Objective To explore the developmental toxicity of aluminum sulfate and its mechanism. Methods 8.5-day-old embryos of Kunming mice were explanted and cultured in a whole embryo culture system with Al 3+ concentrations of 0.6, 0.9, 1.2, 3.0, 6.0, 9.0 ?g/ml for 48 h. Each viable embryo was evaluated using Maele-Fabry scoring system, and visceral yolk sac diameter, crown-rump and head length, and embryo dry weight were measured, as well as GSH activity in embryonic tissue by using 5,5-dithion-bis-2-nitrobenzoic acid (DTNB), and membrane lipid fluidity of visceral yolk sac cell by DPH fluorescence polarization technique. Results Al 2(SO 4)3 at Al 3+ concentration of 3 ?g/ml resulted in significant inhibition of development of embryos and differentiation of organs, and increasing prevalance rate of abnormal embryos including open neural tube, small head abnormality and deficit in flexion. At Al 3+ concentration of 6.0 ?g/ml, the activity of GSH and the membrane lipid fluidity of visceral yolk sac decreased significantly. In a certain degree, the dose-effect(response) relationship were observed in the above hazardous effects induced by Al 2(SO 4)3. Conclusion Al 2(SO 4)3 presented potential teratagenicity and embryotoxicity, which might be associated with the decreases of the membrane lipid fluidity of visceral yolk sac and the activity of GSH both induced by Al 2(SO 4)3.

Aim Fuzhisan ( FZS ) , a Chinese herbal complex prescription that has been used for the treat-ment of AD for over 15 years, is known to enhance the cognitive ability in AD patients. In this study, to in-vestigate whether FZS reduces Aβ25-35-induced Tau protein hyperphosphorylation in neonatal rat cortical neurons by suppressing the cyclin-dependent kinase 5 ( CDK5 ) pathway. Methods Neonatal Wistar rats born within 24 h were selected to separate and purify their cortical neurons for culture in vitro. After 7-day culture of cortical neurons in vitro, 20 μmol · L-1 Aβ25-35 was used to act on them for 24 h. Medication groups were pretreated with FZS ( 20 mg · L-1 ) , CDK5 inhibitor roscovitine ( 15 μmol · L-1 ) and cal-pain preparation calpeptin (20 μmol·L-1 ) for 24 h, followed by reaction with 20 μmol·L-1 Aβ25-35 for 24 h. Tau protein phosphorylation levels at Ser396, Ser202 and Thr231 and the protein level of CDK5 acti-vator proteins p25/p35 were assayed by Western blot. Fluorescence intensity was measured with a fluores-cence microplate reader to reflect calpain activity. CDK5 kinase activity was assayed by immunoprecipita-tion. Results After 20 μmol·L-1 Aβ25-35 acting on cortical neurons for 24h, there were increments in the following: Tau protein phosphorylation levels at Ser396, Ser202 and Thr231, CDK5 kinase activity, CDK5 activator protein p25 level, and calpain activity. In the 20 mg·L-1 FZS treatment group, Aβ25-35 was suppressed markedly, resulting in increments in Tau protein phosphorylation levels at Ser396 , Ser202 and Thr231 , suppression of CDK5 kinase activity and p25 protein level, and elevation in calpain activity. Both CDK5 inhibitor roscovitine and calpain preparation cal-peptin, as positive control drugs, also played a role in suppressing Tau protein hyperphosphorylation. Con-clusion FZS can suppress Aβ25-35-induced Tau pro-tein hyperphosphorylation in cortical neurons through the calpain/CDK5 pathway.

Objective　To explore the effects of chemical reaction of aluminum salt in water solution on developmental toxicity.Methods　Inhibiting actions on embryo growth and development,and organ morphogenetic differentiation in rats induced by newly prepared and 11.5-month stored aluminum sulfate solution were observed and compared using whole embryo culture method in rats.Results　Both of the two observed solutions showed a certain embryo toxicity and teratogenicity.The toxicity of stored solution decreased significantly compared with that of newly prepared solution.At the same concentration of aluminum,the total Brown's morphogenetic score reflecting the every index of embryo growth and development and organ morphogenetic differentiation,the incidence rates of terata and the number of dead embryos were significant lower in rats exposed to stored solution compared with those exposed to newly prepared solution,especially the morbidity of embryos.Conclusion　The complexation reaction between aluminum ions (Al3+) and water molecules in stored aluminum sulfate solution resulted in the changes of existing state of Al3+ and its decreasing developmental toxicity to rats.

