To observe the effects of taurolidine on radiosensitivity of B16-F10 cells of murine melanoma via the enhancement of Bax and Bad proteins and induction of Bcl-2 protein. Methods The apoptosis of B16-F10 cells was assessed after treated with 0,10,25,50,100 and 150 μmol•L^-1 taurolidine,clone survival assay was used to detect the radiosensitivity of B16-F10 cells,and protein expressions were determined by Western blotting.Results The apoptosis of 5% cells was induced in a dose-and time-dependent manner after B16-F10 cells were treated with 50 μmol•L^-1 taurolidine.The survival rate decreased after treated with taurolidine in combination with 2 Gy X-irradiation with the increase of taurolidine concentration and doses of irradiation（P<0.05）,and the ratio of radiosensitivity (SER D₀ and SER D_q) also increased with the increase of its concentration,there was significant difference between 50 μmol•L^-1 taurolidine group and 10 μmol•L^-1 taurolidine group（P<0.05）; meantime, the level of proapototic protein Bax and Bad increased and the level of antiapoptotic protein Bcl-2 reduced. Conclusion Taurolidine in combination with irradiation can enhance the radiaosensitivity by the mediation of Bcl-2 family protein.

To Study the effect of Gross Saponins of Tribulus Terrestris on active potential and potassium current of ischemic cardiocytes in guinea pigs， and approach its initial anti-arrhythmia effect and mechanism.Methods The cardiocytes of adult guinea pigs were isolated and hypoxia models were set up,the effects of injected GSTT on the cellular membrane active potential and electricity flow of potassium channel were recorded with whole cell patchclamp system. Results Compared with model group,the active plateau period(50% and 90%) of myocytes in GSTT 30 mg•L^-1 group prolonged(P<0.01).When the electric potential at －40 mV，the test electric potential was from －100 mV to +100 mV，step order at +10 mV，and duration at 300 ms.The myocyte potassium ion in control group closed in this condition.The peak currents in GSTT 10 and 30 mg•L^-1 groups were （1.19±0.20）and（1.15±0.1）nA，,and the difference was significant compared with model group（1.50 nA±0.20 nA］ (P<0.05 and P<0.01). Conclusion GSTT could extend action potential duration,prevent potassium current,decrease the cellular membrane electricity alternations so that it could protect the cardiac myocytes from hypoxia damage.

To investigate the effect of urotensin Ⅱ (UⅡ) on proliferation of the rat mesangial cells (MC) cultivated with high glucose culture medium in vitro and explore its mechanism and significances. Methods The rat kidney MC cultivated with high glucose in vitro were divided into five groups: control group and different concentrations of UⅡ(10^-7,10^-8,10^-9,10^-10mol•L^-1)groups,incubated at 37℃ for 24 h,the proliferation rates in various groups were determined by MTT assay，the ratios of MC in S phase in various groups were detected by flow cytometry，the BrdU contents in various groups were determined by BrdU incorporation method.Results In MTT experiment,the MC proliferation rates in UⅡ10^-9 and UⅡ10^-8 mol•L^-1 groups were significantly higher than that in control group (P<0.05),and the results in UⅡ 10^-10 and UⅡ 10^-7mol•L^-1 groups had no significant change compared with control group (P> 0.05); In flow cytometry experiment,the ratios of MC in S phase in UⅡ 10^-9 and UⅡ 10^-8 mol•L^-1 groups were increased significantly compared with control group (P< 0.05),and the results in UⅡ 10^-10 and UⅡ 10^-7 mol•L^-1 groups had no significant change compared with control group (P> 0.05); In BrdU-ELISA test ，BrdU contents in UⅡ 10^-9 and UⅡ 10^-8 mol•L^-1 groups were significantly lower than that in control group (P< 0.05),while the results in UⅡ 10^-10 and UⅡ 10^-7 mol•L^-1 groups had no significant change compared with control group (P> 0.05). Conclusion UⅡ at the concentrations of 10^-9 and 10^-8 mol•L^-1 have obvious role in promoting proliferation of rat MC.

Abstract:Objective To explore the effects of angiotensinⅡ products on urotensinⅡ(UⅡ) and its receptor GPR14 expression in cultured renal interstitial fibroblasts and the intervention of ginkgo bioba extract(GBE). Methods The rat renal fibroblasts were cultivated in vitro,untreated cells were acted as normal control group,AngⅡ as the stimulating factor,the cells were treated with GBE with concentrations of 0,25,50,100 and 200 g•L^-1,the mRNA and protein levels of UⅡ and GPR14 were measured by RT-PCR and Western blotting,the content of collagenⅠ in supernatant was detected by ELISA method. Results Compared with AngⅡ group,the mRNA and protein levels of UⅡ and GPR14 were significantly down-regulated (P<0.01 or P<0.05),the contents of ColⅠ in supernatant were markedly decreasedin AngⅡ+GBE group(P<0.01 or P<0.05). Conclusion GBE　 can down regulate the UⅡ and GPR14 mRNA expression,decrease the protein levels of UⅡ and GPR14,and reduce the ColⅠ secretion in cultured fibroblasts.

