Abstract:Objective To observe the effects of estrogen on the behavior and the norepinephrine level in the habenular nucleus of depressive rats,and explore the mechanism of estrogen involved in the process of antidepression.Methods Thirty rats were chosen from forty male Wistar rats,which had the similar scores in open field test.The depression models of rats were established by chronic mild stress.Open field test and forced swimming test were performed to detect the behavioral changes between rats with ovary excision (OVX) and with estradiol-treated ovary excision (OV/ER).High performance liquid chromatography (HPLC) was used to observe the changes of norepinephrine level in habenular nucleus.Results After 21-day chronic mild stress,the open field test showed that the vertical motion and cross motion  in OV/ER group were increased(P<0.05) and the immobile time in OV/ER group was decreased(P<0.05),when compared with those in OVX group.Forced swimming test demonstrated that the climbing time in OV/ER group was higher than that in OVX group (P<0.05);while the immobile time in OV/ER group was obviously lower than that in OVX group (P<0.05).The norepinephrine level　 in the habenular nucleus in OV/ER group was obviously increased compared with OVX group (P<0.05).Conclusion The habenular nucleus is a target of estrogen in central nervous system,which can take part in the antidepressive effect induced by estrogen.

Abstract:Objective To investigate the effects of oxidative stress on the expressions of transforming growth factor-β1(TGF-β1) and transforming growth factor-β receptor Ⅰ (TGF-βRⅠ) in rat unilateral ureteral obstruction (UUO) models.
Methods Thirfty  Wistar rats were randomly assigned into three groups:SOR group (sham-operated group,n=7);UUO group (operation group,n=11);US group (spironolactone 50 mg•kg^-1•d^-1 by daily gastric gavage after UUO,n=12).All the rats were killed  14 d after surgery.Renal fibrosis was assessed by the determination of tissue hydroxyproline (HYP) content.Malondialdehyde (MDA),superoxide dismutase (SOD) as well as aldosterone (ALD) content were measured.Histological changes were observed by HE and Masson staining.Immunohistochemistry  was performed to detect    the expressions of TGF-β1 and TGF-βRⅠ.Western blotting was used the determine the expression of  TGF-βRⅠ protein.Results Compared with sham group,the ALD contents in plasma and kidney tissues in UUO group significantly increased(P<0.01);the HYP and MDA levels in kidney tissues were also significantly increased(P<0.01),the SOD level was decreased(P<0.01);the expression levels of TGF-β1 and TGF-βRⅠ in kidney tissues were markedly increased(P<0.05).Compared with UUO group,the ALD contents in plasma and kidney tissues in US group were significantly decreased(P<0.01),the HYP and MDA levels in kidney tissues were also significanlty decreased (P<0.05，P<0.01)，the SOD level was increased(P<0.05)，the expressions TGF-β1 and  TGF-βRⅠ  in kidney tissues were markedly decreased(P<0.05). Conclusion The renal tubulointerstitial fibrosis induced by UUO may be related to the elevation of aldosterone,the increased production of oxidative stress and downregulation of TGF-β1 and TGF-βRⅠ.Aldosterone receptor antagonist spironolactone can partly alleviate renal fibrosis.

Abstract:Objective To investigate the internal fixation effects of poly(L-lactide) (PLLA) miniplates on the mandibular fracture in dogs.Methods A total of 12 dogs were involved in the study.After mandibular fracture models were created on both sides,one side of the mandibular was fixed with PLLA miniplates and the other side was fixed with titanium plates as control.The dogs were sacrificed separately at 1,3,6,12 months after operation,three animals at each time.Gross observation,radiography and histopathological examination were performed.Results All of the dogs were alive and fracture fixation of both sides was excellent after the operation.Gross observation:in 1 month group,the fracture line and some bony callus could be seen on both sides;in 3 months group,both sides had become into bone union;in 6 months group,both sides had become into bone union completely;in 12 months group,the fracture lines on both sides were difficult to be seen.Radiography examination:in 1 month group,the fracture lines and the screw holes could be seen on both sides;in 3 months group,the fracture lines turned ambiguity;in 6 months group,the line on the experimental side was difficult to be seen but the line on the control side disappeared;in 12 months group,the fracture lines on both sides disappeared.Histopathology examination:in 1 month group,there were lots of collagen fibers and a little bone trabecula on both sides;in 3 months group,the trabecula on the experimental side were much more than the other side;in 6 months group,the bone trabecula on both sides looked  like the normal bone tissue;in 12 months group,the bone trabecula became into nomal tissue.Conclusion The effect of PLLA miniplates for internal fixation of mandibular fracture is safety,the PLLA miniplate can maintain the stability of the bone fracture during the healing process.

Abstract:Objective To study the inhibition of  shRNA on  JNK expression  and influence on migration and invasion of mouse hepatocellular carcinoma cell line Hca-F  JNK and  discuss the correlation between expressing level of JNK and lymphatic metastasis of mouse hepatocarcinoma.Methods Three shRNA expression vectors were built and transfected to Hca-F cells stablely.　The inhibition of  shRNA on the expressions of mRNA and protein of JNK  were detected by RT-PCR and Western blotting.The most effective shRNA was selected.Transwell assay was used to detect the inhibition of shRNA on migration and invasion.Results The expression vector of pSilencer-shRNA was built and transfected to Hca-F cells successfully and the most effective shRNA was selected which inhibited JNK expression.The expression of JNK mRNA was markedly decreased after transfection of shRNA  ( P<0.01 ).The expression of JNK protein was markedly inhibited compared with the other groups ( P<0.01 ).The abilities of migration and invasion were decreased after transfection of shRNA ( P<0.05 ).Conclusion The Hca-F cell lines with markedly decreased expression of JNK by stable transfection is obtained by establishing pSilencer-shRNA vector.The inhibition of JNK expression could decrease the abilities of migration and invasion of mouse hepatocellular carcinoma cell lines.JNK maybe play an important role in lymphatic metastasis of hepatocellular carcinoma.

