To  investigate the role of endoplasmic reticulum stress protein in oxidative stress-induced autophgy in human cervical carcinoma Hela cells,and to clarify the possible mechanism of endoplasmic reticulum stress-autophagy pathway in regulating the survival of tumor cells.Methods Hela cells were divided into control group,VK3 (30   μmol•L^-1) group，ER stress inhibitor TUDC (500  μmol•L-1) group，VK3(30   μmol•L^-1) +TUDC(500  μmol•L^-1)  group.The morphological changes of endoplasmic reticulum of Hela cells  were observed by transmission electron microscope.The survival rate of Hela cells in each group was measured by MTT assay.The expression levels of glucose-regulated protein-78 (GRP78) and LC3-Ⅱ were determined by Western blotting method.Results The morphology of Hela cells in control group was normal.The autophagic vacuoles and dilated endoplasmic reticulum were observed in cytoplasm of Hela cells in VK3 group.Compared with VK3 group，the survival rate of Hela cells in VK3+ TUDC group was decreased(P<0.05).Compared with control group,the expression levels of GRP78 and LC3-Ⅱ in VK3 group were decreased(P<0.05).Compared with VK3 group,the expression levels of GRP78 and LC3-Ⅱ in TUDC+ VK3 group were decreased (P<0.05).Conclusion Endoplasmic reticulum stress protein can mediate the  autophgy of Hela cells in oxidative stress-induced injury of Hela cells.
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To explore the effect of ［5-nitro-2-(3-phenylpropylamino)-benzoic acid］NPPB，a chloride channel blocker,on the growth of human glioma SHG-44 cells and to clarify  the potential mechanism of proliferation inhibition and apoptosis induction of NPPB on SHG-44 cells.Methods The glioma SHG-44 cells were cultured and divided into control and 50,100 and 200   μmol•L^-1 NPPB groups,and the influence of NPPB in proliferation of SHG-44 cells was　measured by MTT assay.The cell cycle and apoptotic rate were analyzed by Cell Analysis Machine.Results Compared with control group,the proliferation of SHG-44 cells was inhibited after treated with 50   μmol•L^-1 NPPB for 48 h (P<0.05),the proliferation of SHG-44 cells was improved after treated with 200   μmol•L^-1 NPPB for 3 h (P<0.05) and the proliferation of SHG-44 cells were inhibited significantly after treated with 100 and 200   μmol•L^-1 NPPB for 24 and 48 h (P<0.01).The detection  results of cell cycle showed that compared with control group,the percentage  of SHG-44 cells  in G1 phase was decreased and the percentage of SHG-44 cells in G2/M phase was increased(P<0.01) in 50   μmol•L^-1 NPPB group 24 h after treatment;Part of SHG-44  cells was arrested in G1 phase in 100 and 200   μmol•L^-1 NPPB groups 24 h after treatment（P<0.01）.The detection  results of apoptosis  showed that the apoptotic rates of SHG-44 cells were increased to 24.64% and 41.85% after treated with 100 and 200   μmol•L^-1 NPPB for 24 h, respectively,compared with control group (P<0.01).Conclusion High doses of NPPB can inhibit the proliferation and induce apoptosis of SHG-44 glioma cells,which indicates that chloride channels may play essential roles in the proliferation and apoptosis of human glioma SHG-44 cells.

To construct the recombinant eukaryotic expression vector of mitochondrial superoxide dismutase 2 (SOD2) and to explore its expression efficacy in human umbilical vein endothelial cells (HUVEC).Methods SOD2 gene full-length cDNA was amplified from HUVEC by RT-PCR and the target gene SOD2 was connected with the plasmid pcDNA3.0 vector.The recombinant plasmid was constructed and transfected into HUVEC.The cells were grouped into HUVEC,pcDNA3.0 and pcDNA3.0-SOD2 transfected groups.The expression of SOD2 in HUVEC was detected by RT- PCR.

To investigate the effects of antidepressant quetiapine on the apoptosis,necrosis and cell cycle of the hippocampus neurons injured by glutamate (Glu),and to demonstrate the mechanism of quetiapine on nervous system.Methods Primary cultured hippocampal neurons were injured with Glu.Normal control group,Glu injury group and quetiapine (0.1,1.0,10.0,50.0,100.0,200.0， and 500.0  μmol•L^-1 ) group were set up. The changes in apoptosis,necrosis and cell cycle of neurons were detected by FCM before and after administration with quetiapine.Results Compared with normal control group,the percentage of apoptotic neurons in Glu injury group was increasd (P<0.01);compared with Glu injury group，the apoptotic rates in 50.0-200.0  μmol•L^-1 quetiapine groups were decreased (P<0.05，P<0.01).The necrotic  rate in Glu injury group was increased to 2.9 times compared with normal control group (P<0.05 );compared with Glu injury group，the necrotic  rate of neurons in 0.1   μmol•L^-1 quetiapine group was decreased (P<0.05).Compored with normal control group,the percentage of neurons in G0/G1 phase in Glu injury group was increased (P<0.001),the percentage of of neurons in S phase was decreased (P<0.01),and the  percentage of neurons in G2+M phase didn’t change significantly. Compared with Glu injury group，the percentage of neurons in G0/G1 phase in 200.0   μmol•L-1 quetiapine group was decresed,which was  decreased to 68.9% of that in Glu injury group,and the percentage of neurons in S phase was increased to 3.4 times of that in Glu injury group (P<0.05).Conclusion Quetiapine can prevent the hippocampal neurons from the damage induced with Glu,decrease the apoptosis and necrosis,and change the cell cycle.

To investigate the effect of T cell factor 3(TCF3) gene silencing on the expression of Oct4 and to explore the mechanism of Oct4 expression regulation by TCF3 in mouse embryonic carcinoma F9 cells. Methods The TCF3 gene in F9 cells was silenced by RNAi technology and used as TCF3 silencing group;meanwhile blank control group (mock) and negative control group (scramble) were set up. The mRNA and protein expression levels of TCF3 and Oct4 in F9 cells in various groups were detected by RT-PCR and Western blotting methods.The expression of Oct4 in F9 cells was detected by immunofluorescence assay.Results Oct4 was highly expressed in nucleus of F9 cells.Compared with mock group and scramble group,the mRNA and protein expression levels of TCF3 in TCF3 silencing group were significantly decreased（P<0.01）,while the mRNA and protein expression levels of Oct4 were significantly increased（P<0.01）.There was no significant difference between mock group and scramble group（P>0.05）.Conclusion Gene silencing of TCF3 in F9 cells can obviously promote the transcription and expression of Oct4,which indicates that TCF3 can negatively regulate the expression of Oct4 in F9 cells as one of the inhibitors.

To investigate the interventional effect of puerarin (Pur) preconditioning  on focal cerebral ischemia-reperfusion(IR) injury in rats and to explore the possible therapeutic time window. Methods Fourty male Sprague-Dawley rats were randomly divided into  IR,Pur preconditioning(PC)-6h，PC-12h，PC-24h and PC-48h groups(n=8). 100 mg•kg-1 Pur was given at 6,12,24 or 48 h before cerebral ischemia in the rats in last four groups,0.9% saline 1 ml•kg-1 was injected ip at the end of 90 min ischemia in the rats in IR group. A focal cerebral ischemia reperfusion model was induced by the middle cerebral artery intraluminal suture method. All animals were subjected to  the left middle cerebral artery occlusion for 90 min，followed by reperfusion for 24 h．The neurological deficit score was measured at 24 h after IR. The brain infarct volume percentage(BIVP) was then assessed after 2％TTC staining following the last neurological outcome evaluation. Results  Compared with IR group,the neurologic deficit scores in PC-6h,PC-12h and PC-24h groups were statistically improved (P<0.01),but not in PC-48h group (P>0.05).The BIVP  in PC-6h,PC-12h and PC-24h group were significantly decreased compared with IR group (P<0.01).No statistically significant difference in BIVP was found between IR group and PC-48h group (P>0.05).There were no statistically significant differences in neurologic deficit score and BIVP between PC-6h,PC-12h and PC-24h groups(P>0.05).Conclusion Pur precondioning can protect rat brain against IR injury.The therapeutic time window of Pur neuroprotection may be over 24 h．

To construct the GST-tagged eukaryotic expression vectors of human p21-activated kinase 1(hPAK1) autoinhibitory domain(AID) with different activities and identify the expressions of their recombinant proteins,and to explore the function of PAK1 AID and molecular targeting role in tumor therapy.Methods The pcDNA3.1HisC-PAK1 full length plasmid was used as template,and the fragment of wild PAK1 AID was amplified with PCR，and the fragment of PAK1 L107F was amplified with long primer mutant method.The PCR fragments were double-digested and cloned into pEBG.The wild and inactive types of pEBG-PAK1 were transfected into HEK293 cells and their expressions were identified by Western blotting.Results The wild and in active PAK1 AID  fragments with 200 bp  were obtained by PCR.5 000 bp vector band and 200 bp PAK1 AID band were obtained by double-digestion.The wild and inactive types of  pEBG-PAK1  were expressed in HEK293 cells,and the molecular weight of the protein was 33 000.Conclusion The wild and inactive types of PAK1 AID  eukaryotic expression vectors are successfully constructed.The expressions of wild and inactive  types of PAK1 AID proteins  are identified.