To investigate the effect of midazolam on the synaptic long-term potentiation (LTP) in the CA₁ region of rat hippocampal slices，and elucidate the mechanisms underlying the effect of midazolam on memory. Methods Hippocampal slices (400 μm thick) were obtained from the brains of 126 male Sprague-Dawley rats that were decapitated.The slices were incubated in artificial cerebrospinal fluid (ACSF) at room temperature for at least 120 min before use.Forty-two slices were randomly divided into 6 groups (n=7): control group， midazolam groups with doses of 0.1,0.5,1.0,2.0 and 5.0 μmol•L^-1. All the slices in each group were perfused with ACSF,midazolam 0.1,0.5,1.0,2.0 or 5.0 μmol•L^-1,respectively.Extracellular microelectrode recording technique was performed to record the evoked population spikes (PS) in CA₁ region.Another eighty-four slices were randomly divided into 12 groups (n=7):LTP group,midazolam-LTP groups with doses of 0.1,0.5,1.0,2.0,5.0 μmol•L^-1,picrotoxin group,bicuculline group,CGP35348 group,picrotoxin + midazolam group,bicuculline + midazolam group and CGP35348 + midazolam group.All the slices in each group were perfused with ACSF and the drugs that were applied in according groups.PSs were recorded for at least 30 min before LTP in each group.For LTP induction,high-frequency stimulation (HFS) conditioning pulses (100 Hz/s) were applied to the Schaffer collateral-commissural pathway of hippocampus using bipolar stimulating electrode. The changes in PS amplitude after HFS were analyzed in each group.Results The PS amplitude of the rat hippocampal slices in midazolam 1.0,2.0 and 5.0 μmol•L^-1 groups were significantly decreased compared with control group (P<0.05 or P<0.01).The PS amplitude in LTP group after HFS was significantly increased by 52%±12% compared with pre-HFS(P<0.01).The PS amplitude was not significantly changed after HFS in midazolam-LTP 0.1 μmol•L^-1　group,when compared with LTP group(P>0.05). Compared with LTP group,the PS amplitudes in midazolam-LTP 0.5,1.0,2.0 and 5.0 μmol•L^-1 groups after HFS were decreased significantly (P<0.01).The PS amplitudes after HFS in picrotoxin + midazolam group and bicuculline + midazolam group were dramatically increased compared with pre-HSF and midazolam-LTP 2.0 μmol•L^-1 group (P<0.01),but there was no difference compared with LTP group(P>0.05).The PS amplitude after HFS in CGP35348 + midazolam group had no significant difference compared with pre-HSF and midazolam-LTP 2.0 μmol•L^-1 group(P>0.05),but it was significantly lower than that in LTP group (P<0.01). Conclusion The inhibition of LTP induction in hippocampus of rats may contribute to midazolam-induced deficits in memory,and the underlying mechanism is involved in the activation of γ-aminobutyric acid (GABA)A receptor in hippocampus.

To transfect PCDNA3.0-HIF1-α into the endothelial progenitor cells(EPCs),in order to provide the foundation for exploring myocardial vascular repairing capacity and angiogenesis of the EPCs transfected with HIF gene. Methods The EPCs were separated by the method of density gradient separation in the human bone marrow,and then the cells were inoculated in the fibronectin-packaged petri dishes complete medium with the concentration of 1×10⁶•mL^-1,and cultivated for 14 d. The expressions of the endothelial cell-specific components ecNOS and flk-1 were detected with RT-PCR; the cells were stained with acLDL-Dil and FITC-UEA-1; the PCDNA3.0-HIF1-α plasmid was transfected into the EPCs mediated by Lipofectamine^TM 2000,the transcription of HIF1-α was determined with RT-PCR; the expression of VEGF was detected with enzyme-linked immunosorbent assay (ELISA); the expression of HIF1-α was detected with Western blotting. Results After cultivated for 5 d,some round monocytes transformated into spindled inherent EPCs;7 d later,there were some cell colonies containing dozens of cells;10 d later,there were obvious cell clones; RT-PCR showed ecNOS band in 548 bp,flk-1 band in 819 bp; acLDL-Dil and FITC-UEA-1 double-staining was positive; RT-PCR showed specific band in 383 bp; ELISA showed the content of VEGF in the supernatant of EP Cs transfected with HIF1-α was （20.53±2.33） μg•L^-1,and it was （3.96±1.67） μg•L^-1 in the EPCs transfected without HIF1-α,there was significant difference between two groups (P<0.05).Western blotting showed specific band in 120 000. Conclusion PCDNA3.0- HIF1-α is successfully transfected into EPCs.

To determine the effect of heroin on the gene expression of proliferating cell nuclear antigen(PCNA) in rat C6 glioma cells，and investigate the molecular mechanism of heroin addiction.Methods The rat C6 glioma cells were treated with heroin with doses of 0－100 mg•L^-1 for 24 h,MTT assay was used to detect the viability of cells,the gene expression level of PCNA was determined by RT-PCR. Results When the C6 cells were treated with heroin with doses of 10,20 and 100 mg•L^-1,the cell viabilities were 90.62%，80.97%,and 62.73%,there were significant differences compared with control group(P<0.05).When treated with 20 mg•L^-1 heroin for 48 h,the expression level of PCNA mRNA of C6 glioma cells was decreased as compared with corresponding control cells,the ratio was 0.297（P<0.01）.Conclusion Heroin with dose of 20 mg•L^-1 can inhibit the growth of C6 glioma cells by 30 percent.It may be related to heroin addiction.

To investigate the effects of stress associated with nicotine on experimental periodontal breakdown of ligature-induced periodontitis in rats. Methods Forty male Wistar rats with the neck of left maxillary second molar ligated by nylon were used.The animals were randomly divided into four groups: Ni group,0.7 mg of nicotine•kg^-1•d^-1 (intraperitoneal); S group,stress (immobilization-12 h•d^-1); Ni+S group,stress (immobilization-12 h•d^-1) associated with an intraperitoneal injection of 0.7 mg of nicotine•kg^-1•d^-1; C group：saline solution. Two rats of each group were sacrificed at the day of 2,4,6,8,10 separately and the furcation areas of the second maxillary molars were examined histologically and histometrically. Results Intergroup analysis revealed a higher bone loss rate in the animals of Ni+S group compared with the other three groups,the absorption rate of alveolar bone at the day of 6,8,10 in experimental groups were higher than that in C group (P<0.01); the absorption rate of alveolar bone at the day of 6,10 in Ni group and Ni+S group were higher than that in S group (P<0.01) and at the day of 10,it was higher in Ni+S group compared with Ni group (P<0.05); and there were significant differences at the day of 6,8,10 between Ni group and C group (P＜0.05) and at the day of 6 between Ni group and S group (P<0.01). Conclusion Stress can significantly enhance the destructive effects of nicotine on the periodontal tissue,and they have synergetic effect in the destructive process of periodontal tissue.

