Objective To construct a cDNA library by using mRNA from Gloydius ussuriensis(G. ussuriensis) venom gland, to clone and analyze serine proteinase gene from the cDNA library. Methods Total RNA was isolated from venom gland of G. ussuriensis, mRNA was purified by using mRNA isolation Kit. The whole length cDNA was synthesized by means of smart cDNA synthesis strategy, and amplified by long distance PCR procedure, lately cDAN was cloned into vector pBluescript-sk. The recombinant cDNA was transformed into E.coli DH5α. The cDNA of serine proteinase gene in the venom gland of G. ussuriensis was detected and amplified using the in situ hybridization. The cDNA fragment was inserted into pGEMT vector, cloned and its nucleotide sequence was determined. Results The capacity of cDNA library of venom gland was above 2.3×10⁶. Its open reading frame was composed of 702 nucleotides and coded a protein pre-zymogen of 234 amino acids. It contained 12 cysteine residues. The sequence analysis indicated that the deduced amino acid sequence of the cDNA fragment shared high identity with the thrombin-like enzyme genes of other snakes in the GenBank. The query sequence exhibited strong amino acid sequence homology of 85% to the serine proteas of T. gramineus, thrombin-like serine proteinase Ⅰ of D. acutus and serine protease catroxase Ⅱ of C. atrox respectively. Based on the amino acid sequences of other thrombin-like enzymes, the catalytic residues and disulfide bridges of this thrombin-like enzyme were deduced as follows: catalytic residues, His⁴¹,Asp⁸⁶, Ser¹⁸⁰; and six disulfide bridges Cys⁷ -Cys¹³⁹, Cys¹²⁶ -Cys⁴², Cys⁷⁴ -Cys²³², Cys¹¹⁸ -Cys¹⁸⁶, Cys¹⁵⁰-Cys¹⁶⁵, Cys¹⁷⁶-Cys²⁰¹. Conclusion The capacity of cDNA library of venom gland is above 2.3×10⁶, overtop the level of 10⁵ capicity, The constructed cDNA library of G. ussuriensis venom gland would be helpful platform to detect new target genes and further gene manipulate. The cloned serine proteinase gene exhibits strong amino acid sequence homology of 85% to the serine proteas of T. gramineus, thrombin-like serine proteinase Ⅰ of D. acutus and serine protease catroxase Ⅱ of C. atrox respectively.

Objective To investigate the effect of pEgr-hPTEN stable transfer combined with irradiation on the proliferation and apoptosis of SHG-44 human glioma cells in vitro. Methods pEgr-hPTEN vector containing the exogenous wild type PTEN gene was transfected into SHG-44 cells under mediation of lipofectamine in vitro, positive cell clones were selected and amplified. Western blotting was used to detect the properties of PTEN expression induced by X-ray irradiation. Flow cytometry and cell growth curve were adopted to measure the effects of PTEN gene transfer combined with different doses of X-ray irradiation on cell proliferation and apoptosis of the transfected SHG-44 cells. Results Expression of PTEN protein could be enhanced by X-ray irradiation in SHG-44-hPTEN stable transfer cells. PTEN protein relative level was in dose-dependent manner within 5 Gy. pEgr-hPTEN stable transfer combined with X-ray irradiation could significantly inhibit the proliferation and induce apoptosis of SHG-44 cells. At the 8th day after irradiation with different doses of X-ray, the numbers of SHG-44-hPTEN stable transfer cells were only 30.0%-50.0% of that of SHG-44-hPTEN/0 Gy group and 7.7%-13.0% of SHG-44/0 Gy group. The percentage of early apoptotic cells of SHG-44-hPTEN group after irradiation with X-ray irradiated were 1.5-2.3 times as much as that of SHG-44-hPTEN/0 Gy group, 1.9-4.4 times as much as that of SHG-44 irradiated group and 3.4-5.1 times as much as that of SHG-44 /0 Gy group. Conclusion The apoptosis of tumor cells could be significantly enhanced and its growth could be significantly inhibited by gene -radiotherapy in vitro.

Objective To observe the curative effects of deep partial thickness derma burn injury using bone marrow mesenchymal stem cells. Methods The bone marrow stem cells were obtained from the male guinea pig and cultured in vitro. Then the mesenchymal stem cells (MSCs) with 2×10⁶ cell•mL^-1 and 2×10⁷ cell•mL^-1 were prepared, and transplanted into the wounds of the female guinea pig models with deep partial thickness derma burn injury. The wound healing area was counted and the morphological changes were observed by using HE and Masson stain. Results The wound healing in therapy groups were significantly better than that in control groups. HE stain showed that dermal and epidermal structures were clear, and cell structures of every epidermal layers were distincted and integrated in therapy groups,but cell structures of every layers in control groups were unclear. Masson stain showed that collagen was regular and new-born epidermal covered the wound in therapy groups, collagen was disorder and thicker in control groups. Bone marrow MSCs could promote the healing of wound and the effects in therapy groups were better than that in the control groups. But there was no significantly difference between MSCs with 2×10⁶ cell•mL^-1 and 2×10⁷ cell•mL^-1 in therapy groups. Conclusion The bone marrow MSCs have significant curative effects on deep partial thickness derma burn injury.

