Abstract:Objective
To explore the effect of schisandrin B(Sch B) on expression of vascular endothelial growth factor (VEGF) in glioma SHG-44 cells and its inhibitory effect on cell growth and to explore the possible mechanism.Methods Glioma SHG-44 cells were separated to control and 3 treatment groups,and then treated with 0,50,100 and 200 mg•L^-1  Sch B for 24-72 h.MTT assay was used to evaluate the proliferation of glioma SHG-44 cells.The levels of VEGF protein expression after treated with Sch B were measured by both ELISA and immuncytochemistry.Results Compared with control group ,the proliferation activity of glioma SHG-44 cells was slightly increased after  treated with 200 mg•L^-1 Sch B for 24 h(P<0.05);the proliferation abilities of the SHG-44 cells  were obviously decreased after treated with  50,100 and 200 mg•L^-1  Sch B for 48 h or 72 h;the VEGF protein level in the supernatant of cultured cells after treated with Sch B for 72 h was significantly decreased (P<0.01).After the glioma  SHG-44 cells were treated with Sch B for 72 h,the positive expression rates of VEGF protein in 50,100 and 200 mg•L^-1 groups were markedly reduced compared with control group (P<0.01).Conclusion Sch B can inhibit the expression of VEGF protein and growth of glioma SHG-44 cells,it suggests that Sch B may exert anti-glioma effect by inhibiting angiogenesis in glioma.

To observe the expressions and the co-localization of estrogen receptor alpha(ERα) and metastasis-associated 1(MTA1) in gastric cancer cells,and to explore the pathogenesis of gastric cancer.Methods Total protein was extracted from BGC823 and SGC7901 gastric cancer cells.The expressions of endogenous ERα and MTA1 were proved by Western blotting.The co-localization of endogenous ER α and MTA1 in the gastric cancer cells  was observed by using laser scanning confocal microscope.Results Two kinds of gastric cancer cells  BGC823 and SGC7901 expressed ERα and MTA1 detected by Western blotting,and they were co-localized in the nucleus of BGC823 and SGC7901 observed under laser scanning confocal microscope.Conclusion ERα  can  express in the gastric cancer cells  and co-localizated mainly in the nucleus with MTA.

To study the effects of broccoli polypeptide on the proliferation of C6 glioma cells of rats,and to discuss the induction of broccoli polypeptide on apoptosis of tumor cells.Methods The crude broccoli peptide was ectracted,separated and purificated.The rat C6 glioma cells were cultivated and  treated with different doses of broccoli polypeptide (0,0.01,0.10,1.00,10.00 mg•L^-1) for 24,48 and 72 h,and the apoptosis of cells was observed under inverted microscope,then the growth of C6 glioma cells after treated with broccoli polypeptide  was detected by MTT assay.Results  Broccoli polypeptide had  four main eluting peaks,and the second one (partⅡ) was rich in polypeptide which was used for continued study.The apoptotic bodies were found  under inverted microscope after the C6 glioma cells were treated with broccolipolypeptide,especially in 10.00 mg•L^-1 group.The MTT results showed that after treated  for 48 h,the proliferation ability of cells  in 10 mg•L^-1 group (0.127±0.011) was significantly decreased compared with control group(0.501±0.035,P<0.01).After treated for 72 h,all doses of polypeptide inhibited the growth of cells;compared with control group(0.753±0.022),the proliferation abilities of the cells in 1.00 mg•L^-1 group (0.583±0.082) and 10.00 mg•L^-1  group (0.073±0.001) were significantly decreased(P<0.01).Conclusion Broccoli polypeptide may induce the apoptosis of C6 glioma cells,and the effect is increased with the prolengation of the treatment time and the increasing of drug dose,which suggesting
 that broccoli polypeptide can  induce the apoptosis of tumor cells.

To observe the therapeutic effect of transplantation of mesenchymal stem cells(MSCs) on acute liver failure(ALF) in mice,and to provide a theoretical basis for further clinical application of MSCs.Methods The MSCs of mice were isolated,cultivated,pu
rified and identified in vitro.The model of ALF induced by  CCl4 was established.The ALF  mice were randomly divided into control and treatment groups.The MSCs were transplanted into tail veins of mice in treament group,and saline was transfused into the tail veins of mice in control group.The levels of serum ALT,AST and TBIL were determined.The pathological changes of liver tissues were observed by microscope after stained with HE.The survival rate was defined as the percentage of surviving animals at any specific time points.Results The cultured MSCs at G0-G1 stage accounted for 75.27% detected by flow cytometry.The expression of CD44 in MSCs was positive(88.71%),while the expression of CD34 was negative.After the MSCs were transplanted to ALF model rats,the survival rate in treatment group(65%) was higher than that in control group(25%).The levels of serum ALT,AST and TBIL were decreased after transplantation of MSCs compared with control group(P<0.01).The degrees of  liver cell necrosis and inflammatory cell infiltration in treatment group were lighter than those  in control group on day 1 and 2.And the pathological improvement  of liver in treatment group was more evident than that in control group on day 7.The expression level of ALB in treatment group was higher than that in control group on day 7  detected by immunohistochemistry.Conclusion Transplantation of MSCs can improve liver function and increase survival rates obviously of mice with ALF,which indicates that transplantation of MSCs has an obviously therapeutic effect on ALF in mice

To study the apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL) in non-small cell lung carcinoma H596 cells,and to explore the role and mechanism of apoptosis.Methods Non-small cell lung carcinoma H596 cells were cultivated,and acid phosphate assay was used to detect the apoptotic rate,and the expressions of caspase-8,Bcl-2 and Fas-associated death domain (FADD) proteins were detected by Western blotting in H596 cells.Results  The apoptotic rates of H596 cells were decreased after the cells were treated with 0.01-0.03 μg•L^-1 TRAIL,and from 0.10 μg•L^-1 TRAIL,the apoptotic rates were increased;the differences in apoptotic rates of H596 cells after treated  with 10.00-100.00 μg•L^-1 TRAIL were significant compared with control group(P<0.05).The Western blotting results showed that the expressions of caspase-8,Bcl-2,FADD proteins were normal.Conclusion High dose of TRAIL can induce apoptosis in H596 cells,and it may be  related to the normal expressions of   TRAIL related proteins.

