Abstract:Objective
To construct the pshuttle-Egr-1-hSmac plasmid and transfect human breast cancer MDA-MB-435 cells, and to observe its radiotherapy enhancing effect on tumor cells.Methods The empty vector pshuttle and pshuttle-Egr-1-hSmac plasmid were transfected into MDA-MB-435 cells by liposomal.At different time（4,8,12,24 and 48 h） after irradiation with 2.0 Gy X-ray and 24 h after irradiation with 0.5―5.0 Gy,the total RNA and protein were collected and extracted from these cells to analyze the Smac mRNA and protein expression levels with RT-PCR and Western blotting methods.The cells were divided into control,pshuttle,pshuttle-Egr-1-hSmac,2.0 Gy irradiation group,pshuttle + 2.0 Gy irra diation and pshuttle-Egr-1-hSmac+2.0 Gy irradiation groups.MTT method was used to evalua te cell proliferation,and the cell survival ability  was measured with clone formation assay;Annexin Ⅴ/PI double staining and PI single staining were used to examine the  apoptosis and cell cycle of MDA-MB-435 cells.Results There was no Smac mRNA expression in MDA-MB-435 cells in control and pshuttle groups,but the Smac mRNA expression levels in  MDA-MB-435 cells in  pshuttle-Egr-1-hSmac plasmid group were gradually increased with the time prolongation,and reached the maximum at 24 and 48 h;the Smac mRNA expression levels in MDA-MB-435 cells  were increased gradually 24 h after irradiation of 0.5―5.0 Gy X-ray with the increasing of irradiation doses,and reached the maximum after 2.0 and 5.0 Gy irradiation.The Smac protein expr ession levels in  pshuttle-Egr-1-hSmac plasmid group were increased gradually with the time prolongation,and reached the maximum at 24 h.The Smac protein expression lervels were increased 24 h afer irradiation of 0,0.5,1.0,2.0 and 5.0 Gy X-ray,especially in 5.0 Gy group.The MTT results showed that the  A490 values  in 2.0 Gy,pshuttle+2.0 Gy and    pshuttle-Egr-1-hSmac groups 24,48,and 72 h after  irradiation were lower than th ose in control group(P<0.01);the  A490 values  of MDA-MB-435 cells in  pshuttle-Egr-1-hSmac group after 1.0-5.0 Gy X-ray irradiation were lower than those in 0 Gy group (P<0.05 or P<0.01);the survival fraction(SF) in pshuttle-Egr-1-hSmac group was lower than those in control group(P<0.01). The percentages of the cells at G0/G1 and S phase in pshuttle-Egr-1-hSmac group were lower than those in 2.0 Gy group(P<0.01)，the percentage of the cells at G2/M phase was higher than that in 2.0 Gy group (P<0.01);the apoptotic rate of the cells  in pshuttle-Egr-1-hSmac group was higher than that in 2.0 Gy group (P<0.01).Conclusion X-ray irradiation can significantly increase the Smac mRNA and protein expression levels in MDA-MB-435 cells transfected with pshuttle-Egr-1-hSmac plasmid,inhibit the  cell survival rate,and  induce G2/M arrest and apoptotic increasing;Smac gene combined with radiotherapy could  significantly increase the  radiosensitivity of breast cancercells.

Abstract:Objective
To explore the role of metabotropic glutamate receptors (mGluRs) group Ⅲ and its subtypes mGluR7 and mGluR8 in nucleus tractus solitarius(NTS) in cardiac-somatic motor reflex (CMR), and to clarify the modulation role of mGluR Ⅲ and its subtypes in NTS in cardiac nociceptoion.Methods 40 SD rats were randomly divided into L-AP4 group,microinjection of mGluRs Ⅲ agonist L-AP4 0.1,1.0,10.0 or 20.0 nmol in NTS；AMN082 group, microinjection of mGluR7 agonist AMN082 1,2 or 4 nmol；DCPG group,  microinjection of  mGluR8 agonist DCPG 4，6 or 8 nmol;MSOP group,microinjection of   mGluR Ⅲ  antagonist MSOP 20 or 100 nmol，20 nmol MSOP+410 nmol L-AP,20 nmol MSOP+2 nmol AMN082,20 nmol MSOP+6 nmol DCPG.The changes of CMR of the rats in various groups were observed.Results Compared with control,the CMR in L-AP4 and AMN082 groups was decreased (P<0.05);the CMR in DCPG group was increased (P<0.05)； the CMR in   MSOP group after injection of 20 nmol MSOP had no change (P>0.05);the CMR in   MSOP group after injection of 100 nmol MSOP was increased (P<0.05);the CMR in MSOP group after injection of 20 nmol MSOP followed L-AP4 or AMNO82 had no change (P>0.05).Conclusion The  group Ⅲ  mGluRs in the NTS play an inhibitory role in cardiac nociception,and mGluR7 has anti-nociceptive effects while mGluR8 has pro-nociceptive effects.

Abstract:Objective
To observe the influence of curcumin in the levels of  nitric oxide (NO) and S100βin the brain tissue of the  rats after cerebral ischemia reperfusion,and to explore the neuroprotective mechanism of curcumin.Methods Adult male SD rats were randomly divided into sham operation group,model group and curcumin group,and 18 rats in every group.Curcumin was intraperitoneally injected daily for 3 consecutive days before inducing focal cerebral ischemia in the SD rats by middle cerebral artery occlusion(MCAO) for 2 h.TTC staining was used to observe the infarction volume.Nitrate reductase assay was used to detect the level of NO in brain tissue of the rats.The level of S100β in brain was detected by ELISA method.Results Compared with model group,the  brain infarction volumes of the rats 24 and 72 h after cerebral ischemia reperfusion in curcumin group were significantly decreased (P<0.05).Compared with sham operation group,the NO and S100β levels in the brain tissue 24 and 72 h after cerebral ischemia reperfusion of the rats in model group were significantly increased(P<0.05);compared with model group,the levels of NO in the brain tissue 24 and 72 h after cerebral ischemia reperfusion in curcumin group were remarkably decreased (P<0.05）;compared with  modee group,the level of S100β in the brain tissue 72 h after cerebral iscemia reperfusion   in curcumin group was remarkably decreased  (P<0.05).Conclusion Curcumin can significantly reduce the degree of ischemia reperfusion injury in the rats and reduce the levels of NO and S100β in brain tissue,which suggests that the decrease of NO and S100β levels in brain tissue may be associated with the neuroprotective effect of curcumin.

Abstract:Objective
To investigate the influence of  truncated apoptosis inducing factor (AIFΔ1-480) on the proliferation and invasion of MCF-7 cells, and to clarify the possibility of promoting cancer gene-radiotherapy. Methods The human breast cancer MCF-7 cells were transfected with  AIFΔ1-480 recombinant expression vector pcDNA3.1-Egr-1- AIFΔ1-480 (pE-AIFΔ1-480) mediated by Egr-1;24 h after 2 Gy X-ray irradiation,MTT assay and Transwell invasion assay were performed to measure the changes of cell proliferation and invasion.The MCF-7 cells were diveded into normal control,pcDNA3.1,pE-AIFΔ1-480,2 Gy irradiation and pE-AIFΔ1-480 + 2 Gy irradiation groups.Results After transfection and 2 Gy X-ray  irradiation,the cells proliferated very fast in normal control,pcDNA3.1 and pE-AIFΔ1-480 groups,and the proliferation regularity was similar.Compared with normal control group,the cell proliferation abilities were  significantly decreased in 2 Gy irradiation and pE-AIFΔ1-480 + 2 Gy irradiation groups (P<0.05),and it was more obvious in pE-AIFΔ1-480 + 2 Gy irradiation group,and it was significant lower than that in 2 Gy irradiation group (P<0.05).The number of the cells permeating membrane was basically same in normal control,pcDNA3.1 and pE-AIFΔ1-480 groups; compared with normal control group,they were significantly decreased in 2 Gy irradiation and pE-AIFΔ1-480+ 2 Gy irradiation groups（P<0.05 or P<0.01）；and it was more significant in pE-AIFΔ1-480 + 2 Gy irradiation group than that in 2 Gy irradiation group (P<0.01).Conclusion AIFΔ1-480 and ionizing radiation could inhibit the proliferation and invasion of human breast cancer MCF-7 cells,both of them have a synergistic effect,and Egr-1 promoter can enhance the suppression effect under radiation conditions.

