Objective To observe the effect of iodine deficiency and hypothyroidism on the protein expressions of CREB in the entorhinal cortex of pups.Methods Twenty-eight female Wistar clean rats after pregnancy were randomly divided into control group,hypothyroid 1,2 group and iodine deficient group(7 in each group).From GD6 till PN28,iodine-deficient group was administered with iodine-deficient diet [iodine content:(14.11 ?1.96) ng/g] and tap water;hypothyroid 1 and 2 group were administered with 5 mg/L and 15 mg/L PTU in the drinking water and fed with normal diet [iodine content:(470.50?46.52) ng/g].Control group received tap water and normal diet during the experiment.Five pups from each group were sacrificed and intracardiac perfused at PN7,PN14,PN21,PN28 and PN42.Brains were removed,fixed and sectioned coronally.All sections were observed and were analyzed for the protein expression of CREB by immunohistochemistry in the entorhinal cortex.Results At PN7,there was no significant difference in the expression of Ng in all four groups.At PN14 and PN21,the expression of CREB in the entorhinal cortex in hypothyroid 1,2 group and iodine deficient group were significantly lower than those of controls(P

Objective:To explore the effect of autoimmune regulator to TLRs expressions on peripheral antigen presenting cells(APC).Methods:①pEGFPC3-AIRE plasmid was transfected with liposome.②Confocal microscopy was used to observe the effect of transfection.③RT-PCR assay was used to detect the expressions of AIRE and TLR1-9 in RAW264.7cells at 36,48,72,96 h after transfection.Results:①The plasmid was transfected into RAW264.7 cells successfully,and the efficiency of transfection was 60~70%.②AIRE transfected RAW264.7 cells were achieved,and the best time was 72 h.③At 72 h after transfection,the expressions of TLR1,4,5,9 increased,and TLR3,7,8 reduced.The expression of TLR2,6 increased at 96 h.Conclusion:AIRE may regulate the immune response by control TLR expression in APC.It maintain the effective response to pathogen and tolerance state to self tissues through the effects to different TLRs.

Background and purpose:Drug-resistance is the main obstacle in terms of efficacy of chemotherapy for leukemia, RNA interference(RNAi) strategy possesses the characteristics of specilization, high-efficiency and low-toxicity, and can effectively and specifically inhibit the overexpression of given gene. This study was designed to investigate the effect of small interfering RNA (siRNA) on expression of mdr1 gene and drug-resistance in multidrug-resistant human leukemia K562/ADM cell.Methods:Human multidrug-resistant leukemia cell line K562/ADM over-expressing mdr1 gene was used as the target cells, Two siRNAs (si-mdr1-1 and si-mdr1-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells. Expression of mdr1 mRNA was determined by RT-PCR, P-glycoprotein (P-gp) expression was measured using flow cytometry (FCM), and the sensitivity of K562/ADM cells to adriamycin was assessed with a MTT colorimetric assay.Results:Two siRNAs (si-mdr1-1 and si-mdr1-2) specially designed in this study could markedly down-regulate the expression of mdr1 mRNA and its product P-gp in K562/ADM cells. After cells transfected with si-mdr1-1 or si-mdr1-2 for 24h and 48h, the inhibition of mdr1 mRNA expression in the cells for si-mdr1-1 was 55.5% and 22.5%; and for si-mdr1-2, 16.0% and 57.6%, respectively. Treated with siRNA for 72h, the expression intensity of P-gp in the two transfected cell lines decreased 74% and 85%, respectively. Both si-mdr1-1 and si-mdr1-2 significantly enhanced the sensitivity of K562/ADM cells to adriamycin and reversed their drug-resistance, the reversal efficiency was 2.52-folds and 1.96-folds, respectively.Conclusions:The siRNA could effectively and specifically silence the expression of mdr1 gene and overcome the drug-resistance mediated by P-gp in K562/ADM cells.

Objective To evaluate the activation of premotor area (PMA) in verb generation task, and to discuss the possible function of PMA in language expression. Methods Block-designed fMRI with verb generation task was performed on 23 subjects with GE 1.5T MR Scanner. During the test, the subjects were asked to generate a verb based on a given noun word. The white + appeared on the center of the black screen was used as control. The fMRI data were processed with SPM 2. Group analysis was performed with single sample t-test. Average mapping was obtained and overlapped onto standard MNI template. Activation of the PMA was analyzed. Results The fMRI data of eleven subjects were selected for group analysis after head motion effect was ruled out. Average mapping showed activation in the Broca's area, posterior part of the right inferior frontal gyrus, bilateral PMA and supplemental areas (SMA), left posterior parietal cortex, right thalamus, left basal ganglions, right cerebellum, and posterior part of the right temporal lobe. The area with the greatest activated intensity in the brain was the left PMA. Conclusion PMC is important in verb generation and may be responsible for voice processing, motor imagery, word extracting as well as advanced regulation of information.

