Objective To construct pSUPER-SVV vector and obtain its expression in HeLa cells, in order to interfere the expression of survivin. Methods Oligonucleotide sequences specific for survivin were designed for RNA interference (RNAi), after annealed they were ligated to pSUPERbasic vector. After sequencing, the vector of pSUPER-SVV was transfected into HeLa cells by LipofectAmine. The expression of survivin was detected by RT-PCR and FCM. Results The RNAi expression vector of survivin was constructed successfully, which depressed the expression of survivin efficiently. Survivin was significantly depressed on the level of mRNA in HeLa cells compared with the control groups (P<0.05), and the expression of survivin was down-regulated to 31.62%, there was significant difference compared with control group (P<0.001). Conclusion The RNAi vector targeted to survivin gene　is successfully constructed, survivin is depressed on the levels of mRNA and protein in HeLa cells transfected by the vector.

Objective To establish a prokaryotic expression system for expressing mouse enterokinase which could recognize and cut the sequence of (Asp)₄-Lys in order to provide a technique platform for application of recombinant enterokinase. Methods The cDNA coding sequencing of enterokinase light chain was amplified by RT-PCR from mouse dodecadactylon mucosa and cloned into pET32a expression plasmid. The recombinant enterokinase light chain was expressed in BL21(DE3) and purified with Ni-affinity chromatography.Results The cDNA sequence amplified was identical with that in GenBank and possessed more than 75% homologous in amino acid sequence with that from human and cow. The recombinant mouse enterokinase light chain could be well expressed in Ecoli, but most of them were inclusion. Almost all recombinant enterokina se proteins were polymers after purification by Ni-affinity chromatography. Conclusion Recombinant mouse enterokinase can be expressed in Ecoli in inclusion manner so that the prokaryotic expression system for recombinant enterokinase with native conformation needs to be optimized.

Objective To establish the mouse nonspecific keratitis model by corneal suture combined with subconjunctival injection in order to analyze the relationship between B7-1 mRNA expression and corneal inflammation and discuss the inhibition of nuclear factor-kB（NF-kB）inhibitor on B7-1 mRNA expression. Methods The corneal suture or combined with subconjunctival injection models were constructed in BALB/C mice.The corneas of each group were excised at the 1st，2nd，7th，14th day postoperatively and the B7-1 mRNA expressions were investigated by reverse transcription polymerase chain reaction (RT-PCR). Results At 1st, 3rd and 7th day postoperatively the expression of corneal B7-1 mRNA increased by the inducement of corneal suture combined with lipopolysaccharide (LPS) subconjunctival injection than that by the corneal suture (P<0.05). At 14th day postoperatively there was no significant diffe rence of the expression of corneal B7-1 mRNA between two groups (P>0.05); At 1st，7th，14th day postoperatively the expression of corneal B7-1 mRNA decreased, in the group of inducement of corneal suture combined with LPS and pyrrolidine dithiocarbamate (PDTC) subconjunctival injection compared with the group of inducement of corneal suture combined with LPS subconjunctival injection (P<0.05), but it increased at 3rd day postoperatively. Conclusion A high expression of B7-1 mRNA can be induced by the corneal stuture combined with LPS subconjunctival injection, which has a close relation with the deteriorated inflammation. The inhibitor (PDTC) of NF-kB can inhibit the expression of B7-1 mRNA. The activation of NF-kB has a great influence in the regulation of B7-1 mRNA expression. 

Objective To investigate the toxicities and inhibitory effects of three kinds of kegin-type novel heteropoly compounds which were synthesised by molecular assembly interactions on influnenza A virus(H1N1) in vitro. Methods The toxicities and anti-influnenza virus activities were tested by methods of CPE-MTT and chick embryos culture. Results TC₅₀ in MDCK cells and LD₅₀ in chick embryos of three kinds of kegin-type novel heteropoly compounds were higher than those of amantadine. TI in MDCK cells and chick embryos were both higher than that of amantadine. Conclusion The three kinds of kegin-type novel heteropoly compounds have lower toxicities and higher anti-virus effects on influnenza A virus(H1N1) than amantadine.Through chemic modifications, the toxicity of amantadine can be decreased and the activity of amantadine can be increased.

Objective To investigate the protective effects of oral aprotinin on acute hepatic injury in rats. Methods Sixty Wistar rats were divided into 6 groups:normal control group (Ⅰ) , model control group (Ⅱ), low,middle, high doses of aprotinin groups (Ⅲ，Ⅵ，Ⅴ) and diammonium glycyrrhizinate control gro up (Ⅵ). The changes of activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), glutathione (GSH) and the liver weight coefficent were measured.The pathologic changes of liver tissue were measured. Results The serum ALT, AST, NO concentrations in rats and the liver weight coefficient in Group Ⅱ were increased significantly compared with Group Ⅰ (P<0.001, P<0.001, P<0.01, P<0.001), the serum GSH concentration in rats in Group Ⅱ was decreased significantly compared with Group Ⅰ (P<001). The serum ALT, AST, NO concentrations in rats in Group Ⅲ , Group Ⅳ and Group Ⅴ were decreased significantly compared with Group Ⅱ (P<0.001, P<0.001,P<0.05). The serum GSH concentrations in rats in Group Ⅲ , Group Ⅳ and Group Ⅴ were increased significantly compared with Group Ⅱ (P<0.01). The liver weight coefficients in Group Ⅲ , Group Ⅳ and Group Ⅴ were decreased significantly compared with Group Ⅱ(P<0.01,P<0.05,P<0.001). The order of pathologic change levels of liver tissue of acute hepatic injury was Group Ⅴ，Group Ⅳ，Group Ⅲ， Group Ⅱ. Conclusion The oral aprotinin has significantly protective fun ction on rats with acute liver injury.

Objective To explore the influence of different domains of STAT6 expressed by the vector pGEX-6p-2 on signal transduction. Methods Different domains of STAT6 were amplified by PCR technique and ligated respectively into the prokaryotic expression vector. Recombinant DNA was transformed into HB101 cells. Transformants were lysed by super sonic method and lysates run on SDS-PAGE. Results Western blotting analysis showed a high level expression (60 000, 56 000 and 50 000) of different domains of STAT6 SHST,DST and NST, and the empty vector only had GST expression.Conclusion Expression vector pGEX-6p-2 can be used for high l evel expression of different domains of STAT6.

