Abstract:Objective To study  the effects of low dose radiation (LDR) on the expressions of intercellular adhesion molecule-1 (ICAM-1) mRNA and protein in kidney of diabetes mellitus (DM) mice and  illuminate that anti-inflammation of LDR is a main mechanism for diabetic therapy.Methods The healthy and right age C57BL/6J mice were divided into 4 groups including control,DM,LDR and DM/LDR.The mice in DM and DM/LDR groups were injected intraperitoneally with streptozocin (STZ) to set up DM models.The mice in DM/LDR and LDR groups were irradiated with 25 mGy every other day for 4 weeks.The expressions of ICAM-1 mRNA and protein in kidney were detected with RT-PCR and Western blotting 2,4,8,12 and 16 weeks after irradiation.Results The expressions of ICAM-1 mRNA and protein in kidney had no significant difference among 4 groups before LDR (P>0.05).The expressions of ICAM-1 mRNA and protein 2 weeks after irradiation with LDR were higher than those in the other 3 groups (P<0.05).The expressions of ICAM-1 mRNA and protein in the DM/LDR group 4 weeks after irradiation were also  significantly  higher than those in non-DM groups (P<0.05),but still
 significantly  lower than those in DM group (P<0.05),and the significant differences were kept to 16 weeks after irradiation.But the expressions of ICAM-1 mRNA and protein in LDR group were significantly higher than those in  control group(P<0.05).IHC assay showed that the glomerular and tubular in DM and DM/LDR groups were abnormal and the quantities of the positive staining cells were significantly increased compared with non-DM groups.However the damage of glomerular and tubular in DM/LDR was significantly supressed compared with DM gorup and the positive staining cells were also decreased.Conclusion  Under the circumstance of DM,LDR can significantly decrease the expressions of ICAM-1 mRNA and protein in mouse kidney to relief the inflammation reaction in kidney;but in normal condition,LDR can improve the immunity and promote the release of immune-related factors.It means that LDR can play different roles under different circumstance.

Abstract:Objective To establish the cell model expressing p53-175H and detect the gene silence effect of shRNA in H1299 (p53-/-) cell line. Methods PCR site-directed mutagenesis was used to gain p53-175H cDNA sequence,gene recombinant technique was used to construct the p53-175H expression vector and the vector was transferred into H1299（p53-/-）cells by liposome in vitro.The shRNA sequence targeting p53-175H was synthesized and inserted into retroviral vector Psuper to construct retro-Psuper-175HR.The retro-Psuper-175HR was transfected into packaging cell 293T using calcium phosphate co-precipitation and the viral soup containing sham-viral particle was used to infect H1299 cells directly.Western blotting was used to detect the protein expression of P53.Results The expression of P53 protein in the p53-175H cell model was positive.After transfection in H1299-175H cell model,the P53 protein expression was decreased.Conclusion The H1299-175H cell model is established successfully.Psuper-175HR gene silence vector can effectively suppress the expression of P53 mutant 175H in H1299-175H cells.

Abstract:Objective To study the expression of growth associated protein 43 (GAP-43) after optic nerve transection injury in visual system of neonatal rats and its significance. Methods The healthy neonatal SD rats (24 h) were randomly divided into normal control group and optic nerve transection injury group. Immunohistochemical staining technique was used to detect the expressions of GAP-43 in normal and optic nerve transection injury visual system of neonatal rats at 1,3,7,14,28,56 d after birth. Results The positive expression of GAP-43 in  normal control group was found in  visual system of  neonatal rats at developing stage and with  the  neurons mature the expression of GAP-43 was decreased(P<0.05). The positive expression of GAP-43 in  optic nerve transection injury group was found and was higher  than that in  normal control group.There was no significant change at 1 d,the expression of GAP-43 in optic nerve transection injury group was increased at 3 d and reached peak at 7-14 d,then it was gradually decreased to normal level until 56 d. Conclusion The expression of GAP-43 in visual system of neonatal rats changes with the development. The expression of GAP-43 is increased after optic nerve transection injury. This result suggests that GAP-43 may play an important role in the developing and regeneration of visual system.

Abstract:Objective To study the protective effects of gross saponins from tribulus terrestris (GSTT) on ischemia-reperfusion(I/R) injury in isolated rat heart and approach its mechanism of action.Methods Fifty-six Wistar rats were randomly divided into seven groups:control group,I/R group,ischemic preconditioning(IPC) group,adenosine group,and GSTT 200,100,50 mg?L-1 groups.The isolated hearts were subjected to 30 min ischemia and then followed by 40 min reperfusion.The contents of CK,LDH,AST，MDA and activity of  SOD in reperfused ischemic rat heart were detected.The histopathological changes of myocardium were observed by hematoxylin-eosin (HE) staining.Results Conpared with I/R group,the contents of CK,LDH,AST were decreased in GSTT 200 and 100 mg?L-1 groups（P<0.05  and P<0.01），and the contents of MDA in myocardium tissues were also decreased（P<0.05 and P<0.01）,and the activities of SOD were increased（P<0.05 and P<0.01）.GSTT could  alleviate the pathological lesion of myocardium tissues.Conclusion GSTT has protective effect on cadiocytes injured by ischemia-reperfusion,its mechanism may be concerned with resisting oxygen free radical.

Abstract:Objective To explore the way and technique to gain adequate seed cells by introducing human telomerase reverse transcriptase(hTERT) gene into rabbit chondrocytes.  Methods The chondrocytes of rabbit were isolated and cultivated,the expression vector pcDNA3.1/ zeo(+)-hTERT was transfected into chondrocytes, in control group it was zero load pcDNA3.1/ zeo(+),the positive clones were selected and amplified,the reverse transcription-polymerase chain reaction(RT-PCR) was used to detect the expression of hTERT mRNA, toluidine blue staining was used to analyze whether or not the chondrocytes transfected with hTERT gene possessed bionomics of chondrocytes.Flow cytometry examination was used to analyze cell cycle and karyotype.The condition of tumor formation of nude mouse was observed by tumorigenesis experiment.Results pcDNA3.1/ zeo(+)-hTERT was transfected into chondrocytes detected by RT-PCR on the RNA level.The morphology of chondrocytes infected with pcDNA3.1/ Zeo(+)-hTERT still kept polygon, which was same as the normal chondrocytes cultivated in vitro.After toluidine staining, polysaccharide was stained amethyst, small amethyst particles of proteoglycan were well-distributed.It suggested that the transformed cells still possessed the function of secreting cartilage correlated matrix.The multiplicate ability of transformed cells could be analyzed by measuring the cycle of cells.The multiplicate ability increased remarkably, the way of cell division was based on diploidy keeping the stabilization of karyotype.No tumorigenesis was found in nude mouse.Conclusion The proliferative ability of chondrocytes transfected with hTERT are enhanced obviously and the malignant transformation does not take place, they can serve as an excellent tool to study the biochemical and physiological aspect of chondrocytes and be used as seeding cells in tissue engineering.

