Table of Content

05 June 2011, Volume 31 Issue 6

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Influence of P.yoelii MIF on mouse spleen CD8+DC secreting cytokines

En-peng LIU Ming-yue LUO Ding-ding SHAO Heng WANG

2011, 31(6):  597-601. 

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Objective To investigate the effects of MmMIF and PyMIF in vitro on the TLR4 expression, the Surface Molecules Expression Level and IL-12, TGF-β1 production of Spleen DC. Methods Recombinant plasmids were transformed into Escherichia coli BL21 (DE3), and expression of the recombinant PyMIF-His and MuMIF-His was induced by the addition of 0.5 mM IPTG to the bacterial culture and incubated for 4 h at 37°C. The fusion protein was purified using Nickel-affinity chromatography column and endotoxin was removed from the recombinant protein by Sep-Pak C8 column. CD4+DC /CD8+DC were collected from Spleen by CD4+DC /CD8+DC Isolation Kit. After 24 h of treatment with LPS, MmMIF or PyMIF, the TLR4 expression and the surface molecules expression level of CD4+DC and CD8+DC were determined by flow cytometry. IL-12, TGF-β1 production were detected by ELISA. Results The surface molecules expression level of DC didn't change. PyMIF treatment of CD8+DC down-regulated surface expression of TLR4 and IL-12 (from 433±48 pg/mL to 373±30 pg/mL, P<0.05) secretion decreased significantly after LPS stimulation. PyMIF treatment of CD8+DC increased TGF-β1 (from 136±4 pg/mL to 182±7 pg/mL, P<0.05) significantly. Conclusion PyMIF treatment exerts no effects on maturation of Spleen DC. CD8+DC modified by PyMIF down-regulated IL-12 significantly after LPS stimulation and immature CD8+DC modified by PyMIF up-regulated TGF-β1 secretion, thereby regulating the host immune response and conducive to P. yoelii survival.

The effect of Hax1 gene and its specific small interference RNAs on the apoptosis of 293 cells

Lin LONG Li LIU

2011, 31(6):  602-608. 

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Objective To construct the Hax1 gene specific siRNA (sihax), to detect sihax mediated Hax1 gene repression, and to study the influence of Hax1 and Hax1 siRNAs on cell apoptosis. Method The oligo sequences of Hax1 siRNAs are selected, and the constructed eukaryotic expression vectors are transfected into 293 cells. The influence of Hax1 and its siRNAs on the cell apoptosis is evaluated by a number of approaches such as inverted fluorescence microscope, flow cytometric analysis, annexinV-EGFP/PI double staining, and standard RT-PCR. Results Hax1 specific siRNAs, pBS/U6-sihaxA and pBS/U6-sihaxB, have been successfully constructed. Analysis by RT-PCR and inverted fluorescence microscope demonstrate that the inhibitory effect induced by sihaxB is stronger than that of sihaxA. Over-expression of Hax1 significantly inhibits the early apoptosis of 293 cells. Inhibiting the endogenous Hax1 gene expression by by sihaxA and sihaxB markedly increases late apoptotic cells from 9.03?0.473% to10.8?0.513% (p<0.05) and 16.6?0.858% (p<0.01), respectively. In the same time, sihaxB also can reduce the early apoptotic cells from 37.3?0.5% to 32?1.77% (p<0.01). Conclusion The sequence conservation of the targeting site might influence the inhibitory effect induced by Hax1 siRNA. Hax1 gene might actively involve in the multiple regulatory processes related to cell apoptosis. This study provides some valuable information for the functional study on Hax1 gene in the future.

Construction of recombinant lentiviral vector of MicroRNA-29b and its inhibition on the migration and proliferation of gastric cancer cells

Chong-liang JIANG Xiao-yan WANG Jia-nan GONG Fang WANG Jia YU Tao FENG

2011, 31(6):  609-614. 