Abstract:Objective To construct the recombinant plasmid pCMV-Myc-PIASXβ and express the fusion protein in mammalian cells.Methods  PIASxβ fragment was digested from the original vector pGADT7 with SalⅠand NotⅠ,and then was inserted into the targeted pCMV-Myc vector by the recombinant DNA technique.After identification,the recombinant plasmid was transfected　into CHO cells.The expression of recombinant Myc-PIASxβ fusion protein was detected by Western blotting.
Results By the restriction enzyme digestion,fragment purification,ligation and transformation,the recombinant plasmid was obtained.The right recombinant plasmid pCMV-Myc-PIAS3 was identified with enzyme digestion and sequencing.By EcoRⅠ digestion analysis,pCMV-Myc-PIASxβ showed a 5641 bp band.By XbaⅠdigestion analysis,pCMV-Myc-PIASxβ showed two expected band of 3291 bp and 2349 bp.A specific protein expression band at 68 000 (PIASxβ fusion protein) was showed in Western blotting,which matched recombinant plasmid.
Conclusion The recombinant plasmid of pCMV-Myc-PIASxβ is sucssesefully constructed,which provids a good tool for further function study on PIAS family．

Abstract:Objective To observe the structure changes of the synapse of the neurons in hippocampus CA3 of young  rats and study the basis for the mechanism of polyene phosphatidyl in providing learning and memory and the effect on  synaptic plasticity. Methods A total of 20 Wistar rats with 5 months were randomly divided into polyene phosphatidyl group and normal control group. Each group had 10 rats. After 4 weeks feeding, Water maze training was peformed in all the rats for 1 weeks. The immue expressions of synapsin (SYN) of the rats in polyene phosphatidyl group  and  control groups  were observed with immunohistochemical method and analyzed by MOTAC imagine analyzing system. The change of synapse of hippocamal CA3 was observed with electron microscope. And  the other 24 to 26 months rats were selected as aged group,and  fed in the same condition. Moreover, the ultrastructures of hippocamal CA3 of aged rats were observed by transmission electron microscopy(TEM).Results The SYN in  polyene phosphatidyl group(0.430 0±0.022 4) was higher than that in  control group(0.3567±0.0209)  (P<0.05).  The density of SYN in polyene phosphatidy1 group was (102.69±8.96)•μm^-3 and the area of SYN membrane in the region of hippocamal CA3 was (0.066±0.010) μm²/μm³, which were higher than those in  control group(82.89•μm^-3±6.52•μm^-3, .046μm²/μm³±0.006 μm²/μm³ ) (P<0.05). Under TEM the  abundance of synapse had complete structure, the synapse cleft were clear and the spine apparatus were active. The synaptic vesicles in aged rats increased, the size was different, and accumulation presented. The spine apparatus were far away from postsynaptic membrane; the flat vesicles of the spine apparatus were expanded and increased in number.Conclusion Additive polyene phosphatidyl  can improve the expression  of SYN  in the region of hippocamal CA3 and  increase the density of SYN and the area of SYN membrane,  and improve  the special discrimination of learning and memory in young rats.

Abstract:Objective To investigate the expression and distribution of cyclooxygenase-2 (COX-2) in periodontal tissues during experimental tooth movement in rats,and  study  the relationship between  COX-2 and vesscular reconstruction in experimental tooth movement process.Methods Thirty-five healthy Wistar rats were divided randomly into 7 groups on average:normal group and experimental groups for 1,3,5,7,14 and 21 d.A NiTi coil spring with 0.294 N mesial force was connected between first molars of maxillary and the upper incisors.The histological sections were stained with goat anti rat COX-2 antibody,and computer image analysis was used to study the expression of COX-2 in the periodontal tissues of rats.Results Pressure area:compared with normal group(134.75±5.25) the COX-2 expression in 1 d group (147.73±3.27)increased (P<0.05) after tooth movement and reached the maximum at 5 d(154.32±9.54).Tension area:compared with normal group,the COX-2 expression in 3 d group (139.16±5.01)increased( P<0.05) after tooth movement and reached the maximum at  7 d(159.46±7.48).There was no significant difference of COX-2 expression in pressure area and tension area bewteen 21 d group and normal group (P>0.05).
Conclusion The expression of COX-2 in  periodontal tissues during experimental  tooth movement increase,suggesting that COX-2 can promote the vascular reconstruction in periodontal tissues during orthodontic tooth movement.

Abstract:Objective To investigate the effects of sphingosine-1-phosphate(S1P) on single-aisle delayed rectifier K⁺ current in heart ventricular myocytes of guinea pigs and explore the role of S1P in ventricular arrhythmia.Methods The Langendorff perfusion method was used to isolate ventricular myocytes of guinea pigs and the ventricular myocytes were randomly divided into normal control group,S1P(2.2 μmol·L^-1) group and S1P(2.2 μmol·L^-1)+Suramin（200 μmol·L^-1）group.The current of delayed rectifier potassium channel was recorded by cell-attached recording.Results Compared with normal control group by cell-attached recording and respecting the channel dynamics,the opening dwell time and the opening probability decreased,while the closing dwell time increased in S1P group (P<0.05)；and there was no obviously difference between S1P+Suramin group and normal control group while respecting the channel dynamics（P>0.05）.Conclusion S1P inhibits delayed rectifier potassium current of ventricular myocytes of guinea pigs by shortening its opening dwell time,extending its closing dwell time and reducing the opening probability.And the functions above are mediated by membrane receptor.

Abstract:Objective To construct secreting type human TRAIL(shTRAIL) gene vector pcDNA3.1-HRE/Egr1-shTRAIL mediated by hypoxia/radiation double sensitive promoter,and  observe the effect of  hypoxia and radiation on shTRAIL. Methods HRE upper and lower strands  were gotten by chemical synthesis,double strands HRE was gotten by PCR;pMD19T-Egr1 was digested by SacⅠ and Hind Ⅲ,then Egr1 was obtained,pshuttle-shTRAIL was digested by Kpn Ⅰ and BamH Ⅰ,then shTRAIL was obtained;HRE/Egr1 double sensitive promoter mediated shTRAIL expression vector pcDNA3.1-HRE/Egr1-shTRAIL was constructed by gene recombination technique,it was identified correctlly by enzyme digestion,PCR and sequencing.A549 cells were divided into normal,hypoxia(0.1%),irradiation(6 Gy) and hypoxia + irradiation groups.Results After enzyme digestion by BamH Ⅰ and Sma Ⅰ,the fragments which lengthes were 1284 bp and 4 998 bp,2 292 bp and 3 990 bp were obtained;the vector was amplified by PCR with Egr1 and shTRAIL primer,the products which lengthes were 469 bp and 820 bp were obtained;pcDNA3.1-HRE/Egr1-shTRAIL was sequenced,the result was same to designed,this demonstrated that the construction was right.The vectors were transfected into A549 cells of adenocarcinoma of lung,the expression levels of  shTRAIL mRNA and protein were increased after treated with hypoxia and radiation,it had statistically significant differences compared with normal group (P<0.05),and when they  were combinated,the effect was more obvious.Conclusion Secreting type human TRAIL gene vector pcDNA3.1-HRE/Egr1-shTRAIL mediated by hypoxia/radiation double sensitive promoter is constructed successfully,and hypoxia and radiation could increase the  expression of TRAIL,and they have synergetic effect.

