To investigate the effect of instilled air on lipopolysaccharide (LPS)-induced mouse model of acute lung injury (ALI) and to provide evidence for establishment of more effective intratracheal instillation. Methods Forty-five male C57BL/6J mice were randomly divided into control group (n=15),LPS group (n=15),and LPS+air group (n=15).The mice in LPS and LPS+air groups were instilled with LPS to establish ALI models by exposed intratracheal instillation method.LPS was instilled intratracheally into the lungs of the mice in LPS+air group with  1 mL syringe prefilled with 100  μL air and the mice in control group were not treated.The detection of the biochemical indexes in bronchoalveolar lavage fluid (BALF),cell differential counting in BALF,lung wet/dry weight (W/D) ratio,and the morphological observation of lung tissue  were performed 24 h after  intratracheal instillation.
Results Compared with control group,the activities of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH),the  concentrations of total protein  in BALF,the amounts of total cells and neutrophils in BALF,and lung W/D ratios of the mice in LPS and LPS+air groups  were significantly increased (P<0.05).Compared with LPS group,the activities of ALP and LDH,the concentration of total protein in BALF,the amounts of total cells and neutrophils in BALF,and lung W/D ratio  in LPS+air group　were significantly increased(P<0.05).The  histological observation results of lung tissue showed that there were differ
ent degrees of fluid accumulation,neutrophils infiltration,congestion and hemorrhage of the mice in LPS+air and LPS groups compared with control group;there were more protein-rich fluid,neutrophils and erythrocytes accumulated in alveolar spaces of the mice in LPS+air group compared with LPS group.
Conclusion The instilled air can be used to improve the method of intratracheal instillation and establish the more reliable experimental animal model of ALI.

Abstract:Objective To investigate the effects of antidepressant olanzapine on the apoptosis,death  and cell cycle of  hippocampus neurons of the rats injured by glutamate (Glu),and to  illuminate the mechanism of neuroprotive effect of olanzapine.Methods The primary cultured hippocampal neurons were injured with Glu to set up Glu-injured models.The neurons  were divided into normal  control group,Glu injury group and olanzapine+Glu injury group.The changes of   apoptotic rate,death rate and cell cycle of neurons were detected by FCM,respectively.Results Compared with normal control group,the  apoptotic rate of neurons in the other  groups were increased (P<0.01);compared with Glu injury group,the apoptotic rate  of cells in 10.0 μmol/L olanzapine+Glu injury groupwas decreased(P<0.01).Compared with normal control group,the death rate of neurons was up to 2.9 times of that in Glu injury group(P<0.05);
compared with Glu injury group,the death rates of neurons in 100.0 and 200.0 μmol/L olanzapine+Glu groups were decreased (P<0.05 or P<0.01).Compored with normal control group,the  percentage of neurons in G0/G1 phase in Glu injury group was increased (P<0.01),and the  percentage of neurons in S phase was decreased (P<0.01),but the  percentage of neurons in G2+M phase didn’t change significantly;compared with Glu injury group,the
  percentages of   G0/G1 phase  in 50.0 and 100.0 μmol/L olanzapine+Glu injury groups were decreased to 79.9% and 62.6% of that in Glu injury group
(P<0.05 or P<0.01).and the  percentage of neurons in S phase  were up to 2.5 an 3.8 times of that in Glu injury group (P<0.05 or P<0.01).Conclusion
Olanzapine can prevent the apoptosis and death of hippocampal neurons induced by Glu,and change the cell cycle.

