Abstract:Objective To construct the conditionally replicative adenovirus vector pAd-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55Kcarrying early growth response gene-1（Egr1） promoter and tumor necrosis factor related apoptosisinducing ligand (TRAIL) gene,and to observe the effects of the vector combined with 2 Gy irradiationon the TRAIL expression in MDA-MB-231 cells.Methods Egr-1 promotor sequence was cloned from pMD18 T-Egr1,TRAIL was constructed the downstream of Egr1 promoter,pShuttle-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55K (CRAd.pEgr1-TRAIL) was constructed,after the adenovirus vector was packaged successfully,MDA-MB-231 cells were infected with them and irradiated with X-rays.Real time PCR method and ELISA wereused to detect the expression levels of TRAIL mRNA and protein,respectively.Six groups in theexperiment were set up: control,2 Gy,CRAd.p,CRAd.pEgr1-TRAIL,CRAd.p + 2 Gy and CRAd.pEgr1-TRAIL + 2Gy. Results The recombinant adenovirus vector pAd-Egr1-TRAIL-hTERT-E1A-E1Bp-E1B55K was constructedand packaged successfully. The expression level of TRAIL mRNA in MDA-MB-231 cells transfected withthe vector of 5 MOI for 24 h following 2.0 Gy X-rays irradiation began to increase and arrived tothe top 8 h later in various groups,then declined.The expression level of TRAIL protein in MDA-MB-231 cells began to increase 6 h after irradiation and reached to the peak 24 h later,then declined48 h later.There were significant differences in the expression levels of TRAIL protein betweenCRAd.pEgr1-TRAIL + 2.0 Gy and other groups at the same time point (P<0.01).Conclusion Therecombinant adenovirus vector is obtained successfully,and the TRAIL mRNA and protein expressionlevels in MDA-MB-231 cells can be increased significantly by the vector combined with 2.0 Gy X-raysirradiation.

Abstract:Objective
 To investigate the influence of salvianolic acid B on the expressions of TGF-β1/Smad signalingpathway related proteins in human lung fibroblasts,and to explore the mechanism of the inhibitoryeffect of Sal B on TGF-β1-induced lung fibroblast activation.Methods The human embryonic lungfibroblasts (MRC-5) were cultured in vitro and randomly divided into control group ( the cells were cultured with DMEM without TGF-β1 or Sal B),Sal B group (the cells were cultured with 10  μmol•L^-1 Sal B),TGF-β1 group (the cells were cultured with 10 μg•L^-1 TGF-β1),and TGF-β1(10 μg•L^-1) + SalB (10  μmol•L^-1) group.The protein levels of p-Smad2,p-Smad3,TβRⅠ, and Smad7 in the fibroblastsin various groups were detected by Western blotting method. Results Compared with control group,the expression levels of p-Smad2,p-Smad3, and TβRⅠ proteins in TGF-β1 group were significantly increased (P<0.05),and the expression level of Smad7 protein was decreased (P<0.05).Compared with control group,the expression levels of p-Smad2,p-Smad3,TβRⅠ, and Smad7 proteins in lung fibroblasts in Sal B group had no significant change (P>0.05).Compared with TGF-β1 group,the expression levels of p-Smad2,p-Smad3, and TβRⅠ in TGF-β1+ Sal B group were descended (P<0.05),and the expression level of Smad7 was increased (P<0.05).Conclusion Sal B could suppress the TGF-β/Smad signaling pathway in lung fibroblasts and  to inhibit the  TGF-β1-induced lung fibroblast activation.

Abstract:Objective To explore the expression of caspase-3 in skeletal muscle of the mice in the state of cancer,and to elucidate the relationship between Caspase-3 and apoptosis,consumption of skeletal muscle protein in cancer cachexia.Methods 48 male BALB/c mice were randomly divided into cancer cachexia group and control group （n=24）.The mice in cancer cachexia  group were inoculated with  mouse  colon 26 adenocarcinoma cells.The body weights of the mice in two groups were detected daily.Eight mice in each group were executed to test the weight of left gastrocnemius,fiber crosscut area,the expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),Caspase-3 proteins and the apoptotic rate of gastrocnemius cells on day 8,14, and 20, respectively.Results The mice in cancer cachexia  group appeared cachectic symptoms on day 14,the non-tumor body weight was decreased more than 20% of that in control group（P<0.05）.Compared with control  group at the same time,the mouse body weight,non-tumor body weight,the weight of left  strocnemius and the fiber crosscut area of the mice  in cancer cachexia  group were obviously decreased with the prolongation of inoculation time （P<0.05）,whereas the expression levels of TNF-α,IL-6,Caspase-3 proteins and the apoptotic rate of muscle cells were obviously increased after tumor inoculation（P<0.05）.The level of Caspase-3 protein was negatively correlated with the weight of gastrocnemius and fiber crosscut area r=0.716,P<0.05;r=－0.694,P<0.05）,and the level of Caspase-3 was positively correlated with the levels of TNF-α and IL-6（r=0.742,P<0.05;r=0.675,P<0.05）.Conclusion Caspase-3 may be a key factor in the protein comsumption of skeletal muscle in cancer cachexia.

Abstract:Objective
To investigate the influence  of arsenic trioxide(AS2O3) in the vasculogenic mimicry(VM ) of HepG2 cells，and to preliminary clarify the possible mechanism of inhibition of AS2O3 on the VM.Methods The mean inhibitory concentration (IC50)  of AS2O3  72 h after treatment of HepG2 cells was calculated by CCK-8 assay.The HepG2 cells were cultured on 3-D Matrigel and randomly divided into control group,1/2 IC50AS2O3 group and IC50 AS2O3 group.IPP software was used to calculate the number,length and area of  VM,and the expression levels of  VM-related proteins VE-cadherin and MMP-2,apoptotic-related protein caspase-3 and proliferation-related protein PCNA were detected by Western blotting method.Results  The IC50 of AS2O3 was 10 μmol•L^-1 72 h after reatment of HepG2 cells.The number,length and area of VM in 1/2 IC50 and IC50AS2O3 groups were significantly lower than those in control group (P<0.01); the number,length and area of VM in IC50AS2O3 group were also lower than those in 1/2 IC50AS2O3 group (P<0.05).Compared with control group,the expression levels of VE-cadherin and MMP-2 in 1/2 IC50 and IC50 AS2O3 groups were decre ased (P<0.05), and  the expression levels of caspase-3 and PCNA had no significant change (P>0.05).Conclusion AS2O3 can inhibit the forming of VM of HepG2 cells,which indicated that its  mechanism may be related to  inhibiting the expressions of VE-cadherin and MMP-2.

Abstract:Objective
To study the effects of benzyl propionate nandrolone (BPN) on the nicotinamide phosphoribosyl transferase (Nampt),insulin receptor substeate-2(IRS-2) and pancreatic duodenal homeobox-1(PDX-1)expressions,cell cycle changes as well as insulin secretion in pancreatic islet cell NIT-1 lines,and to explore the influence of BPN in the Nampte xpression in NIT-1 cells and insulin signaling molecules in high glucose oxidation stress.Methods The NIT-1 cells were cultured with different concentrations  (5.6,11.1,16.7, and 27.6 mmol•L^-1) of glucose,then they were treated with 10 mg•L^-1 BPN for 48 h with no BPN treatment as corresponding control groups. The expression levels  of Nampt,IRS-2, and PDX-1  were tested by Western blotting assay. The changes of cell cycle were determined by FCM and the cell insulin secretion levels were measured with radioimmunoassay. Results Compared with corresponding control groups,the expression levels of Nampt,IRS-2, and PDX-1 proteins in the NIT-1 cells in various BPM groups were increased (P<0.05 or P<0.01).The G0/G1 phase arrest was relieved (P<0.01) when the cells was cultured in low glucose(5.6 mmol•L^-1) co ndition,and the G2/M block was remitted significantly in  high glucose  (27.6 mmol•L^-1) condition (P<0.01),furthermore,the cell insulin secretion was promoted compared with control groups except 11.1 mmol•L^-1 glucose group(P<0.01).Conclusion BPN can promote the expression levels of  Nampt,ISR-2 and PDX-1 proteins  in NIT-1 cells.There is close relationship between the Nampt expression in NIT-1 cells and insulin signaling pathway and BPN prevents the cells from insulin resistance.

