Objective To study the inhibitory effects of gene combined radiotherapy on mice transplanted with B16 melanoma.Methods Alkaline lysis assay was used to extract and purify the plasmid. Plasmid DNA was injected into tumor by microinjection assay. Mice were inoculated with 5×10⁵ of B16 melanoma in right hind legs and the therapy was performed when the diameter of tumor reached at 0.3-0.5 cm. pNE-mIL-12 and pcDNA-B7-1 plasmids were injected locally three times following with irradiation three times. The tumor growth rate of mice was observed. Results The anti-tumor effect of pNE-mIL-12 combined with pcDNA-B7-1 plasmid following with 2 Gy X-ray irradiation was much better than other groups. It showed that the tumor growth rate was slowed, the survival days of mice were delayed significantly (P<0.01,P<0.001), the death rate of mice 31 d after therapy was only 22.2% and the tumor was deleted in one mouse. Conclusion Multi-genes combined with radiotherapy could inhibit the tumor growth of mice inoculated B16 melanoma and the inhibitory effect is better than single gene therapy and radiotherapy.

Objective To study the effects of whole-body irradiation（WBI）with different doses of X-rays on apoptosis in mouse splenocytes and peritoneal macrophages. Methods The apoptosis percentages of mouse splenocytes and peritoneal macrophaes were detected with flow cytometry（FCM）at different time after the whole-body X-irradiation using the staining of Annexin-V and PI. Results As com pared with the control，the percentage of apoptosis in mouse splenocytes began to increase gradually 24 h after WBI with 2 Gy X-rays（Ｐ<0.05），and the percentage of apoptosis in mouse peritoneal macrophages aros e 4 h after 2 Gy irradiation,and kept higher level till 48 h（Ｐ<0.05，Ｐ<0.01，Ｐ<0.001）. After WBI with 0.075 Gy X-rays，the percentage of apoptosis in mouse splenocytes reduced at 24 h （Ｐ<0.05），and the percentage of apoptosis in peritoneal macrophages significantly reduced from 2 h to 16 h（Ｐ<0.05）as compared with the control. Conclusion High dose irradiation can stimulate apoptos is in mouse splenocytes and peritoneal macrophages， while low dose irradiation can cause an opposite effect and even suppress immunocyte apoptosis.

Objective To explore the expressions of proliferating cell nuclear antigen (PCNA), IL-1β and IL-6 in tumor derived from injection of tumor strain cells, determine the relationships between the expressions of the above cytokines an d clinical pathological parameters. Methods Immunohistochemical methods were employed to detect the expressions of PCNA,IL-6 and IL-1β in tumor derived from mou se hepatocarcinoma and colon cancer cells. Results PCNA highly expressed in nuclear and IL-6 and IL-1β expressed in tumor mesenchymal vessals. The expres sions of PCNA and IL-1β were significantly related to the tumor lymph node metas tasis and tumor weight. Conclusion The expressions of PCNA,IL-6 and IL-1β in tumor tissues are identical and closely related to pathological parameters of lymph node metastasis and tumor weight.

To explore the effects of fluoride with different doses on the expressions of TGF-β and Smad2/3 in fibroblast and osteoblast at different periods. Methods Fibroblasts and osteoblasts were exposed to different concentrations of fluoride (0，0.0001，0.001，0.1，1，10 and 20 mg•L^-1) . The levels of TGF-β protein and Smad2/3 at 2,4,24,48 and 72 h after treatment were measured by using ELISA method. The expression of TGF-β mRNA was tested with RT-PCR method. Results In fibroblasts，the contents of TGF-β protein were decreased in the groups of 0.001，0.1，1，10 and 20 mg•L^-1 F^- a t the time of 2 h and in the groups of 0.0001，0.001，0.1，10 and 20mg•L¹ at the time of 4 h（P<0.01，P<0.05). Thelevels of TGF-β mRNA were increased in the groups of 1 and 20 mg•L^-1F^- at 48 h and the expressions of Smad2/3 were increased in fluoride groups during 2 to 24 h. Inosteoblasts，the expressions of TGF-βprotein and mRNA were increased significantly in the groups of 0.0001，0.001 and 1 mg•L-1 F^-at 48 h（P<0.01，P<0.05)and the level of Smad2/3 elevated in the group of 20 mg•L^-1 F^- at 48 h（P<0.01)；Conclusion TGF-β may play roles in the develop ing of osteogenesitic function in fibroblasts induced by fluoride.

Objective To observe the action of lowering blood fat of Agaricus blazei Murill ω-6 polyunsaturated fatty acid(ω-6APFA) on hyperlipemia rats and mice. Methods Sixty rats were randomly divided into control group, model group, natural soybean phospholipids capsule (NSPC)0.70 g•kg^-1 group,ω-6APFA 0.28, 0.14, 0.07 g •kg^-1 groups(n=10). The other groups except control group were given high-fat diets for 14 d, on the fourteenth day the rats were administered orally, the control group and model group were administered distilled water 10mL•kg^-1 at the same volume , 14 days after continuous administration ,rats were anesthetized , the blood were extracted from abdominal artery ，T-CHO, TG, HDL-C, LDL-C in sera were determine d. At the same time, the activity of SOD in liver and the content of MDA were determined, the fat accumulated coefficient was calculated. 72 male mice were randomly divided into control group , model group, NSPC 1.0 g•kg^-1 group,ω-6APFA 0.4, 0.2, 0.1 g•kg^-1 groups(n=12) . Mice were administered continuously,16 h before the last administration ,except control group, the mice in the other groups were injected 75% yolk physiological salt solution 0.5 mL through the abdominal cavity, and began to starve ,1 h after the last administration, blood was extracted from eyeball , serum T-CHO and TG were determined. Results Compared with model group, T-CHO and TG in rats treated with NSPC 0.70 g•kg^-1 and ω-6APFA 0.28 g•kg^-1 all reduced and HDL-C raised obviously(P<0.001), the activities of liver SOD in rats treated with NSPC 0.70 g•kg^-1 and ω-6APFA 0.28,0.14 g• kg^-1 obviously raised (P<0.05, P<0.001, P<0.05), MDA reduced obviously(P<0.01,P<0.001，P<0.01), fat accumulation coefficient in rats treated with NSPC 0.70 g•kg^-1 and ω-6APFA reduced obviously (P>0.01). Compared with model mice , T-CHO and TG in acute hyperlipemia mice treated with NSPC 1.0 g•kg^-1 and ω-6APFA0.4,0.2 g •kg^-1 reduced obviously(P<0.05, P<0.01,P<0.05).Conclusion ω-6APFA has the function of lowering blood fat on hyperlipemia rats and mice.