Abstract:Objective
To observe the killing effects of different kinds of radiotherapy methods  and the radiosensitization of cisplatin on lung cancer cells and to explore the role of autophagy  and related mechanisms by in vitro and in vivo experiments.  Methods
Lung cancer cells  A549 were treated with different concentrations (0,1.0，2.5，5.0，10.0，20.0，and 40.0  μmol?L-1) of cisplatin.40 C57BL/6 mice were randomly divided into  control group(n=10),conventional fractionated radiation group(n=10),hyperfractionated radiation  group (n=10),and  combination group (n=10).The tumor-bearing C57BL/6 models were established. MTT assay was used to detect the cell viability;Western blotting method was used to detect the expressions of autoiphagy-relative gene  MAPLC3 and Beclin1  protein of the  mice in various groups;the tumor volumes of the mice were mesured by caliper;the expression levels of MAPLC3Ⅱ,PI3KⅢ,and Beclin1 in tumor tissue were detected by immunohistochemistry.Results With the increasing of cisplatin concentration(>5  μmol?L-1),the cell viability of A549 cells  was decreased significantly; the cell viabilities in  10.0，20.0，and 40.0   μmol?L-1 cisplatin groups were decreased
 compared with control group(0 μmol•L^-1 cisplatin)(P<0.05).Compared with control group,the expression levels of MAPLC3Ⅱ and Beclin1 in A549 cells in the other groups were increased.The tumor volumes were compared,hyperfractionated radiation  group<  combinatin  group< conventional fractionated radiation group<  control group.Compared with control group,the tumor volumes of the mice in the other groups were decreased（P<0.05）.Compared with control group, the expression  levels of MAPLC3Ⅱ,PI3KⅢ, and  Beclin1  in the other groups were increased significantly(P<0.05）,especially in  combination   group.Conclusion The synergistic cytotoxic effect of ciaplatin with irradiation results in a killing effects on lung cancer cells,and autophagy might participate in this process.
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Abstract:Objective
To observe the  expressions of hypoxia inducible factor-1 alpha (HIF-1α),multidrun resistance(MDR1),and B cell lymphoma/leukemia-2(Bcl-2) mRNA and protein  in human ovarian cancer DDP resistant strain SKOV3/DDP after HIF-1α silencing,and to explore the mechanism of the  resistance of  SKOV3/ DDP reversed by  HIF-1α gene silencing.
Methods The ovarian cancer cell strain SKOV3(sensitivity group) and DDP resistance strain SKOV3/DDP (resistance group)were cultured in vitro and parts of the SKOV3/DDP were transfected with HIF-1α interfere plasmid pshRNA-HIF (transfection group) or control plasmid pshRNA-Control (control group).RT-PCR method was used to detect the expression levels of  HIF-1α,MDR1 and Bcl-2 mRNA;Western blotting and immunohistochemisty methods were used to determine the expression levels of HIF-1α,P-gp (MDR1 gene encoding protein),and Bcl-2 protein in  the cells.Results The expression levels of  HIF-1α,MDR1 and Bcl-2 mRNA   in sensitivity  group and transfection group  were significantly lower than those in resistance group (P<0.05) detected by RT-PCR;the expression levels of HIF-1α,MDR1,and Bcl-2 protein in sensitivity group and transfection group were significantly lower　than those in resistance group(P<0.05) determined by Western blotting and immunohistochemistry;the expression levels of MDR1,Bcl-2 mRNA and protein  in sensitivity group and transfection group had no significant difference (P>0.05).The expressions of HIF-1  α and MDR1 and Bcl -2 were closely related（correlation coefficient of mRNA,r=0.908,P=0;r=0.916,P=0；correlation coefficient of protein,r=0.773,P=0.003；r=0.862,P=0）.Conclusion The resistance of  ovarian cancer cell SKOV3 / DDP HIF-1 α gene silencing may be related to the  down-regulation of   MDR1 and Bcl-2 expressions.

Abstract:Objective
To study the induction of cisplatin on apoptosis of human glioma SHG-44 neurospheres,and to explore its mechanism.Methods The SHG-44 neurospheres were cultured and divided into control and cisplatin groups.The cells in  control group were treated with stem cells media,and based on control group,the cells in cisplatin group were treated with 10 mg•L^-1 cisplatin for 24,48 and 72 h.MTT assay was used to evaluate the  inhibitory rate of proliferation of SHG-44 neurospheres;the morphology of the  cells  was observed by inverted microscope;the levels of Bax,Bcl-2,and caspase-3 proteins secreted by SHG-44 neurospheres  were detected by ELISA assay.Results Compared with control group,the  inhibitory rates of SHG-44 neurospheres in cisplatin group were increased  after treated for 24,48 and 72 h(P<0.01).The apoptotic bodies were observed and the number of neuroshperes were reduced in cisplatin group  under inverted microscope.Compared with control group,the levels of Bax and Bcl-2 were decreased,the level of caspase-3  was increased,and the ratio of Bax/Bcl-2  was significantly increased in cisplatin group (P<0.05 or P<0.01).Conclusion Cisplatin could significantly inhibit the growth of SHG-44 neurospheres and induce apoptosis,and its mechanism may be related to  up-regulating the ratio of Bax/Bcl-2 and giving the apoptosis factors the advantage.

