Objective To observe the changes of intracellular free calcium and the expression of CaM in the hippocampal neurons of posttraumatic stress disorder (PTSD) rats and to further investigate the neurobiological mechanisms. Methods The SPS-method was used to set up the rat PTSD models. A total of sixty male Wistar rats were randomly divided into 12 hours,1 day,4 days,7 days groups of SPS and normal control group. The intracellular free calcium was examined by fluorescence spectrophotometer. The expression of CaM was detected by using immunohistochemistry, Western blotting and RT-PCR. Results The intracellular free calcium level in the hippocampus of experimental rats was markedly increased 12 hours after SPS stimulation,and reached the peak after 1 day, then gradually decreased to normal level after 7 days. The expression of CaM in the hippocampus 1day after SPS was also the highest and then gradually decreased.Conclusion The lasting dysfunction of Ca~(2+)/CaM signaling cascades in hippocampal may play important roles in the pathogenesis of PTSD rats.

By molecular cloning technique, the expressing plasmids pDOR-SV40-Bcl-2 cDNA were successfully constructed. The reconstructed plasmids with lipofectamine were transduced into gastric cancer cell line SGC7901 and then the positive clones which contained the reconstructed plasmids were choosed by G418. Circa 150 positive clones were choosed from 2 x 10_5 gene transduced cells, which suggested that the transducing efficiency was more than 1%o; 2 positive clones were expanded and passage, then 1 drug-resistance cell strain (SGC7901 anBcl-2 cells) was obtained. The bloting results suggested, Bcl-2 cDNA were expressed in both gene transduced and not transduced cell strains, but the expressing level of mRNA and protein in gene transduced cell strain was very low than that in gene not transduced cell strain were positive by the means of Southern blot, Northern blot and Western blot. This results showed that normally in gastric cancer cells the expressing level of Bcl-2 gene was very high, and the cDNA fragment of antisense Bcl-2 were successfully transduced into gastric cancer cells, and in gene transduced cell strains the expressing of Bcl-2 gene was effi-cientlv blocked.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective:To investigate YKL-40-mediated inflammation in human bronchial epithelial cells and analyzed the soluble factors secreted by bronchial epithelial cells exposed to YKL-40 that were responsible for increasing proliferation and migration of primary normal human bronchial smooth muscle cells (BSMCs).Methods:YKL-40-induced inflammation was assayed in two human bronchial epithelial cells (BEAS-2B cell line and primary human bronchial epithelial cells ,namely HBECs).In addition,we treated BEAS-2B cells and HBECs with YKL-40,and added the conditioned culture media ( YKL-40-BEAS-2B-CM) and ( YKL-40-HBECs-CM) to BSMCs.The proliferation and migration of BSMCs were determined by premixed WST-1 cell proliferation reagent and QCM chemotaxis migration assay ,respectively.Results: Bronchial epithelial cells treated with YKL-40 resulted in a significant increase of IL-8 production,but have no effect about RANTES ,Eotaxin and TNF-α.YKL-40-BEAS-2B-CM and YKL-40-HBECs-CM induced IL-8 was found to further stimulate proliferation and migration of BSMCs ,and the effects were inhibited after neutralizing IL-8.Conclusion:Through investigating the interaction of airway epithelium and smooth muscle ,our findings implicate that YKL-40 may be involved in the inflammation of asthma by induction of IL-8 from epithelium,subsequently contributing to BSMCs proliferation and migration.Moreover, inhibition of IL-8 signaling is a potential therapeutic target for YKL-40-induced inflammation and remodeling of asthma.

0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P

Objective To study the relationship between the MDR1 gene expressions and CD56 antigen expression in patients with de novo acute myeloid leukemia(AML) and to explore the role of this two factors in clinical drug resistance and their correlation. Methods A real-time quantitative RT-PCR method was established for detecting MDR1 expression levels and three-color flow cytometry analysis using CD34/ SSC gating was used to examined CD56 antigen expression in 79 de novo AML patients. Results CD56 an-tigen was recorded in 19 out of 79 cases (24.1%) and particularly in those with M5 cytotypes. Moreover, CD56 expression was significantly associated with unfavorable cytogenetic abnormalities (P<0.05), Patients with t(8:21)had a significantly higher incidence (57.1%, 4/7) of CD56 expression than those with favora-ble karyotype(P<0.05). CD56~+ AML patients had a higher incidence of splenohepatomegalia and lactate dehydrogenase level than CD56~- patients(P<0.05). The median expression levels of MDR1 was statistical-ly higher in CD56~+ AML patients than that in CD56 patients(P<0.001). Patients with both high levels of MDR1 and CD56~+ had a significantly lower CR(complete remission) rate than those with both low MDR1 level and CD56 (58.8% vs 89.2%, P<0.01). Conclusion There is a linear correlation between MDR1 gene expression and CD56 expression in AML. Quantification of the MDR1 gene expression together with CD56 antigen expression is more effective to the judgement of prognosis in AML.