Objective To study the anti-tumor activity of Phellinus linteus(PL) and itsimmunomodulational effects. Methods Solid tumor was induced by injecting HepA, MFC, S₁₈₀ cells (310⁶cells/animal) subcutaneously to the right anterior limb in mice. The tumor weight, thymus weight and spleen weight were measured after mice were executed. The effects of PL on lymphocyte proliferation, mitogen-induced proliferatoin of ICR mouse splenocytes were measured by MTT. Meanwhile, lymphocytes were prepared as described above. SRBC (10%, 0.2 mL) was injected to abdomina l cavity to elevate the level of antibody production by B cells. Spleen cells suspension (1×10⁷•mL^-1) was used as the effect cells and L₉₂₉ cells was used as the target cells. L₉₂₉ cells engulfing neutral red determined the cytotoxic act ivity of NK. Simultaneously, phagocytosis of mice was detected using the method that macrophages engulfed neutral red. The effect of PL on the production of antibody was studied, the lymphocytes from mice wereprepared as described above. Results The high-dose (2.0 g•kg^-1) and mid-dose (1.0 g•kg^-1) of PL could significantly inhibit the growth of HepA, MFC and S₁₈₀tumor, the inhibitory rates were 38.19% and 37.15%,32. 46% and 30.91%,36.0% and 34.16%, respectively (P<0.05, P<0.01). The average absorbanc e of lymphocytes in 2.0 g•kg^-1 group was higher than that in the untreated control group. The numbers of antibody forming cells were increased by PL treatment at doses of 2.0 g•kg^-1 and 1.0 g•kg^-1 (P<0.05, P<0.001), the difference was significant compare d with control group (P<0.05). PL (2.0 g•kg^-1) enhanced the cytoto xic activity of NK cells (P<0.01). Average weights of the spleens and thymus (0.1 mg per gram body weight) were not significantly greater than those in positive control groups . PL(2.0 and 1.0 g•kg^-1) increased the activity of macrophage (P<0.05, P<0.01). Conclusion PL has anti-tumor activity and immunomodulational effect and probably inhibit tumor by means of activating the immunocompetence of the body.

Objective To observe the changes of learning and memory abilities induced by cumulative lactic acid for long time in atlas dentata vertebrae in mice. Methods 30% lactic acid was injected into the place between the first and the second cervical vertebrae. The changes of lesarning and memory in mice were observed with step-down avoidance test,water-maze test and hole-board test. The organ index of the brain was calculated and the pathological changes were observed. Results In step-down avoidance test, the reaction period prolonged and wrong times increased obviously(P<0.01), and the latency period shortened significantly. The whole-distance swimming time prolonged and mistakes increased in water-maze test(P<0.01). In hole-board test, the latency period prolonged and the times of extending the holes decreased significantly(P<0.01). The organ index of model mice decreased significantly(P<0.01), and pathological changes such as the collo id corpuscle, necrotic cells increasing, inflammation cells infiltrating in blood vessels were discovered. Conclusion The abilities of learning and memory of the model mice are hurtsignificantly by accumulation of lacticacid in cervical part.

Objective To study the relationships between antiespilepsy action of zinc henytoin and contents of glutate,aspartate,glycine andγ-GABA in mouse brain. Methods One hundred and fifty Kunming mice were randomly divided into two groups, seventy-five mice in every group, one group was normal , the rats in another group were built epilepy models by the maximal electroshock seizure test (MES). The normal and epileptic mice were randomly divided into five groups,respectively:gummy power, sodium phenytoin 15 mg•kg^-1and 50 mg•kg¹, zinc phenytoin 15 mg•kg ^-1 and 50 mg•kg^-1 , fifteen mice in every group , the mouse brain was took out after the normal mice were given drugs for 3 weeks. After given drugs for 30 min , the epileptic mice were induced epileptic outbreak by MES again, the mouse brain was took out after 30 min . The contents of four kinds of amino acids in normal and epileptic mouse brain were determined by pre-column derivation of high performance liquid chromato graphy with 2,4-dinitroch lorobenzene. Results Compared with gummy power groups, sodium phenygoin and zinc phenytoin in low dosage(15 mg•kg^-1) ha d no significant difference of the contents of four kinds of amino acids between normal and epileptic mouse brain.Zinc phenyt oin and sodium phenytoin in high dosage(50 mg•kg^-1) all obviously increased the contents of γ-GABA and glycine in normal and epileptic mouse brain, while they hadslight effects on the other two kinds of amino acids. Conclusion The antiepilepsy fu nction of zinc phenytoin is related to increasing inhibitory neurotransmitter γ-GABA and glycine.

Objective To observe the effect of chronic variable stress on the forced swimming test of rats, and investigate its preliminary mechanism. Methods First a mark was given to each Wistar rat by forced swimming test. 12 Wistar rats which had similar scores were selected and divided into control group and model group averagely. The behavior changes of rats were induced in the forced swimming test by chronic stress and the plasma cortisol level was analyzed through radioimmunoassay (RIA) methods. The changes of hippocampus CA3structure were observed with microscope. Results Compared with control group, chronic variable stress increased immobility time, reduced climbing time evidently and enhanced the weight of adrenal gland and the level of plasma cortisol (P<0.01). The number of hippocampus CA3 cells was decreased (P<0.05). Conclusion Chronic variable stress can induce behavioral despair of rats and this effect relates to the excessive secretion of cortisol and the damage of hippocampus CA3.