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Objective: To construct the recombinant lentivirus vector expressing hsa-miR-29b and observe the effect of over-expression of miR-29b on gastric cancer cell migration and proliferation. Methods The pMIR-miR-29b lentivirus vector was generated by PCR amplification of miR-29b precursor sequence. 293TN cells were transfected with pMIR-miR-29b、Packaging Plasmid(s) and pVSV-G plasmids to package the recombinant lentivirus. HGC-27 cells were infected with recombinant lentivirus and the expression level of miR-29b was measured by real-time PCR.The expression of target gene MCL-1 was tested by western blot .Scratch test and CCK8 were used to observe the change in cell migration and proliferation after miR-29b was over-expressed in gastric cancer cells. Results The expression vector of 29b restructuring slow virus have been successfully achieved. After infected stomach cancer cell HGC-27, successfully achieved the overexpression of miR-27. The miR-29b that is overexpressed can inhibit the expression of target gene MCL-1 in HGC-27, and inhibit the ability of migration and proliferation of HGC-27. comprised with untreated group and Lenti-vector group, the migration ability of Lenti-29b group is reduced significantly (P<0.01); comprised with Lenti-vector group, the Proliferative capacity of lenti-29b group is reduced significantly at 48h,72h and 96h(P<0.01). Conclusion Restructuring lentivirus particles that has over expression of miR-29b have been successful achieved. MiR-29b can inhibit the expression of target gene MCL-1 and the migration and proliferation of HGC-27.

Is PKM2 a ligand of TCRγδ?

Jing-fei SHI Lian-xian CUI Wei HE

2011, 31(6):  615-620. 

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Objective To check whether pyruvate kinase isoenzyme type M2 (PKM2) is the TCRγδ’s ligand. Methods By Western blot, the binding of OT3 peptide (the CDR3 region of the δ chain in TCRγδ) with PKM2 was analyzed; with Western blot and immunohistochemistry, the expression of PKM2 in tumor cells and tissues was observed; by flow cytometry and confocal method, PKM2’s location in the tumor cell was tested. The test of immobilized PKM2 expanding γδT cells in vitro and the detection of IFN-γ secreted by PKM2-stimulated γδT cells were used to observe the function of PKM2. Results 1. The binding of PKM2 with OT3 peptide and containing a large number of PKM2 in tumor cells hints that the catching of PKM2 by OT3 peptide from total protein of tumor cells extracts is reasonable and that finding OT3 binding proteins by this method is feasible; 2. PKM2 is widely expressed in tumor cells and tissues, but not expressed on the tumor cell membrane; 3. PKM2 neither expands γδT cells nor stimulates the secretion of IFN-γ from γδT cells in vitro. Conclusion PKM2 is not expressed on the surface of tumor cells, and can’t stimulate activation of γδT cells. PKM2 does not have properties of TCRγδ ligand.

The regulation of carboxyamidotriazole (CAI) on inflammatory factors of BV2 cell model

2011, 31(6):  621-624. 

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Objective This study is designed to explore the regulation of carboxyamidotriazole (CAI) on inflammatory factors of BV2 cell model Methods BV2 cells were incubated with different concentrations of CAI(5~40μmol/L), LPS was then added and cells were incubated for 18 h, TNF-α, IL-1β, IL-6 levels were determined with ELISA kits and NO levels were determined with Griess reagent, respectively the mRNA levels of TNF-α, IL-1β, COX-2 and iNOS were detected by Real-time Quantitative PCR Results The viability of BV2 cells was not affected at the concentrations of CAI used (5~40μmol/L) , CAI was found to reduce the levels of TNF-α products (359.4 ±12.9 vs241.6 ±16.1*,135.7 ±6.4**,5.3 ±1.9**pg/ml, p<0.01) and IL-1β, NO, IL-6 products (p<0.01) in a dose-depended manner at the concentrations 20,30 and 40μmol/L; CAI was also found to inhibit the mRNA expression of TNF-α，IL-1β，COX-2 and iNOS Conclusion The present study suggests that CAI has obvious effect on TNF-α，IL-1β，NO，IL-6 secretions of BV2 cells model, and CAI was found to has a remarkable inhibitory effect on the activation and inflammatory response of the LPS-induced BV2 cells

PARG silencing inhibits lymphangiogenesis in mouse colonal carcinoma in vitro

Wei-qiang WANG Ya-lan WANG Juan PAN Jia-xin YAN

2011, 31(6):  625-629. 