Abstract:Objective To investigate the reversal effect of a novel polyamine conjugate,NMMB,on multidrug resistance (MDR) of myelogenous leukemia K562/ADM cells and  discuss the possible correlative mechanism.Methods K562 and K562/ADM cells in culture medium were treated with NMMB (10-1 000 mg/L) respectively.The inhibitory rates of these cells were measured by MTT assay.Non-cytotoxic dose of NMMB was determined. K562/ADM cells at logarithmic growth phase were randomly divided into ADM control,NMMB 2.5,5.0,10.0 mg/L groups.The inhibitory rate 50% (IC50) and the reversal index in all groups were determined.The effects of NMMB on ADM accumulation in K562/ADM cells and cell cycle  were examined by flow cytometry (FCM).Results The inhibitory rates were significantly increased when the cells were treated with different doses of NMMB (10-1 000 mg/L) in a dose-dependent manner.The available reversal concentration of NMMB was 12.5 mg/Land the reversal index was 4 folds on K562/ADM cells.ADM accumulation in K562/ADM cells was significantly increased (P<0.05).The cells were blocked at the period of G0/G1 (P<0.01).Conclusion NMMB has an effect on proliferation inhibition and MDR reversal of K562/ADM cell line.The reversal mechanisms of   NMMB may be due to increasing the accumulation of chemo-drugs in cells and arresting the cells at 0/G1 phase.

Abstract:Objective To investigate the electrophysiologic characteristics of volume - sensitive chloride currents in human trabecular meshwork cells and  clarify the relationship between this current and the volume-sensitive chloride channel.Methods Human trabecular meshwork cells were culivated in vitro,whole-cell patch-clamp was used to record the current of chloride channel of human trabecular meshwork cell membrane under the isotonic and hypotonic conditions,and the characteristics of the current and the effects of the chloride channel blockers NPPB and tamoxifen and ATP on the currents under  hypotonic condition were observed.Results Under isotonic condition,only weak and stable background currents were observed;Under  +100 and －100　mV voltage clamp,the outward and inward current density values（pA/pF） were 1.48±0.36 and －0.91±0.10 (n=6), respectively. The currents were increased rapidly by perfusing the cells with hypotonic solution and outward currents were prominent,the outward and inward current density values(pA/pF) were 19.94±0.87 and －0.53±0.41（n=6，P<0.01). When the solution was bathed by chloride channel blockers NPPB(100 mol/L),tamoxifen（50  mol/L）or ATP(2  μmol/L)for 20 min under hypotonic condition,the outward and inward current density values(pA/pF) were 7.61±0.50 and －3.70±0.25,8.21±0.63 and　－3.83±0.25,and 9.08±0.67 and 3.89±0.22, respectively;the outward and inward current density values were reduced significantly than those under  hypotonic condition（P<0.01）.Conclusion The outward chloride currents induced by extracellular  hypotonicity in human trabecular meshwork cells are prominent,which are sensitive to the change of cell volume,and can be inhibited by NPPB,tamoxifen,and ATP.The currents are in line with the characteristics of volume-sensitive chloride currents.

Abstract:Objective  To investigate the effects of phosphodiesterase inhibitors (PDEI) on atrial dynamics induced by  C-type natriuretic peptide (CNP) and the contents  of cyclic nucleotide (cAMP,cGMP) in isolated beating rabbit atria.Methods  After the rabbits had been anesthetized,the hearts were  removed rapidly.The left auricles were isolated and fixed on the atrial perfusion system.The atrial stroke volume and the pulse pressure were observed by CNP with or without PDEIs pretreatment.The contents of cAMP and cGMP were measured by radioimmunoassay.Results  ①Compared with control cycle group,CNP (30.0 nmol/L) obviously decreased the atrial stroke volume and pulse pressure (P<0.01) and increased the cGMP content(P<0.001),while the content of cAMP had no change (P>0.05). ②Compared with control cycle group,IBMX (1000.0 nmol/L),a non-selective inhibitor of PDE,significantly increased the atrial stroke volume,pulse pressure,cAMP and cGMP contents (P<0.001),but there were no differences  of the indexes mentioned above between  IBMX (1 000.0 nmol/L)　 plus CNP (30.0 nmol/L)group and IBMX group (P>0.05).③Compared with control cycle group,EHNA (30.0 nmol/L),an inhibitor of PDE2,obviously decreased the atrial stroke volume and pulse pressure (P<0.05) and cAMP content had no change (P>0.05).EHNA (30.0 nmol/L) plus CNP (30.0 nmol/L) showed similar roles with EHNA only.④   Compared with control cycle group,milrinone (1.0 nmol/L),an inhibitor of PDE3,significantly increased the content of cAMP (P<0.05) and slightly increased the atrial stroke volume and pulse pressure,but had no significant differences (P>0.05).CNP (30.0 nmol/L) obviously decreased the atrial stroke volume and pulse pressure (P<0.001),and increased the cAMP content (P<0.05) by pretreatment of milrinone. Compared with milrinone group,milrinone plus CNP decreased the atrial stroke volume and pulse pressure (P<0.001),and the cAMP content had no change(P>0.05).Conclusion CNP can inhibit atrial dynamics by increasing the content of cGMP,the different inhibitors of PDEs play different roles in the CNP-induced inhibition of atrial dynamics in isolated beating rabbit atria.

Abstract:Objective To study the effects of rosiglitazone on FOXO1 and TSC2 gene expressions,insulin secretory function,cell proliferation and apoptosis of pancreatic β cells under high concentration glucose condition.Methods The NIT-1 cells were put into plates (5×10 cells /well) and cultivated for 48 h,then they were randomly divided into treatment groups containing different concentrations of glucose as ollows:5.6,7.8,11.1,16.7,22.2,and 27.6 mmol/L groups. After cultivated for 24 h,they were intervented by 10^-5 mmol/L  rosiglitazone for next 24 and 48 h,then the supernatant was collected.The insulin level was evaluated by radio-immunity technique,the cell proliferation and apoptosis were detected by immunofluorescence staining and MTT assay respectivly.The expressions of FOXO1 and TSC2 mRNA were detected by semi-quantitative RT-PCR assay.Results ① Under different concentrations of glucose，after treated  with 10^-6-10^-5 mol/L rosiglitazone the proliferation of pancreatic β cells (NIT-1 cell line) was found(P<0.05)and the apoptotic rate of cells was increased in a dose-dependent manner （1×10^-5 mol/L rosiglitazone group＞1×10^-6 mol/L rosiglitazone group＞1×10^-7 mol/L rosiglitazone group）.② When under same dose of glucose,the insulin secretion level in 11.1 mmol/L group was much higher than those in other groups(P<0.05),but the insulin secretion level was reduced  gradually following the decrease of glucose concentration(11.1 mmol/Lgroup＞16.7 mmol/L group＞22.5 mmol/L group＞27.6　mmol/L group).The insulin secretion level in 5.6 mol/L group was the lowest.③ After intervention of 10^-5 mol/L rosiglitazone,the expression levels of both FOXO1 and TSC2 mRNA were significantly lower   than those in control group (5.6 mmol/L group＜11.1 mmol/L  group＜16.7 mmol/L  group＜22.5 mmol/L  group＜ 27.6 mmol/L group).When the glucose concentration was over 16.7 mol/L,the expressions of  FOXO1 and TSC2 mRNA were obviously higher than those in the groups with glucose concentration ≤11.1 mmol/L after the intervention of 10-5 mol/L rosiglitazone.Conclusion Rosiglitazone can improve the secretion function of pancreatic β cells and cell proliferation and alleviate insulin resistance by directly regulating FOXO1 and TSC2 expressions.

