To discuss the protective effect of zinc on spinal cord ischemia-reperfusion injury in rats and provide the experimental basis for prevention of the disease. Methods 40 healthy rats were randomly divided into four groups: high dose zinc treatment group (20  mg·kg-1),low dose zinc treatment group （10  mg·kg^-1),spinal cord ischemia-reperfusion injury  model group and sham operation group. Before the establishment of the model of spinal cord ischemia-reperfusion injury,the rats in zinc treatment groups were administered with ZnSO4 solution daily by intraperitoneal injection for 5 d,while the equal volume normal saline was given to the rats in model group and sham operation group. Then the rats were subjected to spinal cord ischemia-reperfusion injury by abdominal aorta occlusion. After 45 min ischemia and 24 h reperfusion,the neurological function evaluation (Tarlov Scale) at 24 h after reperfusion was performed. And the pathohistological changes were observed by the methods of HE staining and Fast blue staining. Results The rats in model group showed different degrees of paralysis of posterior limbs. The neurological function results showed that the  neurological function of posterior limb of rats in zinc treatment groups was improved obviously and the neurological function evaluation was significantly higher than that in  model group(P<0.01). The pathohistological changes such as cellular edema,pyknosis,paramorphia,vascular engorgement,loose and loss of myelin sheath in white matter and so on could be observed in rats in model group. But the pathohistological changes in zinc treatment groups were improved,the cell morphology was normal and the rats in high dose zinc treatment group were better than those in low dose zinc treatment group. Conclusion Zinc in certain dose range can protect spinal cord from ischemia-reperfusion injury.

Abstract:Objective To construct a recombinant plasmid pshuttle-Egr1-shTRAIL-shES containing tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) and endostatin double genes.Methods The secretary endostatin gene (shES) fragment was amplified from the pMD19T-endostatin vector by PCR. The shES gene was ligated to pMD19T and sequenced. Finally,using the gene recombinant technique,the recombinant plasmid pshuttle-Egr1-shTRAIL-shES with radiation-inducible Egr1 promoter,secretary TRAIL and endostatin double-gene was constructed.  Results The sequence of the shES gene was in concordance with that anticipated indicating shES gene was acquired successfully. Moreover,the results acquired by PCR and restrictive digestion identification of the recombinant plasmid pshuttle-Egr1-shTRAIL-shES and all the vectors refered to its construction confirmed that pshuttle-Egr1-shTRAIL-shES was constructed correctly. Conclusion The radiation-inducible double-gene co-expression vector pshuttle-Egr1-shTRAIL-shES is constructed successfully,which would set the experimental foundation for further study on the anti-tumor effect of TRAIL and endostatin double-gene-radiotherapy and its related mechanisms.

o observe the changes in the secretion of IL-12,IL-18 and the expressions of TLR4 in J774A.1 cells after ionizing irradiation,and explore the mechanism of Toll signaling pathway.Methods J774A.1 cells were divided into low dose (0.075 Gy)and high dose (2.00 Gy) irradiation groups,each group included sham irradiation,4,8,16,24,48 h  after irradiation subgroups. Flow cytometry and ELISA were used to detect the expression of TLR4 and the protein levels of IL-12 and IL-18, respectively. Results
At early stage,both of IL-12（4 h） and IL-18（4 h） expressed more than sha
m irradiation after low dose ionizing radiation,and then gradually returned to the   level in sham irradiation group. After high dose ionizing radiation,the expression of IL-12 kept higher level compared with  sham irradiation group at 4 -24 h（P<0.05 or P<0.01）,and then returned to the  normal level at 48 h;while the expression of IL-18 was similar with that in low dose  irradiation group,it was up-regulated at 4 h after irradiation（P<0.05）,and then gradually returned to the control level. Conclusion Ionizing radiation  can　increase the expression of TLR4 and the protein levels of IL-12,IL-18 and promote the immune response and start innate immunity.

Abstract:Objective To investigate the effects of berberine on vascular endothelial function in type 2 diabetic rats and explore the prophylactic and therapeutic significance and pharmacological mechanism of berberine in type 2 diabetic vascular complications. Methods Wistar rats were randomly divided into four groups: diabetic group,control group,diabetic rats treated with berberine (100 mg·kg-1) group,and control rats treated with berberine group. The serum fasting blood glucose(FBG),insulin,total cholesterol(TC),triglyceride(TG) and fasting insulin(FINS) levels were tested. Acetylcholine (ACh)-induced endothelium-dependent relaxation and sodium nitroprusside
-induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. In addition,the histopathology measurement was  performed with  hematoxylin and eosin staining. Results Compared with diabetic group,the FBG,TC and TG levels in diabetic rats treated with berberine were significantly decreased,meanwhile the insulin sensitivity index(ISI) was increased. The  Ach-induced endothelium-dependent relaxation in the aorta from diabetic rats was  significantly improved in berberine treatment group,while sodium nitroprusside (SNP)-induced endothelium-independent relaxation showed no difference among four groups. The histopathological results showed that berberine attenuated the  damage of the aortic endothelium in diabetic rats. However there was no difference in these indexes between control group and berberine treated control group. Conclusion Berberine restores diabetic endothelial dysfunction through decreasing the FBS and blood lipid,improving insulin resistant and enhancing NO bioavailability.

To investigate the influence of silenced poly-（ADP-ribose）glycohydrolase（PARG）on the expression and phosphorylation of protein kinase P38 in colorectal carcinoma lovo cells and clarify its mechanism.  Methods Lentivirus PARG-shRNA was transfected into colorectal carcinoma lovo cells，then the lovo cells with silenced PARG was perpetually selected. Untreated lovo cells were used   as positive control(untransfected group)，the lovo cells  treated with empty vector were used as   negative control  (empty-vector-transfected group)，the lovo cells   treated with PARG-shRNA were used  as   experimental comparison(PARG- shRNA transfected group). The expression of PARG mRNA was detected by RT-PCR. The expressions of PARG，poly-（ADP-ribose）polymerases（PARP），P38 and p-P38 protein were detected by Western blotting.  Results Both RT-PCR and Western blotting results showed that the expressions of PARG in PARG-shRNA transfected group was obviously lower than that in untransfected group（P<0.05）. The expressions of PARP，P38 and p-P38 in PARG-shRNA transfected group were obviously lower than that in untransfected group（P<0.05）．
Conclusion The silenced PARG can inhibit the expression and phosphorylation of P38 in human colorectal carcinoma lovo cells. It  may be related to down-regulating PARP .

To investigate the effect of poly (ADP-ribose) glycohydrolase (PARG) inhibitor gallotannin on renal lesion in STZ-induced diabetic rats and its mechanism. Methods Wistar rats were randomly divided into five groups: normal control (NC) group,STZ-induced diabetic(DM)  group,diabetic rats treated with 3-aminobenzamide (CDT) group,diabetic rats treated with gallotannin 20 and 30  mg·kg^-1·d^-1( LDT and HDT) groups. The levels of blood glucose,serum creatinine,blood urea nitrogen,urinary protein excretion were detected after treatment for 12 weeks.The  renal morphology was observed and the expression of poly (ADP-ribose) polymerase (PARP) was tested by immunohistochemical method. Results Compared with DM group,the levels of blood glucose，serum creatinine,blood urea nitrogen  and urinary protein excretion were significantly decreased in both LDT and HDT groups(P<0.05 or P<0.01). The renal lesion and the signal of positive expressing PARP in renal tissues were markedly reduced in diabetic rats treated with GLTN.But there were no significant differences of the indexes mentioned above between LDT and HDT groups(P>0.05).Conclusion Gallotannin can lower the level of blood glucose,inhibit the  renal PARP expression,and ameliorate the renal functional and morphological abnormalities in diabetic rats.

