Abstract:Objective To study the expressions of adenosine and serotonin (5-HT)-related genes in the preoptic anterior hypothalamus (PO/AH) of rats with habenular lesions and with zolpidem treatment and to explore the effect of the habenular nucleus in the sleep-promoting regulation of the PO/AH.Methods Both a rat model with habenular lesion and a hypnotic-treated rat model were established by passing a DC current or by infusing with zolpidem into the stomach and then the rats were divided randomly into habenular sham group,habenular lesion group,vehicle group and zolpidem group.The mRNA levels of adenosine and 5-HT-related genes were examined by RT-PCR in the PO/AH.Results Compared with sham group,only the mRNA expression level of adenosine kinase in the PO/AH had a little increase in habenular lesion group (P> 0.05).However,the expressions of adenosine A1 receptor and 5-HT-related genes (tryptophan hydroxylase and 5-HT2A receptor) were significantly up-regulated in the PO/AH of habenular lesion group (P< 0.05).Compared with vehicle group,the mRNA expression levels of 5-HT-related genes were obviously diminished in the PO/AH of zolpidem-treated rats. Adenosine kinase expression was not statistically decreased (P> 0.05),but adenosine A1 receptor expression just had slight up-regulation in zolpidem group (P> 0.05).Conclusion Habenular nucleus has a regulatory effect on the sleep-promoting PO/AH and its effect is performed by the pathways of adenosine and 5-HT in this process.

Abstract:Objective To investigate the expressions of MDR1 and P-glucose protein (P-gp) in brain tumor stem cells(BTSCs),and find new therapeutic targets and better anticancer strategies.Methods Tumor samples were obtained from consenting patients,actutely dissociated,triturated into single cells,and then inoculated into serum-free medium.After the primary brain tumor spheres generated,they were triturated again and passaged into fresh medium.Finally,they were digested into single cells with dispase.The specific marker CD133 was detected with immunofluorescent method.The expression of MDR1 in BTSCs was determined with immunohistochemical method,and the expression of P-gp was detected with Western blotting.Results The BTSCs isolated from glioblastoma,had the ability to propagate and differationate in vitro.The expression of CD133 was proved by immunocyt ochemistry and immunofluorescent method,the same with the MDR1,and the expression rate of MDR1 in the BTSCs was (45.3±5.4)%.P-gp expressed in BTSCs detected by Western blotting.Conclusion There are surely MDR1 and P-gp  expressions in BTSCs,which may make them more resistant to chemical therapy.

Abstract:Objective To explore the mechanism underlying Prohibitin 2 (PHB2)-mediated repression on myocyte enhancer factor 2 (MEF2) and facilitate the study of the role of PHB2 in other MEF2 positive cells. Methods Using lipofectamin transfection method, Hela cells were co-transfected with plasmid expressing PHB2 and MEF2 and luciferase reporter plasmid. 36 h after transfection, the recombinant proteins were tested with Western blotting and the activity of luciferase was measured with luminator in the supernatant of cell lysis.  Results Compared with the cells without PHB2 transfection, the inhibitory fold of co-transfected PHB2 on MEF2-dependent expression of luciferase was 0.28(P<0.01) and proportional to the amount of PHB2 expression. The activation fold of MEF2 transactivation domain on the expression of luciferase was 2.24±0.21 when co-transfected with PHB2, while it was 9.53±1.06 without PHB2 (P<0.01). Upon the treatment of TSA, the inhibitor for histone deacetylase (HDAC), the inhibitory fold of co-transfected PHB2 on MEF2 was changed from 0.38 to 0.98(P<0.01). Conclusion PHB2 represses the transcriptional activity of MEF2 by specifically acting on its transctivation domain and through HDAC to mediate the repression, but independently on MyoD and DNA binding activity of MEF2.

Abstract:Objective To investigate the hormesis effect on human marrow mesenchymal stem cells(MSCs)induced by low dose radiation(LDR) and its related signal transduction mechanism.Methods Human marrow MSCs and K562 cells were divided into control group,radiation group and SB203580 group.The dose of radiation was 75 mGy in three group.The expression levels of P38MAPK,P53,P21 and proliferation index(PI) were observed at 24 h after radiation,The expressions of  phosphor-P38MAPK(p-P38MAPK) of K562 cells and MSCs were observed immediatly,4,12 and 24 h after radiation.SB203580 was adjusted to 5 μmol·L-1 end  concentration according to the culture liquid volume in SB203580 group,and was added at 1 h before  experiment.Results The expression of p-P38MAPK of human  MSCs attained peak at 12 h after 75 mGy radiation,the expression levels of P53 and P21 protein were dereased,PI was increased,P38MAPK didn’t change;P53 and P21were increased,PI was decreased after P38MAPK kinase activity was inhibited  with SB203580;the expression levels of p-P38MAPK，P53，P21,P38MAPK and PI of K562 cells didn’t change at 24 h after 75 mGy radiation;PI was slightly decreased,P21 was slightly increased after addition of SB203580.Conclusion LDR can induce hormesis of human MSCs,the proliferation hormesis is mediated by P38MAPK signal and related to down-regulation of P21 which is P53 dependent.

Abstract:Objective To establish a method to screen a mouse reactive suppressive oligodeoxynucleotide (ODN) in vitro for future research in mice. Methods     Mouse splenocytes were divided into  control and  CpG 2216 groups. The supernatants of splenocytes induced by CpG 2216 with different concentrations(0,0.25,0.50,1.00，2.00，4.00，8.00，16.00 and 32.00 mg·L^-1) for different incubation time (3,6,12,24,48,72 and 96 h)were collected and detected by VSV protection bioassay to confirm the optimal concentration and incubation time of CpG 2216 and the dilution of the supernatant. Then the mouse splenocytes were divided into control group, CpG 2216 group, CpG 2216 + different concentrations of suppressive ODN(A151) group to find an optimal concentration of A151. At last,the  mouse splenocytes were divided into  control group, CpG 2216 group, CpG 2216 + suppressive ODN(developed by our lab) group. The anti-viral activities of the supernatants were detected by VSV protection bioassay using above conditions to practice the screening method established. Results  The screening method established was as followed: mouse splenocytes(cell density 5×10⁶·mL^-1) were cultivated with 2 mg·L^-1 CpG 2216 and/or 16 mg·L^-1 suppressive ODN for 24 h. The supernatants were collected and added to L929 cells by 1∶4 dilution. After 18 h incubation, the cells were cultivated  with  VSV for 48 h and A578 was detected to screen suppressive ODN. Conclusion Using CpG 2216 as the stimulator and A151 as the positive suppressive ODN, the conditions of screening experiment are established successfully and could be used for further development of novel suppressive ODN in mice.

Abstract:Objective To observe the expression of Humtingtin interacting protein 1(HIP1) in the injury of Hela cells induced by vitamin K3(VK3).Methods Hela cells in log phase were randomly divided into control group,VK3 group,VK3+NAC group and NAC group,respectively.The survival rate of Hela cells was determined by MTT.The expressions of HIP1 and NF-κB protein  in Hela cells were detected by Western blotting.The expression of Cyclin D1 mRNA  was tested by RT-PCR assay.Results Compared with control group,the survival rate of Hela cells in VK3 group decreased（P<0.05）;the  expressions of HIP1 and NF-κB protein and Cyclin D1 mRNA  were all decreased in VK3 group （P<0.05）.Compared with VK3 group,the survival rate of Hela cells in  VK3 + NAC group was increased（P<0.05）;the expressions of HIP1 and NF-κB protein  and Cyclin D1 mRNA  were increased（P<0.05）.Conclusion VK3 has an effect of inhibiting the cell proliferation which may be related with down-regulation of HIP1 protein expression.