Abstract:Objective  To construct the shRNA lentiviral vectors targeting human NOB1 gene and detect its effect of gene silence in SKOV3 and HEY cells.  Methods The specific siRNA sequences targeting human NOB1 gene were cloned into pLVTHM lentiviral vector. After screening for the valid siRNA,the lentivirus particles were packaged and NOB1 specific shRNA was transmitted into SKOV3 and HEY cells. Then,real-time PCR was performed to determine the expression level of NOB1.  Results  PCR and sequencing results revealed that shRNA plasmids were correctly constructed. The virus with a titer of 8×10⁸ PU/L was successfully packaged.The  NOB1 expressions in SKOV3 and HEY cells were  down-regulated at mRNA level by virus infection. The expression levels of NOB1 mRNA was decreased by 73.5%  in SKOV3 and by 82.2% in HEY cells compared with negative control.

Abstract:Objective To study the expression of the  recombinant protein of keratinocyte growth factor -2 (KGF-2)  induced with  lactose,an alternative to IPTG induction agents,and  explore the possibility of the production of KGF-2 engineering. Methods The effects of the concentration,induction time and induction temperature of lactose  on the  expression  of the recombinant product were compared and  IPTG induction was used as a control to determine a more optimal conditions for lactose induction. Results For the production of recombinant KGF-2,lactose as an induction agent could also play a very good role;the A600 value  with lactose as induction agent was   higher than  IPTG, the expression level of induction product was equivanlent to IPTG induction. With 10　g/L lactose,at 33℃,induction for  6 h,the  induction effect was  better and the induction product  had good solubility and  could promote the cell growth. Conclusion Lactose can replace IPTG to express the  keratinocyte growth factor,and the  good effects are obtained.This study  provides a good experimental basis for the large production of KGF-2.

Abstract:Objective To construct the dual-subtype (H5/H7) influenza virus DNA vaccine and determine its immunogenicity.Methods Influenza virus DNA vaccine pVAX1-H5HA-H7HA co-expressing H5 subtype hemagglutinin (HA) and H7 subtype HA was constructed.The expressed target proteins in BHK cells were identified by IFA.Immunized mice were divided into four groups(15 mice in each group):pVAX1-H5-H7,pVAX1-H5,pVAX1-H7 and pVAX1 control groups,and the serum antibodies were detected by ELISA,the number of splenic T lymphocyte subgroups was determined by flow cytometry.Results The product of constructed dual-subtype (H5/H7) influenza virus DNA vaccine in BHK cells stimulated the expression of green immunofluorescence of fluorescent antibody,and the pVAX1 empty vector transfected BHK cells had no immunofluorescence.The immunized mice produced the serum specific antibodies.The number of splenic T lymphocyte subgroups CD4+ and CD8+ was  higher than that in control group（P<0.05）.Conclusion Dual-subtype (H5/H7) influenza virus DNA vaccine is successfully constructed and this DNA vaccine can induce specific cellular and humoral immune responses in mice.

Abstract:Objective To construct and screen shRNA plasmids of HSPA8 with high interfering ability and  provide the material for further  researching on the effect of HSPA8 gene on breast cancer.Methods In accordance with the sequence of HSPA8 gene in GenBank,four interfering gene fragments and four shRNA interfering plasmids（pGPU6/GFP/Neo-349,pGPU6/GFP/Neo-818,pGPU6/GFP/Neo-931,pGPU6/GFP/Neo-1180）were constructed.The plasmids were transfected  into  human breast cancer cell MCF-7 respectively,and negative control group and blank control group were set up.The HSPA8 positive expressing cells were screened by G418.The interfering efficiencies of the plasmids on HSPA8 gene  were detected  at the mRNA and  protein levels by RT-PCR and Western blotting.Results The RT-PCR results   showed  that at the mRNA  level  in four transfected groups the expression levels were decreased by 25.0%,37.5%,43.7%,and 68.5%,respectively,compared with untransfected group.The Western blotting results showed that the differences of the protein expression levels between transfected groups and blank control group were significant(P<0.05).The interfering efficiency of  pGPU6/GFP/Neo-1180 plasmid was the highest.Conclusion The high ability shRNA interfering plasmids of HSPA8 gene are constructed successfully.The pGPU6/GFP/Neo-1180 interfering plasmid may provide a basis  for  further study on the occurrence and  development of breast cancer.

Abstract:Objective To study the mechanism of the changes of proliferation rate induced by the proteasome inhibitor Lactacystin(LAC)   through  observing the effect of    LAC on the proliferation rate of glioma C6 cells, and provide a theoretical basis for clinical treatment of glioma. Methods The cultured rat glioma C6 cells at the logarithmic growth phase were divided into control group,LAC 2.5 μmol/L group,LAC 5.0 μmol/L group and LAC 10.0 μmol/L Group.MTT assay was used to detect the proliferation rate of C6 cells,FCM was used to detect the death and mitochondrial membrane potential of C6 cells and RT-PCR was performed to examine the Bax and bcl-2 mRNA expressions,Western blotting was used to determine the Bax,bcl-2 and P65 protein expressions.Results Compared with  control group,the proliferation rates of C6 cells,mitochondrial membrane potential and  the level of P65 protein expression in LAC 5.0  and LAC 10.0 μmol/L groups were  reduced (P<0.05),while the apoptotic rate,Bax/bcl-2 mRNA and protein ratio were increased (P<0.05).Conclusion  LAC can inhibit the proliferation rate of C6 cells by inclucing the apoptosis of C6 cells,which may be related to the mitochondrial pathway,in addition,NF-κB signaling pathway may also affect the proliferation rate of the C6 cells in this process.

Abstract:Objective To examine the nerve connections of the sexually dimorphic nucleus of the preoptic area (SDN-POA) in the rat, and explore the function of  SDN-POA.Methods A total of 30 transgenic rats were used in the study，which expressed enhanced green fluorescent protein (EGFP) under the control of an estrogen receptor α gene promoter 0/B.The male and female rats (n=15, respectively) were randomly divided into three groups, and Fluoro-Ruby, a fluorescent tracer, was stereotaxically injected into the ventromedial hypothalamus (VMH), bed nucleus of the stria terminalis (BST) and posterodorsal nucleus of the amygdale (MePD) by iontophoresis, respectively.Five days later, the rats were perfused, and the sections containing SDN-POA region and site of injection were cut on a freezing microtome.the sections were viewed to assess extent of tracer deposition in the injection site and to reveal the EGFP or Fluoro-Ruby fluorescence in SDN-POA under fluorescence microscope.Results In male and female transgenic rats,  the cells labeled with EGFP were specifically distributed in the SDN-POA, and the volume and the number of cells in SDN were larger in the male than female and it demonstrated the sex difference.In the groups injected with tracer into the VMH and BST, the cells labeled with Fluoro-Ruby were determined in SDN.The labeled cells from VMH were distributed in the later of the SDN and the labeled cells from BST were dispersedly distributed in SDN in both sexes.In the group injected with tracer into the MePD, the labeled nerve fibers，not neurons，were discovered in SDN in both sexes.Conclusion SDN-POA is an important center nucleus to determine sexual dimorphism, and it would receive the afferent nerve fiber from MePD and send the efferent nerve fiber to VMN and BST to regulate reproductio
n.

Abstract:Objective To construct and express a gene engineer bacteria which expresses human  VEGF121 and provide a experimental foundation for targeting therapy of tumor vessel.Methods SUMO-CM-VEGF121 fusion genes were obtained by PCR.The VEGF121 was fused with  SUMO-CM by PCR,and the fused gene was transformed into E.coli Rosetta-gami(DE3).Soluble proteins were obtained at high level by IPTG.The fused protein was purified by DEAE sepharose and Ni-NTA affinity chromatography.Once cleaved from SUMO,the purity of CM-VEGF121 was obtained.Results The acquired gene fragments of SUMO-CM-VEGF121 were identified by digestion and DNA sequencing,and the fusion gene was 774 bp;the best condition for soluble protein was to cultivate the bacteria of  Rosetta-gami (DE3)/pET22b-SUMO-CM-VEGF121 at 20℃ for 24 h,with a soluble expression level at 21%.Western blotting result showed that this protein had the same immunogenicity with human VEGF antibody.Conclusion The prokaryotic expression vector pET22b and host bacterium Roestta-gami are most suitable for CM-VEGF121,which solves the problem of low expression and insolubility.