Abstract:Objective
To establish a nerve stem cell model possessing the potentiality of   differentiating into γ-aminobutyric acid neuron-like cells(GABAergic-like cells)， and provide eligible investigative vector for study on  GABAergic neurons degenerative diseases. Methods Dibutyryl cyclic adenosine monophosphate (dbcAMP) was applied to induce human neuroblastoma SH-SY5Y cells,and the SH-SY5Y cells  were  divided into control group(0 mmol?L-1 dbcAMP),0.3 mmol?L-1 dbcAMP group,0.6 mmol?L-1 dbcAMP group,1.0 mmol?L-1 dbcAMP group and 2.0 mmol?L-1 dbcAMP group;the morphological  changes of SH-SY5Y cells were observed.The Image-Pro Plus 5.0 software was used to measure the length of neurites of the neuron-like cells,and the  percentage of the SH-SY5Y cells with neurites longer than 30  μm  was calculated.The immunofluorescence cytochemistry technique was utilized to test GAD65 positive cells,and the positive rate of immune response was calculated.Results The results of light microscope observation showed that the cells in control group were polygonal,circular or shuttle type with smooth membrane and clear boundaries.With the increasing of the  concentrations  of dbcAMP and the prolongation of time,the morphology of SH-SY5Y cells’ bodies became smaller with longer processes in dbcAMP groups.The cells interwined each other and showed mature neuron phenotype in 1.0 mmol?L-1 dbcAMP group.Compared with control group(31.4%±4.2%)，the  percentages of the cell with  neurites longer than 30  μm in 0.3，0.6，1.0,and 2.0 dbcAMP groups  (40.1%±5.7%,47.5%±6.2%,73.1%±3.2%,and 74.3%±6.1%) 72 h after induction were significantly increased(P<0.05 or P<0.01).Compared with control group(10.2%±2.1%)，the GAD65 positive expression rates in 0.3,0.6,1.0, and  2.0 mmol?L-1, dbcAMP groups(22.1%±2.4%,46.9%±3.2%,70.7%±3.4%,and 72.3%±3.7%)72 h after induction  were significantly increased(P<0.05 or P<0.01).Conclusion The SH-SY5Y cells have the potentiality of differentiating into  GABAergic-like cells,and 1.0 mmol?L-1  is the optimal concentration of dbcAMP.

Abstract:Objective  To construct recombinant  lentiviral vector pSin-EF2-Puro-DAB2IP transfect prostate carcinoma PC3 cells,and to observe its transfection rate and expression level in PC3 cells.Methods The cDNA sequences specifically targeting the DAB2IP gene were designed and cloned into lentiviral vector PC3-pSin using DNA recombinant technique.Using PC3-pSin-EF2 as control,the 293T cells were transfected by Lipofectamine reagent for lentiviral particles packaged，and viral titer was determined.The DAB2IP mRNA and protein expression levels were examined by RT-PCR and Western blotting methods.Results The PCR and sequencing analysis results confirmed that the DAB2IP gene sequence was consistent with the sequence in GeneBank.The number of DAB2IP gene copy in PC3-pSin-EF2-DAB2IP cells and its control PC3-pSin-EF2 cells were 0.001±0.000 and 0.158±0.013,respectively.Compared with control cells,the overexpression of mRNA in PC3-pSin-EF2-DAB2IP cells upregulated (179.370±15.891) times,the difference was statistically significant (P<0.001). Compared with internal reference and control cells,the expression levels of DAB2IP protein in PC3-pSin-EF2-DAB2IP cells upregulated (2.431±0.892) times and (2.415±0.961) times respectively,the differences were statistically significant (P<0.001).Conclusion The lentiviral vector of the DAB2IP gene pSin-EF2-Puro-DAB2IP is successfully constructed，and its targeted gene is stably expressed in the targeted cells,which provides a basis for the further functional study of DAB2IP.

Abstract:Objective
 To establish mouse poisoning model by inhaling benzene,and to investigate the induction effect of benzene on the apoptosis of mouse bone marrow cells and its mechanism,and to provide an experimental basis for study on bone marrow toxicity mechanism.Methods 24 male mice were randomly divided into four groups (n=6).The mice in one group were exposed to ambient air (control group) and the mice in the other three groups were exposed to different doses (400,800,1 600 mg?m-3) of benzene (low,middle and high doses of benzene groups) for 15 d in the respective inhalation chambers.At the end of the  experiment,the mice were killed.The bone marrow of the mice was obtained.The pathological changes of the bone marrow cells of the mice in various groups were observed under light microscope with HE staining.The apoptotic rates and mitochondrial membrane potential (MMP) of the mice in various groups were detected by flow cytometry, and the expressions of mitochondrial-deperdent apoptosis related gene proteins were determined with immunohistochemistry method.Results The number of distal and central cells in different doses of benzene groups were significantly reduced,and accompanied by blood sinus expansion in high dose of benzene group.The apoptotic rates of the cells in middle and high doses of benzene groups were obviously higher than that in ｃontrol group (Ρ<0.01),and there were also significant differences between high dose group and low,middle doses of benzene groups (Ρ<0.05).The MMP was significantly decreased with the increasing of benzene doses,and there were significant differences between middle,high doses of benzene groups and control group (Ρ<0.05).The number  of Bax，CytC positive cells in different doses of benzene groups and the number  of Caspase-9,Caspase-3 positive cells in middle and high doses of benzene groups were significantly increased compared with control group(Ρ<0.05);the number of Bcl-2 positive cells in different doses of benzene groups was decreased(Ρ<0.05),and number of Bcl-2 positive cells in middle and high doses of benzene groups was decreased compared with low dose of benzene group(P<0.05).Conclusion Benzene with certain dose can induce the apoptosis of mouse bone marrow cells,and promote the expressions of mitochondrial apoptosis related gene proteins.Benzene-induced apoptosis through mitochondrial-dependent apoptosis pathway may be an important mechanism of bone marrow toxicity induced by benzene.

Abstract:Objective
To explore the effects of tanshinone ⅡA on the injury of dopaminergic neurons of Parkinson’s disease (PD) mouse model induced by 1-methy-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP),and to clarify the possible mechanism of its protective effects on dopaminergic neurons.Methods 60 C57BL/6N mice were randomly divided into control group,PD  model group and tanshinone ⅡA group(n=20).The mice in PD model group and tanshinone ⅡA group were treated with MPTP to establish PD models.The bebavior of the mice  in various groups was observed.The number of tyrosine hydroxylase (TH),cd11b,p47-phox,and inducibleni tricoxidesynthase (iNOS)-positive cells,and the protein expression levels of TH,cd11b,p47-phox and iNOS in the substantia nigra (SN) of the midbrain of the mice in various groups  were detected with immunohistochemistry,Western blotting and double-labeling immunofluorescence assay. Results Compared with control group,the mice  in PD model group exhibited typical symptoms of PD,and the number of TH-positive cells and the protein expression level of TH in the substantia nigra of the mice in PD model group were reduced by about 45% and 50%;the number of cd11b,p47-phox,iNOS-positive cells and the protein expression levels of cd11b,p47-phox,iNOS were markedly increased （P<0.01）.Compared with PD model group,the symptoms of PD of the mice in tanshinone ⅡA group were alleviated, the number of TH-positive cells and the expression level of TH protein in the SN were increased(P<0.01),and the number of cd11b,p47-phox and iNOS-positive cells and the TH protein expression levels were obviously decreased（P<0.01）.Conclusion Tanshinone ⅡA could mitigate the loss of dopaminergic neurons in the PD mouse model induced by MPTP.The mechanism of neuprotective effect may be related to the inhibition of microglial activation,NADPH oxidase and iNOS expressions.

Abstract:Objective
To discuss the influence of eldepryl on the expressions of glial fibrillary acidic protein (GFAP) and cd11b in substantia nigra and striatum in the rats with Parkinson’s disease (PD),and to clarify the regulatory role of eldepryl in the gliacytes.Methods 72  SD rats were randomly divided into control group,PD model group and eldepryl group,and each group was divided randomly into 4 d and 8 d subgroups (n=12) after the success of model preparation.The PD rat models were established by injecting rotenone in subcutaneous.The number of GFAP and cd11b positive cells and the expressions of GFAP and cd11b were detected by immunohistochemistry and Western blotting method．Results The GFAP and cd11b positive cells were all in a resting state in control group,the GFAP-positive cell body was slender and irregular and had elongated protrusions;the cd11b-positive cell body was small and   branch-like,and it had more slender protrusions.The GFAP and cd11b positive cells were all in a active state in model group,the GFAP-positive cell body was hypertrophy,the projections increased thickening;the cd11b-positive cell body was more bigger,the projections were shorter and thicker,and the number was increased.Compared with model group,the GFAP-positive cell body and protrusions were more slender,the CD11b-positive cell body was more smaller,the projections were more slender,and the number was decreased in eldepryl group.There were a small amount of expression of GFAP and cd11b positive cells in substantia nigra and striatum in the rats in control group,and there was no significant difference between 8 d group and 4 d group(P>0.05). The number of GFAP and cd11b              positive cells and the protein expression levels were significantly increased in model group compared with  control group(P<0.01);there was more expression in 8 d group compared with 4 d group, but there was no significant difference(P>0.05).The number of GFAP and cd11b positive cells and the protein expression levels  in eldepryl group were significantly reduced compared with model group(P<0.01);there were less expression in 8 d group compared with 4 d group,and there was significant difference(P<0.05).Conclusion There are activation and proliferation of the gliacytes in substantia nigra and striatum in the rats with PD,and eldepryl can inhibit the activation and proliferation of gliacytes.