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Objective To investigate the influence of PARG silencing on the tumor lymphangiogenesis. Methods Lentiviral-PARG-shRNA was transfected into mouse colon carcinoma CT26 cell line. The CT26 cells stably expressing PARG gene were established by puromycin selection. The expressions of PARG, PARP-1, NF-κB and VEGF-C protein were detected by Western blot analysis. BALB/c mice benign lymphangiomas in abdominal cavity were induced by injection of 0.2ml incomplete Freund’s adjuvant (IFA, 1:1 with PBS) and mechanically disrupted to obtain LECs. Expressions of VEGFR-3 and Podoplanin were analyzed by immunofluorescence cytochemistry. The capability of LECs to form lymphatic vessel-like structures was detected by a LEC-CT26 cells co-culture method. Results The expression levels of PARP-1, NF-κB, and VEGF-C were reduced in PARG-silencing CT26 cells (P<0.05). Benign lyphangiomas were successfully induced by emulsified IFA. The expressions of VEGFR-3 and Podoplanin were positive in isolated LECs. The lymphatic vessel-like structures in PARG-silencing CT26 cells were significantly less than that in the control group (P < 0.05).Conclusion These studies demonstrate that PARG silencing significantly reduces VEGF-C expression and suppresses the formation of lymphatic vessel-like structures. We therefore propose that PARG silencing has an antilymphangiogentic effect and probably prevent the metastatic dissemination by lymph system.

Culture of mouse cortex neural stem cell and miRNAs’ influence on NSCs’ differentiation ability

Xiang-yu ZHAO Peng-cheng SHU Jing ZHANG Bin YIN Xiao-zhong PENG

2011, 31(6):  630-635. 

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Objective To establish the mouse cortex neural stem cells (NSCs) culture technology platform in vitro and to explore some miRNA’ influence on neural stem cells’ differentiation ability. Methods The NSCs were prepared after isolation from the E14 embryonic mouse cortex and cultured in proliferation medium, the cells were then cultured in the differentiation medium with the immunostaining method to detect whether they can differentiate into neurons and astrocytes normally. Meanwhile Realtime-PCR was used to detect the expression level of several miRNAs between NSCs and the differentiated cells，miRNAs or the inhibitor were then transfected into the NSCs by the high efficiency of lipid reagent, western blot was used to detect the miRNAs or the inhibitor’s influence on the differentiation ability of NSCs. Results Establish the model of mouse neural stem cell culture platform successfully, detect that a set of miRNAs expressed differently between the NSCs and differentiated cells, Western-Blot showed that the antisense of miRNA-124 could reduce the expression of neural specific marker protein β-tubulin III obviously (P<0.01), while overexpression of miR-137,128 could upregulate β-tubulin III somehow. Conclusion the lipid reagent can transfect the miRNAs into the NSCs efficiently and miR-124,137,128 probably play an important role in the differentiation of NSCs.

Survey of health-related quality of life in population of 6 Chinese cities

Ting-fang PAN Chao-zeng SI Hui-jing HE Bin WANG Guang-liang SHAN

2011, 31(6):  636-641. 

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Objective The aim of this study is to establish normative values of the SF-36 health survey in 6 Chinese cities so that we can evaluate the quality of life in different Chinese population. Methods We scored the 8 domains of the SF-36 of the general population and different groups. The score of the general population was compared with the norms of Hong Kong and US. The stepwise regression analysis was used to calculate the effects of demographic variables on the SF-36 scale scores. Results The score on physical health is (77.54±15.96) and on mental health is (71.29±17.86). All sub-scales scores were similar trend of change between Chinese general population and Hong Kong general norm. Gender, age and living environment had effects on the SF-36 scores. Conclusion SF-36 health survey can be used to measure the impact of health statement on the quality of life. We can develop the norms of SF-36 by different demographic groups.

MicroRNA-144 Regulates Erythroid Differentiation of K562 Cells

Xiao-yan WANG Chong-liang JIANG Yong ZHU Fang WANG Jia YU Tao FENG

2011, 31(6):  642-646. 

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Objective: We aimed to study the influence of miR-144 on human erythroid differentiation via targeting the retinoblastoma protein ( RB). Methods: K562 cells were treated with hemin to differentiate into mature red cells. Real-time PCR analysis was used to detect the changes of miR-144 expression during erythroid differentiation. K562 cells were transfected with oligonucleotides (mimic-144) and the influence of erythropoiesis was measured. The target gene of miR-144 was identified by using bioinformatics analysis combined with Dual-luciferase reporter and Western blot analysis. Results: MiR-144 was significantly increased during hemin-induced K562 erythroid differentiation（P<0.05）. Over-expression of miR-144 in K562 cells promoted the stimulation of γ-globin and CD235a. Moreover, RB was identified as a direct target of miR-144. Expression of RB increased slightly and then reduced obviously in erythroid differentiation . Grey mean of RB/GAPDH minimum point: 12h 0.092±0.007（P<0.05 compared with 0h）. Conclusion: MiR-144 played a positive role in erythroid differentiation of K562 cells via targeting RB.