To use embryo pancreas extract (EPE) from rats to induce pancreatic differentiation of human adipose tissue-derived mesenchymal stem cells(hASCs) into insulin-secretion cells, and to find new cell source for islet transplantation in clinic.Methods  The pancreatic micro-circumstance was built for hASCs to differentiate into　insulin-secretoin cells by extracting protein complex from new born SD rats.The hASCs in experiment group were induced by EPE and the cells in control group were cultured without EPE.20 d after induction,the expression levels of genes related with pancreas development  were detected by real-time PCR.ELISA  was used to detect the insulin secretion levels in experiment and control groups.The C-peptide expression was verified by fluorescent immunocytochemistry.Results The expressions of key markers of hASCs Oct 3/4 and Nanog were decreased in the process of EPE induction.On the 6th day after induction,the expression levels of POX-1 and CK-19 were up-regulated;and on the 12nd day,the insulin expression was positive.The insulin levels secreted by differentiated cells with and without glucose stimulation(401.09 mIU•L^-1±28.94 mIU•L^-1  vs   287.17 mIU•L-1± 15.67 mIU•L^-1,P<0.01) were much higher than that of undifferentiated cells (4.85 mIU•L-1±1.04 mIU•L-1,P<0.01).The C-peptide　positive cells　were found in protein by fluorescent immunocytochemistry in experiment  group.Conclusion hASCs have the potential to differentiate into pancreas cell lineages with EPE induction.They could become substitution of islet cells in treatment of diabetes mellitus.
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 To study the protective effect of gegecha (GGC) on liver injury in mice and to provide experimental basis for study on pharmacological effects of GGC.Methods 30 ICR mice were randomly divided into  blank group,vitamin(VC) group and  GGC group(100 mL•kg^-1),and all of the mice were treated once a day for 3 d.1 h after the last administration,all mice were intragastrically given with acetaminophen 1 000　mg•kg-¹.24 h later, the  number of dead mice was recorded in each group and the mortality was calculated.Another 60 ICR mice were randomly divided into control group,model group,VC group,and low,middle,and high doses of GGC groups,and there were 10 mice in each group.The mice in control group and model group were intragastrically given with distilled water,the mice in VC group were intragastrically given with VC (1 000 mg•kg^-1),the mice in GGC groups were intragastrically given with different doses of GGC(50,100,200 mL•kg^-1).All of the mice were treated once a day for 7 d.5 h after the last administration,the mice were intragastrically given with 500 mg•kg^-1  acetaminophen,except control group;24 h later,all of the mice were killed and the serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured according to kit introduction.The liver weights of donor were weighed,and the pathological changes of liver tissues were examined by HE staining method.The activity of superoxide dismutase (SOD) was measured by  xanthin oxidase method and the content of malondialdehyde (MDA) was measured by  thiobarbituric acid　method.Results The mortality in  blank group was 80%,40% in VC group,and 50% in  GGC(100 mL•kg^-1) group.The protective rates of VC and GGC in dead mice induced by acetaminophen were 50% and 40%,respectively.The liver quotient  of mice in  model group was increased compared with control　 group(P<0.01),and the liver quotients of mice in  middle and high doses of GGC group were lower that in  control group(P<0.05). Compared with model group,the serum AST and ALT levels in mice  with iver injury were  significantly reduced in GGC groups,and showed a significant dose-dependent relationship.Compared with model group,the serum levels of AST,ALT and MDA were decreased (P<0.05),the SOD activities were increased in GGC groups (P<0.05 or P<0.01).The HE staining results showed that  liver cells were swelling and there was a large area of inflammatory infiltration in model group,while in GGC groups,the liver inflammatory infiltration was reduced with the increasing of GGC dose,and the effect of 200 mg•kg-1 GGC  was most obvious.Conclusion GGC has significantly protective effect on liver tissues in mouse liver injury models induced by acetaminophen,and the protective effect of GGC on liver tissues is more obvious with the increasing of GGC dose.
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To explore the effects of fluorouracil (Fu) combined with wortmannin in wtp53/mtp53 colon cancer cells (HCT-116 and SW-480) and to clarify　the role of p53 in proliferation and apoptosis of human colon cancer cells.Methods The HCT-116 and SW-480 cells in the logarithmic growth　phase were divided into Control,Fu,Fu+wortmanin and Fu+wortmanin+PFT-α groups separately,and the cells were treated  with Fu,wortmannin and PFT-α, respectively.The survival and apoptotic rates of HCT-116 and SW-480 cells at different time (12,24， and 48 h) in various groups were detected by MTT assay and flow cytometry (FCM) with Annexin V-FITC individually.Results Compared with Fu group,the survival rates of HCT-116 and SW-480 cells in Fu+wortmannin group were significantly decreased with a time-dependent decrease (P<0.05);FCM analysis showed that the apoptotic rates of HCT-116 and SW-480 cells were increased with the prolongation of time (P<0.05);compared with  Fu+wortmannin group,the  survival rate　of SW-480 cells  in Fu+wortmannin+PFT-α group at 48 h was significantly decreased(P<0.05),and the apoptotic rate was markedly increased(P<0.05),while the apoptotic and survival rates of HCT-116  cells had nosignificant change(P>0.05).Conclusion Wortmannin could enhance the inhibitory effect of Fu on the human colon cells (HCT-116 and SW-480);inhibition of p53 expression can enhance the inhibitory effect  of Fu combined with  wortmannin  on proliferation of mtp53 SW-480 cells and promote the apoptosis of mtp53 SW-480 cells.

To investigate the expression of calcium-sensing receptor (CaSR)  in pulmonary arterial smooth muscle  and hydrogen sulfide(H₂S)   in plasma of rats with high pulmonary blood flow  pulmonary hypertension (PH),and to clarify the role of both of them in the mechanism of PH.Methods 18 male SD rats were randomly divided into   control group and PH group.The left lung resection surgery was performed in rats in PH group.35 d after feeding,the right heart catheterization was used to measure mean pulmonary artery pressure(PAMP) of rats in  control group and PH group;the right ventricular weight / body weight (RV / BW) and right ventricle /(left ventricle + interventricular septum) ［RV/（LV+S)］ ratios were detected,and the changes of pulmonary vascular structure  were observed under  optical microscope.Spectrophotometry was used to detect the plasma H₂S content.Real-time quantitative PCR was performed to detect the CaSR mRNA expression in pulmonary artery endothelial cells of rats in tow groups.Results The PAMP,the RV/BW and RV/(LV+ S) ratios in PH group were significantly higher than those in control group (P<0.05).Under light microscope,the pulmonary vascular muscle level was increased and the lung relative medial thickness of middle and small muscular arteries was also increased significantly in PH group(P<0.05).The CaSR mRNA expression in pulmonary artery endothelial cells in PH group was significantly higher than that in control group (P<0.05);the plasma H₂S content in PH group was significantly decreased compared with control group(P<0.05).Conclusion The changes of the CaSR mRNA expression in pulmonary artery endothelial cells in high pulmonary blood flow condition may be involved in PH  by changing the generation of endogenous H₂S.
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Abstract:Objective  To explore the effects of diammonium glycyrrhizinate on bleomycin induced pulmonary fibrosis (PF) and to further clarify the underlying fibrotic mechanisms.Methods 36 Wistar rats were randomly divided into normal control group,BLM-induced pulmonary fibrosis group(model group) andglycyrrhizinate group.The pulmonary fibrosis models of rats were established by injecting   BLM via trachea (5 mg/kg once).From the 2nd day after injection,the rats in glycyrrhizinate group were intraperitoneally injected with glycyrrhizinate(75  mg/kg/d),while the rats in other two groups with normal saline injection instead.On the 28th day,all rats were sacrificed,the blood samples were obstained to measure inducible nitric oxide synthase (iNOS) and nitric oxide (NO) levels.Their left lung lobe tissues were removed and used for pathological section examination.Results  The pulmonary tissue of the rats in  normal control  group was normal，and the pulmonary fibrosis of the rats in glycyrrhizinate group was significantiy lighter,and the pulmonary fibrosis of the rats in model group was severe.Compared with normal control group，the serum iNOS and NO levels of rats in model group were significantly increased（P<0.01）,but the serum iNOS and NO levels of rats in glycyrrhizinate group had no changes（P>0.05）；the serum iNOS and NO levels of the rats in glycyrrhizinate group were lower than　those in model group（P<0.05 or P<0.01）.Conclusion  Diammonium glycyrrhizinate can significantly reduce the serum levels ofiNOS and NO in BLM-induced pulmonary fibrosis rats and  alleviate pulmonary fibrosis.  