To study the protective effect of paraxadiol saponins(PDS) on myocardium of rats with endotoxic shock and reveal the relationship between the protective effect and AQP1 expression level. Methods 40 Wistar rats were divided into 4 groups: control group(CTR),lipopolysaccharide endotoxic shock model group(LPS),dexamethasone treatment group(DEX) and PDS group. LPS (5 mg·kg-1) was administered sublingually to build up the model of endotoxic shock,the mean arterial pressure(MAP) and the decreasing percentage of MAP were monitored at different time. The contents of AST,CK and LDH in serum and SOD activity and MDA content in myocardium were detected after shock for 240 min.The AQP1 expression level in myocardium was detected with Western blotting. Results When the blood pressure fell to 2/3 of the baseline it was regarded as the start of shock. It was found by dynamic observation that the rats entered into shock state at 5 min,the MAP had an compensatory restoration at 20 min,the MAP fell progressively since 60 min until the models got close to death at 240 min in LPS group,but  the MAP remained steady and constant compensatory changes in PDS and DEX groups until the rats were sacrificed at 240 min. The serum contents of AST,CK and LDH in LPS group were highest among groups,but in PDS and DEX groups they were lower (P<0.05)；the SOD activities in PDS and DEX groups were higher than those in LPS group (P<0.05),meanwhile the contents of MDA were lower than those in LPS group(P<0.05). Western blotting result showed that the quantity of expression in LPS group was significantly lower than that in control group(P<0.05),but in PDS group and DEX group they were higher than that in LPS group(P<0.05). Conclusion PDS has the protective effect against myocardium injury in rats with endotoxic shock,the mechanism may be adopted to enhance the expression of AQP1 in capillary endothelial cells of LPS-induced endotoxic shock rats,and inhibit the myocardial stromal edema.

Abstract:Objective To investigate the influence of gross saponin tribulus terrestris (GSTT) on nuclear factor-κB (NF-κB) of pheochromocytoma cells (PC12 cells) injured by hypoxia and its mechanisms. Methods PC12 cells were divided into control group, model group, high dose GSTT group（10 mg·L^-1） and low dose GSTT group（3 mg·L^-1）. The hypoxia model was set up with H2O2. The apoptotic rate of PC12 cells was detected by flow cytometry. The protein expression of NF-κB was detected by flow cytometry, immunohistochemistry and Western blotting methods. Results Compared with model group, the apoptotic rates of PC12 cells in high dose GSTT group and low dose GSTT group were significantly reduced (P<0.05). Compared with low dose GSTT group, the apoptotic rate of cells in high dose GSTT  group was reduced (P<0.05). The results of flow cytometry and Western blotting showed that the NF-κB expressions in high dose GSTT group and low dose GSTT group were significantly increased compared with model group (P<0.05). There was no significant difference of the NF-κB expression between high dose GSTT group and l
ow dose GSTT group. The results of immunohistochemistry showed that few brown and black grains were found in the cytochylema in model group and the NF-κB expression was reduced compared with control group(P<0.01). Compared with model group, the NF-κB expressions in high dose GSTT group and low dose group were significantly increased (P<0.05). Conclusion GSTT can protect PC12 cells injured by hypoxia which might be correlated with the activation of nuclear factor-κB expression.

Abstract:Objective To observe the influence of  aprotinin in   ultrastructural changes of liver tissues in mice with experimental acute liver injury and   explore its  protective effect on  liver tissues.  Methods The mouse medels with carbon tetrachloride (CCl₄) liver injury were established and the mice were divided into normal control group,model group,ganlixin injection groups and low,  middle  and high dose aprotinin groups. The tissue specimens were obtained from various groups,uranium oxide-acetate-citric acid-lead double staining was used to observe the changes of liver tissues under TEM.  Results In model group,  the liver cells were  swelled with pyknotic nucleus,reduced intracytoplasmic rough endoplasmic reticulum,pyknotic mitochondria,and cavitation. In  low  dose aprotinin group,the  individual cells were  pyknotic with a few rough endoplasmic reticulum and lysosomes in liver cells,timid Endoluminal microvilli disappeared.In middle  dose  aprotinin group, the size of  liver cells was a little larger,the liver cell nucleolar was evident with intracytoplasmic rough endoplasmic reticulum, mitochondria,numerous glycogen accumulation and more timid Endoluminal microvilli. In high dose aprotinin group,the liver cell structures were similar to those in control . The ultrasturctural changes in ganlixin injection group were similar to those in middle dose aprotinin group.  Conclusion The intraperitoneal injection of aprotinin has significantly protective effect on the ultrastructures of liver tissues in acute liver injury mouse models.With the increase of aprotinin,the ultrastructural changes of  liver tissues are markedly reduced.

To investigate the effect of cAMP on fatty acid oxidation in skeletal muscle cells and elucidate the mechanism of the effect of cyclic AMP(cAMP) on fatty acid oxidation in skeletal muscle. Methods The primary muscle cells(PMC) and C2C12 cells were treated with α-MSH,cAMP analogs Bt2cAMP,8-BrcAMP and cAMP antagonist respectively,and they were divided into α-MSH group,cAMP agonists group,cAMP antagonist plus α-MSH group,and control group was set up at the same time.The fatty acid oxidation was measured by quantitating the production of 3H2O from (9,10 -3H) palmitate. Melanocortin receptor(MCR) subtypes in the muscle and C2C12 cells were measured by RT-PCR. Results The fatty acid oxidation was  significantly increased in  cAMP analogs group and α-MSH group   compared with  control group(P<0.05),and the fatty acid oxidation in  cAMP antagonist plus α-MSH group was significantly reduced  compared with  α-MSH group(P<0.05),and there was no significant difference compared with  control group(P>0.05). Conclusion α-MSH and cAMP analogs significantly increase the fatty acid oxidation,α-MSH might be binding with MCR and they increase fatty acid oxidation in skeletal muscle through cAMP signal transduction pathway.