AIM: To explore the effect of Snail1 siRNA on high-glucose induced tubular epithelial-to-mesenchymal transition (TEMT). METHODS: Subconfluent renal tubular epithelial cells were incubated in serum-free DMEM for 24 h to arrest and synchronize the cell growth. Then cells were treated with normal glucose (5.5 mmol/L D-glucose) or high glucose (25 mmol/L D-glucose) for 72 h. Meanwhile 19.5 mmol/L D-manntiol was used as high osmotic control. Snail1 siRNA was transfected into tubular epithelial cells. In parallel, cells were transfected with non-specific siRNA which served as the control data sets. Cells were then treated with 25 mmol/L D-glucose for 72 h. RNA and cell lysates were collected to determine the protein and mRNA levels of Snail1, TGF-β_1, α-SMA, vimentin and E-cadherin. RESULTS: Transfection caused the decreases in Snail1 at mRNA and protein levels by 62% and 68% respectively as compared to those in untransfected cells cultured in high glucose medium. Western blotting exhibited that Snail1 siRNA transfection restored E-cadherin protein expression by 61% compared to that in high-glucose-treatment cells, whereas it inhibited high-glucose-induced induction of α-SMA protein by 58%. Similarly, RT-PCR revealed that Snail1 siRNA transfection dramatically suppressed the high-glucose-induced mRNA expressions of α-SMA and vimentin by 72% and 61%, respectively, while E-cadherin mRNA increased by 53%. CONCLUSION: Our study provides direct evidence that Snail1 is able to control TEMT.

Objective To study the protective effect and mechanism of ganciclovir(GCV) on acute cerebral injury of mice caused by herpes simplex virus(HCV). Methods Mice model of acute cerebral injury caused by HCV were established, morphological changes in the brain tissue of mouse treated with GCV were observed under the electronic microscope, and the mortality were compared. The HSV - I DNA copies of brain tissue were detected by fluorescent quantitative polymerase chain reaction. Results In the infected model group, there were obvious swelling, karyopyknosis and destruction of the structure in the brain cells, as well as myelin sheath solution and vacuolar degeneration in the mitochondrion and crest were destroyed. There were the virions in the nucleolus. With the GCV treatment, the symptoms were improved, the mortality much lowered, the yields of HSV - I DNA much lower. Conclusions GCV may restrain replication of HSV-Ⅰ effectively and lower the mortality of mice with acute cerebral injury caused by herpes simplex virus significantly.

A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor (GDNF) on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons (SMNs) were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyltransferase (ChAT) expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.

Objective To investigate the expression and possible mechanism of miR-21 and Ski-related novel protein N( SnoN) in the renal fibrosis diabetic process.Methods The animal model was established by tail-vein injection of Streptozotocin,and the other group were normal control ( NC) group.After 10 weeks, the rats were sacrificed to measure biochemical parameters and renal index , and to observe the changes of pathomorphology by HE staining as well.Meanwhile, immunohistochemistry and Western blot were employed to examine protein ex-pression of E-cadherin,α-smooth muscle actin(α-SMA), fibronectin(FN), collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), transforming growth factor-β1(TGF-β1), Smad3, p-Smad3(Ser423/425) and SnoN in the renal tissue. In addition, the expression of SonN mRNA and miR-21 were detected by qPCR.Results In DM group,the ex-pressions of Col-Ⅰ, Col-Ⅲ and FN in renal interstitium were increased ( P <0.05 ) , TGF-β1 increased (P<0.05),while E-cadherin decreased(P<0.05).Compared with NC group, the expression of α-SMA,p-Smad3 (Ser423/425) protein increased in DM group(P<0.05),while the protein level of SnoN decreased but the level of SnoN mRNA increased ( P <0.05 ) .Moreover, the level of miR-21 markedly increased in DM group ( P <0.05 ) .Conclusions TGF-β1 may up-regulate the expression of miR-21 but restrain the translational expression of SnoN, aggravating fibrosis.