Abstract:Objective To investigate the protective effect of Gross Saponin Tribulus Terrestris (GSTT) on the rat myocardial ischemia-reperfusion models and its mechanism.Methods Thirty-six male Wistar rats were divided randomly into sham group,ischemia-reperfusion model group,Xue-Sai-Tong(20　mg·kg-1) group and GSTT (30.0，15.0，7.50 mg·kg-1) groups(n=6).The myocardial ischemia-reperfusion model was induced by 30 min left anterior descending coronary occulusion and 120 min reperfusion in openchest anesthetized rats.The cardiocyte morphological changes were  observed and the contents of SOD,MDA,CK,AST,LDH and NO in serum were detected.Results Compared with sham group,the cardiocytes had serious edema and cytoplasm was stained slightly and nucleus were contracted and strained deeply after reperfusion in model group.In GSTT (30 and 15 mg·kg-1) groups and Xue-Sai-Tong group the  cellular edema was reduced,the cadiocytes maintained normal  appearance.Compared with sham group,the contents of AST,LDH,CK and MDA in model group were increased but SOD and NO were decreased（P<0.01）.Compared with model group,in GSTT（30 and 15 mg·kg-1）　groups and Xue-Sai-Tong group  the contents of MDA,AST,LDH and CK were decreased,and  the contents of SOD and NO were increased（P<0.05 or P<0.01）.Conclusion GSTT has the protective effects on myocardial ischemia-reperfusion injury in rats,which may be related to inhibition of lipid oxidation and modulation of endogenous antioxidant enzyme activities.

Abstract:Objective To study the inhibitory effects of triethyltin chloride (TETC) on proliferation and the morphological changes of rat C6 glioma cells in vivo.Methods C6 glioma cells were autotransplanted into 16 Wistar rats,and the rats were divided into experiment group and control group.TETC was injected into each rat through intraperitoneal route at the dose of 1 mg·kg^-1·d^-1 and the injection lasted 4 d in experiment group,and the physiological saline was injected into each rat in the same way as experiment group in control group.The weights of C6 glioma were measured and the proliferation rate of the tumor was calculated after 14 d.The morphological changes of C6 glioma cells were observed by HE staining and electron microscope.Results The weight of C6 glioma in TETC group was lighter than that in control group(P<0.01),and the proliferation rate was decreased compared with control group (P<0.01).The cell number and cell density in TETC group were lower than those in control group observed by HE staining and the chromatin margination could be observed  in  TETC group.Conclusion TETC can inhibit the proliferation of rat C6 glioma cells in vivo and the mechanism of which may be involved in apoptosis.

Abstract:Objective To evaluate the biocompatibility of chemically extracted acellular muscle grafts (CEAM) and to demonstrate its advantages as tissue engineering scaffold after transplantation into the spinal cords of adult rats.Methods Thirty-six male rats were randomly assigned to control group and CEAM group, and underwent spinal cord lateral hemisection.After preparation of successful models,treatment consisted of application of CEAM into the lesion gap,which was left empty in control  group.Six rats in each group were killed 1,2 and 4 weeks after induction of the injury.Sections were stained for quantification of microglia/macrophages using ED-1 on week 1,week 2 and week 4.Additionally,Holmes’ silver staining method was chosen to detect axonal regeneration,glial fibrillary acidic protein (GFAP) staining for detection of astrocytes in glial scars,and alkaline phosphatase staining for vascularisation on day 28. Results In control group,no definite axonal outgrowth into the lesion was found.The inflammatory response was most pronounced on day 7 and a dense non-oriented accumulation of astrocytes could be found at the edge of the lesion cavity in control group.In CEAM group,the number of regenerating axons in the scaffolds was 613.17±154.96,and they grew into the grafts in a strikingly organized fashion. The change of inflammatory response in CEAM group was the same as that in control group and there was no sign of the foreign body reaction induced by CEAM.In rats of CEAM group the astrocytes grew into the graft in a diffuse linear array.Vascularisation of the grafts was also confirmed.Conclusion CEAM is biocompatible with spinal cord,and it indicates that CEAM is a kind of good tissue engineering scaffold for the repair of spinal cord injury.
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Abstract:Objective To identify the major outer membrane proteins(OMPs) of Brucella Melitensis  with proteome analysis. Methods The OMPs were extracted from the culture of Brucella Ｍelitensis and then the two dimension electrophoresis was used to separate the OMPs.By means of matrix-assisted laser descorption/ionization time of light mass spectrometry (MALDI-TOF-MS),the OMPs were identified.Peptide mass fingerprints were searched in the NCBInr database using the Program Mascot.Results Sixty seven protein spots were identified,in which Omp25 and Omp31 were identified as two main OMPs and other OMPs,such as Imp,Frp and GroEL were found.The functions of identified OMPs  included thymidylate kinase,protein precursor,NADH dehydrogenase subunit,cytosol aminopeptidase,etc.Conclusion Identification and analysis of Omp25 and Omp31 provides an important clue for understanding the  molecular mechanism of   Brucella Melitensis and developing the vaccine to prevent
 Brucella attack.

Abstract:Objective To observe the expression of matrix metalloproteinase-2 (MMP-2) in U2OS cells transfected with human telomerase reverse transcriptase (hTERT),and explore the dependability between and  elomerase,and study the effect of telomerase activation in tumor infiltration and metastasis and their possible mechanisms of action. Methods  Recombinant plasmid PCI-Neo-hTERT  was  transfected into human osteosarcoma cell line U2OS.Two transfected positive clonings were chosen for experiment,while normal U2OS cells were used as control group.The effect of hTERT transfected into U2OS cells was detected by RT-PCR and TRAP-ELISA assays,zymography and RT-PCR were used to observe the expression and activity of MMP-2.Results Compared with control group,two positive clonings ectopic generated hTERT mRNA and expressed telomerase activity successfully(ΔA>0.2 U),while the MMP-2 mRNA without obvious difference(P>0.05),and the expression of proMMP-2  was decreased obviously(P<0.01).Conclusion Human osteosarcoma cell line U2OS transfected with hTERT could generate telomerase activity,moreover，the appearance of telomerase activity could regulate the secretion  of MMP to affect the degradation and construction of ECM which can affect tumor infiltration and metastasis.

Abstract:Objective To construct eukaryotic expressing vector of PTEN gene and to express the gene in the Hep-2 cell line and to clarify theraputic effect  of PTEN gene in the Hep-2 cell line,and  may laid foundation for further  study of PTEN gene treatment of laryngeal carcinoma.Methods PTEN gene was amplified from human normal laryngeal mucosa by RT-PCR and the fragment of the cDNA was cloned into the PTEN-Teasy vector.After the selection of the positive clone by DNA sequencing and restriction enzyme analysis,the fragment of the cDNA was cloned into the eukaryotic expressing vector of pEGFG-C1.The recombinant plasmid transfected into Hep-2 cell line and the gene transient expression was observed.with Western blotting.Results  A  1 400 bp DNA fragment was amplified with RT-PCR.The sequence analysis showed it was consistant with the sequence of PTEN gene in GenBank.Western blotting analysis provided strong evidence that PTEN gene was expressed successfully in transfected Hep-2 cell line.Conclusion The eukaryotic expressing vector of PTEN gene is constructed and PTEN protein is induced to express successfully.