To observe the influence of the “Warming yang and Nourishing qi” acupuncture treatment in the serum acetylcholine receptor antibody (AchR-Ab) and interferon-gamma (IFN-γ) expression levels in the rats with experimental autoimmune myasthenis gravis (EAMG) and to analyze the mechanism of acupuncture treatment to myasthenis gravis (MG).Methods AchRα1 129-145 peptide fragments were used to immune the female Lewis rats to establish EAMG  models.30 successful EAMG model rats were randomly selected and divided into acupuncture treatment group,drug control group and model control group (n=10);the unmodelled 10 rats purchased at the same period were used as blank control group.The rats in acupuncture treatment group were treated with “Warming yang and Nourishing qi” acupuncture treatment,30 min per time,once a day,7 d for a period,1 d rest between two periods,two periods of continuous treatment.The rats in drug control group were treated with  pyridostigmine lavaged treatment,18.5 mg•kg^-1•d^-1，continuous treatment of 15 d.The rats in the other two groups had no any special treatment.24 h before and after treatment,the blood samples from the vein of tail root of the rats in 4 groups were collected,and the serum expression levels of AchR-Ab and IFN-γ were detected with ELISA method.Results Compared with model control group and before treatment, the serum AchR-Ab and IFN-γ expression levels of the EAMG rats　in acupuncture treatment and drug control group after treatment  were significantly decreased (P<0.05 or P<0.01);compared with blank control group, the serum AchR-Ab and IFN-γ expression levels of the rats in model control group were significantly increased(P<0.01);but there was no significant difference between acupuncture treatment group and control group after treatment (P>0.05).Conclusion “Warming yang and Nourishing qi” acupuncture treatment can reduce the serum AchR-Ab and IFN-γ expression levels of the  EAMG rats to treat MG.

Abstract:Objective To study the influence of nitric oxide synthase(NOS) inhibitor L-aminoguanidine (L-AG) in the inflammatory factor levels in  degenerate intervertebral disc cells in vitro,and to explore its mechanism.Methods After treatment with L-AG(0,0.5%,1.0%,1.5%) for 24 h,the levels of NO and the activities of NOS were detected with spectrophotometric method and the expression levels of IL-1,IL-6 and TNF-α were determined by Western blotting.Results The activities of NOS in degenerate intervertebral disc cells in 0.5%,1.0%,1.5% L-AG groups were 6.1,5.5,and 4.2 U•mL^-1;the NO levels were 　 128.3,116.5, and 97.7  μmol•L-1,respectively,which were significantIy lower than those in OL-AG group(control group)（P<0.05）.Compared with control group，the expression levels of IL
^-1,IL-6 and TNF-α in  0.5%，1.0%，and 1.5%  L-AG  groups were also significantly reduced（P<0. 01-0.05）.Conclusion L-AG  can reduce the  inflammatory reaction of   degenerate intervertebral disc cells,which may be related to inhibition of NOS.

To observe the effect of the eukaryotic translation initiation factor 4E (eIF4E) targeted small interfering RNA (siRNA) on the eIF4E expression in human laryngeal Hep-2 cells,and to clarify the induction effect on apoptosis of Hep-2 cells and its mechanism.Methods The siRNA-eIF4E was transfected into human laryngeal carcinoma Hep-2 cells (siRNA-eIF4E group) with LipofectamineTM 2000,at the same time blank control group and empty plasmid group were set up.The inhibitory rate of cell proliferation in each group was detected by MTT.The changes of mitochondrial membrane potentials of the cells were observed by Rhodamine dyes.Apoptosis was detected by TUNUL staining,and the transcription and protein expression levels  of apoptosis related genes were analyzed by RT-PCR and Western blotting method.Results Compared with blank control group and empty plasmid group,the transcription and protein expression levels of eIF4E in Hep-2 cells in siRNA-eIF4E group were significantly decreased(P<0.01),the survival rate of Hep-2 cells was  decreased (P<0.05),the mitochondrial membrane potential of the cells was decreased (P<0.05),the apoptotic rate was increased (P<0.05),and the expression levels of apoptosis related genes Bim,Bid and Caspase3 were significantly up-regulated (P<0.05).Conclusion siRNA-eIF4E can inhibit the proliferation of human laryngeal carcinoma  Hep-2 cells in vitro,and its mechanism may be related to the activation of mitochondrial apoptosis pathway.

To study the influence of protein kinase Wee1B in the early development of mouse embryos by the means of specific Wee1B shRNA.Methods The mouse one-cell-stage embryos were collected after superovulation,and microinjected with TE buffer,control shRNA or Wee1B shRNA respectively,then the specificity of Wee1B shRNA was examined by the methods of RT-PCR and Western blotting.After RNA interference,the morphological changes and cleavage rate of mouse embryos were observed under fluorescence microscope,and the Wee1B activity of mouse embryos was detected by autoradiography.
Results Compared with TE buffer and Control  shRNA groups,the expression levels of Wee1B mRNA and protein  in Wee1B shRNA group were significantly reduced,and the activity of Wee1B was also decreased.The cleavage rates of mouse one-cell-stage embryos in non-injection,TE buffer,Control shRNA groups 33 h after hCG injection were 71.91 %±1.74%,72.90 %±2.25%, and 72.28 % ±2.06%,and there was no obvious difference among the three groups(P>0.05); the cleavage rate in Wee1B shRNA group  was  90.00%±0.77% at 33 h,which was markedly higher than those of the controls (P<0.01).Conclusion Wee1B shRNA can promote the  onset of mitosis of mouse one-cell-stage embryos,suggestting that Wee1B negatively regulates mitosis during early embryogenesis in mouse one-cell-stage embryos.

To explore the effects of isoflurane on autophagy and apoptosis  of rat PC12 cells induced by Aβ25-35,and to elucidate the possible mechanisms.Methods PC12 cells were randomly divided into 4 groups (n=6).Control group:the PC12 cells were administered with normal medium;10   μmol•L-1 Aβ25-35  group(Aβ group):the PC12 cells were administered with 10   μmol•L-1 Aβ25-35;2% isoflurane group(Iso group):the PC12 cells were administered with 2% isoflurane;2% isoflurane and 10   μmol•L-1 Aβ25-35 group(Iso+Aβ group):the PC12 cells were administered with 2% isoflurane +10   μmol•L-1  Aβ25-35.The autophagosomes of PC12 cells in various groups  were examined by transmission electron microscope,and the expressions of autophagy associated protein LC3-Ⅱ,p62 and autophagic signal gene mTOR,Beclin-1 were detected by Western blotting method 6 h after   administration;the  cell viability was determined by MTT assay,and  Western blotting method was performed to observe the expressions of apoptosis associated protein Bcl-2 and caspase-3 24 h after   administration.Results Compared with control group,great amounts of autophagosomes were presented in PC12 cells in Aβ group,the  expressions of LC3-Ⅱ,Beclin-1 and caspase-3 were significantly increased (P<0.05),and the  expressions of p62,p-mTOR and Bcl-2 were significantly decreased as well as  the  survival rate (P<0.05);however,autophagosomes failed to be presented in PC12 cells in Iso group,the  expressions of LC3-Ⅱ,Beclin-1 and Bcl-2 as well as cell survival rate were significantly decreased (P<0.05),and the expression of p62,p-mTOR and caspase-3 were significantly increased(P<0.05).Compared with Aβ group,only a few autophagosomes were found in PC12 cells in Iso+Aβ  group,the  expressions of LC3-Ⅱ and Beclin-1 as well as the  survival rate were significantly decreased (P<0.05),and the expressions of p62,p-mTOR and caspase-3 were significantly increased(P<0.05),but no changes occurred with the expression of Bcl-2 (P>0.05).Conclusion Isoflurane can inhibit autophagy  of rat PC12 cells induced by Aβ25-35 and aggravate the expressions of apoptosis rrelated proteins  through activation of autophagic signal
 gene mTOR and inhibition of Beclin-1.