NR2F2 accelerates mouse MC3T3-E1 pre-osteoblast proliferation

Song WANG Xia-hui XIONG Ning ZHU Mei-Hong CHEN

2011, 31(6):  647-651. 

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Objective To study the role of NR2F2 in murine pre-osteoblast proliferation. Methods Quantitative real-time PCR was used to measure the mRNA level, MTS assay was used to assess the cell proliferation rate, flow cytometry was used to figure out the cell cycle distribution, and ELISA-Brdu was used to check the rate of DNA synthesis. Results When murine pre-osteoblast MC3T3-E1 cell proliferation was accelerated, the expression level of NR2F2 remarkably increased to 4.57±0.30 (p<0.01) fold compared with that of the control. Over-expression of NR2F2 accelerated MC3T3-E1 cell proliferation (p<0.01), and the proportion of cells in S phase markedly increased to 2 fold compared with that of the control, and the proportion of cells in G2/M phase also went up (p<0.05). More Brdu was incorporated into NR2F2 over-expressing cells, indicating the increase of DNA replication rate (p<0.01). Conclusion NR2F2 accelerates murine pre-osteoblast MC3T3-E1 proliferation through increasing the proportion of cells in S phase.

Proteomic Analysis of Differential Protein Expression in 293T Cells after Overexpression of Enterovirus 71 3C

Xiao-bo LEI Zhen-min SUN Xin-lei LIU Li-li REN Jian-wei WANG

2011, 31(6):  652-655. 

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Objective To explore the effects of 3C protease on host cell proteins by proteomics techniques. Methods 293T cells were transfected with plasmid encoding EV71 3C, cell lysates were analyzed by using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS) analysis. Down-regulated protein was confirmed by Western blot analysis. Results The expression of EV71 3C in 293T cells is confirmed. A total of 5 altered proteins were identified after 3C overexpression. Among them, expression of 4 proteins increased, while expression of 1 protein decreased. Down-regulated protein HSPA4 was confirmed by Western blot. HSPA4 expression was determined by performing densitometric analyses in HeLa cells. The results show that HSPA4 expression was 46.8%±10.1% (P＜0.05) compared to normal cells. Conclusions Overexpression of EV71 3C regulates the protein expression in 293T cells.

Suppression of cell invasion by galanin in rat pituitary adenoma cells

Ke-qian MA Jing SUN Jie LI Zhu-qing YAN Run-chun LU Wang JIA Zhi-qing XU

2011, 31(6):  656-660. 

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Objective To study the role of galanin and its receptors on cell vitality and invasion in rat pituitary adenoma cells. Methods Rat pituitary GH3 cells RNA was extracted to determine the expression levels of galanin and its receptors with RT-PCR; Matrigel-coated Transwell chamber was used to quantify invasiveness of rat pituitary GH3 cells in the presence or absence of galanin or GalR2 agonist AR-M1896. The effect of galanin or GalR2 agonist AR-M1896 on the cell vitality was determine by MTT assay. Results Expression of galanin and all three galanin receptor subtypes were detected in the rat pituitary GH3 cells and expression levels of GalR2 were particularly high. There is no significant difference in cell proliferation levels between the presence and absence of galanin or AR-M1896 at 12h, 24h, 36h. However, the cell invasion through Matrigel was decreased by approximately 49% in rat pituitary GH3 cells with galanin treatment(100nM) compared with control group (p<0.01). AR-M1986 (100 nM) also showed a suppression of cell invasion of 46% (p< 0.05) compared with control group. There were no significant difference in suppression of cell invasion between AR-M1986 and galanin groups. Conclusion Galanin inhibits cell invasion, at least partially via GalR2, in rat pituitary adenoma cells.

Hypocholesterolemic effect of total stilbene from Cajanus cajan L. on hyperlipidemic rabbit model and regulatory mechanism

Xian-rong GUO Qing-feng LUO Xiao-min KANG Hong-chao YIN JIan-yong SI

2011, 31(6):  661-666. 