To study the effect of ginsenoside Rb3 (G-Rb₃) on experimental ventricular remodeling  in rats and investigate its mechanism.  Methods The ventricular remodeling model was induced by myocardial infarction in rats of left anterior descending coronary occulusion. The Wistar rats were divided into 4 groups:sham operation group,remodeling model group,G-Rb3 group,and captopril group. The rats in sham operation group and remodeling model group were treated with normal sodium (with 2mL·kg^-1·d^-1 i.p).The rats in G-Rb₃ group were treated with G-Rb₃ (with 20 mg·kg^-1·d^-1 i.p). The rats in captopril group was treated with captopril (100 mg·kg^-1·d^-1 i.g). After 4 weeks,the morphological parameters,histopathology,hemodynamic changes,biochemical analysis were determined in rats. The content of malondialdehyde (MDA) in serum and activities of superoxide dismutase(SOD),glutathione  peroxidase (GSH-Px) in plasma,the content of endothelin (ET),angiotensin Ⅱ(AngⅡ) in plasma,myocardial angiotensin Ⅱ(AngⅡ) were determined.  Results Compared with model group,the cardiac coefficient,the left ventricular end diastolic pressure (LVEDP) were decreased significantly(P<0.01),the mean arterial blood pressure(MAP) and the maximum left ventricular presure rising and dropping rates (±dp/dtmax) were increased significantly in G-Rb₃ group(P<0.01). In addition,Compared with model group,the contents of MDA in serum,ET,AngⅡ in plasma and myocardial  AngⅡ were declined(P<0.01),then the activities of SOD and GSH-Px in serum were increased markedly in G-Rb₃group(P<0.01).  Conclusion G-Rb₃ has protective effect on ventricular remodeling through inhibitting renin-angiotensin-aldosterone system(RAAS) to reduce contractive material and  reinforcing  ability of anti-oxidant

To compare of immunological efficacy of H1N1 influenza vaccine 0 d one-dose method and 0,21 d,two-dose method,and observe whether seasonal influenza vaccine has cross protection  for H1N1 influenza virus. Methods H1N1 influenza vaccine with two immunological methods was used to inoculate Balb/c mice.The first,the Balb/c mice were inoculated in 0 d. The second,the Balb/c mice were inoculated in 0 and 21 d. The serum from orbital in 14,21,28,35, 42 d after the first-dose were separated. HI was performed to determine the H1N1-specific serum antibody titers to compare the immune effect of the two immunological methods. Then the Balb/c mice were inoculated with seasonal influenza vaccine. The serum from orbital in 14 and 28 d were separated. HI was performed to determine whether the anti-serum of seasonal influenza vaccine could non-specificly bind with H1N1 influenza virus-specific antigens. Results The 28,35  and 42 d antibody titers of the two-dose method were significantly higher than those of  one-dose methods after the second immunity at the 21th day. And the 42 d antibody titers  of  the two-dose methods were 4 times of that of  one-dose methods. HI was performed to determine the anti-serum of seasonal influenza vaccine,there were  is no H1N1 influenza virus-specific antibodis in it. Conclusion The antibody titers of 0 and 21 d two-dose method still maintain the high level at  28th day,and  are obviously higher than those of one-dose methods. There is no cross-protective immunity between seasonal influenza vaccine and influenza A H1N1 influenza virus.

To investigate the inhibitory effect of 20(S)-proto-panaxdiol（PPD）on the growth of siha cells in vitro and detect the transcription and expressions of autophagy related gene Beclin 1 and MAP1-LC3,and clarify the mechanism of its suppressing effect on siha cell proliferation. Methods The siha cells cultivated in vitro were treated with alcohol,40 μmol·L^-1 platinol (Pt),10 μg·L^-1 PPD,20 μg·L^-1 PPD,40 μg·L^-1 PPD for 24,48,72 and 96 h, respectively. The inhibitory rate of cell proliferation in each group was detected by MTT,the transcription and expressions of autophagy related gene Beclin 1 and MAP1-LC3 were analyzed by RT-PCR,Western blotting and immunocytochemical staining. Results After treated with 10,20 and 40 μg·L^-1 PPD,the inhibitory rates of siha cells were significantly increased compared with control group(P<0.05),and the inhibitory rate was  increased gradually with the time prolongation of time and the increase of dose. The transcription and expressions of autophagy related gene Beclin 1 and MAP1-LC3 were significantly up-regulated (P<0.01). Conclusion PPD may suppress the proliferation of siha cells,and its mechanism may be related to  up-regulating the expressions of autophagy related gene Beclin 1 and MAP1-LC3.

To investigate the induction  of fluvastatin (Flu) on apoptosis of human promyelocytic leukemia cells (HL-60 cells) and its mechanism,and  provide theoretical basis for clinical application of Flu. Methods The cultured HL-60 cells in vitro were divided into blank control,positive control group (ATRA,10  μmol·L^-1),Flu (20  μmol·L^-1) group and Flu (20  μmol·L^-1)+ mevalonate (Mev,100  μmol·L^-1) group. After HL-60 cells were treated,the caspase-3 activity was determined using CaspACETM Assay System for exploring the  apoptosis,the apoptosis was analyzed by flow cytometry after annexin V-FITC/PI double staining,the laddered pattern of DNA fragments was evaluated by DNA agarose gel electrophoresis and the ultrastructural changes of HL-60 cells were observed by transmission electron microscope. Results After Flu treatment for 24 h,the caspase-3 activity in HL-60 cells was significantly higher (A405,30.68 ±1.57) than that in control cells (12.05±2.15,P<0.01). In HL-60 cells co-treated with Flu and Mev,the caspase-3 activity was markedly reduced (14.62 ±1.36),nearing to the level in control cells (P>0.05). Flow cytometry showed that compared with control,the apoptotic rate in Flu-treated HL-60 cells was significantly increased (4.24%  vs  10.76%,P<0.01),and the apoptotic rate was decreased to 5.11% after HL-60 cells were co-treated with Flu and Mev (P>0.05). DNA agarose gel electrophoresis revealed a laddered pattern of DNA fragments in Flu-treated HL-60 cells after 72 h treatment. In Flu-treated HL-60 cells,electron microscopic study exhibited the diminished cell body,condensed cytoplasm and nuclear chromatin condensation aggregating under the nuclear membrane,and the intact cell membrane and organelle. Conclusion Flu can induce the apoptosis of HL-60 cells and this effect is dependent on mevalonate(Mev) pathway,suggesting that the inhibition
of Mev pathway may be one of molecular mechanisms for Flu-induced apoptosis of  HL-60 cells.

To investigate the expression of p53  in myocardium tissue of ispproterenol (Iso)[KG-*3]-induced myocardial fibrosis  rats and its signigicance and clarify the effect in inyocardial of fibrosis. Methods 25 male Wistar rats were randomly divided  into five groups: control group，model groups(12 h,24 h,1 week and 3 weeks)(n=5). Except control group,all the rats in the other groups were treated with Iso to bulit rat myocardial injury models(15 mg·kg^-1).The expressions of P53 protein and p53 mRNA were detected with immunohistochemical staining and RT-PCR.Results The immunohistochemical results showed that the P53 protein expressions were  increased 12 and 24 h after Iso injection  compared with  control group（P<0.05）and the  protein expressions were obviously increased  1 week and 3 weeks after Iso injection（P<0.01）.The RT-PCR results showed that the p53 mRNA was obviously increased  at 12 h (P<0.05),arrived a peak at 24 h (P<0.01)　and was gradually decreased at 1 week,but still higher than normal level (P<0.05). Conclusion The P53 protein   expression is increased in rat myocardium tissue and the expression of p53 gene is up-regulated,suggesting that p53 has an important role in rat myocardial fibrosis.