Objective To study the anti-tumor activity and the acute toxicity of (PBT) in vivo and in vitro. Methods MTT method was used to examine the inhibitory effects of PBT against several kinds of human cancer cells and the toxicity against the normal cells; H₂₂ tumor-bearing mice were used to set up animal models to measure the inhibitory effects of PBT in vivo; the conventional acute toxicity experiment of mice by gastric infusion were used to calculate the value of LD₅₀. Results The values of IC₅₀ of PBT against several kinds of human cancer cells (SMMC-7721, SGC-7901, HeLa) were 57, 209, and 193 mg•L^-1， respectively. The highest inhibitory rate (IR) of PBT against FL cells was lower than 50%. The IR in the low, middle, and high dose groups of PBT in H₂₂ tumor-bearing mice were 20.32%, 33.15%, and 52.94%, respectively. The weight of tumor in high dose group had not significant difference with that in 5-Fu group (P>0.05). The median lethal dose (LD₅₀) of PBT in mice was 1 117.38 mg•kg^-1 ,and the range of 95% confidence limit was 911.59-1 369.65 mg•kg^-1. Conclusion PBT could inhibit the growth and proliferation of human cancer cells mentioned above in vitro. Furthermore it is able to kill SMMC-7721 cell significantly. PBT could obviously inhibit the increase of tumor weight in tumor mice. PBT belongs to a kind of low toxic material.

Objective To explore the effects of mitomycin (MMC) on the expressions of P21 protein in EL-4 mouse lymphoma cells. Methods Flow cytometry and immunofluorescence staining were used to measure P21 protein expressions in changes with time and doses. Results In time-course experiments, it was demonstrated that P21 protein levels were markedly increased at 2-48 h after treatment with 2 mg•L^-1 MMC in EL-4 cells compared with control group (P<0.01, P<0.05). In dose-effect experiments, it was showed that P21 protein expressions were increased significantly 24 h after treatment with 1，2，and 4 mg•L^-1 MMC in EL-4 cells compared with control group (P<0.01). Conclusion The P21 protein expression could be increased by MMC in time- and dose-dependent manners.

Objective To investigate the neuroprotection of Venlafaxine and its mechanism. Methods The influence of Venlafaxine on the survival rates of primary cultured hippocampal neurons injured by glutamate to mimic the changes in depression was detected with MTT method；the LDH release, GSH activity, SOD activity, and MDA value were detected before and after treatment with Venlafaxine. Results Venlafaxine prevented the neurons in himppocampus from the damage induced with glutamate, elevated the neurons survival rates, reduced the LDH release, kept the cell membrane intact, elevated the SOD activity and GSH activity and decreased the MDA level. Conclusion Venlafaxine possesses the neuroprotection.

Objective To investigate the effects of intrathecal injection with U0126 on the behavior of chronic constrictive injury (CCI) rats and the c-fos expression in spinal cord dorsal horn. Methods Thermal and mechanical nociceptive thresholds were measured before CCI and then on odd-numbered days, up to and including 15th day, to assess and compare the anti-nociceptive effects of adminstration with MEK inhibitor U0126. The time course of expression of c-fos were assessed by immunohistochemical analysis. Results Intrathecal adminstration with U0126 could significantly attenuate the activation of c-fos induced by CCI and simultaneously ameliorate mechanical and thermal hyperalgesia induced by CCI. Conclusion This study shows that the activation of extracellular signal-regulate kinase(ERK) contributes to the CCI-induced neuropathic pain.

Objective To investigate the effects of Gross Saponin of Tribulus Terrestris (GSTT) on acute myocardial ischemia in rats. Methods The acute myocardial ischemia models were established by isoproternol subcutaneous injection and pituitary sublingual venous injection respectively. The effects of GSTT on the electrocardiogram T-wave of isoproternol inducing acute myocardial ischemia and the serum contents of LDH and CK of pituitary inducing acute myocardial ischemia were observed. Results GSTT with doses of 10.4,20.8,and 31.2 mg•kg^-1 could significantly improve the decreased heart rates induced by isoproternol (P<0.01, P<0.05);GSTT with dose of 31.2 mg•kg^-1 could remarkably increase the activities of serum LDH and CPK in acute myocardial ischemia rats (P<0.01).Conclusion GSTT can protect myocardium in rats with acute myocardial ischemia.

Objective To construct an eukaryotic expression plasmid containing the chimeric gene coding for the hemagglutinin-neuraminidase(HN) of newcastle disease virus(NDV) and human IL-18. Methods The enzymolysis and ligation were used to joint HN cDNA to IL-18 cDNA of pVAX1 IL-18 plasmid and at the same time the terminator of IL-18 cDNA was replaced, an eukaryotic expression vector of chimeric gene for HN and IL-18 was constructed. The recombinant plasmid was transfected into HeLa cells with liposome and the expression products were examined by hemagglutination test and Western blotting. Results The expression plasmid of pVAX1 IL-18 HN was contructed successfully confirmed by enzyme digestion identification. The expressions of IL-18 and HN chimeric genes were confirmed, and the expressed HN protein had higher hemagglutinative titer.Conclusion The constructed eukaryotic expression plasmid can express in vitro,and the expression products of the chimeric gene have good reactivity and specificity.