Abstract:Objective To observe the effect  of fluvastatin (Flu) on differentiation and induction of human promyelocytic leukemia HL-60 cells and provide the  theoretical  foundation for treatment of human promyelocytic leukemia. Methods The cultured HL-60 cells were treated with  20 μmol·L^-1 Flu,the morphological changes  of the cell differentiation were examined.The NBT reduction capability was detected in HL-60 cells treated with Flu for 72 h.After HL-60 cells were treated with 20  μmol·L^-1 Flu for 5 d,  they were stained with non-specific esterase and the percentage of differentiation cells was analyzed.Results The HL-60 cells treated with Flu showed smaller cell body,reducing proportion of nucleus to cytoplasm,the nucleus tortuosity,fold or sublobe.There were specific granules and vacuoles in cytoplasm,displaying that some cells had differentiated to relative mature cells.As compared with control group,the NBT reduction capability in HL-60 cells treated with Flu for 72 h was significantly higher than that in control cells (P<0.01),which was similar to that of ATRA (10  μmol·L^-1.After HL-60 cells were treated with 20  μmol·L^-1Flu for 5 d,the percentages of positive cells were markedly higher than those in untreated HL-60 cells,and the positive reaction was not inhibited by NaF.Conclusion Flu treatment can result in differentiation of HL-60 cells to relative mature cells,it indicates that Flu may be used as a drug for treatment of human promyelocytic leukemia.

Abstract:Objective To investigate the construction of compound membrane with corneal stromal cells and collagen-chitosan by tissue engineering technique and its biocompatibility.
Methods Rabbit and human corneal stromal cells were separated and seeded into collagen-chitosan membrane.The compound membrane was transplanted into rabbit corneal stroma.Then the  growth condition of keratocytes,the effect on normal keratocytes and degradation of compound membrane were detected by corneal confocal microscope,anterior OCT and  histological and immunohistochemical methods ex vivo 1,2,4 weeks after grafting.Results The rabbit and human corneal stromal cells grown well  in collagen-chitosan scaffold.The compound membrane degradated gradually after grafting.There was no necrosis and dissolvation.Corneal epithelium,stroma and endothelial cells were all normal.Conclusion Collagen-chitosan can be used as a biological scaffold for construction of corneal stroma.Corneal confocal microscopy and anterior OCT are new methods to observe the biological activity of constructed corneal stroma.

Abstract:Objective To study the effect of the mitochondrial oxidative lesion in the pathologic mechanism of vascular dementia(VD) in rats,and provide theoretical basis for treatment of VD.Methods The VD rat model through ligating bilateral occlusion of both common carotid arteries was made.The rats ware divided into sham group,ischemia-1-month group,and ischemia-2-month group.The pathologic   and histologic changes of rats after Morris Water Maze and place navigation were determined.The SOD activity and  the MDA content were measured by UV-spectrophotometer and the fluidity of mitochondrial membrane was detected by fluorescence spectrophotometer.
Results Compared with sham group,the escape latencies in ischemia-1-month and ischemia-2-month group were prolonged significantly(P＜0.05)，the SOD activity was decreased significantly(P＜0.05),the MDA content and the viscous coefficient (η) of mitochondrial membrane were significantly increased(P＜0.05).
Conclusion The pathologic mechanism in VD rats is concerned with the metabolism of SOD and MDA and the fluidity of mitochondrial membrane,and is  related to the mitochondrial oxidative stress.Above all,mitochondrial oxidative lesion may be one of pathologic mechanism of VD.

Abstract:Objective To study the effects of formaldehyde on apoptosis and cell cycle of testicle cells in male mice,and explore the mechanism of formaldehyde on reproduction and genetic toxicity of male mice.Methods The male Kunming mice were randomly divided into negative control group (NC),formaldehyde exposed groups (21,42,84 mg·m^-3 )　 and positive control group (PC).12 male mice were included in each group.The exposed dose was respectively 21 mg·m^-3  (1/24LC₅₀),42 mg·m^-3  (1/12 LC₅₀) and 84 mg·m^-3  (1/6 LC₅₀).Mice in negative control group inhaled air,the mice in positive control group were injected with cyclophosphamide.The mice were killed after exposure to toxicant,and the testicle cell apoptosis and cell cycle in mice in various groups were examined.Results The apoptotic rate of mouse testicle cells in 1/6LC₅₀ group was significantly higher than that in negative control (P<0.05).The percentages of S phase cells gradually increased and that of G₀/G₁ and G2+M　phase cells gradually decreased with the formaldehyde dose increasing.The percentages of S-phase cells in three exposed groups were obviously higher than that in negative control and that ofG0/G1 and G2+M　phase cells in 1/6 LC₅₀ group were obviously lower than other groups(P<0.05).Conclusion Formaldehyde can increase the apoptotic rate of mouse testicle cells,and  increase the percentages of S phase cells and decrease the percentages  of G0/G1and G2+M phase cells.The results suggest that formaldehyde has reproductive and genetic toxicity.