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Objective To investigate the possible hypolipidemic effects of total stilbene from Cajanus cajan L.(TSC) in rabbits fed with a high-lipid diet and the mechanism of regulation of cholesterol metabolism. Methods Thirty-six male rabbits were divided into six groups: control group, high lipid model group, TSC-treated with 200、100、50 mg/kg，and Xuezhikang treated group. The effects of TSC were investigated by monitoring serum and liver lipid profile in rabbits. THP-1 cells were induced to macrophages by PMA. The macrophages were labeled with [3H]cholesterol. These cells were divided into five groups, i.e, normal group, Cajanin high concentration group (CH), Cajanin mean concentration group (CM), Cajanin low concentration group (CL) and estradiol(E2) group. The cells were cultured for 24 hours in the serum-free RPMI 1640 medium containing 2mg/ml bovine serum albumin(BSA) in the presence or absence of Cajanin (30 μmol/L, 3 μmol/L, 0.3 μmol/L) or 17β-estradiol ( 1 μmol/L) . The macrophages were washed with PBS again and incubated in the serum-free RPMI 1640 medium containing apoA-I (10μg/ml) for 12 hours. The radioactivity of samples of both supernatants and cell lysates were determined using the liquid scintillation counter (Beckman, LS 6000SC, USA). Fractional cholesterol efflux was calculated, and the ABCA1 protein was quantified by Western blot. Results In TSC(200 mg/kg) group，the serum and hepatic TC were reduced by 35.36% and 35.97%（P<0.01）；the TG contents of serum and liver were also lowered by 12.34%（P<0.05）and 41.32%（P<0.01）. At the same time, serum LDL-C decreased by 29.07%（P<0.01） as compared with high lipid model group. The cholesterol efflux percentage from macrophage was increased by 55.39% and 24.97%（P<0.01） respectively in CH and CM group. The expression of the ABCA1 was significantly increased in CH group. Conclusions TSC has potential hypolipidemic effects，possibly via up-regulating the ABCA1 protein expression, subsequently resulting in increased macrophage cholesterol efflux and RCT.

DNA vaccine encoding Ebola virus envelope glycoprotein inducing high-titer neutralization antibody in mice

Gui-gen ZHANG Jian-she LANG Hong-liang WANG Kang-tai LIU Cheng-yu JIANG

2011, 31(6):  667-671. 

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Obejective To study the characteristics of neutralization antibody in mice induced by DNA vaccine encoding Zaire ebolavirus glycoprotein (GP) and develop DNA vaccine against Ebola virus. Methods Zaire Ebolavirus GP encoding plasmid Peak13CD5LGP was constructed. 100 ?g dosage of Peak13CD5LGP was used to immunize the mice, the titer of neutralization antibody against Ebola Zaire virus glycoprotein was assayed by ELISA using Ebola Zaire virus glycoprotein subunit GP1-Fc fusion protien as capture antigen. Results After boost immunization with 100 ?g dosage Peak13CD5LGP plasmid, the titer of neutralization antibody of anti-GP was 1:2000, and both efficiency and specificity of neutralization antibody was verified by western blot. Conlusion 100?g dosage of DNA vaccine encoding Zaire Ebolavirus envelope glycoprotein is able to induce persistent and high level of neutralizing anibody, and may be potential valuable vaccine against the deadly Zaire Ebolavirus.

Effects of Ghrelin on the hippocampus neuronal apoptosis and the cognitive function of diabetic rats

Lou-yan MA Dong-min ZHANG Ying ZHANG Yuan GAO Li-yin CHAI Ke-xiang ZHAO Li XIA Qian XIAO

2011, 31(6):  672-678. 

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Objective To explore the effects of Ghrelin on the hippocampus neuronal apoptosis and the cognitive function of diabetic rats. Methods 40 SD male rats were meanly divided into normal group, diabetic modle group, Ghrelin treatment group, Ghrelin and D-lys-3-GHRP-6 treatment gooup randomly. Streptozotocin-diabetic rats model were established, diabetic rats with depression were screened by open-field test, learning and memory behaviors were measured using a spatial version of the Morris water maze test. The mRNA level of caspase-3 was examined by RT-PCR. The protein expression of capase-3, BCL-xl were examined with immunohistochemical method. Meanwhile the hippocampus neuronal apoptosis were measured by TUNEL. Result Comparing to the normal group, learning and memory level of diabetic group and Ghrelin + D-lys-3-GHRP-6 trreatment group decreased transparently（P<0.05）, the mRNA and protein level of caspase-3 in hippocampal neuron increased and the protein expression of BCL-xl were descendent (P<0.05), the number of neuronal apoptosis increased markedly（P<0.05）. Learning and memory level in the Ghrelin treatment group were much better than diabetic modle group（P<0.05）, the changes of caspase-3 in hippocampus significantly reduced（P<0.05）and the expression of BCL-xl in hippocampus inceased in the Ghrelin treatment group and the number of apoptotic neurons of the hippocampus was decreased remarkably (P<0.05).. Conclusion Ghrelin could improve cognitive ability in diabetic rats ,which increased the expression of BCL-xl and decreased the expression of caspase-3 to inhibit hippocampus neuronal apoptosis.