Abstract:Objective To explore the role of PTEN in the development of oral squamous cell carcinoma (OSCC) by investigating the effect of PTEN on the proliferation of oral squamous carcinoma cells and its expression in oral leukoplakia(OLK) and OSCC.Methods The eukaryotic expression vector pcDNA-PTEN was constructed successfully,and then was transfected into squamous carcinoma cell line Tca8113.After the stablely transfected Tca8113 cell lines were established,MTT and flow cytometry were used to detect the growth and apoptosis of Tca8113 cells,respectively.The endogenous protein expression level of PTEN was determined by immunohistochemical method in 25 cases of oral leukoplakia and 32 cases of OSCC.0Results The over-expression of PTEN gene in pcDNA-PTEN group obviously inhibited the growth and induced the apoptosis of Tca8113 cells compared with  control group (P<0.01).Moreover,compared with control group,the cells at G0/G1 phase were significantly increased (P<0.05),but decreased at S phase (P<0.05).In OSCC,the expression of PTEN demonstrated absent or less than normal tissues and the expression level of PTEN had a significantly positive correlation with the malignancy of OSCC(P<0.05).Conclusion As a tumor suppressor gene,the expression of PTEN could suppress the growth of OSCC cell line in vitro. PTEN might play an important role in the development of OSCC.

Abstract:Objective To improve the administration method of lentiviral vector in rat CA1 region of hippocampus by electrode pipe installation and detect its validity and security.Methods Fourty-five male healthy SD rats were randomly divided into three groups: sham operation group(SH),normal saline group (NS),GFP gene lentiviral vector injection group(eGFP).The CA1 region was located by stereotaxic instruments and the lentiviral vector was administered by electrode pipe installation.At day 7,14 and 30 after the injection,SD rats were frozen sectioned and the expression of the green fluorescent protein was detected by fluorescence microscope.Nissl staining was used to detect the hippocampus injury.Results The stable expression of the green fluorescent protein was detected in rat CA1 region of hippocampus at day 7,14 and 30 using this method and the number of survival neurons had no significant difference between three groups (P>0.05).Conclusion The method of electrode pipe installation to inject lentiviral vector in rat hippocampus is successfully established and the high level expression of fluorescent protein can be detected with no significant injury,which lays the basis for its application in the gene therapy of the neurodamaged diseases.

Abstract:Objective To observe the expression of basic fibroblast growth factor(bFGF) at different stages of tooth germ development,and explore its mechanism and significance.Methods Immunohistochemical technique was used to examine the expressions of bFGF in the tooth germs at different stages of human fetus in 8-34 weeks,the suckling mice with 1-22 d,and the dog postnatal in 6 weeks-5 months.Results bFGF expressed in tooth germ at all stages.In human fetus,the positive reaction was observed in epithelium of tooth bud，dental plate,enamel organ,external and outer enamel. The ameloblasts,odontoblasts,epithelial root sheath and dental follicle showed strongly positive reaction in the unerupted human deciduous tooth,the tooth under the eruption of neonate external enamel,stratum intermedium cells,ameloblasts and odontoblasts showed strongly positive epithelium,dental papilla and dental follicle at bud stage and cap stage,while at bell stage cells in mice and 6 week postnatal canal tooth.The positive expressions of bFGF were observed in root of tooth,peridental ligament and the osteoblasts adjoin to peridental ligament in the dog succession of decideous teeth.Conclusion bFGF is one of critical regulator in the tooth development,which can regulate the differentiation and morphogenesis of tooth germ cells and control the eruption and succession of deciduous teeth at different stages.

Abstract:Objective To measure the level of interleukin-15 (IL-15) in serum and its expression in lung tissues，and analyze the correlations between IL-15 and IL-4，IFN-γ，eosinophil(Eos)，and explore the effect of IL-15 on bronchial asthma.Methods  Thirty femal BALB/C mice were randomly divided into three group (n=10),group A (asthma model),group B (corticosteroid treatment) and group C (normal control).All mice were killed 24　h after final OVA challenge.Blood were obtained for measurement of serum IgE，IL-4，IFN-γ and IL-15 levels by enzyme-linked immunoabsorbent assay (ELISA).Bronchoalveolar lavage fluid (BLAF) was collected for  Eos  count.The left lungs were isolated for pathological examination.The lung sections were stained with hematoxylin and eosin (HE).The expressions of IL-15 in lung tissues were measured by immunohistochemical SP method.Results ① The mouse asthma model appeared ethological changes  specific to asthma，the Eos count in BALF was increased，and IgE and IL-4 levels in serum were also increased compared with control group(P<0.01）.The lung tissues also appeared typical pathological changes and the inflammation cell score elevated.The level of serum IFN-γ was decreased（P<0.01）.Correlation analysis showed negative relationship between IL-4 and IFN-γ（r=－0.859，P<0.01），there existed Th1／Th2 imbalance.② The level of serum IL-15 in group A (77.97 ng·L-1±2.75 ng·L-1) was higher than that in group C (34.63 ng·L-1±1.44 ng·L-1)（P<0.01）.There were positive correlations between the  serum IL-15 and IgE，IL-4 levels(r= 0.681，P<0.01；r= 0.738，P<0.01），while the  serum IL-15 level had no correlation with IFN-γ. ③ The expression of IL-15 in lung tissues in group A was higher than that in group C (P<0.01）.There was a positive correlation between the expression of IL-15 in lung tissues and the Eos count in BALF (r= 0.787，P<0.01）.④ In mice treated with inhaled budesonide (group B），the Eos count in BALF ，the level of serum IgE ，the level of serum IL-15  and the expression of IL-15 in lung tissues were lower than those in group A（all P<0.01）.Conclusion IL-15 might contribute to promoting Th2 cells to secret cytokine IL-4，while not contribute to promoting Th1 cells to secret cytokine IFN-γ.IL-15 might be involved in the elevation of Eos and IgE，which suggests that IL-15 might be involved in the chronic inflammation of bronchial asthma.The inflammation of bronchial asthma could be inhibited by inhaled glucocorticoid.

Abstract:Objective To explore the effect  of exogenous purine nucleotide on the catabolism of nucleotide in heroin dependent rats,and clarify the mechanism of purine nucleotide treatment for heroin dependent rats. Methods  Fifty  male Wistar rats were randomly divided into control group,heroin administration group,heroin plus AMP and GMP group,heroin plus AMP group and heroin plus GMP group. The contents of uric acid(UA)，blood urea nitrogen (BUN) and creatinine (Cre) in plasma were measured,the activities of adenosine deaminase (ADA) and xanthine oxidase(XO) in plasma and brain cortex were measured by  test Kits.Results  The content  of UA in plasma and the activities of ADA,XO in heroin group were significantly higher than those  in control group and  three heroin plus purine nucleotides groups (P<0.05). The contents of BUN and Cre had no difference between various groups (P>0.05).The ADA and XO activities in plasma and brain cortex in heroin group were significantly increased compared with  control group and  three heroin plus purine nucleotides groups (P<0.05,P< 0.01 ).Conclusion Compensating purine nucleotides could ameliorate the enhancement of purine nucleotide catabolism.