Abstract:Objective To investigate the influence of purine nucleotide on adenosine deaminase (ADA) activity and gene expression in testis and epididymis tissues of heroin dependence and withdrawal rats.Methods The rat models of heroin dependence and withdrawal were set up.80 male Wistar rats were randomly divided into 8 groups (10 rats of each group):control group(C),nucleotide group(N),heroin administration group(H),heroin and nucleotide group(HN),3 d withdrawal group(W3),9 d withdrawal group(W9),nucleotide 3 d group(N3),nucleotide 9 d group(N9).The adenosine deaminase activity and gene expression in testis and epididymis tissues of rats in various groups were detected.Results Compared with control group,the ADA activities in heroin administration group and 3 d withdrawal group in testis and epididymis tissues were increased significantly (P<0.05);there was no obvious  difference of ADA activity between other groups and control group（P>0.05）.The ADA mRNA level in testis tissue in heroin group was significantly higher than that in control group (P<0.05);the ADA mRNA levels in epididymis tissue in heroin group,3 d withdrawal group and 9 d withdrawal group were significantly higher than that in control group (P<0.05);and there was no obvious difference of ADA mRNA level between other groups and control group（P>0.05）.Conclusion Heroin can enhance constantly the adenosine deaminase activity and gene expression in testis and epididymis tissues of rats.Purine nucleotide compensation can oppose the effect of heroin.

Abstract:Objective To study the changes of hemodynamics and electrocardiogram of the acute myocardial infarction models  with blood stasis and the simple acute myocardial infarction models in rats  and elucidate the difference between the acute myocardial infarction models  with blood stasis and the simple acute myocardial infarction models in rats.
Methods Wistar rats were randomly divided into control  group,acute blood stasis models group,acute myocardial infarction and sham operation group,acute myocardial infarction models group and  acute myocardial infarction models with blood stasis group(n=12).The acute myocardial infarction models under the status of the acute blood stasis in rats were built to determine the parameters of the left ventricular systolic pressure(LVSP),left ventricular end-diastolic pressure (LVEDP),maximum left ventricular rise rate and maximum left ventricular fall rate(±dp/dt max),and the pressure of carotid artery including systolic blood pressure (SBP),diastolic blood pressure (DBP) and mean artery pressure (MAP).In addition,the parameters of electrocardiogram (ECG) including heart rate (HR) and ST segment with V5 le
ad and Ⅱlead were also measured.Results  ① When the acute myocardial infarction models with blood stasis in rats and the simple acute myocardial infarction models were comparatively studied,the results showed that there were no obvious differences between them in SBP,DBP,MAP,LVSP,LVEDP and ±dp/dt max(P>0.05);② The increasing extents of the HR and ST segment with V5 lead and Ⅱlead of the acute myocardial infarction models with blood stasis in rats were much higher than those of the simple acute myocardial infarction models(P<0.05 or P<0.01).Conclusion The obvious differences are shown on the parameters of ECG,but there are  no obvious differences in the hemodynamics when the acute myocardial infarction models with blood stasis and the simple acute myocardial infarction models are comparatively studied.The results  show that it is much reasonable to evaluate pharmacodynamics of traditional Chinese drug which treat thoracic obstruction cardialgia in the acute myocardial infarction models  with blood stasis in rats.

Abstract:Objective To investigate the influence of the recombinant plasmid Psilencer1.0-U6-siRNA-stat3  on neointima formation of vein graft and develop a new way to prevent restenosis of the vessel graft after arterial bypass. Methods 40 Wistar rats were divided into 2 groups(n=20):  experiment group and control group.The recombinant plasmid Psilencer1.0-U6-siRNA-stat3  (exeperiment group) and scramble plasmid (control group) were combined with the liposomes Lipofectimine 2000 and were transfected into vein graft mediated by bioprotain gel via adventitia.On day 3，7，14，28，the vein graft interposed into the arterilized environment were extracted after irrigating 4% formaldehyde through left ventricle in 100 mmHg.The change of neointimal thickening was analyzed by pathological(HE) study and the proliferation of vascular smooth muscle cells(VSMCs) in neointima was detected by immunohistochemistry (proliferating cell nuclear antigen,PCNA). Results The immunohistochemistry results showed that,on day 7 and 14 after operation,there were more PCNA positive cells in neointima of vein graft in control group than that in  experiment group.The PCNA index in experiment group（23.3±2.8） was lower than that in control group（31.3±4.7）( P<0.05) 7 d after operation. The PCNA index  in experiment group(15.6±0.4) was also lower than that in control group (27.2±5.7)( P<0.01) 14 d after operation.The morphological result showed that the thickness of neointima experiment group(0.50μm±0.02μm) was thinner than that in control group(0.90μm±0.02μm) ( P<0.05). Conclusion The recombinant plasmid Psilencer1.0-U6-siRNA-stat3  can inhibit  the neointimal formation of vein graft obviously.

Abstract:Objective To investigate the potentiality of bacterial cellulose(BC) as the scaffold of tissue engineering corneal stroma . Methods Rabbit and human corneal stromal cells were separated and seeded into a BC membrane. The compound membrane with rabbit stroma cells was allogeneil grafted and examined by corneal confocal microscopy,anterior OCT and histological method and immunohistochemical staining 1,4 and 8 weeks after operation. Results The rabbit and human corneal stromal cells grew well in the BC membrane. One week after grafting,the inflammatory reaction and new vessels were not observed in the cornea. After four weeks,the new vessels could be found around the membrane without edema and ulcer on the surface. The rest cornea was still transparent. The stromal cells proliferated actively around the membrane observed by corneal confocal microscopy 8 weeks after transplantation. The quantity and appearance of proximalis corneal endothelium were normal. Anterior OCT displayed that BC compound membrane degraded gradually .The boundary between membrane and normal corneal tissue was vague 4 weeks after operation and similar to normal cornea in density 8 weeks after operation. BC compound membrane approached to normal corneal stromal tissue by ex vivo histology.The inflammatory cell infiltration and new vessels could be detected without necrosis or dissolution. Conclusion  BC has a good biocompatibility and degradation without cytotoxicity . It can be used as a biological scaffold for tissue engineering corneal stroma.

Abstract:Objective To study the anatomy of adult anterior interosseous nerve and its muscular branches and provide the morphological basis for the surgical treatment of the anterior interosseous nerve syndrome and repair of the median nerve and ulnar nerve.Methods By the method of the anatomical dissection,the location of the anterior interosseous nerve and its muscular branches in the forearm from 30 upper extremities of adult cadavers were observed with visual inspection especially while its basic anatomical data such as length and diameter were measured.Results The anterior interosseous nerve arose from the median nerve (45.7±10.5)mm and gave off the muscular branches (79.06±7.49)mm below the line between the medial and lateral epicondyles of humerus.It was located in 13.2%－32.8% of the forearm. The number of the branches to the flexor pollicis longus (FPL,93.3%) and pronator quadrates (PQ,100%) was 1,the number of the branches to the flexor digitorum profundus (FDP,86.7%) was 4.The muscular branches of the anterior interosseous nerve were mainly located in 27.6%－88.1% of the forearm.The branches to FPL was located in 27.6%－48.3%;the branches to FDP was located in 32.8%－49.9%;the branches to PQ was located in 41.3%－88.1%.Conclusion  This study may provide the anatomical basis for the surgical treatment of the anterior interosseous nerve syndrome and
 repair of the median nerve and ulnar nerve.The relative data and location method provided in this study  are helpful to improve the accuracy and security of operation.