Abstract:Objective
To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP),and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells.Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells.Different concentrations (0.5  and 1.0 mmol•L^-1) of  isopropyl-β-D-thiogalactopyranoside (IPTG) and cell culture temperatures (22℃ and 37℃) were used to optimize the protein expression. The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used toidentify the Tat-GFP protein,and confocal laser scanning microscope (CLSM) was used to examine the cell penetration of Tat-GFP protein.Results There was no significant difference in the Tat-GFP protein production induced by 0.5  and 1.0 mmol•L^-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize  the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The  Tat-GFP protein highly expresses  in the supenatant of Escherichia coli i BL21 cells   at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.

Abstract:Objective To investigate the biofilm(BF) formation rule of nontypeable Haemophilusinfluenzae (NTHi) in vitro,and to observe the internal structure of BF by scanning electron microscope (SEM).Methods NTHi ATCC49247 was investigated in the present study，Pseudomonas aeruginosa (PA) PAO1 was cultured as positive control,at the same time blank control group was set up.The BF of the  bacteria were cultured and then collected on day 1,2,3,4,5,6, and 7.The BF formation was detected by crystal violet staining and plate counting and the structure of BF formed by ATCC49247 was observed under SEM on day 3.Results The plate colony counting of biofilm BF  by ATCC49247 and PAO1 raised during first 3 d,and then declined to (0.823 6±0.007 5)×10⁷ cfu•mL^-1 and(0.942 6±0.019 9)×10⁷cfu•mL^-1 respectively on day 7. The differences between two groups were statistically significant on day 3,4,5, and 6(P<0.05).The differences between different time points in the same bacteria group were statistically significant(P<0.05).The densities of  BF formed by ATCC49247 and PAO1 raised during the first 3 d.The absorbances on 570 nm wavelength(A570) in two groups were 2.717 4±0.017 2 and 2.885 3±0.039 0 ,respectively;and then the A570 values in two  groups declined to 0.151 7±0.074 5 and 1.196 9±1.108 5, respectively on day 7;the differences between bacteria groups and blank control were statistically significant(P<0.05);the differences between two bacteria  groups were  statistically significant on day 3,4,5, and 6(P<0.05);the differences between different time points in the same bacteria group were statistically significant (P<0.05).On day 3,the obvious BF formed by ATCC49247 were observed under SEM.Conclusion BF could be formed by NTHi in vitro；crystal violet staining,plate colony counting and SEM could be taken as conventional methods to detect BF.

Abstract:Objective To investigate the  influence of  high-fat diet in the intestinal flora，fecal weight and its water content in rats，and to clarify the effect and significance  of high-fat diet in the occurrence of obesity forming.Methods Twenty Sprague-Dawley(SD)  rats were randomly divided into normal diet（ND）group and high-fat diet（HF）group（n=10）.The rats in ND group were fed with normal diet, and the rats in HF group were fed with diet rich in oil and fat.The fresh feces were collected separately on days 1，15，30, and 49 for analysis of weight,water content，and intestinal flora.Results On the 49th day，the wet weight and water content of feces of the rats in ND group were (6.61±0.17）g  and（37.07±3.04）%, respectively,while those in HF group were（4.46±0.30）g  and （18.04±2.23）%(P<0.05).Compared with ND group ，the fecal pellets in HF group were increased obviously from the 7th day(P<0.05).There were obvious changes in intestinal microbial populations of HF group.The counts of enterococci， bifidobacteria， Lactobacillus，Bacteroides bacteria were significantly decreased on the day 49，but the count of Escherichia coli  was increased significantly（P<0.05）.Conclusion High-fat diet can result in decrease for weight，water content,and  pellets of feces;it can change the structure of intestinal flora.As result,there is a possibility that all ofchanges above can promote obesity in the future.

Abstract:Objective
To study the expressions of nicotinamide phosphoribosyl transferase (Nampt) in main energy metabolism organs (liver,pancreas,skeletal muscle,and kidney) of the rats with type 2 diabetes mellitus (T2DM)， and to explore the correlation between the expression and distribution of Nampt and the occurrence of diabetes.Methods The SD rat diabetes model was established by injecting with streptozotcin(STZ).The SD rats were randomly divided into diabetes group and control group.Immunohistochemical Envision staining assay was used to detect the distribution and protein expressions of Nampt in liver,pancreas,muscle, and kidney tissues of the rats,at the same time the blood glucose and serum insulin levels were also be detected. Results The blood glucose level of the rats in diabetes  group was significantly higher than that in control group (P<0.01),and the fasting insulin level was lower than that in control group (P<0.01).The Nampt expression in the liver tissue of the rats in diabets group was significantly increased,which distributed near the hepatic sinus in diabetes rats,and the Nampt expression was also increased in skeletal muscle in which the whole cell  was thick dying;the Nampt expression  mainly distributed in the renal tubular epithelial cells.Compared with control group,the positive expression rates of Nampt  in liver,skeletal muscle, and kidney tissues of the rats in diabets group were significantly increased (P<0.05 or P<0.01). There was nearly no Nampt expression in pancreas tissue of the rats in diabetes group and the Nampt expression level was significantly lower than that in control  group (P<0.01).Conclusion The Nampt expressions are much different in main energy metabolic organs of the rats with diabetes.It issuggested that Nampt may be used as a specific indicative marker in the process of diabetes.

Abstract:Objective
To detect the expression of ARK5 in hepatocellular carcinoma(HCC) tissue and  hepatoma SMMC-7721 cells,and to investigate its effect on the growth of hepatoma cells.Methods The expression levels of ARK5 mRNA and protein were determined by RT-PCR and Western blotting in 30 cases of HCC tissue,paracarcinoma tissue,SMMC-7721 cells, and hepatic cells LO2.The SiRNA of ARK5 and negative control (NC) siRNA were constructed and transfected into the  SMMC-7721 cells,and used as experimental group and negative control group;at the same time blank control group was set up.The proliferation activity and apoptotic rate of transfected cells were detected by MTT assay and flow cytometry (FCM). Results The PCR and Western blotting results showed that the expression levels of ARK5 mRNA and protein in HCC tissue and SMMC-7721 cells were significantly higher than those in paracarcinoma tissue and LO2 cells(P＜0.05).The MTT assay results demonstrated that the inhibitory rates of growth of  transfected cells in experimental group at 24,48 and 72 h were (19.39±5.42)%,(23.19±0.53)%,and (20.74±1.23)%;there were significant differences compared with blank control group and negative control group (P＜0.01). The FCM results indicated that the apoptotic rate of the transfected cells in experimental group was (15.017±0.945)%,there were significant  ifferences compared with blank control group (8.770%±0.656)% and negative control group (8.763%±1.201%)(P＜0.05).Conclusion The ARK5 expression level is significantly increased in HCC tissue and hepatoma SMMC-7721 cells;the inhibition of ARK5 expression could suppress the growth of hepatoma cells and induce apoptosis.So ARK5 maybe act as a cancer-promoting gene and induce hepatocellular carcinogenesis.

Abstract:Objective To study the antitumor effect of new photodynamic therapy (PDT) applying TMPyP4 combined with nucleolin silence on cervical cancer SiHa cells in vitro,and to explore an available combination treatment project for cervical cancer.Methods The SiHa Cells were divided into blank control group,RNAi group,PDT group and PDT-RNAi group.The proliferation activities of SiHa cells were assessed by Cell Counting Kit-8 (CCK8) assay.The apoptotic rates were measured by flow cytometry (FCM) with Annexin Ⅴ/PI staining.The invasiveness abilities were assessed by Transwell assay.Results Compared with blank control group,the inhibitory rates of growth of SiHa cells in RNAi group,PDT group and PDT-RNAi group were increased significantly (P<0.01);the inhibitory rates of growth of SiHa cells in PDT-RNAi group were higher than those in PDT group and RNAi group(P<0.05).The Q value was 1.27.Compared with blank contr ol group,the apoptotic rates of SiHa cells in experiment groups were increased(P<0.05),and the  late apoptotic rate in PDT-RNAi group was also increased;there were significant differences of the apoptotic rates between PDT-RNAi group and RNAi group,PDT group(P<0.05).Compared with blank control group,the cell invasiveness abilities of SiHa cells in experiment groups were decreased;there were significant differences of the invasiveness abilities of SiHa cells between PDT group and RNAi group (P<0.05).Conclusion New PDT shows a strong photodynamic effect on the SiHa cells,which can inhibit the proliferation and invasiveness and induce the apoptosis of SiHa cells in vitro;nucleolin silence shows a good synergy effect to PDT.