Abstract:Objective To study the inhibitory effect of mistletoe alkali on the growth and invasiveness of human osteosarcoma cells (U2OS) in vitro. Methods The effects of mistletoe alkali on the growth of U2OS cells were detected by CCK-8 kit. The U2OS cells treated by 5-FU were used as positive control group and the U2OS cells without any treatment as  negative control group, the U2OS cells treated by mistletoe alkali were used as experimental groups. Meanwhile the growth of U2OS cells after treated by mistletoe alkali was assessed by cell growth curve. And the morphological changes of U2OS cells under different concentrations of medicine were observed. Furthermore, Their invasion ability was tested with Boyden chamber. Results Mistletoe alkali inhibited the growth of U2OS cells and the value of IC50 was 7 mg·L^-1. After treatment, the growth of U2OS cells slowed, and the cells became round, adherence decreased. Furthermore, the invasion ability of cells in the experiment group was obviously decreased. Conclusion Mistletoe alkali obviously inhibits the growth and the invasion ability of U2OS cells in vitro.

Abstract:Objective To explore the effect of donepezil hydrochloride on learning and memory function of normal   under age rats,and provide experimental  basis for  clinical application of donepezil hydrochloride.Methods Fourty Wistar rats just away from galactic were divided into control group(n=20) and donepezil hydrochlorid group(n=20),donepezil hydrochloride was given to the rats in donepezil hydrochloride group（0.45 mg·kg^-1，8 weeks）， the  effects of  donepezil hydrochloride on space discrimination ability and passive avoidance ability of normal  under age rats were detected with Morris water maze and Step-down   behavior test.Results Compared with control group,the latency,distance,average speed and starting angle of rats treated by donepezil hydrochloride did not change significantly  from the first day to the  sixth day，the time in platform， time in platform quadrant，times passing platform， distance in platform / total distance×100%  and average speed also did not change significantly in the seventh day  in the Morris water maze （P>0.05）; the error times of  rats treated by donepezil hydrochloride  did not decrease in the first day and in the second day,the latency did not prolong in the second day in Step-down  behavior test（P>0.05） .Conclusion Donepezil hydrochloride do not improve the  learning and memory function  of    normal under age rats.

Abstract:Objective To discuss the effects of ginsenoside Rb1 on bone lead content,learning and memory,the activities of superoxide dismutase (SOD) and nitric oxide synthase(NOS) in brain of  mice exposure to lead.
Methods The lead-exposed models of mice were induced by lead acetate contained in drinking water.The mice were divided into control group,model group and different doses of ginsenoside Rb1 groups （100,50,25 mg·kg^-1）.　Passive avoidance experiment and Morris water maze test were used to evaluate the effects of ginsenoside Rb1 on learning and memory of lead mice,and to determine the bone lead content,the activities of SOD and NOS in brain.Results Compared with control group,lead-exposuring could lead the following results: shortening the incubation period of avoid dark  experiment（P<0.01）,increasing the number of errors（P<0.05）,extending the search distance and the incubation period in Morris water maze test（P<0.05）,reducing the activity of SOD,increasing the activity of NOS and the bone lead content（P<0.01）.
Compared with model group,the incubation period of  passive avoidance experiment was obviously extended,the number of errors was clearly reduced,the search distance and the incubation period in ginsenoside Rb1 groups were apparently shortened（P<0.05 or P＜0.01）,the activity of SOD was increased（P<0.05 or P<0.01）,the activity of NOS and the bone lead content were decreased（P<0.05 or P<0.01.All the effects were dosage-dependent.And the effect of 25 mg·kg^-1 ginsenoside Rb1 was significantly different  compared with   100 and 50 mg·kg-1 ginsenoside Rb1（P<0.05）.Conclusion Ginsenoside Rb1　can reduce the lead content significantly,and obviously alleviate learning and memory of lead mice.It could meanwhile effectively resume the vitality of the antioxidant system of lead mice and protect their nerve cells.

Abstract:Objective To investigate the therapeutic effect of glycyrrhiza active substance on guinea pig model of vitiligo,and to clarify its mechanism on promoting synthesis of melanin,and to provide the basis for its clinical application.Methods Guinea pigs decolorized by H₂O₂ were used as experimental models,and normal control group was set up.After treatment of glycyrrhiza active substance in three doses groups(20,40,and 80mg·kg^-1),the therapeutic effect was observed by visual observation,macrography,histological examination as well as immunohistochemical study.Results The significant differences of skin color among control group,model group and treatment groups were observed by macrography.The total effective rates in 40 or 80 mg·kg^-1 groups were higher than that in control group (P<0.05 or P<0.01),and increased with the dose of glycyrrhiza active substance.Histological examination showed an evidently higher of melanin in basal cells,stratum spinosum and hair follicles in 40 or 80 mg·kg^-1 treatment groups compared with model group(P<0.01).Immunohistochemical results showed the expressions of HMB45 were higher in 40 or 80 mg·kg^-1 groups than that in model group (P<0.05 or P<0.01),and had dose-dependent relationship with glycyrrhiza active substance.Conclusion Glycyrrhiza active substance could increase the expression of HMB45 and promote synthesis of melanin,it shows certain efficacy in treatment of vitiligo,and it  can be used in treating vitiligo in clinic.

Abstract:Objective To observe the effects of mixture from ginsebosides and astragalus polysaccharides (MGA) on immune and antioxidative function in immunosuppressive mice induced with cyclophosphamide(CTX),and provide the experimental basis for clinical application of MGA.Methods Sixty Kunming mice were randomly divided into normal control group,model group and 100,200,400 mg·kg-1 MGA groups.The mice in MGA groups were administered by intraperitoneal injection for 15 d，The mice in normal control group and model group were administered with same volume physiological saline.On the 3rd and 10 th day,the mice in all groups except control group were intraperitoneally injected with CTX 100 mg·kg-1,same volume physiological saline in control group.On the 15th day,the weights of immune organs,phagocytic funtion of macrophage system,periphery leukocytes,the activitie of GSH-Px in whole blood and SOD in plasma,and the content of MDA in plasma were detected. Results As compared with the controls,the weights of thymus and spleen,phagocytic index and phagocytic activity in immunosuppressive mice were significantly decreased（P<0.05 or P<0.01）;the peripheral blood WBC count was decreased significantly (P<0.001 ).The activities of blood GSH-Px and plasma SOD were significantly decreased  (P<0.01 ),the plasma MDA content was markedly increased (P<0.05).It was indicated that CTX could cause immune suppression in mice and decrease anti-oxidation function.While compared with model group,the immune organ weights,phagocytic index,phagocytic activity and peripheral blood WBC count in 200 mg·kg-1 and 400 mg·kg-1 MGA  groups were increased (P<0.05 or P<0.01 ),the activity of blood GSH-Px was significantly increased (P<0.05 ),the plasma MDA content was markedly decreased （P<0.05).The weight of thymus and the plasma SOD in 400 mg·kg-1 MGA  group were significantly increased (P<0.05 or P<0.01).
Conclusion MGA has the effect of improving immune and antioxidative function in immunosuppressive mice induced with CTX.It indicates that MGA can be used as a medicine of assist treatment with chemotherapy of tumor.