To study the induction of amelogenin in formation of hard tissues in the pulp tissues,to provide experimental foundation for research in formation of hard tissues induced with amelogenin. Methods Sixteen male Wistar rats were divided into control group and experimental group.Cells and matrix structures and amelogenin expression in the growing ends of cyclophosphamide(CP)-affected incisors were examined by histological and immunohistochemical methods.Results Experiment group: extensive edematous changes were noted in the apical pulp tissue of the incisor associated with disappearance of odontoblasts facing to presecretory and secretory ameloblasts. The cell layer of young ameloblasts was therefore lined directly with pulp tissue in which some osteodentin-like tissue was present near the ameloblasts. Immunoreactions for amelogenins were observed in the labial side of the pulp,particularly around the newly induced hard tissues in the affected region. Conclusion Amelogenins has induction in formation of hard tissues in dental pulp following the inhibition of formation of odontoblasts by CP.

To study the effect of over-expressing exogenous and RNA-interfered endogenous NDRG3 on colony-formation of prostatic carcinoma cells. Methods NDRG3 expression plasmids were stably transfected to PC3 prostatic carcinoma cell lines.The NDRG3-stable transfected PC3 sublines were studied along with parental and empty vector transfected PC3 cells as controls; Endogenous NDRG3 was silenced by RNA interference technique and appreciated using Western blotting technique.Colony-formation experiments were performed in control group,GFP-siRNA group and NDRG3-siRNA group. Results Over- expressing exogenous NDRG3 of PC3 formed notably more colonies than the parental and empty vector transfected PC3 cell lines.In addition,the total colonies and big colonies in control group and GFP-siRNA group were more than that in NDRG3-siRNA group in RNA interference test（P<0.05）. Conclusion NDRG3 can enhance carcinogenic potential and colony-formation viability of prostatic carcinoma cells.

To find the molecular mechanism of apoptosis of multi-drug resistance cells lines(MDR-MG-63) induced by semiconductor laser and provide theoretical basis for its clinical application. Methods MDR-MG-63 cells were divided into five groups:control group and semiconductor laser groups(30,60,90,120J•cm^-2).The cell viability was detected by MTT assay; reactive oxygen species (ROS) was detected by flow cytometry;the protein expressions of caspase-8,caspase-9 and caspase-3 were assayed by Western blotting. Results Compared with control group,the cell viabilities decreased gradually with the increasing of dose in semiconductor laser groups (60,90,120J•cm^-2)(P<0.05 or P<0.01),a lot of ROS were found (P<0.05) after multi-drug resistance MG-63 cells lines were irradiated,at the same time the expression levels of caspase-9 protein and caspase-3 protein were increased(P<0.05 or P<0.01) in a dose-dependent manner,but the level of caspase-8 protein didn’[KG-*5]t obviously change （P>0.05）;the differences of the cell viability,ROS,the expression levels of caspase-9 protein and caspase-3 protein between semiconductor laser(30J•cm^-2)group and control group were not significant (P>0.05).Conclusion Semiconductor laser can induce apoptosis of MG-63 cells by mitochondrion pathway,and the process may be associated with the increasing of ROS.

To observe the effect of methylseleninic acid (MSA) on proliferation and apoptosis of the prostatic cancer cells DU145 and approach the mechanism.Methods After DU145 cells were treated with 1.25,2.50 and 5.00 μmol•L^-1MSA,the cell viability was examined by MTT test. The techniques of acridine orange staining and flow cytometry were used to analyze apoptotic cycle and calculate the apoptotic index. The expression of active caspase-3 was detected with immunocytochemistry. Results After DU145 cells were treated with 1.25,2.50 and 5.00 μmol•L^-1MSA,the growth inhibitory rates of cell line were 16.35%,38.10% and 73.54 %,respectively,the inhibitory rate increased with the increasing of MSA concentration in a dose-dependent manner. There existed obvious differences between three groups（P<0.01）.Acridine orange staining results showed that the cells were apoptotic state when treated with 1.25,2.50 and 5.00 μmol•L^-1MSA,the number of apoptotic cells increased with MSA dose increasing. Flow cytometry results showed that when the cells were treated with 1.25,2.50 and 5.00 μmol•L^-1MSA for 48 h,the apoptotic rates were 5.34%,8.71% and 13.6%, respectively; there existed obvious differences between three groups （P<0.05）.The expression of active caspase-3 was upregulated with MSA dose increasing.Conclusion MSA can induce the apoptosis of prostatic cancer cells DU145 and has significant anti-tumor effect,its mechanism may be related to activate the pathway of caspase.

To set up the method to isolate and purify rat bone marrow mesenchymal stem cells(MSCs) and to investigate the inducing effect of injured liver tissue homogenate.Methods MSCs were isolated and purified from bone marrow of young rat by density gradient centrifugation and adherent culture.MSCs were induced in L-DMEM with 10% injured liver tissue homogenate and 10%FBS in vitro.The expressions of alpha-fetoprotein(AFP) and albumin(Alb) were detected by immunohistochemistry at different time.Results MSCs became epithelia-shaped cells after cultivated in L-DMEM with 10% injured liver tissue homogenate and 10%FBS in vitro. Most cells were AFP positive after cultivated for 7 d(75.2%),compared with control group there was significant difference( P<0.05); AFP positive rates were respectively 35.2%,12.3% and 2.0% after cultivated for 14,21 and 28 d,there were significant differences between various groups(P<0.05); Only a small number of cells were Alb positive after cultivated for 7 d(14.6%).Alb positive rates were respectively 67.6%,88.9% and 95.6% after cultivated for 14,21 and 28 d,compared with control group and between different time points there were significant differences (P<0.05). Conclusion A method for isolation and purification of MSCs from bone marrow in vitro has been established.Injured liver tissue homogenate could induce the ex pressions of AFP and Alb in MSCs.