Objective To study the effects of resistant starch(RS) on intestinal fermentation in diet-induced obese (DIO) rats，and to clarify the possible mechanism of RS to improve intestinal function.Methods  60 SD rats were divided  randomly into 2 groups.One of the groups was selected as control group,and the rats were fed with normal foods.The rats in the other  group were fed with high-fat foods for 7 weeks.The DIO rats were divided into five groups,and were fed,for 5 weeks,respectively with high-fat foods,hight-fat foods containing 5% RS,10% RS,15% RS,and 10% RS plus dextran sulfate(DS) as inhibitor to butyric acid.Then,small intestinal and cecal tissue and cecal contents were collected,and body weight gain,the ratio of fat to body,cecal area and weight,amount of short chain fatty acids（SCFA）and pH in cecal contents were determined.Results Compared with control group,the body weights,ratio  of fat to body and pH of cecal contents of the rats in 10%RS group and 15%RS group were decreased(P<0.05 )；the cecal area  in 15%RS group was increased(P<0.05).The acetic acid,propionic acid and butyric acid levels in faeces in 5%,10% and 15%RS groups were increased(P<0.01).The weights of cecal contents in 10%RS group and 10%RS+DS groupwere decreased （P<0.05）.The acetic acid and propionic acid levels in faeces in 10%RS+DS group were increased （P<0.01）.The dose of RS was negatively related to the rat body weight (r=－0.862，P<0.01) and was positively related to the levels of acetic acid,propionic acid and butyric acid in cecal contents of rats(r₁=0.945，r₂=0.943，r₃=0.949，all P<0.01).Conclusion RS may significantly increase the cecal area，reduce the pH of cecal contents and increase short chain fatty acid levels in faeces.It is certified that RS can improve intestinal fermentation function of DIO rats.

Objective To analyze the survivin expressions in primary hepatocellular carcinoma (HCC) tissue and para-carcinoma tissue,and to investigate the eff
ects of survivin  on tumor blood vessel growth and inhibiting apoptosis of tumor endothelial cells.Methods The human HCC,para-carcinoma and normal liver tissues were collected,and the relationship between the positive rate of survivin protein expression and pathological parameters of HCC was  analyzed by immunohistochemitry,and the survivin protein expression levels in different tissues were detected by flow cytometry,and the survivin gene expression levels in different tissues were measured by quantitative real-time PCR( qRT-PCR).Results The survivin protein expression in HCC tissue was related to portal vein tumor thrombus formation (P<0.05),and had no relationship with tumor size,casple,AFP level and pathological stages of HCC (P>0.05).The flow cytometry results showed that the survivin protein expression in HCC tissue was higher than those in para-carcinoma and normal liver tissues (P<0.05); the survivin protein expression in para-carcinoma tissue was higher than that in normal liver tissues,but there was no  significant difference (P>0.05).The qRT-PCR results showed that the survivin mRNA was only detected in HCC tissue,not detectable in para-carcinoma or normal liver tissues.Conclusion Survivin is highly expressed in HCC tissue,but the survivin gene is not expressed in para-carcinoma tissue,and the  survivin protein may transport through a paracrine manner from HCC.

Objective To investigate the effect of hypoxia inducible transcription factor 1α(HIF-1α) on proliferation of rectal cancer HT-29 cells and to clarify its mechanism.Methods The wild type HT-29 cells(Wt-HT-29) and Si-HT-29 cells transfected with HIF-1α small interfering RNA  were cultivated  under normoxia and hypoxia conditions.The expressions of HIF-1α and hexokinase-Ⅱ(HK-Ⅱ) under normoxia and hypoxia conditions were detected by using Western blotting method．The survival rates of cells in various groups were detected by using MTT method.Furthermore，theATP content was detected by using ATP Kit.Results Western blotting showed that the  expressions of HIF-1α and HK-Ⅱ in Wt-HT-29 cells were higher than those in  Si-HT-29 cells under hypoxia condition. The amount of ATP and the number of  wild-type cells generated under normoxic conditions were regarded as 100%.The percentages of the ratios of the amount of ATP and the number of Si-HT-29 cells under normoxia condition and the amount of ATP and the number of the two cell lines under hypoxia condition to the  amount of ATP and  the number of  Wt-HT-29 cells under normoxia condition were regarded as the respective amount of ATP and the surrival rate of cells.Under normoxia condition the ATP amount and cell survival rate of HT-29 cels were 93.6% and 92.0%， respectively；and there  was no significant difference compared with Wt-HT-29 cells (P> 0.05).But under hypoxia condition the survival rates  of Wt-HT-29 cells  and Si-NT-29  cells were 85.0% and 61.3%，respectively,and there was significant difference(P<0.05).At the same time the ATP amounts  of Wt-HT-29 cells and Si-HT-29 cells were 81.2% and 42.9%,respectively;and  significant difference was observed (P<0.05).Conclusion HIF-1α can improve ATP production of rectal cancer cells by inducing HK-Ⅱ expression to increase the proliferation,and it may become a new therapy target of rectal cancer.

Abstract:Objective To observe the changes of transport protein Farnesoid X receptor (FXR) and bile salt export pump (BSEP) in liver cells of obstructive jaundice rats,and to discuss the influence of FXR activation and endotoxin (ET) on the expression of BSEP.Methods  Thirty-six healthy SD rats were randomly divided into sham operation group (Sham group),common bile duct ligation (CBDL) group and bile reflow (BR) group.The FXR and BSEP transcription and protein levels of hepatocytes were measured with average optical density (AOD) and immunohistochemistry microscope in liver tissues from the three groups at each time point.Meanwhile,the levels of serum total bilirubin(TBIL),total bile acid (TBA) and ET were examined with conventional methods.The relationships between FXR,TBA,ET and BSEP were analyzed by linear correlation analysis.Results The serum levels of TBIL,TBA and ET of rats in sham group at each time point after operation had no change(P>0.05);the serum levels of TBIL,TBA and ET of rats in CBDL group after CBDL were significantly increased(P<0.01);the serum levels of TBIL,TBA and ET of rats in BR group after BR were gradually decreased,the levels of TBIL and TBA 7 d after BR had no difference compared with sham group(P>0.05),but the ET level was also higher than that in sham group(P<0.05).The expression levels of FXR in  sham,CBDL and BR groups were increased gradually and slowly,but no significant differences were found among three groups (P> 0.05).The expression levels of BSEP in  sham and BR groups tended to decrease with time;the expression level of BSEP in  CBDL group was decreased firstly and then was increased,there was  no significant differences compared with  sham group (P> 0.05).Positive correlation was found between the genetic expression levels of FXR and TBA (r=0.303,P< 0.05),while negative correlation was found between the expressions of FXR and BSEP (r=－0.359,P< 0.05).The  expression levels of BSEP and ET were also negatively correlated (r=－0.412,P< 0.01).Conclusion The FXR level doesn’[KG-*3]t  play a leading role in the  expression of BSEP in the obstructive jaundice rats.The down-regulation of BSEP expression is mainly due to the expression level of ET.