To study the influence of quetiapine and venlafaxine in neuronal injury after cerebral ischemia and stress and illuminate the protective effects of quetiapine and venlafaxine after global cerebral ischemia and stress．Methods  Male ICR mice were pretreated with venlafaxine or quetiapine for 14 d.On the 15th day,the bilateral common carotid arteries of mice were occluded.This experiment was divided into sham group(Saline),sham plus stress group(Saline+Sham),ischemia plus stress group (Saline+ Ischemia+Stress),quetiapine plus ischemia plus stress group (QUE+Ischemia+Stress) and venlafaxine plus ischemia plus stress group (VEN+ Ischemia+Stress).The neuronal injury after cerebral ischemia  and stress was detected with immunohistochemical and Nissle  staining．Results Compared with sham group,the NeuN positive cells  were not significantly changed in sham plus stress group(P>0.05);compared with sham group or sham plus stress group,the NeuN positive cells were significantly decreased in ischemia plus stress group（P<0.05）；compared with ischemia plus stress group,the NeuN positive cells  were not significantly changed in venlafaxine plus ischemia plus stress group (P>0.05);compared with ischemia plus stress group,the NeuN positive cells were significantly increased in  quetiapine plus ischemia plus stress group（P<0.05）. Compared with sham group,the Nissle staining positive cells  were not significantly changed in sham plus stress group(P>0.05);compared with sham group or sham plus stress group,the Nissle staining positive cells were significantly decreased in ischemia plus stress group（P<0.05）；compared with ischemia plus stress group,the Nissle staining positive cells were not significantly changed in venlafaxine plus ischemia plus stress group (P>0.05);compared with ischemia plus stress group,the Nissle staining positive cells were significantly increased in quetiapine plus ischemia plus stress group（P<0.05）.Conclusion Quetiapine can attenuate the neuronal injury and has protective effects in mouse models after global cerebral ischemia and stress.

To investigate the expression of tissue transglutaminases (tTG) in rat ischemic myocardium and the role of sodium ferulate (SF) in rat myocardial ischemia. Methods 70 male Wistar rats were  randomly divided into fourteen group(n=5),control groups，model groups(2 h,4 h,6 h,12 h,24 h,48 h,72 h,1 week and 3 weeks after injection of isoproterenol hydrochloride(Iso))( the control group and 3 weeks group were divided into two groups)，positive control group and SF group. Except control group,all the rats in the other groups were treated with Iso (15 mg·kg^-1).In addition,the rats in positive control group were injected synchronously with bean and the rats in SF group were injected synchronously with SF. The tTG expression was observed before and after SF treatment by immunohistochemistry and RT-PCR. Results The tTG protein expression in rat ischemic myocardium 2 h after Iso injection was　gradually  increased 　compared with control group(P>0.05). With the prolongation of time of Iso injection,the tTG protein expression arrived a peak at 3 week compared with control group(P<0.01). The tTG mRNA expression at 24 h reached peak compared with control group and was  gradually decreased　at 48 h（P<0.05 or P<0.01）. The tTG protein expression after SF intervention was gradually decreased 　compared with model group(P<0.01). The tTG mRNA expression was also  gradually decreased　compared with model group（P<0.05 or P<0.01）.Conclusion The increased tTG expression accelerate the process of myocardial ischemia in rat myocardial ischemic model. SF can inhibit the tTG expression and reduce the process of myocardial ischemia.

To investigate the time-course effect of glutamate on dopaminergic neurons injury in mouse mesencephalic dissociated culture, and  elucidate  the effect of the neurotoxicity of glutamate on dopamingergic neurons injury in Parkinson’〖KG-*3〗s  disease(PD) model.  Methods Mouse embroys of 14 d were used to make mecsencephalic dissociated culture. The experiment was divided into control group and glutamate group.The cultures were treated with 500  μmol·L^-1 of glutamate for 15 min on the 10th DIV,and stained by TH immunocytochemistry 2,4,6,24 and 48 h after treatment. The numbers of dopaminergic neurons and the average neurite numbers per dopaminergic neuron were evaluated by counting under microscope. Results The TH-ir neurons were counted and there were 778.0± 18.6 dopaminergic neurons per well in control group; there were 314.0±15.4,298.0±19.1,307.0±18.5,and 323.0± 23.8 dopaminergic neurons per well respectively at 2,4,6,24 and 48 h in glutamate group,the TH-ir neurons in glutamate group were significantly reduced compared with  control group(P<0.05).The average numbers of neurites per dopaminergic neuron were 3.440±0.106 in control group,2.24 0± 0.105,2.390±0.103,3.070±0.105,3.420± 0.987 and 3.420±0.923 respectively at 2,4,6,24 and 48 h in glutamate goroup. There were a significantly lower neurite numbers 2 and 4 h after glutamate treatment,as compared with all the other groups (P<0.05). Conclusion The loss of dopaminergic neurons induced by glutamate is caused by a relative acute neural death and is irreversible. But
 the number of neurite and length of neurites are gradually improved as the recovery period is prolonged.

To observe if the bone marrow mesenchymal stem cells(BMSCs) cocultured with pancreatic cells can be transformed into islet-like cells,and clarify the effect   of blood glucose control  of  BMSCs in diabetic rats. Methods BMSCs and pancreatic cells were  concultured in vitro.18 SD rats were introperitonally injected with streptozotocin(STZ) to set up animal models of diabetes and the rat models were randomly divided into PBS group,BMSCs group and BMSCs cocultured with pancreatic cells group(n=6).The bromodeoxyuridine(Brdu)-labeled cells were transplanted into STZ-inducd diabetic rats by caudal vein,the clinical efficacy was observed.The  distrubition of Brdu-labeled cells  in the pancreas was examined by immunohistochemistry. Results The blood glucose in caudal vein was detected by blood glucose detection instrument.The blood glucose level  in PBS group didn’t change compared with before transplantation(P>0.05).The blood glucose level  was reduced from the 14th day after transplantation in  BMSCs group and BMSCs cocultured with pancreatic cells group,there were significant differences compared with before transplantation (P<0.01).There was  significant difference of the blood glucose level after transplantation between BMSCs group and BMSCs cocultured with pancreatic cells group(P<0.01).The  immunohistochemical results showed that the Brdu-labeled cells with positive  insulin-expression  were tested in the BMSCs group and BMSCs cocultured with pancreatic cells group,but they were  not found in  PBS group. Conclusion The cultured cells could reduce hyperglycemia in the STZ-induced diabetic rats by caudal vein.BMSCs could also reduce hyperglycemia in the STZ-induced diabetic rats by caudal vein.The   effect of the later was better than the former.

To study the protective effect of Acanthopanax senticosus saponins （ASS）on human umbilical vein endothelial cells(HUVECs) injured by advanced glycosylation end products(AGE) and its possible mechanism. Methods AGE-modified bovine serum albumin(AGE-BSA) was prepared by incubating the BSA with a high concentration of glucose.The cultured HUVECs were divided into six groups:control group,BSA group,AGE group,low dose ASS + AGE group,middle dose ASS + AGE group,high dose ASS + AGE group.The HUVECs were cultured with 200 mg·L-1 AGE-BAS for 24 h following pre-incubation of ASS at different concentrations.The HUVECs proliferation  activity, the Von Willebrand factor(vWF), lactate dehydrogenase (LDH) and nitric oxi
de (NO) levels were determined. Results Compared with BSA group,the HUVECs proliferation  activity in AGE group was significantly reduced(P<0.01),the vWF and LDH levels  in AGE group were significantly increased(P<0.01) and the NO level were significantly reduced(P<0.01).Compared with AGE group,the HUVECs proliferation activities in ASS 10,50,100 mg·L^-1 groups were increased(P<0.05).The vWF and LDH levels were markedly decreased(P<0.05). The NO level was increased(P<0.05).Conclusion ASS could protect HUVECs against damage induced by AGE and increase the NO level in the AGE- exposed HUVECs.The mechanism may be related to increasing the NO level in HUVECs.