Abstract:Objective  To observe the influence of fluoride in the pathological changes of incisor and the changes of physiological indicators of the rats with type 1 diabetes mellitus,and to clarify the effect of fluoride on process of diabetes mellitus. Methods 54 male Wistar rats were divided into control group (n=18),10 mg/kg/d F－group(10 mg　F－) (n=18)and 20 mg/kg/d F－group(20 mg　F－) (n=18).Fluoride was fed by gavage,1 month later 12 rats in each group were selected and treated with single intraperitoneal injection of 50 mg?kg-1 STZ to induce type 1 diabetes mellitus (used as STZ control group,10 mg　F－+STZ group,and 20 mg　F－+STZ group), then  the remaining 6 rats were fed with fluoride for 4 weeks(used as normal control group,10 mg　F－ group,and 20 mg　F－ group). In the experimental period,the changes of blood glucose were measured by glucose meter weekly,and the photograph was taken to follow teeth progressive changes. Before the end of experiment,intraperitoneal injection of insulin was performed to test the insulin sensitivity of the rats by insulin tolerance test(ITT).Results The rat dental face in 10 mgF－and 20 mg F－ groups gradually shifted from yellow to pale and turned to rough,uneven coloring,part of the teeth with white opaque changes,showing dental fluorosis alteration.Compared with normal control group,the rats in simple fluoride poisoning group had no change in weight,but after intraperitoneal injection of STZ,the weights of the rats in three groups showed downward trend,especially in 20 mg F－group,but there was no significant difference (P>0.05);after STZ induction the blood glucose levels were increased significantly (P<0.05) compared with normal control group;and the blood glucose level in 20 mg F－+STZ group had obviously change,but  there was no significant difference (P>0.05) compared with STZ control group.The ITT results showed that the glucose levels in 20 mg F－ group were significantly higher than those in normal  control group (P<0.05) at 0.5,1 and 2 h,while the glucose level in 10 mg F－group was elevated,but the difference was not statistically significant (P> 0.05).Conclusion High dose of fluoride can cause short-term dental fluorosis,aggravate the diabetes-induced body weight loss and blood sugar increasing,and decrease the insulin sensitivity in rats.

Abstract:Objective  To observe the effect of sodium aescinate(SA) on p38MAPK pathway in rats with intestinal ischemia reperfusion(I/R) injury, and to charify the protective mechanism of SA. Methods Twenty-four SD rats were randomly divided into three groups with eight rats in each:control group,intestinal I/R group,SA group.In control group,superior mesenteric artery (SAM)  was separated but not clamped;in I/R and SA groups,SMA was separated and clamped for 60 min.Each rat was given different treatment before ischemia,before reperfusion and 30 min after reperfusion respectively.In control and I/R groups,saline(5 mL/kg) was injected through tail;in SA group,SA(0.9 mg/kg) was injected through tail. The levels of TNF-α,IL-6 in plasma and the activities of myeloperoxidase(MPO) and diamine oxidase(DAO),malondialdehyde(MDA) levels in plasma and intestinal tissue were measured 60 min after reperfusion,as well as the expression levels of p38MAPK and Bax proteins in intestinal tissue were examined using immunohistochemical method.Results Compared with control group,the activities of MPO,DAO and the levels of TNF-α,IL-6,MDA in plasma were significantly increased;and the activity of MPO and the  MDA levels in intestinal tissue were also significantly increased,and while the DAO activity in intestinal tissue was significantly decreased in I/R group(all P<0.01).Compared with I/R group,the activities of MPO and DAO and the levels of MDA,TNF-α,IL-6 in plasma were significantly decreased;the MPO  activity and MDA level in intestinal tissue were also significantly decreased,and while the DAO activity in intestinal tissue was significantly increased in SA group(P<0.01).The immunohistochemistry results indicated that compared with control group,the protein expression levels of p38MAPK and Bax were significantly increased in I/R and SA groups(P<0.01);compared with I/R group,the p38MAPK and Bax protein expression levels were significantly decreased in SA group(P<0.01).The correlation analysis indicated that the MPO activity,the MDA level  and the expression levels of Bax protein in intestinal tissue  and the TNF-α,IL-6 levels  in plasma were positively correlated with the expression level of p38MAPK protein (r=0.797,0.702,0.712,0.695 and 0.715，all P<0.01).Conclusion SA can protect the intestinal I/R injury,which may be mediated by reducing the release of oxygen free radicals and cytokine,inhibiting activation of neutrophils,down-regulating p38MAPK gene protein expression to inhibit intestinal tissue apoptosis.

Abstract:Objective  To investigate the intervene effect of JieDu TongLuo BaoShen capsule (BaoShen capsule) on the  expressions of  adiponectin protein and monocyte chemoattractant protein-1 (MCP-1) in kidney tissue of  type 2 diabetic rats,and to clarify the renoprotective effect of BaoShen capsule and its mechanism.Methods Sixty-five male Wistar rats were divided randomly into normal control group (control,10 rats,fed standard rat chow),high-fat diet feeding group(High-fat control,10 rats,fed high-fat diet) and diabetic group(45 rats,fed high-fat diet).Four weeks later,type 2 diabetes was induced in the  rats in diabetic group by intraperitoneal injection of a low dose of streptozotocin.After modeling they were again randomly divided into untreated diabetic model group(DM,15 rats),BaoShen capsule(1.4 g/kg) treatment  group (BaoShen capsule,15 rats) and rosiglitazone combined with benazepril treatment  group (Positive control,15 rats),and treated for 12 successive weeks while continuing on the high-fat diet.Twelve weeks later,the blood glucose (BG),serum creatinine (Scr) and blood urea nitrogen(BUN) levels,urinary microalbumin excretion rate (UAE) were measured.The protein expression of  adiponectin in kidney tissue was determined using immunohistochemistry.Results Compared with control group,the  blood glucose,serum Scr and BUN levels,and UAE of the  rats in BaoShen capsule treatment group were significantly decreased(P<0.01);the serum adiponectin level  and the protein expression of  adiponectin in kidney tissue of the rats in DM group were markedly reduced   compared with control group(P<0.01),and the protein expression of  MCP-1 in kidney tissue was obviously elevated (P<0.01). Compared with DM group,the serum adiponectin level  and protein expression level of  adiponectin in kidney tissue in Baoshen capsule group were increased by 1.5 and 3 folds (P<0.01),while the MCP-1 protein expression level in kidney tissue was decreased by 55% (P<0.01).Correlation analysis indicated that the  serum adiponectin levels were negatively correlated with UAE(r=－0.466,P<0.01),Scr (r=－0.563,P<0.01),BUN (r=－0.512,P<0.01) levels and  MCP-1 protein expression level in kidney tissue (r=0.526,P<0.01).Conclusion BaoShen capsule treatment can increase the serum adiponectin level,up-regulates the adiponectin level in kidney tissue and and down-regulate the MCP-1 protein expression level in kidney tissue in type 2 diabetic rats,that is linked to renoprotective capacity of BaoShen capsule.

Abstract:Objective To observe the intervention effect of agaricus blazei polysaccharide (ABP) on ethology,mitochondrial membrane potential and apoptosis of hippocampal neuron in mice with brain aging induced by D-gal and to explore its effect and possible mechanism of improving the learning and memory ability of brain aging mice.Methods 40 Kunming mice were divided into four groups with 10 in each group:control group,aging model group,low dose ABP group and high dose ABP group.Morris water maze and avoiding dark instrument were used to detect the learning and memory ability of aging mice induced by D-gal while flow cytometry was used to analyze the changes of mitochondrial membrane potential and apoptosis.Results In the test of ethology,compared with model group,the percentages of distance in the central area of Morris water maze were obviously increased in ABP groups (P<0.05 or P<0.01),and the distances in the fourth quadrant were obviously decreased (P<0.01).Compared with model group,the mitochondrial membrane potential of hippocampal neuron in ABP groups were increased (P<0.01) and the apoptotic rates were decreased significantly (P<0.05 or P<0.01).Conclusion ABP can strengthen the learning and memory ability of aging mice and the mechanism is related to the increasing of  mitochondrial membrane potential of hippocampal neuron and the inhibition of apoptosis.