Abstract:Objective To explore the effect of  triptolide (TL) combined with  hyperthermia on proliferation and apoptosis of  Hep-2 cells and its affect on cell cycle phases,and provide theoretical foundation for treatment of laryngocarcinoma.  Methods Hep-2 cells at logarithmic growth phase were randomly divided into control,hyperthermia,TL,TL　combined with  hyperthermia groups.MTT method was adopted to investigate the proliferation inhibition  rate  of the Hep-2 cells.Cell cycle and apoptotic rate of Hep-2 cells were analyzed by flow cytometry,and then  the changes of  subcellular structures were observed by electron microscope.
Results The proliferation inhibition rates in hyperthermia,TL,TL　combined with hyperthermia  groups were 21.2%,28.5% and 54.5%,respectively;the proliferation inhibition rate in TL combined with hyperthermia  group was higher than those in hyperthermia and TL groups(P<0.01).Compared with control group,the number of cells at G1 phase was increased (P<0.01),the number of cells at S phase was decreased(P<0.01),the number of cells at G2/M phase was relatively increased(P<0.01),the apoptotic rate was increased(P<0.01) in hyperthermia,TL,TL combined with hyperthermia groups;compared with hyperthemia and TL groups,the indexes mentioned above in TL combined with hyperthermia group had significant differences(P<0.01).The result of electron microscope indicated chromatin margination,mitochondrion swelling,apoptotic body formation in heperthermia,TL,TL combined with hyperthermia groups,no changes in control group.Conclusion TL combined with hyperthemia can　inhibit the proliferation of Hep-2 cells,induce apoptosis of Hep-2 cells and arrest the Hep-2 cells at G1 phase.

Abstract:Objective To explore the mechanism of modified amantadine（NAM）against influenza A(H5N1) virus and to provide experimental basis for development of NAM.  Methods DCK cells were cult ivated in vitro. Normal control group, virus control group and test group were included in this study. ① Cytopathic effect(CPE) with MTT was used to detect the inhibitory effect of NAM against influenza A(H5N1) virus. The 50% inhibitory concentration(IC50) and therapeutic index(TI) of NAM on influenza A(H5N1) virus were studied in three ways: adding NAM before contracting virus, contracting virus before adding NAM and contracting virus with adding NAM at the same time. ② The neuramidinase(NA) inhibitory effect was observed to obtain the effect of NAM on neuramidinase activity. Results ①Positive correlation was observed between logarithm of dose and protect rate of NAM in MDCK cells in the three different ways(0.95，0.95 and 0.99 ; all P<0.05). IC50 of NAM was 15.32 mg·L^-1 and TI was 103.31 when adding NAM before contracting virus, IC50 of NAM was 30.78 mg·L^-1 and TI was 28.10 when contracting virus before adding NAM, and IC50 of NAM was 203.92 mg·L^-1 and TI was 4.24 when contracting virus with adding NAM at the same time. ② NAM can inhibit neuramidinase activity, and IC50 was 87.36 mg·L^-1. Conclusion NAM can better inhibit influenza A(H5N1) virus in cells, and has some effects on interfering viral adsorption and invading the process of cell uncoating. NAM can also inhibit neuramidinase activity to some extent.

Abstract:Objective To establish rat models with pulmonary artery hypertension induced by monocrotaline(MCT) and to observe the pathological changes of lung tissue.Methods Seventy male Wistar rats were randomly devided into three group:control group(n=10),50 mg·kg^-1 MCT group(n=30),60 mg·kg^-1 MCT group (n=30).At two weeks and four weeks after injected intraperitoneally with MCT(injected intraperitoneally with equal normal saline in control group),the right ventricular systolic pressure(RVSP)and right ventricle weight/left ventricle+septum weight ［RV/(LV+S)］ ratio were measured.Hematoxylin -eosin staining and orcein technique were used to observe the pathological changes of lung tissue and pulmonary arterioles’medial thickness.Results Two weeks or four weeks after MCT administration,RVSP in 50 mg·kg^-1 MCT group was higher than that in control group（respectively 36.6 mmHg±5.1 mmHg，39.1 mmHg±7.0 mmHg versus 26.1 mmHg±3.8 mmHg，both P<0.01）,and  RV/(LV+VS) weight ratio increased greatly（respectively 0.289 4±0.021 7，0.409 4±0.125 6 versus 0.247 3±0.019 3,both P<0.01）,so were the results in 60 mg·kg^-1 MCT group compared with control group:RVSP respectively 33.8 mmHg±5.5 mmHg,46.6 mmHg±12.6 mmHg versus 26.1 mmHg±3.8 mmHg，both P<0.01;RV/(LV+VS) weight ratio,respectively 0.308 2±0.058 4,0.417 5±0.103 7 versus 0.247 3±0.019 3,both P<0.01.There was no significant difference of RVSP and RV/(LV+VS) weight ratio between two different doses of MCT groups. The pulmonary arterioles were markedly thickened in both 50 and 60 mg·kg-1 MCT groups compared with control group（21.3%±1.9%,23.4%±2.3% versus 11.3%±1.2%,respectively P<0.01).The rats in 60 mg·kg^-1 MCT group were half dead by the day 47 and none survived by the day 63,yet none of rats in 50 mg·kg^-1 MCT group was dead by the day 47 and seventy percent survived by the day 63.Conclusion Both 50 and 60 mg·kg^-1 MCT  can induce pulmonary artery hypertension and there were no pathological differences between them，but the survival of rats treated by the former dose is longer than that by the latter one.

To observe the repairing effect of marrow derived mesenchymal stem cells (MSCs) on hepatic fibrosis in rats，and provide a reliable theoretical basis for further clinical application of MSCs.Methods The MSCs of rats were isolated,purified and cultivated in vitro.The model of hepatic fibrosis induced by CCl4 was established.The rats were divided into three groups:normal control group,model group,MSCs transplantation group,6 each group.The MSCs were transplatated into the rats through tail vein in MSCs transplantation group.The repairing effect of MSCs on hepatic fibrosis was observed  and the expression of α-SMA in liver tissue was detected by immunohistochemistry  30 d after transplantation.Results  93.27% MSCs were at G0/G1 phase,and the CD44 surface expression in the MSCs was positive.The hepatic pathological injury changes of rats in MSCs transplantation group were significantly reduced than that in model group  30 d after MSCs injection.The lobular structure and liver sinusoids were normal and hyperplasia of fibrous connective tissue was not obvious.The intensity of α-SMA staining in MSCs transplantation group was lower than that in model group.Conclusion MSCs have  repairment on hepatic fibrosis in rats to a certain extent.

Abstract:Objective To establish the determination method of total polyphenols in vitis amurensis seed extract.Methods Using gallic acid as reference substance，the content of total polyphenols in Vitis amurensis seed extract was determined by the optimum conditions of Folin-Ciocalteu chromatometry.Results There was a good linearity between gallic acid concentration and absorbance in the range 0.002-0.010 g·L^-1（Y=86.243X+0.007,r=0.9996）；the precision RSD was 0.14%；the stability RSD was 1.77%； the repeatability RSD was 1.44%； the average recovery rate was  102.00%（RSD=1.37%）.Conclusion This method is simple，fast and has high sensitivity and good reproducibility.