Abstract:Objective
To explore the influence of intraventricular injection of 5,7-drhydroxytryptamine(5,7-DHT) in  5-HT1A receptor sensitivity of medial prefrontal cortex  pyramidal neurons in the  rats,and to clarity the effect of 5-HT1A receptor on the eletronic response of pyramidal neurons.Methods 36 male SD rats were randomly divided into sham operation group (n=21) and 5,7-DHT lesion group (n=15).5,7-DHT was injected  intraventricularly in the rats  in 5,7-DHT lesion group,and the same dose saline was injected in the rats in sham operation group.The rats in two groups were intravenously injected with different doses(0.5-128.0  μg•kg^-1) of 8-CH-DPAT.The firing rate  of mPFC pyramidal neurons was recorded with　extracellular　electrophysioological examination.The rats in two groups were intravenously injected with WAY100635,the sensitivites of the rats to 8-OH-DPAT and WAY100635 in 5,7-DHT lesion group were observed,and compared with  sham operation group.Results The different doses (0.5-128.0  μg•L-1) of 8-OH-DDAT had an excitatory-inhibitory effect on the firing rate of mPFC pyamidal neurons in sham operation group;the neurons were excited when the doses of 8-OH-DPAT were 0.5-38.0  μg•kg^-1,and the firing rates were increased(P<0.05);the neurons were inhibited when the dose of 8-OH-DPAT was 128.0  μg•kg^-1,and the firing rate was decreased.The different doses(0.5-218.0  μg•L^-1) of 8-OH-DPAT inhibited the elecctronic response of pyramidal neurons of the rats in 5,7-DHT lesion group in a dose-dependent manner (df=5,F=3.44，P=0.003),and the firing rates were reduced.WAY-100635 (50 μg•kg^-1) reversed completely the inhibition of 8-OH-DPAT.Conclusion The sensitivity of 5-HT1A receptor of rat mPFC pyramidal neurons  can be  decreased by intraventricular injection of 5,7-DHT.

Abstract:Objective
To investegate the effect of ginsenoside Rg1 on the apoptosis related protein  FLICE- inhibitory protein(FLIP),Fas-associated death domain protein (FADD) and Caspase-3 in the subatania nigra(SN) of 1-methyl-4-phenyl-1,2,3,6-tetrahyd- ropyridine (MPTP)-induced mouse models of Parkinson’s disease(PD),and to investigate the role of  FADD  and FLIP in the pathogenesis of PD and the protective effect of ginsenosides Rg1 on dopaminergic neurons.Methods 45 C57BL/6N mice were randomly divided into control group,  model group and ginsenoside Rg1 group (n=15).The mice in model group were injected with MPTP by intraperitoneal,the mice in Rg1 group were injected with ginsenoside Rg1 before injecting MPTP,and the mice in control group were injected with normal saline by intraperitoneal.The behavioral changes of the mice in various groups were observed,and immunohistochemistry and Western blotting methods were used to observe the expressions of tyrosine hydroxylase(TH),FADD,FLIP and Caspase-3 in substantia nigra of the mice.Results Compared with control group,the mice in model group presented with typical symptoms of PD,the TH-positive neurons in the subatania nigra was significantly reduced (P<0.01),the number of  FADD,FLIP and Caspase-3 positive cells  was significantly increased(P<0.01),and the cytoplasm was deeply stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly increased (P<0.01).Compared with model group,the PD symptoms of the mice in ginsenoside Rg1 group reduced,the number of TH-positive neurons was significantly increased,the number of positive cells of FLIP,FADD and Caspase-3 were significantly reduced(P<0.01),and the cytoplasm was lightly stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly reduced (P<0.01).Nonlinear correlation analysis found that there was a positive  relationship between the number of FADD and  Caspase-3  positive cells  (r=0.791,P<0.05).Conclusion Ginsenoside Rg1 may play a neural protective effect dopaminergic on neurons  by modulating the   FADD and FLIP expressions in SN of PD model mice.

Abstract:Objective
To observe the influence of sirtinol，a silent information regulator 1(SIRT1) inhibitor,in the cell proliferation,cell cycle progression and the expression levels of positive regulator proteins of the cell cycle including Cyclin D1，CDK4  and pRb  in prostate cancer DU145 cells,and to explore the possible mechanism of SIRT1 in  occurrence of prostagte cancer.Methods The DU145 cells at logarithmic growth phase were cultured in vitro and divided into control group（DMSO）and different doses (10,25,50  μmol?L-1) of sirtinol  groups.The inhibitory rate of  growth of DU145 cells was detected with MTT method,the SIRT1 mRNA and protein expression levels were determined by RT-PCR and Western blotting method,and the cell cycle was measured by flow cytometry.The Cyclin D1，CDK4 and pRb protein expression levels were examined by Western blotting method.Results Compared with control group，the inhibitory rates of growth of the DU145 cells in different doses of sirtinol groups were increased markedly in a dose-dependent manner（P<0.01）,and the flow cytometry analysis result showed the DU145 cells at G1 phase were increased（P<0.01）.Compared with control group，the expression levels of SIRT1 mRNA and protein in DU145 cells in different doses of sirtinol groups were decreased significantly（P<0.01）;the expression levels of  Cyclin D1 and pRb proteins were decreased（P<0.01）,whereas the expression levels of CDK4 had no change（P>0.05）.Conclusion SIRT1 inhibition by sirtinol can inhibit the cell growth of prostate cancer DU145 cells in a dose-dependent manner and arrest the cell cycle progression，and its mechanism may be related to decreasing the CyclinD1 and pRb protein expressions.

Abstract:Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt) inhibitor LY294002, and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group.The inhibitory rate of  growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis.The expression levels of Ser473 phosphorylated form of Akt(p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method.Results The inhibitory rate of  growth of the A549 cells in LY294002+rsTRAI group (74.6%±2.63%)was higher than that in rsTRAIL group (5.61%±0.32%) (P<0.05).Compared with rsTRAIL group,the percentage of the cells at G0/G1 phase  in LY294002+rsTRAIL group was increased(P<0.05) and the percentage  of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%) was higher than that in rsTRAIL group (3.21％±0.96％) (P<0.05). The Western blotting results showed that the expression levels of p-Akt,c-FLIPL  and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05) and the  ratio of Bax/Bcl-2 was increased (P<0.05) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.

Abstract:Objective To investigate the influence of  Schizandrain B(Sch B) preconditioning in myocardial cells of neonatal rats with hypoxia / reoxygenation (H/R) injury,and to explore its possible mechanism.Methods The cultured myocardial cells were divided into control group,model group  and 10,50,250 mg/LSchB preconditioning  groups.All the cells in various groups except control group were cultured in 2 mmol/L Na2S2O4  for 2 h，then cultured in normal medium for 18 h to induce myocardial H/R injury.The morphological changes of myocardial cells in various groups were observed under inverted microscope.The survival rates of the myocardial cells in each group were  examined  by MTT.The activities of lactic dehydrogenase(LDH),creatine(CK),superoxide ismutase(SOD),and the (MDA) levels in the cells in various groupswere  examined  by detection kits.Results Compared with control group,the cells in model group were retracted,arrest or float,the survival rate was decreased significantly (P<0.01),the activities of LDH,CK and MDA level were increased (P<0.01),and the SOD activity was decreased (P<0.01);compared with model group,the cells in Sch B preconditioning groups  remained beating,retraction was light,the cell survival rates were significantly increased (P<0.01),the activities of LDH,CK and MDA levels were decreased (P<0.01),and the SOD activities were increased (P<0.05 or P<0.01).Conclusion Sch B has protective effect on  myocardial cells with H/R injury in the neonatal rats,which may be associated with anti-oxidative damage.