Abstract:Objective To study the injury of the neural stem cells cultivated in vitro induced by glutamate of embryonic rats and provide theoretical basis for development of protective drugs of neural stem cells.
Methods The Wistar rat(15 d of gestation) embryonic brain tissues were cultivated  in vitro and the cells were confirmed as the neural stem cells with immunohistochemical method.The neural stem cells cultured  normally were used as control,the different concentrations of glutamate were added,then the survival rate of neural stem cells was observed with MTT colorimetry.Results The differentiation cells from neural stem cells had both neurons and neuroglial cells that presented neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP )dying masculine respectively.After treated with glutamate for 24 h,the survival rates of neural stem cells in glutamate groups were obviously reduced compared with control group.The survival rate in 50 μmol·L^-1  glutamate group was 86.45%,compared with control group,there was no significant difference (P>0.05).The survival rates of the stem cells in 100  and 1 000 μmol·L^-1  glutamate groups (71.22%，15.14%) were significantly lower than that in control group (P<0.05，P<0.01).Conclusion Glutamate has function of damaging on the neural stem cells in a dose-dependent manner;when  the dose of  glutamate  increases,the survival rate of the neural stem cells  decreases gradually.

Abstract:Objective To study the anti-tumor effect of aspirin and celecoxib on breast cancer cells MCF-7 through investigating their effects on the expressions of COX-2,VEGF and proliferation of MCF-7 cells. Methods The human breast cancer cells MCF-7 were divided into 2.5,5.0 and 10.0 mmol·L^-1aspirin groups and 30,60 and 120 μmol·L^-1celecoxib groups,the MCF-7 cells without treatment were used as negative control group.The inhibitory rates were detected by MTT assay after MCF-7 cells were treated for 24,48,72 h. The expressions of COX-2 and VEGF in MCF-7 cells after treated for 24 and 48h were detected by immunohistochemical assay.
Results  ①　There were no significant differences of the inhibitory rates of the MCF-7 cell line between 2.5 mmol·L^-1 aspirin group(48 h),30 μmol·L^-1celecoxib group (48 and 72 h),60 μmol·L^-1celecoxib group (48 h) and control group(P>0.05),the inhibitory rates in the other groups were increased compared with control group(P<0.05). The inhibitory rates of MCF-7 cells in 10.0 mmol·L^-1 aspirin group after treated for 24,48 and 72 h were higher than those of 2.5 mmol·L^-1aspirin group at the same time (P<0.05). The inhibitory rates of MCF-7 cells in 120 μmol·L^-1 celecoxib group after treated for 24,48 and 72 h were higher than those in 30 μmol·L^-1or 60 μmol·L^-1 celecoxib groups at the same time(P<0.05).The inhibitory rates of MCF-7 cells in 2.5,5.0 and 10.0 mmol?L^-1  aspirin groups after treated for 72 h were higher than those of MCF-7 cells after treated for 24 h(P<0.05).②The expressions of COX-2 and VEGF located in MCF-7 cytoplasm with yellow and pale brown staining.③There were no significant differences of the expressions of COX-2 and VEGF in MCF-7 cells between 30 μmol?L^-1 celecoxib group(24,48 h),60 μmol·L^-1celecoxib group(48 h) and negative control group(P>0.05),the expressions of COX-2 and VEGF in MCF-7 cells in the other groups were lower than that in negative control group(P<0.05).The expressions of COX-2 and VEGF in MCF-7 cells treated with 10.0 mmol·L^-1aspirin for 24 and 48 h were decreased compared with those in 2.5 or 5.0  mmol·L^-1aspirin groups at the same time (P<0.05).The expressions of COX-2 and VEGF in MCF-7 cells treated with 120 μmol·L^-1 celecoxib for 24 and 48 h were decreased compared with those in 30 μmol·L^-1 celecoxib group at the same time(P<0.05). The expressions of COX-2 and VEGF in MCF-7 cells treated with 10.0 mmol·L^-1aspirin or 120 μmol·L^-1celecoxib for 48 h were decreased compared with those of MCF-7 cells treated for 24 h(P<0.05).
Conclusion Aspirin and celecoxib have inhibitory effects on the growth of human breast cancer cells MCF-7,and they can inhibit the expressions of COX-2 and VEGF in MCF-7 cells in a dose and time dependent manner.

Abstract:Objective To investigate the effect of tripterygium wilfordii hoff-F on acute immune rejection of bone xenograft in mice.Methods Male Neijiang swine and BALB/c mice were used as donor and receipts,respectively.Forty mice were divided into experimental group and control group at random.The mice in experimental group were administered intragastrially with tripterygium wilfordii hoof-F before bone xenograft.At two weeks postoperatively,CD4⁺,CD8⁺,CD4⁺/CD8⁺T lymphocytes in spleen were detected by flow cytometry,and the levels of IL-2,IFN-γ and IL-4 in blood were analyzed by quantitative ELISA assay.And the infiltration of lymphocytes and macrophages surrounding soft tissues of bone graft was also observed in order to evaluate the immunosuppression of tripterygium wilfordii hoof-F.
Results At two weeks postoperatively,compared with control group,CD4⁺,CD8⁺,CD4⁺/CD8⁺T lymphocytes in spleen,the levels of IL-2,IFN-γ and IL-4 in blood of mice in  experimental group were significantly decreased(P<0.05).There was few lymphocytes infiltration surrounding soft tissue of bone graft in experimental group.

Abstract:Objective To investigate the protective effect of Schizandrae Lignanoid(SCL）on the apoptosis of PC12 cells induced by  H₂O₂ and its relative mechanisms.  Methods PC12 cells were divided into four groups： control group,model group,high dose SCL (SCL₁) group， and low dose SCL(SCL₂) group.Apoptosis of PC12 cells was induced by  H₂O₂.The cell activity was determined by MTT,the cell apoptotic rate was determined by Annexin V-FITC/PI and ΔΨm was detected by flow cytometry.The expressions of bcl-2 and bax were detected by immunohistochemistry. Results Compared with model group，SCL increased  the survival rate of PC12 cells (P<0.01),decreased the apoptotic rate of PC12 cells  (P<0.01) and protein expression of bax (P<0.05),and increased the △Ψm  (P<0.01) and protein expression of bcl-2 significantly (P<0.05)  in a dose-dependent manner.Conclusion SCL can inhibit the apoptosis of PC12  cells induced by H₂O₂ ,which might be correlated with the inhibition of apoptosis in the path way of mitochondrion.Co

Abstract:Objective To  study the effects of lactate peritoneal dialysis solution(L-PDS) with different concentrations on apoptosis of human peritoneal mesothelial cells(HPMC),the expressions of bcl-2,bax and activity of caspase-3.Methods HPMC were separated using enzyme digestion and cultivated stably in vitro.After HPMC were co-cultivated with different concentrations(1.50%,2.50%,4.25%) L-PDS,flow cytometry  was used to test the apoptosis  of HPMC,RT-PCR was used to observe the expressions of bcl-2 and bax,fluorometric method was used to detect the activity of caspase-3.
Results Compared with control group, L-PDS could induce the apoptosis of HPMC,especially in high concentration.With the increasing of  L-PDS concentration,the expression of bcl-2 mRNA decreased,the expression of bax mRNA increased,the activity of caspase-3 raised.There were significant differences of the indexes mentioned above between 4.25%,  2.50% L-PDS groups and  control group（P<0.05），there also was significant difference between 4.25% L-PDS group and  2.50% L-PDS group(P<0.05)，but there was  no significant difference  between 1.5% L-PDS group and  control group(P>0.05).
Conclusion L-PDS could induce HPMC apopto9sis,which may be executed by alternating of the expressions of bcl-2,bax and activating of caspase-3.