To explore the protective effect of pulmonary artery perfusion with lung-protective solution and oxygenated blood on lung morphous and function of dogs during cardiopulmonary bypass (CPB).Methods Eighteen adult mongrel dogs were randomly divided into three groups.During CPB,the lungs of the dogs in first group were perfused with diluted oxygenated blood from CPB-machine(group B),the lungs of the dogs in second group were perfused with cold lung-protective solution (group P),the lungs of the dogs in third group were perfused with nothing (control group).Malondialdehyde (MDA) of lung tissue,alveolar-arterial oxygen gradient (P_［A-a］O₂),the ratio of white blood cells in BALf and lung function were measured at different time points （before CPB and 15 and 60 min after CPB）,lung biopsies were also made. Results The levels of airway resistance at the 15th and 60th minutes after CPB in group B were higher than before CPB (P<0.05). The levels of airway resistance at the 15th and 60th minutes after CPB in group B were lower than those in control group at the same time points (P<0.01).The levels of pulmonary vascular resistance at the 15th and 60th minutes after CPB in each group were lower than before CPB (P<0.01).The levels at the 15th and 60th minutes after CPB in group B and control group were higher than the levels at the same time points in group P(P<0.05).The levels of PaO₂/FiO₂ at the 15th and 60th minutes after CPB in each group were higher than before CPB (P<0.01).The levels of PaO₂/FiO₂ at the 15th and 60th minutes after CPB in group B and control group were lower than the levels at the same time points in group P(P<0.05).The level of P_［A-a］O₂ at the 60th minute after CPB in group P was lower than those in group B and control group (P<0.001).The level of MDA and neutrophil percentage in BALF at the 90th minute after CPB in group P were lower than those in group B(P<0.05) and control group (P<0.001,P<0.05),and the level s in B group were lower than those in control group (P<0.001,P<0.05).Morphological analysis revealed that the percentage of pathological lesion and alveolar hemorrhage grade values in group P were than those in group B(P<0.05) and control group (P<0.001),and they were lower in group B than control group (P<0.001) Conclusion The morphous and function damage occurs in lungs after CPB.Both oxygenated blood and lung-protective solution have protective effect on lung morphous and function.

To investigate the mechanism of apoptosis of lung carcinoma cell line NCI-H460 induced by 20(S)-ginsenoside Rg3 (SPG-Rg3).Methods MTT assay was used to detect the proliferation rate of NCI-H460 cells.The cells were treated with SPG-Rg3 and divided into three groups based on IC₅₀ value: SPG-Rg3 25 mg•L^-1 group，SPG-Rg3 50 mg•L^-1 group and SPG-Rg3 100 mg•L^-1 group.Control group was also set up.Then the AO/EB fluorescence double-dye technique,immunocytochemistry staining,Fluo-3 staining and Rhodamine 123 uptake assay approaches were adopted to observe the apoptotic rate,expression levels of Bcl-2 and Bax proteins,concentration of cytoplasmic free Ca²⁺ and mitochondrial transmembrane potential of NCI-H460 cells. Results SPG-Rg3 obviously inhibited the cell proliferation,showing a positive correlation between inhibitory effect and concentration (r=0.764,P<0.05)，and the IC₅₀ was 47.97 mg•L^-1.There were visible apoptotic changes observed in SPG-Rg3 groups.The protein expression level of Bcl-2 in SPG-Rg3 100 mg•L^-1 group was less than that in control group(P<0.01).The protein expression levels of Bax in SPG-Rg3 groups were more than that in control group（P<0.001）,and they were positively correlated with the drug dose（P<0.01）.The ratios of Bax to Bcl-2 in SPG-Rg3 groups were bigger than that in control group, while the mitochondrial transmembrane potential showed an opposite situation(P<0.01) and it decreased when the drug dose increased.There were significantly differences between various groups (P<0.01).The concentrations of cytoplasmic free Ca²⁺ in SPG-Rg3 groups were higher than that in control group(P<0.01). Conclusion SPG-Rg3 could obviously inhibit the cell proliferation and induce the apoptosis.The mechanism is concerned with that SPG-Rg3 could up-regulate the expression level of Bax protein and increase the ratio of Bax to Bcl-2,down-regulate the mitochondrial transmembrane potential and increase the cytoplasmic free Ca²⁺ concentration,lead to the induction of apoptosis via mitochondrial pathway.

To study the inhibitory effects of dehydroepiandrosterone (DHEA) and its metabolites-dehydroepiandrosterone sulfate (DHEAs) on the proliferation of HepG2 and HT-29 and their mechanism.Methods HepG2 and HT-29 were incubated by DHEA or DHEAs with different concentrations (1,10,50,100 and 200 μmol•L^-1) for 8,24,48,72 h and routine culture was used as control.The inhibitory rate was detected by using MTT chromometry and BrdU assay respectively.The activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR),glucose -6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were examined simultaneously. Results ①MTT chromometry：DHEA with different concentrations obviously inhibited the growth of HepG2 and HT-29 cells compared with control group（P<0.05）.With the concentration of 100 μmol•L^-1 and incubated for 24 h,the inhibitory effect of DHEA decreased markedly（P<0.05）,whereas there was no significant difference of inhibitory effect between DHEAs group and control group （P>0.05）.②BrdU assay：the growth of cells were significantly inhibited by DHEA with concentrations of 50,100 and 200 μmol•L^-1,especially to HepG2 cells（P<0.05).③To HepG2 cells,HMGR activity could be inhibited by DHEA,and its inhibitory rate was 62%,while the inhibitory rate on HT-29 cells was 51%,however,DHEAs had no significant effect.④The inhibitory rate on G6PD activity was 85% by DHEA with the concentration of 100 μmol•L^-1,whereas the inhibitory rate by DHEAs was about 6%.⑤DHEA and DHEAs had no significant effect on LDH activity（P>0.05）. Conclusion DHEA has strong anti-proliferative effects on both HepG2 and HT-29 cell lines and inhibitory effects on the activities of G6PD or HMGR,however,DHEAs has no obvious effect.

To establish the method to express leukemia inhibitory factor(LIF) in fetal fibroblasts in order to provide theoretical foundation for establishment of transgenic animal model.Methods Using the fetal of 13.5 d ICR mouse, the primary fetal fibroblasts were cultivated by trypsin enzyme digestion.The lineared eukaryotic expression vector pcDNA3.1-LIF was transferred to the fetal fibroblasts of mouse with liposome induction.The positive cells were selected by G418,genome DNA of the positive cells was extracted and sequenced. Results The primary fetal fibroblasts of mouse were successfully obtained by isolating and culturing, and fetal fibroblasts expressing LIF were established by transferring.The sequencing result demonstrated that the homology of clone plasmid of positive cells was about 99%.Conclusion Eukaryotic expression vector pcDNA3.1-LIF is successfully transferred to the fetal fibroblasts of mouse.