Abstract:Objective To explore the in vitro and in vivo antibacterial activities of Impatiens Balsamina water extraction and to provide the pharmacological reference for seeking the new antibacterial drug. Methods The in vitro experiment was divided into negative control group (group A),penicillin control group (group B),streptomycin control group (group C),different doses of Impatiens Balsaminawater extraction (0.01,0.10,1.00 and 10.00 g/mL) groups(group D-G).The Impatiens Balsaminawater extraction was prepared by impregnation.The in vitro inhibitory effects of the Impatiens Balsamina water extraction on Staphylococcus aureus,Escherichia coli,Candida albicans,Proteus sp.,Pseudomonas aeruginosa,Bauman Acinetobacter，and Klebsiella pneumoniae were assayed by groove and paper disc antibacterial experiment.The in vivo experiment was divided into negative control group (group A),different doses of Impatiens Balsamina water extraction(0.5，5.0 and 50.0 g/kg) groups (group B-D).The in vivo inhibitory effects of Impatiens Balsamina water extraction were detected with Staphyloccocus aureus and Escherichia coli as index,the mice were given  Impatiens Balsamina water extraction by gavage for 7 d,then the mice were intraperitoneally injected with Staphyloccocus aureus and Escherichia coli suspension.The in vivo antibacterial activity of Impatiens Balsamina water extraction were detected by observing the mortality rate in mice.Results The in vitro assay showed that when the concentration was over 1　g/mL,the Impatiens Balsamina water extraction exhibited inhibitory effects on all the 7 kinds of pathogenic bacteria,the individual diameters of the inhibition zone were greater than that in negative control group and the difference was statistically significant (P<0.05).The in vivo assay showed that when the concentration was over 5　g?kg-1,the water extraction had antibacterial activity against Staphyloccocus aureus and Escherichia coli.The mortality rate of mice in Impatiens Balsamina water extraction group was significantly decreased (55.6%) compared with negative control group (88.9%).Conclusion Impatiens Balsamina water extraction contains the antibacterial ingredient.

Objective
To detect the expressions of hypoxia inducible factor -2α and ATP-binding cassette super family G2(ABC G2) in  mammospheres(MSs)  transplantation tumor tissue  in  nude mice，and to discuss the  possible  mechanism of the MSs’ strong tumor formation ability.Methods The  MSs cultured in serum-free media supplemented with growth factors were transplanted in the nude mice,and the growth of thetransplantation tumor was recorded.The expressions of HIF-2α and ABCG2 in transplantation tumor tissue were observed by immunohistochemistry.The expressions of HIF-2α and ABCG2 potein and mRNA in transplantation tumor tissue were detected by Western blotting and Real-time RT-PCR.Results The  MSs had the stronger tumor formation ability than the MCF-7 cell lines. Immunohistochemistry showed the expressions of HIF-2α and ABCG2 in transplantation tumor tissue in  nude mice. The expressions of HIF-2α and ABCG2 proteins in MSs transplantation tumor tissue were higher than those  in  MCF-7 transplantation tumor tissue and  para-tumor tissue（P<0.05）.The expressions of HIF-2α and ABCG2  mRNA in MSs transplantation tumor tissue were higher than those  in  MCF-7 transplantation tumor tissue  and  para-tumor tissue（P<0.05）.Conclusion  There are high expressions of HIF-2α and ABCG in  MSs transplantation tumor tissue.It is speculated that HIF-2α and ABCG may be the possible mechanism of MSs’ strong tumor formation ability.

Objective  To investigate the influence of phosphatase and tensin homologue deleted on chromosome ten(PTEN) in invasion of tongue squamous cell carcinoma  and to clarify the relationship between PTEN and epithelial-mesenchymal transition (EMT) in tongue squamous cell carcinoma. Methods  The SCC-4（non-transfection group）,pEGFP-PTEN-SCC-4（transfection group） and pEGFP-SCC-4（empty vector group）were cultivated at the same time. The cell invasion was observed by Transwell assay.The expressions of E-cad，snail and vimentin proteins  were examined by Western blotting.Results The number of the cells which passed the membrane were:82±5,80±4 and 42±5 in non-transfection group，empty vector group and transfection group 36 h after culture in Transwell assay;there was no significant difference between non-transfection group and empty vector group (P>0.05),but there was significant difference between non-transfection group and transfection group (P<0.001).Compared with non-transfection group,the E-cad expression was up-regulated(P<0.05),and the expressions of vimentin and snail were down-regulated(P<0.05) in transfection group;but there was no significant different between non-transfection group and empty vector group (P>0.05).Conclusion PTEN may  reduce the invasion of tongne squamous cell carinoma SCC-4 cells by inhibiting the EMT process of tumor cells.

Objective  To establish a  method to separate and identify the  rat renal tubular epithelial cells (RTECs) and analyze the proliferation  ability of the obtained cells,and to provide theoretical basis and technique support for analysis experiment in vitro of kidney diseases such as  kidney injury and so on.Methods The renal tubules were separated with   type Ⅱ collagenase digestion  and purified with Percoll density gradient centrifugation and cultured in normal environment,when the RTECs grew out from the tubule,the cytokeratin-18 (CK-18) expression was detected with immunohistochemistry and the expressions of CK-18 in P0,P1 and P2 cells were detected with flow cytometry.The proliferation ability  of each generation was evaluated with CCK-8 method.Results  After cultured for 24 h,the RTECs began to grow outside from the tubule and they looked like paving road stones after cultured for 72 h.In the process of passage,the expression efficiencies of CK-18 were 95.9%,91.5%,and 76.8% in P0,P1 and P2 RTECs,and  the RTECs were dead gradually. The proliferation ability  of  P2 cells was more weaken than that of P1 cells (P<0.05),and more mixed cells existed in the culture system.Conclusion The method established in this study is cheap,credited as well as convenient.However,the cells just could be passaged for 2 times and couldn't be cultured for long time.Only the cells of P0 and P1 could be applied in analysis experiment in vitro.

Abstract:Objective  To observe the influence of red clover flavone from golden buckwheat(RCFGB) in migration ability in vitro of human gastric cancer SGC7901 cells,and to explore its mechanism. Methods  Ethanol extraction was used to prepare RCFGB.The gastric carcinoma SGC7901 cells administrated with different doses (5 and 10　g?L-1) of RCFGB and PBS solution were used as experimental groups and control group.Scarification test and  Transwe Ⅱ test were used to observe the influence of different doses of RCFGB in migration ability of  human gastric cancer SGC7901 cells under the same culture conditions;Western blotting method was performed to detect the effects of different doses of RCFGB on the interleukin 6 ( IL-6 ) protein expression levels in SGC7901 cells of rats.Results Compared with control group,the healing abilities of scarification of SGC7901 cells in 5 and 10 g?L-1 RCFGB groups were apparently weakened (P<0.05),and the number of transmembrane cells were decreased significantly (P< 0.05),and the expression levels of IL-6 proteins in SGC7901 cells were significautly decreased(P<0.05).Conclusion  RCFGB can inhibit the migration ability of human gastric cancer SGC7901 cells,and its role may be related to  inhibiting the IL-6 protein expression level in SGC7901 cells.