Abstract:Objective
To construct the prokaryotic expression vectors of overall length and different domains of RUNX3 and to identify the expressions of their recombinant proteins.Methods The coding sequence of overall length and different domain truncations of RUNX3 were amplified from the pcDNA3.1-myc-RUNX3 plasmid  by PCR method and inserted into glutathione transferase(GST) fusion expression vector  pGEX-4T-2 through EcoRⅠand BamHⅠ restriction enzyme sites.Then they were transformed into E.coli BL21 and the fusion proteins were induced  and identified by SDS-PAGE electrophoresis.Results The vector fragments FL (1 245 bp),ΔRunt(843 bp),Nter(159 bp),Runt(402 bp), and Cter (684 bp) which were  consistent with the expected fragments were obtained after double enzyme digestion of EcoRⅠand BamHⅠ.The SDS-PAGE electrophoresis result showed that the relative molecular mass of GST-RUNX3 truncated fusion proteins of overall length and different domains of  RUNX3 (GST-FL,GST-Runt,GST-Nter and GST-Cter)were 72 000,40 000,32 000, and 51 000,respectively.Conclusion GST-tagged prokaryotic expression vectors overall length and   different truncated regions of RUNX3  are constructed  and their recombinant proteins are expressed successfully.

Abstract:Objective To observe the effect of NDGA on the survival of  amniotic epithelial cells(AECs) in the chemical acellular muscle scaffold after transplanted into injured spinal cord,and to provide experimental basis for the application
of DNGA in spinal cord injury. Methods  Chemical acellular muscle scaffold combined with Dil labeled AECs of 6 pregnant  rats were  used for spinal cord hemisection injury model. 72 adult male rats  were randomly divided into saline group(n=18),20 mg•kg-1NDGA group(n=18),30 mg?kg-1NDGA group(n=18),and 40 mg•kg-1NDGA group (n=18).The rats in saline group were given intraperitoneal injection of saline. The rats in each NDGA  groups were given intraperitoneal injection of NDGA by the  concentration of 20,30,and 40 mg•kg-1,respectively. And 6 rats in  each group  were separately sacrificed on the 1st,2nd,and 4th week  after operation. The number of the survival AECs waas observed under  fluorescence microscope; the  percentage of positive blood vessel area and the number of NG2 positive cells of the rats in various groups  were observed by immunohistochemical method. Results The number of AECs of the rats in various groups was decreased with the prolongation of time.1,2,and 4 weeks after transplantation,the number of AECs  in  30 mg•kg-1NDGA group and 40　mg?kg-1NDGA grou
p was significantly higher than those in  saline group and 20 mg?kg-1NDGA group (P<0.05). 1 week after transplantation,the  percentages of positive blood vessel area of the rats in  30 mg•kg-1NDGA group and 40 mg•kg-1 NDGA group were significantly lower than those in  saline group and 20 mg•kg-1NDGA group (P<0.05). 1 week after transplantation,the number of the NG2 positive cells of the rats  had no significant difference between various  groups (P＞0.05).Conclusion
NDGA is  able to significantly extend the survival of the transplanted AECs in the chemical acellular muscle scaffold after spinal cord injury. NDGA  can inhibit the new blood vessels of the injured spinal cord,and dose not affect the endogenous neural stem cells.