Objective To amplify the enamel matrix serine proteins 1(EMSP1) in vitro and to establish gene targeting vector and transgenic mouse models to study the physiological and pathological function of EMSP1.Methods Two pairs of primers were d esigned by Oligo 6.0 software and synthesized according to EMSP1 gene sequence. Two gene fragments of 129 strain mouse EMSP1 were amplified from mouse genomic DNA by PCR-5 000 bp 5′ arm and 1 700 bp 3′arm. The two arms were cloned into a universal gene knockout vector-pSSC-9 respectively on either side of the positive selective marker neo. The targeting vector was identified by PCR， restriction endonuclease and sequence analysis. Results The restriction endonuclease identifiation result indicated that 5 000 bp and 1 700 bp fragments were obtained and homogeneous long and short arms were cloned into vector.DNA squence analysis showed that EMSP1 gene targeting vector pSSC-9-EMSP1 was successfully constructed. Conclusion The mouse EMSP1 gene targeting vector is constructed successfully.

Objective To explore the influences of different inductive methods on cartilage repair by tissue engineered cartilage with the seed cell of mesenchymal stem cells. Methods Rabbit mesenchymal stem cells were divided into dexamethasone-inducing group and TGF-β₁-dexamethasone co-inducing group. The cartilage defects were repaired by autologous tissue engineered cartilage constructs. The defects of control group were filled with scaffold without cells. Specimens were harvested 6 and 12 weeks postoperatively and assessed by histological grading and in situ detection of apoptosis. Results The repair tissue formed perpendicurar column structure resembling that of normal cartilage in TGF-β₁-dexamethasone co-inducing group. The histological score 12 weeks postoperatively was higher in co-inducing group (20.26±1.35) than those in dexamethasone-inducing group (14.52±1.46) and control group (4.12±1.13). But the formation of tidemark was not observed in the repair tissue. Cells of repair cartilage at the bone-car tilage interface showed apoptosis. The ratios of apoptosis in dexamethasone-inducing group were (21.4±4.5) six weeks postoperatively and (7.3±2.2) twelve weeks postoperatively. The ratios of apoptosis inTGF-β₁-dexamethasone co-inducing group were (19.8±4.7) six weeks postoperatively and (6.9±2.0) twelve weeks postoperatively. The ratios of apoptosis at 6 th week were higher than those at 12 th week postopera tively with statistical significance(P<0.05). Conclusion The TGF-β₁ and dexamet hasone co-inducing group shows better repair of the cartilage defect and more slight and later degeneration of the repair tissue. The degeneration of cartilagetissue engineering transplant is correlated with the inducing environment prior to transplantation, apoptosis, and the subchondral vascular system.

Objective To evaluate the osteoinductive activity of recombinant adenoviral vector carrying the human BMP-2 gene (AdBMP-2) and investigate the mechanism of bone formation induced by the bone morphogenetic protein 2 (BMP-2). Methods AdBMP-2 infected rabbit bone messenchymal stem cells (BMSC) were cultivated in vitro. The expression of hBMP-2 was determined by in situ hybridization and im munohistochemical analysis, and ALP wasdetermined by enzyme cytochemistry 7 d after infection. At 21th day, Alizarin red S staining was used to evaluate calcium-rich deposits by cells transfected in culture. 30 μL AdBMP-2 solution (2×10¹pfu•mL^-1) was injected into the muscle of the left hind limb of the nude rat, and 30 μL Adβ gal solution (2×10^1＊pfu•mL^-1) w as injected into the muscle of the right hind limb as control. Animals were sacrificed at 20th day following injection to investigate the bone formation histologically and the expression of BMP-2 in the specimens of the injection sites by immunohistoch emistry and in situ hybridization method. Results The expressions of BMP-2 and ALP in the transduced BMSC were confirmed at 7 th day after infection. A deeply stain ed mineralized layer was evident at 21th day in transduced cells. In vivo experiment, BMP-2 gene therapy induced mesenchymal monocytes chemotaxis, proliferation,differentiation into chondrocytes, with subsequent endochondral bone formation. At the same time, hBMP-2 expression was observed in the chondrocytes, osteoblasts and mesenchymal monocytes. Conclusion AdBMP-2 posseses the osteoinductive activity in vitro and in vivo.

Objective To clone the heat shock protein 70 (HSP70) gene of mycobacterium tuberculosis bovis,and construct prokaryotic plasmid of mycobacterium tuberculosis HSP70 (Mtb HSP70) and express it in E. coli,further more,to obtain high pure Mtb HSP70. Methods The whole Mtb HSP70 DNA sequence was amplified from mycobacterium tuberculosis bovis genome by polymerase chain reaction (PCR).Then the sequence was cloned into plasmid pGEM-T easy and sequenced. The recombinant expression plasmid PQE30-Mtb HSP70 was constructed and expressed in E.coli M15,and Mtb HSP70 was purified by Ni-NTA affinity chromatography column. Results The sequence of Mtb HSP70 was amplified and identical with that published in GenBank. The prokaryotic expresstion plasmid of Mtb HSP70 was constructed success fully.With induction of IPTG, a new fusion protein with relative molecular mass of 70 400 was expressed and mainly located in inclusion bodies,the expressed 6×his-Mtb HSP70 fusion proteins were identified by Western blotting with anti-His monoclonal antibody. Conclusion A  confirmed Mtb Hsp70 gene is cloned and expressed in E. coli successfully,and high pure Mtb HSP70 is obtained.

Objective To evaluate the effect of hepatocyte growth factor(HGF) on transvascular metastasis of sarcoma cells. Methods The models of intravascular and extravascular migration of tumor cells in vitro were used to observe transvascul ar metastasis of sarcoma cells(HT1080). Results HT1080 migrated through basement membrane into blood vessels,and migrated through a gap between adjacent endothelial cells into extracellular matrix.The greater number of transmigrated HT1080 treated with HGF was observed(P<0.01).Conclusion HGF can stimulate transvascular metastasis of sarcoma cells as a motogen.