Abstract:Objective To investigate the relationship between multi-drug resistance relative protein-2 (MRP-2) and leukemia multi-drug resistance，and  study its effect on leukemia multi-drug resistance.Methods The expressions of MRP-2 in leukemia cell line K562 and leukemia multi-drug resistance cell line K562/A02 were detected by PT-PCR.Anti- and oligonucleotide of MRP-2 were transfected by lipidosome into K562/A02,intra-cellular doxorubicin (ADM) fluorescence intensity,resistance index,and the expression of MRP-2 mRNA were detected by MTT,flow cytometry,and RT-PCR.Results The expression of MRP-2 in K562/A02 was higher than that of K562(P<0.05).After transfected with anti-oligonucleotide MRP-2,the ADM fluorescence intensity of K562/A02 was increased,and the resistance index was significantly decreased,and the expression of MRP-2 mRNA was   decreased by 67.5% (P<0.05) compared with untransfected cells.Conclusion The high-level expression of MRP-2 results in the reducing of chemical drug concentration,which may be one of the  mechanisms of leukemia multi-drug resistance.

Abstract:Objective To detect the expressions of triggering receptor expressed on myeloid cells-1(TREM-1),TNF-α and IL-8 of foam cells derived from macrophages.Methods Human U937 cell strain was induced by PMA and then was divided into PMA  group,LDL  group and ox-LDL  group.RT-PCR was applied to measure the mRNA expression of TREM-1 in each group and ELISA was used to detect the contents of TNF-α and IL-8  in culture supernatant in each group.Results Compared with PMA group and LDL group,the TREM-1 mRNA expression in ox-LDL group was significantly increased(P<0.05);the TNF-α and IL-8 contents in culture supernatant in ox-LDL group were  also significantly increased(P<0.05).Conclusion TNF-α,IL-8,and TREM-1 maybe play an important synergetic effect in the process of atherosclerosis.

Abstract:Objective To investigate the kinds and contents of trace elements in marten’ s heart and chicken’ s heart (which is easily confused with marten’ s heart),and provide a scientific method to identify them.Methods The various trace elements in marten’ s heart and chicken’s heart were  measured by inductively coupled plasma atomic emission spectroscopy（ICP-AES）after they were treated with concentrated nitric acid.Results  Eight kinds of trace elements were found in marten’s heart.The magnesium content was 685.31 mg/kg and it was the highest among the detected trace elements.Nine kinds of elements were found in chicken’s heart,but the potassium content was 11 829.7 mg/kg and the highest in it.The detection line of different elements was 0.4-12.1  μg/L,the recovery rate of the method was 92.0%-111.9%,the RSD(n=5) was 0.5%-3.8%,and all R values of the different standard curves were more than 0.999 9.Conclusion There are great differences in kinds,contents and the content ratios of different elements in marten’ s heart and chicken’ s heart. The results suggest that ICP-AES can be used for identifying marten’ s heart and chicken’ s heart.

Abstract:Objective To investigate the effects of different concentrations of lithium(LiCl) on neonatal rat cardiomyocytes exposed to high glucose,and study the  protective effects of canonical Wnt signaling pathway.Methods The ventricular myocardium of  neonatal rats about 1-3 d old was obtained.Then the primary culture cardiomyocytes were randomly divided into 4 groups:control group,high glucose damaged group,LiCl  5 and 10 mmol/L group.The morphological of cardiomyocytes were observed by inverted microscope,the apoptotic cardiomyocytes were tested by terminal deoxynucleotidyl transferase mediated dUTP_biotin nick end labeling(TUNEL),immunocytochemistry staining was used to detect the protein experssions p-GSK-3β and β-catenin.Results  Compared with high glucose damaged group, the pulsation frequency of cardiomyocytes tended towards nomal,the apoptotic cardiomyocytes were decreased（P<0.05）,the expressions of p-GSK-3β and β-catenin were increased（P<0.05）with the increasing of the concentration of LiCl in LiCl 5 and 10 mmol/L groups.Conclusion  LiCl can significantly inhibit the injury of cardiomyocytes induced by high glucose,its mechanism may be related to the activation of canonical Wnt signaling pathway．

Abstract:Objective To investigate the synthesis and secretion of collagen type Ⅰ(Col Ⅰ) in human adipose-derived stem cells(hADSCs) during osteogenic differentiation and induction,and approach the efficacy of hADSCs as seed cells for bone regeneration engineering.Methods hADSCs were separated,cultivated and amplified in vitro and were identified by cell phenotype detection. Multi-lineage differentiation potential was assessed by histological staining such as oil red O for lipid accumulation and alizarin red S for mineralization nodules during adipogenic and osteogenic induction. Osteogenic differentiation and induction group was divided into five subgroups:0,7,14,21 and 28 d groups. Alizarin red S staining,Van Gieson collagen fiber staining and Col Ⅰ immunofluorescence were used to detect the expression of Col Ⅰ. FV Viewer 1.7 software was used to analyze the data. Results The positive stainning of oil red O was found 14 d after adipogenic induction. For the osteogenic induction groups,the calcified nodules and the collagen fiber red staining were found in 21 d group. The expression of Col Ⅰ was found in 21 d group for all cells and  strongly positive expression in 28 d group. Compared with 0 d group,the expression of Col Ⅰ in osteogenic induction group was significantly increased from 7 d to 28 d（P<0.05). Conclusion During osteogenic induction of hADSCs,all cells have osteogenic capability and the hADSCs have outstanding capability of osteoinductive and osteogenesis. hADSCs may be useful in  clinical cell-based therapy for bone regeneration engineering.

Abstract:Objective To construct μ2 subunit of adaptor protein 2 (AP2-μ2) small interfering RNA( siRNA) and expression vectors, and detect their silencing effects. Methods AP2-μ2 mRNA targeted hairpin siRNAs were devised and three oligonucleotide strands of DNA fragments encoding the above siRNAs were synthesized. After annealing of the complementary strands, the DNA fragments were cloned into pSilencer 3.1 neo H1 plasmid, followed by amplification in E. coli and DNA sequencing. The transfected cells were divided into five groups: control, scramble plasmid, AP2 plasmid-1, AP2 plasmid-2 and AP2 plasmid-3. Three groups of recombinant plasmids and the negative control siRNA plasmid(scramble plasmid)  were transfected into rat cardiac cell line H9C2 by liposome transfection method, and the expression of AP2-μ2 mRNA was examined by RT-PCR. Results The 　synthesis of six single-stranded oligonucleotide showed multiple bands by gel electrophoresis. After annealing respectively, the product by gel electrophoresis showed a single band.  After double-stranded DNA templates were connected with plasmid, they digested by restriction enzyme of BamHⅠ and Hind Ⅲ, the 4300 bp and 66 bp fragments were seen by gel electrophoresis. DNA sequencing analysis showed that the connecting sequence of nucleic acid sequences was same as the designed, indicating three  AP2-μ2 siRNA vectors were successfully constructed. RT-PCR results showed that in  AP2 plasmid-1 group, the AP2-μ2 gene expression level in transfected H9C2 cells reduced by 96% compared with  negative control group. Conclusion Three kinds of AP2-μ2 siRNA vectors are successfully constructed, and AP2 plasmid-1 transfected cells can significantly inhibit the AP2-μ2 gene expression.