To observe the effect of 5-ALA photodynamic therapy (5-ALA　PDT) in combination with human telomerase reverse transcriptase mRNA antisense oligodeoxynucleotides (hTERT ASODN) on proliferation inhibition and apoptosis of human ovarian carcinoma 3AO cells,and to disscuse the mechanism of the killing effect of the  combination treatment on  human ovarian carcinoma 3AO cells.Methods MTT assay was used to screen he best appropriate combined parameters of different concentrations of 5-ALA and laser doses on proli
feration inhibition of human ovarian cancer 3AO  cells;the expression changes of  hTERT mRNA  were detected by RT-PCR after transfected with hTERT ASODN and sense oligodeoxynucleotides (SODN).Then human ovarian carcinoma 3AO cells were divided into blank control group,hTERT ASODN transfection group,hTERT SODN　transfection group,5-ALA PDT group,hTERT ASODN transfection combined with 5-ALA
PDT group,and  hTERT SODN transfection combined with 5-ALA PDT group.The inhibitory rates of  proliferation of  human ovarian carcinoma 3AO cells  we
re detected by MTT assay;the  apoptotic rates of  human ovarian carcinoma 3AO cells  were conducted with Annexin-V/PI and flow cytometry.Results The inhibitory rate of proliferation  of 5-ALA PDT on human ovarian carcinoma 3AO cells was dependent on the concentrations of photosensitizer and laser doses(P<0.05).The 24 h transfection in human ovarian carcinoma 3AO cells with different concentrations of hTERT ASODN diminished the abundance of hTERT mRNA in a concentration-dependent manner(P<0.05) and hTERT SODN had no effect on the expression of hTERT mRNA(P>0.05).The inhibitory rate of proliferation  of cells in hTERT ASODN transfection combined with 5-ALA PDT (0.50 mmol•L-1 5-ALA,2.50 J•cm-2 laser dose of PDT) group was  significantly increased compared with 5-ALA PDT group and hTERT ASODN transfection group(P<0.05).The apoptotic  rate of human ovarian carcinoma 3AO cells in hTERT ASODN transfection combined  with 5-ALA PDT group was 29.28%,which was significantly higher than those in  hTERT ASODN transfection group(12.79%) and 5-ALA PDT group (21.19%)(P<0.05).
Conclusion The  administration of 5-ALA PDT  combinated with hTERT ASODN display synergistic proliferation inhibition on human ovarian carcinoma 3AO cells,and the mechanism may be related to promoting apoptosis.
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To investigate the effect of ionizing radiation on the expressions of receptor activator for NF-κB(RANK) mRNA and protein in  differentiation process of osteoclasts and to discuss the molecular mechanism of bone injury induced by ionizing radiation.Methods The osteoblasts were differentiated from RAW264.7 cells after treated by 50 μg•L-1 receptor activatorfor NF-κB ligand(RANKL).The RAW264.7 cells were divided into   control group (normal RAW264.7 cells without treatment),RANKL treatment  group(RAW264.7 cells treated with   50 μg•L-1  RANKL), radiation treatment group (RAW264.7 cells exposed to 2 Gy γ-rays) and  radiation combined with RANKL treatment group(RAW264.7 cells treated with both 2 Gy  γ-rays and 50 μg•L-1  RANKL).Tartrate resistant acid phosphatase(TRAP) staining method was used to assess the osteoclast differentiation status;the expression levels of RANK mRNA and protein of  ostoclasts were detected by  PCR method and Western blotting method,respectivly.Results After treated with RANKL for 7 d,the RAW264.7 cells showed TRAP positive result,which indicated the formation of osteoclasts.Compared with control group,the expression levels of  RANK mRNA and protein  of  osteoclast precursors in RANKL treatment group and radiation treatment group  were increased (P<0.05).Compared with RANKL treatment group,the expression levels of RANK mRNA  and protein in radiation  combined  with RANKL  treatment group were decreased(P<0.05). Conclusion Ionizing radiation  may promote the proliferation and mature,and increase the activity of osteoclast precursors,but it  can inhibit the proliferation,mature,and activity of osteoclasts.

To compare the differences of effects between   four kinds methods  on setting t
he optimal positive end expiratory pressure （PEEP）in animal models  with acute respiratory distress syndrome(ARDS),and to clarify the  feasibility of these methods to  confirm the optimal PEPP and to provide basis for ARDS treatment in clinic.