To study the influence of dihydromyricetin on the oxidation and non-enzymatic glycosylation in impaired glucose tolerance rats.Methods Animal model of impaired glucose tolerance rat was built by intragastrical(i.g.) injection of D-galactose generally.After 8 weeks intragastrical(i.g.) injection of dihydromyricetin(experimental group) and metformin (positive control),the levels of fasting blood glucose and tow-hour postprandial blood glucose,the insulin level and its resistance index,the levels of glycosylated hemoglobin (GHb) and advanced glycation end-products(AGEs) were detected,and the activities of superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px),the content of malondiadehyde(MDA) were determined respectively. Results Compared with model group，dihydromyricetin decreased the level of two-hour postprandial blood glucose (P<0.05),lowered serum insulin level, its resistance index (P<0.05) and the contents of MDA and AGEs in serum (P<0.05) . In addition,it increased the activity of GSH-Px in serum (P<0.05); but there was no difference of both the content of GHb and the activity of SOD between all groups(P>0.05).Conclusion Dihydromyricetin can significantly improve the state of impaired glucose tolerance rats; the mechanism may be related to its effect of improving the antioxidant activity of GSH-Px,inhibiting non-enzymatic glycation,lowering body oxidative stress and insulin resistance and promoting the use of blood glucose in peripheral organizations.

To evaluate the effect of basic aloe mastic on the healing process after tooth extraction in rats. Methods The models of tooth extraction wound were established by extracting the left and right maxillary first molares in 60 Wistar rats, and randomly divided into two groups. Right sides of teeth extraction socket were used as experimental groups，experimental group 1 was filled with 30% aloe mastic , experimental group 2 was filled with 50% aloe mastic. Left sides of teeth extraction socket were used as control group. The histological observation was performed after tooth extraction at 2,4,6,8,11,and 15 days. Results There was a significant difference of the wound areas between experimental groups and control group at early stage (15 d) after tooth extraction (P<0.05). The histological observation confirmed that in experimental groups, the new bone formation was earlier and quicker than that in control group. Conclusion Aloe mastic has the effect to promot the proliferation of fibroblasts and the growth of granulation tissue and epithelial cells, it is in favour of the formation of new alveolar bone effectively during wound healing of extraction at early stage. Meanwhile the effect of 30% aloe mastic is better than 50% aloe mastic.

To investigate the effect of phenylacetate(PA) on apoptosis in rat C6 glioma cells and explore the possible mechanism.Methods C6 cells cultivated in vitro were induced by PA,then the apoptosis was detected by TUNEL assay and the expressions of bcl-2,bax,and caspase-3 were determined by immunocytochemistry.Results After treated with PA at concentrations of 0,2.5,and 5.0 mmol•L^-1 for 24 h,the apoptotic rates(%) were 2.5±0.2,10.3±0.5,and 20.6±1.1,respectively; while for 72 h,the apoptotic rates (%) were 2.7±0.3,24.9±0.7,and 42.0±1.7,respectively. The apoptotic rate was remarkably increased with the increasing of PA concentration and treatment time.There were significant differences of apoptotic rates of C6 cells after treated with different conentrations and for different time between experimental and control groups(P<0.05).In C6 cells,the expression levels of bcl-2,bax and caspase-3 were 0.275±0.022,0.214±0.019,and 0.149±0.004,respectively; while treated with PA(5.0 mmol•L^-1) for 72 h,the expression levels of bcl-2,bax,and caspase-3 were 0.244±0.024,0.399±0.030,and 0.206±0.033,respectively. The expression levels of bax and caspase-3 were enhanced by PA (P<0.01)，whereas the expression level of bcl-2 didn’t change (P>0.05).Conclusion PA can induce apoptosis in rat C6 glioma cells,which shows a time- and dose-dependent manner.The mechanism of PA on apoptosis might be related to the up-regulation of bax and caspase-3 expression.

To investigate the effect of N-acetylcysteine(NAC) on matrix metalloproteinases(MMPs) levels in rabbit carotid atherosclerotic plaque and morphological changes. Methods The model of rabbit atherosclerosis was built by injuring internal carotid through arterial canal balloon in vitro.These rabbits were treated with NAC for 6 weeks. Then the injuried arteries stained with HE were observed under light microscope and the levels of MMP-2 and MMP-9 in veinous blood from rabbits were detected by SDS-Page-Zymograph method.Moreover,the number of foam(spume) cells contained MMPs and tissue inhibitor of MMP-2(TIMP-2) were detected by method of immunochemistry.In addition,the status of vascular hyperplasia was proved by the area of new vascular inner membrane,the transverse section of vascular cavity and the thickness of vascular wall.Results The arterial lumen injury neointimal area in NAC group（1.72 mm²±0.26 mm²）was significantly smaller than that in control group（2.66 mm²±0.19 mm²）(P<0.05).The endothelial thickness in NAC group（0.132 mm±0.031 mm）was decreased compared with control group （0.231 mm±0.022 mm）（P<0.05）,and vascular cavity transverse section in NAC group（0.54 mm²±0.15 mm²）was larger than that in control group（0.31 mm²±0.12 mm²）（P<0.05）. On the same lengthof vascular wall,the number of foam cells containing MMPs in NAC group (8.0±1.8) was less than that in control group (22.0±2.2)（P<0.05）.The number of foam cells containing TIMP-2 in NAC group (10.0±1.2)was more than that in control group(6.0±1.6)（P<0.05）. The levels of MMP-2,pro-MMP-2 and pro-MMP-9 in NAC group were remarkably lower than those in control group（P<0.05）. Conclusion NAC has functions of inhibiting the increase of foam cells,preventing vascular sclerosis,inhibiting the degradation of atherosclerosis plaque induced by MMPs,thereby steadying the plaque.