〗Abstract:Objective To investigate the influence of α-synuclein (SNCA) A30P point mutation in the neurogenesis in the olfactory bulb (OB) of adult mice，and to charify its effect in the pathogenesis of degenerative nervous diseases.Methods Normal C57BL mice,endogenous SNCA knockout mice (SNCA-/-) and SNCA knockout + human SNCA A30P knockin mice (A30P SNCA-/-) were used as experimental animals and were divided into C57BL,SNCA-/-, and A30P SNCA-/- groups.The coronal frozen sections of OB were made and the  morphological changes of  OB were  examined.Fluorescence immunohistochemistry of phosphohiston 3 (PH3),doublecortin (Dcx) and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) staining were carried out to analyze the number of  PH3+ cells,the fluorescence intensity in Dcx+ neuroblasts and the cell densities after DAPI staining in OB.Results The cell densities in glomerular layer (GL) of OB of the mice in SNCA-/- and A30P SNCA-/- groups were decreased slightly.A slight deformation of OB morphology also occurred in A30P SNCA-/- group and the number of PH3+ cells in A30P SNCA-/- group  was decreased significantly compared with C57BL and SNCA-/- groups (P<0.05);the fluorescence intensities of Dcx+ neuroblasts in OB in SNCA-/- and A30P SNCA-/- groups were increased slightly and no evident changes of the cell densities in OB after DAPI staining was observed.Conclusion Human SNCA A30P point mutation can decrease the neuron proliferation in OB of the adult mouse brain,which suggests it play a certain inhibitory effect on the neurogenesis in adult OB.

Abstract:Objective To investigate the inhibitory effect of antisense oligonucleotides (ASON) of telomerase on the growth of bladder cancer in vivo of nude mice and to confirm its therapeutic effect on bladder cancer.Methods Bladder carcinoma EJ cells cultured with medium and thiophosphoric acid-modfied ASON PS-ASON were inoculated into BALB/c mice as control group(n=24) and PS-ASON treatment group(n=12).The rates  of tumor genesis were observed in two groups.The mice in control group were divided into three groups(n=8)  depending on the different medicines fed to the mice as soon as 20 mm tumor formed.The mice in blank control group, PS-ASON  injection  group and chemotherapy medicine group were injected with saine,PS-ASON,and pharmorubicin,respectively.The tumor growth,weight of nude mice,tumor volumes and the survival rate were measured and observed.Then  the morphological changes of tumor tissue were observed  under light microscope.Results The rate of tumorgenesis of the mice in PS-ASON treatment group was lower than that in control group (P<0.01),the time of tumorgenesis in PS-ASON treatment group was more than that in control group (P<0.01) and the weight of the mice in treatment group was heavier than that in control group (P<0.01).After the mice were treated for 12 d,the tumor volumes of the mice in PS-ASON injection group and pharmorubicin  group were smaller than that in blank control group (P<0.01),but there was no significant difference in tumor volume between PS-ASON injection  group and pharmorubicin  group(P>0.05).The weight of the mice in blank control group lost obviously;the weight of the mice in PS-ASON injection group was bigger than that in blank control group(P<0.01) 16 d after inoculation.The weight of the mice in pharmorubicin  group were lower than that in PS-ASON injection  group(P<0.01) 8 d after inoculation;but there was no significant difference in the weight of the mice between blank control group and pharmorubicin  group(P>0.05).Necrosis and apoptosis of incubating tumor cells were observed in  PS-ASON injection  group and pharmorubicin group under light microscope.The survival rate of the nude mice in PS-ASON  group was higher than those in blank  group (P<0.001) and pharmorubicin  group(P<0.01).Conclusion ASON of telomerase inhibit the  growth of bladder cancer significantly in vivo.Its therapeutic efficacy is similar to the chemotherapy medicine,and it can be used for the further research on therapy for bladder cancer.

Abstract:Objective To investigate the influence of Curcumae decoction in p53 and caspase-3 expressions in hepatocytes of the mice with acute hepatic injury induced with carbon tetrachloride (CCl4),and to clarify the mechanisms of protective effect of Curcumae decoction on acute hepatic injury. Methods 60 ICR mice were randomly divided into blank  control group (saline for equal volume),model group (saline for equal volume),positive drug group (diammonium glycyrrhizinate),low dose of  Curcumae group (2.5 g/kg),middle dose of Curcumae group (5.0 g/kg) and high dose of  Curcumae group (10.0 g/kg).The acute hepatic injury mouse models were established with CCl4 after being given drugs for 7 d.The liver indexes were calculated,and the activities of alanine aminotransferase (ALT) in serum were detected and the pathological changes of liver tissue were observed.The gene transcription level of p53 in hepatocytes was examined by RT-PCR and the expression of caspase-3 protein in liver tissue was detected by immunohistochemistry. Results Compared with blank  control group,the liver index of the mice in model group was significantly increased (P<0.05);compared with model group,the liver indexes of the mice in different Curcumae groups were decreased (P<0.05). Compared with blank control group,the serum ALT activity of the mice in model group was increased (P<0.01),and the ALT activities of mice in positive drug group,high and middle doses of Curcuame groups were clearly decreased (P<0.05). The livers of the mice in blank  control group were red,soft and elastic,the livers in model group were swell,a little yellow,and having some filemot spalash.Microscopically,the liver tissue in blank control group was in gear，there were a lot of hepatocytes swell,inflammatory infiltration,balloon liked denaturalization and hepatocytes death of the mice in model group,but the lesions became much slighter in Curcumae groups.The gene transcription level of p53 in model group was increased compared with blank control group (P<0.05),and the gene transcription level of p53 in high dose of  Curcumae group was significantly decreased (P<0.05).The caspase-3 protein expressions in liver tissue of the mice in Curcumae groups were obviously decreased. Conclusion Curcumae can protect the liver from acute hepatic injury induced with CCl4 in the mice,its mechanism may be related to reducing  the gene transcription level of p53 and the expression of caspase-3 and prevent the hepatocytes from apoptosis.

Abstract:Objective  To explore the application of c7c phage-display peptide library in screening the short peptides which bind specifically to high metastatic potential hepatocellular carcinoma cells HCCLM3,and to lay foundation for research on the molecular mechanism of hepatoma metastasis and detection of the markers of hepatoma metastasis.Methods The human high metastatic potential hepatocellular carcinoma cells HCCLM3 were used as the target cells and the human low metastatic potential hepatocellular carcinoma cells SMMC7721 as the absorber cells for subtraction biopanning from c7c phage-display peptide library.The affinity and specificity of the positive phage clones were identified by ELISA and immunocytochemistry,and the amino acid sequences were deduced by DNA sequencing.Results  After 4 rounds of screening,the phages which can bind to the HCCLM3 cells were enriched obviously (P<0.05). 6 positive phage clones (C4,C6,C7,C10,C11 and C17) with high affinity were identified by ELISA. After cell immunocytochemistry,1 peptide namely C7 with higher specificity to HCCLM3 cells compared with negative control was found (P<0.05) and no conserved motif was found in these peptides.Conclusion The peptides specifically binding to HCCLM3 cells are screened and identified from c7c phage-display peptide library.

Abstract:Objective To explore the protective effect of Schisandra polysaccharide (SCP) on acute chemical liver injury induced by carbon tetrachloride (CCl4) in mice and to reveal its mechanism.Methods 50 mice were randomly divided into normal control group,model group,low dose of SCP group (SCP,50 mg/kg),high dose of SCP group (SCP,100 mg/kg) and positive control group (bifendate,150 mg/kg).All the mice were intragastrically administrated the corresponding agents respectively for 15 d.1 h after the last administration,10% CCl4 (2.0 mL/kg) was given to mice in all groups except control group in a single intraperitoneal injection to establish the acute liver injury mouse models.6 h later,the mice were sacrificed,and the liver index,alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities,total bilirubin (TBIL) level in serum were detected, and the malondialdehyde (MDA) and glutathione (GSH) levels,and nitric oxide synthase (NOS) activity in liver tissue were tested.HE staining was performed for observing the pathological changes of the liver tissue.Results Compared with normal control group,the liver quality,liver index,ALT,AST levels and TBIL level in serum,the MDA level and NOS activity in liver tissue in model group were all increased significantly (P<0.01),and the GSH level  in liver tissue was decreased (P<0.01).Compared with model group,the liver quality and liver index of the  mice in high dose of SCP group were decreased (P<0.05);the ALT and AST activities in serum,and the MDA level  in liver tissue in low dose of  SCP,high dose of SCP and positive control groups were decreased significantly (P<0.05 or P<0.01),while the GSH levels  in liver tissue were increased (P<0.05);both the serum TBIL level and NOS activity in liver tissue in high dose of SCP group were reduced (P<0.05 or P<0.01).Moreover,the HE staining results showed that the size of liver cells was increased and the cytoplasm was stained lightly,and eosinophilic degeneration could be also seen in some individual liver cells in model group.The morphological damage of liver cells and the eosinophilic degeneration were significantly improved in SCP group.Conclusion SCP has a significantly protective effect on CCl4-induced acute liver injury,which might be related to the anti-oxidation.