To induce the mutation  of fibroblast growth factor 21 (FGF21)gene at the site of the 20th amino acid,express it in the E.coli BL21(DE3),purify the expressed product of FGF21［Tyr²⁰］ protein and detect the bioactivity.Methods  Using plasmid of wide FGF21 as a template,the site-specific mutagenesis of the 20th amino acid Tyr to Phe was performed by PCR,and it was fused with small ubiquitin-like modifier(SUMO) and cloned into vector pET22b. The constructed recombinant plasmid pET22b-SUMO-FGF21［Tyr²⁰］ was transformed to E.coli BL21(DE3) for expression under induction of IPTG. The expression product was identified with SDS-PAGE and Western blotting,purified with Ni-NTA affinity column,the SUMO was cut with SUMO enzyme and demineralizated with Sephadex G50.Results The site-specific mutagenesis of the 20th amino acid of FGF21 was made by PCR successfully.The  recombinant plasmid pET22b-SUMO- FGF21［Tyr²⁰］ after identification  by PCR,digestion and sequencing analysis  matched the expected result. Through SDS-PAGE electrophoretic analysis,the protein was soluble. After purification the  FGF21［Tyr²⁰］ mutant protein was successfully obtained.Western blotting analysis showed that the FGF21［Tyr²⁰］ mutant protein had specific reaction with FGF21 antibody.Conclusion The SUMO- FGF21［Tyr²⁰］ mutant gene is successfully constructed and expressed with high solubility,and the FGF21［Tyr²⁰］ protein is also purified and obtained.

Objective To study the changes of CD4⁺CD25⁺Treg cells and immunesurveillance function in tumor infiltrating lymphocytes(TIL) and investigate the relationship between CD4⁺CD25⁺Treg cells and tumor immune escape. Methods TIL and spleen cells of the S180 ascites carcinoma mice and spleen cells of the control ones were used for the following detection: the quantities of CD4⁺CD25⁺Treg cells,CD8⁺T cells,NK cells and NKG2D were detected by flow cytometry;the expression level of Foxp3 mRNA was detected by RT-PCR;the killing function of CD8⁺T cells was detected by lactate dehydrogenase release assay. Results Compared with control mice，the quantity of CD4⁺CD25⁺Treg cells and the expression of Foxp3 mRNA were significantly increased in the TIL and spleen cells of the tumor mice (P<0.05);furthermore,the quantity of CD4⁺CD25⁺Treg cells and the expression of Foxp3 mRNA in TIL were both higher than those in spleen cells of tumor mice;the number of CD8⁺T cells in TIL was significantly increased(P<0.05),while the killing function of CD8⁺T cells in TIL was decreased;the number of NK cells and the expression of NKG2D in TIL were significantly increased (P<0.05). Conclusion The quantity and function of CD4⁺CD25⁺Treg cells are significantly increased in location of tumor which might be one of the mechanisms of tumor escaping from immune surveillance.

To investigate the regulatory role of E6-associated protein(E6-AP) in anadrogen receptor(AR) and AKT expressions in prostate cancer LNCaP cells and its mechanism. Methods  LNCaP cells stably transfected with E6-AP were used to analyze the regulatory role of E6-AP in AR and AKT expressions by Western blotting assay,the  intracellular localization of E6-AP and AR was determined by immunofluorescence. Results Compared with LNCap cells with low E6-AP expression, the total AR and phosphorylated-AR expression levels in the  LNCaP cells with high E6-AP expression were significantly decreased（P<0.05）,the  total Akt and phosphorylated-Akt expression levels were significantly increased（P<0.01）.The immunofluorescence results demonstrated that E6-AP showed a similar intracellular location with AR in the LNCaP cells.  Conclusion Overexpression of E6-AP could down-regulate the protein level of total AR and phosphorylated-AR,whereas up-regulate the protein level of total Akt and phosphorylated-Akt. E6-AP shows colocalization with AR in the LNCaP cells.

To study the neuroprotective effect of erdosteine on hippocampal neurones in rats with epilepsy and explore the relationship between oxidative stress and neuron apoptosis in rats with epilepsy. Methods 30 SD rats were randomly divided into there groups  PTZ group (n=10),erdosteine pre-disposal treatment group (n=10) and control group (n=10).The epilepsy models were established by injecting intrapertoneally pentylenetetrazol(PTZ) into the rats. The rats in   erdosteine group were pre-treated by injecting intrapertoneally erdosteine. The  behavior of rats were observed. The level of malondialdehyde(MDA),the activities of glutathion peroxidase(GSH-Px) and superoxide dismutase(SOD)were detected by colormetric method.The damage of hippocampal neurones was observed by HE and terminal-deoxynucleotidyl transferases mediated nick end labeling（TUNEL） method. Results The seizure period in erdosteine group was significantly shorter than that in PTZ group (P<0.05). The activities of SOD and GSH-Px in  erdosteine group were significantly higherer than those in PTZ group(P<0.05 or P<0.01),the MDA level in erdosteine group was significantly lower than that in PTZ group (P<0.01). Compared with PTZ group,there were less TUNEL positive cells and acidophilic neurons in the hippocampus in erdosteine group(P<0.01). In PTZ group and erdosteine group,the TUNEL positive cells had negative correlation with the activity of SOD（r=－0.482,P<0.05） and  positive correlation with the MDA level（r= 0.875,P<0.01）. Conclusion Erdosteine could reduce the oxidative stress level of rats with epilepsy,reduce seizure periods and neuronal damage. Erdosteine has the effect of brain protection probably by scavenging free radical.

Abstract:Objective To explore the expressions of STAT3 mRNA and STAT3 protein and study their correlations with the occurrence,development,invasion,and metastasis of oral squamous cell carcinoma (OSCC). Methods In situ hybridization (ISH) and immunohistochemistry were used to detect the expressions of STAT3 mRNA and STAT3 protein  in 60 cases of normal oral mucosa,30 cases of atypical hyperplasia and 60 patients with oral squamous cell carcinoma,at the same time the clinical information was combinated for analysis. Results The positive expression rates of STAT3 mRNA  in normal mucosa,atpical hyperplasia and oral squamous cell carcinoma  were 25.0% (15/60),40.0% (12/30),68.3% (41/60),respectively;there were  significant differences between various groups (χ2 = 23.855,P<0.05). The STAT3 mRNA expressions between different differentiation,different depths of invasion and lymph node metastasis in oral squamous cell carcinoma had significant differences (χ2=15.912，χ2=25.616，χ2=7.455，P<0.05）. The STAT3 protein expression rates were dramaticlly different at different grades of cancer (grade Ⅰ 40.0%,gradeⅡ 73.9%,grade Ⅲ 90.9%;χ2=11.532， P<0.05).The STAT3 protein expression rate was significantly different between shallow infiltraion cancer and deep infiltration cancer（26.3% vs 92.7%，χ2=24.989，P<0.05）. The STAT3 protein expression rate was also dramaticlly different between  lymph node metastasis cancer and non-lymph node metastasis cancer (91.3% vs 59.5%，χ2=5.602，P<0.05）.The STAT3 protein expression rates in normal mucosal tissues,atypical hyperplasia and cancer rates were  13.3% (8/60),30.0% (9/30),and 71.2% (43/60),respectively;the differences between various groups were　significant (χ2 = 23.855,P<0.05).The STAT3 protein expression was closely related with　  cancer tissue differentiation,depth of invasion and lymph node metastasis  (χ2 = 11.532,χ2 = 24.989,χ2 = 5.602,P<0.05). Conclusion The STAT3 mRNA and protein are over-expressed in oral squamous cell carcinoma tissue and have close correlations with biological behavior,occurrence and development of oral squamous cell carcinoma . STAT3 may be a new assistant indicator for early diagnosis and prognosis judgement of oral squamous cell carcinoma.