Objective To explore the role of 6-hydroxydopamine in the pathogenesis of Parkinson′s disease(PD). Methods The 6-hydroxydopamine(6-OHDA) was stereotaxically injected into the right striatum of rat to build the PD model. After two months, the levels of glutathione (GSH) and reactive oxygen in the striatum were measured, and the pathological changes in the substantia nigra (SN) were observed. Results The level of GSH of right side of striatum in model groups （36.85±8.64 μg•mg^-1 prot） decreased significantly and the level of reactive oxygen （58.69±9.84 U•mg^-1 prot）increased markedly. There were significant differences between the right and left striatum in animal model groups (P<0.05). Compared with the sham group and control group, there were also significant differences (P<0.05). The dopaminergic neuron number in the right SN in model group decreased significantly compared with the left SN, sham group, and control group (P<0.05). Conclusion 6-OHDA can increase the level of free radicals and decrease the level of GSH, thus leading to the oxidative lesions in SN and striatum and neuronal death.

Objective To build the model of ortholopic liver transplantation (OLT) of heart arrest donor of rat to observe the effects of HSP70 on the liver graft ischemia-reperfusion injury at different time-point after heart arrest. Methods The rats were divided into 4 groups according to the ischemic preconditioning (IP) or not, and the time of heart arrest (45 or 60 min). One hour after portal vein reperfusion,the blood samples were obtained to determine the levels of alanine amino transferase (ALT) and L-asparate amino transferase (AST), grafts samples were obtained for optical microscope observation, and the survival rate of 1 week was observed. Results The HSP70 expressions in IP groups were low at 6 h, and high at 24 h and 48 h. While there were almost no HSP70 expressions in the control group (C group) at 24 h and 48 h.At 1 h after portal vein reperfusion, the levels of ALT and AST in the IP groups (IP-45, IP-60) were significantly lower than those in C groups (C-45, C-60). At one week after the operation, in the IP groups, the structures of the hepatic lobules were very well, there were a little infiltration of lymphocytes in the portal area; while in C group there existed considerable hepatic necrosis and ballooning degeneration. In the IP groups, the survival rates at one week after operation were 62.5% and 37.5%,respectively, while in the C-45 and C-60 groups were 12.5% and 0,respectively. Conclusion HSP70 not only significantly improve liver function and graft pathological changes, but also increase survival rate of rat liver transplantation from heart arrest donors. HSP70 have remarkably protective effects on graft ischemia reperfusion injury in rat liver transplantation from heart arrest donors.

Objective To establish a multidrug resistance cell line of human hepatic carcinoma and study the mechanism of multidrug resistance. Methods Human hepatocellular carcinoma cell line BEL-7402 was induced to multidrug resistance cell line BEL-7402/ADM by intermittent administration of high dose of adriamycin (ADM). The multidrug resistance was detected by using MTT assay. The expressions of mdr-1 gene-coded P-glycoportein (P-gp), multidrug resistance-associated protein (MRP), and GSH/GST were examined by flow cytometric assay. The expressions of MDR and MRP gene were also detected using RT-PCR in both BEL-7402/ADM and BEL-7402 cell lines. Results BEL-7402/ADM was resistant to many anti-tumor agents. The IC₅₀ of ADM was 17.31 times higher than that of BEL-7402. It′s expressions of P-gp and MRP were increased significantly to (65.5±5.8)% and (34.2±3.5)%， respectively, while those of BEL-7402 were only (31.3±4.3)% and (23.4±3.1)%, respectively. There was no difference of the GSH/GST expression between BEL-7402/ADM and BEL-7402 cells. The levels of MDR and MRP mRNA expression were relatively high in BEL-7402/ADM.　 Conclusion BEL-7402/ADM is a model with multidrug resistance and the levels of MDR and MRP mRNA expressions are higher in BEL-7402/ADM than those in BEL-7402.

Objective To study the damage effect of moderate-wave ultraviolet (UVB) on fibroblast and to establish the models of cellular damage. Methods Murine fibroblast line, NIH3T3, were treated with different doses of moderate-wave UVB. MTT assay was performed to evaluate the cellular survival rate，flow cytometry was performed to detect the cell cycle，apoptosis and the production of H₂O₂. Results The cellular survival rates decreased significantly 24 h after treatment with different doses of UVB (from 0.2 to 3.0 kJ•m^-2) (P<0.05 or P<0.01)；while 1.5 kJ•m^-2 UVB induced the increase of the percentages of G₁ phase and the increase of apoptotic bodies 4,8,18, and 24 h after irridiation，simultaneously 8 h after irridation with 1.5 kJ•m^-2 UVB, the increase of H₂O₂ production was found (P<0.05). Conclusion The UVB might induce the G₁ phase arrest，the increase of apoptosis and production of H₂O₂ in fibroblast.

Objective To investigate the effects of advanced glycation end products (AGEs) and Ginkgo biloba L. on proliferation, cell cycle and transforming growth factor β (TGF-β) expression of bovine retinal pericytes, and observe the protection of Ginkgo biloba L. on pericytes. Methods MTT colorimetric assay, flow cytometry, immunofluorescence were used for analysis. Results Ginkgo biloba L. could promote proliferation of pericytes undergoing AGEs. The Ginkgo biloba L. with concentration of 125~400 mg•L^-1 could promote proliferation of pericytes obviously (P<0.01) in a dose-dependent manner. Ginkgo biloba L. could inhibit the effects of AGEs on cell cycle of pericytes and induce apoptosis. Ginkgo biloba L. decreased the number of S-phase cells, increased G₂-M phase cells, decreased the number of apoptotic pericytes obviously compared with AGEs (P<0.01). Ginkgo biloba L. also down-regulated TGF-β protein expression induced by AGEs. Conclusion AGEs can damage bovine retinal pericytes through one of the pathways of oxidative stress, and antioxidant Ginkgo biloba L. can reduce the toxic effects of AGEs.