AIM:Neural stem cells can be induced to differentiate into various types of neural cells such as neurons and neuroglia cells,but the technique of depuration and cultivation does not consummate.This article determines the optimal culture technique of neural stem cells by different culture concentrations and passage methods. METHODS:Experiments were conducted from May to December 2006 at Laboratory of Transplantation Immunity of Sichuan University.①Clean pregnant female rats(embryonic age range from 12-16 days)and the disposition of animal met ethical standard.②The cerebral cortex of rat embryos were collected,and digested with trypsin and ethylenediamine tetraacetic acid mixture to obtain signal cell suspension.They were cultured in serum-free medium (DMEM/F12 medium containing B27,basic fibroblast growth factor and epidermal growth factor).The 3~(rd)passage cells were collected,and incubated at 1?10~7 L~(-1),1?10~8 L~(-1),1?10~9 L~(-1),1?10~(10)L~(-1),respectively.In addition,neural stem cells were collected 7-10 days after primary culture to harvest formative cell masses.Mechanical blow refers to soft blowing with haustorial tube from thick to thin after centrifugation,or sterile syringe with No.5 pinhead blowing cells when the blow was 5 times.Bubble production was avoided during the operation.Trypsin aspiration combined with mechanical blow refers to trypsin was added after centrifugation,at 37℃for 10 minutes,the neural stem cells were lightly blown with haustorial tube polished with flame or blown with sterile syringe with No.5 pinhead,and then fetal bovine serum was added to stop digestion.③The growth characteristic of the 3~(rd)passage cells at different culture concentration was observed and proliferation was measured at days 1,3,5 and 7 by MTT assay.The neural clone spheres of subcultured was counted to determine the optimal passage way.Immunofluorescence was carried out to detect nestin(special marker to neural stem cells),BrdU,neurone specific enolase,glial fibrillary acidic protein. RESULTS:①Growth characteristics and identification of rat embryonic neural stem cells in vitro:The dissociated neural stem cells from the cerebral cortex of rat embryos were continuously harvested and purified by suspension cultures to get the daughter cell clone.Nestin positive cells could be found in the neurospheres and after attachment they could differentiate into neurone specific enolase and glial fibrillary acidic protein positive cells,and immunofluorescence showed a great of BrdU-positive cells.②The effect of different incubated number of neural stem cells on proliferation:When the neural stem cells planted at the concentration of 1?10~9 L~(-1),the growth rate of the cells was the highest of all concentrations.The number of clone spheres exceeded others at concentration 1?10~7 L~(-1),1?10~8 L~(-1)and 1?10~10 L~(-1)(P

Objective To explore the effects of chemical reaction of aluminum salt in water solution on developmental toxicity Methods Inhibiting actions on embryo growth and development,and organ morphogenetic differentiation in rats induced by newly prepared and 11 5 month stored aluminum sulfate solution were observed and compared using whole embryo culture method in rats Results Both of the two observed solutions showed a certain embryo toxicity and teratogenicity The toxicity of stored solution decreased significantly compared with that of newly prepared solution At the same concentration of aluminum,the total Brown's morphogenetic score reflecting the every index of embryo growth and development and organ morphogenetic differentiation,the incidence rates of terata and the number of dead embryos were significant lower in rats exposed to stored solution compared with those exposed to newly prepared solution,especially the morbidity of embryos Conclusion The complexation reaction between aluminum ions (Al 3+ ) and water molecules in stored aluminum sulfate solution resulted in the changes of existing state of Al 3+ and its decreasing developmental toxicity to rats

Objective To study the toxic effect of flutamide (Flu) on testis with DNA chip. Methods Flu was injected subcutaneously at doses of 6.25 mg?kg-1 ?d-1 to 25 F0 rats from gestation day (GD) 12 to 17, and the 25 rats of control group received same doses of normal saline. All 50 pregnant SD rats were sacrificed on GD 20.The cDNA were extracted respectively from fetal testis, then reversely transcribed to cDNA and labeled with cy5 and cy3 fluorescence. Subsequently, cDNA probes were hybridized to the Mouse40S cDNA microarray and fluorescence signals were analyzed. Testosterone level was determined by enzyme-linked immunosorbent assay (ELISA). Results Compared with control,a total of 31 identified gene was found to be expressed differently, including 14 down-regulated and 17 up-regulated. Among them, 5 down-regulated and 10 up-regulated genes were unknown in Genbank. ELISA indicated that testosterone levels were decreased in Flu toxic group. Conclusion Flu treatment to F0 pregnant rats can damage apparently to the reproductive development of embryonic period male SD rats. The obtained differential genes may provide reference for the molecular mechanism of reproductive development.

AIM: To study extraction process with the aid of cyclodextrin (EPAC) of Radix Salvias Miltiorrhizae. METHODS: Taking tanshinone Ⅱ_A, Danshensu, dry extract as the indexes, EPAC and water extraction process(WEP) were compared. RESULTS:The index of tanshinone Ⅱ_A of EPAC was better than WEP in extract, but there was not obvious difference in the indexes of Danshensu and dry extract. CONCLUSION:EPAC is worth researching further.

Objective To research the absorption mechanism of paeoniflorin across Caco-2 monolayer model. Methods Depending on the Caco-2 cell monolayers drug transport model to study the double transport mechanism of paeoniflorin. To explore the effect of paeoniflorin absorption by time and drug concentration, the drug concentration was determined through the use of HPLC instrument and the Papp was calcalatad. Results In the Caco-2 monolayer model, the transport of paeoniflorin form Apical (AP) to Basolateral (BL) is similar to the transport form BL to AP. Conclusion The main mechanism of the paeoniflorin intestinal absorption in the Caco-2 monolayer model is passive transference.

8 ?mol/L)was markedly restrained. Zearalenone, like estradiol, could markedly increase the proliferation of MCF-7 cells. The effects were time-dependent and dose-dependent. Conclusion: The tested phytoestrogens are biologically active, and they can differently affect the proliferation of human breast cancer cells in vitro. These data suggest that genistein may have preventive and therapeutic applications against breast tumors.

Objective: This study was designed to investigate the molecular mechanisms of proliferation and apoptosis by genistein and zearalenone through regulation of mRNA and protein expression of PCNA, bcl-2 and bax in breast cancer MCF-7 cells. Methods: The cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments, the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran–treated FBS. The cells were harvested and seeded in 6-well culture plates or in 75 ml flacks. After various concentrations of genistein and zearalenone treatments for 72 h, the cells were harvested and mRNA and protein expression of proliferation cell nuclear antigen(PCNA), bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. Results: At the concentration of 75 ?mol/L, GS could significantly down-regulate bcl-2 and PCNA mRNA expression and up-regulate bax mRNA expression, and zearalenone indicated an opposite result. These results were further confirmed by following immunohistochemistry. Conclusion:PCNA, bcl-2 and bax pathway might be involved in cell proliferation and apoptosis events regulated by dietary estrogens genistein and zearalenone in breast cancer MCF-7 cells.