Abstract:Objective  To explore the association between single nucleotide polymorphisms (SNPs) of forkhead box transcription factor O1 (FOXO1) and the susceptibility to type 2 diabetes mellitus (T2DM) in the Han population of Northern China,and to clarify  the association between  SNPs of FOXO1 gene and T2DM.Methods Using case-control study design,190 cases of T2DM patients were recruited in  case group and 193 healthy persons were selected as control group.The method of Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALI-TOF-MS) was applied to detect the SNPs  of rs2755213,rs2701880,and rs10507486 in FOXO1 gene.Results There were no significant differences in the frequencies of the alleles and the genotypes at rs2701880,rs2755213 and rs10507486 sites between two groups(P>0.05).There were no significant differences in the distribution of  the haplotypes rs2755213-rs2701880,rs2701880-rs10507486 and rs2755213-rs2701880-rs10507486 formatted by rs2701880,rs2755213 and rs10507486 sites between two groups(P>0.05).Conclusion The polymorphisms of  rs2701880,rs2755213 and rs10507486 in FOXO1 gene may not be the biomarkers of susceptibility to T2DM in Han population of Northern China.

Abstract:Objective  To explore the inhibitory effectiveness of  tacrolimus(Tac)  and ciclosporin(Csa) on acute rejection  in patients after kidney transplantation  and to provide guidance for the selection of immunosuppression medicine after kidney transplantation.Methods Computer searches were in Medline and  CNKI．The randomized controlled trials (RCT) comparing Tac with CsA in treatment of  acute rejection after  transplantation were included．Data was extracted independently by two reviewers．The methodological quality was assessed by the Jadad Score．Statistical analysis was performed with RavMan 5.0．Results 226 studies were found at first，and 31 RCTs comparing Tac with CsA in treatment of  acute rejection after kidney transplantation were identified at last．Meta-analysis showed that  2 and 14 studies about  biopsy-proved acute rejection during half a year and 1 year after transplantation  were included.The morbidities  of acute rejection during half a year and 1 year after transplantation in Tac group were  lower than those in CsA group with statistical difference,half a year RR 0.49，95%CI(0.37，0.67)，P<0.01;1 year RR 0.65，95%CI(0.56，0.75)，P<0.01.For steriod-resistant acute rejection 4 and 7 studies were included during half a year and 1 year after transplantation. The morbidities of acute rejection during half a year and 1 year after transplantation in Tac group were lower than those in CsA group with statistical difference,half a year RR 0.41，95%CI(0.30，0.57)，P<0.01; 1 year RR 0.54，95%CI(0.37，0.78)，P=0.01.Conclusion The effectiveness of Tac in inhibiting acute rejection after kidney transplantation is better than CsA.
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Abstract:Objective   To study the differential expression of miRNAs and their target genes in papillary thyroid carcinoma（PTC）tissue,and to analyze the potential roles of miRNAs as biomarkers and in carcinogenesis of PTC. Methods The PTC samples and their matching normal thyroid tissues were examined and collected.The genes and miRNA expression profiles were examined with Illumina Bead Chips.KEGG pathway was used to analyze the function of differential expression genes.MiRNA target genes were predicted by implementing three computational analysis programs,the results were verified by qRT-PCR.Results 248 miRNAs and 3 631 genes were found to be differentially expressed(gene:P<0.05,miRNA:P<0.01) in PTC tissues as compared with their matching normal thyroid tissues.Hsa-miR-101（target gene:ITGA3）was identified.The hsa-miR-101 expression in PTC tissue(59.8%) was down-regulated and ITGA3s expression (100%)  was up-regulated compared with their matching normal thyroid tissues.Conclusion ITGA3 may be the  target gene of hsa-miR-101,and both of them may play important roles in occurrence and development of PTC.

Abstract:Objective  To approach the effectiveness and safety of zaleplon compared with zopiclone in treating  insomnia with Meta-analysis and to provide basis for their application in clinic.Methods Computer　search was performed using the Cochrane Central Register of Controlled Trial，Ovid-medline,PubMed,EMBASE,CNKI,WanFang,VIP，and the randomized controlled trials (RCTs) were collected,then the retrieved studies according to predefined inclusion and exclusion criteria were screened,the quality of included studies was evaluated and RevMan 5.1.2 software was used for Meta analysis. Results  A total of 315 articles were found and 15 of them were finally included.The evaluation of the effectiveness of  insomnia in treatment of insomnia  was performed with sleep disorders scale reduction rate,χ2=7.54，df=14，P=0.91，Z=1.21(P＝0.23）；95%CI 0.88［0.71，1.08］. Security side effects of drug safety scale was used to evaluate the security,χ2=9.71，df=14，P=0.78，Z=4.09(P<0.0001）；95%CI　0.66［0.54,0.81］.Conclusion Zaleplon has the same effectiveness as zopiclone in treating insomnia and less side effect in safety compared with zopiclone.

Abstract:Objective  To clarify the relationship between MRI quantitative parameters and astrocytoma related  indexes by analyzing the correlation between MRI quantitative parameters and the expression level of hypoxia inducible factor-1α（HIF-1α） in cerebral astrocytoma.Methods  A total of 33 cases of astrocytoma proved by operation and pathology were collected.The MRI quantitative parameters,such as edema index(EI),relative signal intensity on Gd-enhanced T1WI(RSIGd) and enhancement percentage(EP) were measured.The expression of HIF-1α in tumor tissues was detected with immunohistochemical method.Results The values of RSIGd and EP of low grade astrocytoma(WHO grade Ⅰ-Ⅱ) were significantly lower than those of  high grade astrocytoma (WHO grade Ⅲ-Ⅳ) (P<0.01).The EI value of low grade astrocytoma was lower than that of high grade astrocytoma(P<0.01).There was no expression of HIF-1α in normal brain tissue.But HIF-1α expressed in astrocytoma with different grades.The expression level of  HIF-1α  in low grade astrocytoma was lower than that in high grade astrocytoma(P<0.001).The expression level of  HIF-1α  was positively correlated with the value of RSIGd( r=0.431,P<0.05).The expression level of  HIF-1α  was significantly positively correlated with the value of EP (r=0.651,P<0.001).There was no obviously significant correlation between the level of  HIF-1α  and the value of EI (r=0.317,P>0.05).Conclusion  RSIGd and EP are very helpful to evaluate the pathological grades and the permeability of new vessels of cerebral astrocytoma.High grade cerebral astrocytomas are often accompanied by peritumoral edema but it can’t be explained by only one theory.