Abstract:Objective
To investigate the influence of  tissue-specific growth hormone receptor(GHR) deficiency   in  type 1 diabetes in the mice at  the gene level using pancreatic β cells   combined with streptozotocin (STZ)-induced type 1 diabetes model.Methods The experiment was  divided into four groups:knockout mice group(LLc knockout group),using the homozygotes(LLc:LL+Cre)producted by  pancreatic β cell- specific expressed  recombinant enzyme  mice(RIP-Cre) and Cre-LoxP system modified GHR  mice (Floxed,LL);LL control group,containing Floxed GHR allele homozygous mice(LL);LLc STZ group and LL STZ group(STZ was used for inducing type 1 diabetes model mice).The mice with feeding glucose≥25 mmol•L^-1 were considered to be successful models.The Glucose Tolerance Test (GTT), pancreas tissue HE staining and immunohistochemistry were performed in the mice.Results The blood glucose of the mice in LL STZ group and LLc STZ group and LLc STZ group were increased after injection of STZ and the models achieved the diagnostic criteria for diabetes 16 d later.The results of GTT showed that compared with LLc control group and LLc knockout group,the blood glucose levels of the mice in LL STZ and LLc STZ groups were increased (P<0.05).There was no significant change of morphology and structure  of islets between LL control group and LLc knockout group detected by HE staining.The immunohistochemistry results showed that the  insulin level of the mice  in LL STZ group was significantly reduced compared with LL control group;the insulin level of the mice in LLc STZ group was reduced compared  with  LLc control group.Conclusion  Pancreatic β cell GHR gene knockout has no  effect on the  blood glucose and the function of β cells in the mice with STZ-induced type 1 diabetes.

目的：应用反义寡核苷酸（ASODN）技术靶向抑制生存素（survivin）基因，研究其对肝癌 SMMC-7721细胞凋亡的影响，阐明survivin反义寡核苷酸（survivin-ASODN）促进SMMC-7721细胞凋亡的作用机制。方法：设计合成survivin-ASODN序列（FAM荧光素标记）。利用脂质体介导法分别用浓度为100、200、300、400和600 nmol?L-1 survivin-ASODN转染SMMC-7721细胞(ASODN转染组)，并设空白对照组、空脂质体对照组 和正义寡核苷酸（SODN）对照组。应用流式细胞术(FCM)检测不同浓度survivin-ASODN体外转染SMMC-7721细胞24、48和72 h后SMMC-7721细胞凋亡率及细胞周期；Western blotting法检测survivin蛋白表达水平。结果：与各对照组比较，荧光素标记的ASODN转染SMMC-7721细胞24 h后，细胞生长开始受到抑制，细胞凋亡率增加，呈现剂量-时间依赖性（P<0.05）；作用48 h后，ASODN转染组细胞在G1期前出现明显的亚二倍体凋亡峰，G0/G1期细胞明显减少（P<0.05），G2/M期细胞显著增加（P<0.05），细胞被阻滞于G2/M期。与各对照组比较，不同浓度ASODN转染组SMMC-7721细胞survivin蛋白表达水平下降（P<0.05），且随作用时间的延长而降低（P< 0.05）。结论：survivin-ASODN通过抑制survivin蛋白表达、改变细胞周期进程等机制促进SMMC-7721细胞凋亡，并且呈现时间-剂量依赖性。

Abstract:Objective  To observe the effect of Mongolian medicine Tonglaga-5 Pill on the osteoporosis induced by retinoic acid in the rats,and to provide experminted basis for its clinical application.Methods 60 female Wistar rats were randomly divided into blank group,model group,Ossptide Tablets group,Tonglaga-5 Pill groups with low,medium and high doses (n=10).The osteoporosis models were induced by retinoic acid in the rats.The levels of serum calcium (Ca),phosphorus (P),alkaline phosphatase (ALP),transforming growth factor β1 (TGF-β1),insulin like growth factor (IGF-1) and bone indexes were detected and the contents of Ca and P in ash were also detected,and the changes of bone histomorphometry of the rats were observed under light microscope by HE staining.Results Compared with model group,the serum calcium and   P contents of the rats in Tonglaga-5 Pill groups were increased,but there were no significant differences(P>0.05);the ALP levels were  decreased significantly(P<0.05);the TGF-β1 and IGF-1 had no significant changes(P>0.05);the contents of calcium and P in  ash were increased in different degrees,but there were no significant differences (P>0.05).Compared with control group,the femur maximum width index and minmum width index of the rats in model group were significantly decreased(P<0.05 or P<0.01)；the femur length index was increased(P<0.05).Compared with model group,the femur maximum width index and minmum width index of the rats in Tonglaga-5 Pill groups had no changes(P>0.05).Under light microscope,the rich and full bone trabecula was found in the rat femur in control group;the bone trabecula in model group was decreased,and the bone trabecula in Tonglaga-5 Pill groups was increased.Compared with model group,the trabecular bone areas in Tonglage-5 pill groups and Ossotide group were increased(P<0.05).Conclusion  Tonglaga-5 Pill  displays a certain preventive effect on  the rats with retinoic acid-induced osteoporosis,which can blocks the bone mineral loss,bone tissue structure changes and bone quality decreasing   effectively.

Abstract:Objective To study the influence of Rhizoma typhonii extract on the growth of glioma SHG-44 cells cultured in vitro,and to explore the mechanism of the  inhibitory effect of Rhizoma typhonii extract on the growth of glioma cells.Methods The SHG-44 cells were cultured and divided into blank control group and 8,40,200,1 000  μg/L Rhizoma typhonii extract groups.The inhibitory effect of Rhizoma typhonii extract on the growth of glioma SHG-44 cells was measured by MTT assay.The secretion levels of Bax and caspase-3 proteins were examined by ELISA assay.The expression level of caspase-3 protein was examined by Western blotting method.Results Compared with blank control group,the inhibitory rates of the growth of SHG-44 cells in 200,1 000 μg?L-1 Rhizoma typhonii extract groups at 24　h,and 8,40,200, and 1 000 μg/L Rhizoma typhonii extract groups at 48 h were significantly increased (P<0.05 or P<0.01).The secretion levels of Bax and caspase-3 proteins in 40,200, and 1 000 μg/L Rhizoma typhonii extract groups at 48 h were increased compared with blank control group (P<0.05 or P<0.01).Compared with blank control group,the expression levels of caspase-3 protein in different doses of  Rhizoma typhonii  extract groups were increased significantly(P<0.01).Conclusion Rhizoma Typhonii extract can inhibit the growth of cells through up-regulating the expression of Bax protein,increasing the expression level of caspase-3 protein and activating apoptosis pathway.

Abstract:Objective To explore the changes of PH values of N,[KG-3]O-CMC/β-TCP compositive materials with different mass fractions in simulated body fluids (SBF) and their influence in the growth of MG63 cells,and to illustrate their mechanisms,and to provide reference for the further research on the bone repair materials.Methods The N,[KG-3]O-CMC/β-TCP with mass fractions of 2/1,1/1 and 1/2 were used as experimental groups,and the collagen nano calcium phosphate bone repair material as control group.The materials with  different mass fractions  were immersed in SBF and the pH values were measured by  pH meter after soaking for 7,14,21 and 28d,respectively.The MG63 cells with the concentration of 1×105mL-1 were inoculated and co-cultured in experimental and control groups,the adhesion and morphological changes of MG63 cells in each group were observed by scanning electron microscope and the cell proliferation was detected by MTT method after co-culturing for 2,4 and 6 d.Results The pH values were 6.70－7.25 in N,[KG-3]O-CMC/β-TCP (1/2) group and N,[KG-3]O-CMC/β-TCP(2/1) groups   and the pH value  in 　N,[KG-3]O-CMC/β-TCP (1/1) group was basically 7.15. The cells in N,O-CMC/β-TCP  (2/1) group formed owe full,spreading face small and less secretion,but the cells in N,[KG-3]O-CMC/β-TCP 1/2 and 1/1 groups formed in full,pseudopodia interconnection,widely spreading and more secretions under electron microscope. The proliferation rate of the cells in N,[KG-3]O-CMC/β-TCP with (1/1) and N,[KG-3]O-CMC/β-TCP(2/1) groups had no statistical differen ce compared with control group (P>0.05),but there was significant difference between control group and N,[KG-3]O-CMC/β-TCP (1/2) group (P<0.05). Conclusion The changes of pH values of N,[KG-3]O-CMC/β-TCP materials  with different mass fractions in SBF are small and the pH values are neutral;the order of the  mass fraction of  N,[KG-3]O-CMC/β-TCP to promote the growth of MG63 cells is 1/1,2/1,and 1/2.