Abstract:Objective To study the expression levels of Sox9 and collagen Ⅱ in condylar chondrocytes after treated with different modes of mandibular advancement in rats,and to clarify the effects of different modes of mandibular advancement on the expressions of Sox9 and collagen Ⅱ as well as formation of cartilage in condyle.Methods  Twenty-eight male 35-day-old Wistar rats were randomly divided into dynamic,static,functional and control groups(n=7),and received different modes of mandibular advancement according to their groups.2 weeks 　after the experiment,temporomandibular joint sections were prepared for immunohistochemical staining,and the expression levels of Sox9 and collagen Ⅱ in condylar chondrocytes were semi-quantitatively evaluated.Results The number of Sox9 positive cells in control group,dynamic group and functional group was significantly higher than that in static group(P<0.01),the Sox9 positive expression areas in dynamic group and control group were statistically larger than those in static group and functional group(P<0.01),the positive expression area of collagen Ⅱ in functional group was significantly higher than those in the other three groups(P<0.01),and the positive  expression area of collagen Ⅱ in static group was higher than that in control group(P<0.05).Conclusion The modes and duration of mandibular advancement play a key role in regulating the expressions of Sox9 and collagen Ⅱ in condylar chondrocytes.

Abstract:Objective To compare the effects of five different titanium surface characteristics of implants on gingival soft tissue interface and to find a proper implant neck surface characteristic for the preferable adhesion of the gingival soft tissue.Methods The  titanium implants were  processed by electrochemical methods to form four different surfaces,namely the Nano morphology of acid etching group (AE group),the Micron morphology of electrochemical etching group 1 and 2 (ECE1 group and ECE2 group),the micro or Nano morphology of electrochemical etching and acid etching group(ECA group);and the smooth morphology in smooth group (S group) was used as control.These five kinds of implants were immediately implanted in the mandible of 6 experimental dogs in a certain order.The dogs were sacrificed respectively after 20 d and 3 months,and then the specimens were manufactured and scanning electron microscope was used to observe the bonding interface of the gingival with the implant surface.Results The gingival epithelium bonded with the implant surface closely.No significant difference was found among all groups;after removal of soft tissue,there was no epithelial tissue remaining on implant surfaces.The collagen fibers in S group adhered and paralleled with the implant surface,and most of the collagen fibers dehisced and separated from implant surface;and few residual collagen fibers remained on implant surface.The collagen fibers in other groups were vertical to the implant surfaces.The interface of gingival tissue and implant surface formed complex micro-Nano mosaic structures,and the gingival tissue dehisced less from implant surfaces compared with S group;the collagen fibers remaining on the implant surface  in ECE2 group and  ECA  group were more than those in AE group and ECE1 group.Conclusion The micron and micron - nanometer surface can significantly  improve the adhesion of the gingival soft tissue on the implant with the methods of electrochemistry.

Abstract:Objective To compare the changes of drug-susceptibility in antibiotic and antimicrobial Chinese herbs before and after long time passaged,and to detect the genetic mutation of some resistance genes. Methods The Escherichia coli (E.coli) ATCC25922 was cultured in the M-H broth culture including Levofloxacin or Yinhuamiyanling by 200 passages.The  minimum inhibitory concentration (MIC) of E.coli ATCC25922 of these two medinices was detected and compared with the MIC of untreated E.coli ATCC25922.The bacterial of  the genes encoding gyrase(gyrA) and topoisomerase Ⅳ were amplified by PCR.Results The MIC of the bacterial cultured in the M-H broth culture including Levofloxacin  was increased compared with the MIC of untreated E.coli.There was no significant difference in the MIC of the bacteria cultured in the M-H broth culture including Yinhuamiyanling between before and after treatment(P>0.05).Several mutations were identified in gene of parC.The adenine(A) at the site 381 of parC was converted to thymine(T),and a thymine(T) was inserted in at the site 386.All isolates harbored parC amino acid subctitutions at position 127(Lys127-Asn).The gene gyrA had no mutation.Conclusion Levofloxacin is easier than Yinhuamiyanling to produce the resistance of E.coli ATCC25922,and the mutation of parC gene may be one of reseason of bacterial resistance.

Objective  To investigate the antimicrobial effects of disposable ofloxa
cin eye drops chosen randomly from the market on the bacteria and fungus,and to
 expound that the eye drops without preservatives still have the proper bacterio
static effect.Methods The minimum inhibitory concentration (MIC) of Staphyloco
ccus aureus，Escherichia coli and Pseudomonas aeruginosa　were determined bybroth microdilution method specified in Chinese Phrmcopoei.
The bacterial colony of Candida albicans and Aspergillus niger in different time points（0，7，14 and 28 d）after treated by ofloxacin were recorded.Non-disposable ofloxacin eye drops was used as positive control group.Results 7,14,28 d after treatment of disposable ofloxacin eye drops,the fungus number of Aspergillus was not increased and the growth of Candida albicans was almost inhibited.The disposable ofloxacin eye drops also had significantly
 inhibitory effect on Staphylococcus aureus，Escherichia coli and Pseudomonas aeruginosa,and the MIC was 5.9 mg?L-1.The antimicrobial efficacy of disp
osable ofloxacin eye drops was similar to that of non-disposable ofloxacin eye drops.Conclusion The antimicrobial effect of disposable Ofloxacin eye drops selected from the market all meet the requirements of ophthalmic preparations in Chinese Pharmacopeia.The bacteriostatic agent wit
hout preservatives also has proper bacteriostatic effect.
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Objective To study the antibacterial activities of 95% ethanol extract
of Yinhuamiyanling Tablets in Eschericha coli  (E. coli) standard strains and clinical drug-resistant strains in vitro,and to clarify the antibacterial mechanism.
Methods Cylinder-plate method was used to test the antibacterial activities of Yinhuamiyanling Tablets of 95% ethanol extract on E.coli standard s
train ATCC25922 and 18 strains of urinary tract infection clinical E.coli producing extended spectrum β-lactamases (ESBLs),and the minimum bactericidal concentration (MBC) was determined.The bacterial ultrastructural changes were observed by scanning and transmission electron microscope before and after the drug application.Results The MBC50 and MBC90 of 95% ethanol extract of Yinhuamiyanling Tablets in  E.coli standard strain ATCC25922 and clinical E.coli producing ESBLs were 100 g/L^-1 and 150-250 g/L^-1.The E.coli cells showed surface collapse,depression and thorn-like protuberances at both ends,severely deformed under  scanning electron microscope after treated with 95% ethanol extract of Yinhuamiyanling Tablets;negative staining revealed that the bacterial refraction deteriorated under transmission electron microscope,bacteria deformation appeared to fold,and part of
the cell wall were depression;the negative staining  transmission electron microscope results  showed  bacteria shrinkage,swelling,the cell wall blurred or even
 missed,and the cytoplasm was pyknotic and disintegration to small vacuoles.Conclusion 95% ethanol extract of Yinhuamiyanling Tablets  has a certain degree of inhibitory effect on E.coli standard strain ATCC25922 and E.coli producing ESBLs,and its mechanism may be related to its interference or disruption of the E. coli
cell wall synthesis or destruction of large molecular proteins,which results in  the ultrastructural changes of E.coli.
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Objective To prepare a new hand-washing agent with heterpolyoxometalates and to study its anti-A-influenza virus effect and toxicity.
Methods The anti-virus effects of the two compounds K10Na［(VO)3(SbW9O33)2］?26H2O(PM-1001) and K15［Pr(BW11O39)2］?28H2O(Pr-B)　were detected on MDCK cells.Three levels  of heterpolyoxometalates(high,middle,and low doses)were chosen to add into hand-washing agents; the anti-A-influenza virus effect
 
test was performed according to the American Society for Testing and Materials(ASTM) standard assessment protocols.In this test,10 volunteers were randomly divided into two groups for testing hand-washing agents with PM-1001 and Pr-B respectively.Every volunteer’s ten fingers were divided into 5 groups randomly.After all the finger taps were contaminated with 0.02　mL of 107.5 tissue culture infectious dose (TCID50)/0.1　mL live human influenza A virus, the H1N1 titers  were assessed with MDCK cell culture method after  contaminating,air drying and treatment with different hand-washing agents.The
concentrations and  viabilities of H1N1 on hands  30 and 60 min after hand drying  were also assessed.The hand-washing agent with  high dose of PM-1001 was chosen for toxicity assessment using Kunming mice and rabbits according to Disinfection Technical Specifications (2008).
Results The  therapeutic indexes  (TI) of PM-1001 and Pr-B   were  82.68 and 69.13.The H1N1 concentration in  control group was 104.33±0.66TCID50/0.1　mL
.The fingers in treatment groups had no viable H1N1 recovered.The virus concentrations in  treatment groups had significant differences compared with  control group(P<0.001).The H1N1 virus concentrations  in  30 and 60 min groups were 103.98 ±0.42TCID50/0.1　mL and 103.83±0.35TCID50/0.1 mL and had no significant differences compared with  control group(P>0.05).The LD50 in  mouse acute toxicity test  was >5 000 mg/kg^-1,the rabbit skin irritation index was 0.33,and the eye stimulation indexes of three rabbits were 0,0.67, and 0.33. Conclusion The new  hand-washing agents including PM-1001 and Pr-B are highly effective in reducing influenza A virus on human hands.It has no toxicity and no stimulation to skin and eyes of rabbits.
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Objective To investigate the relationship between angiotensinogen (AGT) gene M235T polymorphism and hypertrophic cardiomyopathy(HCM) and to provide e
vidence for prevention and treatment of HCM. Methods PubMed,Embase,OVID,Web of Science,Cochranelibrary,CNKI,WangFang Data and VIP were searched to identify the studies about the relationship between AGT M235T polymorphism and HCM.The studies that met the inclusion criteria were used for Meta-analysis with Stata 11.0 software.Results  Eight studies were included,which involved 887 cases and 1 407 controls. The Meta-analysis results indicated that there was no