Abstract:Objective To investigate the effect of  oligodeoxynucleotide (ODN) on  differentiation of  rat bone marrow mesenchymal stem cells(BMSCs) to osteoblasts.Methods BMSCs at the  third generation  were incubated for 24 h before 12 different ODN were added.The levels of alkaline phosphatase(ALP),reflecting the degree of osteogenic differentiation were detected in four replicate wells of BMSCs stimulated with each ODN with different concentrations( 4.0,2.0,1.0 and 0.5 mg/L) at different time points(24,72 and 120 h).Results There were no significant differernces of the mean level of ALP between 4.0 and 0.5 mg/L ODN groups and PBS control group;but in 2.0 and 1.0 mg/L ODN groups,there were two ODN which could increase the ALP levels at different time points(24,72 and 120 h)(P<0.01). Conclusion Two ODN of 12 different ODN can promote the diffentiation of BMSCs to osteoblasts.It suggests that ODN with special sequence characteristic can significantly promote the differentiation of rat BMSCs to osteoblasts.

Abstract:Objective To study the effect of astragaloside Ⅳ (AsⅣ) on myocardial ischemia-reperfuion injury in rats and clarify its mechanism.Methods The models of myocardial ischemia-reperfusion injury in rats were established.30 male Wistar rats were divided into sham-operation group,ischemia-reperfusion model group,positive drug NIC group，AsⅣ group(5 mg/kg,10 min before ischemia),AsⅣ+chelerythrine chloride(CHE) group （3 mg/kg）.The activities of serum LDH and SOD and contents of serum MDA and NO in rats were measured.The morphological changes of cardiocytes in rats were observed by light microscopy.The protein expression of PKCε was analyzed by Western blotting.Results The LDH activity and MDA content in AsⅣ group were lower than those in model  group (P＜0.05 or P＜0.01),and the SOD activity and NO content were higher in AsⅣ group than those in model group (P＜0.01).HE staining results confirmed that AsⅣ could obviously ameliorate cell structure and reduce inflammatory cell infiltration.AsⅣ could increase the protein expression of PKCε in membrane.CHE abolished the protective effect of AsⅣ partly, compared with AsⅣ group,the LDH activity and MDA content were increased(P＜0.05),the SOD activity was decreased(P＜0.05),the protein expression of PKCε in membrane was decreased(P＜0.05),the cardiocytes had serious edema and the cytoplasm was stained slightly in AsⅣ+CHE group.Conclusion AsⅣ possesses an obviously protective effect on myocardial ischemia-reperfusion injury in rats by increasing NO content. PKCε may be the down stream of NO which is involved in the transduction of AsⅣ.

Abstract:Objective To study the effect of 20(s)-proto-panaxdiol（PPD）on apoptosis of Siha cells in vitro,and clarify the mechanism of its anticancer action. Methods Siha cells cultivated in vitro were treated with alcohol,40 μg/L Platinol(Pt),10 μmol/L PPD,20 μmol/L PPD,40 μmol/L PPD for 48 h,respectively.The morphological changes of Siha cells were observed under optical microscope.Apoptosis was detected by Annexin V-EGFP /PI staining and TUNUL staining.The transcription and the protein expression of bax were analyzed by RT-PCR and Western blotting.Results After treated with PPD,the cells were round,recovery,and broken.Green and red fluorescence was shown in Annexin V-EGFP/PI staining,green fluorescence was evident in TUNUL staining.The transcription and the protein expression of bax were up-regulated.Conclusion PPD may induce the apoptosis of Siha cells,and its mechanism may be related to the up-regulation of bax expression.

Abstract:Objective To observe the effects of  20(s)-protopanaxadiol (PPD) on the growth of prostate cancer cells RM-1 and explore the mechanism of its action.Methods The mouse models with prostate cancer were constructed with RM-1 cells.36 mice C57 were randomly divided into three groups:model group(fed with  an equal volume of olive oil once a day,an equal volume of normal saline injection  intraperitoneal  once a week),paclitaxel(Taxol) group(20 mg/kg,intraperitoneal injection  once a week),PPD group(20 mg/kg,ig once a day),continuous administration of 4 weeks.The tumor size was measured once every 3 d,and the morphological changes of tumor were observed,the animals were killed over the fourth weekend.Observation:the general state of animal tumor weight,tumor volume,inhibitory rate,mortality;and the expression of Cyclin D1 gene was detected with RT-PCR and Western blotting.Results Compared with model group,the tumor growth of mice in other groups significantly slowed down,and the mice in PPD group had bright hair.The mice in  model group  scrambled and moved slowly with tumor ulceration,erosion and hemorrhage.The tumor volume of the mice in PPD group was significantly smaller than that in model group(P<0.05) 13 d after administration,and it was also significantly smaller than that in Taxol group(P<0.05) 21 d after administration.Compared with model group,the tumor weight of the mice in PPD group was significantly decreased(P<0.05),but the inhibitory rate was significantly increased(P<0.05),no death.Compared with Taxol group,the inhibitory rate in PPD group was significantly increased(P<0.05),the mortality was significantly decreased(P<0.05).The RT-PCR and Western blotting results showed that Cyclin D1 gene and protein expressions in treatment  groups were significantly reduced(P<0.05)compared with model group;compared with Taxol group,the expression of Cyclin D1 in PPD group was significantly decreased(P<0.05),but there was no significant difference of Cyclin D1 protein expression between PPD group and Taxol group(P>0.05).Conclusion PPD can significantly inhibit the growth of prostate cancer cells RM-1 and its mechanism may be related to the down-regulation of the expression of Cyclin D1.

Abstract:Objective To detect the expressions of matrix metalloproteinase-26 (MMP-26),metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase-4 (TIMP-4) in the cervical carcinoma tissues and analyze their correlation with clinical features.Methods The expressions of MMP-26,MMP-9 and TIMP-4 were detected in 10 cases of normal cervical tissues,6 cases of cervical carcinoma in situ and 79 cases of invasive carcinoma of cervix by immunohistochemistry.Results MMP-26 showed strongly positive staining in cervical carcinoma in situ and invasive carcinoma of cervix,their positive rates were 83.33% and 82.47%, respectively;there were significant differences compared with normal cervix (P<0.05).The expression of MMP-26 was closely correlated with the depth of myometrial invasion and lymph node metastasis(P<0.05).The positive rate of MMP-9 in cervival carcinoma tissue was 68.35%.The expression of MMP-9 was strongly correlated with pathological grading,its expression in grade Ⅲ was significantly higher than those in grade Ⅰ and Ⅱ（P<0.05）.The expression of TIMP-4 gradually decreased from normal cervical tissue to carcinoma in situ and invasive carcinoma,but no significant difference（P>0.05）.The expression of TIMP-4 was strongly correlated with clinical stage,its expression in early stage was significantly higher than that in late stage（P<0.05）.The expression of MMP-26 showed negative correlation with TIMP-4（r_s=－0.259,P<0.05）,while there was no correlation between MMP-26 and MMP-9（r_s=0.140,P>0.05）.Conclusion The expressions of MMP-26,MMP-9 are increased in cervix carcinoma tissue.They promote the local invasion and metastasis of cervix carcinoma tissue .The expression of TIMP-4 is decreased in cervix carcinoma tissue.Their application together could be a marker to monitor the prognosis of vervical carcinoma tissue.