Abstract:Objective To investigate the  effect of C-erbB-2 shRNA on chemosensitivity of mouse lung adenocarcinoma cells and its mechanism,and to find new therapy method for non-small cell lung cancer,especial lung adenocarcinoma.Methods The mouse lung adenocarcinoma Lewis cells were cultured regularly and divided into non-transfected group,pGPU6/RFP/Neo-shNC group and pGPU6/RFP/Neo-erbB-2 group.The plasmids were synthesized and transfected into Lewis cells in each group by Lipofectamine 2000.The expression levels of C-erbB-2 mRNA and protein in the cel ls in various groups were tested by RT-PCR and Western blotting method,respectively.The Lewis cells were divided into non- transfected group,pGPU6/RFP/Neo-shNC group,carboplatin group,pGPU6/RFP/Neo-erbB-2 group,pGPU6/RFP/Neo-shNC+carboplatin group and pGPU6/RFP/Neo-erbB-2+carboplatin group.The apoptotic rates of the cells in each group were detected by flow cytometry;the expression levels  of Bcl-2 and Bax proteins in each group were determined by Western blotting method.Results The expression levels of C-erbB-2 mRNA and protein in pGPU6/RFP/Neo-erbB-2 group were lower than those in non- transfected group and pGPU6/RFP/Neo-shNC.The apoptotic rate of the cells in pGPU6/RFP/Neo-erbB-2+carboplatin group was the highest in all of the groups (P<0.01);compared with the others,the expression of Bax protein in pGPU6/RFP/Neo-erbB-2+carboplatin group was increased,while the expression level  of Bcl-2 protein was decreased.Conclusion C-erbB-2 shRNA can increase the Lewis cells’ sensitivity to carboplatin.The mechanism may be that it can enhance the Lewis cells’ apoptosis induced by carboplatin through increasing the expression of Bax protein and decreasing the expression of Bcl-2 protein.

Abstract:Objective To investigate the anti-apoptosis effect of transgiutaminase 2(TG2) in osteosarcoma cell line MG-63 and to explore the mechanism of inhibiting apoptosis of tumor cells. Methods The TG2-tgrgeted siRNA was designed,and the hypoxia culture model of MG-63 cells  was established by a hypoxia incubator and the cells were divided into four groups: normal oxygen group,the  cells were cultured under normal oxygen;hypoxia group,the  cells were cultured in hypoxic incubators;control siRNA hypoxia group, the cells were cultured in hypoxic incubators after transfection of control siRNA; TG2 siRNA hypoxia group ,the cells were cultured in hypoxic incubators after transfection of TG2 siRNA.The expressions of Bax and cytochrome C(Cyt C) and the apoptotic rates were observed at different  time  (6,12,24,48,and 72 h) after hypoxia culture.Microtiter plate assay was performed to monitor the intracellular TG2 activity.RT-PCR was used to detect the mRNA expressions of TG2 and Bax.The expressions levels of TG2,Bax and Cyt C were observed by immunohistochemical staining and Western blotting method.The  apoptotic rates were analyzed using flow cytometry.Results Compared with normal oxygen group,the activity of TG2,the mRNA and protein expression levels of TG2 in hypoxia group and control siRNA hypoxia group were increased remarkably with the prolongation of  the hypoxia time (P<0.01);the expression level of Bax  protein  was decreased significantly(P<0.01),but the  expression level of Bax mRNA  had no significant change(P>0.05);the expression  levels of Cyt C protein   in cytoplasm and mitochondria and the apoptotic rates had no  markedly changes(P>0.05).Compared with hypoxia group and control siRNA hypoxia group,the expression levels of mRNA and protein of TG2 in TG2 siRNA hypoxia group were decreased significantly at different time points(P<0.01);the protein expression levels of Bax and Cyt C in cytoplasm and the apoptotic rates were increased markedly(P<0.01);the expression level of Cyt C in mitochondria was decreased(P<0.01).Conclusion TG2 can inhibit the apoptosis of tumor cells through down-regulating the  Bax expression and preventing Cyt C releasing into the cytoplasm.

Abstract:Objective To analyze the stress state of magnetic attachment in mandibular complete overdenture supported by nature roots and implant,and to provide neference for designing of clinical prosthodentics.Methods Three-dimensional finite models (model,Ⅱ,Ⅲ) of three groups of mandibular overdentures depending on the different location of the implant and natural tooth root，placing three pairs of magnetic attachment,were constructed by application of CT scanning,computer photo processing system,and Solidworks　finite element model building   software.The natural teeth and implant neck bone stress of the models in three groups under different stress were calculated and compared.Results The stress of the bone around the implant in the area of molar teeth in model with both sides of the implants under oblique load (Model Ⅱ)  was significantly increased compared with the model with one side of the implant(Model Ⅲ).The supporting bone stress of oblique load was increased compared with the vertical load;among them under the oblique load at one side’s molar teeth,the stress of the bone around the implant in the area of molar teeth in modelⅠ,modelⅡ and model Ⅲ was increased about 30％,43%,and 55%.Conclusion When there only one nature teeth remain,two implants should be at least added,one in the area of opposite cuspid,and the other in the area of  molar teeth of the same side.The magnetic attachment is better than other kinds of attachment in preventing the damage of abutment when it suffers inclined load.

Abstract:Objective To isolate and culture mesenchymal stem cells (MSCs) from human term umbilical cord and placenta， and to study the proliferative and adipogenic capacities of human fetus tissue MSCs (FT-MSCs),umbilical cord MSCs (UC-MSCs) and placenta MSCs (PL-MSCs),and to provide an experimental basis for its clinical use.
Methods The human term umbilical cord and placenta were obtained in sterile condition.The UC-MSCs and PL-MSCs were isolated by digestion using collagenase Ⅰ,the FT-MSCs were provided by Zhongke Bio-engineering Co.,Ltd.The  3th or 4th passages of MSCs were selected in this study.The cell morphology was observed by inverted microscope.The proliferative ability was detected by living cell number counting method.The cell cycle and the positive expression rates of the surface markers of three kinds of MSCs were detected by flow cytometry (FCM).The 3th generation of UC-MSCs,PL-MSCs,and FT-MSCs were induced to differentiate into adipogenic cells.18 d after being induced,the differentiated cells were stained with Oil red O and the positive cells were counted.The adipogenic differentiation abilities of MSCs from the different tissues were assessed.Results The MSCs from different tissues were spindle-shaped adherent cells.The FCM results showed the surface marker of MSCs were positive for CD44,CD73,CD90, and CD105,and negative for CD14,CD34,and CD45.The FT-MSCs displayed higher proliferative  ability than UC-MSCs and PL-MSCs（P<0.01）.The  proliferative index (PI) of FT-MSCs (36.66%±1.30%) was significantly higher than  those of UC-MSCs (18.23%±1.10%) and PL-MSCs (10.03%±1.20%)(P<0.01).The lipid droplets were found after induction,and the results of Oil red O staining were positive in all kinds of MSCs. However the adipogenic capacity of UC-MSCs was significantly higher  than those of PL-MSCs and FT-MSCs(P<0.01).Conclusion UC-MSCs,PL-MSCs and FT-MSCs have similar biological characteristics and cell surface markers.But FT-MSCs have the strongest proliferative ability and activity,UC-MSCs have the strongest adipogenic capacity in vitro.

Abstract:Objective To observe the effect of Bupleurum Liver-Coursing Powder on the neuronal apoptosis and autophage in hippocampus of depression model rats,and to explore the mechanism of its anti-depression effect.Methods 60 male SD rats were randomly divided into control group,model group and drug intervention group (n=20).The depression model was produced by giving the rats chronic unpredicted mild stress.The rats in model group and control group received the same volume of normal saline,and the rats in drug intervention group received Bupleurum Liver-Coursing Powder solution on the basis of model.The depressional behavior was examined using sucrose preference test,tail-suspension test and Morris water maze. Flow cytometry was employed for the detection and quantification of the apoptotic cells in the hippcampus.The expressions of LC-3 and Beclin 1 were detected using Western blotting method.Results  The pencertages of sucrose preference of the rats in model group and drug intervention group were significantly lower,while the tail-suspension immobility time and the excape latency time were higher than those in control group (P<0.05);the apoptotic rates of cells,the ratios of LC3-Ⅱ/ LC3-Ⅰ and the expression levels of Beclin-1 were higher than those in control group (P<0.05).Compared with model group,the percentage of sucrose preference of the rats in drug intervention group was increased (P<0.05),the tail-suspension immobility time and the excape latency time were decreased (P<0.05),and the apoptotic rate of cells,the ratio of LC3-Ⅱ/ LC3-Ⅰand the expression level were also decreased (P<0.05).Conclusion Bupleurum Liver-Coursing Powder has significant anti-depression effect,which may be related to inhibiting the apoptosis and degrading the autophage of neuros.