significant association between AGT M235T polymorphism and HCM（T vs M：OR=1.17，95%CI=0.95－1.45；(TT/MT) vs MM：OR=1.12，95%CI=0.87－1.45；TT vs (MM/MT)：OR=1.21，95%CI=1.00－1.45；MT vs MM：OR=1.07，95%CI=0.82－1.41；TT vs MM ：OR=1.25，95%CI=0.79－1.99. In subgroup analysis,the significant difference of association between AGT M235T polymorphism and HCM existed in Asian and sporadic hypertrophic cardiomyopathy (SHCM),but no significant difference was found in European and familial hypertrophic cardiomyopathy (FHCM).Conclusion There is no association between HCM and AGT M235T polymorphism in populations,but such a relationship exists in Asians and SHCM.

Objective To investigate the association between angiotensin Ⅱ type 1 receptor (AT1R) gene A1166C polymorphism and essential hypertension (
EH) in Uygur nationality and to clarify the action mechanism of AT1R gene A1166C in the occ
urrence of EH.Methods 636 cases of Uyghur population blood samples including 167 cases of EH group and 469 cases of control group were collected.
Then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP )method was adopted to examine the AT1R gene A1166C polymorphism.The distribution of AA,AC and CC genotypes and the different frequencies of the points A,C in EH and  control groups were analyzed.
Results The AC+CC genotype and C allele frequency in EH group were significantly higher than those in control group,the difference was statistically significant（χ2=10.179 ，P=0.001）;the  risk of EH of the people with AC+CC genotypes was 1.6（OR=1.612，95%CL:1.191-2.183）times than that of the people with AA genotype.Conclusion There might be some relationship between AT1R gene A1166C polymorphism and the EH in Uighur population.

Objective  To study the expressions of Musashi1 and sex determining re
gion Y-box 2 (Sox2) proteins in lung cancer,benign lesion lung and normal cancer
-adjacent lung tissues,and to explore the clinical significances of Musashi1 and Sox2 proteins in the carcinogenesis and progression of lung cancer.
Methods The expressions of Musashi1 and Sox2 proteins were detected by immunohistochemistry in 50 cases of lung cancer tissue,32 cases of benign lesion lung tissue and 18 cases of normal cancer-adjacent lung tissue.The relationships between the expressions of Musashi1 and Sox2 proteins and clinical pathological parameters were analyzed.Results  The positive rates of Musashi1 expression in lung cancer tissue,benign lesion lung tissue and normal cancer-adjacent lung tissue were 50% (25/50),0 (0/32),and 0 (0/18).The expression rate of Musashi1 in lung cancer tissue was significantly higher than those in benign lesion lung tissue and normal cancer-adjacent lung tissue (P<0.001).The expression of Musashi1 was significantly correlated with the histological types and differentiation of lung cancer(P<0.05),but was not correlated with the age,gender,TNM stage and lymph node metastasis (P>0.05).The positive rates of Sox2 moderate to strong expression in lung cancer tissue,benign lesion lung tissue and normal cancer-adjacent lung tissue were 64% (32/50),9.4% (3/32),and 0 (0/18). The expression rate of Sox2 in lung cancer tissue was significantly higher than those in benign lesion lung tissue and normal cancer-adjacent lung tissue (P<0.001).The expression of Sox2 was significantly correlated with the histological type of lung cancer(P<0.001),but was not correlated with the age,gender,TNM stage,lymph node metastasis and differentiation (P>0.05).The expression of Musashi1 in lung cancer tissue was positively correlated with the expression of Sox2 （r=0.598,P<0.001).Conclusion The up-regulation of Musashi1 and Sox2 in lung cancer tissue suggests that they may play crucial role in the process of carcinogenesis and progression of lung cancer.Musashi1 and Sox2 may be used as the potential diagnostic markers and therapeutic targets in lung cancer.
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Objective To explore the expression levels of IL-17,IL-23 and TGF-β
 in peripheral blood and bone marrow of the patients with aplastic anemia(AA) and its clinical significances,and to clarify the pathogenesis of AA.
Methods  50 cases of AA patients were randomly selected as experiment group and 30 cases of
healthy people at the same period were used as  control group;Enzyme-linked immunosorbent assay(ELISA) and real-time fluorescence quantitative were performed to detect the expression levels of IL-17,IL-23 and TGF-β proteins and mRNA in peripheral blood and bone marrow.
Results  Compared with control group,the protein and mRNA levels of IL-17 and IL-23 in peripheral blood and bone marrow of the patients in experiment group were significantly increased,and the differences between two groups were statistically significant(P<0.01),among them,the  elevated IL-17 was significant(P<0.001);the protein and mRNA levels of TGF-β in peripheral blood and bone marrow of the patients in experiment group were significantly
 decreased,and the differences between two groups  were also statistically significant (P<0.01).The protein and mRNA levels of  IL-17 and IL-23
 in peripheral blood and bone marrow of the patients in experiment group had  positive correlation (r=0.69,P=0.041；r=0.71，P<0.05),while the pr
otein  levels of  IL-17  and TGF-β had  negative correlation(r=－0.65,P<0.05);and the potein levels of IL-23 and TGF-β had  negative correlati
on(r=－0.72,P<0.05).Conclusion IL-17,IL-23 and TGF-β may participate in the occurrence and development of AA,and the combined dete
ction of the three indexes provides the new ideas for clinical treatment of AA.
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Objective  To detect the expressions of survivin,galectin-3(Gal-3) and thyroid peroxidase(TPO) in papillary thyroid microcarcinoma (PTMC) tissue and cancer
adjacent normal tissue,and to explore their clinical significances.Methods  The expressions of survivin,Gal-3 and TPO
 in 56 cases of PTMC tissue and adjacent normal tissue were measured by immunohistochemical (IHC) method;Chi squ
are test was used to compare the differense in the expression rates of survivin,Gal-3 and TPO. The relationships between the ex
pression rates of survivin,Gal-3 and TPO proteins and the clinicopathological parameters of PTMC were analyzed.
Results  The survivin and Gal-3 were strongly expressed but TPO was negatively expressed in PTMC tissue.The survivin and Gal-3
 were negative in adjacent normal tissue but TPO was expressed i  PTMC tissue.There were significant differences in the expression rates of survivin,Gal-3 and TPO between PTMC tissue and adjacent normal tissue (P<0.001).The expression rates of survivin,Gal-3 and TPO in PTMC tissue were positively related  to lymph node metastases (P<0.05, or P<0.01),and were negatively related to gender and age (P>0.05).   Conclusion The strong expressions of survivin and Gal-3 and
 the mild expression of TPO may be important in the development of PTMC, and the detection of survivin,Gal-3 and TPO protein expressions is helpful to jude
 benign and malignant PTMC.