Abstract:Objective To observe the influence of rizatriptan on the behavior and pain-related cytokines in peripheral blood of the  migraine model rats,and to investigate the theraputic effect of rizatriptan on migraine.Methods A total of 24 rats were randomly divided into: control group,migraine  group,rizatriptan control group and rizatriptan treatment group.The rats in rizatriptan control and rizatriptan treatment groups were intragastrically perfused with rizatriptan,1 mg/kg per day (according to the adult daily dose),and the rats in  control and migraine  groups were perfused with normal saline, 1 mL per day.After 7 d,nitroglycerin was subcutaneously injected into the buttocks of the rats in rizatriptan treatment and migraine  groups to induce migraine.Normal saline  was injected into the rats in  control and rizatriptan control groups.At 60-90 min following nitroglycerin injection,the total number of behavioral symptoms was measured.The serum calcitonin gene-related peptide(CGRP),5-hydroxytryptamine(5-HT),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α) levels were determined using ELISA.Results Compared with migraine  group,the behavioral score of the rats in rizatriptan treatment group was significantly decreased (P<0.05). The serum CGRP level of the rats in migraine  group was significantly higher than that in  control group (P<0.05).Compared with  control group,the serum 5-HT level of the rats  in  and migraine  group was   decreased,but there was no significant differece(P>0.05);the serum 5-ＨＴ level in rizatriptan control group was significantly increased(P<0.05).The serum 5-HT level of the rats in rizatriptan treatment group was significantly increased compared with migraine  group (P<0.05).The serum IL-1β and TNF-α levels of the rats in varions groups had no significant differences (P>0.05).Conclusion Rizatriptan can relieve the  behavior symptoms in nitroglycerin-induced  migraine model rats,increase the  serum 5-HT level,improve the vasomotor disturbance,and relieve the angiectasis.

Abstract:Objective To observe the influence of XueShuanXinMaiNing（XSXMN） in the behavior and structures of cerebral cortex and hippocampus  of the rats with β amyloid protein(Aβ）-induced Alzheimer’s disease(AD),  and to explore its therapeutic effects on the rat  AD.Methods 100 male Wistar rats were selected.According to weight,the rats were randomly divided into sham operation group,model group,positive drug group(donepezil hydrochloride,1.75 mg/kg),XSXMN 1.1 g/kg group and XSXMN 2.2 g/kg group. The  rat AD models  were made by injecting Aβ into hippocampus.After oral administration for 15 d,Morris water maze test,dark avoidance task and pathology test were performed.Results In  Morris water maze test,compared with model group,the latency and swimming distance to platform of the rats in XSXMN 1.1 g/kg group were decreased on the 2nd,4th and 5th day（P<0.05 or P<0.01）;in XSXMN 2.2 g/kg group,the latency to platform of the rats were decreased from the 3nd to 6th day（P<0.05 or P<0.01）,the swimming distances to platform of the rats were decreased from the 3rd to 5th day(P<0.05 or P<0.01).On the 7th day,in  XSXMN groups,the times of passing platform,time of staying on platform,distance of staying on platform,time of staying in effective area,distance of staying in effective area,time of staying on platform/total time,distance of staying on platform/total distance,time of staying on platform/total time were all increased significantly（P<0.05 or P<0.01） within 90 s.In dark avoidance task,compared with model group,the error latency and the error times of the rats in XSXMN groups had no obvious change on the 2nd day.The pathological results showed that there were degeneration nerve cells and necrosis nerve cells in the rat cerebral cortex in XSXMN groups,while in the rat hippocampus there were less number of nerve cells with obscure cell layer and many degeneration and necrosis cells were found;compared with model group,there was no obvious improvement.Conclusion  XSXMN can improve the learning and memory function of the AD rats.

Abstract:Objective To observe the improvement effects of astragaloside Ⅳ (AS-Ⅳ) on the myocardial focal ischemia-reperfusion (I/R) injury and its influence in PI3K/Akt/mTOR pathway,and to clarify the protective effect of AS-Ⅳ on myocardial I/R injury and the possible mechanisms.Methods The left main coronary arteries of 60 Sprague-Dawley rats  were occluded for 30 min followed by a 120-min reperfusion to induce I/R model.The rats with I/R injury were randomly divided into model group (normal saline),AS-Ⅳ group (intravenous injection of 10 mg/kg AS-Ⅳ 5 min before reperfusion),PI3K inhibitor Wortmannin (WOR) group (intravenous injection of 0.6 mg/kg WOR 10 min before reperfusion) and AS-Ⅳ+WOR group (intravenous injection of   10 mg/kg AS-Ⅳ and 0.6 mg/kg WOR  5 and 10 min before reperfusion,respectively).15 age-matched SD rats were chosen as control group.The heart  mass,degrees of infarction and ischemia and cardiac function ,including left ventricular systolic mean pressure (LVSP),end-diastolic pressure (LVEDP),fractional shortening (FS) and ejection fraction (EF), of the rats in all groups were analyzed.Western blotting method was used to measure the phosphorylation levels of Akt and mTOR(p-Akt and p-mTOR).The specific fluorescent probe DHE staining was employed to detect the myocardial reactive oxygen species levels.Results Compared with control group,the degrees of infarction and ischemia,LVEDP,myocardial levels of p-Akt/Akt,p-mTOR/mTOR and reactive oxygen species levels of the rats were increased (P<0.05). and the levels of LVSP,FS and EF were decreased in model group (P<0.05). Compared with the model group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species level were decreased (P<0.05),while the levels of p-Akt/Akt,p-mTOR/mTOR LVSP,FS and EF of all rats in AS-Ⅳ group were increased (P<0.05).Compared with AS-Ⅳ group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species levels of the rats in   WOR group and AS-Ⅳ+WOR group were increased(P<0.05),and the myocardial  p-Akt/Akt,p-mTOR/mTOR and  LVSP,FS,EF were decreased (P<0.05).Conclusion AS-Ⅳ has improvement effect on myocardial I/R injury.AS-Ⅳ can reduce the extent of myocardial infarction and oxidative stress and improve the heart function,and its possible mechanism may be related to activating PI3K/Akt/mTOR signaling pathway.

Abstract:Objective To investigate the protective effects of Xuebijing(XBJ)on  the injury of vascular endothelial cells(VEC) induced by lipopolysaccharide (LPS), and to study the mechanisms of the production of nitric oxide (NO) and the expressions of inducible nitric oxide sytnhase (iNOS) and signal transduction under XBJ intervention condition. Methods   The cultured VEC were divided into control group, LPS(1 mg?L-1)group, LPS(1 mg/L)　+XBJ(25 g/L)group,LPS(1 mg/L)+pyrrolidine dithiocarbamate(PDTC, 20 μmol/L)group; XBJ and PDTC were  administrated 1 h before incubation of with LPS. Western blotting method was used to detect the expressions of iNOS and NF-κB p65 protein. The level of  NO in the supernatant was measured by Griess reagent. Results   Comparaed with control group, the NO level and the expression levels of iNOS protein and NF-κB p65 protein in VEC in LPS group were significantly increased(P<0.01). Compared with LPS group, the NO level and the expression levels of iNOS protein and NF-κB p65 protein in VEC in LPS+XBJ group were significantly decreased (P<0.05 or P<0.01); the NO level and the expression levels of NF-κB p65 protein and iNOS protein in VEC in LPS+PDTC group were significantly decreased(P<0.05 or P<0.01). There were no significant differences of  the NO levels　 and the expression levels of iNOS protein between LPS+XBJ group and LPS +PDTC group (P>0.05), but the expression level of NF-κB p65 protein in LPS+PDTC group was  lower than  that in LPS+XBJ  group(P<0.05). Conclusion  XBJ can inhibit the  production of NO and the expression of iNOS protein  in VEC; its  mechanism may be related to  inhibiting the activation of NF-κB to control inflammation.

Abstract:Objective To investigate the influence of atorvastatin in methylation and expression level of Bcl-2 in human umbilical endothelial cells(HUVECs) treated with homocysteine(Hcy) and to expound potential mechanism of atorvastatin resisting arteriosclerosis.Methods After HUVECs were treated with 0,2,4,8,16,and 32 mmol/L  Hcy for 48　h,MTT was used to measure the inhibitory rates of HUVECs and the half inhibitory concentration (IC50).According to the experimental results,the HUVECs cultured in vitro were divided into control group(0.00 mmol/L Hcy),Hcy group(9.00 mmol/L Hcy)，and atorvastatin group(9.00 mmol/L Hcy＋１×10-3 mmol/L atorvastatin).After treated for 48 h,flow cytometry was used to detect the apoptotic rate of cells,the mRNA expression of Bcl-2 was analyzed by fluorescence quantitative PCR,the protein expression of Bcl-2 was detected by Western blotting method,and the methylation level of Bcl-2 promoter region was determined by nest touch-down PCR combined with methylation specific PCR(MSP).Results Compared with control group,the apoptotic rate of HUVECs in Hcy group was increased(P<0.01),the mRNA and protein expression levels of Bcl-2 were significantly decreased(P<0.01),and the Bcl-2 promoter region methylation level was also decreased(P<0.01).Compared with Hcy group,the apoptotic rate of HUVECs in atorvastatin group was decreased(P<0.01),the mRNA and protein expression levels of Bcl-2 gene were increased(P<0.05),and the Bcl-2 promoter region methylation level was also increased（P<0.05).Conclusion Atorvastatin can prevent the apoptosis of HUVECs induced by Hcy through regulating Bcl-2 methylation.