Objective To explore the effect of dexmedetomidine on phosphoinositide 3-kinase/protein kinase B (PI3K/Akt ) pathway in hippocampus of propofol anesthetized neonatal rats. Methods Eighty Sprague-Dawley male rats,aged 7 days,weighing 10-1 5 g,were randomly divided into 8 groups (n= 10 each):normal saline group (group N),DMSO group (group D),intralipid group (group I),propofol group (group P),dexmedetomidine 25 μg/kg,50 μg/kg and 75 μg/kg +propofol 100 mg/kg groups (groups PD25 ,PD50 and PD7 5 ),LY294002 25 μg + dexmedetomidine 75μg/kg + propofol 100 mg/kg group (group LYPD).The hippocampus of rats in all groups were taken 2 h after the animals fully awake.The ultrastructure of hippocampal neurons was observed by transmission electron microscope.The pAkt-(ser473 )protein and Akt protein in the hippocampus were evaluated by Western blot analysis.Results There was no significant difference in the expression of Akt protein among the eight groups.Compared with group N,the expression of pAkt (ser473)protein was significantly down-regulated in groups P,PD25 ,PD50 ,PD7 5 and LYPD (P <0.05).Compared with group P,the expression of pAkt (ser473)protein was increased significantly in groups PD7 5 and LYPD (P <0.05).Compared with group PD7 5 ,the expression of pAkt (ser473) protein was significantly down-regulated in group LYPD (P <0.05 ).The structure of hippocampal neurons was normal in groups N,I and D.Nuclear nuclei swelling,chromatin decreasing and mito-chondrion vacuolar degeneration were observed in group P while improved gradually with dexmedeto-midine in a dose-dependent manner in groups PD25 ,PD50 and PD7 5 .Neurons karyopyknosis,partial dissolution of nuclear membrane,chromatin condensation,mitochondria vacuolar degeneration were observed in group LYPD.Conclusion Dexmedetomidine pretreatment provides neuroprotection against propofol-induced hippocampal destruction by preserving PI3K/Akt pathway activity in the de-veloping brains.

AIM:To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms.METHODS:CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay.The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation.The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR,and the protein expressions of c-Myc,Bcl-2,caspase-3 precursor(procaspase-3) and poly ADP-ribose polymerase(PARP) were detected by Western blotting.RESULTS:Baicalin remarkably inhibited the CA46 cell proliferation,with an IC50 value of 10 ?mol/L.Apoptosis was remarkably induced by baicalin in a dose-dependent manner,and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation,respectively.Furthermore,RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner.Western blotting showed that the protein expressions of c-Myc,Bcl-2,procaspase-3 and PARP(116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner,while the expression of PARP(85 kD) was up-regulated.CONCLUSION:Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells,which may be related with the down-regulation of c-Myc and Bcl-2 expressions,as well as the up-regulation of caspase-3 activity.

Objective To investigate whether emodin enhances chemosensitivity of pancreatic cancer cells and the mechanism. Methods Normal human skin fibroblasts and pancreatic BXPC-3 cells were divided into control group, cisplatin (5 mg/L) treatment group, gemcitabine (0.5 μmol/L) treatment group, emodin (50 μmol/L)+ cisplatin (5 mg/L) co-treatment group and emodin (50 μmol/L) + gemcitabine (0.5 μmol/L) co-treatmentgroup. The cell viability was detected by MTT assay, the cell apoptosis rate by flow cytometry, the expression of the multidrug resistance-associated protein 1 ( MRP1 ), MRP2, breast cancer resistance protein (BCRP) and P-glycoprotein (P-gp) mRNA in different cell lines was determined by reverse transcription polymerase chain reaction.All data were analyzed using the t test. Results The cell viability and apoptosis rate of pancreatic BXPC-3 cells after treatment were 62.44% ± 3.42% and 27. 10% ± 4.24% in the cisplatin treatment group, and they were 30.53% ±0.05% and 66.33% ±9.37% in the emodin + cisplatin co-treatment group. There were significant differences in cell viability and apoptosis rate between the two groups (t = 13.20, 5. 35, P < 0.05 ). The cell viability and apoptosis rate of pancreatic BXPC-3 cells after treatment were 79.82% ±2.83% and 13.48% ± 1.65%in the gemcitabine treatment group, and they were 45.65% ± 2.46% and 62.74% ± 10. 18% in the emodin +gemcitabine co-treatment group. There were significant differences in cell viability and apoptosis rate between the two groups (t = 12.89, 8.28, P < 0. 05 ). There was no significant difference in the viability of normal human skin fibroblasts between the cisplatin treatment group and the emodin + cisplatin co-treatment group, and also between the gemcitabine treatment group and the emodin + gemcitabine co-treatment group (t = 2. 08, 0. 64, P >0.05 ). Expression of MRP1 mRNA was detected in pancreatic BXPC-3 cells, whereas the expression of MRP2,BCRP and P-gp mRNA was undetectable. The expression of MRP1 mRNA in pancreatic BXPC-3 cells was significantly down-regulated in the cisplatin treatment group, and cisplatin + emodin co-treatment had an additive effect on down-regulating the expression of MRP1 mRNA. Conclusion Emodin may enhance chemosensitivity of pancreatic cancer cells to cisplatin by down-regulating the expression of MRP1 mRNA.

