases / 生理学报

In recent years, iron has been regarded as a common pathological feature of many neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and Friedreich's ataxia (FRDA). A number of genes involved in iron transport, storage and regulation have been found associated with initiation and progression of neurodegeneration. However, whether iron abnormalities represent a primary or secondary event still remains unknown. Due to the limitation in transgenic rodent model construction and transfection systems, the progress in unraveling the pathogenic role of different iron-related proteins in neurodegenerative diseases has been slow. Drosophila melanogaster, a simple organism which has a shorter lifespan and smaller genome with many conserved genes, and captures many features of human nervous system and neurodegeneration, may help speed up the progress. The characteristics that spatial- and temporal-specific transgenic Drosophila can be easily constructed and raised in large quantity with phenotype easily determined turn Drosophila into an excellent in vivo genetic system for screening iron-related modifiers in different neurodegenerative conditions and hence provide a better picture about the pathogenic contribution of different iron-related protein abnormalities. It is believed that identification of important iron-related genes that can largely stop or even reverse degenerative process in Drosophila models may lead to development of novel therapeutic strategies against neurodegenerative diseases.

Increased intracellular iron and mineralization of cultured hFOB 1.19 cells following hepcidin activation through ferroportin-1

To address whether hepcidin functions in bone metabolism. This study was carried out in the Laboratory of Radiation Medicine and Public Health of Soochow University, and the Laboratory of the Second Affiliated Hospital of Soochow University, Suzhou, China, from September 2009 to July 2010. The positive expression of ferroportin-1 [Fpn-1] was detected by reverse transcrptase-polymerase chain reaction. After the treatment with distilled water [control group] and hepcidin [25noml/L, 50noml/L, 100noml/L], the fluorescence intensity related to intracellular iron concentration of a human fetal osteoblast cell line [hFOB 1.19] was measured by a confocal laser scanning microscope. A 3-[4,5- dimethylthiazol-2-yl] - 2-5-diphenyltetrazolium bromide assay, and Von Kossa staining was performed to evaluate cell proliferation and mineralization in cultured hFOB 1.19 cells. This study revealed a high level expression of Fpn-1 in hFOB 1.19. On the basis of which, it was found that 25noml/L, 50noml/L, 100noml/L hepcidin could promote the fluorescence intensity related to intracellular iron concentration and mineralization in hFOB 1.19 in a dose-dependent manner [p<0.05], but hepcidin had no effect on FOB 1.19 proliferation [p>0.05]. The hepcidin-ferroportin signal pathway may function in the osteoblast cell line of hFOB 1.19 cells. It is also suggested that cross-talk between iron and calcium homeostasis may play a role in bone metabolism in responding to hepcidin activation

Determination of butylidenephthalide in Ligusticum chuanxiong by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To determine butylidenephthalide in Ligusticum Chuanxiong with RP-HPLC.</p><p><b>METHOD</b>The sample was extracted with methanol using sonication. The ESTD was used to quantify butylidenephthalide. HPLC separation was carried out in a Hypersil ODS columm (4.6 mm x 150 mm, 5 microm) , eluted at 1 mL x min(-1) with methanol-5% isopropyl alcohol (60: 40) at 25 degrees C. The detection wavelength was 230 nm.</p><p><b>RESULT</b>The linear range was 0.07-0.7 microg for butylidenephthalide. The average recovery was 95.3%, and RSD was 2.3% (n =6).</p><p><b>CONCLUSION</b>This method was simple and could be used to determine butylidenephthalide with satisfactory accuracy and reproducibility.</p>

Experimental study of effect of tanshinone on artery restenosis in rat carotid injury model / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the preventive and therapeutic effect of tanshinone (TA) on artery restenosis in the rat carotid injury model and explor the mechanism.</p><p><b>METHOD</b>Male SD rats were randomly divided into model control group, and low dose, moderate dose and high dose TA groups. Each group had 10 rats. The rats in the high, moderate and low dose groups were respectively fed with TA 120, 40,13.3 mg x kg(-1) x d(-1) by gast rogavage; the rats in the model control group were fed with the same volume solvent. Two days later, the rat's right carotid artery was injuried by balloon dilatation to induce intimal thickening for establishing the restenosis model. After 2 weeks of treatment, the artery was harvested and stained by hematoxylin-elsin (HE) and immunohistochemistry of PCNA, NF-kappaB and iNOS. The morphological changes were checked under microscope. The area of the intimal and medial layer of the vessels, and their ratios were analyzed with image analysis software. The expression level of PCNA, NF-kappaB and iNOS were used as the positive index.</p><p><b>RESULT</b>The intimal area and intima-to-media ratio of the injuried artery increased obviously, suggesting the model was successful. Compared with the model group, TA significantly decreased the intimal area and intima-to-media ratio (P < 0.05), and also decreased the positive index of PCNA and the positive ratio of NF-kappaB and iNOS (P < 0.05).</p><p><b>CONCLUSION</b>TA can effectively inhibit intimal thickening and inflammation. This result suggestes that TA may play a positive role in the prevention of restenosis after PTCA.</p>