Abstract:Objective To explore the effect of QizhiJiangtang Capsule on the insulin resistance(IR) in the diabetic rats, and to clarify the action mechanism.Methods The diabetes rat models were induced by high fat diet combined with STZ injection.The successful models of the rats were randomly divided into diabetes group (DM),ShenqiJiangtang Granule group (SQ) and  high (QJH),middle-(QJM),low (QJL) doses of QizhiJiangtang Capsule groups;at the same time control group (NC) was established.The drug concentrations in  high,middle and low-doses of QizhiJiangtang Capsules  groups were 1.35,0.68，and 0.34 g?kg-1 respectively;and the  concentration of ShenqiJiangtang Granule was 0.27 g?kg-1. After the diabetic model was established successfully,the rats were treated for 8 weeks on the basis of drug dose.Then the levels of fasting blood glucose (FBG),fasting insulin (FINS),insulin resistance index (IRI) and biochemical indexes related to lipid metabolism of the rats were measured using blood glucose detector and automatic biochemistry analyser.The gene expression of insulin receptor substrate-1 (IRS-1),phosphatidyl inositol 3-kinase (PI3K),and glucose transporter 4 (GLUT4) in liver tissue were examined by Real Time PCR.The levels of tumor necrosis factor α (TNF-α) and adiponectin (ADPN) in serum were detected using ELISA.Results Compared with control group,the levels of FBG,FINS and IRI of the rats in diabetes  group were significantly increased (P<0.05 or P<0.01);the serum total cholesterol (TC),triglyceride (TG) and low density lipoprotein (LDL) levels were significantly increased (P<0.05),while the serum high-density lipoprotein (HDL) level was significantly decreased (P<0.05);the mRNA expression levels of  IRS-1,PI3K and GLUT4 in liver tissue were decreased (P<0.05);the level of serum TNF-α was increased (P<0.05),but the ADPN level was decreased (P<0.05). Compared with diabetes group,the FBG level and IRI of the rats in QizhiJiangtang Capsule and ShenqiJiangtang Granule groups were significantly decreased (P<0.01);the levels of FINS of the rats middle and high doses of in QizhiJiangtang Capsule  groups and ShenqiJiangtang Granule group were significantly decreased (P<0.05);the levels of serum TC,TG and LDL of the rats in  middle dose of QizhiJiangtang Capsule group and ShenqiJiangtang Granule group were significantly decreased (P<0.05 or P<0.01),but the HDL level was increased (P<0.05);the mRBA expression lvels  of  IRS-1,PI3K and GLUT4 inliver tissue were increased (P<0.05);the levels of serum TNF-α of the rats in   middle dose of QizhiJiangtang Capsule group and Shenqijiangtang Granule group were significantly decreased (P<0.05),but the serum ADPN levels were increased (P<0.05).Conclusion QizhiJiangtang Capsule can significantly improve the IR in the diabetic rats,and the pharmacological mechanisms are related to adapting the blood lipid component and insulin signal transduction pathways.

Abstract:Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method.Use the Sevage method to remove the protein in the crude polysaccharide.The structural characteristics of the polysaccharide were determined by FT-IR spectra.The  RM-1 and HeLa cells were divided into  control group and  different concentrations (0,50,100,200,300,400 and 500 mg/L) of ginseng polysaccharide groups.The survival rates of the cells in various groups were detected by MTT method.The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg/L) on the cancer cells were determined by CTL test.Results The extraction rate of ginseng polysaccharide  was 8.76% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between   different concentrations of ginseng polysaccharide groups after treated for 24 h compared with  control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly(P<0.05) compared with  control group; with the increase of ginseng polysaccharide concentration,the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg/L,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Abstract:Objective To explore the effect of trimetazidine on myocardial structure injury induced by pyran adriamycin and to clarify the protective effect   of trimetazidine on.the changes of  myocardial structure and its mechanism.Methods 36 Wistar rats were randomly divided into control group,model group and treatment group.The rats in model group and treatment group were injected with pyran doxorubicin 2.5 mg/kg(concentration 2 g/L)　by the caudal vein  once a week.The rats in control group were injected with  equivalent normal saline  for 6 weeks.The  rats in treatment group were intragastricly infused with  trimetazidine 5.4 mg/kg/d one day before making the model.The rats in control group and model group were injected with equivalent normal saline  for 8 weeks.At the end of the experiment,the  myocardial enzymes in serum of the rats in various groups were measured.The morphology of myocardium tissue was detected by light microscope and electron microscope.Results  Compared with model group,the levels of myoglobin,troponin I and alanine transaminase(ALT) of the rats in treatment group were significantly decreased（P<0.05）.Under  light microscope the  myocardium of the rats in model group   arranged disorderly,the structure was severely damaged,emyocardial was seen，the myofilament was dissolved;the myocardium of the rats in treatment group  arranged in order，the structure was nearly integrated，partial dissolution and fracture were found.Under  electron microscope in model group the myocardial muscle bundle dissolved,fractured and  disappeared,and the mitochondria was decreased, and the cytoplasmic matrix cavitation was seen;the  cardiomyocytes sarcomeres of the rats in treatment group arranged in order, local myofilaments were reduced slightly,the  surrounding mitochondria were oval and arranged in parallel between the muscle bundles.Conclusion Trimetazidine has protective effect on the  cardiomyocyte injury caused by pyran adriamycin,and its mechanism may be related to decreasing the injury of mitochondria and myocytes.

Abstract:Objective To explore the influence of icotinib in the apoptosis  of the human salivary adenoid cystic carcinoma cells ACC-M,and to clarify the mechanism of icotinib for the treatment of salivary adenoid cystic carcinoma.Methods The ACC-M cells were randomly divided into  controlgroup,2,4,8 μmo1/L^-1 icotinib  groups,p38-MAPK inhibitor SB203580(20 μmol/L^-1)   group, SB203580(20 μmol/L^-1)+4 μmo1/L^-1 icotinib  group;the cells were collected 4 h after treatment.The viability of ACC-M cells was measured by MTT assay．The apoptosis of ACC-M cells was assessed by caspase-3 activity kit．The expression of p-p38-MAPK protein was determined by Western blotting analysis.Results  Compared with control group,the  inhibitory rates of growth  of the ACC-M cells  in icotinib  groups  were significantly decreased(P<0.05),and the activities of caspase-3 were increased（P<0.05),and the expression levels  of p-p38-MAPK   were significantly increased（P<0.05）.Compared with 4 μmo1?L-1 icotinib  group,the expression level of p-p38-MAPK in SB203580+icotinib  group were decreased（P<0.05）,and the activity  of  caspase-3 was decreased dramatically （P<0.05）.Conclusion Icotinib may induce the apoptosis of ACC-M cells through the activation of p38-MAPK signaling pathway.

Abstract:Objective  To investigate the association between the minisatellite polymorphism in the first exon of PLA2G4C gene and schizophrenia,and to reveal the
important role of DNA sequence polymorphism in the pathogenesis of schizophrenia.Methods The minisatellite polymorphisms in the first exon of PLA2G4C gene in 91 patients with schizophrenia (case group) and 81 healthy persons(control group) were detected with PCR-sequencing analysis.The chi-square (χ2) goodness-of-fit test
 was used to analyze the distribution of  the PLA2G4C minisatellite polymorphism in various groups and to explore the association between the minisatellite polymorphism in
 the first exon of PLA2G4C gene and schizophrenia.Results There were minisatellite polymorphisms in PLA2G4C gene.Three kinds of polymorphisms 1×27 bp，2×27 bp
and 3×27 bp were found by sequencing.The distribution of  allelic frequencies at PLA2G4C polymorphism showed no statistical significance between case group and control group (P>0.05).No statistically significant difference was found in 3-homozygous haplotypes in PLA2G4C gene between case group and control
 group (P>0.05).At the same time,there was no statistically significant difference between 3-heterozygous haplotypes in PLA2G4C gene between case
 group and control group(P>0.05).Conclusion The minisatellite polymorphisms in the first exon of PLA2G4C gene are found,but the minisatellite polymorphism in the first exon of PLA2G4C gene may be not associated with the occurrence of schizophrenia.