To investigate the genetic association between KPNB3 gene in chromosome 13q32 and schizophrenic phenotypes in Han population from North China. Methods Genomic DNA was isolated from the whole blood samples.A single nucleotide polymorphism,rs626716 (Pst I site) present in the KPNB3 gene,was detected using PCR-based restriction fragment length polymorphism analysis among 752 Chinese Han patients with schizophrenia (case group) and 909 ethnicity-matched healthy controls (control group).The Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness-of-fit χ² test.The frequencies of alleles and genotypes of KPNB3 gene and the relationship between allelic frequencies and the clinical phenotypes of schizophrenia were statistically computed. Results The goodness-of-fit χ² test showed that the genotypic distributions of rs626716 were not deviated from Hardy-Weinberg equilibrium in both case group and control group (P>0.05).The χ² test did not show allelic association and genotypic association for rs626716 (P>0.05).There were no correlations between allelic or genotypic frequencies and the clinical phenotypes of schizophrenia (P>0.05). Conclusion rs626716 locus may not be associated with schizophrenic phenotypes,but the correlation between the other SNPs locus at KPNB3 gene and schizophrenia could not be ruled out.

Objective By researching the effects of G-coupled protein on the expressions of nuclear factor kappa B(NF-κB) and tumor necrosis factor-α(TNF-α) in peripheral blood monocytes induced by fractalkine(FKN) to explore one of possible signal conductive mechanisms of FKN/CX3CR1 and the invention of captopril.Methods Peripheral blood monocytes were isolated from fresh blood of healthy volunteers by Ficoll-Paque gradient centrifugation and divided into control group,FKN group,G-protein inhibitor pertussis toxin(PTX)group,PKC inhibitor RO31-8220 group,inhibitor NF-κB PDTC group,captopril group;the expressions of NF-κB in monocytes from each group were measured by immunohistochemsitry; the supernatant of monocytes was collected from each group,the expressions of TNF-α were dete rmined by enzyme-linked immunosorbent assay(ELISA).Results The expressions of NF-κB and TNF-α in FKN group were increased compared with control group(P<0.05);the expressions of NF-κB and TNF-α in G-protein inhibitor PTX group were decreased compared with FKN group(P<0.05);the expressions of NF-κB and TNF-α in PKC inhibitor RO31-8220 group were decreased compared with FKN group(P<0.05);the expression of TNF-α in NF-κB inhibitor PDTC group was decreased compared with FKN group(P<0.05);the expressions of NF-κB and TNF-α in captopril group were decreased compared with FKN group(P<0.05).Conclusion FKN-CX3CR1 increases the expressions of NF-κB and TNF-α in peripheral blood monocytes;Captopril participates in inhibiting inflammatory and preventing arteriosclerosis perhaps by reducing the expressions of NF-κB and TNF-α in peripheral blood monocytes induced by FKN-CX3CR1;one of signal conductive mechanisms of FKN-CX3CR1 contributing to the progression of atherosclerosis is the following series of cascaded reaction:FKN-CX3CR1→G-coupled protein→PKC→NF-κB→TNF-α.

Objective To investigate the expression of glycosyl-phosphatidyl inositol(GPI) anchored protein on peripheral blood cells from patients with paroxysmal nocturnal hemoglobinuria (PNH) and provide a basis for diagnosis and differential diagnosis of PNH.Methods The percentages of CD55,CD59,CD16 and CD14 expression on the peripheral blood cells from 32 patients with PNH were determined by flow cytometry. Results The　 expression levels of CD55 and CD59 were decreased significantly on the erythrocytes and granulocytes from patients with PNH compared with control group (P<0.001),the expression levels of CD55 and CD59 on granulocytes were lower than those on erythrocytes (P<0.05).The expression levels of CD55 and CD59 on the thrombocytes of PNH patients were lower than those in control group（P<0.001）.The expression levels of CD55 and CD59 on the lymphocytes from 27 PNH patients were lower than those on granulocytes and erythrocytes,the deficiency rate was 30%.The percentage of CD16 expression varied greatly from 34% to 83% on the granulocytes,there was significant difference compared with control group (P<0.001). The deficiency of CD14 clones was detected on the monocytes from PNH patients. Conclusion The diagnosis reliability of patients with PNH is increased by the assay of CD55 and CD59 on all kinds of cells of peripheral blood.The detection of CD16 and CD14 is also an optimal and helpful index to diagnose PNH.

To evaluate the remineralization effect of casein phosphopeptide amorphous calcium phosphate(CPP-ACP) and fluoride complex on artificial enamel subsurface white spot lesions in vitro in order to provide a new method to treat the postorthodontic enamel demineralization.Methods Extracted premolar teeth for orthodontic reason were immersed into lactic acid gel to prepare artificial white spot lesions.Then the specimens were randomly assigned to seven groups: 5.0%CPP-ACFP group,1.0% CPP-ACP group,0.1% CPP-ACPgroup,calcium phosphate saturated solution group,calcium phosphate saturated solution plus fluorid group,deionized water group.Lesion depths and mineral loss were quantitatively determined by microradiography in various groups.Results The lesion depths and mine ral loss after remineralization in each group were significantly reduced(P<0.05) except deionized water group; the lesion depth and mineral loss in 5.0%CPP-ACFP group were significantly lower than those in the other groups(P<0.05,P<0.01); the lesion depth and mineral loss in 5.0%CPP-ACP group were markedly reduced than those in 0.1%CPP-ACP group(P<0.05,P<0.01) .There were no significant differences between 0.1% CPP-ACP group,calcium phosphate saturated solution group and calcium phosphate saturated solution plus fluorid group(P>0.05),but the lesion depths and mineral loss in these three groups were significantly lower than those in deionized water group(P<0.05,P<0.01).Conclusion CPP-ACP has the ability of promoting remineralization of enamel artificial white spot lesions which can be enhanced by fluorid.The remineralization ability depends on the concentration of calcium phosphate stabilized by CPP-ACP.