Objective To observe the effect of enzymes-hydrolyzed wheat bran (EHWB) on antioxidant system in diabetic rats.Methods Diabetes mellitus models were induced by alloxan inject ion and EHWB was used to test its effect on anti-oxidative capacity; the EHWB was given intragastrically (i.g.), sodium ferulate (SF) as a positive group. The level of blood glucose, total anti-oxidative capacity (T-AOC), the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and xanthine oxidase (XOD), and the content of malondiadehyde (MDA) were detected respectivel y in serum and liver of rats. Results EHWB significantly decreased the level of blood glucose and increased the level of T-AOC, as well as the activities of GSH-Px and SOD , and decreased the activity of XOD and the content of MDA. However, there was no difference between the groups feeding SF and EHWB. Conclusion EHWB can be used to decrease blood glucose and improve antioxidant ability in diabetes mellitus rats，the effect of EHWB is similar to SF. The effect of decreasing blood glucose of EHWB is related to protecting liver.

Objective To observe the effet of gross saponin of tribulus terrestris（GSTT） on hemodynamics and oxygen metabolism in anesthetic thoraco-opened dogs. Methods Thirty dogs were randomly divided into negative control, positive control, and experimental groups(GSTT of 6.26,12.52, and 25.00 mg·kg^-1)（n=6）. The administration of intravenous injection was used. The effects of GSTT on the indexes of the hemodynamics and oxygen metabolism in anesthetic thoraco-opened dogs were observed. Results Compared with controls GSTT increased myocardial blood flow (CBF)(P<0.01), decreased mean arterial blood pressure (MBA), cornary resistance(CR) and total peripheral vascular resistance (TPVR)(P<0.05 or P<0.01), reduced the content and index of oxygen consumption of myocardium (P<0.01) and myocardial oxygen uptake rate(P<0.01). Conclusion GSTT can significantly accommodate and ameliorate the heart function of anesthetic thoraco-opened dogs.

Objective To observe the effect of panax quinquefolium fruit sa ponin (PQFS) on hemodynamics and correlative indexes in the coronary artery liga ted dogs. Methods The dogs were divided into five groups randomly (n= 6): control group,gingkgo leaf tablet group (24 mg·kg^-1), 24,12 and 6 mg ·kg^-1 PQFS groups. The myocardial infarction size (MIS )and the changes o f the serum enzymes were determined by using the acute myocardial infarction mo dels with the ligation of left anterior descending coronary artery of dogs. The parameters of hemodynamics and myocardial oxygen metabolism were measured in the anesthetic dogs with thoracotomy. Results Compared with control group, the myocardial blood f low was increased（P<005）,the coronary resistance and myocardial oxygen w ere decreased（P<005）in dogs treated with PQFS of 24, 12 and 6 mg·kg^-1. The MIS and the activities of serum glutamic oxaloacetict ransaminase enzy me　(AST), creatine kinse (CK) and lactic dehydrogenase (LDH-L) were decreased （P<001,P<005）in acute myocardial infarction dogs treated with PQFS of 24,12 and 6 mg·kg^-1. The activities of superoxide dismutase (SOD) wa s increased（P<001,P<005）, but the content of serum malonyl dialdeh yde (MDA) was decreased（P<001,P<005）. Conclusion PQFS has protective effect on myocardial ischemi a through decreasing myocardial oxygen consumption and increasing myocardial blo od flow in dogs.

Objective To examine the effects of Cyclovirobuxinum D sustained-release tablet on the serum enzymes and trace elements and myocardial morphology in acute myocardial infarction dogs. Methods Thirty heterogeneous dogs were divided into five groups (n=6):model group, control group with curative drug,the Cyclovirobuxinum D sustained-release tablet groups with 0.16,0.32 and 0.64 mg·kg^-1, respectively. The changes in the activities of the enzymes and the contents of serum trace elements （Cu²⁺,Zn²⁺,Mg²⁺）were determined in the myocardial infarction models by ligating the left anterior descending branch (LVD) of dogs. The morphological changes of myocardial cells was observed by using HE staining. Results Compared with model group, the activities of AST, CPK, LDH and the content of Cu²⁺ were decreased significantly in the groups with different doses of Cyclovirobuxinum D sustained-release tablet （P<0.05,P<0.01）, whereas the contents of Zn²⁺ and Mg²⁺ were incre ased（P<0.05,P<0.01）. Consequently, Cyclovirobuxinum D sustained-release tablet ameliorated the pathomorphological injury of myocardial cells. Conclusion Cyclovirobuxinum D sustained-release tablet has apparently protective effects on myocardium in canine experimental myocardial infarction models.

Objective To observe the pathological changes of hippocampus and myelin in rats with frontal cereberal ischemia and reperfusion after treated with methylcobalamine and to discuss the mechanism of the cerebral protective eff ect of methylcobalamine.Methods Cerebral ischemia and reperfusion models were made by clamping bilateral carotid arteries. Forty-five male Wistar rats were randomly allocated to sham-operation group,ischemia- reperfusion group and methylcobal amine group(n=15). 1,2,4 weeks after reperfusion the rats in each group were divided into three groups(n=5). The pathological changes of hippocampus and myelin were observed by using light microscope, HE staining, LFB staining and electron microscope. Results The pyramidal neurons in hippocampus in ischemia and reperfusion groups expressed putrescence and myelin necrosis or demyelination at 1st week,and the histomorphological changes at 4th week were significant than that at 2nd week.The followings were observed in electron microscope: dimness, partly solubilization and collapsablity in myelin. The pyramidal neurons in hippocampus of therapy groups were similar to normal one and myelin expressed compaction and shipshape. The changes were more obvious at 4th week. Conclusion Methylcobalamine can obviously decrease the ischemia lesion of hippocampus and myelin and protect the brain function.