Objective To investigate the anti-tumor effects of combined recombinant murine interleukin-12 and GM-CSF gene adenovirus vector (AdCMVIL-12, AdCMVGM-CSF) on B16 melanoma. Methods B16 melanoma cells were injected subcutaneous at right back of C57/BL6 mice, 48 tumor bearing mice were divided into 4 groups at random, when tumor diameter attained to 5 mm, AdCMVIL-12,AdCMVGM-CSF and AdCMVIL-12+AdCMVGM -CSF and AdCMVLacZ (control) were injected in different groups, tumor growth and immune responses of each group were tested, and the IL-12 and GM-CSF levels in serum were observed. Results In combined group, necrosis, lymphocytes and macrophages were found in tumor tissues. 30 d after treatment, the average volume in combined group was （60.73±11.36）mm³, compared with AdCMVLacZ (control) group, and AdCMVIL-12 group, AdCMVGM-CSF group ［(675.18±80.59）,（186.91±19.15）,（223.45±23.11）mm³］, there were significant differences (P<0.01). Combined IL-12 and GM-CSF gene therapy could enhanced　anti-tumor immune response of B16 melanoma bearing mice and increased the expressing time of IL-12 and GM-CSF, and had very strong specific kill effect. Kill probability was （69.53±9.20）%. Conclusion Combined IL-12 and GM-CSF gene therapy can enhance the anti-tumor immune response of B16 melanoma bearing mice and decrease side -effect of using AdCMVIL-12.

Objective To study the relationship between CXXC motif and biologic activity of human augmenter of liver regeneration protein(hALRp). Methods The function of CXXC motif and its relationship with bioactivity were investigated between hALRp and hALRp-C65A groups by detecting the sulfhydryl oxidase activity； the interaction of target protein with GST-Na⁺,K⁺-A TPase fusion protein was examined by GST-Pulldown and MTT was used to study the ability of hALRp-C65A to promote HL-7702 hepatic cells proliferation in vitro. Results The turnover number (TN) of hALRp in sulfhydryl oxidase activity assay was 1.25±0.21, while TN was 0 of hALRp-C65A, there was significant difference between them(P<0.05). The interaction between GST-Na⁺,K⁺-ATPase fusion protein and hALRp-C65A still remained. And the ability to promote HL-7702 proliferation of hALRp-C65A didn't have marked change when compared with hALRp. Conclusion The CXXC motif is essential to the activity of sulfhydryl oxidase. But this motif contributes less to the interaction between hALRp and Na⁺,K⁺-ATPase and the ability to stimulate HL-7702 hepatic cell growth in vitro.

Objective To study the effects of Panaxadiol Saponins (PDS) on glycemia, lipid and the improvement of insulin resistance (IR) in type 2 diabetes mellitus rats (T2DM). Methods Rats were randomly divided into normal group, model group, PDS 35,70 and 140 mg•kg^-1 groups and metformin group. Rats were fed by high fat diet for 10 d and injected with small dose of streptozotocin(STZ) 55 mg•kg^-1. Rats with plasma glucose over 11.1 mmol•L^-1we re assigned to T2DM model group,continually fed with PDS 35,70,140 mg•kg^-1•d-1 for 4 weeks and the values of fasting blood glucose (FBG) were assayed every week . Then FBG, Ins, C-peptide, FFA, TC, TG, HDL-c, LDL-c, MDA content and SOD activity were measured after 4 weeks. Results After treated with PDS for 4 weeks, the FBG in PDS 70, 140 mg•kg^-1 and metformin groups decreased significantly compared with normal group from the third week to the forth week (P<0.01); the liver glycogen　content was higher than that in model control (P<0.05 or P<0.01), the leve ls of TC, TG, FFA, TC/HDL-c, LDL-c/HDL-c and MDA were lower than those in model control (P<0.05 or P<0.01) . SOD activity, C-peptide content and ISI increased significantly (P<0.05 or P<0.01), while LDL-c, HDL-c changed little (P>0.05). Conclusion PDS could decrease the blood glucose, improve the disorder of the serum lipid and insulin sensitivity in T2DM rats.

Objective To study the effect of Acanthopanax senticosus saponins (ASS) on organ blood flow and vascular resistance in anesthetized dogs. Methods The cerebral blood flow(CBF), cerebral vascular resistance(CVR),peripheral blood flow(PBF),peripheral vascular resistance(PVR), blood pressure(SDP and DBP) and heart rate(HR) were measured using the MF-27 electromagnetic flowmeter. Results ASS (15 and 30 mg•kg^-1 iv perfusion for 60min) augmented CBF and PBF, decre ased CVR and PVR within 180 min and slowed down HR at 60-180 min. In addition, ASS (30 mg•kg^-1 iv perfusion for 60 min) could obviously reduce SBP and DBP . Conclusion Protective effect of ASS on experimental cerebral ischemia may be rela ted to improving cerebral circulation and increasing cerebral blood supplyment.

Objective To study the therapeutic effects of oleum curcumae wencho wensis on skin of guinea pigs damage irradiated by UVB and its mechanism. Methods The skin of guinea pigs was irradiated by UVB (28.38 J•cm^-2, 30 d) to establish the oxidative damage model. The skin erythema and the rough level were observed during the experiment; the appearance and thickness of epiderm and the number of fibroblast were observed under light microscope.The activities of GSH-Px, SOD, CAT and T-AOC and the content of MDA in the supernate of skin homogenate were detected with biochemical methods. Results The epiderm in UVB irradiated group and blank group thickened, the number of fibroblasts decreased;and the epiderm in therapeutic group didn't thicken, but the number of fibroblasts increased, but those in the unirradiated side of every group did not change. Compared with normal group, the contents of MDA in the supernate of skin homogenate in UVB irradiated group and blank group increased, but that in therapeutic group decreased. The activities of GSH-Px, SOD, CAT and T-AOC in UVB irradiated group and blank group decreased, but those in therapeutic group increased, and those in the unirradiated side of every group didn't change. Conclusion Oleum curcumae wenchowensis has the therapeutic effects on skin damage irradiated by UVB, which may be mediated by increasing the contents of antioxidases and eliminating the free radical.