Objective To study the effects of extraneous vascular endothelial growth factor (VEGF) on mammary carcinogenesis in resistant Copenhagen（COP）rats induced by N-nitroso-N-methylurea（NMU）. Methods VEGF were administered in a sustained releasing formula to female COP rats treated with NMU and mammary tumor incidences in VEGF-treated animals and controls were detected. RT-PCR and histopathology were used for analysis of VEGF expression and MAST cells accumulation. Results ①There was no significance difference of mammary tumor incidence between VEGF-treated animals and controls. ②Mammary epithelial cells from pubescent BUF and COP rats expressed substantial levels of VEGF messages, whereas cells prepared from 230-day old COP rats showed negligible levels of VEGF mRNA. ③Mammary epithelial cells from tumors developed in susceptible BUF rats expressed VEGF, whereas VEGF messages were barely detectable in tumors induced in COP rats. ④Enlargement of the intramammary lymph nodes with prominent MAST cells was observed in NMU-treated COP rats, but not in NMU-treated BUF rats. Conclusion These results suggest that down-regulation of VEGF expression is insufficient for resistance to mammary carcinogenesis, and negative or low expression of endogenic VEGF and enhanced immune response with MAST cells accumulation may also play a role in conferring resistance in the COP rats.

Objective To investigate the expressions of MMP-9 and MMP-9 mRNA in gastric carcinoma and their correlations with the degradation of type Ⅳ collagen. Methods The expressions of MMP-9,MMP-9 mRNA and type Ⅳ were observed by immunohistochemistry,in situ hybridization and morphometric analysis in 74 cases of gastric carcinoma.The relationships between the expressions and the clinical parameters were analysed. Results The expressions of MMP-9 and its mRNA were closely related to the invasion, differentiation and lymph nodes status of the gastric carcinoma.With the degradation of type Ⅳ collagen,the MMP-9 positive cases were increased.However,the difference among the groups had no statistical significance. Conclusion Gastric carcinoma cells have the ability to produce and secret MMP-9 which can promote the invasion and metastasis of the neoplasm. The production,secretion and function of MMP-9 were extremely controlled by the regulation of internal environment.

Objective To study the toxicity of MeHg on photoluminescent bacteria and the inhibitory effects of zinc. Methods The test of photoluminescent bacteria was used to detect the effects of MeHg and zinc on the luminescence rate of photoluminescent bacteria. Results The EC₅₀ of MeHg and zinc were 0.00248 mg•L^-1 and 0.28 mg•L^-1， respectively. With the dose increase of MeHg and zinc, the luminescence rate of the bacteria was decreased in a dose-dependent manner. An antagonism was found 15 and 30 min after administration of zinc combined with MeHg. Conclusion zinc could inhibit the luminescence rate of photoluminescent bacteria and antagonize MeHg toxicity in certain dose and time.

Objective To investigate the hemodynamic characteristics of intracranial terminal aneurysms and analysis the possible mechanism of growth and rupture of intracranial aneurysms. Methods Some images of intracranial terminal aneurysms were handled by some image processing software. The profile meshed by some computational fluid dynamics (CFD) software were used to simulate the flow and analysis wall shear stress. Results From the beginning to 0.22 T in every cardiac cycle, the wall shear stress had a change sharply to the maximum and then decreased dramatically to the zero from 0.22-0.55 T accompany with velocity. There were maximum stress and minimum stress when 0.22 T and 0.55T at neck, but meanwhile there was no wall shear stress at the wall. Conclusion Pressure is one of risk factors which will make the aneurysm rupture. CFD simulation can provide key hemodynamic information for pathophysiological mechanisms and clinical treatment of intracranial terminal aneurysms.

Objective To investigate the effects of heroin on purine nucleotides catabolism by testing the adenosine deaminase(ADA) concentrations in brain and muscle tissues in rats which were administrated with heroin. Methods The rat models of heroin administration and withdrawal were established. Concentrations of ADA in brain and muscle tissues were tested. Results When two heroin administration groups were compared with the control group, the ADA concentrations in brain and muscle tissues were increased in the 3-day heroin administration group（P>0.05）and the enhancement of the ADA concentration in the 9-day heroin administration group was more apparent（P<0.05）. When two withdrawal groups were compared with the 9-day heroin administration group, the ADA concentration in the brain wasn′t decreased apparently in the 3-day withdrawal group, while the brain ADA concentration in the 8-day withdrawal group was decreased significantly（P<0.01）. The muscle ADA concentration in the 3-day withdrawal group was already decreased （P<0.01） and the decrease of that in the 8-day withdrawal group was more significant. Conclusion The administration of heroin can enhance the catabolism of purine nucleotides in brain and muscle tissues by increasing the concentration of ADA and the effect may last for a long time in brain.

Objective To study the effects of growth hormone (GH) on morphology of cardiac muscle and skeletal muscle and hormone levels in Wistar rats. Methods The GH was given with subcutaneous injection for 15 days, the level of serum GH was determined by radiation-immune method; the body weight and the ratio of organ weight to body weight were determined; the cell appearances of cardiac muscle and skeletal muscle were observed under microscope. The control group was set up. Results The level of serum GH and rat body weight in experimental group were obviously higher than that in the control group, but the ratio of organ weight to body weight was not obviously different in two groups; musculature hypertrophy and cell nucleolus increasing were observed under microscope, there were no capillary vessel hyperplasia and inflammatory soakage. Conclusion GH can induce hypertrophy of cardiac muscle cells and skeletal muscle cells but not interstitial proliferation.