The new progress of the study about volatile oil of the angelica / 中国中药杂志

To summarize the new progress of the study about volatile oil of the angelica, including the distillable methods, the analysis of the chemical components, the pharmacological effects and the clinical applications. We tracked and searched the correlative references and study reports about volatile oil of the angelica in CNKI data base(1994-2004) and Medline data base (1997-2004). We summarized and compared the different distillable methods of volatile oil of the angelica, meanwhile we summarized many study reports about the analysis of the chemical components of volatile oil of the angelica and it's pharmacological effects, including the toxicity of the volatile oil and it's effects on the uterus smooth muscle, cardiovascular system, respiratory system, central nerve system and immune system. Finally we summarized the clinical application of the volatile oil of the angelica. There are three distillable methods of volatile oil of the angelica . The harvest efficiency of volatile oil is different with different distillable methods. The chemical components are very complicated and the new chemical components are separated and identified. The volatile oil has bidirectional effects on the uterus smooth muscle. It can inhibit the contraction of the uterus smooth muscle induced by different mechanisms. Meanwhile it can depress the blood pressure and ameliorate the cardiac ischemia. The volatile oil can resist the arrhythmia and asthma, restrain the central system, improve the immune function. Nowadays the volatile oil of the angelica is applied to therapy the dysmenorrhea and disorder of the catamenia. The chemical components of the volatile oil of the angelica are very complicated, moreover the pharmacological effects of the volatile oil are comprehensive. People make the new progress of the study about volatile oil of the angelica.

Baroreceptor reflex in heart failure / 生理学报

Congestive heart failure is a syndrome that is usually initiated by a reduction in pump function of the heart, i.e. a decrease in cardiac output. Initially, a reduction in cardiac output leads to unloading of baroreceptor reflex that, in turn, increases heart rate through vago-sympathetic mechanisms and total peripheral resistance via an increase in sympathetic outflow to vascular beds. In this review we are thinking on how baroreceptor reflex plays a role in the abnormal control of the circulation in heart failure. This review and our recent studies suggest that: (1) baroreceptor reflex is blunted in heart failure; (2) central angiotensin II and reactive oxygen species play an important role in blunted baroreceptor reflex; (3) cardiac sympathetic afferent stimulation and chemoreceptor reflex inhibit baroreceptor reflex; and (4) exercise training normalizes abnormal reflexes in the heart failure state.

The experimental study of the effect of tanshinone on artery restenosis in mouse / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the preventive and therapeutic effect of tanshinone (TA) on artery restenosis in mice and primarily explore the mechanism.</p><p><b>METHOD</b>Female KM mice were randomly divided into model control, low dose and high dose TA groups. Each group had 12 mice. The low and high dose drug groups were respectively given TA 3 and 6 g x kg(-1) x d(-1) by ig; the model control group was given the same volume solvent. The controlateral carotid of ligated artery of model control group was regarded as normal control. 2 days later, the mice' s left common carotid artery was dissected and ligated near the carotid bifurcation, leading to intima hyperplasia and then establishing restenosis model. 4 weeks later, the artery was harvested and stained by hematoxylin-elsin (HE) and immunohistochemistry of PCNA. The morphological changes were checked under microscope; the area of the intimal and medial layer of the vessels, and their ratios were analyzed with image analysis software. The expression level of PCNA was expressed as the positive index.</p><p><b>RESULT</b>Compared with those of normal artery, the intimal area, media area and intima-to-media ratio of ligated artery increased obviously (P < 0.01). But TA could significantly decrease all of these parameters (P < 0.01), and also decrease the positive index of PCNA (P < 0.01).</p><p><b>CONCLUSION</b>TA effectively inhibits intima hyperplasia, which is mainly characterized with the proliferation and migration of smooth muscle cell induced by abnormal hemodynamic changes. This result suggestes that TA may play a positive role in the prevention of restenosis after PTCA.</p>