Objective To study the protective effects of total glycosides of Pacony (TGP) on acute myocardial infarction in dogs. Methods Thirty domestic dogs were randomly divided into 5 groups (6 in each group ): model control group, positive control group(ISM), and TGP groups with doses of 2.0, 4.0, and 8.0 mg·kg^-1, respectively. The descending branch of left coronary artery was ligated to construct the model of myocardial infarction in the anesthetic thoraco-opened dogs. The effect of TGP on epicardium electrocardiogram (EECG) , myocardial infarction size (MIS) and serum myocardial enzymes were observed. Results Compared with controls, the ischemia degree (∑－ST，P<0.05，P<0.01), ischemia area(N-ST，P<005) and infarction area were reduced in TGP groups with various doses, the activities of serum creatinephosphoric kinase (CPK) and lactate dahydrogenase (LDH) were deceased (P<0.05，P<0.01). Moreover，the levels of free fatty acids (FFA) and lipoperox dation (LPO) were decreased (pcaos,pcaol), the activities of serum superoxide dismutase (SOD) and glytathion-peroxdation (GSH-Px )were increased (P<0.05，P<0.01). Conclusion TGP has effects on protection of the ischemic myocardial tissue.

Objective To study the antitumor immune activities of the Lewis lung cancer cells modified by interleukin-12 (IL-12) gene in vivo.Methods The Lewis lung cancer cells were transfected with AdCMIL-12 by differe nt multiplicity of infection (MOI). C57BL/6 mice were treated by subcutaneous vaccination with the differently disposed Lewis lung cancer cells,and then challenged by Lewis lung cancer cells and B16 melanoma， respectively. The tumor growth was observed. Results The tumor growth in the vaccine group was inhibited significantly, not only when C57BL/6 mice were treated with the differently disposed Lewis lung cancer cells, but also when C57BL/6 mice were challenged again by Lewis lung cancer cells after subcutaneous vaccination for 6 d. But the tumors occurred when C57BL/6 mice were challenged again by B16 melanoma. These results indicated that the tumor vaccine had the tumor specificity. Conclusion Immunization with the Lewis lung cancer cells transfected by AdCMIL-12 could induce potent antitumor immune activity specifically, and may have significant therapeutic effects on the recurrence and the metastasis of lung cancer.

Objective To investigate the effect of As₂O₃on normal mouse bone marrow stromal cells(BMSCs). Methods BMSCs and colony-forming unit-granulocyte macrophage(CFU-GM) were cultured in vitro to observe the quantitative changes of colony-forming unit-fibroblast (CFU-F) and CFU-GM after BMSCs were treated with different concentrations of As₂O₃respectively;the quantitative changes of CFU-GM supported by BMSCs and granulocyte colony stimulating factor(G-CSF) and IL-11 secreted by BMSCs were also observed after the progenitor cells supported by BMSCs were treated with As₂O₃ for 3，5 and 7 d，respectively. Results The productions of CFU-F and CFU-GM supported by BMSCs were decreased obviously when treated with 7 μmol·L^-1 As₂O₃, also the levels of G-CSF and IL-11 secreted by BMSC(P<0.05). The levels of G-CSF and IL-11 secreted by BMSCs were increased obviously if treated with 3 μmol·L^-1 As₂O₃ continuously for 3 or 5 d, whereas without increasing obv iously when treated with 3 μmol·L^-1 As₂O₃ continuously for 7 d and later (P<0.05). Conclusion Lower concentration of As₂O₃has not significant inhibitory effects on clone forming of BMSCs and the productions of CFU-GM supported by BMSCs, but stimulates BMSCs to secret G-CSF and IL-11. The concentration of As₂O₃ causing the max level of G-CSF and IL-11 is 3 μmol·L ^-1.The administration time when achieved the max levels of G-CSF and IL-11 is 5 d. The higher concentration of As₂O₃and the elongation of the administration time can decrease the secretion of G-CSF and IL-11.

Objective To study the direct inhibition of Ginsenoside-Rg3(GS-Rg3) on human keloid fibroblasts cultivated in vitro and its possible mechanism. Methods The effects of GS-Rg3 with different concentrations (25,50,100 and 200 mg·L^-1) on proliferation activity of fibroblast hyperpla sia at different time were detected by viable count and MTT chromometry. Results Compared with control group, GS-Rg3 inhibited the proliferation of keloid fibroblasts. The inhibition increased with the prolongation of time and the increasing of concentration. And the max inhibitory concentration was 100 mg·L^-1. Conclusion Gs-Rg3 can inhibit the proliferation of human keloid fibroblasts cultivated in vitro. And the inhibition will increase with the prolongation of time and the increasing of concentration.

Objective To observe the effects of lysophosphatidic acid (LPA) on arrhythmia in acute myocardial infarction (AMI) rats. Methods AMI models were prepared by ligating the left coronary branch of rats. 60 Wistar rats were divided into 5 roups:sham-operated (SO), AMI, AMI+LPA,AMI+pertussis toxin (PTX) and AMI+PTX+LPA. ECG were recorded, HR and VPBs were observed as important indexes. Results Incidence of tachycardia and VPBs of unit time in AMI group were increased compared with sham-operated group (P<0.01).Incidence of tachycardia and VPBs of unit time in AMI+LPA group were increased compared with AMI group (P<0.05). Incidence of tachycardia and VPBs of unit time in AMI+PTX group were decreased compared with AMI+LPA group,it showed the effect was blocked by PTX, a G-protein-coupled receptor blocking agent. Conclusion LPA can promote the occurrence of arrhythmia in AMI rats.

Objective To study the effects of zinc and chromium on declining blood glucose level and improving antioxidant activity in diabetic mice. Methods The models of experimental diabetic mice were established by abdominal cavity injection of alloxan with the dose of 200 mg·kg^-1 . The mice were divided into 4 groups: zinc, chromium, normal control and experimental control group. The mice in zinc and chromium groups were respectively administered orally with ZnSO₄ and CrCl3 solution with the dose of 5.1 mg·kg ^-1·d^-1(Zn²⁺) and 40 μg·kg^-1·d^-1(Cr³⁺). Xanthine-oxidasemeasurement method was used to measure the quantity of SOD in hepatic tissues. Colorimetric method was used to measure the activities of GSH-Px and T-AOC in hepatic tissues. Thio-barbituric acid was used to measure the level of MDA. After four weeks, blood glucose, T-AOC, MDA, SOD, GSH-PX were detected. Results Compared with experimental group, blood glucose levels in zinc and chromium groups were decreased（P<0.01）, the activities of SOD and GSH-Px as well as T-AOC were increased（P<0.05 or P<0.01）. Meanwhile, the level of MDA was declined remarkably（P<01）. Compared with normal control group, the activities of SOD, GSH-Px and T-AOC were decreased (P<0.01）. Conclusion Zinc and chromium can decrease the blood glucose and improve the antioxidant activity in diabetic mice.