Abstract:Objective To evaluate the effect  of porcelain-fused-to-metal(PFM) crowns with different metal alloy base on the gingiva by detecting the IL-6 level in gingival crevicular fluid(GCF). Methods Ni-Cr alloy,gilt Ni-Cr alloy,gold alloy PFM were employed to total of 60 clinical patients. 6 months later,GCF from the patients was collected and quantified,and the IL-6 level was detected by ELISA kit. Results There was no significant difference of the GCF  between gold alloy group and  control group(P>0.05);the GCF in  Ni-Cr alloy group was much higher than those in gold alloy  group and  control group (P<0.05),and there was no significant difference between Ni-Cr alloy group and  gilt Ni-Cr alloy group (P>0.05). The IL-6 levels in GCF in Ni-Cr alloy group was much higher than that in  control group (P<0.05), there were no significant differences between gilt Ni-Cr alloy group,gold allog group and  control group(P>0.05).There was no significant difference  of IL-6 level between gilt Ni-Cr alloy group  and gold alloy group (P>0.05),the IL-6 levels in  gilt Ni-Cr alloy group  and gold alloy group were much lower than that in Ni-Cr alloy  group(P<0.05). Conclusion  Ni alloy has the most serious stimulation,and gold alloy has the least effect on the gingiva.The IL-6 level in GCF could be a theoretical index to monitor clinical effect of PFM.

Abstract:Objective To study the effects of blood glucose concentration  on the levels of IL-6 and platelet derived growth factor(PDGF) before and after operation and short-term prognosis in patients with acute myocardial infarction(AMI) treated by primary percutaneous coronary intervention (PCI),and discuss the possible mechanisms of those effects. Methods A total of 54 patients with AMI treated with successful primary PCI within 10 h after onset of symptoms were divided into three groups:  groupⅠ,non-diabetic patients with blood glucose level<8.0 mmol·L-1;group Ⅱ,non-diabetic patients with blood glucose level ≥8.0 mmol·L-1;group Ⅲ,diabetic patients. The method of enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of blood serum IL-6 and PDGF  before PCI 10 min and 24 and 48 h after   PCI. Results There were no  differences in the incidence rates of serious arrhythmia in operation and heart failure duration of hospital between three groups（P<0.05）,but the incidence rates in group Ⅲ and group Ⅱ were higher than that in group Ⅰ. The levels of IL-6 and PDGF were increased with blood glucose level in three groups,group Ⅲ >group Ⅱ> group I,there were significant differences (P<0.05). The level of IL-6 48 h after PCI in group Ⅰ was decreased to the level before PCI,the descent in group Ⅱ and Ⅲ was slow. The  PDGF levels  24 and 48 h after PCI had been little higher than before PCI in group I，the level of PDGF in group Ⅱ was lower than that in group Ⅲ and higher than that in group Ⅰ(P<0.05). Conclusion Stress hyperglycemia in patients with AMI treated by primary PCI maybe increase the levels of inflammatory reaction，extend the time of inflammatory reaction，increase the incidence rates of serious cardiac arrhythmia in operation and heart failure duration of hospital stay.

Abstract:Objective To detect the serum levels of cyclooxygenase-2(COX-2)in patients with  breast cancer before and after operation,and approach the relation of COX-2 with the clinicopathological characteristics of  breast cancer.Methods The  serum COX-2  expression levels  in 30 cases of breast cancer,13 cases of adenosis and 13 cases of controls  were detected by enzyme-linked immunosorbent assay (ELISA).Results The serum levels of COX-2 in pre-operative group and post-operative group were significantly higher than those in control group and adenosis group (P<0.01).The serum level of COX-2 in patients with breast cancer in pre-operative group was higher than that in post-operative group (P<0.01),the serum level of COX-2 in patients with lymph node metastasis was significantly higher than that in patients without lymph node metastasis(P<0.05),ER-negative patients had higher level of serum COX-2 than ER positive patients (P<0.01),the serum COX-2 level in HER-2 positive patients was higher than that in negative patients(P<0.01).Conclusion The serum level of COX-2 in breast ancer patients is significantly higher than those in normal control group and breast adenosis group.Detection of the serum level of COX-2 in patients with breast cancer can be used as a candidate diagnostic indicator and   for diagnos
is of the benign and malignant breast diseases.

Abstract:Objective To observe the vascular casting specimen of medial supramalleolar cutaneous branch of posterior tibial artery，and investigate  the  surgical design of medial supramalleolar cutaneous branch flap of posterior tibial artery for the repairing of leg-foot skin and soft tissue defect，and illuminate the blood supply and characteristics of medial supramalleolar cutaneous branch of posterior tibial artery  to provide anatomical basis for clinical application.Methods Vascular casting and gradation etching were performed on 20 fresh adult cadaver lower legs，series data were obtained with vernier caliper and graph paper;the emerging point，branches，distribution，anastomosis，shape and area of blood supply of medial supramalleolar perforating branch of posterior tibial artery were collected and analyzed．Results In 90%，the emerging point of medial supramalleolar cutaneous branch of posterior tibial artery were concentrated in 8-12 cm of projection distance on condyle-ankle line，the mean diameter of the vessel was （0.96±0.24）mm；anatomical blood supply region was similar to oval，the area was （72±8）cm²，occuping 14.22% of the whole shank.The emerging point of medial supramalleolar cutaneous branch of posterior tibial artery was made as zero point，the line between emerging point and the middle point of posterior border internal malleolus and calcaneal tendon as longitudinal axis，the line through zero and vertical to longitudinal axis as horizontal axis，the blood supply region of medial supramalleolar perforating branch of posterior tibial artery was divided into four quadrants，the blood supply areas of every quadrant were upper inner quadrant（15±2）cm²,upper outer quadrant（8±3 ）cm²，lower inner quadrant（25±4）cm²，lower outer quadrant （24±3） cm²；the second class branch and third class branch were distributed more in upper inner and lower outer quadrants，the axial direction (Dmax/L) was 1.47±0.31；the cutaneous branch had abundant vascular anastomosis with vessels nearby，and participated in the composition of nutrient vessel chains of saphenus nerve，the effective blood supply region could reach 2/3 of  inferior medial facies of the shank． Conclusion Medial supramalleolar cutaneous branch of posterior tibial artery has relatively invariable emerging point，large blood supply region,and is easy to cut and transpose.According to its branches，distribution and anastomosis conditions，the surgical design of double axis-point and double axis flaps with medial supramalleolar cutaneous branch of posterior tibial artery can be made as feeding artery to repair leg-foot skin and soft tissue defect．