Increased DMT1 expression and iron content in MPTP-treated C57BL/6 mice / 生理学报

Iron plays a key role in Parkinson s disease (PD). To illustrate the mechanism underlying the increase of iron in substantia nigra (SN) in PD, changes of the expression of divalent metal transporter 1 (DMT1) and iron content were examined in SN in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated mice using immunohistochemistry and histochemistry respectively. Following MPTP treatment for 3 d, elevated iron staining was found in SN. A further increase in iron content was observed after 7 d. In these lesioned animals, tyrosine hydroxylase-immunoreactive DA neurons exhibited a decrease in number and morphological changes as well. There were two isoforms of DMT1 expressed in SN of mice. After MPTP treatment, the expression of DMT1 without IRE form increased in either group, whereas DMT1 with IRE form increased only after 7 d of MPTP treatment. These observations suggest that DMT1 is possibly involved in the process of iron accumulation in SN of MPTP-treated mice, which might be responsible for the subsequent death of DA neurons.

Developmental expression and regulation of divalent metal transporter 1 in rat heart / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of divalent metal transporter 1 (DMT1) mRNA in male Sprague-Dawley rat heart of different ages and the expression of DMT1 regulated by dietary iron.</p><p><b>METHODS</b>Reverse transcriptase (RT)-PCR and Western blot were used in this study.</p><p><b>RESULTS</b>(1)Two isoforms of DMT1 mRNA [with and without iron-responsive element (IRE)] were both detected in rat heart, which were correlated with heart iron content. During development, both of two isoforms of DMT1 mRNA expression were the lowest at the age of PND 7, and increased at PND 21, 63 to 196. (2) After fed with a high iron diet or low iron diet for 6 weeks, the rats developed iron overload or iron deficiency respectively. No significant differences in DMT1 mRNA expression were detected among iron overload, iron deficiency and control rats. By using Western blot analysis, a 21% and 40% reduction in DMT1 protein non IRE form and IRE form respectively were found in iron overload rat (P<0.01, compared with control). Increases (26% approximate, equals 28%) in the levels of two isoforms of DMT1 protein were also observed in iron deficient rat (P<0.01, compared with control).</p><p><b>CONCLUSION</b>The level of DMT1 mRNA expression in heart is age dependent;the two isoforms of DMT1 protein may be both regulated by iron on the posttranscriptional mechanism.</p>

Trehalose Production by Permeabilized Cells / 微生物学通报

A permeabilization method of Micrococcus cells has been established. The obtained penneabilized cells could be used for several batches and in the mean time the intracellular enzymes still keep high activity. This method will undoubtedly increase the productivity of the cells and cut down the cost; therefore, it will be a promising way in the treha-lose industrialization. The experiment shows: after being treated with 5% (v/v) methylbenzene for 40min, the cells in suspension were converted into permeabilized cells, then the latter acted on 10% liquefied starch to produce trehaloae, the conversion ratio could reach 70%. The penneabilized cells could be used at least for 6 batchs (12h per batch) and the intracellular enzyme activity kept steady.

HE EFFECT OF MYCELIAL MORPHOLOGY ON LYCOPENE FERMENTATON / 微生物学通报

Mycelial morphology are exploited a great influnence to the metabolites of fungi. The effect of the mycelial morphology on producing lycopene by Blakeslea tripora was investigated. The results indicated that pellets has little role on fermentation when adding kerosene and triton-x100, However, The content of lycopene in the medium reached to 98.6mg/L by adding Ig/L span-20, forming dispersed mycelia, which is 3 times of the control experiments.

THE EFFECT OF IMMOBILIZATION PROCESS TO THE ACTIVITY OF MTHASE AND MTSASE / 微生物学通报

Amylum can be transformed into trehalose on the action of the MTSase and MTHase, however, the percentage of transformation is low. The research on the enzyme immobilization through four kinds of immobilization carrier is described in this paper. The result shows that the carriers, which are put into with enzyme liquid and crossbonded with chi-tosan and glutaraldehyde at first, can adsorb many enzymes independent from this process of trehalose synthesizing, consequently, the enzyme activity is increased sharply. By comparing with the influence of the immobilizing process and some factors in the reaction conditions, such optimize condition can be reached: the time for carrier crossbonding with glutaraldehyde is 18hours , the time with enzyme is another 18hours , and the time of reaction with amylum (10%) is 9hours.The result of the content of trehalose can reach 27.22g/L and the percentage of transformation can reach 54.43% , which previously is 2.67g/L and 5.33% respectively before dealt with by carriers.