Objective To discuss the effects and mechanism of Rutongning capsule on hyperplasia of mammary glands in rats. Methods 50 female Wistar rats without pregnancy were randomly divided into normal group，model group，low dosage group，middle dosage group and high dosage group（n=10）. Estradiol and progesterone were used to induce hyperplasia models of mammary glands in rats. The rats with hyperplasia of mammary glands were administered by gavage with Rutongning. The diameter of the second twin nipples of the rats was measured and the structure of the mammary gland tissue was observed. The level of hormone in blood was detected by radioimmunity and posit ive cells of estrogen receptors（ER） and progesterone receptors(PR) were determ ined by immunohistochemical technique. Results Compared with model group, estradiol, prolactin, follicle stimulating hormone in middle and high dosage groups were decreased (P<0.05),and luteinizing hormone , progesterone were increased (P<0.05) ,and the diameter of the second twin nipples of the rats was reduced (P<0.05). Rutongning made a cini atrophy and make gland ducts, and their volumes became small significantly (P<0.01). Rutongning decreased the number of ER and PR positive cells significantly. Conclusion Rutongning could rectify the secretion of hormone in rats and weaken the biological effects of estrogen and progesterone on mammary glands，so the hyperplasia mammary glands could get recovery.

Objective To study the effects of Bis(maltolato) oxovanadium (BMOV) on renal structure of spontaneously hypertensive rats(SHRs) and its mechanism. Methods Thirty-six SHRs were divided into three groups at random: blank group, Nifedipine group and BMOV group. The light microscope and electron microscope were used to examine the pathologic changes of the kidney in rats. Results The rat glomerular mesangial expansion was found and the lume n of glomeruler capillary nearly closed in bland group, while in BMOV group the pathological changes mentioned above were unobvious. Conclusion BMOV may have protective effects on kidney in SHRs.

Objective To investigate the effect of selenium (Se) and vitamin E (VE) on collagen Ⅰ synthesis in cultured cardiac fibroblasts (CFbs) treated with norepinephrine (NE). Methods CFbs were obtained by the method of enzymatic digestion culture. Cells and culture medium were collected after 48　h，then the hydroxyproline (HYP) and collagen Ⅰ content as well as the expression of α1 ［Ⅰ］ col1agen mRNA were examined. Results The contents of HYP in Se, VE and Se+VE groups were (64.88±1.25)，(47.60±1.59) and (41.34±1.73) mg·L^-1 prot，respectively，all of them were lower than that in NE group（P<005）, which was ( 68.27±1.89) mg·L^-1 prot.The contents of collagen Ⅰ in Se, VE and Se+V E groups were (0.67±0.05)，(0.40±0.03)，(0.39±0.04), respectively; they were lower than that in NE group (0.90±0.15)（P<0.05）. Conclusion Se and VE can inhibit the synthesis, secretion of HYP and type Ⅰ collagen，and they can play in synergetic effect when they are used together.

Objective To study the effects of exogenous phosphatidic acid (PA) on calcium current of ventricular myocytes.Methods The Langendorff perfusion method was used to isolate the ventricular myocytes of guinea pigs and the ventricular myocytes of guinea pigs were randomly divided into control group, three dosage groups of PA 0.01, 0.1 and 1.0 μmol ·L^-1and group of PTX 0.1 mg ·L^-1 plus PA 1.0 μmol·L^-1and group of H-7 2 μmol·L^-1plus PA 1.0 μmol·L^-1. The whole cell voltage clamp technique was used to record the calcium current of isolated ventricular myocytes of guinea pigs. Results 0.01, 0.1 and 1.0 μmol·L^-1 PA enhanced the calcium current from （288.70±28.33）pA,（290.83±25.12）pA and（279.54±31.22）pA before administration to（304.10±28.31）pA（P>0.05）,（349.80 ±30.13）pA (P<0.01) and（388.10±28.12）pA (P<0.001) after administration. The effects of PA on the calcium current was blocked by PTX (a G protein-coupled receptor blocking agent) and H-7 (a PKC blocking agent). Conclusion PA can promote the opening of calcium channels of the ventricular myocytes by the pathway of membrane receptor-G protein-PKC in a dose-dependent manner.

Objective To study the effects of transforming growth factor-β1 (TGF-β1) on mesangial cells growth and expression of plasminogen activator inhibitor-1(PAI-1) in cultured rat mesangial cells stimulated by different levels of human recombinant TGF-β1. Methods The proliferation of mesangial cells was determined with MTT assay and FACS analysis. The levels of PAI-1 protein and mRNA were analyzed with Western blotting analysis and RT-PCR respectively. Results Compared with control group, 0.1 and 0.5 μg·L^-1 TGF-β1 stimulated mesangial cells proliferation, the proliferation rate of mesangial cell was (146.6±10.2)% and (134.6±14.5)% respectively, and the expressions of PAI-1 mRNA and protein also increased (P<0.05). The proliferation rate was （75.2±13.2%） and （67.5±11.1）% in the group treated with 5 and 25 μg·L^-1 TGF-β1, the expressions of PAI-1 mRNA and protein were constant. Conclusion Low level of TGF-β1 can trigger the proliferation of rat mesangial cells, induce the expression of PAI-1, and high level of TGF-β1 can inhibit the proliferation of mesangial cells.

Objective To study the effects of ginseng fruit saponins（GFS）on the coronary circulation and cardial oxygen metabolism in dogs with acute myocardial infarction. Methods The acute myocardial infarction model was induced by ligating the left anterior descending coronary of dogs for 6 h. 90 min after treatment with GFS, the blood flow of coronary artery was determined by using electromagnetism flowmeter with MP-27 type. At the same time，the blood oxygen co ntents of artery and vein were determined by using analysis instrument of blood gas with CORNIHG 178 type. Results After treated by GFS (in a dosage of 5 and 10 mg·kg^-1 iv drip for 90 min)，the myocardial blood flow (MBF) was increased and coronary vascular resistance (CVR) was decreased significantly. Meanwhile the cardiac oxygen consumption and myocardial oxygen utilization rate were also decreased, but cardiac oxyg en consumption index had not changed significantly. Conclusion GFS has protective effects on acute myocardial ischemia through improving the coronary circulation, increasing myocardial blood supply and decreasing cardiac oxygen consumption and myocardial oxygen utilization rate. 