Objective To explore the  expressions of osteopontin(OPN) in normal ovarian tissue,benign ovarian tumor tissue
 and epithelial ovarian carcinoma tissue,and to clarify the value of OPN as an indicator to  monitor the malignancy degree and evaluate the prognosis of  epithelial ovarian carcinoma. Methods The ovarian resection specimens of 100 cases were collected,including 20 cases of  normal ovarian tissue,20 cases of benign ovari
an tumor tissue and 60 cases  of epithelial ovarian carcinoma  tissue.Epithelial ovarian carcinoma including 30 cases of ovarian serous adenocarcinoma,20 case
s of ovarian mucinous cystadenocarcinoma and 10 cases of ovarian endometrioid carcinoma.According to the  differentiation degree,60 cases of epithlial ovarian carcinoma were  divided into 12 cases  well differentiated carcinoma (G1),16 cases of  moderately differentiated carcinoma (G2),
and 32 cases of poorly differentiated carcinoma (G3).According to the International Federation of Gynecology and Obstetrics (FIGO) 2000 primary ovarian cancer surgery - pathological staging criteria,60 cases of epithelial ovarian carcinoma including 30 cases of epithelial ovarian carcinoma at stage Ⅰ and  Ⅱ,and 30 cases of  epithelial ovarian carcinoma at stage Ⅲ and  Ⅳ.HE staining and immunohistochemical staining  were used to detect the OPN ex
pression.Results OPN  mainly expressed in the cytoplasm of cancer cells. The positive  expression rate  of OPN in 60 cases of epithelial ovarian carcinoma group was 75.0% which was  higher than those in normal ovarian group  and benign ovarian tumor group(P<0.01).With the increasing of clinical stage,the  positive expression rate of OPN was increased;the positive expression rate of OPN in poorly differentiated epithelial ovarian carcinoma group was  higher than that in well differentiated epithelial ovarian carcinoma group (P<0.01);the positive expression rates of OPN had no significant   differences  between different histological types of epithelial ovarian carcinoma (P> 0.05).Conclusion OPN may contribute to the occurrence, development,invasion, and metastasis of epithelial ovarian carcinoma.OPN is of great significance for the clinical assessment of  diagnosis and metastasis of epithelial ovarian carcinoma,and it can be used as
an important indicator to  monitor the malignancy degree and evaluate the prognosis of  epithelial ovarian carcinoma.

Objective To explore the changes of NK cells and  the expression levels of their activated and inhibitory receptors in peripheral blood of the patients
with systematic lupus erythematosus (SLE) before and after treatment of glucocorticoid combined with immunodepressant,and to clarify its action mechanism on SLE.Methods 26 patients with new onset SLE and 16 healthy controls were selected.The expression levels  of NK cells,their activated and inhibitory receptors i
n peripheral blood in two groups were detected before treatment, 4 and 12 weeks after treatment by flow cytometry.Results  Compared with control group,the
ratio of CD3－CD56+NK cells of the patients with SLE and the expression levels of KIR2DL3+,KIR3DL1+,NKG2A+ NK in SLE patients group before treatment were significantly decreased(P<0.05);the expression levels of NKG2C+，NKP30+，NKP46+, and the ratios of IFN-γ+ NK cells were significantly increased(P<0.001).Compared with before treatment,the ratios of CD3－CD56+NK cells in the patients with SLE 4 and 12 weeks after treatment were significantly inc
reased(P<0.05）;the expression levels of NKG2C+,NKP30+, and NKP46+in the patients with SLE 4 weeks after treatment were significantly decreased(P<0.05).4 weeks after treatment,the expression  levels of KIR2DL3+,KIR3DL1+,and NKG2A+  were significantly increased compared with before treatment(P<0.05).12
weeks after treatment,the expression  levels of KIR3DL1+,CD158a+,CD158b+, and NKG2A+ were significantly increased compared with before treatment(P<0.05),and the ratios of IFN-γ+CD３－ CD56+ NK cells in the patients with SLE 4 and 12 weeks after treatment were significantly decreased compared with before treatment(P<0.001).Conclusion The changes of the NK cells and its receptors may be involved in the pathogenesis of SLE.Glucocorticoid combined with immunodepressant may play the therapeutic effect  by regulating the expression of NK cells and their receptors.

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Objective To observe the sensitivity of t(17;19) acute lymphoblastic leukemia (ALL) cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) -mediated apoptosis and to discuss its possible mechanism and clinical significance.Methods 4 t(17;19)-ALL cell lines and a primary cell sample were used as experimental group and 28 other B-precursor ALL cell lines were used as control group.The cell surface expression of TRAIL receptors was detected by flow cytometry,and the inhibition of cell growth was measured by 3H-thymidine uptake assay after treated with rhsTRAIL.Early apoptosis was detected by flow cytometry using Annexin-V staining and the changes of apoptosis signaling pathway after incubation with rhsTRAIL were checked by immunoblotting assay. Results The cell surface expression of death receptor 4(DR4) in t(17;19)- ALL cells was significantly higher than that in other cell lines (P<0.05).The relative fluorescence intensity (RFI) of DR5 was higher than 1.4 while it was lower than 1.3 in other cell lines.The cell growth of t(17;19)-ALL was nearly 100% inhibited by rhsTRAIL at 100 μg/L,which was much higher than other cell lines (P<0.05 or P<0.01).The inhibition was blocked by RIK-2 (neutralizing antibody of TRAIL) and  z-VAD-fmk ( inhibitor of caspase).rhsTRAIL induced early apoptosis in t(17;19)-ALL cells and the apoptotic rate was much higher than that in negative control (P<0.05).Cleaved bands of caspase-8,Bid,caspase-3 and PARP were observed within2 h of incubation with rhsTRAIL. Conclusion t(17;19)-ALL cells are highly sensitive to TRAIL and eventually to graft versus leukemia (GVL).Allo-SCT should be performed in early stage to obtain long-term surviving in the patients with t(17;19)-ALL.

Abstract:Objective To investigate the effect of using the VivaSens desensitizier on tooth-whitening surface before bleaching with Beyond cold light and efficacy of bleaching and to clarify its application value in cold light bleaching. Methods 15 patients who deserved the treatment were chosen,in total of 299 teeth， their upper or lower dentitions were used as a sample.The upper dentitions (including anterior teeth and premolar teeth) or lower dentitions(including anterior teeth and premolar teeth ) from the same patient were randomly divided into experimental group(n=15,150 teeth，8 upper dentitions，7 lower dentitions) and control group(n=15,149 teeth，7 upper dentitions，8 lower dentitions).Before bleaching,the VivaSens desensitizer was applied on the buccal surfaces in experimental group,and a placebo gel was applied on the buccal surfaces in control group.The tooth hypersensitivities of the patients were measured by probe and cold air test before and after treatment with a 0-to-10 scale,and the postoperative 101 teeth（94%） in experimental group and all of teeth in control group experienced dental hypersensitivity right after bleaching(P<0.01).After 24 h,22 teeth（14.7%） in experimental group still had dental hypersensitivity,while 75 teeth（50.3%） in control group had,there was significant difference of incidence of dental hypersensitivity between two groups (P<0.01);after 48 h,the dental hypersensitivity in experimental group  disappeared,but 16 teeth（10.7%） in control group still had,there was significant difference of incidence of dental hypersensitivity between two groups (P<0.01).The dental hypersensitivities in experimental groups during different stages were lower than those in control group(P<0.05).The Vita Shade Guide results showed that  before and after bleaching,experimental and control group showed a change of 5 Shade Guide Units after treatment compared with before treatment;and there was no difference of bleaching effect between two groups(P>0.05).Conclusion VivaSens desensitizer could effectively reduce the incidence and intensity of postoperative hypersensitivity without affecting the bleaching effect.

Abstract:Objective To observe and evaluate the clinical effects of  mitck  mini anchor in treatmentfor acute closed starting ending point injury in lateral collateral ligament of interphalangeal joint.Methods 14 patients（15 fingers） with acute closed starting or ending point  injury in lateral collateral ligament of interphalangeal joint were treated by operation and anchor planted  in phalanx to rebuild its starting or ending points.Closed-plaster method was  used for 2 weeks after the operation before the protective function practice.Lateral stress test,ROM,tumentia,deformity,pain and X-ray were observed when the patients were rechecked.Results All the patients were followed up for  6 to 15 months,averaged for 10.7 months.The interphalangeal joint of the patients  was  stable and had no pain and tumentia,  and the ROM was 70°-110°,averaged for 91°.According to Dargan function evaluation standard, 12 fingers were excellent,2 fingers were good,1 finger was fair,and the fineness  rate was 93.33%.Conclusion The  method of reconstruction  of mitck mini anchor in treatment for acute closed starting or ending point  injury in lateral collateral ligament of interphalangeal joint   is easy  and  has satisfied efficacy.

Abstract:Objective To provide the standard morphological parameter values of hippocampal through linear measurement of the hippocampus and the temporal horn of the lateral ventricle in healthy subjects.Methods The areas,heights,and widths of the hippocampus and the temporal horn of the lateral ventricle were measured in 3 standard planes on magnetic resonance images(MRI) in 105 healthy subjects,and the comparisons between left side and right sides and between various gender groups were performed.Results The difference of the areas of the hippocampus between the two sides of brain had no significance (P>0.05).However,the differences of width(right side>left side,P<0.01) and heights(left side>right side, P<0.01)  of the hippocampus between the two  sides were significant.There were no significant differences of the widths,heights and areas of temporal horn of the lateral ventricle between the two sides(P>0.05). The difference of the measurement values of the hippocampus and the temporal horn between the male and the female had no significance (P>0.05).Conclusion There are differences of the width and height of the hippocampus in standard planes between the two sides.The linear measurement standard values of the hippocampus and the temporal horn of the lateral ventricle obtained in standard planes will help the clinical diagnosis in related diseases.