Abstract:Objective To explore the method of obtaining and expanding epithelial stem cells (ESCs) from plucked hair follicle in vitro and clarify that fibroblasts are necessary for ESCs growth.Methods A amount of ESCs from plucked hair were obtained,the epithelial outgrowth of human plucked hair follicle was studied and compared between different culture conditions including no coating well,rat tail-collagen coating well,collagen Ⅳ coating well and 3T3 fibroblasts feeder.The characteristics of these cells were detected by RT-PCR and Immunofluorescent staining for CK15.Results After compared between different culture conditions including no coating well,rat tail-collagen coating well,cllagen Ⅳ coating well and 3T3 fibroblasts feeder,the results indicated that the plucked hair fragment B3 (bulge area of hair follicle) placing on 3T3 fibroblasts feeder was the optimum condition.RT-PCR and immunofluorescent method  confirmed that these cells expressed CK15 which was the special marker of ESCs.Conclusion ESCs can be obtained from bulge area of hair follicle which grows on the  3T3 fibroblasts feeder and this method is the optimum condition for ESCs growth after compared between different conditions.

Abstract:Objective To study the inhibitory effect of phenylacetate(PA) on hepatic carcinoma cells for developing of gene therapy. Methods SSMC-7721 cells  were treated with or without PA (0.25,0.50,1.00,2.00,4.00 mmol·L^-1) for 24,48 and 72 h,respectively. The cellular proliferation inhibitory rate was evaluated by MTT assay. Flow cytometry was used to estimate the changes of cell cycle. The morphous of tumor cells suppressed by PA was observed with electronic microscope. Finally,the activity of Caspase-3 protein  to estimate the way of apoptosis of SSMC-7721 cells under the control of PA was determined.Results The growth of SSMC-7721 cells were inhibited by 0.25-4. 00 mmol·L^-1 PA for 24-72 h.With the increase of concentration of PA, the proliferation inhibitory rate were 7.2%-73.1 % at 24 h, 9.1%-87.5% at 48 h, 15.8%-91.9% at 72 h.With the prolongation of  treating time and the increasing  of  PA concentration, the inhibitory rate was increased. 2.00 mmol·L-1 was the best concentration for the experiment. The cell cycle was arrested at G0 / G1 and G2/M phase. Some characteristic changes of apoptosis were observed  by electronic microscope,the activity of Caspase-3 protein in SSMC-7721 mediated by PA was increased.Conclusion PA could effectively inhibit SSMC-7721 cell proliferation and induce the apoptosis. Caspase-3 protein at least partly participate in the mechanism.

Abstract:Objective To elucidate the relationship between mandibular lateral displacement and mandible asymmetry by observing the morphological changes of condylar head and mandible during mandibular lateral displacement in growing rats. Methods Forty-eight male Wistar rats at the age of four weeks were divided at random into experimental and control groups, with 24 rats in each group. A super-hard resin plate was cemented to upper incisors to displace rat mandibles 2 mm to the left during closure (Ipsilateral side). A metal crown was fitted to lower incisors. The rats were killed 2, 4, 8 or 12 weeks after appliance attachment. The mandible was dissected out and halved. The length and width of condylar head were measured with a caliper. Radiographic films of the mandibles were exposed, and selected measurements were made.
Results The length of condylar cartilage on the ipsilateral side was significantly larger than contralateral side in experimental group(P<0.05), while there was no significant difference between two sides for the width of condylar cartilage. The experimental rats developed an asymmetric mandible, significantly shorter in horizontal dimension but longer in the vertical dimension on the ipsilateral side(P<0.05). The growth direction of mandible was also affected in experimental group with a more forward and upward growth on ipsilateral side. Conclusion The different shape of condylar cartilage and growth direction of mandible induced by mandibular lateral displacement on bilateral sides can be ascribed to the asymmetry of mandibular bone on two sides.

Abstract:Objective To study  the effect of vascular endothelial growth factor (VEGF) on the viability of skin flap treated with radiotherapy at different time after operation and clarify its enhancing effect on angiogenesis.Methods Fifty Wistar rat were randomly divided into 3,5,7,9 d groups and control group.A 3 cm×4 cm full thickness dorsal flap with the pedicle remaining attached at the posterior end in rats and were treated with 60Co radiotherapy at the 2nd,4th,and 6th day postoperatively.The total quantity was 12 Gy.In the experiment groups every rat was given VEGF 120 ng on the 3rd,5th,7th and 9th day.The rats in control group were given the same quality normal saline and radiotherapy.Pathological observation,fluorescence staining and  sesuccinic dehyrogenase(SDH) assay were used to measure the blood vessel density,blood vessel diameter, Microcirculation and cell vitality of skin flap.Results In 5 d and  7 d VEGF group,the number of  average blood vessels and cells of the skin falp was increased  compared with control group（P<0.05）.In  9 d group,there was a significant increase of fiber cell layer and the thickness of the skin was increased.Fluorescence staining and SDH assay showed the flap active was highest in  the 7th day group.Conclusion The density and diameter of blood vessel are higher in the 7th day group than other groups.So the time of giving VEGF should be postponed properly.

Abstract:Objective To construct the eukaryotic expression vector of human fibrinolysin for further  study on anti-thrombus and thrombolysis. Methods  Human fibrinolysin gene was amplified  from human fetal liver tissue with PCR，and was cloned into  eukaryotic expression plasmid pCI-neo and then sequenced.After the correct identification by sequencing,the eukaryotic expression recombinant plasmid of human fibrinolysin was constructed and identified with PCR and enzyme digestion.Results The PCR amplification product of fibrinolysin gene was 1 740 bp.The result of sequencing was the same as that registered in GenBank.The results of PCR and enzyme digestion showed that the recombinant eukaryotic expression plasmid had  correct codogenic gene fragment.Conclusion The eukaryotic expression recombinant plasmid of human fibrinolysin gene is successfully constructed and identified.

Abstract:Objective To investigate the association between MAOA gene polymorphism and internalizing disorders and provide theoretical basis for its susceptible gene study.Methods The method of PCR-based ligase detection reaction (PCR-LDR) was applied to detect the genotypes of the tag SNP on MAOA gene in 194 cases affected by internalizing disorders and 177 controls of healthy undergraduates.Hardy-Weinberg equilibrium for genotypic distribution was estimated by the goodness of fit χ2 test.The distribution of allelic and genotypic frequencies of MAOA gene among case groups,control group and affected different number of internalizing disorders groups was statistically computed.Results The genotypic frequency of MAOA gene did not deviate from Hardy-Weinberg equilibrium in both case and control groups (χ2 =3.763,P>0.05；χ2 = 3.213,P>0.05).The tag SNP was not associated with the disorders (P>0.05).There were no correlations between allelic or genotypic frequencies among the different degrees of co-morbidities and control group (P>0.05). Conclusion MAOA gene may not be associated with internalizing disorders,which means MAOA gene may not be the major gene caused the disease.