Objective To study the effects of bcr/abl antisense oligodeoxynucleotide (ASON) on K562 cell growth. Methods Eighteen nucleotides were used as target genes in bcr/abl mRNA mixed area, by different bcr/abl cDNA sequence type analysis, 18 pdy ASON fragments and 18 poly non-sense ASON fragments for special sequence control were synthesized. Incubating K562 cells with ASON or non-sense ASON fragments for a mean time, while a blank control was set up. Cytomorphology, the ³H-TdR mixed rate and the expressions of bcr/abl mRNA and P210 protein were detected. Results ASON had different inhibitory rates on cell proliferation at 3.5-30.0 μmol•L^-1. The action was smaller when ASON concentration was less than 10.0 μmol•L^-1. While the action was more significant when the concentration was larger than 10.0 μmol•L^-1. This action depends on concentration and time. But non-sense ASON and control weren′t distinct different. At the meantime, ASON made the ³H-TdR mixed rate decrease and the expression levels of bcr/abl and P210 were down-regulated. Conclusion These results show that ASON hinders bcr/abl mRNA transcription and P210 protein translation, inhibits cell proliferation and promotes apoptosis on gene levels.

Objective To investigate the effects of hydrocortisone on degradation of keratocyte induced by interleukin-1α(IL-1α). Methods The keratocyte-collagen gels were incubated with plasminogen (0.06 μg•L^-1) and different concentrations of IL-1α（0，0.1,1.0, and 10.0 μg•L^-1） or interleukin-1α (10 μg•L^-1) and different concentrations of hydrocortisone(0,1,10, and 100 mmol•L^-1) or in MEM culture medium separately 24 h later, to evaluate the collagen degradation level, and the amount of hydroxyproline(HYP) were measured. Results In the presence of plasminogen, the amount of HYP was increased with the increasing of IL-1α concentration.There were significant differences（P<0.025 or P<0.05）between 0.1,1.0,and 10.0 μg•L^-1 IL-1α groups and control group（0 μg•L^-1）,between 1.0,10.0 μg•L^-1 groups and 0.1 μg•L^-1 group,between 10.0 μg•L^-1 group and 1.0 μg•L^-1 group. In the presence of plasminogen and IL-1α(10 μg•L^-1), the amount of HYP was increased with the increasing of hydrocortisone concentration. There were significant differences（P<0.025 or P<0.05）between 1,10,100 mmol•L^-1 hydroxyproline groups and control group (0 mmol•L^-1), between 10,100 mmol•L^-1 groups and 1 mmol•L^-1 group，between 100 mmol•L^-1 group and 10 mmol•L^-1 group. Conclusion Hydrocortisone can inhibit keratocyte-collagen degradation induced by IL-1α.

Objective To explore the effects of Bismaltolatooxovanadium(BMOV) on myocardial structures in spontenously hypertensive rats while BMOV reducing the blood pressure of rats. Methods Thirsty-six spontenously hypertensive rats were randomly divided into three groups:BMOV group, nifedipine group and blank control group.They were killed after the eight weeks, their myocardial structures were observed by light microscope and electronic microscope. Results The myocardium of spontenously hypertensive rats treated with BMOV showed the follwing features: ①there was hardly collagen accumulation inside myofibrilla; ②the myofibrilla appeared normally without loss and disarrangement; ③mitochondria as well as sarcomeres showed almost normal ultrastructure. Conclusion BMOV has the protective effects on myocardium while it reduces the blood pressure in spontenously hypertensive rats.

Objective To study the expression of insulin-like growth factor-Ⅰ (IGF-Ⅰ) in focal cerebral ischemical reperfusion injury model and its contribution. Methods The focal cerebral ischemia rat models were induced using the filament method, the expressions of IGF-Ⅰ protein and IGF-Ⅰ mRNA was dectected by the method of immunohistochemical staining and in situ hybridization technique. Results The expression of IGF-Ⅰ was very low in normal tissues, while increased mainly in penumbra after cerebral ischemia. The expressions of IGF-Ⅰ protein and IGF-Ⅰ mRNA were 169.64±6.97 and 168.64±6.28,respectively, at 24 h after reperfusion and peaked to 168.86±5.39 and 170.86±5.07 at 48 h after reperfusion. Conclusion Enhanced expression of endogenous IGF-Ⅰ has protective effects on focal cerebral ischemia reperfusion injury.

Objective To investigate the effects of heroin on plasma uric acid and purine nucleotides catabolism in rat spleen by testing the activities of adenosine deaminase (ADA) and xanthine oxidase (XOD). Methods Rats were administrated with heroin by intraperitoneal injection with increasing dose to develop addiction models. Fifty male Wistar rats were divided into five groups (n=10): control, 3 d adiminigstrationg group, 9 d adiminigstrationg group, 3 d withdrawal group and 8 d withdrawal group. The uric acid concentration in plasma and the activities of ADA and XOD in rat spleen were determined. Results The concentrations of uric acid in plasma in heroin administration groups were significantly increased compared with control group (P<0.01). Compared with 9 d administration groups, the concentrations of uric acid in withdrawal groups were not decreased apparently (P>0.05). The activities of ADA in spleen in heroin administration groups were significantly increased compared with control group （P<0.05）.It was decreased apparently in 8 d withdrawal group compared with 9 d administration group. The activity of XOD in spleen in 9 d administration group was increased apparently compared with control group（P<0.05）, there were no apparently differences between withdrawal groups and 9 d administration group （P＞0.05）, but in the withdrawal groups, the XOD in spleen had taken on a decreased tendence. Conclusion Administration of heroin can enhance the activities of ADA and XOD and promote the purine nucleotides catabolism in spleen.