Objective To explore the pathogenesis of type 1 diabetes mellitus (T1DM) induced by different doses of streptozotocin (STZ) and its relations with autoimmune response. Methods Forty KM mice were divided into control group (group Ⅰ) and 20,40,80 mg·kg^-1 STZ treated groups(group Ⅱ,Ⅲ,Ⅳ). Animal models of diabetes mellitus were induced by different doses of STZ through intra peritoneal injection. Blood and urine glucose (Glu) levels were detected with glucose detecting kit and urine analyzing papers respectively. Pancreatic histological changes of islets were observed under light microscope. The level of insulin autoantibodies (IAA) in serum was detected by indirect ELISA. The functions of Tlymphocytes in mouse spleen and thymus were detected by Lymphocytes Proliferation Assay (MTT method). Results The blood glucose in group Ⅰ changed nothing. The blood Glu levels in group Ⅱ and Ⅲ increased as times went by. Four weeks after injection of STZ, the urine Glu in group Ⅰ were all negative, in group Ⅱ 5 negative and others >+, in group Ⅲ 2 negative and others >++, in group Ⅳ all >+++. The changes and degrees of pancreatic islet β cells injured by difference doses of STZ were different. The IAA levels in groupⅡand Ⅲ were higher than that in control group　(P<0.05), but there was no difference between group Ⅳ and control group. Spleenocytes (mature T lymphocytes) were highly proliferated by ConA and the thymocytes (immature T lymphocytes) were lowly proliferated by Con A in groupⅡand Ⅲ. However, there was no difference between group Ⅳ and control group. Conclusion Three different doses of STZ (20,40,80 mg·kg^-1) can induce different kinds of DM. High dose 80 mg·kg^-1) STZ injection can induce T2DM in mice, and low dose (20,40 mg·kg^-1) and multiple injection can induce T1DM. The better dose is 40 mg·kg^-1.

Objective To investigate the changes of nerve-action in mice under methylmercury (MMC) exposure in one unit at fetal period and at adult period. Methods 44 mice (C57BL/6Cr) were divided into four groups:control group,exposure group at fetal period (prenatal group ),exposure group at adult period (adult group),and expsoure group at fetal period and adult period (repeative exposure group). Every group included 11 mice. The mice in prenatal group were given 3 mg·kg^-1 MMC for 3 d during days 12-14 gestation.The offsprings were given 12 mg·kg^-1 MMC when they when they were 13 weeks old in adult group. The mice were given MMC both at fetal period and adult period in repeative exposure group, while the same dose of physiological salt solution was given in control group. 10 d after administration of MMC at adult period, Open-field and Morris water-maze nerve action and brain path-histological test were performed. Results In Open-field test, the inportant index moving distance of appraise nerve action had a marked decrease in prenatal group, however,there was a increase in adult group, there was a significant difference between two groups （P<005）; there was no nerve action change in repeative exposure group. In Morris water-maze test, there was significant difference between prenatal group and adult group（P<005）. The repeative exposure group had a similar result with the prenatal group. The influence of exposure after growing had no meanings in statistics. The pathological results indicated that the mitochond ria and globus palidus of pallium had lots of denaturalize neurocytes. Conclusion Repeative methylmercury exposure in both fetal and grown-up period mice does not aggravate the influence on nerve-action; the damage on the brain of fetal mice may have compensative influence if it is damaged again after grow up.

Objective To study the effects of methylmercury chloride (MMC) on protein kinase C （PKC） activities in rat developing cerebellum. Methods The animals in experimental groups were fed standard ra t chow with 0.75 mg MMC (ExpⅠ),1.5 mg MMC (ExpⅡ) and 3.0 mg MMC (ExpⅢ) respectively for 90 d before gestation to 30 d post parturition. Cerebella of pups from each group on postnatal days (PND) 3, 7,17,21 and 30 were dissected. All the samples were separated into cytosol and membrane subcellular fractions and assay ed for PKC activity by the improved method from Takai′s. Results Membrane and cytosolic PKC activities of pups′ cerebella from certain experimental groups were significantly higher than those of corresponding control group, PKC activities of rats from ExpⅡ, ExpⅢ and PND 3,7,14 in ExpⅠwere significantly higher than those in control group (P<0.05).Conclusion The damage of the rat developing cerebellum by chronic exposure to MMC is possibly correlative with the enhanced effect of MMC on PKC activity.

Objective To study the antagonsim of Chinese medicine herbals on the chromsome breakage induced by Bleomycin (BLM)，which was composed of ginseng，cortex acanthopanasia radices (CAR)，pachyna cocos Frand angelica at certain concentration. Methods The mutagen sensitivety test was used to detect the eff ect of BLM on chromatid breaks per cell from peripheral blood lymphocytes of 20 healthy subjects. And the infuence of Chinese medical herbals on the stability of chromsome was studied. 50 cells per sample were observed under microscope and the n the average b/c value (chromatid breaks per cell) of every group was calculated.The all b/c values were analyzed by statistical method. Results There was obvious difference of the b/c value between BLM group and negative group (P<0.01). Chinese medicine herbals showed obviously inhibitory effect on the inducement of BLM (P<0.01). Conclusion Chinese medicine herbals can enhance the ability to anti-carcinoma of people， which acts as stable substance to chromosome. So it can prevent the occurrence of tumor.

Objective To examine the genetic toxicity of Agrimonia Pilosa Ledeb on male mouse reproductive cells.Methods Sperm abnormality test, sister chromatid exchange (SCE) test in mouse spermatogonia and unscheduled DNA synthesis (UDS) test in mouse sperms were used.Thirty mice were divided into six groups (5 mice per group): negative control（NS）, Mitomycin C (MMC) or methyl methyl sulfonate (MMS) group，and Agrimonia Pilosa Ledeb groups(1.0,2.0,4.0 and 8.0 g·kg^-1). Results Sperm abnormality or SCE had no significant differences between test groups and control group (P>0.05),indicating that Agrimonia Pilosa Ledeb could not induce any UDS in mouse sperms. Conclusion Agrimonia Pilosa Ledeb has no genetic toxicity in male mouse reproductive cells at the range of doses that are applied under the experimental conditions.