Abstract:Objective To study the relationship between the unbalance of internal environment of blood vessel and obstructive sleep apnea-hypopnea syndrome(OSAHS) compliated with type 2 diabetes and  provide a theoretical basis for protection of the blood vessel  endodermis.Methods 42 patients with OSAHS,38 patients with type 2 diabetes and 32 patients with OSAHS complicated with type  2 diabetes were selected,all the patients  were diagnosed by polysomnography(PSG) and the level of fasting blood sugar.The plasma endothelin(ET-1)and serum nitric oxide(NO) levels were measured by radioimmunoassay and nitrate reductase method respectively.Results The level of plasma ET-1 in patients with OSAHS complicated with type  2 diabetes was higher,the serum NO level and NO/ET-1 were lower than those in the  patients with OSAHS and type 2 diabetes,there were  significant differences (P<0.05).The linear correlation results showed that the plasma ET-1 level was positively  correlated  with the fasting blood sugar(r=0.729),the serum NO level was negatively correlated  with fasting blood sugar(r=－0.586) in the  patients with type 2 diabetes. The plasma ET-1 level was  positively correlated with AHI(r=0.820),the serum NO level was negatively correlated with AHI(r=－0.734) in OSAHS patients.Conclusion The    plasma ET-1 level of patients with OSAHS complicated with type  2 diabetes is higher,the serum NO level and NO/ET-1 are lower than those in  the patients with OSAHS and type 2 diabetes.It demonstrates that the injury of the blood vessel endodermis is more serious  in  patients with OSAHS complicated with type  2 diabetes.

Abstract:Objective To observe the effects of inhaled glucocorticoids on the two kinds of inflammatory bronchial asthma by analyzing sputum inflammatory phenotypes of the asthmatic patients with  eosinophilic asthma (EA) or non-eosinophilic asthma (NEA).Methods 156 cases of patients with asthma and 30 cases of healthy control were enrolled in the study.Induced sputum technique was applied to collect sputum and for cytological analysis,the patients with asthma were devided into EA 86 cases and NEA 70 cases.The patients with asthma were inhaled with fluticasone 500 μg·d-1 and placebo,4 weeks later,the airway responsiveness and life  quality score for asthma were observed.Results In NEA patients,the number of sputum eosinophils was significantly decreased compared with EA patients (0.5%±0.1%  vs  9.9%±0.2%,P<0.05),and the number of neutrophils was higher than that of EA patients(64.5% ± 8.5%  vs  37.3% ± 7.5%,P<0.05). After inhaled fluticasone  500 μg·d-1 for 4 weeks,in EA patients,compared with placebo group,the score of life quality in treatment group was significantly increased(P<0.01),the methacholine PC20 was increased group(P<0.05),and FEV1(%)was also increased (P<0.05),the number of sputum eosinophils was significantly decreased (P<0.01);while compared with placebo group,the above indexes in NEA patients had no significant difference(P>0.05).The differences   of  methacholine PC20,sputum eosinophil numbers and the scores of life quality after treatment between EA group and NEA group were significant (P<0.05) .Conclusion NEA is characterized by the less number of airway eosinophils.After inhaled corticosteroid,the changes of methacholine PC20,sputum eosinophil numbers and the score of life quality in patients are not significant.But the characteristics of EA are opposite.So the patients with asthma can be treated targetely based on asthma phenotype according to sputum analysis.

Abstract:Objective To evaluate the association of 5-hydroxytryptamine 1A receptor gene( 5-HT1A receptor gene) rs6295  polymorphism and suicide with Meta-analysis. Methods PubMed,Cochorane,CNKI and Wanfang Database were used to  search all the relevant case-control trials about the association of 5-HT1A receptor gene rs6295 polymorphism and suicide. Statistical analysis was performed with RevMan5.0 including the  heterogeneity inspection, effect consoliating and evaluatation on  publication bias. Results A total of 6 articles were included after the analysis.Because suicide in the literature included suicide attempters and suicide completers, 7 independent studies were selected. Overall the results of suicide genotype Meta analysis were listed as follows: OR=1.27，95 %CI=0.93-1.74,P>0.05，suicide alleles were  OR=1.15,95 %CI=0.91-1.45,P>0.05.The Meta analysis results of suicide attempter genotype  were listed as follows: OR=1.12,95 %CI=0.84-1.49,P>0.05;suicide attempter alleles  OR=1.04,95 %CI=0.87-1.24,P>0.05.The Meta analysis  results of suicide completer genotypes were listed as follows: OR=1.53,95 %CI=1.10-2.14,P<0.01,suicide completer alleles were OR=1.57,95 %CI=0.66-3.71，P>0.05.Conclusion The Meta analysis results show that the  5- HT1A receptor gene rs6295 polymorphisms are not associated with suicide attempters but associated with suicide completers.

Abstract:Objective To study the changes of N-terminal pro brain natriuretic peptide(NT-ProBNP) in patients with type 2 diabetes mellitus(T2DM) complicated with  autonomic neuropathy(AN)(T2DAN), and  illustrate the correlation between NT-ProBNP and T2DAN. Methods 40 patients with simple T2DM，35 patients with T2DAN were selected, and 21 healthy subjects were used as control group.  The expressions of  NT-ProBNP in the peripheral blood of the patients and healthy　 subjects were detected with ELISA. Results The levels of NT-ProBNP in T2DAN group were higher than those in  T2DM group (P<0.01) and healthy control group (P<0.01).NT-ProBNP was positively correlated with course, FPG and HｂA1c，HR in horizontal position,change of SBP from supine to standing (r=0.957, P<0.01;r=0.932, P<0.01; r=0.912, r=0.876，P<0.01;r=0.924,P<0.01), but negatively correlated with FC-P,HRV with deep respiration,HRV with postural hange,30:15 ratio,valsalva ratio(r=－0.755, P<0.01; r=－0.766, P<0.01;  r=－0.698, P<0.01; ;r=－0.732, P<0.01;  r=－0.812, P<0.01). Conclusion NT-ProBNP over-expresses and has a clear correlation with  DAN in patients with T2DAN.

Objective
To compare the actual dose of patients who receive the same medical practice by either digital panoramic X-ray unit and general panoramic X-ray unit and give evidence for better selection of oral X-ray examination method.Methods Round sheet lithium fluoride(LiF)  thermoluminescent dosimeters(TLD) were used.The experiment was divided into natural background contrast group,general panoramic X-ray children group,general panoramic X-ray adults group,digital panoramic X-ray children group and digital panoramic X-ray adults group.The dosimeter of natural background radiation was placed at the office of the doctor,the dosimeters of  general panoramic X-ray children group and  general panoramic X-ray adults group were irradiated by different conditions according to the clinical application of panoramic X-ray to children and adults,the dosimeters of digital panoramic X-ray children group and digital panoramic X-ray adults group were irradiated by different conditions according to the clinical application of digital panoramic X-ray to children and adults.The thermoluminescent dosimeter was used to count and calculate the exposure doses in various groups.Results The dose of children exposed in general panoramic X-ray unit was 1.28 times of that in digital panoramic X-ray unit,there was significant difference（t=6.904,P<0.01）.The dose of adults exposed in general panoramic X-ray unit was 1.55 times of that in the digital panoramic X-ray unit,there also was significant difference（t=－11.514.P<0.01）.Conclusion The digital panoramic X-ray unit can reduce the dose of patients,so the digital panoramic X-ray unit should be used as far as possible.