Objective To investigate the  blood pressure level and  the prevalence and distributional characteristics of hypertension among the adults in Jilin province and to provide evidence for making  preventive and controlling strategies against hypertension.Methods A total of 20 473 adults aged 18-79 years old from Jilin province were selected by multistage cluster randomly sampling.Medicial examination and face-to-face questioning survey were conducted to collect the information about hypertensive distribution.Blood pressure was examined by electronic blood pressure monitor.According to complex sampling plan and post-stratification,the resultant data were weighted to estimate the level of blood presure and the prevalence rate.Results  The weighted average systolic blood pressure and diastolic blood pressure were (128.1±20.2) mmHg and (78.4±11.6) mmHg,respectively. The weighted prevalence rate of hypertension was 30.5%(95%CI：29.7%-31.2%). The prevalence rate was increased with age. The prevalence rate of the males (34.4%，95%CI：33.3%-35.6%) was statistically higher than that of the females(26.3%，95%CI：25.3%-27.2%) (P<0.01).The prevalence rates  of hypertension in easten region (33.2%，95%CI：31.4%-35.1%) and western region (32.4%，95%CI：30.5%-34.3%) were higher than that in central region (29.0%，95%CI：28.1%-29.9%) (P<0.01).There was nosignificant difference between the prevalence of hypertension in urban area (30.4%，95%CI：29.3%-31.4%) and rural area (30.6%，95%CI：29.6%-31.7%) (P>0.05).Conclusion In 2012,the prevalence rate of hypertension among the  adult residents in Jilin province is higher than before.There are differences in the distributional characteristics of hypertension among variations by age,gender and geographic regions.Health system should prevent and control hypertension according to the epidemiological characteristics.

Objective To study the awareness,treatment and control rates of hypertension in the  adult residents in Jilin province,and to provide scientific basis for  prevention and control of hypertension.Methods A random cluster sampling survey was taken in   20 437 people from 9 cities and 32areas in Jilin province in 2012.A cross-sectional study with questionnaire interview and physical examination were carried out for every participant.After being weighted according to complex sampling scheme and post-stratification Rao-Scott test which based on sampling design was used to compare the awareness,treatment and control ratesof hypertension between  people with different demographic characteristics.Results After being weighted,the weighted rates of awareness,treatment and control of hypertension of the adult residents in Jilin province were 44.1%,41.3% and 7.9%,respectively. The weighted rates of awareness,treatment and control of hypertension  in the male subjects were lower than those in the female subjects(P<0.001).The rates of awareness and treatment of hypertesion  in people aged 18-24 years were 9.6% and 9.5%,respectively,which  were the lowest.There  weighted rates of awareness and treatment of hypertension  in the people with high  education level was lower than those in  the people with low education level (P<0.05).The weighted rates of awareness,treatment and control of hypertension  in the people with positive hypertension family history were higher than those in the people with  negative hypertension family history(P<0.001).The weighted rates of awareness and treatment  of hypertension  in  rural  areas were higher than those in urban areas(P<0.05)，but there was no significant difference in the control rate between rural and urban areas(P>0.05).Conclusion The weighted rates of awareness and treatment   of hypertension of the adult residents in Jilin province are lower than the average of country in 2004-2005,but the rate of control is higher than the average of country;because the rate of awareness of hypertension in young men is low,it is important to improve the prevention and control of hypertension education level among young men.

Objective To investigate the condition of neuroses in the patients with hypertension of Changchun city in Jilin province ,and to analyze its influencing factors and provide evidence for the prevention and treatment of the  patients with hypertension. Methods Stratified cluster sampling method was conducted in the local health service centers. 15 health service centers were chosen. 1 216 patients with hypertensionwere informed by the doctors and 999 participants who were interviewed face to face using the version 3.0 of the World Health Organization Composite International Diagnostic Interview（WHO-CIDI-3.0） successfully completed the interviews and diagnosed by International Statistical Classification of Diseases and Related Health Problems,tenth revision(ICD-10). Lifetime prevalence,12-month prevalence and 30 d prevalence were used to describe the different kinds of neuroses in the patients with hypertension. The  χ2 test was used tocompare the  distribution characteristics of 12-month prevalence of neuroses in the patients with hypertension between various groups. Logistic regression was used to analyze the  general demographic characteristics and psychosocial factors. Results The lifetime prevalence of neuroses in the patients with hypertension was 14.6%,the 12-month prevalence was 10.8%,and the 30 d prevalence was 7.9%.The Logistic analysis results showed that four influencing factors were found,age(60-69 years,≥70 years),negative life events,positive coping style,and life satisfaction and their OR（OR95%CI） values were 0.412（0.216,0.784）,0.308（0.139,0.683）,1.010（1.003,1.017）,0.670（0.454,0.989）,and 0.934（0.896,0.974）,respectively. Conclusion The prevalence of neuroses in the patients with hypertension is higher. Age is one of the influencing factors,and after 60 years old,the older the patient is,the lower prevalence of neuroses. Negative life events can increase the risk of neuroses in the patients with hypertension. Positive coping style and high life satisfaction can reduce the risk of neuroses in the patients with hypertension.

Objective  To probe into the major problems and propose the corresponding countermeasures through analyzing the flow direction of the hospital visit and medical expenditure of new rural cooperation medical scheme (NRCMS) in Jilin province.Methods The 2003-2012 data from the statistical information system of NRCMS in Jilin province  was used to conduct the statistic analysis.A cluster sampling method was used to investigate  NRCMS inpatient,about person-time,medical expenditure,hospitalization reimbursement and  rate per time in various hospitals.Results During ten years from 2003 to 2012,the rate of inpatients,medical expenditure and reimbursement to county-level or higher-level medical institutions increased year by year.The constituent ratios of the number of inpatient in medical treatment institution above county level,at county-level，and at township level were 26.7%,46.3%,and 27% in 2009；and were 30.4%,54.2%,and 15.4% in 2012. The constituent ratios of medical expenditure were 57%,34%,and 9% in 2009；and were 56.3%,37.9%,and 5.8% in 2012. The constituent ratios of hospitalization reimbursement were 49.2%,38.1%,and 12.7% in 2009；and were 46.3%,45.6%,and 8.1% in 2012.The average hospitalization reimbursement rates per time were 32.2%,41.5%,and 52.6% in 2009,and were 43.2%,63.1%,and 73.4% in 2012.Conclusion In the last decade,the rate of inpatient,medical expenditure and reimbursement to county level or higher level medical institutions increased significantly.However,the rate of township hospitals reduced year by year.The average hospitalization reimbursement rate per time in Jilin provinceincreased year by year,and the rate of township hospitals was the highest.

Objective To perform the prokaryotic expression,purification and identification of Cystatin of Cyprinus carpio var.Jian,and to further prepare a polyclonal antibody targeting Cystatin as the antigen. Methods The E.coli　BL21（DE3）,in which the pET-30a-Cystatin plasmid had been transferred,was induced by IPTG to express the recombinant protein,which was purified with Ni2+-NTA affinity chromatography.Then the purity of recombinant Cystatin and inhibitory activity was identified using high performance liquid and Azocasein respectively.The QuickAntibody-Mouse5W fast adjuvant and purified recombinant Cystatin were used to immunize Balb/C mice to get antiserum,whose titer was identified with sandwich ELISA and whose specificity  to recognize antigens was assessed with Western blotting and immunohistochemistry. Results The molecular weight of interest protein of prokaryotic expression was approximately 20 000.After Ni2+-NTA affinity chromatography,the recombinant Cystatin was shown as a single band on the SDS-PAGE and its purity was above 95%.Azocasein assay showed that 50% activityof papain was inhibited by the recombinant Cystatin of 0.014 μg.ELISA showed Cystatin antiserum with titer of 1∶512 000 was obtained.The results of Western blotting showed the Cystatin was absence in the total protein of E.coli　BL21(DE3) with transferred plasmid pET-30a,in contrast with transferred plasmid pET-30a-Cystatin and the sample in every process of purifying,the Cystatin was presence and showed a single band,and immunohistochemistry indicated the different tissues of Cyprinus carpio var.Jian showed different positive staining,the antiserum could specifically bind to the Cystatin expressed in both prokaryotic and eukaryotic systems. Conclusion The recombinant Cystatin of Cyprinus carpio var.Jian is expressed and purified successfully.And finally,the antiserum with high titer polyclonal antibody recognizing Cystatin of Cyprinus carpio var.Jian is obtained,which owns the high specificity and sensitivity.