Abstract:Objective To investigate the effect of tannic acid on glomerular mesangial cells (GMC), and to clarify the mechanism of tannic acid in improving the pathological changes of diabetic nephropathy (DN) from the aspect of oxidative stress and micro-inflammation. Methods   The glomerular mesangial cells were treated with glucose (30 mmol/L) or  advanced glycosylation end-products (AGEs) bovine serum albumin(BSA) (250 mg/L) and then different concentrations of tannic acid (10, 20, 40 and 80 μmol/L) were added into the GMC. The cells cultured by normal glucose or treated with BSA were used as control groups and then the level of malonic dialdehyde(MDA）, glutathione peroxidase (GSH-Px ), superoxide Dismutase(SOD), CAT (Catalase) activities and   8-hydroxy-2′-deoxyguanosine(8-OHdG) levels in the culture supernatant 48 h after culture  were determined by colorimetry and ELISA method. The expressions of  intercellular cell adhesion molecule-1(ICAM-1)  protein,monocyte chemotactic protein 1(MCP-1) and  ICAM-1 mRNA in GMC  were detected by immunohistochemical staining and RT-PCR method. Results  Compared with high glucose and AGEs groups, the MDA levels in  tannic acid groups were reduced significantly（P<0.05）; the activities of GSH-Px,  SOD and CAT were increased significantly（P<0.05 or P<0.01）;the 8-OHdG levels in annic acid groups were significantly reduced （P<0.05）. Compared with  high glucose and AGEs groups, the expressions levels of ICAM-1  protein in 40 and 80 μmol/L tannic acid  groups were decreased(P<0.05). The mRNA expressions levels  of MCP-1 and ICAM-1 were significantly lower  than those in  high glucose group （P<0.01）.  Conclusion  Tannic acid could protect GMC against the damage of oxidative and inflammatory mediators, thereby delaying and improving the glomerular lesions of DN.

Abstract:ObjectiveTo clarify the inhibitory effect of low molecular weight chitosan（LMCTS）on hepatic fibrosis induced by carbon tetrachloride (CCl4) in the rats,and to investigate its effect on the expression of Toll-like receptor-4(TLR4) and to lay the foundation for the development of the clinical candidate drug of liver fibrosis.Methods 72 male Wistar rats were randomly divided into blank control group,CCl4 group(model group),glycyrrhizinate (DG) group,50,100 and 150 mg/kg LMCTS groups (low,middle and high doses of LMCTS groups).In addition to  blank control group,the rats in the remaining groups were given 40% CCl4-vegetable oil (1.75 mL/kg),2 times per week for 8 weeks,by intraperitoneal injection to establish the model of rat hepatic fibrosis.And the rats in blank control group were injected with the same amount of 100% vegetable oil agent.From the ninth week，the rats in DG and LMCTS groups were given DG and LMCTS by intragastric administration,1 time/week for 4 weeks.Then all rats were sacrificed,the activities of serum glutamic acid aminotransferase (ALT),aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were detected with ELISA kit;the pathological changes in liver tissue were observed under light microscope,and the TLR4 expressions were detected by RT-PCR and Western blotting method.Results The serum ALT,AST and ALP activities in middle and high doses of LMCTS groups were lower than those in model group  (P<0.05).The serum ALT activity in middle dose of LMCTS group was lower than that in low dose of LMCTS group (P<0.05),but the activities of AST and ALP had no  statistically significant change(P>0.05).There were statistically significant differences in the serum ALT,AST and ALP activities  between high dose of LMCTS group and middle  dose of LMCTS group (P<0.05).There were obvious hepatocyte steatosis,inflammatory cell infiltration,collagen fiber hyperplasia and hepatic lobule damage in the rats in model group.However,all the  changes in liver tissue of the rats in LMCTS group  were significantly reduced,especially in high dose group.The results of RT-PCR and Western blotting method showed that the expression of TLR4 was declined in LMCTS groups compared with model group (P<0.05,P<0.01),especially in high dose of LMCTS group,and there were statistically significant differences between  different doses of LMCTS groups (P<0.05).Conclusion High dose of LMCTS can decrease the serum transaminase activity of liver fibrosis rats and improve liver function,and this process may be related to declining the expression of TLR4.

Abstract:Objective To explore the antibacterial activity of extract of Ginkgo biloba leaves on  Porphyromonas gingivalis in vitro , and to provid  pharmacological reference    for developing a new type of antibacterial drugs in the treatment of periodontal disease. Methods  This experiment was divided into negative control group, imipenem  control group and different concentrations and  forms of extract of Ginkgo biloba leaves groups. Solvent extraction method was used to extract the extract of Ginkgo biloba leaves,punching method and test tube method were performed to detect the antibacterial activity of extract of Ginkgo biloba leaves in anaerobic environment in vitro and compared with Staphylococcus  aureus and E.coli. By observing the antibacterial ring diameter and determination of the minimum bacteriostasis concentration (MIC),  the antibacterial activities of extract of Ginkgo biloba leaves in vitro were measured.Results  In the experiment of bacteriostatic ring,Porphyromonas gingivalis was treated with  extract of Ginkgo biloba leaves, Ginkgo biloba leaf tablet and Ginkgo biloba soft capsule  concentrate and 1∶4 diluent,the bacteriostatic ring diameters were decreased with the decreasing  of the concentration.The maximum bacteriostatic diameter  of Ginkgo biloba extract  was 16.5 mm, and the maximum bacteriostasis diameters   of Ginkgo biloba leaf tablet and soft capsule  were 15.3  and 14.5 mm, respectively; the bacteriostatic diameter of the exact of Ginkgo biloba leaves  was bigger than  those  of Ginkgo biloba leaf tablet and Ginkgo biloba soft capsule (P < 0.05), but there  was no statistical difference of bacteriostatic diameter between Ginkgo biloba leaf tablet and Ginkgo biloba soft capsule  (P> 0.05); E. coli and Staphylococcus  aureus  groups get the same results.   When the  concentration of extract of Ginkgo biloba leaves  was more  than 1.95 mg/L, there was no growth of Porphyromonas gingivalis  but  E. coli and Staphylococcus aureusa still grew; only  the concentrations of exact of Girkgo biloba leaves were  more than 6.25  and 12.5 mg/L, E. coli and Staphylococcus aureus didn’t  grow; the bacteriostatic effect of extract of Ginkgo biloba on Porphromonas gingivalis was better  than  E.coli   and Staphylococcus aureus.Conclusion  Extract of Ginkgo biloba  leaves has antibacterial effect on Porphyromonas gingivalis.

Objective To observe the synthesis of melanin  and tyrosinase activity of the cultured normal human melanocytes after treated with baicalin and NB-
UVB,and to provide theoretical  evidence for study on the  new therapy method of vitiligo.Methods The normal human menlanocytes cultured in vitro were
 divided into control group,different concentrations of baicalin groups,different doses of NB-UVB group and baicalin combined NB-UVB group.The proliferation rate of melanocytes was measured by MTT assay,the level  of melanin was tested by NaOH pyrolysis,and the tyrosinase activity was measured by dopa oxidization.Results Compared with control group,the proliferation rates in  10-4 mol/L baicalin group and 30 mJ/cm
  NB-UVB group had no significant difference（P>0.05）.Compared with control group,the levels  of melanin and tyrosinase activities in baicalin group,NB-
UVB group and combined group were  increased （P<0.05 or P<0.01）;compared with baicalin group and NB-UVB group,the level  of melanin
and tyrosinase activity  in combined group were increased（P<0.01）. Conclusion Baicalin combined with NB-UVB has better promotion effect on melanin synthesis than used alone.
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Objective To establish and evaluate the method to collect the  rat gingival crevicular fluid （GCF） by using absorbent paper points,and to lay foundati
on for analysis on GCF.Methods 20 healthy male rats were selected and randomly divided into GCF group and saliv
a group.The GCF of  the right upper molar gingival trough  of the rats in GCF group and the saliva of the rats in saliva group were collected by using 15# ab
sorbent paper points.The SDS-PAGE analysis  and abundance detection were applied to  analyze the protein bands of the sa
mples in two groups.Results The SDS-PAGE analysis identified the proteins at 77 000,66 000,55 000,51 000,and 28 000,especially 66 000 in GCF group.While saliva group had lower brightness protein bands at 66 000,60 000,and 48 000.The data of protein abundance of 66 000 between two groups had statistically significant difference (P<0.05). Conclusion The number and types of the protein bands are different between GCF Group and saliva group,so using 15# absorbent paper points can collect  the rat GCF successfully.