Acupuncture analgesia in childbirth the function and mechanism

Qing-shuang ZHU Feng-ying WANG Xiao-yan SUN

2011, 31(6):  679-682. 

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Objective Observation of labor analgesia acupuncture and action mechanism of the clinical effect, looking for a safe and effective, simple and convenient method of labor analgesia. Methods The stochastic sampling method choice acupuncture group 16 examples, the acupuncture + electricity stimulation group 29 examples and the control group 46 examples, (VRS ache staging) evaluate the analgesic effect by the visual simulation grading law, the record course of childbirth progress and the postpartum hemorrhage situation, the newborn Apgar grading and the newborn body mass, the observation acupuncture to the maternal infant the influence. (HPLC) examines around the acupuncture with the highly effective liquid phase chromatography in the peripheral blood noradrenalin (NA) and β-endorphin (β-EP) the content change, discusses this method the analgesia mechanism. Results After acupuncture analgesia, the VAS grading drops, the ache alleviation rate 82.76%, are higher than the control group obviously 17.94%, and enhances in obviously the peripheral blood β-EP the density, after the acupuncture, the blood β-EP and NA respectively be 5.20±0.93 mg/L and 1230±18ng/L, obviously is higher than the control group 4.38±0.67mg/L and 783±31ng/L(P<0.05). Conclusions The acupuncture can reduce the first course of childbirth childbirth ache effectively, its analgesic effect possibly is enables in the brain through the acupuncture to have the analgesia function to hand over the nature β-EP content to increase obviously concerns.

The activation of P38MAPK signaling pathway increases intracellular Aβ production in SH-SY5Y

Xue-wen GUO Liang LI

2011, 31(6):  683-687. 

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Objective To investigate the effect of the activation of P38MAPK signaling pathway on the intracellular β-amyloid(Aβ) production in human neuroblastoma SH-SY5Y. Methods SH-SY5Y cells were used to test intracellular Aβ levels by ELISA after activation of P38MAPK signaling pathway by anisomycin for 1 and 72 h.The mRNA and protein levels of APP, PS1 and BACE1 were examined by real time quantitative PCR and Western blot respectively. Results The expression of Phospho-P38MAPK was increased after anisomycin treatment 1and 72 h; the production of Aβ1-40 was about (46.88±3.60) pg/mL[p<0.05,Con group was (39.09±1.60) pg/mL]after anisomycin treatment for 72h; APP and PS1 mRNA was increased after anisomycin treatment for 1h, while APP, PS1 and BACE1 mRNA was significantly increased after anisomycin treatment for 72h;the protein levels of APP, PS1 and BACE1 were increased as 1.45-fold,1.38-fold and 1.60-fold respectively as control after anisomycin treatment for 72h while nothing alter for 1h. Conclusion Activation of P38MAPK signaling pathway could increase Aβ production and play a role in the pathygensis of Alzheimer's disease(AD).

Effect of Aβ1-42 on expression of T514 phosphorated CRMP2 in SH-SY5Y cells

Tian-rui ZHU Xiao-hong LI Min WANG Yuan-yuan ZHANG Dong WANG

2011, 31(6):  688-693. 

