Effects of Huangpu Tongqiao Capsules on EGFR-PLCγ signal pathway of hippocampus in rats with Alzheimer's disease / 中国中药杂志

Huangpu Tongqiao Capsules(HPTQC), with the functions of invigorating Qi and kidney, eliminating phlegm and removing blood stasis, have the effect of treating Alzheimer's disease(AD), but its mechanism needs further exploration. To explore the relationship between the therapeutic mechanism of HPTQC on Alzheimer's disease and EGFR-PLCγ signal pathway, 40 healthy male SD rats were selected and divided into 4 groups randomly: sham operation group(sham), model group(model), HPTQC group(HPTQC), and nimodipine group(NMP). AD rat model was established by intraperitoneal injection of D-galactose combined with an intracerebral injection of amyloid-β peptide(25-35). After 28 days of administration, Morris water maze test and HE staining showed that the learning and memory ability of AD rats were significantly decreased(P<0.01), and hippocampal neurons were obviously da-maged. However, HPTQC could improve the learning and memory ability of AD rats(P<0.05) and reduce the damage of hippocampal neurons. Immunofluorescence test results showed that the expression levels of EGFR and p-Tau in hippocampal CA1 region of AD rats were significantly increased(P<0.01), and HPTQC could reduce the expression of EGFR and p-Tau in hippocampus of AD rats(P<0.01). Western blot results showed that the protein expression levels of EGFR, PLCγ, IP3 R and p-Tau in hippocampus of AD rats were significantly increased(P<0.01), and HPTQC could reduce the protein expression of EGFR, PLCγ, IP3 R and p-Tau in AD rats(P<0.05). RT-PCR results showed that the mRNA levels of EGFR, PLCγ, IP3 R and Tau in hippocampus of AD rats were significantly increased(P<0.01), and HPTQC could reduce the mRNA levels of EGFR, PLCγ, IP3 R and Tau in AD rats(P<0.05). The results indicate that HPTQC can improve the learning and memory ability of AD rats, and its mechanism of action may be related to regulating EGFR-PLCγ signal pathway.

HPPR encodes the hydroxyphenylpyruvate reductase required for the biosynthesis of hydrophilic phenolic acids in Salvia miltiorrhiza / 中国天然药物

Salvia miltiorrhiza is a medicinal plant widely used in the treatment of cardiovascular and cerebrovascular diseases. Hydrophilic phenolic acids, including rosmarinic acid (RA) and lithospermic acid B (LAB), are its primary medicinal ingredients. However, the biosynthetic pathway of RA and LAB in S. miltiorrhiza is still poorly understood. In the present study, we accomplished the isolation and characterization of a novel S. miltiorrhiza Hydroxyphenylpyruvate reductase (HPPR) gene, SmHPPR, which plays an important role in the biosynthesis of RA. SmHPPR contained a putative catalytic domain and a NAD(P)H-binding motif. The recombinant SmHPPR enzyme exhibited high HPPR activity, converting 4-hydroxyphenylpyruvic acid (pHPP) to 4-hydroxyphenyllactic acid (pHPL), and exhibited the highest affinity for substrate 4-hydroxyphenylpyruvate. SmHPPR expression could be induced by various treatments, including SA, GA, MeJA and Ag, and the changes in SmHPPR activity were correlated well with hydrophilic phenolic acid accumulation. SmHPPR was localized in cytoplasm, most likely close to the cytosolic NADPH-dependent hydroxypyruvate reductase active in photorespiration. In addition, the transgenic S. miltiorrhiza hairy roots overexpressing SmHPPR exhibited up to 10-fold increases in the products of hydrophilic phenolic acid pathway. In conclusion, our findings provide a new insight into the synthesis of active pharmaceutical compounds at molecular level.

A case-control study on correlation of pesticide exposure with childhood acute leukemia / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate the correlation of pesticide exposure with childhood acute leukemia.</p><p><b>METHODS</b>An exploratory case-control study was conducted among childhood acute leukemia patients under 15 years old in Shanghai, China. From January 1st, 2006 to December 31st, 2008, a total of 80 newly diagnosed acute leukemia patients were recruited from Shanghai Children's Medical Center for the case group. Another 96 age-matched patients who visited the hospital for health examination, pediatric treatment or osteological therapy excluding hematological system diseases and neoplastic disease, were recruited for the control group. A questionnaire survey was conducted in both groups; and a 30 - 40 ml random urine sample was collected from each participant. Five types of organophosphorus pesticide metabolites was then detected among the samples, using Gas Chromatography with Flame Spectrophotometry.</p><p><b>RESULTS</b>According to result of the questionnaire survey, more participants (55.0% (44/80)) in case group than in the control group (33.3% (32/96)) reported using mosquitocidal, which might increase the risk of childhood acute leukemia (OR = 2.444; 95%CI: 1.326 - 4.506). At the same time, the detection showed that the concentration (median) of organophosphate metabolites diethyl phosphate, dimethyl phosphate, dimethyl thiophosphate, diethyl thiophosphate and diethyl dithiophosphate in case group were 0.0682, 0.0082, 0.0183, 0.0233, 0.4259 µg/g Cr, which were all significantly higher than in control group (0.0865, 0.0025, 0.0112, 0.0123, 0.1207 µg/g Cr) except the concentration of diethyl phosphate (Z = -1.081, P = 0.279). The difference showed statistical significance (Z = -5.752, -2.800, -3.316, -8.120, P < 0.05).</p><p><b>CONCLUSION</b>Pesticide exposure may be one of the risk factors for childhood acute leukemia.</p>