Abstract:Objective  To compare the stress distribution of the residual root of first mandibular molar between the split post-crown rehabilitation system and split post-core-crown rehabilitation system with 3-dimensional finite element,and to offer  proper parameters for clinical application.Methods  An excised first mandibular molar was chosen to get a CT scanning in the area of its residual root.By reverse engineering software MIMICS and model reconstructing software MAGICS,the 3-dimensional model of such rehabilitation systems were built.The features of a model,such as material properties,contact reactions,restrains,and loads were set under the software of ABAQUS.Results  The stress analysis results showed that under vertical loads,the post-crown rehabilitation system appeared a stronger stress concentration at the contact position between the post and the root canal than the post-core-crown rehabilitation system(P<0.05);meanwhile,under lateral loads,a significant stronger stress concentration appeared in the post-crown rehabilitation system of the point between the crown and the post compared with the post-core-crown rehabilitation system (P<0.05).On the same position of a model,the force on root canal caused by lateral loads was greater than that of vertical loads.Under the same amount of loads,the root canal orifice suffered a greater force than internal part of the root canal,and there was  significant difference(P<0.05).The offset analysis results of the crown indicated that under vertical loads,the maximum displacements of the two systems were both 0.01 mm,and there was no significant difference(P>0.05).While under lateral loads,the post-core-crown had a greater maximum displacement than the post-crown,and there was  significant difference(P<0.05).Conclusion  When applying the split post-crown rehabilitation system,several measures should be take to enhance the resistance performance of the root canal orifice and reduce the force that residual roots suffer in order to get the prosthodontic treatment better.

Abstract:Objective To explore the effects  of CXCR4 / SDF-1 axis and the VEGF  on pathogenesis of ankylosing spondylitis (AS) and their related mechanisms,and to clarify the reliability of using VEGF as the index of active stage and monitoring prognosis of AS.Methods Flow cytometry (FCM) was used to detect the expression of CXCR4 on peripheral blood mononuclear cells (PBMCs) of 30 AS patients and 20 healthy persons.Enzyme linked immunosorbent assay (ELISA) was used to determine the serum VEGF and the SDF-1  levels in two groups.Relationship among CXCR4,SDF-1 and VEGF,and the relationship between above three indexes and disease activity indexes such as erythrocyte sedimentation rate(ESR) and  C reactive protein(CRP) were analyzed.Results The positive expression rate of CXCR4 of the patients in AS group was significantly higher than that in control group (P<0.05).The serum VEGF and SDF-1 levels of the patients in AS  group were significantly higher than those in control group (all P<0.05).There were positive correlations among the CXCR4,SDF-1 and VEGF (r=0.828,r=0.638,r=0.723,all P<0.05).There were positive correlations between the serum VEGF  level and the levels of ESR and CRP (r= 0.584，r=0.723,all P<0.05).But there was no obvious correlation between the CXCR4,SDF-1 serum level and the levels of ESR and CRP ( all P>0.05).Conclusion CXCR4/SDF-1 response axis and VEGF may have decisive roles in estimating the occurrence,development,severity and prognosis of AS;VEGF can be as reliable index of active stage and monitoring prognosis of AS,while those three may be the principal targets of targeting therapy.
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Objective  To observe the short-term efficacy of GP regimen in adjuvant chemotherapy in patients with non-small-cell lung cancer，and to study t
he factors influencing efficacy and analyze the adverse reactions.
Methods  Clinical data of 68 NSCLC patients received postoperative adjuvant chemotherapy of GP regimen was  retrospectively analyzed.Kaplan-Meier method was used to draw the disease-free survival curve.Log-rank time series analysis was used between different subgroups;C
OX proportional hazards model was used for the univariate and multivariate analyses.
Results  Of all the 68 cases，the median recurrence-free survival time was 33.7 months，56 cases were followed up more than one year，and one year disease-free survival rate was 83.9%;25 cases were followed up more than two years，and
two-year disease-free survival was 72.0%.Univariate analysis suggested that clinical stage(P<0.05)，pathological type(P<0.05),degree of differentiation(P<0.05) significantly influenced the short-term efficacy. Multivariate analysis suggested that clinical stage(P<0.05),degree of differentiation(P<0.05) were thei
ndependent factors of the short-term efficacy.The major adverse reactions were stage 3 to 4 myelo-suppression including neutropenia（33.9%），granulocytopenia（20.6%），anemia（4.4%），and thrombocytopenia（4.4%），and  stage 3 to 4 nausea and vomiting(10.3%).
Conclusion The GP regimen is feasible,well-tolerated with a high disease control rate in one or two years in the adjuvant chemotherapy;clinical stage
and degree of differentiation are the independent factors of the short-term efficacy.

Objective  To investigate the incidence of postoperative delirium after using dexmedetomidine and sub-anesthetic dose of ketamine and to clarify their influence in the incidence of postoperative delirium in elderly orthopedic patients under total intravenous anesthesia.
Methods  One hundred and twenty elderly patients aged more than 60 years underwent elective orthopedic surgery,ASA
Ⅰ-Ⅲ,were randomly divided into 4 groups(n=30): normal saline (control group),ketamine group,dexmedetomidine group,ketamine+dexmedetomidine group.The patients in ketamine group received an intravenous injection of ketamine  at a dosage of 0.5 mg?kg-1.In dexmedetomidine group the patients received dexmedetomidine at a dosage of 1  μg?kg-1 by intravenous injection  before induction of anesthesia followed by a continuous infusion at0.5  μg/kg/h till 30 min before the end of operation.In ketamine+dexmedetomidine group,the patients received both 0.5 mg?kg-1 ketamine  and  1.0  μg/kg dexmedetomidine over 10 min by  introvenous injection followed by a continuous infusion of dexmedetomidine at 0.5 μg/kg/h till 30 min before the end of operation. The patients in control group were administered with the same amount of normal saline.The blood sample
s were taken before anesthesia,at the end of operation and 24 h after operation.The level of serum interleukin-6 (IL-6) was detected using enzyme-linkedim
munosorbent assay (ELISA).The Confusion Assessment Method (CAM) was applied to evaluate postoperative delirium 1 h,1 d and 3 d after operation.
Results  There were no statistical significance in age,gender,weight,operation time,anesthesia time,anesthesia dosage,bleeding,urine volume and infusion of the patients between four groups(P>0.05).13 patients in the study developed delirium：8 patients in ketamine group (26.7%),2 patients in dexmedetomidine group (6.7%),3 patients in control group (10%);no delirium was found in ketamine+dexmedetomidine group. Compared with control group,the incidence rate of delirium in ketamine group was increased significantly (P<0.05).Compared with ketamine group,
the incidence rate of delirium in ketamine+dexmedetomidine group was decreased significantly (P<0.05).There was no significant difference in serumIL
-6 levels of patients between four groups(P>0.05).
Conclusion When they are applied together,dexmedetomidine could alleviate the side effects of ketamine and could decrease the incidence rate of delirium which mechanism is not responsible with inflammation.