Methods The ARDS pig models were induced by intravenously infusing oleic acid(n=7).The optimal PEEP  level was determined by four kinds of  methods after lun
g recruitment:such as optimal PaO2 + PaCO2 method,optimal oxygenation method,optimal static compliance（Cst） method,and optimal dynamic compliance (Cdyn) method. Intrapulmonary shunt fraction（Qs/Qt）,plateau airway pressure (Pplat),Cst,dynamic compliance(Cdyn),oxygen index（OI）,central venous pressure（CVP）,cardiac output(CO), global end-diastolic volume(GEDV), intra-thoracic blood volume(ITBV),extravascular lung water（EVLW） of ARDS pigs were recorded before infusing oleic acid，after s
tabilization of ARDS model and at optimal  PEEP level. Results The optimal PEEP
（cmH2O） eleterminated by  optimal PaO2 + PaCO2  method(13.14±1.35),optimal oxygenation method( 13.43±1.51),optimal Cst method( 14.43±4.12 )and  optimal cdyn method(14.14±2.91) had no significant differences (P>0.05).The values of OI,Cst,Cdyn and arterial oxygen saturation(SaO2) in ARDS model were  significantly lower than those at the baseline level (P<0.05).Compared with ARDS model，the values of OI,Cst,Cdyn and SaO2 were significantly improved after using optimal  PEEP(P<0.05).The value of Qs/Qt was increased in   ARDS model compared with the baseline level(P<0.05);and the value of
SaO2 was decreased(P<0.05).The values of Qs/Qt and SaO2 were   decreased to the baseline level after using optimal PEEP.The values of CO,GEDV,and ITBV showed no significant differences(P>0.05) between the baseline level，ARDS model and optimal PEEP level；the value of EVLW in ARDS m
odel was significantly higher than that at the baseline level (P<0.05)，but the optimal PEEP did not change EVLW. Conclusion The four kinds of methods can  identify the optimal PEEP.The optimal PEEP can improve lung compliance and oxygenation，and decrease the intrapulmonary shunt，but has no effect on heart function.

To investigate the effect of ginseng protein (GP) on neurons damaged  by beta amyloid protein1-40 (Aβ1-40) and hydrogenperoxi
de (H2O2),and to clarify the neuroprotective effect of GP and its mechanism. Methods The primary neurons were isolated and purified from cerebral cortex of suckling mouse and  cultured for 12 d,and damaged by 30  μmol•L-1Aβ1-40 and 200 μmol•L-1 H2O2. Then the damaged cells  were randomly divided into model group (Aβ1-40 model group and H2O2 model group,no drugs),GP group and  GP medicated  serum  group. MTT method was used to detect the  survival rates of cells;Qiagen was used to detect the  activities of NOS and levels of NO  in culture supernatant,and  Hoechst 33342/PI fluorescent staining method was used to detect the apoptotic  and necrotic rates of cells. Results It was observed that by the treatment of Aβ1-40  and H2O2,the  number of neuron was decreased,the axons shortened or disappeared,the karyopyknosis concentrated into a bright blue（dyed by Hoechst 33342）,and some cells were dyed into red by PI under inverted flurescence microscope; after adding GP and its medicated serum,the  number of cells was obviously increased,the cells stretched,the  nuclei　were round and light blue,and the number of cells dyed by PI into red was reduced. The MTT results showed that the  survial rate of cells in   GP group was obviously higher than that in model group (P<0.05 or P<0.01).There was no significant difference in  the  survival rate of cells between  GP medicated serum group and  model group (except 5% GP medicated serum group,P>0.05). The activity of NOS and level of NO in culture supernatant in  GP group were significantly lower than those in model group (P<0.05 or P<0.01). The apoptotic rates of cells in GP group and GP medicated serum group were obviously lower than that in model group detected by Hoechst 33342/PI method(P<0.05 or P<0.01). There was no significant differences of  the necrotic rates of neurons between GP group, GP medicated group and model group(P>0.05).Conclusion GP protein has the neuroprotective effect,and its mechanism is related to the reduction of the activity of NOS and the level of NO and the inhibition of apoptosis of neurons.