To study the expression of phosphatidylinositol 3-kinase(PI3K) in ovarian cancer tissue and the role of PI3K in the occurrence and development of ovarian cancer.Methods RT-PCR method was used to investigate the expressions of PI3K mRNA in tissue samples of 10 cases of ovarian cancer,10 benign ovarian tumor and 10 normal ovary.The expressions of PI3K protein in 60 cases of ovarian cancer,24 ovarian benign tumor and 24 normol ovarian tissues were detected by immunohistochemistry. Results The level of PI3K mRNA expression in ovarian cancer tissues was significantly higher than those in normal ovarian tissues and benign ovarian tumor tissues（P<0.01）,and there was no difference between benign ovarian tumor tissues and normal ovarian tissues（P>0.05）.The expression of PI3K protein was located in cytoplasm and (or) nucleus.The positive expression rate of PI3K protein in ovarian cancer was significantly higher than those in normal ovary and benign ovarian tumor (P<0.01); there was correlation between the expression intensity of PI3K protein and pathological grading and clinical stages,the worse differentiation of ovarian cancer,the stronger positive expression of PI3K protein（P<0.05）;the higher clinical stage of ovary cancer,the stronger positive expression of PI3K protein（P<0.05）. Conclusion The expression levels of PI3K mRNA and protein in ovarian cancer tissues are significantly increased and PI3K has the role in the occurrence and development of ovarian cancer.

To investigate the expression of hepatocyte growth factor receptor(c-met) in cervical cancer,and analyze its relationship with proliferating cell nuclear antigen (PCNA). Methods Immunohistochemistry (S-P method) was adopted to detect c-met and PCNA protein expressions in 14 cases of chronic cervicitis and 50 cases of cervical cancer. Results In chronic cervicitis,the positive rate of c-met was 14.3%(2/14),but in all of cervical cancer was 62.0%(31/50), the difference of c-met expression was significant between chronic cervicitis and cervical cancer (P<0.05).The experession rete of c-met in low differentiation group was higher than that in high differentiation group(P<0.05).The expression of c-met in lymph node metastasis positive group was higher than that in lymph node metastasis negative group(P<0.05);there was no significant difference between squemous cell carcinoma group and adenocarcinoma group（P>0.05）;there was also no significant difference between clinical stage Ⅰ group and clinical stage Ⅱ group.In chronic cervicitis,the PCNA index was 24.79%±5.10%,but in cervical cancer the PCNA index was 39.90%±15.46%,the difference of PCNA index was significant between chronic cervicitis and cervical cancer (P<0.05).The lower of tissure differentiation degree and higher of clinical stage,the higher of PCNA expression(P<0.05).There was no significant difference of the expression of PCNA between squemous cell carcinoma group and adenocarcinoma group(P>0.05).The PCNA index in c-met expression positive tumor was significantly higher than that in the negative ones(P<0.01). Conclusion The expression of c-met has strong relationship with PCNA,it could serve as a prognostic factor of cervical cancer.

To investigate the expression of monocyte chemoattractant protein-1(MCP-1) in pulp tissue in patitents with pulpitis,and elucidate its role and mechanism in occurrence and development of pulpitis.Methods Using immunohistochemical staining method,the expression and distribution of MCP-1 in 30 cases of normal pulps,30 acute and 30 chronic pulpitis tissues were detected.Results Different extent of positive MCP-1 expression was detected in acute and chronic pulpitis tissues,but no expression was detected in normal pulp tissue.The yellow particles in cytoplasm were the positive expression,which located in leukocytes,monocytes and macrophages in acute pulpitis tissue.In chronic pulpitis tissue the positive cells were vascular endothelial cells,macrophages,leukocytes,and so on.The extent of positive MCP-1 in acute pulpitis tissue was 119.73±17.12 and the extent of positive MCP-1 in chromic pulpitis tissue was 121.26±2.04,the difference was not significant(P>0.05).Conclusion MCP-1 expresses in pulp tissue in patients with pulpitis.MCP-1 could conduce the pulpitis tissue to heal in the occurrence and development of pulpitis.

To investigate the effect and clinical significance of HBV replication on the serum levels of IL-12 and IL-18 in patients with hepatitis B-related cirrhosis.Methods The levels of HBV-DNA and IL-12 and IL-18 in peripheral blood from 136 patients with different clinic types chronic hepatic diseases（102 patients with chronic hepatitis B,34 patients with hepatitis B-related cirrhosis ），95 HBV carriers and 20 healthy people (as control group) were respectively detected with ELISA and real time PCR. Results The levels of IL-12 and IL-18 in chronic hepatitis B and hepatitis B-related cirrhosis groups were significantly increased compared with control group(P<0.05). The levels of IL-12 and IL-18 in high replication groups of serum HBV-DNA(HBV≥10⁵•mL^-1) were markedly higher than those in negative HBV-DNA group(HBV-DNA＜10³•mL^-1,P<0.05)；there was no significant difference of the serum IL-12 and IL-18 levels between chronic hepatitis B group and hepatitis B-related cirrhosis group，and among different HBV carriers（P>0.05）; and there was positive relationship between prevalence of HBV replication and IL-12,IL-18 levels　in different chronic hepatic disease groups（r=0.74,P<0.05;r=0.80,P<0.05）. Conclusion HBV replication can affect the levels of IL-12 and IL-18 in peripheral blood in patients with hepatitis B，patients with hepatitis and B-related cirrhosis, the higher the levels of the HBV replication,the higher the levels of IL-12 andI L-18 in different chronic hepatic diseases.

To study the expressions of RhoC and MMP-2 proteins in epithelial ovarian cancer and their clinical significances in order to provid the basis for assessing the biological behaviour of epithelial ovarian cancer.Methods Immunohistochemistry was used to detect the expressions of RhoC and MMP-2 proteins in 10 cases of normal ovarian tissues,12 cases of epithelial benign ovarian tumor and 91 cases of epithelial ovarian cancer,and the relationship with the clinical features was analyzed.Results The RhoC and MMP-2 proteins positive expression rates in ovarian benign tumor and ovarian normal tissues were 0 respectively; the positive expression rates of RhoC and MMP-2 proteins in epithelial ovarian cancer were 37.3% and 68.1%.The expressions of RhoC and MMP-2 proteins in ovarian cancer were significantly higher than those in ovarian benign tumor and ovarian normal tissues.The positive expression rates of RhoC and MMP-2 in moderately and poorly differentiated cancer were significantly higher than those in well differentiated cancer(P<0.005).The positive expression rates of RhoC and MMP-2 in Ⅲ-Ⅳclinical stage cancer were significantly higher than those inⅠ-Ⅱstage cancer(P<0.005 ).The expressions of RhoC and MMP-2 proteins in ovarian cancer with metastasis were significantly higher than those in ovarian cancer without metastasis(P<0.005,P<0.05).The expression of RhoC had positive correlation with the expression of MMP-2 in ovarian cancer(r=0.492,P<0.05).Conclusion The expressions of RhoC and MMP-2 proteins show positive correlation with the cell differentiation,clinical stage and metastasis of ovarian cancer.The clinical detection of RhoC and MMP-2 protein expressions in ovarian cancer can be used as a indicator to judge the biological behavior and prognosis of ovarian cancer.