Objective
To reveal the geographical distribution regularity of normal reference value of young men stroke volume in China,and provide scientific basis for making its unified standard.Methods 3 236 cases of normal young men stroke volume from 68 units in 20 provinces of China were collected.Applying the method of related analysis and regression analysis,the correlations between geographical factors (annual sunshine duration,annual sunshine percentage) and the Chinese normal reference value of young men stroke volume were studied.Results The correlation between geographical factors and the normal reference value of Chinese young men stroke volume was quite significant (F=5.265,P=0.003).Applying the method of Backward in SPSS15.0 software,multiple linear regressions between young men stroke volume and geographical factors were founded,one regression equation was inferred: Y^=1.87+0.022 00X₂+1.002X₃+5.988X₇±14.72.In the above equation,Y^ was normal reference value of Chinese young men stroke volume  (mL),X₂ was annual sunshine duration（h）,X₃ was annual average temperature （℃）,X₇ was annual average wind speed (m·s^-1)，14.72 was the value of the residual standard deviation. Conclusion The normal reference value of Chinese young men stroke volume of some area can be reckoned by using the regression equation.

Objective According to the need of large-scale gene cloning and the construction of protein interaction map in human liver proteome project (HLPP),to reconstruct the yeast two-hybrid plasmids pPC86 and pDBLeu,and then set up the Chinese liver cDNA library,and lay the foundation for further study of HLPP.Methods The specific oligonucleotides were designed and synthesized.They were treated to form double-strand DNA fragments with specifc sticky end,and inserted into pPC86 or pDBLeu vector at the appropriate restrictive enzyme sites to form reconstructed plasmids.The reconstructed vector,Chinese normal liver tissue mRNA and Clontech’s SMART cDNA library construction kit were used to construct NpDBLeu-cDNA library.Results Restriction enzyme digestion and DNA sequencing confirmed the insertion sites and correct sequences of reconstructed vectors.The NpPC86 and NpDBLeu were obtained successfully.Titer tests showed the content of constructed NpDBLeu-Chinese liver cDNA library reached 1.93×10⁶ cfu，in which the percentage of recombinant clones was 95.2%.The titer of amplified library reached 109cfu·mL^-1.Conclusion The Chinese normal liver cDNA library is successfully constructed using SMART technique.

Objective  To obtain cholesterol-reducing lactic acid bacteria (LAB) from Tibet kefir,and lay the foundation for physical character and mechanism study of its cholesterol-reducing effect in vitro and in vivo.Methods The cholesterol-reducing ability was tested on 7 strains of lactic acid bacteria isolated from Tibet kefir by the o-phthalaldehyde method.LA1 strain was screened for its better cholesterol-reducing ability,then subjected to physiological and biochemical identification as well as 16S RNA sequencing,and the acid- and oxgall-resistance,growth curve,pH value and antimicrobial activity were studied.Results The strain of LAB (LA1) was obtained,which presented a comparatively high ability of cholesterol reducing.The strain also showed high acid resistance and bile salt tolerance.The time of LA1 entering the logarithm phase of growth and stationary phase of growth was 10 and 18 h after culture.The strain survived for at least 3 h in the MRS broth with pH 1.5 and grew well in the MRS broth containing 0.3% bile salt.The results of bacteria inhibition experiment showed that the separated lactic acid bacterium had remarkable bacteriostasis on the growth of E.coli and Staphylococcus　aureus.The strain was identified to be Lactobacillus acidophilus by 16S rRNA sequencing.Conclusion The isolated LA1 shows probiotic properties of cholesterol reduction,acid and bile tolerance and antimicobial activity.It can be used as primary strain for study on activity in vivo and mechanism.
 

Objective To establish the high performane liqnid chromatography (HPLC)   for determining emodin in Gengnianan capsule，and provide the basis for the quality control of industrial production.Methods The HPLC was developed to determine the emodin in Gengnianan capsule.The developed conditions of HPLC were as follows: the Shimpack VP-ODS C18(150 mm×4.6mm,5 μm), methanol-0.1% phosphoric acid solution（77：23） as mobile phase,the detection wavelength of  254 nm,and the column temperature of 40℃.Results The linear range of emodin was 0.04-0.32 μg,the regression equation was Y=1060+2513102X，r=0.9991.The average recovery was 98.6%,and the RSD was 1.88% (n=6).The RSD in accuracy,stability and repeatability test were 0.86%,2.33% and 1.88%,respectively.Conclusion This method is accurate，simple and reliable,which provide the scientific basis for the quality control of Gengnianan capsule.

Objective
To evaluate the effects of the INGAP-related pentadecapeptide (INGAP-PP) combined with chemical molecules and cytokines on the differentiation of human adipose-derived stem cells (hASCs) into islet-like clusters. Methods The hASCs  were isolated,purified and expanded in vitro and differentiated using a four-step protocol that included TSA,INGAP-PP/Scrambled peptide (Scrambled-P),nicotinamide and exendin-4. By using real-time PCR and immunofluorescence technology,the expression levels of genes related with pancreas development were detected. The insulin and C-peptide released by differentiated cells in response to high/low glucose level were detected by ELISA.Results Undifferentiated hASCs expressed oct3/4,key markers of embryo stem cells and at the end of induction,the Oct3/4 expression level decreased to 1/20 of original. The expression levels of genes involved in early pancreas development(Ck19,nestin,pdx-1,and nkx2.2) were increased in both INGAP-PP/Scrambled peptide cultures significantly. The islet-like clusters were obtained in both INGAP-PP/Scrambled peptide groups. The diameters of clusters in the INGAP-pp group were much closer to native islets,which were about 150-200 μm,whereas those in Scramble-pp group were about 50-100 μm. Immunocy- tochemical analysis showed C-peptide/insulin positive cells in INGAP-pp induced group but not in undifferentiated hMSCs. Conclusion By INGAP-pp protocol, the hASCs could be differentiated into islet-like clusters secreting the insulin. It suggests that the hASCs could be substitution of embryo or bone-marrow derived stem cells for future cell therapy applications.

Objective
To purify the phosphatidylethanolamine-binding protein 4 (PEBP4) , a new member of the phosphatidylethanolamine-binding protein family from  porcine seminal plasma and lay the foundation for further investigation of its function.Methods Sus PEBP4 was purified from approximately 500 mL porcine seminal plasma by ammonium sulfate fractionation, Q-Sepharose column chromatography, matrex Gel Red A column chromatography, SP-XL-Sepharose column chromatography and Mono-S(A) column chromatography (in FPLC).The amino acid sequence of the purified sus PEBP4 was identified by automated N-terminal sequencing of the intact protein.The amino acid sequences of sus PEBP4 were compared with those of human, bovine, dog PEBP4 by bioinformatics.Results The molecular mass of the purified protein was calculated to be 25 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of beta-mercaptoethanol.The overall homology of amino acid sequence between sus, human, bovine and dog  was from 75.2 to 81.8%.Conclusion Sus PEBP4 is purified by the optimal methods from porcine seminal plasma.Its primary structure is defined.Sus PEBP4 has high homology with other organisms.