To discuss the protective effect of different concentrations of Bilberry anthocyanidin on microwave radiation damage of vascular endothelial cells and to clarify the application value of anthocyanidin in radiation injury.Methods The anthocyanidin was extracted by the method of microwave-assisted.The EVC 304 cells were cultivated in vitro.The cells were divided into control group,20 mW•cm-2 microwave radiation group and 25,50,and 100 mg•L-1 anthocyanidin group.The changes of the cell morphology were observed by inverted microscope,the inhibitory rate of proliferation was detected by MTT method,the cell cycle and apoptotic rate were detected by flow cytometry,and the activity of caspase-3 was assayed.Results The cell morphology in microwave radiation group showed contraction,rounded,smaller size,cell gap widening,and most of the cells showed breaking phenomenon.Compared with microwave radiation group,the morphology of cells  in  anthocyanidin group was significantly improved.The MTT experiment result showed the inhibitory rate of  proliferation of EVC 304 cells reached 58.6% after  20 mW?cm-2 microwave radiation for 24 h  and the inhibitory rate of proliferation  of cells in anthocyanidin group was 30.5%,which was lower than that in  microwave radiation group (P<0.05).The flow cytometry results showed that the apoptotic rate of the  cells  in control group was 2.9%;25,50,and 100 mg?L-1 anthocyanidin could  reduce the apoptotic rate of the cells in  microwave group from 14.9% to 6.1%,4.2%,and 3.5%.Compared with  microwave radiation group,the activity of caspase-3 of the cells in microwave radiation group was higher than that in control group(P<0.05). 25,50, and 100 mg?L-1 anthocyanidin could reduce the activity of caspaase-3,which was lower than that microwave radiation group (P<0.05).Conclusion Bilberry anthocyanidin has protective effect on
the radiation damage of vascular endothelial cells.

Abstract:Objective To clone the  glucosidase gene from Thermotoga neapolitana (DSM 4359) and express it in  Escherichia coli,and to study the characteristics of the recombinant enzyme. Methods Based on the glucosidase gene  DNA of Thermotoga neapolitana (DSM 4359),the primers were designed.The glucosidase gene was obtained by PCR method,and the  glucosidase gene was inserted into the expression vector pET-28a(+)and transformed into E.coli BL21(DE3).The glucosidase gene was induced by IPTG and   expressed in a high efficiency in vitro.The optimum reaction temperature,pH value,and the specificity of substrate of the   recombinant  enzyme were observed.Results 825 bp DNA fragment was obtained by PCR amplification,which encoded 274 amino the acid residues,and the  glucosidase gene was inserted into the expression vector pET-28a(+).The recombinant  glucosidase was successfully transferred into  E.coli BL21(DE3) and highly expressed in vitro.The recombinant protein was obtained and had a relative molecular mass of 63 000  indicated by non-denaturing polyacrylamide gel electrophoresis analysis.The optimum reaction temperature of glucosidase was 75℃,and the activity  maintained 60% with the temperature in the range of 70℃-85℃.And the optimum pH value was 5.0,and the activity  maintained 70% with the pH value in the  range of 3.0-6.0.The preferred substrate was trehalose,followed by  gentiobiose,and the  activities  of the other  disaccharides  were  lower or zero.Conclusion Glucosidase gene from Thermotoga neapolitana is successfully cloned and highly expressed in E.coli BL21 (DE3 ).The glucosidase can specifically catalyze the hydrolyzed hydrolysis of α,α-1,1-glucosidic bond.