Objective To observe the suppressive effects of ginsenoside Rg3 combined with X-ray irradiation on the growth of melanoma B16 cells. Methods C57BL/6J mice implanted with melanoma B16 cells were used as models. The volume of tumor in the mice in vivo and the survival rate of B16 cells in vitro treated with Rg3 combined with irradiation were observed. Results As compared with control, the tumor volume in the mice in the Rg3 group reduced significantly 6 d after im plantion with B16 cells (P<0.001); the volume in the irradiation group and the Rg3 plus irradiation group all reduced 12 d after irradiation, the latter reduced more significantly (P<0.05). Meanwhile，Rg3 combined with irradiation decreased significantly the survival rate of B16 cells as compared with the other 3 groups. Conclusion Ginsenoside Rg3 could suppress the growth of melanoma, B16 cells and increase its radiosensitivity

Objective To investigate the inhibitory effect of ginsenoside Rg3 on herpes simplex virus type 1(HSV-1) in vitro and its immunoregulative effect. Methods Crystal violet staining was used to observe the inhibitory effect of Rg3 on the cytopathic effect caused by HSV-1. MTT colorimetry was used to detect the influence of Rg3 on the proliferation of Vero cells and mouse splenocytes. The mouse splenocytes were induced with Rg3, and the activity of IL-2 was detected by lymphoblast proliferation assay and IFN-γ by cytopathic effect inhibition. Results The A₅₇₀ values of cytopathic effect in the experimental group were significan tly higher than those in the control group when doses of Rg3 were 1.56-25 mg•L^-1 in vitro(Ｐ<0.05).The A₅₇₀ values of proliferation effect on normal Vero cells and mouse splenocytes had no significant difference between the experimental group and control group when doses of Rg3 were lower than 25 mg•L^-1 (P>0.05).The secretary levels of IL-2 and IFN-γ in experimental group were significantly higher than those in the control group in vitro when doses of Rg3 were 3.13-6.25 mg•L^-1 (P<0.05). Conclusion Rg3 in some extent could inhibit HSV-1 in vitro and has no poisonous effect on normal Vero cells and mouse sple nocytes. Rg3 could significantly stimulate the secretion of IL-2 and IFN-γ, thereby participate immunoregulative effect.

Objective To examine the effects of YiXinTong pellet on the serum enzymes and ultrastructure of myocardium in canine model with acute myocardial infarction. Methods Thirty dogs were randomly divided into control group , YiXinTong tablet group, YiXinTong pellet 25,50 and 100 mg•kg^-1 groups. The left anterior de scending branches were ligated in anesthetic dogs to establish acute myocardial infarction models. The activities of AST, CK and LDH in serum were detected and the changes of ultrastructure of myocardium were observed. Results YiXinTong pellet decreased the activities of AST, CK and LDH（P<0.05，P<0.001）,compared with control group,there was no significant difference between YiXinTong tablet group and YiXinTong pellet groups(P>0.05). Ultrastructure of myocardium was nearly normal in YiXinTong pellet groups.Conclusion YiXinTong pellet shows apparently protective effects on cardiac muscle of experimental canine model with acute myocardial infarction. 

Objective To investigate the effect of Ast raglaus Memberanaceus (AM) injection on myocardial enzymes and ultrastructure of myocardium in dogs with acute cardiogenic shock . Methods Thirty dogs were divided randomly into control group , ShengMai(SM) group, AM 2.5，5.0，10.0 g•kg^-1 groups,six dogs in each group.An acute cardiogenic shock model was made by ligating left anterior descending branch in anesthetic dog. The levels of myocardial enzymes(AST,CK,LDH) in serum were detected and the changes of myocardial ultrastructure were observed .Results Compared with control group, the myocardial enzyme activities in serum were decreased significantly in AM groups（P<0.05，P<0.001）.There were no sig nificant differences between SM group and AM groups(P>0.05). Filaments arranged regularly and mitochodrias was normal of myocardium in AM 10.0 g•kg^-1 group. Conclusion AM can reduce ultrastructural injury of myocardium and protect cardiac muscle from hypoxic-ischemic injury.

Objective To observe the inhibitory effect of LaCl₃ on the growth of human hepatocellular carcinoma cell lines SMMC-7721 and its mechanism. Methods The cells were treated with different doses of LaCl₃ and the growth curve, colony inhibitory rates,the levels of AFP of cultured cells were detected at 1,3 , 5 and 7 days after they were dealed withLaCl₃. The changes of cell cycle were detected by flow cytometry and the expressions of cell cycle protein CyclinD₁,PCNA,CDK₄ and P16 were observed by immunocytochemistry staining . Results MTT test showed tha t SMMC-7721 cell proliterating activity was obviously inhibited by LaCl₃ in a time and dose related manner at concentrations of 0.50 and 1.00 mmol•L^-1. The colony forming inhibitory rate of 0.10, 0.50 and 1.00 mmol•L^-1 LaCl₃ were 51%, 67% and 97% （P<0.05, P<0.01 ）.Cell cycle progress of SMMC-7721 changed obviously when treated with different does of LaCl₃ ,the percent of G₀/G₁ phase was higher and S phase lower, the difference was significant compared with control group（P<0.01). 0.10,0.50 and 1.00 mmol•L^-1 LaCl₃ could decreased the levels of AFP, the effect of 1.00 mmol•L^-1 LaCl₃ was notable（P<0.01 ）.Immunocytochemist ry showed that LaCl₃ could enhance the expression of P16 protein and decrease the expressions of CyclinD₁,PCNA and CDK₄. Conclusion LaCl₃ could inhabit the growth of SMMC-7721 cell and its mechanism might be connected with the inhibiti on of cell cycle progress and decrease the degree of malignancy.