Abstract:Objective To explore the association between polymorphism of ACE gene and primary hypertension in Korean and Han nationalities.Methods The ACE gene polymorphism in 106 patients with hypertension(case group) and 105 healthy controls(control group) were examined by polymerase chain reaction-restriction frag
ment length polymorphism (PCR-RFLP).Hardy-Weinberg equilibrium for genotypic distribution was tested using Chi-square test of goodness of fit.SPSS13.0 was applied to analyze the association between allelic gene and genotypic distribution and primary hypertension. Results The genotypic frequency distribution of ACE gene was not deviated from Hardy-Weinberg equilibrium in both case group and control group(P>0.05).The genotypic（I,DD and ID）and allelic frequencies(I,D) of ACE gene had no significant difference between case and control groups(P>0.05).There were no remarkable differences of genotypic frequency and allelic frequency of ACE gene between case and control groups of Korean nationalities (P>0.05).There were  no remarkable differences of genotypic frequency and allelic frequency of ACE gene between case and control groups of Han nationalities(P>0.05).And the genotypic frequency and allelic frequency of ACE gene had no statistically significant difference  among Korean and Han nationalities in healthy samples (P>0.05),the genotypic frequency and allelic frequency of ACE gene  had no statistically significant difference among Korean and Han nationalities in healthy controls(P>0.05).Conclusion The polymorphism of ACE gene may not be associated with primary hypertension in Korean and Han nationalities.

Abstract:Objective To investigate the relationship between 5-HT gene polymorphism and violent behavior.Methods The techniques of PCR and restriction fragment length polymorphism (RFLP) method were used to detect the genotypes of rs6296 site of 5-HT1B gene and rs6314 site of 5-HT2A gene in  92 prison staff (case group) and 101 normal controls (control group) .The association between the polymorphisms of 5-HT gene and violent behavior was analyzed with SPSS 13.0.Results The genotypic frequencies of the 2 SNPs didn’〖KG-*3〗t deviate from Hardy-Weinberg equilibrium  both in case and control groups(P>0.05).The allelic and genotypic frequencies of the 2 SNPs had no remarkable difference between case and control groups  (P>0.05).Conclusion The polymorphism of rs6296 site of  5-HT1B gene  and rs6314 site of 5-HT2A gene  may
 be not associated with violent behavior.

Abstract:Objective To investigate the expressions of CK17 and P63 in cervical cancer tissues,and explore the possibility of CK17 and P63 as markers of cervical cancer stem cells.Methods The expressions of CK17 and P63 in cervical squamous cancer tissues (n=55)　and normal cervix tissues (n=20) were detected with immunohistochemical method.Results ①CK17 expressed in reserve cell cytoplasm in normal cervical specimens.In the cervical cancer specimens,the expression of CK17 was increased and it seemed that CK17 was stained in the margin of the cancer nest and the tumor embolus.In specimens with high grade malignant cancer or after the chemotherapy,the expressions of CK17 were increased（P<0.05）.②P63 was stained in nuclei of reserve cells and almost all cancer cells.Conclusion  CK17 is stained in the margin of the cancer nest and the tumor embolus.In specimens with high grade malignant cancer or after the chemotherapy,the expression of CK17 is increased.CK17 may be the marker of cervical cancer stem cells，but P63 not.

Abstract:Objective To investigate the genetic association between the PTEN single polymorphisms and laryngocarcinoma in Chinese Han population.Methods The method of PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis was used to detect the genotypes of rs1903858 and rs701848 of PTEN gene in 253 subjects,including 149 sporic laryngocarcinoma patients(case group) and 104 controls(control group).The association between anti-oncogene PTEN single nucleotide polymorphism and laryngocarcinoma was analyzed with SPSS10.0. Results The genotypic frequencies of the 2 SNPs of anti-oncogene PTEN did not deviate from Hardy-Weinberg equilibrium in both case and control groups.The allelic and genotypic frequencies of the 2 SNPs had no remarkable differences between case and control groups (P>0.05).Conclusion Neither rs1903858 nor rs701848 of the PTEN gene has no association with laryngocarcinoma  in Chinese Han population.

Abstract:Objective To investigate the changes of molecule markers of pre-thrombotic state,expression of CD62P,concentration of plasma fibrinogen(Fib),prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT) in the patients with essential hypertension for providing theoretical evidence for diagnosis and therapy of thrombus complication in early clinical stage. Methods Fifty-three patients with hypertension（grade Ⅰ 17 cases,grade Ⅱ 16 cases,grade Ⅲ 20 cases）were selected and twenty-eight people with medical examination in good health were used as control group.The expressions of CD62P and CD61 were measured by flow cytometry.The concentration of Fib and the levels of  PT,APTT,TT were measured by Class coagulation method.Results The expression of CD62P and the concentration of plasma Fib in essential hypertension group were higher than those in control groups (P<0.01).The levels of PT,APTT,TT in essential hypertension group were lower than those in control groups (P<0.05).The expressions of CD62P in essential hypertension grade Ⅱand Ⅲ groups were obviously higher than that in essential hypertension grade Ⅰ　group (P<0.01).The concentration of plasma Fib in essential hypertension grade Ⅲ groups was significantly higher than those in essential hypertension grade Ⅰ and Ⅱgroups (P<0.01).The levels of PT,APTT,TT in essential hypertension grade Ⅱ and Ⅲ groups were obviously lower than those in essential hypertension grade Ⅰ group (P<0.05).There was no difference of the expression of CD61 between any groups (P>0.05).Conclusion The essential hypertensive patients are in pre-thrombotic state,the activation of platelet and blood clotting activity increase.Molecular markers should be measured in essential hypertensive patients for prevention and treatment of thrombotic disease.

Abstract:Objective To observe the therapeutic effect of reformed  subhibernation therapy on  status epilepticus in patients with severe viral encephalitis. Methods 96 children with severe viral encephalitis complicated by status epilepticus were randomly divied into control group and experimental group. Control group: routine treatment was given that includes anticonvulsivus, ice compress and conventional therapy. The anticonvulsivus was applied in five drugs: the same doses of  wintermin and phenergan mixed up, 10% chlorpromazine hydrochloride，luminal, valii, the drugs were delivered when convulsion occurred otherwise not. Experimental group: in addition to routine treatment  reformed subhibernation therapy was given.The anticonvulsivus drug’s was delivered in return each 4-6 h according to the drugs half life. And anticonvulsivus drugs were applied for 2 d then gotten off.The  clinical manifestation, auxiliary examination and therapeutic efficacy of patients in two groups were observed.  Results All of 96 cases were analyzed. The lotal effective rate in experimental group is higher than that  in control group (χ2=5.872，P<0.05). All the patients had no  respiratory suppression. There was no obviously side effect in  two groups. The recovery time of epilepticus,fever, headache and vomition in experimental group was significanly shorter than those in  control group(P<0.05). The symptoms of status epilepticus in experimental  group was obviously alleviated.The recovery time of paralysis of limb,coma and aphasia of patients in experimental group was shorter than that in control group (P<0.05).
Conclusion The therapeutic effect of reformed  subhibernation therapy on  staturs epilepticus in patients with severe viral encephalitis is clear and safe.

Abstract:Objective To establish an equation between canine and other permanent teeth,and probe a new method to predict the sum of the mesiodistal diameters of unerupted permanent canines.Methods Samples consisted of 118 dental casts　were obtained from Chinese patients (50 males,68 females,respectively).Mesiodistal tooth diameters were measured by a vernier caliper.GAS was used to establish the equation between canine and other permanent teeth，and compared with stepwise regression analysis.Those were modelling samples that optimized equations coefficient(112 samples).Others were inspection samples(6 samples).Results The equation between canine and other permanent teeth was established.    MU=0.0511U1+0.2164U2+0.2780U4+0.0407U5+0.2853U6+0.8321①；ML=0.2998U1+0.1294U2+0.3912U4+0.0088U5+0.0791U6+0.9839②；FU=0.1419U1+0.1741U2+0.3258U4+0.0412U5+0.093U6+1.7355③； FL=0.2796U1+0.3750U2+0.2968U4+0.0268U5+0.0043U6+0.6030④  .Compared with stepwise regression analysis,the mean error of precision  by  GAS was less（P<0.05）.Comparing the practical predictional diameters between two methods,the absolute mean error of male was less by GAS.Conclusion The equation by GAS,which could predict diameters of unerupted canines,shows more accuracy than stepwise regression analysis.It is easy to use and valuable for orthodontic design proposal.