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Objective To investigate the effect of β-amyloid peptide 1-42( Aβ1-42) on neurite outgrowth, microtubule structure and expression of T514 phosphorated collapsin response mediator protein 2 (CRMP2) in SH-SY5Y cells. Methords Making cells model by adding aggregated Aβ1-42 in the culture medium of all-trans retinoic acid induced neuroblastoma cells SH-SY5Y. MTT methord was employed to identify the viability of SH-SY5Y cells. Cytochemistry was used to measure neurite length of induced SH-SY5Y cells. The intracellular microtubule structure and distribution of T514 phosphorated CRMP-2 was detected by immunofluorescence . Westen blot was used to detect the influence of Aβ1-42 on total expression of T514 phosphorated CRMP-2 . Results The viability of SH-SY5Y showed significant decrease in response to 24h aggregated Aβ1-42 exposure at 0.1 and 1μmol/L (p<0.01). Neurites measurement showed that 1μmol/L aggregated Aβ1-42 lead to obvious neutrite retraction (p<0.05). Immunofluorescence showed the CRMP2 immuno-positive expression was higher in both experimental groups (0.1 and 1μmol/L ) (p<0.01) . Westen blot showed the totle expression of T514 phosphorated CRMP-2 in both experimental groups (0.1 and 1μmol/L ) maintain significant higher level compared with control (p<0.01). Conclusion Aβ1-42 can increase intracellular expression of T514 phosphorated CRMP-2 and have a negtive effect on microtubule structure and neurite outgrowth.

PTN promotes the differentiation of human umbilical cord blood mesenchymal stem cells into dopaminergic neuron-like cells

Dan-dan XUE Hai-mei SUN Feng-qing JI De-shan ZHOU

2011, 31(6):  694-699. 

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Objective To investigate the possible mechanism of CB-MSCs differentiated into DA neuron-like cells induced by PTN. Method Preparation of the mononuclear cell fraction was performed by the Ficoll gradient technique according to the manufacturer’s instructions. rPTN was added as inducing group (rPTN group), passage 1 CB-MSCs with H-DMEM as control group (CON group). 48hours later, RT-PCR and immunofluorescence were employed to indentify the expression of PTN and its receptors SDC3、ALK, also to indentify the expression of TH、DAT and NSE. Results During the induction, cell morphology changed obviously, the expression of TH, DAT, and NSE increased .The ratio of positive cells were: TH(CON group): 3.77±0.42，（rPTN group）: 7.91±0.55；DAT(CON group): 1.89±0.28，（rPTN group）: 为6.21±0.77；NSE(CON group): 4.35±0.76，（rPTN group）: 7.67±0.97；The expression of SDC3 also increased comparing with the CON group. Our study discovered PTN can promote the PTN secretion of CB-MSCs. The ratio of positive cells were: PTN(CON group): 14.38±0.54，（rPTN group）: 20.21±1.51；SDC3(CON group): 5.70±0.78，（rPTN group）: 10.02±1.45；ALK(CON group): 4.87±0.34，（rPTN group）:5.01±0.99; Conclusion PTN could induce CB-MSCs differentiate into DA neuron-like cells by prompting CB-MSCs express PTN and its receptor SDC3.

Depressive disorder increase expression of NF-κB in rat vessel tissue

Ying CHEN Zhi-wen CHANG Lin YANG Yan-ling ZHAI Yong ZHANG Wen-juan LIN

2011, 31(6):  700-703. 

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Objective To investigate the relationship between depressive disorder and inflammation markers in depressive rats induced by Forced Swimming Test (FST). Methods SD rats were randomly divided into controlling groups and modelling groups, Depression animal model was reproduced with FST in 21days. Modeling group included SS group (FST rats with 0.9% sodium cloride treated) and SI group (FST rats with Imipramine treated). all peripheral blood serum samples were respectively measured using enzyme linked immunoassay (ELISA) technique for concentration of MCP-1 and Immunoturbidimetric Assay for hsCRP. The expression of MCP-1 and NF-κB p65 in aorta tissue were observed by immunohistochemistry. Results There were high level of concentration of hsCRP and MCP-1 in peripheral blood serum samples and increased expression of MCP-1 and NF-κB p65 in aorta tissue of SS groups (P<0.05). SI groups had decreased the expression of inflammation markers in blood and aorta tissue. Conclusions Depressive disorder activate the inflammatory reaction with increased expression of NF-κB.

3DUS Imaging in Assessment of Gastric Accommodation in Healthy Subjects

Jing ZHANG Zhong-hui XU Mei-yun KE Zhi-feng WANG Xiao-hong SUN Wei ZHAO Qing DAI

2011, 31(6):  704-708. 