ObjectiveTo study specific allergic IgG antibody test and its impact factors.MethodsA total of 1770 Qingdao residents were recruited in this cross-sectional study from January to October 2011.ELISA was used to test specific allergic IgG antibodies.Logistic regression was used for multifactor analysis.Results Of 1770 participants,9 individuals had insufficient data,resulting in a response rate of 99.5％.Identification rate of specific allergic IgG antibody was 60.0％ (1056/1761 ),and the most commonly seen allergic foods were crab (49.4％ ),egg (44.80％ ),ling (38.0％ ),shrimp ( 30.4％ ) and soy ( 22.2％ ).Intolerance to crab,egg,shrimp,creamery,com,and tomato showed gender difference ( x2 =18.978,P＜0.05 ).Serum level of allergic IgG antibody was increased with age [20 - 40 years:( 118.61 ±45.67) U/ml; 41-60 years:(166.57 ±55.82) U/ml; ＞60 years:(183.67 ±49.34) U/ml; x2=13.597,P＜0.01].In logistic regression,difference between individuals with normal body weight and obesity population ( OR =1.897,95％ CI 1.215 - 2.578 ) or between those with or without allergic history ( OR =0.780,95％ CI 0.648 - 0.912) was found.Conclusions Impact factor of food intolerance includes gender,age,constitution and body mass index.Six kinds of food intolerance are more common in women.41 -60 year group shows peak of food intolerance.No exposure to allergic food and antianaphylaxis may reduce the risk of food intolerance.

Objective To observe the effect of iodine deficiency and hypothyroidism on the protein expressions of calcineurin in the hip-pocampus of pups. Methods Female Wistar rats (n=28) after pregenancy were randomly divided into control group,hypothyroid group and iodine deficient group. According to the dose of propylthiouracil (PTU) in the fed water, hypothyroid group was divided into 5 ppm group and 15 ppm group (7 rats in each group). Totally 5 pups from each group were sacrificed and perfused intracardially in postnatal day (PN) 7,PN14 and PN21. Brains were removed,fixed and sectioned coronally. All sections were observed and analysed for the protein exression of calcineurin by immunohistochemistry in the hippocampus CA1,CA3 and DG regions. Results In PN14 and PN21,protein levels of cal-cineurin in GA1 and CA3 regions of the hippocampus in iodine-deficient and 15 ppm treatment groups were significantly higher than those of the controls (P< 0.05) and in DG region,the contrary was true. In PN7,the positive products were scarely observated in each region and the protein expression was no significantly different in all four groups. Conclusion Iodine deficiency and hypothyroidism may increase the protein expression of calcineurin.

Objective To explore whether arsenic trioxide(As2O3)-induced apopotosis in drug-resistant leukemia K562/ADM cells may induce in through endoplasmic reticulum stress leukemia cell apopotosis.Methods The apoptosis of K562/ADM cells was identified by double staining of FITC-Annexin V and propidium iodide(PI),the ultrastructure of the cells,endoplasmic reticulum and mitochondria were observed by transmission electron microscopy.Flow cytometry(FCM) was employed to assess mitochondrial inner membrane potential(??m),intracellular calcium concentration,cytochrome c(Cyt c) release and caspase-3 activity.The expression of GRP78 protein was analyzed by Western blot.Results During the apoptotic process of K562/ADM cells induced with 2 ?mol/L and 5 ?mol/L As2O3,the endoplasmic reticulum exhibited obvious expansion and degranulation,and the mitochondria illustrated inner and outer membranes fusion,reduced and confused cristae,swelling and vacuolization.The mitochondrial ??m decreased,the intracellular calcium concentration and releasing of cytochrome c from mitochondria increased,and caspase-3 was activated.Western blot result indicated upregulation of GRP78 protein at endoplas-mic reticulum in apopototic K562/ADM cells.Conclusion As2O3 can initiate the endoplasmic reticulum stress in K562/ADM cells,and induces to apoptosis of the drug-resistant cell via endoplasmic reticulum-mitochondrial pathway.