Abstract:Objective To study the cardiac damage of the heroin dependence and withdrawal rats and the influence of myocardial enzymes by purine nucleotide.Methods The rat models of heroin dependence and withdrawal were set up.80 Wistar rats were randomly divided into 8 groups(10 rats of each group):control(C),heroin administration group(H),3 d withdrawal group(W3),9 d withdrawal group(W9),nucleotide group(N),heroin and nucleotide group(HN),nucleotide 3 d group(N3),nucleotide 9 d group(N9).The activities of total creatine kinase(CK),the myocardial isoenzyme of creatine kinase(CK-MB),lactate dehydrogenase (LDH),a-hydroxybutyrate dehydrogenase (HBDH) and m-aspartate aminotransferase (m-AST) in plasma were tested.
Results Compared with control group,the activities of CK,CK-MB,LDH,HBDH and m-AST in 3 d withdrawal group and 9 d withdrawal group were significantly lower (P<0.01,P<0.05),while the activities of  CK,CK-MB,LDH,HBDH and m-AST in heroin administration group,nucleotide group,heroin and nucleotide group didn’t change significantly (P>0.05).And compared with control group,the activities of HBDH in 3 d nucleotide group and 9 d nucleotide group increased significantly (P<0.05,P<0.01),and the activities of CK,CK-MB,LDH and m-AST were considered normal (P>0.05).
Conclusion The heroin has delayed and durative damage to the cardiac muscle,and can’t eliminate immediately after stopping administration；purine nucleiotide can diminish this damage and maintain some plasma myocardial enzymes of heroin dependence  and withdrawal rats in relatively steady condition，and improve the functional status of cardiac muscle cells.

Abstract:Objective To investigate the effect of cellular adhesion tripeptide  RGD on the growth and proliferation of human stomach cancer cells BGC823,and study the function of cellular adhesion tripeptide RGD in inducing BGC823 apoptosis.
Methods BGC823 cells cultivated in vitro were divided into control group,12.5,25.0,50.0 and 100.0 mg?L^-1  RGD groups. BGC823 cells were treated with cellular adhesion tripeptide RGD at various concentrations. MTT assay was used to examine the growth and proliferation of BGC823 cells after treatment with cellular adhesion tripeptide RGD for 48 h.Ordinary light microscope and transmission electron microscope were used to observe the morphology of apoptotic cells,and immunohistochemical method,flow cytometry were also used to analyze the BGC823 apoptosis.Results The results of MTT assay showed that in 12.5,25.0,50.0,100.0 mg·L^-1RGD groups after treated for 24,48 and 72 h,the inhibitory rates of  BGC823 were signiticantly higher then those in cortrol group  (P<0.01);with the increasing of the concentration of RGD,the inhibitory rate of  BGC823 cells increased.The results of ordinary light microscope showed that after treated with 50.0 mg·L^-1 RGD for 48 h,the morphology of BGC823 cells had some transmutations,the cells became smaller,nucleus was deeply stained,cytoplasm was red,and a lot of apoptotic cells were observed;the results of transmission electron microscopy observation  showed that the cell surface microvilli disappeared,and heterochromatin was aggregated toward the edge,they were all gathered on the nuclear membrane,formed a crescent-shaped cap structure,and the nuclear membrane was even disappeared,nucleus was fragmented,and  the  apoptotic bodies could be seen;the results of flow cytometry showed that BGC823 cells had undergone apoptosis,and the apoptotic peak became higher as the concentration increased;the electrophoresis results showed that the BGC823 cell’s DNA fragments had broken into various small fragments,limpid degradation of electrophoresis strips were observed.The result of immunohistochemistry showed that after  BGC823 cells were treated with 50.0 mg·L^-1  RGD,the Survivin expression was significantly  diminished,the average gray values were 196.56 ± 10.12 and 108.59 ± 14.26 respectively in control group and experimental group according to the results of the image analysis,there was significant difference(P<0.01),which indicated the apoptosis of BGC823 cells in experrmental group.
Conclusion Cellular　 adhesion tripeptide RGD may inhibit tumor cell growth and proliferation through inducing apoptosis,and the mechanism of inducing BGC823 apoptosis  is probably initiated mitochondria into this category.

Abstract:Objective To study the effects of eukaryotic expressive mature peptide LL-37 of human cationic antimicrobial peptide (hCAP-18) on the expressions of membrane molecules on dendritic cells (DCs). Methods By gene cloning,the eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 for the mature peptide LL-37 of hCAP-18 was constructed.Then they were transfected into HEK293 cell lines.After the cell lines were cultivated for 48 h，the supernatant was collected.Then the DCs from peripheral blood mononuclear cells (PBMCs) induced by rh-GM-CSF and rh-IL-4 were cultivated with the supernatant for 48h.The expressions of membrane molecules CD40 and HLA-DR on DCs were detected by flow cytometry (FCM).
Results  The eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 was constructed successfully and it expressed in eukaryotic cells HEK293.FCM results indicated that the expressions of CD40 and HLA-DR on the membrane of DCs which were stimulated by the supernatant produced by pcDNA4/Myc-His-LL-37 were higher than those in  control group(P<0.05). Conclusion The mature peptide LL-37 of hCAP-18 is constructed successfully and it can increase the expressions of the  membrane molecules on DCs such as CD40 and HLA-DR.