Abstract:Objective  To elucidate the genetic association between TNXB gene polymorphisms and schizophrenia in a Han descent population in the north of China.Methods Genomic DNA was isolated from the whole blood samples.A single nucleotide polymorphism,rs204887（Rsa I site） present in the TNXB gene,was detected using PCR-based restriction fragment length polymorphism analysis among 779 Chinese Han patients with schizophrenia and 983 ethnicity-matched healthy controls.The Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness-of-fit χ test.The frequencies of alleles and genotypes of TNXB gene and the relationship between allelic frequencies and the clinical phenotypes of schizophrenia were statistically computed.Results The goodness-of-fit χ2 test result showed that the genotypic distributions of rs204887 did not deviate from Hardy-Weinberg equilibrium in both patient group and control group（P>0.05）.The χ test result did not show allelic association and genotypic association for rs204887（P>0.05）.There were no correlations between allelic or genotypic frequencies and the clinical phenotypes of schizophrenia（P>0.05）.Conclusion TNXB locus may not be associated with schizophrenia in a Han descent population in the north of China.

Abstract:Objective To study the expressions of TGF-β and P53 protein in prostatic carcinoma tissue and their clinical significances.Methods Immunohistochemical method was used to detect the expressions of TGF-β and P53 protein in 36 cases of prostatic carcinoma.Results 36 cases of prostatic carcinoma included 10 cases of well differentiation,16 cases of moderate differentiation,10 cases of poor differentiation.The positive expression rates of TGF-β in well,moderately,poorly differented prostatic carcinoma tissues were 30%(3/10),37.5%(6/16),and 70%(7/10);the positive expression rates of P53 protein in well,moderately,poorly differented prostatic carcinoma tissues were 20%(2/10),25%(4/16),and 50%(5/10). The expression rates of TGF-β and P53 protein in well and moderately differented prostatic carcinoma tissues were lower than those in poorly differented prostatic carcinoma tissues(P<0.05).Conclusion The expressions of TGF-β and P53 have a negative correlation with the differented degree of prostatic carcinoma and the expressions of TGF-β and P53 may help to judge the pathologic grade and predict the prognosis of prostatic carcinoma.

Abstract:Objective Through detecting the expression of dentin matrix protein 1 (C-DMP1) in pleomorphic adenoma and mucoepidermoid carcinoma tissues to investigate the role of expression of C-DMP1 in the oncogenesis of salivary gland tumor.Methods The immunohistochemical method was used to observe the expression of C-DMP1 in 30 cases of benign salivary gland tumor and 30 cases of malignant salivary gland tumors.Results In mucoepidermoid carcinomas,the expression of C-DMP1 in the epidermoid cells was strongly positive,in the nuclei part of the intermediate cells was weakly positive,while the mucous cells were negative,the strongly positive expression rate was 73.3%;In pleomorphic adenoma,part of the nuclei of ductal cells,outer myoepithelial cells and cartilage-like cells were weakly positive,with only a few strongly positive,the strongly positive expression rate was 6.6%.There was significant difference in the stongly positive expression rate of C-DMP1 between benign and malignant tumors in salivary gland (P<0.05).Conclusion The expression of C-DMP1 in the benign and malignant salivary gland tumor tissue  has different changes in its distribution,it suggests that C-DMP1 may participate in the oncogenesis of salivary gland tumor.

Abstract:Objective To investigate the corrrelation between hepatitis B virus X protein(HBx) and vascular endothelial growth factor (VEGF) expressions and angiogenesis and metastasis in HBV related hepatocellular carcinoma tissue.Methods Immunohistochemistry was used to detect the expressions of HBx,VEGF and CD34 in 84 cases of HBV related hepatocellular  carcinoma and 22 case of non-HBV related hepatocellular carcinoma tissues,the microvascular density(MVD) was recorded under optical microscope.Results The positive expression rate of HBx in 84 cases of HBV related hepatocellular carcinoma tissues was 73.81% (62/84),the negative rate was 26.19% (22／84).The positive expression rate of VEGF   in 62 cases of HBx positive expression was 74.19 % (46／62),which was significantly higher than those in   22  hepatocellular carcinoma tissues with HBx negative expression (36.36%,8／22) and  22 non-HBV related hepatocellular carcinoma tissues(36.36%，8/22),while there was   no statistically significant difference of VEGF prositive expression between HBV related hepatocellular carcinoma tissue with   HBx negative expression and non-HBV related hepaocellular  carcinoma tissue(P>0.05).HBx has positive correlation with VEGF（Rs= 0.552,P<0.05).The expressions of HBx and VEGF in well differentiation group were higher than those in  poor differentiation group(P<0.05),and the expressions of HBx and VEGF in metastasis group were higher than those in non-metastasis group.The MVD in HBx positive expression group was significantly higher than those in HBx negative expression group and non-HBV related hepacellular carcinoma  group (P<0.01),the MVD in metastasis group was higher than that in non-metastasis group (P<0.01),and the MVD in portal vein invasion group was higher than that in non-invasion group (P<0.05).Conclusion HBx and VEGF widely express in HBV related hepatocellular carcinoma tissues,and they have positive relationship.HBx may play an important role in angiogenesis and metastasis in hepatocellular carcinoma by up-regulating the VEGF expression.

Abstract:Objective To investigate the effects of Kudiezi injection on stent thrombosis and the levels of matrix metalloproteinase (MMPs) and thromboxane B2 (TXB2)in elderly patients with percutaneous coronary intervention (PCI), and  investigate the mechanism of Kudiezi on the decrease of stent thrombosis.Methods  Forty elderly patients were divided into two groups(Kudiezi group and control group) after PCI.Kudiezi were administered into patients in Kudiezi group and the patients in control group were treated with  regular medication.The angioraphic and clinic follow-up outcomes of 40 elderly patients with PCI there retrospectively analyze . Stent thrombosis (ST)was confirmed by angiography.The levels MMPs and TXB2 in Kudiezi group (n=20) and control group(n=20) were determined before stent implantation and after 6 months. Major cardiac events （restenosis,cardiac death,myocardiac infarction,revasculation）were observed during follow-up. Results The levels of MMPs and TXB2 in Kudiezi and control group decreased significantly after PCI.The levels of MMPs and TXB2 in Kudiezi group were less than that in control group after PCI.The levels  of MMPs and TXB2 in all patients group were signifantly different between pre-procedure and post-procedure (P<0.05). Within 6 months of stent implantation,the occurrence of ST was less in Kudiezi group than that in control group (P<0.05).Conclusion Kudiezi injection can deminish  the onset of stent thrombosis  and decrease the levels of  MMPs and TXB2.

Abstract:Objective To investigate the effects of Naoshukang(NSK) capsule on the hemorheology and thrombosis in rats and elucidate its protective mechanism on brain ischemia. Methods 40 Wistar rats were randomly divided into 5 groups:control, NSK 2 g/kg, NSK 8 g/kg, NSK 16 g/kg and Buchang Naoxintong(BCNXT) 2 g/kg groups.Thrombosis time of carotid artery was observed under electrical stimulation.Another 40 Wistar rats were randomly divided into blood stasis model, NSK 2 g/kg, NSK 8 g/kg, NSK 16 g/kg and BCNXT 2 g/kg groups.The thrombus length in vitro, dry weight and wet weight, ADP-induced platelet aggregation rate, clotting time, whole blood and plasma viscosities were measured. Results Compared with control group, thrombosis  time in NSK 8 g/kg and 16 g/kg groups was significantly increased (P<0.05 or  P<0.01).Compared with blood stasis model group, the thrombus length was shortened and the dry weight and wet weight were reduced (P<0.05 or  P<0.01) in NSK 8 g/kg and 16 g/kg groups;the platelet aggregation rate (P<0.05) and whole blood and plasma viscosities were decreased (P<0.05 or  P<0.01) as well,the coagulation time was prolonged(P<0.05). Conclusion  NSK capsule can prevent thrombosis and improve the characteristics of  hemorheology in rats.