Objective To investigate the genetic association between polymorphism of PLA2G4A gene in chromosome 1 and schizophrenia in the Han people of Northern China. Methods The method of PCR-RFLP was conducted to examine the genotypes of PLA2G4A gene in samples of 235 Chinese Han family trios consisting of fathers, mothers and affected offsprings with schizophrenia. Then the analysis of haplotype relative risk (HRR) and transmission disequilibrium test (TDT) were used to deal with the data statistically. Results The genotypic frequency of PLA2G4A gene did not deviate from Hardy-Weinberg equilibrium in both case and control groups（P>0.05）; HRR analysis did not show a significant difference in the allelic (A and G) frequency of PLA2G4A gene between transmit and untransmit (P＞0.05); TDT analysis suggested that the possibility of the two different alleles transmitted from the heterozygotic parents didn′t deviate from 50%（P＞0.05). Conclusion The PLA2G4A gene in chromosome 1 may be not the susceptible gene of schizophrenia.

Objective To study the expression of retinoblastoma (Rb) protein in patients with breast cancer and its significance. Methods The expressions of Rb protein were detected in 52 patients with breast cancer, 20 patients with breast fibroadenoma or breast cystic hyperplasia (control group) by S-P immunohistochemical method. Results The negative rate of Rb expression in breast cancer (76.91%) was higher than that in control group (15.00%)(P<0.01).There was no relationship between the negative rate of Rb expression and the clinical pathological parameters in the patients with breast cancer (P>0.05). Of all 46 follow-up patients with breast cancer, the peritoneal recurrence and metastatic rate was 10.53% in 38 cases of the negative expression of Rb protein, and while the rate was 0 in 8 cases of the positive expression of Rb protein, there was significant difference (P<0.01). Conclusion The negative expression of Rb protein may be related to the initiation of breast cancer; as an tumor suppressor genes product,Rb protein may be considered as a new clinical indicator for monitor of breast cancer and provide information for adjuvant therapy.

Objective To evaluate the anti-ischemic effect of isosorbide dinitrate (ISDN) quantitatively. Methods 36 patients were divided into three group according to the different time points after the start of ISDN infused intravenously：groupⅠ (30 min), group Ⅱ(60 min), group Ⅲ(150 min).Tc-MIBI myocardial perfusion SPECT images was used to evaluate directly the anti-ischemic effect of ISDN on 36 patients with coronary heart disease. The changes in the percentage of ischemia area in entire left ventricular area and ECG were determined by the unfolded surface mapping before and after ISDN infusion. Results Tc-MIBI myocardial perfusion SPECT images were acquired, and reconstruction of the bull′s eye map and unfolded surface mapping were performed according to the dates of tomography images. In groupⅠ: at the 65% threshold value, (33.01±5.35)% at baseline,(28.9±5.23)% after ISDN was infused (P<0.05); at the 55% threshold value, (22.06±5.58)% at baseline,(19.60±4.07)% after ISDN was infused (P<0.05); the sum of ischemia ST segments was 60 at baseline, 51 after ISDN was infused. In group Ⅱ:at the 65% threshold value, (29.20±5.08)% at baseline, (20.81±4.16)% after ISDN was infused (P<0.001); at the 55% threshold value, (21.2±5.49)% at baseline,(17.52±5.59)% after ISDN was infused (P<0.001) ; the sum of ischemia ST segments was 58 at baseline, 47 after ISDN was infused. In group Ⅲ: at the 65% threshold value, (32.87±6.46)% at baseline, (20.81±4.16)% after ISDN was infused (P<0.001); at the 55% threshold value, (18.42±5.17)% at baseline,(14.18±3.61)% after ISDN was infused (P<0.001) ; the sum of ischemia ST segments was 64 at baseline, 41 after ISDN was infused. Conclusion The unfolded surface mapping of Tc-MIBI myocardial perfusion image can be used as a method of quantitative evaluation of the anti-ischemic effect of drugs and ISDN iv drip can improve the blood flow in myocardium and myocardial perfusion.

Objective To detect the expression of protein kinase C (PKC) in multidrug resistant cells of colorectal carcinoma after ionizing irradiation and observe the effect of PKC on occurrence and development of the multidrug resistant. Methods The effect of PKC on multidrug resistant HCG-8 cells of colorectal carcinoma after treated with X-ray was detected by indirect immunofluorescence technique and flow cytometry. Results Compared with sham-irradiation group, the positive per centage of PKC was increased significantly (P<0.05) after HCT-8 cells were treated with 2 Gy X-ray,the positive percentages of PKC were increased insignificantly after HCT-8 cells were treated with low dose (50,100 mGy) before high dose of irradiation, the increasing of positive percentage of PKC in the group of 2 00 mGy+2 Gy was significant(P<0.05).Compared with the group only treated with high dose of irradiation, the positive percentage of PKC in the group of 100 mGy+2 Gy was decreased significantly (P<0.05). Conclusion High dose of ionizing irradiation can increase the expression of PKC in colorectal carcinoma significantly, reversely when HCT-8 cells are pretreated with low dose before high dose there is no additional increase of PKC expression.

Methods The levels of D-Dimer and tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) were detected in 92 middle-aged patients with nonvalvular atrial fibrillation (AF group) and 60 patients with sinus rhythm (control group) by immune turbidimetry and enzyme linked immunoadsorbent assay (ELISA). Univariate analysis was used to determine the ifferences between two groups, and covariance analysis was used to determine the factors which might affect coagulation and fibrinolysis indexes. Results ①The plasma levels of D-Dimer ［(0.16±0.10）mg·L^-1］ and t-PA ［(42.58±30.28）μg·L^-1］ and PAI-1 ［(86. 03±21.43)μg·L^-1］ in AF group were significantly higher than those in the control group ［(0.10±0.08)mg·L^-1，(26.02±13.84)μg·L^-1， (64.94±24.35)μg·L^-1］(P<0.05 or　P<0.001). The ratio of PAI-1 /t-PA in AF group was higher than that in control group slightly. ②After adjustment of the factors which included sex, age and plasma reatinine, uric acid, blood sugar, triglyceride and cholesterol, the levels of D-Dimer（P=0.047）, t-PA（P=0.264）and PAI-1（P=0.001）in AF group were higher than those in the control group. Conclusion The middle-old aged patients with nonvalvular atria l fibrillation lose their balance of coagulation and fibrinolysis in the state of hypercoagulated and hypofibrinolysis.