Abstract:Objective To investigate the effects of nerve growth factor (NGF) on metabolism and function  of neuroglial cells in the normal mice and the nonobese diabetic (NOD) mice.Methods  The mice were  divided into normol group and NOD group (n=10).The neuroglial cells  abstracted from the normal mice and the NOD mice were randomly divided into control  (with RPMI-1640) and 4 experimental groups (n=10);5,10 and 20 μg·L^-1NGF and 20 μg·L^-1LPS +10 μg·L^-1NGF were used in experimental groups.The activity of superoxide dismutase(SOD) and malonic dialdehyde(MDA) content in the culture solution were detected with xanthine oxidase and TBA methods.
Results Compared with control group of normal mice,the  activity of SOD and MDA content in neuroglial cells  of the NOD mice were significantly　decreased (P<0.01)；compared with that of respective control group,the activities of SOD were increased (P<0.05)and the MDA contents were decreased  significantly (P<0.05) in    the NOD mice of NGF groups.
Conclusion  NGF has some effects on the improvement of cell metabolism and function in the NOD mice.

Abstract:Objective To observe the efficiency of vascular endothelial cell growth factor(VEGF) gene transferring into fibroblasts in vitro by diagnostic ultrasound combined contrast agent. Methods  The fibroblasts were divided into four groups:pure VEGF₁₆₅ plasmid,contrast agent and VEGF₁₆₅plasmid,ultrasound plus VEGF₁₆₅ plasmid (MI 1.6),ultrasound plus contrast agent and VEGF₁₆₅plasmid (MI 1.2,1.4,and 1.6). The transfection efficiencies of the VEGF gene in the fibroblasts were observed under confocal microscopy respectively in different groups.
Results There was no significant difference of transfection efficiency among ultrasound plus plasmid group,contrast agent plus plasmid group and pure plasmid group (P>0.05).The transfection efficiencies were improved significantly in the ultrasound plus contrast agent and VEGF plasmid group (MI 1.2,1.4,and 1.6) compared with pure plasmid group（P<0.01).The transfection efficiencies were higher in ultrasound plus contrast agent and VEGF₁₆₅ plasmid group (MI 1.4,1.6),but there was no significant difference between the two groups (P>0.05);there were significant differences between ultrasound plus contrast agent and VEGF₁₆₅plasmid(MI 1.4，1.6)　 and  ultrasound plus contrast agent and VEGF₁₆₅plasmid group (MI 1.2) (P<0.01).
Conclusion Ultrasound combined with contrast agent can improve the transfection efficiency of VEGF gene,and the transfection efficiency increases with the intensity of ultrasound.

Abstract:Objective To explore the anti-convulsion action of histamine(HA) in the central system and treatment of histamine H₃ receptor(H₃R) antagonists to rat model with intractable epilepsy.Methods 88 Wistar rats (12-day-old) were randomly divided into three groups:normal control group,N-methyl-D-aspartate(NMDA) group and betahistine(BH) groups (including high and low dose BH groups).Wistar rats  received an intraperitoneal NMDA administration to make animal model of intractable epilepsy at infant period and toddler age.After that,the rats were observed daily for latencies and incidences to two NMDA-dependent stereotypical behaviors.The HA content of each brain region was determined with fluorimetry,and  H₃R were evaluated with immunohistochemical  method.Results The automatisms including tail twisting and emprosthotonus seizures of (12-17)-day-old rats were observed in NMDA and BH groups.The  rats,aged 18-25 d,became quiet following automatisms rather than emprothotonic.Compared with NMDA group,BH groups had longer latencies and lower incidences of tail twisting and emprosthotonus(P<0.05).At the end of the 2nd week,compared with normal control group,the HA contents of cortex and hippocampus in NMDA group were lower(P<0.05),and the expression rates of H₃R positive cells were higher(P<0.05).Compared with NMDA group,the HA contents of cortex and hippocampus in BH groups were higher(P<0.05),the expression rates of H₃R positive cells were decreased(P<0.05).At the end of the 4th week,the changes of HA contents between every two groups became less.There were significant differences of the HA contents between NMDA group and BH groups(P<0.05).But there were no significant differences of the expression rates of H₃R positive cells between every two groups(P>0.05).Conclusion The NMDA-induced model is similar to the clinical manifest of human West syndrome.It is up to animal model of intractable epilepsy at infant period and toddler age.The HA content of brain region is negatively related with seizure incidence.H₃R antagonists have certain therapeutic function to intractable epilepsy in rats at infant period and toddler age.

To investigate the influence of genistein(Gen) on the growth  and apoptosis regulation of hepatoma carcinoma cells SMMC-7721.Methods SMMC-7721 cells were divided into three Gen groups treated with different concentrations(5.0,10.0,20.0 mg·L^-1）and control group(without Gen).After SMMC-7721 cells were treated for 24 and 48 h,the changes of ultrastructure of cells were observed under electron microscope, MTT was used to detect the inhibitory rate of proliferation of SMMC-7721 cells, flow cytometry was performed to analyze cell phase;immunohistochemistry was used to detect the expression of Caspases-3 protein.Results Compared　 with control group,chromatin in cellular nucleus became conglomeration or lumping,with chondriosome swelling in Gen groups. With the increasing of Gen concentration,the cellular nucleus became condensed,heterochromatin agglutinating,ground cytoplasm obviously cavitating;with the increase of Gen concentration and prolongation of  time,from 24 to 48 h, MTT showed the inhibitory rate of  proliferation  of SMMC-7721 cells  increased  in a time- and dose-dependent  manner.Flow cytometry analysis showed that with the increase of Gen concentration,the number of cells at G₂/M  phase increased,the number of cells at S phase decreased and the ratio of G₀/G₁ also decreased.Immunohistochemistry results showed the expression of protein Caspases-3 increased with the Gen concentration in a dose
-dependent manner（P<0.01）.
Conclusion Gen has significantly  inhibitory effect on the growth of hepatoma carcinoma cells SMMC-7721,up-regulating the expression of Caspases-3 protein and inducing  apoptosis may be one of the mechanisms.