Abstract:Objective To investigate the association between MBD3 gene polymorphism and schizophrenia and provide theoretical basis for its etiological study. Methods The method of PCR-based ligase detection reaction (PCR-LDR) was applied to　detect the genotypes of the tag SNP on MBD3 gene in 50 family trios consisting of fathers, mothers, and affected offsprings with schizophrenia. Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness-of-fit χ test. The frequencies of genotype and allele of rs4807934 and rs4807122 locus of MBD3 gene were statistically computed. Results The distribution of genotypic frequency of rs4807934 and rs4807122 did not deviate from  Hardy-Weinberg equilibrium either in the patient group or in the parent group（P>0.05）.The haplotype relative risk test (HRR) showed there was no significant difference of frequency distribution of allele of rs4807934 and rs4807122 between patient group and parent group (P>0.05).The transmission disquilibrium test (TDT) showed the allele of rs4807934 and rs4807122 transmitted from heterozygotic parents to affected offsprings did not bias from 50 % (P>0.05) Conclusion The polymorphism of rs4807934 and rs4807112 of MBD3 gene may be not involved in the cause of schizophrenia in North Chinese Han population.

Abstract:Objective To investigate the relationship between the proximal femoral nail (PFN) fixation and postoperative anemia in the elderly patients with intertrochanteric fractures , and clarify whether the elderly PFN fixation of intertrochanteric fractures is the main risk factor  for postoperative anemia.Methods From January 2006 to August 2009, 43 patients with intertrochanteric fractures were selected( PFN treatment group), aged 71-95 years, with an average age of 82 years, 23 male and 20 female, according to Evans classification:ⅢA 20 cases, ⅢB 17 cases, Ⅳ type 6 cases;cause of injuries:25 cases of slip and fall ,15 cases of traffic accident fall ,3 cases of other diseases induced fall in a fainted.Control group, 19 cases, aged 70-85 years, mean 80 years old, 11 males and 8 females;according to Evans classification:ⅢA 9 cases, ⅢB 6 cases, Ⅳ type 5 cases;cause of injuries:11 cases of slip and fall, 7 cases of　traffic accident fall, 1 case of other diseases induced fall in a fainted.The number of red blood cells(RBC), hemoglobin(HB) and hematocrit(HCT) in PFN treatment group before and after operation were compared with control group.Results The number of preoperative RBC, HB, HCT before operation in PFN treatment group had no significant difference compared with control group (P> 0.05);the number of RBC, HB, HCT 2 and 3 d after operation in PFN treatment group were significantly decreased compared with before surgery(P<0.05).The number of RBC, HB, HCT 1 week after admission in control group were lower compared with admission,but there was no significant difference (P> 0.05).Conclusion The internal fixation of PFN in elderly patients with intertrochanteric fracture can lead to postoperative anemia, and recessive loss of blood is the main reason for postoperative anemia.

Abstract:Objective To evaluate the relationship between the polymorphism of lipoprotein lipase(LPL) PvuⅡP-P-genetype and coronary heart disease (CHD) in Chinese Han population,and expound the function of LPL PvuⅡP-P-genetype in occurrence and development of CHD.Methods According to the requirements of Cochrane systematic review,a thorough literature search about studies on the relationship between the polymorphism of LPL PvuⅡ and CHD was performed among PubMed,Medline,Chinese periodical full text database,Wanfang datebase and Vip information database.The qualities of documents were evaluated.RevMan 5.0 software was used to perform the Meta-analysis on those valid studies.Results Six literatures met the inclusion criteria,including 593 patients and 572 controls.The distribution of LPL PvuⅡgenotype of both groups in every study was in accordance with Hardy-Weinberg equilibrium.No heterogeneity was found between the six studies,nor the statistical publication bias.With the M-H mathematical model,the pooled odds ratio for PvuⅡP-P-/(P-P+/P+P+) in CHD group against control group was 2.32,95%CI(1.64,3.28),P<0.05.The funnel plots was symmetrical,which means there was no publish bias between the six studies.Conclusion Chinese Han population who carries the LPL P-P-genotype has a higher risk to be subjects of CHD,which suggests that the LPL P-P-genotype may be the susceptibility gene of CHD.

Abstract:Objective To study the chinical characteritics of critical patients cased by  influenza A H1N1 virus,a new pathogen,in order to provide references for diagnosis and treatment.Methods The general clinical data, symptoms, signs, laboratory examinations, diagnosis and treatment methods and results in 24 cases of influenza A H1N1  complicated with severe pneumonia   were analyzed. Results The mean age  of 24 patients was (28.73±9.24) years, in which 2 cases of pregnant women (8.3%), obesity (body mass index (BMI) ≥ 30) 18 cases (75.0%), chronic underlying diseases  2 cases (8.3 %), including 1 case of bronchial asthma, 1 case of post renal transplantation.Fever, sore throat, shortness of breath were the main symptoms of patients. Multiple lung consolidation was found in 22 cases (91.6%). 24 patients were  treated with anti-viral therapy (oseltamivir 75 or 150 mg,oral administration, 2 times　per day), 20 cases with  hormone therapy (83.3%), 18 patients were treated with mechanical ventilation (75.0%), of which 6 cases with mechanical ventilation after endotracheal intubation (25.0%), 2 death (8.3%). Conclusion Influenza A H1N1  complicated with severe pneumonia  mainly occurs in young adults; the pregnant women, obesity, patients with chronic underlying diseases are easily complicated  with severe pneumonia after infection;the  antiviral drugs,  glucocorticoids adjuvant therapy and artificial ventilation should be used as soon as possible. Tension pneumothorax, multiple organ dysfunction and other related complications are the main causes of death.

Abstract:Objective To discuss the value of H-MRS in diagnosis of common intracranial tumors by analyzing the metabolism differences between common intracranial tumors and normal brain tissue,and those among different intracranial tumors with H-MRS.Methods H-MRS manifestations of 27 cases of Ⅰ-Ⅱ grade glioma,22 cases of Ⅲ-Ⅳ grade glioma,24 cases of metastasis and 21 cases of meningioma were analyzed retrospectively,the peak values of N-acetylaspartate(NAA),creatine(Cr),choline(Cho) and lactate(Lac) were detected,the ratios of NAA/Cr,Cho/Cr,and NAA/Cho were compared.Results There were no significant differences among common intracranial tumors in routine MRI examination,and the H-MRS  manifestations of intracranial tumors were obviously  different with that of normal brain tissue:NAA was decreased(P<0.01),Cho was increased(P<0.05),and the NAA/Cr ratio in the region of glioma enhancement was obviously different with those of meningioma and metastasis (P<0.05).Conclusion H-MRS could distinguish differen kinds of intracranial tumors in molecular level,so it can make up  the defect of routine MRI.H-MRS can provide useful image information in intracranial tumors for pathologic classification and regulating  reasonable treatment principles.