Abstract:Objective To study the expression of tumor stem cell marker aldehyde dehydrogenase  1(ALDH 1) in invasive bladder cancer tissue and to clarify its relationship with the biological behavior of bladder cancer.Methods The ALDH1 expression in 109 cases of primary invasive carcinomas specimens (case group) and 20 cases of  normal bladder tissue surrounding cancer (control group) was detected by immunohistochemistry.At the same time,the ALDH1 expression in 6 cases of metastatic pelvic lymph node  tissue and  20 cases of non-metastatic pelvic lymph node tissue was detected.The relationship between the ALDH1 expression and the chinicopathological charateristics of invasive bladder cancer and its influence in the survival rate and disease-free survival were analyzed.Results The positive rates of ALDH1 expression in bladder cancer tissue and normal bladder tissue were 33.94%（37/109）and 5.00%（1/20），respectively, there was significant different between them（P<0.01）;they were 19.05%(8/42)and 43.28%(29/67) in the cases with non muscle invasive and nmuscle invasive bladder cancer, respectively,there was significant difference(P<0.01）;they were 13.04%(3/23) and 39.53%(34/86) in the cases of bladder cancer with low grade and high grade,  respectively,there was significant difference（P<0.05）;they were 50.00%(3/6) and 12.90%(4/31) in the tissue of bladder cancer with metastatic lymph nodes and non metastatic ones, respectively,there was significant difference（P<0.05）;they were 50.00%(3/6)and 0.00%(0/20)in the metastatic lymph nodes and non metastatic ones, respectively,there was significant difference（P<0.01）.The overall survival rate in the patients  with positive ALDH1 expression was 64.9% while it was 84.7% in negative ones,there was significant difference（P<0.05）;the disease-free Survival was 51.4% and 75% in the patients with positive and negative ALDH1 groups, respectively, there was significant difference（P<0.05）.
Conclusion The high expression of tumor stem cell marker ALDH1 is associated with staging，grading and prognosis of invasive bladder cancer.ALDH1 may play a role in the tumorigenesis,progression and metastasis of bladder cancer.

Abstract:Objective To identify the normal breast tissue and breast fibroadenoma  tissue by shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS),
and to explore the biological characteristics of FD and the identification method by discussing its spectroscope characteristics.
Methods The frozen section of 26 patients (all female,aged 19-59 years) were obtained by routine surgical resection.9 cases of normal tissue and 17 cases of breast fibroadenoma tissue were detected by Raman spectroscopy and then SHINERS technique was utilized.A total of 243 Raman and 273 SHINERS spectra were ob
tained.All the spectra were dealt with baseline corrected by fitting and subtracting a third-order polynomial and then smoothed with a 15-point Adjacent-Averaging.
Results The characteristic peaks of normal breast tissue appeared in 1 090,1 157,1 262,1 300,1 442,1 658,1 745,and 1 874 cm^-1.After adding SHINs,
some peaks shifted in 2-3 cm-1,the relative strengths of 1 090 and 1 157 cm^-1  were significantly increased,and the 1 496 cm-1 characteristic peak appeared.The main characteristic peaks of breast fibroadenoma  appeared in 751,880,930,880,1 262,1 442,1 579,1 658,and 1 745 cm^-1;one of the dominant characteristic peak should belong to lipids,but it can be seen that amide Ⅰ characteristic peak of protein became more significant.Conclusion Raman spectra can discover the differences of the characteristic peaks of amide Ⅰ between breast fibroadenoma  and normal breast tissues.By virtue of different enhancement effects of SHINs to Raman specific peaks of the various tissues,breast fibroadenoma can be distinguished from normal tissue successfully.
 

Abstract:Objective To observe the expressions of α-catulin and E-cadherin in tongue squamous cell carcinoma (TSCC)  tissue,and to explore
 their relationship with the occurrence and development of TSCC.Methods The expressions of α-catulin and E-cadherin in 55 cases of TSCC tissue and 10 cases
 of normal tongue tissue were examined by immunohistochemistry SP method.The relationship between the expressions and the clinicopathological characteristics
 and the relevance of the expressions of α-catulin and E-cadherin in TSCC tissue were analyzed.Results The positive expression rates of α-catulin in TSCC tissue and normal tongue tissue were 69.09% and 20.00%, respectively,and there was   difference between them (P<0.01).The expression of α-catulin was correlated to the histological differentiation,clinical stage and lymph node metastases of TSCC (P<0.05).The positive expression rates of E-cadherin in TSCC tissue and normal tongue tissue were 38.18% and 80.00%, respectively,and there  was significant  difference between them (P<0.01).The expression of E-cadherin was correlated to the clinical stage and lymph node metastases of TSCC ( P<0.05).There was a negative correlation between the expressions of α-catulin and E-cadherin in TSCC tissue（r=－0.466，P<0.01).
Conclusion The expressions of α-catulin and E-cadherin may be associated with the occurrence and development of TSCC,and they could be used as the parameters which predict the malignant degree and prognosis of TSCC.

Abstract:Objective  To investigate the expressions of metastasis suppressor 1 (MTSS1) and calcium activated protein 43 (Cap43) in esophageal squamous cell carcinoma（ESCC） tissue,and to clarify the relationship between the expressions of MTSS1,Cap43 and  the clinicopathological features of ESCC.
Methods 80 cases of ESCC tissue and 30 cases of normal adjacent-cancer tissue were collected,and the protein and mRNA expressions of MTSS1 and Cap43 in ESCC
tissue and normal tissue were detected by streptavidin-perosidase (SP) immunohistochemistry and RT-PCR;their relationships with the clinicopathological features  of ESCC were analyzed. Results The positive expression rate of MTSS1 in normal esophagus  tissue was significantly higher than that in
ESCC tissue detected by SP (83.3% vs 21.3%,P<0.01) and  RT-PCR (0.703±0.085 vs 0.295±0.065,P<0.01),However,the positive expression rate of Cap43
 in normal esophagus  tissue was significantly lower than that in ESCC tissue by SP method (16.7% vs 76.3%,P<0.01) and by RT-PCR (0.236±0.052
vs 0.693±0.078,P<0.01).The mRNA expression levels of MTSS1 and Cap43 in ESCC tissue were significantly related with the invasive extent,histological differentiation,TNM stage,and lymphatic metastases（P<0.05）of ESCC,but not related with the age,sex,tumor size and pathological type （P>0.05）.The mRNA expression of
MTSS1 was negatively correlated with the expression of Cap43 (r=－0.457,P<0.05).Conclusion The low-expression of MTSS1 and over-expression of Cap43 in ESCC tissue may contribute to tumor invasion and metastasis;the imbalance of MTSS1 and Cap43 may be one of the mechanisms of tumor invasion and metastases.

Abstract:Objective To investigate the differences in the structures,numbers and phenotypes of the dendritic cells (DCs) at different stages in colorectal cancer
(CRC) tissue, and to explore the effect of DCs in the occurrence and development of CRC.Methods 20 cases of CRC tissue were chosen as CRC
 group,other 20 cases of corresponding para-cancer tissue were chosen as control group.Immunohistochemical staining was used to detect the distributio
n,the structures,the number of positive cells at different stages;Western blotting was used to detect the expressions of CD1a and CD83.
Results The results of immunohistochemical staining showed that DCs scattered among glands or vessels in both groups;the CD1a+imature DCs (imDCs) were round and oval,dendritic processes were faintly visible in both groups,and the number of CD1a+imDCs in experimental group was significantly higher than that in control group (P<0.01).The CD83+ mature DCs (mDCs) were irregular,branched in both groups,and the number of CD83+mDCs in CRC  group was significantly lower than that in control group (P<0.01).The results of Western blotting method showed that the expression of CD1a protein was higher and the expression of CD83 protein was lower in CRC  group than  control group(P<0.01).Conclusion CD1a+imDCs may play important roles in promoting the occurrence and development of CRC;CD83+mDCs may inhibit the
 occurrence and development of CRC.

Abstract:Objective To evaluate the predictive values of ABCD,ABCD2,SPI-Ⅱ and ESSEN score
 models for the patients with   high-risk transient ischemic attack(TIA)  to develop to  cerebral infarction in short and long term.
Methods The   ABCD,ABCD2,SPI-Ⅱ and ESSEN scores of  235 cases of TIA patients were retrospectively analyzed.The incidence
of cerebral infarction was followed up for 7 d and 1 year,and the receiver operating characteristic curve (ROC) was drawn to calculate the area under  curve(AUC) to assess the accuracy of the score models,and compared with the original model and  the relative risk(RR) value was calculated.Results The 7 d-incidence and 1 year-incidence of cerebral infarction in the 235 TIA patients were 9.36 % and 20.43%．The AUC of ABCD,ABCD2, SPI-Ⅱ and ESSEN models for 7 d were 0.70,
0.74,0.67, and 0.62.The  AUC of 1 year were 0.62,0.62,0.64, and 0.65.Compared with the orginal models,the RRs for 7 d of ABCD score model of the TIA patients in low,middle,and high risk groups were 0.09，0.92，and 0.72；the RRs of ABCD2 score model were 0.49，0.59，and 0.65;the RRs of SPI-Ⅱ score model were 0.58，0.
87, and 0.55；the RRs of ESSEN score model were 0.11，0.18，and 0.55.Conclusion ABCD,ABCD2,SPI-Ⅱ and ESSEN score models
 can be  used to assess the risk of cerebral infarction after TIA in Chinese population.The  ABCD2 score model is of great value for short-term risk prediction,and the ESSEN score model is more value for long-term risk prediction.