Objective To extract the  Ginseng polysaccharide (GPS),polysaccharides of Tricholoma matsutake(PTM) and polysaccharide of Lentinus edodes
 (PLE) from gingeng,tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity in vitro  of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by etha
nol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The  m-hydroxyphenyl method was used to determine the levels of  uronic acid,and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods.Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of  GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.
Results The extraction rates of  GPS,PTM,and PLE were 8.85%,9.40%,and 10.50%; the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;
the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%， and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg/L,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained, identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.

Objective To investigate the association between 5-hydroxytryptamine 2A receptor(HTR2A) gene T102C locus  polymorphism and schizophrenia,and to provide basis for evidence-based medicine for the genetic background of schizophrenia.Methods PubMed,EMbase,CNKI,WanFang and Vip information  databases were used to search full text of all the relevant studies about the association between HTR2A gene T102C locus  polymorphism and schizophrenia,which were published during 2003 to 2012.Based on reviewing full text,the data were selected,evaluated and accessed.RevMan 5.1 and Stata 12.0 were used to perform the statistical analysis of those studies that were in accordance with the inclusive criteria.
According to the different ethnicities,the objects were divided into two subgroups as European and Asian to analyze respectively.Also,depending on different
 inheritances,the  objects were  divided into five patterns including C/T allele,CC/TT,CC/CT+TT,CC+CT/TT and CC+TT/CT genotypes to analyze  respectively,i
ncluding heterogeneity inspection,effect consoliating and publication bias assessment.Results A total of 11 studies were available for this analysis,in
cluding 2 443 schizophrenia patients and 2 469 controls.The Meta-analysis results showed that the allele of all people were OR=1.12，95%CI=0.96-1.31，P>0.05； CC/TT of all people were OR=1.11，95%CI=0.80-1.53，P>0.05；CC/CT+TT of all people were OR=1.13，95%CI=0.99-1.30，P>0.05；CC+CT/TT of all people were OR=1.18，95%CI=0.93-1.50，P>0.05；CC+TT/CT of all people were OR=0.95，95%CI=0.84-1.06，P>0.05.
Conclusion Current evidence is insufficient to show that HTR2A gene T102C locus polymorphism may be associated with schizophrenia,suggesting that the gene polymorphism has no significantly genetic association with schizophrenia.
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Objective To evaluate the relationship between smoking and p53 gene mutation in the lung cancer patients with Meta-analysis.
Methods PubMed,Web of Science,ProQest and Medline were used to search all the relevant studies about
 the association between smoking and p53 gene mutation in the patients with lung cancer.Based on reviewing full text,the studies were selected and evaluated and the data was extracted.Statistical analysis was performed with Stata 12.0 software including the heterogeneity inspection,publication bias assessment,sensitivity analysis,effect consolidating and cumulative Meta-analysis.Results Totally 15 articles with 1 770 lung cancer patients were identified. 69.6% of the patients were smokers,30.4% were non-smokers.Overall,the smokers with lung cancer had a 2.70-fold higher risk for p53 gene mutation than the non-smokers with lung cancer (OR=2.70,95%CI=2.04-3.59).Conclusion p53 gene mutation is associated with smoking in the patients with lung cancer.The smokers with lung cancer have higher risk for p53 mutation than non-smokers.

Objective To detect the levels of anti-gliadin IgA and anti-gliadin IgG antibodies in the plasma of the patients with schizophrenia,and to investigate the   association between schizophrenia and anti-gliadin IgA,IgG bodies in a Chinese Han  population,and to clarify the effect of gliadin on the occurrence of schiz
ophrenia.Methods The plasma samples were collected from 428 patients with schizophrenia and 555 cases of normal control subjects in a Chinese Ha
n population.The gliadin antibodies in plasma,including IgA and IgG,were tested using a native anti-gliadin ELISA test kit.The positive rates of plasma anti-gliadin IgA,and anti-gliadin IgG were tested by Chi-square test between the patients with schizophrenia and control subjects.The differences of the levels of
 anti-gliadin IgA and anti-gliadin IgG were tested by Mann - Whitney U test between the patients with schizophrenia and control subjects.
Results Compared with normal control group,the anti-gliadin IgA level and the positive rate of plasma anti-gliadin IgA in patient group were increased significantly(P<0.01),but there were no significant differences of the positive rate and the level of anti-gliadin IgG in plasma between patient and control groups (P>0.05).The anti-gliadin IgA levels in the patients with delusion of observation,delusion of being revealed,delusion of persecution,delusion of jealousy,delusion of grandeur,incoherence of thinking,illogic thought,bizarre behavior,aggressive behavior,hallucination-delusion syndrome,poverty of thought,emotional blunting / apathy and aboulia were higher than that of the normal controls ( P<0.05);the anti-gliadin IgG levels in the patients with delusion of being revealed and delusion of grandeur were higher than that of the normal controls ( P<0.05).Conclusion Gliadin is associated with the onset of schizophrenia  in  Chinese Han  population,and the plasma antibodies of gliadin maybe play an important rale in the onset of schizophrenia.

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Objective To investigate the association between CHI3L1 polymorphism and asthma in the Han population of Guangdong,and to explore the clinical values of chitinase-like protein YKL-40 in evaluating disease severity and monitoring the progression of the disease.Methods Total 251 objects were divided into asthma group(n=150) and control group(n=101).The single nucleotide polymorphism (SNP) at CHI3L1 gene was detected with Massarray method.The serum YKL-40 levels,total IgE levels,peripheral blood eosinophil percentage(Eos%) of the objects in asthma and control groups were detected.The asthma patients were divied into asthma exacergation group and stable group, and the serum YKL-40 levels were  compared.Results The distribution
of A/G at rs3806448 of CHI3L1 had  significant difference between asthma and control groups(P<0.01),and the frequency of A allele at rs3806448 of the pati
ents in asthma group was significantly higher than that in control group(OR= 1.93,95% CI=1.30－2.62,χ2=11.6,P<0.01).The serum YKL-40 levels，total IgE and Eos% of the patients with AA at rs3806448 were higher than those of the  patients with GG or AG at rs3806448 (P<0.05);the FEV1% of the patients with AA was lower than those of the patients with GG or AG at rs3806448(P<0.01).The serum YKL-40 level  of the patients in asthma group was significantly highe
r than that in normal control group(P<0.01),and the serum YKL-40 level of the patients in asthma exacergation group was higher than those in asthma stable group and normal control group(P<0.01).The serum YKL-40 level of the patients in asthma group was positively correlated with the peripheral blood Eos% and total IgE level(rEos=0.348，rIgE=0.437，P<0.01),and was negatively correlated with the lung function(r=－0.745，P<0.01). Conclusion
The polymorphism of CHI3L1-rs3806448 may correlate with asthma of Han population in Guangdong area.The serum YKL-40 may be a new biomarker to
 evaluate asthma disease severity and helpful for monitoring illness stage.