Objective To observe the effects of injection of ginseng fruit saponins (IGFS) on total cholesterol(TC), lipoprotein cholesterol metabolism and antioxidative activity in experimental hyperlipidemia rats. Methods The TC, lipoprotein cholesterol and lipidperoxidation (LPO) contents, prostacycline　(PGI₂) and thromboxane(TXA₂) levels, superoxidedismutase (SOD) activity and blood viscosity were measured respectively in hyperlipidemia rats which had been given IGFS 10, 20 and 40 mg•kg^-1•d^-1 ip, respectively, for fifteen days. In addition, fat accumulation in liver was observed. Results The triglyceride(TG), TC, low density lipoprotein cholesterol (LDL-c) in serum,TXA₂ in plasma, LPO in serum and liver and blood viscosity were decreased significantly, and PGI₂ in plasma and SOD in serum and liver were significantly increased after administration with IGFS (20 and 40 mg•kg^-1•d^-1) in experimental hyperlipidemia rats. Moreover, IGFS decreased ratios of TC/HDL-c and LDL-c/HDL-c, increased the ratio of PGI₂/TXA₂ and inhibit fat accumulation in liver. The content of high density lipopro tein cholesterol (HDL-c) in serum were significantly increased after adminstration IGFS （40 mg•kg^-1•d^-1） in experimental hyperlipidemia rats. Conclusion IGFS can inhibit arterioscleros by improving cholesterol and lipoprotein cholesterol metabolism, suppressing LPO and increasing antioxidation.

Objective To investigate the anti-aging effects of Seamin powders. Methods In anti-oxidation test, malondialdehyde (MDA) content , total superoxide dismutase (SOD) activity in serum from mice treated with various doses of Seamin powders were detected; in drosophila melanogaster survival test, survival time of drosophila melanogaster fed on media containing various concentrations of Seamin powders were observed and recorded, and average life span, average maximum life span and median death time were counted. At the same time, the effects of Seamin powders on body weight, temperature and survival time of swimming in cold water in yang-deficiency mice, the abilities of anti-tired and cold-resistant in spleen-hypofunction mice were observed. Results Seamin powders could significantly reduce serum MDA content in aged mice, keep body temperature and prolong the survival time of swimming in cold water in yang-deficiency mice and also extend the survival and swimming time in spleen-hypofunction mice. But it had no prominent effects on SOD activity in serum of aged mice and the average life span and average maximum life span of drosophilamelanogaster. Conclusion Seamin powders have the effects of anti-oxidation, anti-stress, but have no function of prolong the life span in drosophila melanogaster.

Objective To study the effects of muskrat and moschus of anti-inflammation and annalgesia. Methods The effects of muskrat and moschus of anti-inflammation were observed by using torsive-body method. The effects of muskrat and moschus of analgesia were observed by mice inflammation models caused by dimethylbenzene and mice models with reinforce permeability of abdominal cavity capillary caused by acetic acid. Results Both muskrat and moschus could significantly reduce the times of torsive-body in mice caused by glacial acetic acid (P<0.05 or P<0.01). And they obviously inhibited ear swelling of mice caused by dimethylbenzene (P<0.05 or P<0.01). The effects of moschus were more obvious than muskrat when they were given with the same dosage. Muskrat and moschus had obvious inhibitory effects on mice inflammation of reinforce permeability of abdominal cavity capillary caused by acetic acid (P<0.05 or P<0.01).They had no difference of effects when they were given with the same dosage（P>0.05）. Conclusion Muskrat and moschus have the same effects of anti-inflammation and analgesia.

Objective To investigate the effects of oxymatrine (OMT) on blood pressure in rats. Methods Fourty-eight Wistar rats were randomly divided into 6 groups (n=8):normal saline group,OMT groups with different doses(30,60,and 90 mg•kg^-1),propranolol+OMT 60 mg•kg^-1 group and doxazosin+OMT 60 mg•kg^-1 group. Cannula was cannulated commonly into carotid artery and connected with BL-420 BioLab System to record the blood pressure and heart rate(HR) in rats. Results Mean arterial pressure(MAP) were significantly lower than control in dose-dependent manner after intraventous adminstration of OMT(30，60，and 90 mg•kg^-1). Conclusion OMT can decrease the MAP in dose-dependent manner. OMT and doxazosin have synergetic effects on MAP.

Objective To observe the influences of ShuXinTieGao on the myocardium enzymes and inorganic elements in serum of anesthetic thoraco-opened dogs. Methods Twenty-four dogs were randomly divided into normal control group(Ⅰ),positive control group(Ⅱ),low dose of ShuXinTieGao group (0.042 g•kg^-1 ,Ⅲ), and high dose of ShuXinTieGao group (0.083 g•kg^-1,Ⅳ). The models with myocardial infarction were established through ligating the left anterior descending (LAD) coronary artery in anesthetic thoraco-opened dogs.The myocardial infarction scope　(MIS), the activitives of AST, CK, and LDH in serum and the contents of Cu,Zn,Ca, and Mg in serum were determined in model dogs. Results Compared with control group,the MIS (P<0.05,P<0.01) and the activities of serum CK, AST,and LDH (P<0.01) were obviously decreased in groups Ⅲ and Ⅳ. Compared with control group, the content of serum inorganic element Cu of acute myocardial infarction dogs was decreased (P<0.01), while the contents of serum Zn and Ca were increased　(P<0.05,P<0.01) and the content of serum Mg had no obvious change in group Ⅳ. The content of serum Zn was increased (P<0.05) compared with control group. Conclusion ShuXinTieGao can decrease the activities of the myocardium enzymes, regulate the contents of serum inorganic elements, and obviously improve the myocardial ischemia.