Abstract:Objective To investigate the influence of early oral feeding (EOF) after laparoscopic surgery in the function status and gastrointestinal living qualit
y of the patients with colorectal cancer,and to clarify the feasibility of EOF after laparoscopic surgery.Methods Sixty-three patients underwent laparoscopic s
urgery of colorectal cancers participated in the trial.Of these,31 patients received EOF as EOF group,received a clear liquid diet on the first postoperative
 day followed by a regular diet as tolerated;the other 32 patients received traditional oral feeding (TOF) as TOF group who were fed with feeding only after the
 recovery of their postoperative gastrointestinal functions.The nasogastric tube was removed from all patients in both groups immediately after surgery.
Self-designed EOF questionnaire data,Karnofsky Scores and Gastrointestinal Quality of Life Index(GIQLI) Scores were used to evaluate the functional status and gastrointestinal living guality of the patients.Results The using time of  total parenteral nutrition (TPN)，time of postoperative hospital stay，and costs after surgery in EOF group were lower than those in TOF group(P<0.05);but there were no significnat differences in the  first passage of flatus and feces time between two groups(P<0.05)，also there were no significant differences in the incidence of nasogastric tube reinsertion,pulmonary infection,intestinal obstruction,balance of intestinal bacteria,fistula，incision infection between two groups (P>0.05)，and the incidence of abdominal distension was higher than that in TOF group（P<0.05）；on postoperative day 7，the albumin recovered faster in  EOF group（P<0.05），and on postoperative day 4 and 7，the pro-albumin also recovered faster in  EOF group（P<0.05）；the patients in EOF group had a higher Karnofsky score（P<0.05） and GIQLI score compared with the patients in TOF group（P<0.05）.Conclusion EOF after laparoscopic surgery in the patients with  colorectal cancers is beneficial for rehabilitation,and it can reduce the risk of hospitalization and saving its costs；it plays an active role in protein recovery，and  improves the functional status and gastrointestinal living quality of the patients.

Abstract:Objective To compare the X-ray cephalometric  results of teeth and skeleton relationship between individual lingual orthodontic and labial straight w
ire orthodontics,and to provide reliable theoretical foundation for selecting orthodontic system in clinic.Methods The general conditions of the patients,classification of malocclusion,severity of denture crowding and orthodontic design were used as the selection criteria.Thirty familiar adult patients with malocclusion in clinic were divided into two groups randomly: individual lingual orthodontics group (A group) and labial straight wire orthodontics group (B group),15 cases in each group.X-ray cephalometrics were taken using Steiner and Tweed cephalometrics analysis,the results of analysis were compared between  pre-treatment and post-treatment in two groups respectively,and the course of treatment in two groups was compared,too.The difference of treatment effect and course of treatment between two different orthodontic systems was evaluated.Results The relationship of first molar and canine of the patients in two groups was neutral relationship，overbite and overjet of anterior teeth was shallow,the teeth arrangement was orderly and no space was left after treatment.There was no statistically significant difference(P>0.05) between two groups in each measured values detected by Steiner Analysis after orthodontic treatment；there was no statistically significant difference(P>0.05) between two groups in each measured value detected by Tweed Analysis after orthodontic treatment；and there was no statistical significance(P>0.05) in the D-values of Steiner Analysis and Tweed Analysis between two groups before and after treatment.The duration of treatment of the patients in two groups had no  significant difference (P>0.05). Conclusion Both individual lingual orthodontics and labial straight wire orthodontics could obtain well teeth arrangement and occluding relationship and achieve satisfying clinical orthodontic outcome.

Abstract:Objective To analyze the sentinel lymph node metastasis in the breast cancer patients after neoadjuvant chemotherapy (NAC) and axillary lymph node di
ssection (ALND),and to investigate the application value of sentinel lymph node biopsy (SLNB) in the breast cancer patients after NAC.
Methods 52 breast cancer patients after NAC and 35 non-NAC breast cancer patients were selected.All of the patients accept SLNB and ALND and received a subareolar injection of 99m Technetium(Tc)  labeled unfiltered sulfur colloid.The sentinel lymph node was detected by a gamma probe during the operations.Then the conventional ALND were performed.All of the sentinel lymph nodes and axillary lymph nodes were sent for pathological  examination.Results A total of 113 sentinel lymph nodes were detected a fter NAC in 52  breast cancer patients,the successful identification rate was 100%.The accuracy,false negative rate,sensitivity,specificity,positive predictive value and negative predictive value of SLNB in the breast cancer patients after NAC were 94.2%,8.1%,91.9%,100%,100%,and 83.3%,respectively. A total of 73 sentinel lymph nodes were detected in 35  non-NAC patients,the successful identification rate was 100%. The accuracy,false negative rate,sensitivity,specificity,positive predictive value and negative predictive value of SLNB in the breast cancer patients after NAC were 94.3%,7.1%,92.8%,100%,100%,and 77.8%,respectively.Conclusion
 SLNB after NAC may gradually replace ALND as the standard treatment for lymph node negative patients after NAC treatment.

Objective To explore the effects of salmeterol/fluticasone propionate complicated or combined with N-acetylcysteine on the pulmonary function and  arterial blood gas analysis of the patients with chronic obstructive pulmonary disease(COPD)，and to evaluate its curative effect.Methods 84 cases of COPD patients were randomly divided into combination treatment group (n=44) and simple treatment group (n=40).The patients in combination treatment group were treated with
salmeterol/fluticasone propionate combinated with N-acetylcysteine while the patients in simple treatment group were treated only with salmeterol/fluticasone prop
ionate，both of which were followed up for 6 months.The changes of pulmonary function (FEV1%FVC,FEV1%Pred,PEF daily variation rate: Δ PEF%) and  theart
erial blood gas analysis indexes ( PaO₂ and PaCO₂ ) of the patients in two groups were recorded before treatment,3 months after treatment,and 6 months after treatment.Results The FEV1%FVC,FEV1%Pred and PaO2 of the patients in combinationtreatment group and simple treatment group were obviously increased 3 and 6 months after treatment compared with before treatment (P<0.01)，and the PaCO₂ were obviously decreased (P<0.01).And there were significant differences of the indexes mentioned above of the patients  between two groups(P<0.05)；but the indexes of each group had no significant differences between 3 months and 6 months after treatment(P>0.05).The Δ PEF% of the patients in two groups had no significant differences between inter-group and intra-group before and after treatment (P>0.05).Conclusion Combination of salmeterol/fluticasone propionate and N-acetylcysteine can obviously improve the pulmonary function and arterial blood gas analysis indexes  of the COPD patients,which is superior to the simple application of salmeterol/fluticasone propionate and has definite and lasting curative effect on the treatment of COPD.
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Objective To evaluate the value of the new ovarian function marker serum anti-Mullerian hormone(AMH) and to clarify the effect of  hysterectomy  on the ovarian function of the younger women.Methods The serum AMH,follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels  in 35 women suffered uterus benign lesion aged 36-45 years and  35 women suffered hysteromyoma without operation were measured at different time.And the ovarian arterial blood flow resistance index (RI) was measured by Doppler ultrasound.Results Compred with before operation,the serum AMH levels of  the patients in hysterectomy group 2 d and 3 months  after operation were  significantly decreased (P<0.05 or P<0.01).Compared with control group,the serum AMH levels of the patients in hysterectomy group 2 d and 3 months after operation were also decreased(P<0.05).The  ovarian arterial blood flow RI of the patients  in hysterectomy group 1 and 3 months after operation were increased compared with control group(P<0.05).There were no significant differences in the  serum FSH and LH levels  in  each  group between different time points(P>0.05).Conclusion Hysterectomy can affect  the ovarian function,and the  serum AMH level 3 months after operation is decreased and the  ovarian arterial blood flow RI is increased.AMH is  superior to FSH or LH in evaluating the changes of ovarian function.