Objective To observe the influence of the sub-anesthetic doses of ketamine and dexmedetomidine on early postoperative cognitive dysfunction(POCD) und
erwent orthopedic surgery in elderly patients,and to clarify its related mechanism.
Methods 120 patients aged 60 years underwent elective orthopedic surgery were randomly
divided into ketamine group,dexmedetomidine group,ketamine+dexmedetomidine
group and  control group,30 cases in each group.All patients received total intravenous anesthesia.Before anesthesia,
the patients in ketamine group received 0.5 mg?kg-1 ketamine intravenous injection.The patients in dexmedetomidine group received infusion of dexmedeto
midine 1 μg/kg at first,followed by  0.5 μg/kg/h infusion until 30 min before the end of operation.The patients in ketamine+dexmedetomidine group
received intravenous infusion of ketamine and dextromedetomidine.The patients in control group received intravenous infusion of saline.The Mini Mental State Examination (MMSE) was used to assess cognitive function 1 d before operation and 1,7 d after operation.The incidence of POCD was recorded.Blood samples were taken before anesthesia induction,at the end of operation and 24 h after  operation for
determination of IL-6.  Results Compared with control group,the time from consciousness
 to extubation of the patients in dextromedetomidine group was singnificantly prolonged(P<0.05).
 The incidence of POCD were 26.7% and 13.3% in control group at 1,7 d after operation,6.7% and 0 in ketamine group,
 20.0% and 10.0% in dexmedetomidine group, 13.3% and 3.3% in ketamine+dexmedetomidine group. Compared with  control grou
p,the incidence of POCD  in ketamine group at 1,7 d after operation was significantly decreased (P<0.05),the incidence of POCD in dexmedetomidine group and ketamine + dexmedetomidine group at 1,7 d after operation had no
   significant differences(P> 0.05).The serum IL-6 levels had no significant difference between various groups (P<0.05).
Conclusion The application of sub-anesthetic dose of ketamine can reduce the incidence of early POCD in orthopedic surgery.Dexmedetomidine alone or combined with ketamine can not reduce the incidence of early POCD in orthopedic surgery.The occurrence of POCD has nothing to do
 with inflammatory reaction.
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Abstract:Objective To determine the effect and safety of parecoxib sodium on pos
toperative analgesia in patients after renal transplantation,and to provide basi
s for application of analgesia medicine for patients after renal transplantation.
Methods Forty-eight patients underwent renal allogeneic transplantation were randomly
divided into parecoxib sodium group,fentanyl group,and control group,and there
were 16 patients in each group.The patients in three groups were given parecoxib
 sodium 40 mg,fentanyl 0.2 mg and saline 2 mL,respectively.The hemodynamic  parameters of patients in various groups were recorded at  5 min before e
xtubation(T1),extubation time(T2),5 min after extubation(T3) and 10 min after extubation(T4);and the occurrence of  cough,nausea and vomiting of patients
 were recorded;the postoperative pain and sedation levels were evaluated;the urine output,the levels of serum creatinine(SCr) and blood urea nitrogen (BUN) of
 patients in various groups before operation and 12,36 h after operation were compared.
Results There were no significant differences of operation time,intraoperative
infusion amount and hospitalization time of patients between various groups (P> 0.05).
Compared with T1 time point,the  SBP  levels of patients at T2,T3 and T4 in control group were increased(P<0.05);compared with T1 time point,the levels of oxygen sa
turation (SpO2)of patients at T3 and T4 in fentanyl group were decreased(P<0.05).There were no significant differences of
hemodynamic parameters of patients in parecoxib sodium group at T1,T2,T3 and T4 time points(P> 0.05).
Compared with control group,the incidence rates of tussis and dysphoria of patients in parecoxib sodium group and fentanyl
 group were decreased(P<0.05);there were no siginificant differences of the incidence rates of nausea and vomiting between var
ious groups(P>0.05).Compared with control group,the using times of tramadol hydrochloride  of patients in parecoxib sodium
group and fentanyl group were decreased.There was also no siginificant difference of urine volume,serum BUN,and SCr of
 patients between  various groups before and after operation (P>0.05).There was also  no siginificant difference of postoperative analgesia between fentanyl group and control group(P>0.05).The scores of postoperative analgesia of patients in parecoxib sodium group were lower than those in the other groups(P<0.05);the scores of postoperative sedation of patients in fentanyl group was lower than those in the other groups(P<0.05).
Conclusion Parecoxib sodium can be a candidate drug for postoperative  pain relief in patients underwent  renal transplantation,which has fewer side effects.

Objective   To explore the  relationship between glycemic control and implant healing
 in the patients with type 2 diabetes mellitus by studying the influence of diff
erent blood sugar levels in  the inflammatory indexes of implants and implant st
ability.Methods 21 patients with type 2 diabetes mellitus(diabetes group) and 10 healthy individuals (control
 group) were selected.40 implants were placed under local anesthesia, blood routine exam
ination was performed before and 24 h after operation,and the amount of inflammatory cells was counted.The glycated hemoglobin (HbAlc) levels
were assayed 8  and 12 weeks after implantation,the mean values were recorded as blood sugar indexes.The implant stability indexes were detected by means of
resonance frequency analysis on weeks 0,2,4,6,8 and 12 after implantation.The maximum decrease in
 stability from baseline and the time for stability level returned to baseline were calculated.
Results The inflammatory cell levels in diabetes group were higher than those in non-diabetic after implantation（P<0.05）,while no difference was found
before implantation（P>0.05）.The implant stabilities were compared according to HbAlc
levels;the patients with  higher HbA1c level had a greater maximum decrease in stability from baseline,while the difference between various groups had statistical significance（P<0.01）.The patients with HbA1c ≥7.6% required a longer time for stability level to returne
 to baseline than the patients with HbA1c ≤7.5%（P<0.01）.Conclusion The inflammatory reaction and implant stability in the patients with type 2 di
abetes mellitus are related to glycemic control.Better control of blood glucose will improve implant healing.

Objective  To observe the  clinical efficacy and safety of long-term sublingual immunotherapy with dermatophagoides farinae drops in ch
ildren with bronchial asthma  and to provide a safe and effective method to treat bronchial asth
ma in children.Methods  A two-year randomized and parallel-controlled study was conducted in
128 children (aged 4-14 years) with mild to moderate asthma symptoms.The patients
were randomly assigned to treatment group and control group(n=64).The patients in treatment group were treated with sublingual immunotherapy (SLIT) with dermatophagoides farinae drops.The patients in control group were treated according the manufacturer's instructions.The clinical efficacy was evaluated by monthly follow-up visits.After treatment for 6 months，1 year and  2 years using
 dermatophagoides farinae drops,the asthma symptom scores and PEF%  were evaluated.
Results Of the 128 children,5 cases dropped out before the study completion.After 6 months,1 year and  2 years of treatment,the allergic asthma symptom
 scores of the patients  in treatment group were significantly decreased compared with control group
 (P<0.05).After 6 months,1 year and 2 years of treatment,the PEF% of the patients in treatment group were
singnificantly increased compared with control group (P<0.05).There were no severe adverse events during the course of immunotherapy,except for mild reactions.
Conclusion Long-term  sublingual immunotherapy with dermatophagoides farinae drops can signi
ficantly improve symptoms and lung function in children with  asthma,and lower the incidence of adverse reactions,so it is effective and safe in the treatment
of children with bronchial asthma.