Objective To evaluate the protective effects of acanthopanax senticosus saponins (ASS) on mice with irradiation damage induced by X-rays. Methods The mice were randomly divided into normal control group， irradiation control group and three experimental groups. Peripheral blood WBC and PLT were detected and accounted. The transformation test of lymphocytes was done and the spleen index (SI), thymus index (TI); the activities of superaxide dismulase (SOD) and GSH-Px in serum were determined. Results Compared with the control group， TI and the activities of SOD and GSH-Px in serum in three experimental groups were increased significantly (P<0.05). Conclusion ASS have protective effects on mice with irradiation damage induced by X-rays.

Objective To evaluate the nested-reverse-transcription-polymerase chain reaction（nested-RT-PCR） for detection of cytokeratin 18 mRNA（CK18 mRNA）in diagnosis of micrometastasis in esophageal cancer and its clinical significance. Methods Nested-RT-PCR was used to detect the expressions of CK18 mRNA in the rib marrow of 50 esophageal carcinoma patients without distal metastasis(experimental group)，10 esophageal carcinoma patients with distal metastasis，10 benign esophageal disease patients. Results CK18 mRNA was not detected in benign esophageal disease patients.In the rib marrow of 50 esophageal carcinoma patients without distal metastasis and 10 esophageal carcinoma patients with distal metastasis, the positive expression rates of CK18 mRNA were 42.0%（21/50） and 100%（10/10）,respectively. The positive rate in stage Ⅳ of esophagenal cancer （100%） was significantly higher than those in stage Ⅱa（40％）, stage Ⅱb（41.7％） and stage Ⅲ（45.8%）（P＜0.05）. But there were no significant differences between stage Ⅱa, stage Ⅱb and stage Ⅲ（P＞0.05）. The positive rates in pathological types Ⅰ，Ⅱ,and Ⅲ were 41.7％,41.7％,and 42.9％, there were no significant differences between various pathological types（P＞0.05）. Conclusion In the early stage of esophageal cancer, the cancer cells have already metastasized. It is helpful to use Nested-RT-PCR to detect the expression of CK18 mRNA in the rib marrow in early diagnosis of tumor micrometastasis.

Objective To study the expressions of immune cell differentiation antigens in patients with active and inactive stages of systemic lupus erythematosus (SLE). Methods The expressions of T, B, NK, plasma and neutrophil cell differentiation antigens were studied with 18 kinds of monoclonal antibodies using indirect and direct immunofluorescence technique by cytometry from 30 normal controls and 43 SLE patients. Results The expressions of CD2, CD3, CD4, CD45RA, CD56, CD89 on immune cells were down-regulated in active stage of SLE; while the expressions of CD8, CD10, CD23, CD25, CD29, CD138 were up-regulated; other cell expressions showed no significant differences from normal control. In inactive stage the expressions of some cells reached normal except CD3, CD4, CD23, CD25, CD29, CD45RA, and CD89. The CD89 level in patients with active SLE was negatively correlated with the extent of hematuria,the correlation coefficients before and after therapy were -0.5821 and -0.7948, respectively.Conclusion The expressions of T, B, NK, plasma and neutrophil differentiation antigens are abnormal in SLE patients, especially in active stage of disease.

Objective To investigate the mRNA expressions and enzymatic activities for both matrix metalloproteinase-2（MMP-2）and tissue inhibitor of metalloproteinase-2（TIMP-2）in human gliomas and their significances. Methods The samples from 26 cases of glioma were divided into four groups according to the WHO classification, normal brain samples served as the control. The mRNA expressions for both MMP-2 and TIMP-2 were determined using reverse transcriptase-polymerase chain reaction（RT-PCR）and the enzymatic activities of MMP-2 and TIMP-2 were detected by gelatin zymography in the samples. Results In gelatin zymography, MMP-2 revealed two bands, which were localized at 62 000 (activated form) and 72 000 (latent zymogen form) of MMP-2, respectively. The latent zymogen form（72 000） of MMP-2 was not different among the four grades of gliomas. The mRNA expression level and the enzymatic activity of MMP-2 were significantly elevated with the malignant grade of the gliomas, while the mRNA expression level and the activity of TIMP-2 were markedly reduced with increasing the malignant grade of the gliomas. Conclusion The gene transcription and the enzymatic activities for both MMP-2 and TIMP-2 have changed in human gliomas with different grades of malignancy, which play an important role in the facilitation of glioma invasion.