Objective To detect the percentages of CD4+,CD8+T lymphocytes in the breast invasive ductal carcinoma microenvironment, and to analyze their correlat
ion with breast carcinoma microvessel density (MVD).Methods The expressions of CD4,CD8 and CD34 were detected in 10 cases of adjacent normal breast tissue and 37 cases of breast invasive ductal carcinoma tissue by immunohistochemistry.The correlations between CD4,CD8,CD4/CD8 and MVD were analyzed by Spearman correlation analysis.Results The CD4 expression level (22.63%±11.53％) in breast invasive ductal carcinoma tissue was lower than that in adjacent tissue（34.09%±10.41％),and there was statistically significant difference (P<0.05).With the increasing of the histological grade of    breast cancer,,the expression level of CD4 was gradually decreased(P<0.05).The CD8 expression level (14.27%±9.03％) in breast invasive ductal carcinoma tissue was higher than that in adjacent tissue (10.51%±1.88％), but there was no statistically significant difference (P>0.05).With the increasing of the histological grade  of   breast cancer,the expression level of CD8 was gradually increased(P<0.01).The CD4/CD8 ratios in adjacent tissue group and breast invasi
ve ductal carcinoma tissue group were 3.24±1.02 and 2.16±2.17,and there was no statistically significant difference between them (P>0.05).
With the increasing of histological grade of breast invasive ductal carcinoma,the CD4/CD8 ratios were decreased (P<0.01).The MVD in breast invasive ductal carcinoma tissue was obviously higher than that in adjacent tissue,and it showed an increasing trend with the increasing of  the histological grade of breast invasive ductal carcinoma(P<0.05).The correlation analysis showed that the MVD  was negatively correlated with the CD4 expression（r=－1.000，P<
0.001） as well as CD4/CD8 ratio（r=－0.984，P<0.001）,and was positively correlated with the CD8  expression level
（r=0.984，P<0.001）.Conclusion The expression of CD4 is decreased in breast cancer tissue,the expression of CD8 is elevated,and the CD4/CD8 ratio is decreased,which suggests that T cells in the breast cancer microenvironment are immunosuppressed.
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Objective To identify normal breast tissue and breast invasive ductal carcinoma (IDC) tissue by shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS),and to explore the biological characteristics of IDC and the identification method by discussing its spectroscopy characteristics.
Methods The frozen section of breast tissue from 24 patients (female,aged 27-59 years) underwent routine surgical resection were obtained.9 cases of normal breast tissue and 15 cases of IDC breast tissue were detected by Raman spectroscopy and then SHINERS technique was utilized.A total of 263 Raman spectra and 249 SHINERS spectra were obtained.All the spectra were dealt with baseline corrected by fitting and subtracting a third-order polynomial and then smoothed with a 15-point Adjacent-Averaging.Results The characteristic peak of normal breast tissue appeared at 1 090,1 157,1 262,1 300,1 442,1 658,1 745,and 1 874 cm-1.After adding shell-isolated nanoparticles (SHINs),some peaks shifted at 2-3 cm-1,and the relative strengthes of 1 090 and 1 157 cm-1 were significantly increased,and the  characteristic peak at 1 496 cm-1 appeared.The Raman spectroscopy showed there were more nucleic acid ch
aracteristic peaks (including 878,1 086,1 157 cm-1) in the breast IDC tiss
ue;
after
adding SHINs,the relative strengthes of 1 004,1 157,1 526,and 1 658 cm-1 were  increased,the characteristic peak of lipid at 1 745 cm-1 and 1 442 cm-1
 stemmed from the C=O and CH2 stretching vibration.Compared with the normal breast tissue,the breat IDC tissue showed a blue shift of 2-3 cm-1.The characteristic peaks of nucleic acid had the blue shift from 1 086 cm-1 to 1 090 cm-1,and the characteristic peak of β-carotene emerged at 1 527 cm-1.
Conclusion Raman spectra can discover the differences of the characteristic peaks of DNA,β-carotene,and protein between breast IDC and normal breast tissues;SHINERS can distinguish breast IDC tissue from normal breast tissue successfully.

Objective To research the effect of the Kiss-1 gene promoter methylation on the Kiss-1 gene expression in colorectal carcinoma tissue,and to analyze
 the relationship between the Kiss-1 gene methylation and the clinical pathological features of colorectal carcinoma and its clinical significance.
Methods The Kiss-1 gene promotor region methylation,Kiss-1 gene mRNA and protein expressions  were detected respectively by methylation-specific PCR,Real-time PCR and Western blotting method in 126 cases of colorectal carcinoma tissue and para-cacinoma  normal  colorectal tissue.Results The positive rate of Kiss-1 gene methylation in colorectal carcinoma tissue (83.33%) was significantly higher than that in normal tissue (30.16%) (P<0.05).In the cancer tissue,the expression levels of Kiss-1 gene mRNA and protein in Kiss-1 gene promoter methylation positive group was lower than
 that in Kiss-1 gene promoter methylation negative group (P<0.05).The positive rate of Kiss-1 gene promoter methylation in T3+T4 stage group was higher than that in T1+T2 stage group (P<0.05),but the Kiss-1 gene mRNA and protein expression levels were lower than those in T1+T2 stage group (P<0.05).In the poorly and moderately differentiated group,the positive rate of Kiss-1 gene promoter methylation was higher than that in well differentiated group (P<0.05),but the mRNA expression level of Kiss-1 gene was lower than that  in well differentiated group (P<0.05).In lymph node metastasis
 group,the positive rate of Kiss-1 gene promoter methylation was higher than that in lymph node negative group (P<0.05),but the mRNA and protein expression levels were lower than those in lymph node negative group (P<0.05). And the positive rate of Kiss-1 gene promoter methylation in distant metastasis group was higher than that in non-distant metastasis group (P<0.05),but the mRNA and protein expressions levels  were lower than those in non-distant metastasis group (P<0.05).There were no significant associations between the positive rate of Kiss-1 gene promoter methylation and the  gender,age,tumor location,and tumor diameter of the patients (P>0.05).Conclusion The Kiss-1 gene promoter methylation in colorectal carcinoma  tissue is associated with the Kiss-1 gene expression level and the malignant characteristics of colorectal carcinoma;Kiss-1 gene promoter methylation may be used as a reference indicator for predicting the risk of metastasis  of colorectal carcinoma.
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Objective To study the incidence frequency of prothrombin G20210A (FⅡ G20210A) mutation in the patients with pulmonary thromboembolism(PTE) in northeast China,and to clarify thepredictive value of FⅡ G20210A mutation detection in PTE of the population in northeast China.
Methods 60  PTE patients(PTE group) and 80 sex-matched healthy controls(control group) from the same geographic area were selected.All
the patients were diagnosed by lung ventilation/perfusion scan and/or multi-slice CT pulmonary angiography(CTPA) as well as medical history.The genome DNA was extracted from the whole blood using alcohol.Polymerase chain reaction(PCR),restriction fragment length polymorphisms(RFLP)  analysis with HindⅢ restriction enzyme and sepharose gel electrophoresis were used to identify the FⅡ G20210A mutation in PTE group and control group.
Results After digested by HindⅢ restriction enzyme,only the fragments of  407 and 99 bp were found in PTE group.The frequency of
 FⅡ G20210A mutation was 0%,there was no statistical difference compared with contol group(P＞0.05).There were no heterozygote and homozygote mutation of FⅡ G202210A gene in PTE group and control group.Conclusion The incidence of FⅡ G20210A mutation in the PTE patients in northeast China is very low,and the detection  of  FⅡ  G20210A mutation may have no predictive  value in PTE of the population in northeast China.

Abstract:Objective To compare the dosimetric parameters of volumetric modulated arc therapy（VMAT）,fixed field intensity modulated radiation therapy（IMRT） and three-dimensional conformal radiotherapy（3D-CRT）in the radiotherapy for the patients with locally recurrent nasopharyngeal carcinoma,and to analyze their characteristics.Methods Twelve  patients with locally recurrent nasopharyngeal carcinoma  were treated with VMAT,IMRT and 3D-CRT plan designed by Pinnacle 9.2 and Preciseplan 2.03 treatment planning system.The dosimetric parameters of targeted volumes and organs at risk were compared between three groups.Results The conformation indexes(CI) of VMAT and IMRT plans were similar,and they were both better than 3D-CRT plan,the difference  was significant（P<0.05）.The homogeneity index(HI) in three groups were similar,there were no statistically significant differences between them(P>0.05).The monitor units(MU) and beam time in 3D-CRT group were bettethose in other two groups,and VMRT group was better than IMRT group,the statistical differences were observed between three groups（P<0.05）.There were no statistical differences of organs at risk such as brainstem and lens between three groups（P>0.05）.The doses of the spinal cord,optic nerve,optic chiasm and temporal lobe of brain in VMAT and IMRT groups were better than those in 3D-CRT group,there were statistical differences between them（P<0.05）,and the data in VMAT and IMRT groups were similar,and there were no statistical differences（P>0.05）.Conclusion There are differences of the targeted dose distribution between the three kinds of radiation technology,while VMAT and IMRT plans  can cover the targeted areas and reduce the received doses of organs at risk.The CI,MU and beam time of VMAT plan are better than those of IMRT plan.3D-CRT plan only has advantage in the MU and beam time.