Objective To observe the clinical curative effects of compound diisopropylamine dichloroacetate (CDDI) in treating fatty liver disease induced by ta
moxifen(TAM) and to provide theoretical basis for its clinical application.Methods 72 cases of breast cancer patients with fatty liver disease induced by TAM were randomly divided into control group(n=38) and treatment group(n=34).The patients in control group were treated with   basic treatment alone.The patients in treatment group were injected intravenously  with CDDI 80 mg  once daily for 4 weeks in addition to basic treatment.The changes of clinical symptoms,liver function enzymes,blood lipids and liver imaging   of the patients in two groups were observed before and after treatment.The efficacy and safety of drugs were examined.Results After treatment，the total effective rates were 82.3% in treatment group and 23.7% in cont
rol group,there was significantly difference between two groups（P<0.01）.Compared with before treatment,the contents of alanine aminotransferase (ALT),total cholesterol (TC),and triglyceride (TG)of the patients in treatment group were significantly decreased after treatment（P<0.05）;and there also were  significant differences compared with control group after treatment(P<0.05).There were no significant differences of the
 contents of ALT,TC and TG in control group between before treatment and after treatment（P>0.05）.No severe adverse drug reactions were found in two groups during treatment period.Conclusion  CDDI could be used as a effective drug in the treatment of fatty liver disease induced by TAM.
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Objective  To quantitively assess the parameter alteration of renal blood flow in patients with diabetic nephropathy (DN)  with the contrast-enhanced ultrasound (CEUS),and to  evaluate the value of the technique  in diagnosis of early renal function changes of DN patients.Methods
 40 diabetic patients were equally divided into two groups according to Mogensen’[KG-*3]s staging criteria: normal albuminuria group(group Ⅰ) and
early DN group(group Ⅱ);and 15 cases of healthy volunteers were used as control group (N group)(n=15).All subjects were performed renal CEUS perfusion imaging,and QontraXt image analysis software was applied to select the region of interest (ROI) in the renal cortex.Then the time intensity curve (TIC) and kidney blood perfusion parameters were collected.Results  The renal blood perfusion was clearly shown in real time CEUS;compared with N group,the time to peak (TTP),regional blood volume (RBV),and mean transit time (MTT) of the patients in group Ⅰ were increased,there were significant differences (P<0.05);but there were no significant differences of derived peak intensity (DPI ) and regional blood flow (RBF ) between two groups (P> 0.05).Compared with group Ⅰ and N group,the RBV,TTP and MTT of the patients were increased,the DPI and RBF were reduced in group Ⅱ,there were significant differences 0.05).Conclusion The CEUS technical analysis can be used in evaluating renal abnormality of the DN patients in early period by showing the changes of renal perfusion parameters.
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Objective To understand the situation of smoking and quitting smoking and influencing factors by investigating smoking and quitting smoking of m
edical staff in Jilin Province,and to  provid key points and scientific information for the smoking control.Methods  By stratified sampling methods,3787 med
ical workers in 50 medical institutions of 9 districts(Changchun,Yanbian,Liaoyuan,Baishan,Baicheng,Zhenlai,Dunhua,Jingyu,Dongliao ) in Jilin Province were selected to undergo a questionnaire survey.The investigators came from medical institutions，and they investigated objects one by one after  training.The q
uestionnaires included smoking,smoking cessation and demographic information.SPSS 13.0 software was adopt to deal with the data.The data was described by rate and proportion,and  different rates were compared using  Chi-squaretest.The smoking and quitting smoking status  of medical staff in Jilin Provinceand influencing factors were analyzed.Results The total smoking rate of the medical staff in Jilin Province was 15.45%,the smoking rate of male was 44.20%,and the female’ was 0.40%.In the smoking people,the smoking rate in group ＞40 years old was the highest (22.06%),the smoking rate of the people with high school/technical school education was the highest (19.69%),and the smoking rate of the people in  Center for Disease Prevention and Control was the highest(24.82%).Among smoking-medical staff,44.71% ever tried to quit smoking and 39.04% wanted to quit smoking.To protec
t oneself and families from the smoking damage was the main influence factor of  smoke abatement.Conclusion  The smoking rate of  medical staff in Jilin P
rovince is still higher.And the  medical staff should accept the health education of smoking control to promote understanding of the smoking damage.

Objective  To prepare the FITC labeled Escherichia coli　(E.coli) O157∶H7 antibody with  high titer,high purity,high specificity,high fluorescence intensity
 and stability,and to approach its application in immunofluorescence detection.Methods Standard E.coli O157∶H7 strain was cultured and the bacteria was inactivated by adding formaldehyde solution to prepare vaccine.After four immunizations,blood samples were collected from the marginal ear vein.After purified by the method of caprylic acid-sulfide saturated solution precipitation and HiTrap Protein G chromatography column on AKTA Protein purification device,the purity and content of protein and titer of antibody were detected using the methods of SDS-PAGE,BCA Fit,indirect ELISA.Different ratios of FITC to protein and different reaction conditions were selected to label E.coli O157∶H7 polyclonal antibody,the F485/535 was measured and t-test was used to identify the best ratio of FITC to  protein and the best reaction condition.The reaction of polyclonal antibody with the mixed liquid of E.coli O157∶H7 and Staphylococcus aureus (S.aureus) was observed.Results The best E.coli O157∶H7 polyclonal antibody purity method of HiTrap Protein
 G was 35% and 100% gradient elution;the titer of antibody was 1∶25 600.It showed no cross-reactivities with 8 kinds of enterobacters such as Salmonella,Enterobacterc loacae and so on.The best ratio of FITC to protein was 1∶10,and the best reac
tion condition was 20℃and 2 h.When this fluorescence antibody was applied to react with E.coli O157∶H7 and S.aureus,it could react obviously and stron
gly.It had stability with E.coli O157∶H7 after irradiated with strong fluorescence  for 15 min,without across-reaction with S.aureus.
Conclusion FITC labeled E.coli O157∶H7 polyclonal antibody which high  purity,fluorescence intensity and stability is prepared  at t
he first time.The study  completes the major step of establishing of immunofluorescence detection method of E.coli O157∶H7.

Objective  To construct uncoupling proteins（UCPs）chimeras and mutations and provide conditions for study on the functional domains of uncoupling pro
tein family.Methods The gene fragments of UCPs chimera were obtained by routine polymerase chain reaction （PCR）and splicing overlap extension PCR（SOE-PCR）.Then these sequences were respectively cloned into the vector of pCR2.1 and sequenced.The recombinant eukaryotic expression plasmid pcDNA3.1 containing UCPs chimera was constructed by restrictive enzyme ligation.The recombinant plasmid of pcDNAUCP1UCP2,pcDNAUCP2UCP1 and pcDNAUCP3UCP1 were identified by PCR and restrictive enzyme digestion.The mutations of pcDNA-UCP3R167G/pcDNA- UCP3R
E171/172GG were generated by site-directed mutagenesis and then verified by sequencing.Results The identification of target gene by restrictve enzyme and PCR was the same as expectation.The sequence of chimera gene was completely identical with the expectation device.Conclusion
The recombinant eukaryotic expression plasmid containing UCPs chimeras and mutations is constructed successfully.