Objective To investigate the genetic association between polymorphism of mdm2 gene and symptoms and pathological types of non-small cell lung cancer (NSCLC). Methods Polymerase chain reaction(PCR) and restriction fragment length polymorphism(RFLP) was used to identify mdm2 genotypes. The Pearson Chi square test and Woolf statistic method were used to analyze the relative risk and 95% confidence interval (CI) in order to find the genetic association between polymorphism of mdm2 gene and symptoms and pathological types of NSCLC. Results In the SNP rs1196337 (a G to A base change) AA genotype showed association with cough of NSCLC (P<0.05). Conclusion The polymorphism of mdm2 gene may be associated with symptom as cough of NSCLC.

Objective To investigate the expressions of prostatic specific antigen (PSA) mRNA in mononuclear cells of the peripheral blood in prostate cancer patients and controls and discuss its clinical significance. Methods 39 patients with prostate cancer and 12 healthy men were selected (normal control group). 5 mL anticoagulant blood was collected in every individual. The expressions of PSA mRNA in the mononuclear cells of the peripheral blood were measured by RT-PCR after delamination of the mon oclear cells. Results ①26 of 39 (66.7%) patients had positive PSA mRNA expression, in which (23 cases) serum PSA＞20.0 μg·L^-1, positive rate of PSA mRNA expression was 87.9% (20/23), while those serum PSA≤20.0 μg·L^-1, positive rate of PSA mRNA expression was 37.5% (6/16). ②In the 6 cases whose PSA mRNA was positive and serum PSA ≤20.0 μg·L^-1, the distribu tion of the biopsy Gleason were 1-4 score 0/3，5-7 score 1/8，8-10 score 5/5, pathologic findings revealed middle and low differentiation carcinoma in 6 patien ts with positive expression of PSA mRNA. Conclusion The detection of PSA mRNA has high sensitivity in prostate cancer patients whose serum PSA ＞20.0 μg·L^-1, which is a reliable method. The detection of PSA mRNA in mononuclear cells of peripheral blood can predict the micrometastasis in prostate cancer patients whose PSA≤20.0 μg·L^-1.

Objective To investigate the expressions of C-erbB-2, Nm23 and MDR1 in primary breast cancer and their relations with metastasis and prognosis. Methods Immunohistochemical method (S-P method) was used to detect C-erbB-2, Nm23, MDR1 expressions in 49 patients with primary breast cancer. The correlations between the three genes and their relations with metastasis and prognosis were analysed. Results The metastasis rate was higher in patients with high-expressions of C-erbB-2, MDR1 and low-expression of Nm23（P<0.05）. The expressions of C-erbB-2 and MDR1 had correlation (r_s=1.00, P<0.05). The expression of C-erbB-2 and Nm23 had no correlation（r_s=0.80，P>0.05）. The expression of MDR1 and Nm23 had no correlation（r_s=0.80，P>0.05）. The metastasis rate in patients with both C-erbB-2(+) and MDR1(+) was higher than the others. Their prognosis was negative. The metastasis rate in patients with both C-erbB-2(+) and Nm23(-) was higher than the others. Conclusion There are correlation between C-erbB-2, Nm23, MDR1 expressions and metastasis of breast cancer. Multi-predictors are better guide to patients than single predictor.

Objective To investigate the expression of of cyclooxygenase-2　(COX-2) in human gastric carcinoma and to identify the significance of COX-2 expression. Methods COX-2 protein expressions were deteced in 12 nomal gastric mucosa　and 40 gastric carcinoma tissues by immunohistochemistry.Results The expression of COX-2 in gastric carcinoma tissues (87.5%) was significantly higher than that in normal gastric mucosa (25.0%) (P<0.05). The expression of COX-2 in Ⅲ+Ⅳ stage of gastric carcinoma (96.55%) was higher than that in Ⅰ+Ⅱ stage (63.64%) (P<0.05); the COX-2 expression in lymph node metastasis group (96.43%) was higher than that in non-lymph node metastasis group (66.67%) (P<0.05). Conclusion The expression of COX-2 is related to TNM stages and lymph node metastasis. The expression of COX-2 maybe take part in the progress of gastric carcinoma and related to invasion,metastasis and prognosis.

Objective To study the protective effects of antipsychotic drugs Venlafaxine and Olanzapine on pheochromocytoma (PC12) cells after serum withdrawal. Methods The cells were cultivated in serum-free medium. Venlafaxine and Olanzapine were added in the medium.Morphological observation, MTT assays and LDH assays were used to study the protective effects of Venlafaxine and Olanzapine. Results Venlafaxine and Olanzapine prevented PC12 cells from the damage in serum-free medium, elevated PC12 cell survival rates,reduced the LDH release and kept cell membrane intact. The greatest effects were observed at concentrations of 100 μmol·L^-1 Olanzapine and 20 μmol·L^-1 Venlafaxine (P<0.01) and these effects were significantly different from their respective control groups and at 48 h the PC 12 cells had the greatest cell viabil ity.Conclusion Venlafaxine and Olanzapine can prevent PC12 cells from being injured after serum withdrawal. 100 μmol·L^-1 Olanzapine and 20 μmol·L^-1 Venlafaxine have the greatest protective effects and at48 h these drugs have greater protective effects than other groups.

Objective To explore the significance of serum VEGF-A and VEGF-C levels in relation to conventional clinopathological parameters of gastric carcinoma, investigate the relationship between serum VEGF-C level and expression of VEGF-C in carcinoma tissue. Methods 28 patients with gastric carcinoma and 21 healthy controls were included in the current study. Serum VEGF-C and VEGF-A levels were assessed by enzymed-linked immunosorbent assay（ELISA）. The expression of VEGF -C protein in gastric carcinoma tissue from 28 patients who underwent curative gastrectomy was examined by immunohistochemistry. Results Serum VEGF-C and VEGF-A levels were higher in patient s with gastric carcinoma than those in the healthy controls(P<0.01,respectively).The serum VEGF-C level showed no differenc in two groups: VEGF-C positive expression group and VEGF-C negative expression group (P<0.05). Serum VEGF-A level was correlated with lymph node metastsis (P<0.01),distant matastasis(P<0.01) ,TNM stage (P<0.01) and depth of invasion (P<0.01); Serum VEGF-C level was correlated with lymph node metastasis(P<0.05),TNM stage (P<0.05) and depth of invasion(P<0.05). Conclusion The serum levels of both VEGF-A and VEGF-C have potential usefulness as biologic markers of gastric carcinoma；Preoperative serum VEGF-C and VEGF-A levels are potentively useful as a marker reflecting lymph node status in gastric carcinoma；there is no relationship between serum VEGF-C level and expression of VEGF-C in gastric carcinoma.