To investigate the expression and correlation of Skp2 and P^27kip1 in colon carcinoma tissue and explore the relationship with the development of colon carcinoma.Methods  The protein expressions of Skp2,P^27kip1,P53 and Bcl-2 were detected by immunohistochemistry staining (SABC method) in 50 cases of colon carcinoma  tissue (CCT) and 20 normal colonic mucosas (NCM).The positive expression rates in different tissues were compared and the relationship between the expression and different clinicopathological parameters were analyzed.Results The expression of Skp2 in the CCT was significantly increased compared with NCM（P<0.05）.The protein expression of P^27kip1 in  CCT was significantly lower than that in NCM（P<0.05）.The expressions of Skp2 were significantly increased in the Duke’s staging C and D group,lower degree differentiation group and lymph node metastases positive group（P<0.05）.The expressions of P^27kip1 were significantly increased in the Duke’s staging A and B group,high degree differentiation group and  lymph
 node metastases negative group（P<0.05）.The expressions of P53 and Bcl-2 protein were significantly increased in the low degress differentiation CCT（P<0.05）. The negative correlation was present between  Skp2 and P^27kip1 in  CCT（r=－0.282，P<0.05）,and there were  positive correlations  between Skp2 and P53,Bcl-2  in  CCT (r=0.345，r=0.231,P<0.05）.Conclusion The abnormal expressions of both Skp2 and P^27kip1 participate in  the differentiation,infiltration and  metastases of colon carcinoma. It may be realized through  up-regulating the expressions of P53 and Bcl-2.

To investigate the association between the single nucleotide polymorphism(SNP) of interferon-gamma(IFN-γ) gene +874 site and susceptibility to ovarian cancer,and provide a theoretical basis for the gene diagnosis of ovarian cancer. Methods Using the PCR,cloning and sequencing techniques,the genotypes of IFN-γ gene +874 site A/T of 87 cases of ovarian cancer patients(case group) and 86 normal women(control group) were detected.Results The distribution of genotypic frequency did not deviate from Hard-Weinberg  equilibrium in both case group and control group;the genotypic frequencies of AA,AT and TT  had no significant difference between case group and control group (P>0.05);the frequencies of IFN-γ gene +874 site T and A allele showed significant difference between case group and control group (P<0.05),the relative risk suffering from T allele was 1.80 times of A allele;the relative risk suffering from TT genotype was 2.99 times of AA genotype;the genotypic frequency and allelic frequency had no significant difference between epithelial ovarian cancer and non-epithelial ovarian cancer in case group (P>0.05).Conclusion T allele of IFN-γ gene+874 site might have a relationship with generation of ovarian cancer,TT genotype might be susceptibility genotype for ovarian cancer.

To investigate the association between schizophrenia and the single nucleotide polymorphism（SNP） of  HIR histone cell cycle regulation defective homolog A  (HIRA).Methods PCR-restrictive fragment length polymorphism（PCR-RFLP） was used to examine the SNP of  rs1473109（A/G） of HIRA in 216 Han core family trios of schizophrenia.Data was analyzed using family based linkage disequilibrium.
Results The distribution of   allelic and genotypic  frequencies of rs1473109 of HIRA didn’t deviate from Hardy-Weinberg equilibrium.The result of TDT indicated that rs1473109 of HIRA gene had no association with schizophreaia.There were significant relationships between the distribution of genotypic frequency of rs1473109 of HIRA gene and delusion of observation（χ²=9.310，v=2,P=0.010）,experience of being revealed (χ²=14.373，v=2,P=0.001) and incoherence of thinking (χ²=11.067，v=2,P=0.004). Conclusion  As fa
r as Chinese Han people with schizophrenia,HIRA itself or gene adjacent to it may be associated with some of positive symptoms in schizophrenia.

To study the preemptive analgesia effects of flurbiprofen axetil on respiratory function in patients undergoing thoracotomy.Methods Twenty male patients undergoing thoracotomy were devided into two groups equally with ten cases each to receive either flurbiprofen axetil 5 mL (50 mg) (experiment  group) or saline 5mL(control group) 15 min before incision.At the end of the surgery both groups received intravenous sufentanyl PCA with a loading dose of 10  g,a maintenance dose of 2 g•h<^1,a bolus dose of 2 g and a lockout interval of 15  min.Age,weight,sex,history of general anaesthesia were recorded.Analgesia grades at 4，8，12，24，and 48 h after operation were accessed and  side effects(nausea and vomiting) were recorded.Total drug use,number of boluses delivered,number of boluses demanded were collected.Pulmonary function (FVC,FEV1,MMEF) was tested the day before operation,as well as at 24 and 48 h after operation.Results No differences between two groups were found for demographic data.There were also no clinically relevant differences between two groups with regard to side effects (P>0.05).Significant differences were observed in analgesia grade between two groups.Visual analogue scores (VAS) in different postoperative time points (4,8,12,24 and 48h) were lower in experiment group  than those in  control group (P<0.05).Also less bolus doses were demanded and delivered in experiment group (P<0.05).Respiratory function test in both groups was not as good as that before operation. 24 and 48 h after operation,the respiratory function test  was better in experiment  group  than that in control group, but there was no significant difference(P>0.05).Conclusion Flurbiprofen axeti1 has preemptive analgesia effects with the chosen dosage regimen in patients undergoing thoracotomy and doesn’t increase side effects.But the preemptive analgesia can’t improve postoperative respiratory function.

To investigate the relationship between the expressions of  P16 and proliferating cell nuclear antigen(PCNA) in osteosarcoma and clinicopathological features,and explore the effects of them in occurrence and development of osteosarcoma.
Methods The expressions of P16 and PCNA were detected by immunohistochemistry(SP) in 71 osteosarcima tissues and 10 normal bone tissues.
Results  ①The positive rate of P16 expression in osteosarcoma tissues was lower than that in normal bone tissues(P<0.05),the positive rate of PCNA expression in osteosarcoma tissues was higher that in normal bone tissues(P<0.05).②The expression of P16 was correlated with osteosarcima’s histological grade and prognosis(P<0.05),and wasn’t correalted with histological type and clinical stage(P>0.05).The expression of PCNA had positive relationship with osteosarcoma’s prognosis(P<0.05),and had no relationship with histological type and clinical stage and histological grade (P>0.05).③There was negative correlation between P16 and PCNA expressions(r^_s=－0.58,P<0.05).
Conclusion P16 and PCNA can play an important role in the occurreace,development and the metastasis of osteosarcoma,and have close relationship with its prognosis.