Objective To study the inhibitory effect of 3,3-Diindolylmethane (DIM) on PC3M cells. Methods Growth inhibition test was used to examine the changes of cell proliferation ,and flow cytometry was used to examine the changes of cell cycle and the apoptosis of human prostate cancer cell line PC3M cultivated in vitro. Results DIM of different concentrations could significantly inhibit the proliferation of PC3M cells. When the concentrations were 10,30,60,120 μmol•L ^-1 ,the inhibitory rates were 35.6%,58.8%,76.6%,90.5%,respectively. G ₁ phase was blocked, and apoptosis was induced in 22.6% PC3M cells. Conclusion DIM can inhibit the prolifera tion and cell cycle of PC3M cells and induce apoptosis.

Objective To study the effect of morphine on gene expression of enzymes involved in salvage and catabolism pathways of purine nucleotides metabolism in nerve cells. Methods PC12 cells were cultivated and divided into morphine treated groups which were treated with morphine (10 mg•L^-1 culture) for 12, 24, 48,72 h, and control groups which were treated with normal saline for 12, 24, 48 and 72 h. Total RNA of PC12 cells was isolated. HGPRT, AK, ADK mRNA levels were determined by using the reverse transcription polymerase chain reaction (RT-PCR);the β-actin mRNA expression was used as an internal control. Results As compared with corresponding control groups, HGPRT mRNA levels in PC12 cells were increased significantly after 12 and 24 h treatment with morphine(P<0.05), became lower after 48 h and became higher again after 72 h (P<0.05). AK mRNA levels in PC12 cells were decreased significantly after 12 and 24 h treatment with morphine(p<0.05), become higher after 48 h (P<0.05), and were normal after 72 h. ADA mRNA levels were decreased in morphine treated PC12 cells in all the given periods with significant difference in 12 h (P<0.05). Conclusion Morphine may affect purine nucleotide metabolism of nerve cells, increase the adenylate and adenosine levels, which may be one of the mechanism of morphine dependence and tolerance.

Objective To investigate the effects of topiramate on neurons apoptosis and the hydrous capacity in brain tissue in rats with perinatal acute ischemic brain damage. Methods The perinatal acute ischemic brain damage model was established by ligation of both uterine arteries of full-term pregnant Wistar rats(n=315). They were divided randomly into topiramate one and five times groups and hypoxic-ischemia (HI) group （n=105）, and normal collate group consisted of 105 normal Wistar rats .The effects of topiramate were observed by the changes of hippocampal apoptosis cells labeled in situ end TUNEL methods at different time spots (postnatal 3 h, 6 h, 24h, 3 d, 7 d, 14 d, 21 d, 28 d) and t he hydrous capacity in brain tissue within 7 d after birth.Results The hydrous capacity in brain tissue in administering drug one time group was remarkably less than that in HI group at 12 and 72 h after birth, and it in administering drug five time group was remarkably less than those in HI group and administering drug one time group at 48 and 72 h after birth（P<0.05）.The number of apoptosis neurons in administering drug one time group was remarkably less than those in HI group 24 h,3 d,7 d,21 d after birth, it in administering drug five time group was remarkably less than those in HI group and administering drug one time group 3 and 7 d after birth（P<0.05）. Conclusion Topiramate can remarkably lessen the degree of neurons apoptosis and shorten the duration of apoptosis; delay the occurrence of cerebral edema after HI, lessen the degree of cerebral edema, and shorten the duration of cerebral edema. The protective nerve function of topiramate in the many times administer ing drug group is obviously higher than that in the one time administering drug group.

Objective To detect the transfusion transmitted virus-like mini virus (TLMV) sequence gene in patients with chronic hepatitis and to study the TLMV infection in population. Methods Two sets of primers in the most conservative regions of Japan strains TLMV-CBD279 and CBD231 were designed to amplify TLMV templete extracted from sera of patients with chronic hepatitis B using PCR. PCR products were cloned into pGEMR-T vector and sequenced. Results The results showed that 72%of TLMV sequence isolated in China was identical to that in Japan, suggesting that there was TLMV infection in patients with chronic hepatit is B in China.The patients with hepatitis virus C and the healthy blood donor had the highest infectious rates of TLMV. Conclusion TL0MV infection exists in patients with chronic hepatitis B in China.

Objective To investigate the genotypes of hepatitis B virus in chronic liver disease patients in Changchun, China . Methods HBV DNA extracted from the 39 serum samples of chronic liver disease patients were amplified by nested polymerase chain reaction (nested-PCR) to obtain the S gene sequences, and all the PCR products were sequenced by dideoxy-chain-termination method. Results The S gene nucleotide sequence was acquired and the corresponding amino acid sequence was deduced.Compared with the consensus sequence,16 strains belonged to genotype C/adr and 7 strains to genotype C/adw; 16 strains belonged to genotype B/adw; and the amino acids of antigen determinant “a” of two strains changed in the 2 serum samples. There were many mutational positions and some of these caused amino acid residue substitution. No special point mutation was found which was similarly related to hepatocellular carcinoma. Conclusion In chronic liver diseases patients infected with HBV , S gene sequence of HBV has its variability.

Objective To analyze the HEV genetic structure and nucleotide homology isolated from sporadic acute hepatitis E in Changchun.Methods HEV RNA was detected with reverse transcription nested polymerase (RT-nPCR) and sequenced by automatic sequenator. Results The length of acquisitive HEV-CCC220 was 7193 bp coding 2397 amino acid . The gene of HEV-CCC220 was composed of 5’-UTR(nt 1-9),3’- UTR(nt 7152-7193) and three open reading frames(ORFs). Its homology with HEV IV was obviously higher than that with HEV I-III. The whole genome sequence nucleotide homology was 83.2%-85.0%,85.0%-93.9% and80.6%-86.7% as compared with 300nt and 98nt partial nucleotide sequence of ORF2.The homology comparison of various areas and various lengths of nucleotide was different. Conclusion The HEV-ccc220,which sporadically happened in Changchun,is the subtype IV of HEV.The 300nt of ORF2 can replace the whole gene order to perform genotype analysis of HEV.