Objective To investigate the correlations between the expression of　β-catenin and clinicopathologic parameters in bile duct carcinoma (BDC), and evaluate the potential prognostic value. Methods The expressions of β-catenin in 8 samples of normal bile ducts, 4 samples of congenital choledochal cyst and 25 samples of BDC were detected by immunohistochemistry staining　(S-P method). Results The β-catenin positive expression rate in BDC samples (100%) was obviously higher than those in samples of congenital choledochalcyst (75%) and normal bile ducts(62.5%) (Z=－3.03，P<0.01). The expressions of β-catenin in BDC were correlated with histological grade(Hc=9.35，P<0.01), infiltration grade (Hc=12.62，P<0.01)，perineural invasion (Hc=9.28，P<0.01), clinical stage(Hc=15.49，P<0.01), and were incorrelated with pathologic type(Hc=2.13，P>0.05), location(Hc=5.25，P>0.05), and severity of jaundice (Hc=6.06，P>0.05). Conclusion β-catenin is correlated with some clinicopathologic parameters of BDC and can be used as a valuable prognostic marker of BDC.

Objective To investigate the expression of inhibitor of apoptosis protein survivin and determine its role in carcinogenesis and progression in breast cancer. Methods The expressions of survivin were detected by immunohistochemistry and Western blotting in 54 samples of breast cancer (33 samples with axillary lymph node metastasis) and 15 samples of breast fibroadenma. Results The positive expression rate of survivin was 72.5% in breast carcinoma tissues, it was significantly higher than that in paracarcinoma normal tissues and breast fibroadenma tissues (P<0.05). The positive expression rate of survivin in breast cancer with axillary lymph node metastasis was obviously higher than that without axillary lymph node metastasis (P<0.05). The expression of survivin in breast carcinoma was significantly associated with histological grades, pathological stages and lymph node metastasis (P<0.05). Conclusion The expression of survivin is increased in breast cancer. The survivin is involved in carcinogenesis and progression and could be a reliable indicator to diagnose and determine the prognosis of breast carcinoma.

Objective To study the diagnostic values of S/D and △Q of umbilical artery (UmA), bilateral uterine arteries (UtA), and internal iliac artery (IlA) in maternal circulation on prediction of perinatal outcome in patients with pregnancy-induced hypertension (PIH). Methods Dopple ultrasound screening was used to detect the S/D and △Q of UmA, UtA and IlA in 48 patients with PIH and 62 cases of normal singleton pregnancy. Results The sensitivity (82.1%), likehood ratio (4.49), and Youden′s index (0.638) of UmA S/D were higher than those of UtA S/D;the sensitivtity (85.7%), specificity (82.9%), likehood ratio (5.01), and Youden′s index (0.686) of UmA △Q were all higher than those of UtA △Q; and the various indexes of UmA △Q and UtA △Q were higher than those of UmA S/D and UtA S/D; each index of IlA S/D and IlA △Q was lower. Conclusion The diagnostic value of △Q for perinatal outcome in patients with PIH is superior to S/D; the diagnostic values of IlA S/D and IlA △Q are lower than those of UmA and UtA.

Objective To study the effects of Shengmai injection on alleviating myocardial ischemia and preventing the reperfusion injury when it was applied as the additive of the perfusion liquid in the process of cardiac surgery with cardiopulmonary bypass (CPB). Methods Sixty cardioetomy patients were randomly divided into experimental group (n=30) and control group (n=30). In the experimental group, the Shengmai injection was added to the reformative Thomas solution.The serum CK-MB and SOD levels were determined in experimental and control groups.The changes of myocardial ultrastuctures were observed under electron microscope. Results The level of CK-MB was significantly lower in the experimental group than that in the control group(P<0.05 ).The level of SOD in experimental group was significantly higher than that in the control group(P<0.05).The result of mitochondria morphological measurement indicated that the mitochondrial face number density and specific surface were significantly higher in the experimental group than those in the control group (P<0.05 or P<0.01). Conclusion Shengmai injection acted as the additive of the the cold stopping-pulse solution could protect the myocardial membrane and the ultrastructure of myocardium and debate the damage in the cardioetomy and eliminate the oxygen free radical.

Objective To detect the association between the polymorphism of tumor necrosis factor promoter gene (TNF-308) and asthma. Methods The technique of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was used to examine the allele frequencies of TNF-308 in 50 asthma patients and 80 normal controls. Results The allele frequency of TNF-308 gene was not significantly different between asthma group and normal control group （χ²=0.047, P>0.05）. Meanwhile, the frequencies of three genotypes （GG,GA and AA）of TNF-308 gene were not significantly different in asthma group (45/50, 5/50 and 0/50) and normal control group (71/80, 9/80 and 0/90)（χ²=0.049,P>0.05）. Conclusion There is not association between the polymorphism of tumor necrosis factor promoter gene (TNF-308) and asthma.

Objective To discuss the effects of kinesitherapy on osteoporosis in different age females. Methods The calcaneus ultrasonic conduct velocities in all subjects were determined by CM-100 Ultrasonic Calcaneus′s Density Mensurating Apparatus, and the interfix matrix analysis of body characteristis and bone densit (SOS value) was performed. Results The effects of sports on bone densities had significant difference between sports group (group 1) and non-sports group (group 2) in 20,40 and 50 years subjects (P<0.01 or P<0.05); the grip and SOS value had significant differences between group 1 and group 2 (P<0.01); the two groups all had negative correlation with the age (group 1: r=-0.781, P<0.01; group 2: r=-0.59,P<0.05), the two groups all had positive correlation with the age and BMI (group 1:r=0.411,P<0.01; group 2: r=0.28,P<0.05),but negative correlation with age and grip (group 1: r=-0.285,P<0.05; group 2:r=-0.326, P<0.01). In the group 2, SOS had positive correlation with　Weight and GS (r=0.225,P<0.05; r=0.308, P<0.05). Conclusion Sports have definite advanced effects on definite age females. Body characteristics have definite correlation with bone density.