To investigate the relationship between plasminogen activatorinhibitor-1 (PAI-1) 4G/5G gene polymorphism and  homeostatic activation after percutaneous coronary intervention（PCI），and provide foundation for clinic  detection of thrombosis after PCI.Methods The plasma von Willebrand factor（vWF）,activated factor Ⅶ ( FⅦc) ,PAI-1 activities and GMP-140, D-dimers(D-D)  levels in 86 patients with coronary heart disease undergoing in-stent implantation were determined before and immediate postoperative,and 24 h after PCI.PAI-1 gene polymorphism was detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) to observe the changes of platelet function and coagulation and fibrinolytic states before and after PCI.Results The distribution of PAI-1 genotypes was as follows:48 cases with 4G/4G (56%),30 cases with 4G/5G (35% )and 8 cases with 5G/5G ( 9%).Among the patients,the allelic frequencies of 4G and 5G  were 73% and 27% respectively.No significant difference of the plasma GMP-140,vWF,FⅦa,D-D and PAI-1 levels was found between the three genotypes before PCI (P>0.05).In the patients with  4G/4G polymorphism of PAI-1 gene,the GMP-140,vWF,FⅦa,D-D and PAI-1 levels in plasma were significantly increased at  immediate postoperative period(P<0.01).The PAI-1 levels in plasma were significantly increased 24 h after PCI(P< 0.01);but the GMP-140,vWF,FⅦa and D-D  levels did not significantly increase (P>0.05).In the patients with 4G/5G and 5G/5G genotypes, all indexes mentioned above were significantly increased at  immediate postoperative period (P<0.01);24 h after PCI there were no significant differences (P>0.05).In the patients with 4G/4G genotype, the PAI-1 level in plasma was significantly higher than those in the patients with 4G/5G and 5G/5G genotypes 24 h after PCI(P< 0.01).
Conclusion There is a remarkable homeostatic activation and the fibrinolytic activity changes more in patients with 4G/4G genotype than  those in patients with  4G/5G and 5G/5G genotypes after PCI.The results suggest that the patients with 4G/4G genotype may be a dangerous factor in acute or subacute coronary thrombosis after PCI

To investigate the association between polymorphism of protein C inhibitor（PCI）gene G10877T and male infertility,and provide theoretical basis for treatment of male infertility.Methods PCR and sequencing technique were applied to detect PCI gene G10877 T polymorphism in 53 normal control and 102 male　 infertility.Results There were three genotypes of wild type（G/C）,hybridization mutation（G/T）and pure mutation （T/T）.The analysis of sequencing indicated that in sperums of a proportion of the male infertile patients,TGG in PCI gene G10877T mutated into TGT.The contrast of BLASTB indicated that this mutation made Trp in 271 position change into Cys.Compared with control group,TT genotypic frequency and T allelic  frequency in male infertility group had significant differences（P<0.05).
Conclusion
 PCI gene G10877 T polymorphism is related to  male infertility,and the  polymorphism of PCI gene increases the risk of male infertility morbidity.

To study the relationships between serum follicle stimulating hormone(FSH),luteinizing hormone (LH),testosterone (T),inhibin B(INHB) and sperm density and the significance of serum reproductive hormones in diagnosis of male infertility.Methods  150 patients with male infertility were selected,including 50 cases of oligospermia,50 cases of severe oligospermia,50 cases of azoospermia,and 50 healthy subjects were used as controls(normal sperm density group).Sperm density was detected by computer assisted sperm analysis(CASA).The serum FSH,LH,T,INHB levels were detected by radioimmunoassay.
Results The serum FSH and LH in oligospermia，severe oligospermia and azoospermia groups were significantly higher than those in normal sperm density group (P<0.01)，however the serum INH-B levels were significantly lower (P<0.01).The serum  FSH and LH levels in azoospermia group were significantly higher than those in oligospermia and severe oligospermia groups (P<0.01),while the serum INH-B level was significantly lower (P<0.01).There were significantly negative correlations between sperm density and the levels of FSH，LH (r=－0.562,P<0.001;r=－0.347,P<0.001). There was significantly positive correlation between sperm density and INHB (r=0.623,P<0.001),the sperm density had no correlation with T（r=0.080,P>0.05）.
Conclusion  There are negative correlations between the levels of FSH, LH and sperm density;and  the level of INHB is positively correlated  to the sperm density.

To research the geometrimorph of patello-femoral joint and the geometric correlation between its components by imageological methods and provide theoretical basis for total knee prosthesis design.Methods  The geometrimorph of patello-femoral joint of 48 volunters was observed and measured on MRI images.Results The height of male patella was (41.87±4.67)mm;the width was (41.08±4.30)mm,while the height of female patella was (35.34±5.15)mm,the width was (36.71±3.07)mm.The proportion of unstable Chinese patella was 37.50%.The width of male distal femur was (80.70±6.40)mm,while it was  (70.64±8.70)mm in female. Moreover,the  female femur ditch and intercondylar fossa were far smaller than male. When the ratios of width and angle for the contact area were both larger than 1,knee joints bent highly and freely.
Conclusion  The  fine fitness of patello-femoral joint,the difference of knee joint morphorlogy between men and women,and arthroscopy for knee are very important in
design of total knee prosthesis.

To discuss the effects of sevoflurane  on perioperative hemodynamics and serum cardiac troponin I(cTnI)  in patients  undergoing abdominal surgery,and offer the reference for selecting anesthetic in clinic.
Methods Forty patients with ASA Ⅱ—Ⅲ,undergoing radical gastrectomy were randomly allocated into sevoflurane group (group S) and propofol group(group P),with each 20 patients.Systemic blood pressure(SBP),diastolic blood pressure(DBP),heart reat(HR) and saturation of oxygen(SPO²) were monitored as routine.The concerntration of cTnI was examined at pre-anesthesia(T¹),post-induction(T²),beginning of operation(T³),end of operation(T⁴),the first day post-operation(T⁵) and the third day post-operation(T⁶) with radioimmunoassay.Results SBP,DBP and HR in group P at  post-induction were significantly lower than those in  group S(P<0.05).The concerntration of cTnI in group P was significantly higher than that in group S at post-induction and the first day post-operation(P<0.05).Conclusion Hemodynamics in sevoflurane group keeps steady during the induction period,and shows  lighter myocardial injury compared with propofol group,the result indicates that perioperative myocardial protection of sevoflurane is  better than that of propofol.