Molecular genetic mechanism of sodium arsenite on growth and development / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of sodium arsenite on gene expression related to growth and development and explored the molecular mechanism of arsenic effects using gene chips.</p><p><b>METHODS</b>Normal human hepatic cells were dripped on chips and then hybrided with the first strand of cDNA from hepatic cell exposed to different concentration of sodium arsenite. Gene sequence of clone differently expressed was determined and then defined which gene it was and finally those genes which associated with growth and development were identified.</p><p><b>RESULTS</b>The p55 gene expression level of two experimental groups was severaly 2.21 and 2.93 times as the control group. The PL gene level of two experimental groups were 0.13 and 0.27 times as the control group, and the HOXA10 gene level was 0.22 and 0.35 times of the control group. These results indicated that sodium arsenite increase p55 gene expression, and inhibited PL and HOXA10 gene expression.</p><p><b>CONCLUSIONS</b>The sodium arsenite could affect the gene expression related to growth and development and it is shown that the molecular genetic mechanism of sodium arsenite is related to growth and development.</p>

Construction of subtracted cDNA library in human Jurkat T cell line induced by arsenic trioxide in vitro / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand the differentially expressed genes in human T lymphocytes induced by arsenic trioxide (As(2)O(3)) and to explore mechanism of its immunotoxicity and immune suppression.</p><p><b>METHODS</b>Human Jurkat T cell line was treated by arsenic trioxide (5 micromol/L, 24 h) in vitro, as a sample model. Then, the differentially expressed genes were cloned and the subtractive cDNA library from Jurkat T cell line was constructed by suppression subtractive hybridization (SSH). Polymerase chain reaction (PCR) and sequencing techniques were applied to identify positive clones.</p><p><b>RESULTS</b>The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was constructed, including 29 different gene fragments and only replicated one in the subtracted cDNA library identified by PCR and sequencing analysis. These gene sequences were 95%-100% analogous to the genes in public database (GenBank/EMBL). The cDNA library contained oxidative metabolic genes in mitochondria (triose phosphate dehydrogenase, NADH4, pyrophosphate synthase, 16S rRNA ribosome, succinate-CoA ligase and ATP synthase 6); transcriptional and translation genes poly (A) binding protein, t-RNA-guanine transglycoslase, ribosomal protein L23, ribosomal protein S15A, eukaryotic translation initiation factor 3, Rab interaction protein 5, splicing factor-arginine serine rich 5, and ADP-ribosylation factor-like 6 interacting protein), oxide stress related genes (ferritin high chain and high-mobility group protein 2); protein activating and signaling pathway related genes (casein kinase, serine kinase 2 and phosphatidylinositol-four-phosphate adaptor protein-1-associated protein); cell differentiation and apoptosis associated genes (NB4 cell apoptosis related protein and myeloid differentiation primary response protein) and five genes with unknown function (KIAA0092, CGI-147protein, GCI-35, nucleolar phosphoprotein Nopp34 and Mus muscular partial mRNA for hypothetical protein), as well as a novel gene unmatched to the sequence in GenBank.</p><p><b>CONCLUSIONS</b>The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was successfully constructed. And, genes not involved in previous research on arsenic were found. Results of analysis for these genetic function suggested that there should be many genes involved in process of T lymphocytes apoptosis or injury induced by arsenic trioxide and that there should still be many genes associated with arsenic that were not reported in the past.</p>

Preparation of albendazole polybutycyanocrylate nanoparticles and study on its pharmaceutical properties and tissue distribution / 药学学报

<p><b>AIM</b>To prepare the target drug delivery systems(TDDS), albendazole polybutycyanocrylate nanoparticles (ABZ-PBCA-NP), its pharmaceutical characters and tissue distributions were simultaneously investigated.</p><p><b>METHODS</b>Albendazole nanoparticles were prepared with the emulsification-polymerization method and the drug-load mechanism of polybutycyanocrylate nanoparticles was studied with the equal-tempaerature adsorption principle. The dialyse dynamic of albendazole from ABZ-PBCA-NP was investigated in four formulations in vitro. The tissue distribution of albendazole in different drug vehicles was studied with isotope labelling experiment.</p><p><b>RESULTS</b>ABZ-PBCA-NP and ABZ-PVP-PBCA-NP fit to the Higuchi and bi-exponent function in vitro respectively. The drug loaded in nanoparticles was abide by the Langmuir adsorption equation. Targeting index of albendazole in liver and spleen in mice are 11.4 and 3.9 after ig 3H-ABZ-PBCA-NP. The bioavailability of albendazole nanoparticle and suspension are 76.0% and 36.9% respectively.</p><p><b>CONCLUSION</b>The absorptive capability of drug was enhance when 4% PVP was added into the nanoparticle, and its release time was lengthen. At the same time, the nanoparticles vehicles increase the albendazole bioavailability.</p>