To explore the relationship between expression of muc-1 gene in non-small cell lung cancer (NSCLC) tissues and lymph node metastasis and its clinical significance.Methods The expression of muc-1 gene in 36 specimens of NSCLC tissues and 10 normal lung tissues were determined by RT-PCR.Results muc-1 gene was positively expressed in 19 of 36 specimens of NSCLC tissues,the positive expression rate was 52.7%.The expression of muc-1 mRNA in NSCLC tissues was related to metastasis of lymph node(χ² =6.733,P<0.05),not related to age,sex,histological types,pathological grades,TNM stages,tumour size of the patients（P>0.05）.The positive expression rate of muc-1 gene was higher in NSCLC tissues with lymph node metastasis（N₀　10%,N₁　58.82%,N₂　88.9%）. Conclusion The lymph node metastasis rate in NSCLC patients with muc-1 gene positive expression is higher,the detection of muc-1 gene expression can guide the prognosis and therapy of NSCLC patients after operation.

To compare the immunoreactions between hepatitis B vaccine made by recombinant yeast-derived hepatitis B and CHO hepatitis B,and study serological reaction in different vaccines and different dose in order to provide the best hepatitis B vaccine dose in adults. Methods 238 college students were randomly divided into 4 groups to receive yeast 10 μg,CHO 10 μg,CHO 20 μg and yeast 20 μg at 0,1,6 months,the immune effects including the positive inversion rate of serum anti-HBs and the GMT of anti-HBs and adverse events were observed. Results The positive inversion rate of serum anti-HBs was 98.3% in yeast 10 μg group,96.7% in CHO 10 μg group,95.0% in CHO 20 μg group and 96.5% in yeast 20 μg group and there was no significant difference between each group (χ²=1.034,P>0.05).There were significant differences of the GMT of anti-HBs between four groups（Kruskal-Wallis χ² =18.301,P<0.05）.The GMT of anti-HBs in yeast 10 μg group was higher than that in CHO 10 μg group (P<0.05).There was no significant difference of the GMT of anti-HBs between yeast 20 μg and CHO 20 μg groups(P>0.05).The GMT of anti-HBs in CHO 20 μg group was higher than that in CHO 10 μg group (P<0.05),there was no significant difference between 10 μg yeast group and 20 μg yeast group（P>0.05）,there was no significant difference between the positive inversion rate of serum anti-HBs and sex(Mann-Whitney U=6　512.000，P>0.05).Conclusion Good immunity could be obtained when 10 μg recombinant hepatitis B vaccine is vaccinated in adults.

To approach the correlation between the bone density measured by quantitative CT and the blood sugar level of the aged patients with non-insulin-dependent diabetes mellitus，and observe the effects of the blood sugar level on the bone density.Methods The lumbar bone densities and the blood sugar levels of 160 aged patients with non-insulin-dependent diabetes mellitus (hyperglycemia group 80 cases,euglycemia group 80 cases ) and the healthy aged people (80 cases) were detected by quantitative CT and serum biochemical detection; the correlation between the blood sugar level and the bone density and the osteoporosis occurrence status of aged people in various groups were analyzed.Results The bone density in the non-insulin-dependent diabetes and hyperglycemia group was lower than those in normal(control) group and non-insulin-dependent diabetes and euglycemia group (P<0.05); the morbility of osteoporosis in the non-insulin-dependent diabetes and hyperglycemia group was higher than those in normal(control) group and non-insulin-dependent diabetes and euglycemia group (P<0.05);negative correlation was found between the bone density and the blood sugar level (aged male group：r=－0.7382，P=0.0013；aged female group：r=－0.8343，P=0.0007).Conclusion The blood sugar level affects the bone density of the aged patients with non-insulin-dependent diabetes mellitus;the higher the blood sugar level,the lower the bone density.The non-insulin-dependent diabetes aged patients with hyperglycemia have the liability of osteoporosis.

To investigate and evaluate the detection and the variety of histology type constituent of the thyroid malignant tumor before and after universal salt iodization.Methods 1011 clinical pathological data of thyroid malignant tumor confirmed pathologically from 1961 to 2000 was retrospectively analyzed.The detection rate of thyroid malignant tumor,the constituent ratios of each histology type and the changes of age and sex distribution in main types of thyroid malignant tumor were determined.Results The total detection rate of thyroid malignant tumor after universal salt iodization (USI) (0.69%) were obviously increased compared with before universal salt iodization(0.46%,P<0.01).The constituent ratios of thyroid malignant tumor showed differences before and after universal salt iodization (P<0.01).The proportion of papillary carcinoma (70.17%) increased obviously after USI compared with before USI (55.84%).The proportion of follicular carcinoma (11.08%) decreased accordingly after USI compared with before USI (24.58%).The average age of thyroid malignant tumor sufferers (43.06±13.09 years old) after USI increased(P<0.05) compared with before USI (40.95±14.71 years old).The peak ages of papillary carcinoma(PC),follicular carcinoma (FC) and undifferentiated carcinoma(UC) were increased after USI(＞40 years old) than before USI(≤40 years old).The incidence rates of thyroid　malignant tumor in female patients were higher than male patients before and after USI.Conclusion The proportion and average age of thyroid malignant tumor increases after USI.The histological types of thyroid carcinoma have changes after USI: the proportion of PC increases obviously,the proportion of FC decreases accordingly.The average age of thyroid malignant tumor sufferers tends to increase and the peak ages of PC,FC and UC raise after USI.