Objective To observe the therapeutic effect of Tanreqing Injection in treatment of acute irradiation oropharyngeal mucositis in the patients with head and neck cancer, and to provide reference for its clinical application．Methods Fifty patients with head and neck cancer were randomly divided into treatment group and control group(n=25)．The patients in treatment group received concurrently intravenous infusion of Tanreqing Injection during irradiation course. The patients in control group received daily regular oral nursing；oropharyngeal mucositis was evaluated according to the protocol proposed by RTOG. Results  The patients in  treatment group with Ⅰ and  Ⅱ oropharyngeal mucosal injury accounted for 76%,and the patients in  control group with Ⅲ, Ⅳ oropharyngeal mucosal  injury accounted  for 60%, there was  statistically significant difference between two groups (P<0.05).The scores of the  patients with oropharyngeal pain in treatment group  were 0-3 (mild), in control group they were  4- 6 (moderate).The  patients with mild,moderate,and severe oropharyngeal pain in treatment group and control group  after radiotherapy accounted for 72％,24％,4％ and 12％,64％,24％;there were statistically significant differences between two groups (P<0.05).Conclusion Tanreqing Injection has a good therapeutic effect on acute radiation stomatitis and  layngopharyngitis，and it has the effects of relieving oral and pharyngeal mucosa injury, anti-inflammatory and analgesic and accelerating wound healing.

Objective To explore the influence  of 8-week aerobic exercise in heart rate variability (HRV) in obese adolescents and to provide the evidence for form ulating special exercise prescription.Methods Forty male obese adolescents (BMI≥25 kg。m^-2) were randomly divided into exercise group (n=20) and control group (n=20).The  subjects in exercise group conducted a 8-week (five times/week,50-60 min) aerobic exercise while those in control group maintained their routine lifestyle.The body composition,hemodynamic parameters including heart rate(HR)  and blood pressure and HRV parameters including total power (TP),normalized low frequency (LFn),normalized high frequency (HFn) and LFn/HFn ratio were measured before and after experiment.Results There were no significant differences of the indicators of the subjects  between two groups before experiment (P>0.05).At the end of experiment,compared before experiment,the fat mass(FM),the  percentage of body fat(BF%),the rest HR,SBP and DBP were decreased (P<0.05 or P<0.01);the  TP  and HFn were increased(P<0.01);the LFn  and LFn/HFn ratio were  reduced (P<0.01) in exercise group after experiment;but the indexes of the subjects in control group showed no significant changes (P>0.05).Compared control group,the  FM,the BF%,the rest HR,SBP,DBP,LFn  and LFn/HFn ratio were decreased  while the TP  and HFn  were increased (P<0.01)  in exercise group after experiment.Conclusion Aerobic exercise may effectively reduce the  sympathetic activity and increase the  vagal tone in the  obese adolescents.

Objective To compare the differences of pulmonary nodule detection rates between iterative reconstruction (sinogram affirmed iterative reconstruction,SAFIRE) algorithm and filtered back projection （FBP）algorithm in chest CT,and to evaluate the detection accuracy.Methods Three groups of tube voltage values of 80,100,and 120 kV were defaulted on the new dual-source CT，with autom
aticmAs care dose 4D technology,the chest phantom with simulated pulmonary nodules was scanned,then the images were reconstructed with FBP and SAFIRE (grade 1-5),respectively.The  detection rates of simulated pulmonary nodules  in the chest CT images reconstructed of SAFIRE (grade 1-5) and FBP were compared，and their diameters and CT values were measured.Results
With the same tube voltage,no significant difference was found in the detection rate of simulated pulmonary nodules between SAFIRE (grade 1-5) and FBP (P>0.05),the diameter deviation of simulated nodules of SAFIRE (grade 3) was less than FBP，and the difference in the average CT value of the simulated nodules between SAFIRE (grade 3) and FBP was not statistically significant (P>0.05);the simulated nodule detection rate of 100 kV was equivalent to the detection rate of 120 kV,the simulated nodule （－800 HU and 3 mm ）detection rate of 80 kV was less than that of 120 kV;as the tube voltage reduced,or simulated nodule diameter decreased，or the density of simulated nodule reduced,the nodule’s diameter deviation was increased.Conclusion Compared with FBP,the capabilities of SAFIRE in pulmonary nodule detection in different densities and different sizes are same,and SAFIRE algorithm is helpful for accurate displaying of pulmonary nodules,and it can be used for low-dose CT lung cancer screening program.

Objective To study the application value of microvascular density(MVD) in determination of transrectal ultrasound hemodgynamics in the differential diagnosis of prostate cancer and chronic prostatitis,and to provide imageological basis for their differential diagnosis.Methods A total of 61 patients suspected of  prostate cancer underwent transrectal ultrasound scan and ultra
sound guided biopsy.38 cases of prostate cancer and 23 cases of chronic prostatitis were confirmed by pathology and retrospectively analyzed.The peak systolic blood flow velocity (Vs)  and blood flow classification of the  suspicious lesions were compared and analyzed.The MVD was observed by Weidner method with monoclonal antibody CD34 immunohisochemistry staning.Results The Vs and the blood flow classification of the suspicious lesions in  prostate cancer group were significant higher than those in chronic prostatitis group  (P<0.05).The MVD in prostate cancer group and chronic prostatitis group were 46.70±13.87 and 34.38±7.28,respectively(P<0.05);the MVD in prostate cancer (C+D)stage and (A+B)stage were 56.99±12.85 and 39.97±10.21,respectively(P<0.05);the MVD in prostate cancer with high Gleason score group and low Gleason score group were 53.79±13.30 and 36.96±7.24,respectively(P<0.05).The Vs and the blood flow classification of the suspicious lesions of prostate cancer had a significantly positive correlation with the MVD　(r=0.793,P<0.05;r=0.723,P<0.05).Conclusion The Vs and the blood flowclassification of prostate cancer by ultrasound can well reflect the changes of the micrangium in tumor tissue.The  Vs and blood supply grade of suspicious lesions in the patients with prostate cancer are significantly higher than those in the patients with chronic prostatitis.They may be useful for the identification of prostate cancer and chronic prostatitis.

Abstract:Objective
To understand  the prevalence of hypertension and selenium intake of the residents in rural areas of Jilin province,and to discuss the relationship between selenium intake and the prevalence of hypertension, and to provide reference for prevention and therapy of  hypertension.Methods A total of 1 380 inhabitants in rural areas of Jilin province were selected by way of cluster sampling to conduct dietary investigation,physical and blood examination.The selenium intake was categorized into three groups according to tritiles after adjusted by energy,and the relative risk for incidence of hypertension in each group was estimate by using the Logistic regression model with the first group as the reference.Results The prevalence ofhypertension in the study area was 37.4%. The average for selenium intake was 35.908 μg?d-1 and was lower than the standard level of selenium recommended by  RNI.After adjustment for other risk factors,the relative risk for incidence of hypertension for different selenium intakes in the  men was 1.00,0.471（95%CI 0.290－0.765）,and 0.596（95%CI 0.373－0.951）,the P values were 0.002 and 0.030.There was no significant relationship between selenium intake and the incidence of hypertension in the  women,and there was also no significant relationship between Han people and Korean people in China.Conclusion The selenium intake is the influencing factor affecting the incidence of hypertension of the male rural residents in Jilin province,which reminds that the increasing selenium intake is helpful for the prevention of male hypertension.

Abstract:Objective
To establish a high-performance induction culture system for Raw264.7 cells to differentiate into osteoclasts（OC）　in vitro　 by improving the cell culture program.Methods The Raw264.7 cells were cultured with α-MEM medium containing 50 μg•L^-1 M-CSF,100 μg•L^-1 RANKL,and 1×10-8  mol•L-1  1α,25-（OH）2D₃  in 5% CO₂ for 12 d at 37℃. The cells were digested transiently every time before the medium was changed after every three days.The morphologic changes of the Raw264.7 cells were observed by  inverted microscope.The maturation and phagotrophic function of OC were identified by HE,TRAP,FITC-phalloidin staining and immunofluorescence.Results The cells remained to grow in single layers all the time in most areas of the well during the whole induction by the improved culture program.The observation results of inverted microscope and HE staining showed that the growth area of the polykaryotic OC reached to 70% of the well on day 12. FITC-phalloidin staining showed that in the maturation of the OC,the cluster-shaped podosomes in the pseudopodia gradually transformed into rings,which finally fused to form a large belt surrounding the periphery of the cytoplasm.The calcitionin receptor (CTR)  expressed by OC was markedly enhanced compared with the precursor cells by  immunofluroescence staining,and a large number of red granules appeared in the cytoplasm of OC with TRAP staining on day 12.  These results comfirmed that the obtained  OC were maturated and owned phagotrophic function.  Conclusion A high-performance induction culture system for Raw264.7 cells to differentiate into OC in vitro induced by combination of 50 μg•L^-1 M-CSF,100 μg•L^-1 RANKL, and 1×10^-8  mol•L^-1 1α,25-(OH)2D3 is established by improving the cell culture program.