Objective To explore the relationship between recurrent spontaneous abortion (RSA) and anti-β₂-glycoprotein Ⅰ(anti-β₂-GP Ⅰ) antibody. Methods The levels of anti-β₂-GP Ⅰ antibody in serum from 81 RSA patients and 39 normal women were detected by ELISA. Results The positive rate of anti-β₂-GP Ⅰ in RSA patients (42.0%) was obviously higher than that in normal women (7.7%) (P<0.01); there was obvious difference of the positive rate of anti-β₂-GP Ⅰ IgG in statistics between RSA patients (40.8%) and normal women(7 .7%)（P<0.05）, and there was no difference of the positive rate of anti-β₂-GP Ⅰ IgM in statistics between RSA patients and normal women (P>0.05). There was no difference of the positive rate of anti-β₂-GP Ⅰ in statistics between early and late, as well as between 2 times and more than 2 times abortions of RSA (P>0.05). Conclusion The anti-β₂-GP Ⅰ antibody is related to RSA, and it may be regarded as a immunological assistant diagnostic index for RSA.

Objective To isolate and identify the proteins responsible for aggressive character of giant cell tumor of bone (GCTB) with proteomic techniques. Methods 5 cases of aggressive GCTB and 4 cases of benign GCTB w ere selected as experimental and control groups, respectively, the differential protein expression were determined by isoelectric focusing / SDS acrylamide gel two-dimensional electrophoresis and Coomassie brilliant blue staining, then the differentially expressed proteins were identified by mass spectrographic analysis. Results(506±23) protein stains were found in experimental group, (468±28) protein stains were found in control group, there was significant difference between the two groups on two-dimensional electrophoresis (P<0.05 ), up-regulation of Annexin A1 and 14-3-3 protein ε, down- regulation of Peroxiredoxin 1 were found in experimental group. Conclusion The down-regulation of Peroxiredoxin 1 may be responsible for the aggression of GCTB, the up-regulation of Annexin A1 and 14-3-3 protein ε may be due to the result of reaction of organism, these three proteins may be selected as tumor markers of the aggressive GCTB.

Objective To explore the linearic regularity of airways′ spasm and spasmolysis in asthmatic patients, and provide theory bases for clinic treatment.　Methods After regular bronchodilation test, the pulmonary funct ion in 16 asthmatic patients and 14 volunteers were examined 15 min, 30 min, 1 h, 2 h and 4 h later, respectively. The indexes included forced vital capacity (FVC), forced expiratory flow in one second (FEV₁), peak expiratory flow (PEF), maximal midexpiratory flow (MMEF), expiration of 50% FVC (V₅₀) and expiration of 75% FVC (V₂₅) . Results 2 h after bronchodilation test, the big airways dilated completely P<0.05）,while the small airways dilated much later and incom plete except V₅₀. The percentage of indexes including FVC, FEV₁, PEF whose slopes of model straight line were negative was lower. But it was higher to indexes including MMEF, V₅₀ and V₂₅（P<0.05）. Conclusion The big airways dilate first and small ones later af ter given salbutamol. It is helpful for big airways spasm patients to choose the long-acting drugs or short-acting drugs, and for small airways spasm patients the long-acting drugs should be chosen.

Objective To study the histopathological changes and biological behavior of bowenoid papulosis and its relationship with apopotosis. Methods 20 bowenoid papuloses and 5 normal skin tissues were examined by DNA nick end labeling (TUNEL) technique and the pathological changes of bowenoid papulosis were observed by Hematoxylin-eosin-stained. Results Bowenoid papulosis exhibited disordered maturation and scattered,large,highly atypical cells mitoses throughout all layers of epithelium; and there were few apoptotic cells in 5 normal skin tissuese and many apoptotic cells in bowenoid papuloses. The apoptotic index （27.1±15.1） in bowenoid papulosis patients was significantly higher than that in control group (6.8±3.5) (P<0.01). Conclusion Benign biologic behavior of bowenoid papulosis may have relationship with increase of apoptotic cells.

Objective To study the histopathological changes and biological behavior of bowenoid papulosis and its relationship with apopotosis. Methods 20 bowenoid papuloses and 5 normal skin tissues were examined by DNA nick end labeling (TUNEL) technique and the pathological changes of bowenoid papulosis were observed by Hematoxylin-eosin-stained. Results Bowenoid papulosis exhibited disordered maturation and scattered,large,highly atypical cells mitoses throughout all layers of epithelium; and there were few apoptotic cells in 5 normal skin tissuese and many apoptotic cells in bowenoid papuloses. The apoptotic index （27.1±15.1） in bowenoid papulosis patients was significantly higher than that in control group (6.8±3.5) (P<0.01). Conclusion Benign biologic behavior of bowenoid papulosis may have relationship with increase of apoptotic cells.

Objective To study the diagnostic value of MR mammography （MRM）　in judging the nature of breast disease. Methods 145 cases with breast disease were retrospectively studied to investigate the relationships between two parameters (early-phase enhancement rate and the type of the delayed enhancement) and different breast diseases. All patients underwent MRM examination before surgical operation. Results ①Strengthen multiple in early stage larger than 3 times was inclined to malignant lesion; Strengthen multiple smaller than 1 time was inclined to benign lesion. ②Drop type of delayed enhancement was the important parameter in diagnosis of ductal carcinoma. If drop type of delayed enhancement was viewed as the diagnosis standard of the ductal carcinoma of breast onl y:81% cases with invasive type of the ductal carcinoma, 65% cases with noninvasive type of the ductal carcinoma had been diagnosed correctly; 14% cases with benign breast lesions had been diagnosed exaggeratedly. ③If strengthen multiple in early stage larger than 2.5 times in rising type of delayed enhancement and drop type of delayed enhancement was viewed as the diagnosis standard of the ductal carcinoma of breast,87.9% cases with invasive ductal carcinoma, 73.1% cases with noninvasive ductal carcinoma had been diagnosed correctly; 21.4% cases within benign breast lesions had been diagnosed exaggeratedly. Conclusion Compared with differential of disease nature by the type of the delayed enhancement only, the correct rate of diagnosis of breast ductal carcinoma mentioned above is raised if two parameters are combined. But the correct diagnosis rate of benign lesions is decreased.