Abstract:Objective
To study the curative effect of ZeGuiLongShuang capsules combined with naftopidil on benign prostatic hyperplasia(BPH). Methods 140 cases with BPH were randomly divided into treatment group and control group.The patients in treatment group were given ZeGuiLongShuang capsules(once 2 tablets,3 times a day) combined with naftopidil(once 25 mg,once a day before sleeping).The patients in  control group were given only naftopidil(once 25 mg,once a day before sleeping).Internatianal-prognosic-scoring-system(IPSS),uroflowmetry-max(Qmax),Quality-of-life(QOL),residual urine and weight of prostate before and after treatment were detected.Results IPSS,Qmax,QOL and residual urine changed in two  groups.IPSS,Qmax,QOL and residual urine in treatment group were more significant than those in control group(P<0.05）.There was little change of weight of prostate.Conclusion There is no remarkable side effect of ZeGuiLongShuang capsules combined with naftopidil.ZeGuiLongShuang capsules combined with naftopidil is highly effective for BPH by decreasing rational symptom and objective index and improving the life quality of patients with BPH.

To investigate the clinical adjunctive therapeutical effect of creatine phosphate on the patients with acute severe head injury during operation to provide key material for study on its clinic therapeutic application.Methods 40 patients with acute severe head injury underwent operation were randomly divided into creatine phosphate group and control group.The patients in creatine phosphate group were injected with creatine phosphate（2 g）during operation.The patients in control group were administrated with saline i.v.with the same volume.mean arterial pressure(MAP),heart rate(HR),cardiac output(CO),stroke volume index(SI),pulmonary vascular resistance(PVR),intracranial pressure(ICP) and  brain partial pressure of oxygen(PbrO2)were recorded at pretreatment(T1),after administration (T2),after dura incision (T3),2 h after administration (T4) and after operation（T5）.The content of protein S100β　of per-patient was measured before and 24 h after operation,the cases of complication such as nausea,shivering and so on during operation were recorded after operation,according to Glasgow Outcome Scale(GOS),the prognosis of the patients was evaluated.   Results There was no markedly variation about MAP and HR between two groups on each time during opertion（P>0.05）.Compared with control group,CO and SI were significantly higher in creatine phosphate group during operation（t=4.019，P<0.05）at T2-T5,at the same time PVR in creatine phosphate group was notably lower than that in control group（t=3.517，P<0.05）. After operation,ICP in creatine phosphate group was notably lower than that in control group,at the same time PbrO2 in control group was significantly higher than that in creatine phosphate group（P<0.05）.After operation,the content of protein S100β in control group was significantly increased compared with  before operation（P<0.05）.The patients with  GOS from grade Ⅰ to Ⅴ in creatine phosphate group were 9 cases,4 cases,2 cases,3 cases and 2 cases and those were  2 cases,4 cases,6 cases,2 cases and 6 cases in control group（U=116.10，P<0.05）.The effective rate in creatine phosphate group was higher than that in control group(P<0.05).
Conclusion Administration of creatine phosphate can increase myocardium contractility in patients with acute severe head injury during operation in earlier period,and can improve the microcirculation and energy metabolism of the heart and brain constitutions,and promote the rehabilitation of patients.

Abstract:Objective To evaluate the influence of  black dots,isolated line above the best correct visual acuity and multiple lines containing the best correct visual acuity in the accuracy of Jackson cross cylinder examination.Methods Forty students were chosen (11 males,29 females,between 24 to 29 years old).Routine examination of phoropter was used to check under the maximum plus to maximum visual acuity (MPMVA) status.The sequence of three fixation targets was chosen randomly.Astigmatism chart was used to judge the best one when the different results were obtained.Results Among the 80 eyes of 40 students,12 eyes were spherical (the cylinder degree below 0.25 DC).The response of three fixation targets was the same in 19 eyes（27.9%）,23 eyes （33.8%）were sensitive to the black dots target,14 eyes （20.6%）were sensitive to the isolated line above the best correct visual acuity,12 eyes（17.6%） were sensitive to multiple lines containing the best correct visual acuity.Kruska Wallis Test was used to detect the difference among three groups.No significant difference was found (χ2 =4.026，P=0.134 ).Conclusion According to the efficiency of examination,communication and judgment,black dots target is recommended although no difference is detected among three fixation targets.

Abstract:Objective To approach the efficacy and safety of  arotinolol in treating essential hypertension. Methods The studies about arotinolol in treating essential hypertension were  accessed by searching the Cochrane Central Register of Controlled Trials(Issue 3，2008)，MEDLINE(1991 to March 2009)，EMbase(1991 to March 2009)，CBMdisc(1991 to March 2009)，and CNKI(1994 to March 2009)．The relevant journals and conference proceedings also hand searched．Randomized controlled trials (RCTs) in which arotinolol was used to treat patients with essential hypertension were collected．Then   the retrieved studies according to predefined inclusion and exclusion criteria were screened，the quality of included studies was evaluated，and Meta analysis was performed by using RevMan 4.2 software． Results A total of 176 articles were found and 6 of which were finally included.In homogeneity test：χ2=4.41，df=7，P=0.73(efficacy)；χ2=2.96，df=4，P=0.56 (safety).In combined test，Z=0.64（P=0.52），OR=1.17，OR95％CI (0.72-1.85) (efficacy)；Z=1.75（P=0.08），OR=0.60，OR95％CI (0.34-1.06) (safety).Conclusion There is no significant difference in efficacy and safety between arotinolol and control group in treating essential hypertension.

Abstract:Objective
To investigate and analyze the  prevalence rate and related risk factors of metabolic syndrome (MS) in order to explore the relationship between MS and dietary intake  in  Shuixigou Kazak people in Urumqi County Town of Xinjiang. Methods Using epidemiological investigation methods combined with laboratory test, the  dietary intake and body mass index, waist circumference, blood pressure and fasting blood glucose (FBG), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) and other clinical indicators related to MS in 131 persons were detected and the  related morbidity and risk factors were analyzed. Results The  Karaka’s dietary structure was imbalance (low fruits and vegetables intake, and high salt intake). The  prevalence rate of MS was 31.7%, of which 48.0% for men and 24.6% for women, there was a significant difference between men and women (P<0.05). The prevalence rate of MS-related clinical indicators, such as WC, HBP and L-HDL-C were highest (56.92%, 53.66% and 69.51%). Conclusion  Urumqi residents of the Karaka regions exist irrational dietary pattern; the  prevalence rate of MS is relatively high and increased with aging and has a serious threat to life and health. The reason may be low intake of fruits and vegetables, high intake of salt, meat and　animal fat intake, and the people have smoking and drinking habits. MS prevention should be focused on the people under 40-year-old and early MS can be found through a physical examination,which  has a great significance in  prevention and treatment of MS and reduction of  the occurrence.