Objective To investigate the clinical effects of different doses of propranolol in treatment of infantile mixed and deep hemangioma,and to provid theoretical and experimental evidence for  the treatment of hemangioma.Methods The selected 62 patients with  mixed and deep hemangioma were divided randomly into  low dose(1.5mg?kg-1?d-1)　 and high dose(3.0　mg?kg-1?d-1) of propranolol groups,3 times a day,6 months as a course,the changes in hemangioma size and the incidence of adverse reactions  were recorded.Results The efficacy  was evaluated using Achauer system.The total effective rate was 80.65% in low dose of propranolol group and 93.55% in high dose of propranolol group,including  6 cases of class Ⅰ(poor),9 cases of class Ⅱ(moderate),11 cases of classⅢ(good),and 5 cases of class Ⅳ(excellent)in low dose propranolol group；while  2 cases of class Ⅰ(poor),4 cases of class Ⅱ(moderate),10 cases of   class Ⅲ(good) and 15 cases of Ⅳ(excellent)in high dose of propranolol group.The efficacy in  high dose of proprandol group  was significantly better than that in  low dose of proprandol group(P<0.05)，there were no significant differences in the efficacies between different sites and different types(P>0.05) and there was no significant difference in the  incidence of  adverse reaction between  two groups (P>0.05).Conclusion The efficacy of oral 3.0 mg?kg-1?d-1 propranolol in treatment of infantile mixed and deep hemangioma  is  increased significantly,and there is no significant adverse reactions after increasing doses.Therefore,high dose of propranolol should  be recommended in order to improve the therapeutic effect.

Objective  To explore the changes of  cognition and sleep status in the youngsters with  internet addiction disorder (IAD),and to clarify  the relation between  cognition and sleep-related indicators of IAD  youngsters. Methods Event related potential(ERP)and Pittsburgh Sleep Quality Index(PSQI) test were used for forty-two IAD(case group) and forty non-IAD (control group)in youngsters,and the indexes of period latency(PL) of N1,P2,N2,P3 and amplitude of P3 (P3amp) of ERP were analyzed,the factor scores of sleep quality(SQ),sleep latency(SL),sleep time(ST),sleep efficiency(SE),sleep dysfunction(SD),sleeping pills(SP),daytime dysfunction(DD) and total score of PSQI were analyzed.The correlation  between the 　P3PL,P3amp of ERP and all the  indexes of PSQI in case group was analyzed.
Results The PL of N2,P3 of ERP in  case group was significanly longer and the  P3amp was significantly lower than those in  control group（P<0.01）;the factor scores of the SQ,SL,ST,SE,SD,DD, and the  total score of PSQI test had significant differences between  two groups（P<0.01）.The correlation analysis results showed that there were  remarkable positive correlations between the   P3 PL and the factor seores of the SQ,SL,ST,SE,SD,DD  and the total score of PSQI test in case group(r=0.46，0.34，0.51，0.40，0.48，and 0.54;P<0.01)，and there were  remarkable negative correlations betweeb  P3amp and the  indexes of PSQI test  in case group(r=－0.42,－0.45，－0.49，－0.38，－0.38，and －0.50;P<0.01).ConclusionThe IAD  youngsters have cognition and sleep dysfunction,and the both factors can affect each other.

Objective To compare the  long-term efficacies of anterior cervical decomprd fusion (ACDF) and conservative therapy in treatment of  unisegmental cervical disc herniation and to analyze the degree of   the long-term adjacent segment disc degeneration in  ACDF.Methods 120 patients treated in our hospital from January 2006 to January 2009 were selected,60 patients underwent ACDF as operation group and 60 patients underwent physical therapy and drug(conservative therapy) as non-operation group.All the  patients were recorded when they were diagnosed with cervical unisegmental cervical disc herniation,and followed-up for 60 months,and they were valuated with  American Spinal Injury Association(ASIA) score,Visual Analogue Scale (VAS),Japanese Orthopedics Association(JOA) score and imaging(Miyazaki grading system).Results107 patients were followed up,55 patients treated by operation were followed up for an average 59.7 months (59.7±0.4),while 52 patients treated by conservative therapy were followed up for an average 58.8 months (58.8±1.5).Compared with before treatment, theASIA, JOA,and VAS scores of the patients in two group after treatment were improved　(P<0.05),and the scores of ASIA,VAS,and JOA in operation group were superior to that in non-operation group (P<0.05),and  adjacent segment disc degeneration occurred more frequently in the upper adjacent segment than the lower adjacent segment (P<0.05）.Conclusion ACDF is superior to conservative therapy in alleviating symptoms and improving prognosis of unisegmental cervical discherniation.Long-term postoperative disc degeneration mainly occurrs in the upper segment.
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Objective To observe the incidence of acute radiation pneumonitis(RP) after  intensity modulated radiation therapy,and to explore the  relationship between the incidence of RP and the parameters from  dose-volume histogram(DVH)in the patients with non-small cell lungcancer(NSCLC).Methods By Pinnacle 7.6c TPS，46 patients with NSCLC   were treated by IMRT.The values of  the percentages of the lung volumes receieved 5,10,20,and 30 Gy radition to the total lung volumes (V5 ,V10,V20,V30) and mean  lung dose(MLD)  were  observed，and its correlation with acute RP was analyzed.Results The incidence of acute RP of the patients with NSCLC was 37.0%(17／46），among then there were 29 cases (63.0%) of Grade 0 (non-pneumonitis group),12 cases  (26.1%) of Grade 1,5 cases(10.9%) of Grade 2, and there were no Grade 4 and  5 RP. The values of V5 ,V10 ,V20 ,V30,and MLD  of lung were 53.34 %,43.12%,24.15%,15.36 %,and 16.02 Gy.The values of V5 ,V10 ,V20 ,V30,and MLD   in  pneumonitis group were 57.81％,48.91％,31.34％,17.83％,21.71 Gy，and they were 49.81％,39.78％, 21.82％, 13.12％,and 13.71 Gy  in non-pneumonitis group,there were significant differences between two groups(P<0.05).Conclusion The intensity modulated radiation therapy in treatment of NSCLC can protect thelung tissue,and the parameters of DVH such as  V5 ,V10 ,V20 ,V30 and MLD can predict the occurrence of RP.

Objective To explore the ultrasound imaging characteristics of thyroid papillary carcinoma complicated cervical lymph node tuberculosis,and to elucidate
the key points of ultrasound diagnosis and to distinguish with cervical lymph node metastasis of papillary thyroid carcinoma.Methods In total,15 well-documented cases of papillarythyroid carcinoma diagnosed definitely were selected,and there were 6 cases of concomitant lymph node metastasis.The ultrasonography of lymph node enlargement was analyzed,and the differences of the ultrasonographic characteristics between lymph node tuberculosis and metastatic lymph node including the location,swelling,calcification,blood flow and regional nodal liquefaction.Results Thyroid papillary carcinoma complicated with cervical lymph node tuberculosis was often  found in the areas of Ⅲ,Ⅳ and Ⅴ,especially in the area of Ⅴ.Variety of echo was mixed in tuberculous of lymph node,and the echo was inhomogenous.The tuberculosis of lymph node calcification was patchy inhomogeneous distribution.The echo in part of liquefaction of lymph node tuberculosis was cottony weak.The flow signal of tuberculous lymph appeared the surrounding or internal punctate distribution,and the soft tissue was echogenic and disorder around the lymph node tuberculosis.Conclusion When ultrasonography examination is performed in the patients with the thyroid papillary carcinoma complicated with cervicallymph node enlargement, the history should be considered to analyze the ultrasound characteristics to dignose by observing the lesions of the surrounding soft tissues.
 

ObjectiveTo obtain the  Hiwi gene encoding full-length and construct human Hiwi adenoviral vectors carryinggreen luorescence protein(GFP),and to establish  foundation for a further study on Hiwi function and mechanism of inducing leukemia stem cell differentiation and apoptosis.Methods All coding areas of human Hiwi gene full length were amplified using method of overlapping extension PCR technology，and  the full length coding aeras were inserted into the vector of Flag-IRES
-hrGFP carrying   GFP with Gateway technology to construct pDown- Hiwi-3×flag-IRES-hrGFP.The cloning vector pDown-Hiwi-3×flag-IRES-hrGFP and expression vector pAV.Des1d were used for homologous recombination reaction to obtain recombinant adenovirus vector pAV.Ex1d-Hiwi-3× flag-IRES-hrGFP.The positive clones were selected by PCR to extract the recombinant adenovirus plasmid and to pack into recombinant Ad-Hiwi-3×flag-IRES-hrGFP adenovirus.Results The human recombinant Hiwi was successfully cloned and the recombinant adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was found to be successfully constructed via restriction enzyme digestion and sequencing methods.The adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was transfected into the HEK293A cells using lipofectamine mediated gene transfection method.Under fluorescence microscope,the transfected cells with green fluorescence could be observed.Conclusion The expression plasmid of adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP is successfully constructed and  it can express in HEK293A cells.