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Objective To evaluate gastric accommodation using 3D ultrasound imaging and Perfusion Nutrient Load test（P-NLT） by means of gastric volumes calculation and to verify validation of P-NLT in evaluation of gastric accommodation in healthy subjects. Methods 10 healthy subjects received P-NLT through a nasal-gastric tube with a constant rate of 50 mL/min (0.75 kcal/mL). Meanwhile 3DUS and 2DUS were used for imaging of PGV and PGA in different day, respectively. During the perfusion, visual analogue scale (VAS, 0～10) was used to evaluate satiety .The nutrient volume in different satiety scale and time consume was recorded during P-NLT. Then the measure values in 3DUS were compared with corresponding parameters in 2DUS. Results No difference was observed in the amount of nutrient liquid at minimal satiety and maximal satiety between the first and the second P-NLT study. PGV (p<0.01) and PGA (p<0.05) at maximal satiety measured by 3DUSwere both significantly correlated with MV. Furthermore, PGV at minimal satiety measured by 3DUS (p<0.05) was significantly correlated with TV. Significant differences were found in PGV (p<0.05) and PGA (p<0.01) at maximal satiety between 3DUS and 2DUS, and there was significant difference in PGA (p<0.01)at minimal satiety in two methods, too. But the curve estimated between MV and PGA or PGV at maximal satiety used by 2DUS was parallel with that in 3DUS. Conclusions 3DUS is superior to 2DUS for its accuracy and precision in volume estimation and more available for proximal gastric accommodation. P-NLT is a feasible and repeatable method in assessment of proximal gastric accommodation.

Novel non-viral vector O-CHCS as gene vector for transfecting HBD2 to L929 cells and biological activity detection of expression products

Nan CUI Xiao-nan FANG Ni ZHANG Ming-mao CHEN Xue-min LI Xin-nian CHEN

2011, 31(6):  709-712. 

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Objective To explore O-CHCS’s feasibility of being a gene vector for transfecting L929 cells. Methods We constructed plasmid pCMV- hBD2, and the plasmid was introduced into L929 cells by O-CHCS. Total RNA were extracted from the cultured cells, RT-PCR were performed with specific primers for HBD2 and RT-PCR amplification products were identified with agarose gel electrophoresis;the expression of HBD2 protein was verified by Western blot;the antibacterial activity of purpose protein was defected by Kirby-Bauer disk diffusion method, at the same time put the Lipofect as the positive control. Results RT-PCR amplification products by agarose gel electrophoresis were the same size as the target gene by observation; HBD2 gene expression at the protein level was detected by Western blot ；mediums from the cultured cells transfecting with the pCMV-hBD2 could format antibacterial circle against the Staphylococcus aureus, and these results were as same as the Lipofect. Conclusion The eukaryotic expression vector pCMV-hBD2 was successfully constructed and was introduced to the L929 cells, by doing this we verified O-CHCS could be used as a gene vector for HBD2 , and provided a economic and convenient way to artificial synthesis of HBD2.

Small Continuous Curvilinear Capsulorhexis during Phacoemulsification in Senile Intumescent Cataracts

Xiao-wei LIU Zheng WANG Wei-hong YU Zao-wen WANG Jin MAO

2011, 31(6):  713-716. 

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Objective To evaluate the safety of small-capsulorhexis skills during phacoemulsification in senile intumescent cataract patients and the short-term surgery outcomes. Methords 143 senile intumescent cataract eyes (143 patients) were included in this study. They received phacoemulsification surgeries with a deliberately small continuous curvilinear capsulorhexis(CCC) (about 4mm) that was secondarily enlarged to about 6mm after intraocular lens had been inplanted. The rate of CCC completed and intraoperative and postoperative complications and short-term surgical outcomes had been statistically analyzed. Results CCC had been completed in 132(92.3%) intumescent cataract patients and secondly CCC was needed in 82 (57.3%) of them. Four failed cases suffered from breaking of the capsule and vitreous loss. Conculsion: Small continuous curvilinear capsulorhexis could help prevent unexpected radial tears of the capsulotomy during phacoemulsification in intumescent cataract patients.

Effects of dexamethasone on two acute liver injury models

Xiao-rong PENG Hong-xu LU Juan LIU Fei HE Gui-ju LI Jun XIE Li CHEN Li ZHANG

2011, 31(6):  717-718. 

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Cultivation and identification of human benign biliary duct cicatrix fibroblasts

Hao ZOU Tao WANG Xiao-wen ZHANG Hong ZHU Zhao NA Xu SHEN

2011, 31(6):  719-720. 

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CFSE/PI double labeling detect cytotoxicity of CIK to lung cancer cell line A549pretreated by cisplatin

Ben-ling XU Quan-li GAO Long YUAN Xu-hua ZHANG Rui-hua FAN Xue LIU Jin-dong GUO

2011, 31(6):  721-722. 

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