Objective To study the effecet of oxidized low-density lipoprotein(Ox-LDL) on endothelium-dependent relaxation and mechanism of susceptibility to atherosclerosis (AS) in hyperlipdemic male patients. Methods LDL isolated from 13 normal patients and 29 hyperlipidemic patients were modified by CuSO₄. The amount of malondialdehyde (MDA) was measured by TBARS. The amount of lysophosphatidylcholine (LPC) was determined by the Bartlett. Endothelium-dependent relaxation was produced by acetylcholine. Results After LDL from normal and hyperlipidemic patients were modified by CuSO₄, the amount of MDA was increased (P<0.01). LPC in Ox-LDL in two groups were higher than those in their respec tive native LDL(N-LDL). N-LDL contained similar levels of LPC in males and females in two groups, but Ox-LDL from males produced more LPC than that from females in both groups (P<0.05), Ox-LDL from males of hyperlipidemic patients contained the highest level of LPC. Ox-LDL from males and females in both groups inhibited endothelium-dependent relaxation. In hyperlipidemicpatients, Ox-LDL from males caused greater impairment of endothelium-dependent ralaxation than that from females, the effect of gender was minimal within the normal group. Correlation analysis ind icated that the degree of impairment of endothelium-dependent relaxation of male and female hyperlipidemic patients was correlated with the production of LPC during oxidation, (r＝0.592，P<0.05；r＝0.816P<0.05). Conclusion The mechanism of susceptibility to AS in hyperlipdemic male patients is related with the increase of LPC production and greater endothelial dysfunction in Ox-LD L.

Objective To study the expressions of metalloproteinase 2(MMP-2) and tissue inhibitors of metalloproteinase 2 (TIMP-2) in lung cancer, and their relationships with microvessel density(MVD). Methods The expressions of MMP-2 ,TIMP-2 and Ⅷ factor were tested by immunohistochemical staining in 90 cases of lung cancer tissue and 40 cases of para-cancer lung tissue. Results The immunohist ochenmical staining results analyzed by IPP software indicated that MMP-2 and TIMP-2 expressed in all pathological types of lung cancer. The expreesion level of MMP-2 in lung cancer tissue was higher than that in para-cancer tissue(P<0.001),it was higher in adenocarcinoma than that in squamous carcinoma(P<0.05),and was the highest in SCLC(P<0.01);the expresion level in lung cancertissue with lymph node metastasis was higher than that in lung cancer tisse without lymph node metastasis(P<0.001).The expression level of TIMP-2 in lung cancer tissue was lower than that in para-cancer tissue(P<0.00 1),there was no significant difference between various pathological types of lung cancer(P<0.05) ;the expression level of TIMP-2 in lung cancer with lymph node metastasis was lower(P<0.05) .The expression level of MMP-2 increased and TIMP-2 decreased with the increasing of MVD.Conclusion Expressions of MMP-2 in differet tissue are on different levels. The expressions of MMP-2 and TIMP-2 protein are closely correlated with the MVD of lung cancer.

Objective To study the expressions of fragile histidine triad (FHIT) in cervical cancer and cervical intraepithelial neoplasia (CIN) ,and the relationship between the FHIT expression and the tumor occurance and development in cervical cancer. Methods The expressions of FHIT in cervical cancer tissue of 35 patients, 39 CIN samples and 16 normal cervix tissue were detected by SPimmunohistochemical method and the relations with occurance, pathologic type, histological grade ,clinical pathologic staging and lymph node metastasis in cervical cancer were analyzed. Results FHIT expressed in normal cervical tissue, CIN and cervical cancer tissue, which showed a decreasing trend (P<0.01). FHIT expression in squamous carcinoma was higher than that in CIN tissue（P<0.01）, it also was higher in squamous carcinoma than that in adenocarcinoma（P<0.05）, and it was higher in lymph node metastatic cases than that in cases without lymph node metastasis. There was no significant difference between histological grade, different grade CIN tissue and clinical pathologic staging (P>0.05). Conclusion Expression of FHIT has close relations with the tumor occurance and development in cervical cancer ,which could provide a reference foundation to monitor lapse of CIN and judge prognosis of cervical cancer.

Objective To investigate the relationship between anti-trophoblast membrane antigens(TA) antibodies and hypertensive disorders complicating pregnancy(HDCP). Methods Enzyme linked immunosorbent assay(ELISA) was used to detect anti-TA IgG and IgM in both maternal and umbilical serum samples of 40 normal pregnant women and 92 HDCP women(23 gestational hypertension or mild preeclampsia, 41 severe preeclampsia and 28 eclampsia). Results The positive rates of anti-TA IgG and IgM in maternal serum samples with HDCP , eclampsia or severe preeclam psia were significantly higher than that in normal pregnant women ，the positiv e rate of anti-TA IgM increased significantly with the aggravation of HDCP(P<0.05). The positive rates of anti-TA IgG in umbilical serum samples with HDCP, eclampsia or severe preeclampsia were significantly higher than that in normal pregnant women(P<0.05). The positive rates of anti-TA IgM in patients with HDCP or eclampsia were significantly higher than that in normal pregnant women(P<0.05).The positive rate of anti-TA IgM in HDCP patients with fetal growth restriction (FGR) was significantly higher than that in those without FGR(P<0.05). Conclusion The anti-TA IgG and IgM in maternal and umbilical serum increase in HDCP patients, and the increasing of anti-TA IgM in maternal serum is associated with the aggravation of HDCP.

Objective To study the effects of　recombinant human erythropoietin (r-HuEPO) on RBC immunity in 　chronic renal failure (CRF) patients. Methods Forty-five CRF patients were divided into two groups : 20 patients without r-HuEPO (control group) and 25 patients with r-HuEPO more than one month (experimental group). The RBC-C_3bRR and RBC-ICRR in two groups were counted under microscope and the correlations between HB or CRE and RBC-C_3bRR were calculated. Results The concentrationd of Hb, RBC-C_3bRR in experimental group were much higher than those in control group(P<0.05). The RBC-ICRR in experimental group was much lower than that in control group(P<0.05). There was a positive correlation between HB and RBC-C_3bRR(r=0.56,P<0.05). There was a negative correlation between CRE and RBC-C_3bRR(r=－0.55,P<0.05). Conclusion r-HuEPO can improve the anaemia in CRF patients,improve the activities of C_3b receptors, improve the RBC immunity and depress the CRE concentration.

Objective To observe the mechanism of the change in runx3 in gastric carcinoma itsdifference and with p53 mutation. Methods A total of 30 cases of gastric carcinoma were taken from clinical operation. PCR-SSCP method was used to detect mutations in exons 2，3，4 of runx3 gene in the gastric carcinoma.The methylation status of runx3 gene was examined by methylation-specificpolymerase chain reaction (MS-PCR), PCR-SSCP method was used to detect mutations in exons 5，6，7，8 of p53 gene in the gastric carcinoma. Results Two cases with mutation were found ,methylation of runx3 promoter region was confirmed in 87% (26/30) specimens of gastric carcinoma, 53.3％（16/30）cases were confirmed with mutati on in the p53 gene. The rate of methylation of runx3 was higher than the rate of mutation of p53（P<0.05）.Conclusion The gene mutation of runx3 is not the change of the molecular biology of gene in the gastriccarcinoma.High frequent methylation in the promoter region of runx3 is primarychange in the gastric carcinoma; compared with the mutation in the p53 gene, high frequent methylation in the promoter region of runx3 has genetic specificity and runx3 is pivotal gene in the development of gastric carcinoma.

Objective To investigate the relationships between Cyclooxygenase (COX-2) and angiogenesis and expressions of COX-2 and vascular endothelial growth factor (VEGF) in thyroid cancer and its clinical significance. Methods Immunohistoc hemistry was used to detect the expressions of COX-2 and VEGF in 60 thyroid cancer,15 thyroid adenomas and 10 normal thyroid tissues. Results The expression rat es of COX-2 and VEGF in thyroid cancer were higher than those in thyroid adenomas and normal tissues(P<0.01).The expressions of COX-2 and VEGF in thyroid cancer presented positive correlation(r＝0.636, P<0.01).The expression rates of COX-2 in thyroid papillary cancer and undifferentiated cancer were 47.1% and 90.0% .The expression rates of VEGF in thyroid papillary cancer and undifferentiated cancer were 71.4% and 100%.The expression rates of COX-2 and VEGF in TNM Ⅰ and Ⅱ were lower than those in TNM Ⅲ and Ⅳ(P<0.05).The expression rates of COX-2 and VEGF in thyroid cancer with lymph node metastasis were higher those that without lymph node metastasis(P<0.05). Conclusion COX-2 and VEGF express highly in thyroid cancer.The expressions of COX-2 and VEGF in thyroid cancer correlates with pathological type,TNM phase and lymph node metastasis of thyroid cancer.

Objective To investigate the expression of hMLH₁ in gastric cancer and its significance, Methods The expressions of hMLH₁ protein and mRNA were dete cted by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) in 68 cases of gastric cancer tissues, 33 cases of para-cancer tissues and 35 cases of normal gastric tissues. Apoptosis was assayed by terminal deoxytransferase mediated dUTP nicked labeling. Results The positive expression rates of hMLH₁ mRNA were 63.3% (43/68), 100% (33/33) and 100% (35/35), and those of protein were 61.8% (42/68), 93.9% (31/33), 100% (35/35), respectively, in gastric cancer tissues, para-cancer tissues and normal gastric tissues. There was a correlation between the expression of hMLH₁ and clinical grade of gastric cancer (r=0.531,P<0.05) . The apoptotic index in the gastric cancer tiss ue with positive expression of hMLH₁ ［(3.652±0.241)%］ was higher than that in the gastric cancer tissue with negative expression of hMLH₁　［(1.887±0.212)%］(P<0.01). Conclusion The low expression of hMLH₁ in gastric cancer tissue indicats that hMLH ₁ expression may play an important role in the pathway of carcinogenesis of gastric cancer.

Objective To compare the bond strength between three different treatment factors. Methods Sixty extracted human premolars were randomly divided into 3 groups(n=20). Edgewise bracket was bonded on each tooth.　The 24 h shear bond strength and tensile bond strength in 3 groups were tested respectively.The adhesive remnant index was recorded. Results The significantly higher shear bond strengths were observed in the polishing group with tental politure and the group that bonded with light-cured fluoride glass ionomer cements after polishing（Group 1∶5.973±1.519, Group 2∶9.017±3.278, Group 3∶7.862±2.437, P<0.05,unit:MP）.Adhe sive remnant index score showed that polishing with tental politure could increa se the bond strength between the enamel surface and adhesive significantly （P<0.05）, especially in the tensile bond strength（P<0.01). The group that bonded with light-cured fluoride glass ionomer cements after polishing had a sign ificantly higher tensile bond strength between the enamel surface and adhesive than no cleaning group（P<0.05）. Conclusion The tental politure and light-cured fluoride glass ionomer cements could be used in the orthodotics bracketbonding .

Objective To study the change curve of ADH in central diabetes insipidus after neurosurgical operation, and analyse the clinical significance of diffe rentiating the types of central diabetes insipidus through the change curve. Methods The serum level of ADH was observed in 158 central diabetes insipidus patients undergone neurosurgical operation. The change curve of ADH in different types of central diabetes insipidus were painted, including transient, continuous and triphasic diabetes insipidus. Results The serum concentrations of ADH in transient and triphasic diabetes insipidus decreased after operation, and arrived the low point (41.7% and 63.6% of the preoperative serum concentration of ADH) after 2 d. The serum concentration of ADH in transient diabetes insipidus recovered to the preoperative level after 10 d. The serum concentration of ADH in triphasic diabetes insipidus arrived the peak after 7 d, then descended. The serum concentration of ADH in continuous diabetes insipidus fell in postoperative 1 d, and arrived the low point (33.3% of the preoperative serum level of ADH) after 7 d. The serum concentration of ADH in continuous diabetes insipidus postoperative 2 weeks was lower than preoperation. Conclusion Though the concentration change curve of ADH, the transient diabetes insipidus can be distinguished from continuous and triphasic diabetes insipidus. The correct differential diagnosis and seemly treatment are beneficial for patients’ prognosis.

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