Abstract:Objective To explore the  effects of irbesartan on exrpession of periostin in  myocardium  tissue of  the viral myocarditis (VMC) mice  in chronic phase and myocardial fibrosis,and to clarify the possible mechanism of its antimyocardial fibrosis.Methods The models of VMC in chronic phase were founded by injecting coxsackie virus B3(CVB3) solution into 40 BALA/c mice. 21 VMC mice were divided into model group（n=11）and irbesartan group（n=10）.And at the same time, 10 BALA/C mice were used as control group.The mice in model group and control group were fed with water (1 mL?d-1),and the mice in irbesartan group were fed with irbesartan at a dose of 50 mg?kg-1?d-1.After 60 d,the levels of left ventricular end-diastolic dimension (LVEDd),left ventricular end-systolic dimension (LVEDs) and left ventricular fractional shortening (FS) of the mice in 3 groups were examined;the concentrations  of AngⅡ in the serum of the mice in various groups were detected by ELISA method;the myocardium tissue of mice were stained by MASSON and cardiac collagen volume fraction(CVF) was accounted.At the same time,the contents of periostin mRNA and TGF-β1 mRNA in myocardium tissue of mice were detected by RT-PCR method.Results Compared with control group,the levels of  LVEDd and LVEDs were  increased （P<0.01），and the level of FS was decreased （P<0.01） in model group;the levels of serum Ang Ⅱ,CVF,TGF-β1 mRNA,and periostin mRNA were increased （P<0.01）in model group.Compared with model group,the levels of LVEDd  and  LVEDs of the mice in irbesartan group were obviously decreased（P<0.05），and the level of FS was increased（P<0.05）.There was no significant difference of the levels of serum Ang Ⅱ between irbesartan group and model group（P>0.05),but the levels of CVF,TGF-β1 mRNA,and periostin mRNA of the mice in irbesartan group were obviously decreased compared with model group（P<0.05）.Conclusion Irbesartan can offer some protection to myocardial fibrosis inVMC mice. The  probable mechanisms may be acted through  inhibiting  the expression of TGF-β1 in myocardium tissue to reduce the expression of  periostin.

Abstract:Objective To explore the effects of atorvastatin  pretreatment (AT) on the expressions of HIF-α and VEGF protein and mRNA at different time points in myocardium tissue of the  rats after myocardium ischemia reperfusion(I/R) and to elucidate the protective effect of AT  on myocardium tissue of the rats.Methods 50 healthy male Wistar rats were randomly divided into sham operation group,ischemia 30 min reperfusion 2 h model (I30minR2h) group,ischemia 30 min reperfusion 6 h model  (I30minR6h) group,ischemia 30 min reperfusion 2 h atorvastatin (I30minR2hAT,10 mg/kg) group, and ischemia 30 min reperfusion 6 h atorvastatin (I30minR6hAT,10 mg/kg) group;there were 10 rats in each group. I/R model was prepared by ligating the left anterior descending coronary artery(LAD) method.The ratios of mypcordial infarction area of the  rats in various groups were detected,the expressions of HIF-α and VEGF protein were detected with immunohistochemical method in myocardium tissue of the  rats,the expression levels of HIF-α and VEGF mRNA in myocardium tissue of the  rats were detected by  RT-PCR method. Results Compared with sham operation group,the ratios of myocardial infarction area of the rats in I30minR2h group and I30minR6h group were increased(P<0.01);the ratios of myocardial infarction area of the rats in I30minR2h AT group and I30min R6h group were lower than those in I30minR2h group and I30minR6h group(P<0.01).Compared with sham operation group,the expression levels of HIF-α and VEGF protein and HIF-α and VEGF mRNA in I30minR2h group,I30minR6h group,I30min R2h AT group,and  I30min R6h AT group were significantly increased (P<0.01);the expression levels of  HIF-α and VEGF protein  and HIF-α and VEGF mRNA  of the  rats in I30minR6h group  were lower than  those in  I30minR2h group,and the expression levels of HIF-α and VEGF protein  and HIF-α and VEGF mRNA of the rats in I30minR6h AT group  were lower than  those in  I30minR2h AT group(P<0.01).Compared with I30minR2h group,the expression levels  of HIF-α and VEGF protein  and HIF-α and VEGF mRNA of the rats  in I30minR2h AT group were significantly increased(P<0.01);compared with I30minR6h group,the expression levels of HIF-α and VEGF protein  and HIF-α and VEGF mRNA of the rats in I30minR6h AT group were significantly  increased (P<0.01). Conclusion I/R injury  can  early cause up-regulation of the levels of HIF-α and VEGF protein and HIF-α and VEGF mRNA in rats,and AT  can significantly up-regulate the expression levels of HIF-α and VEGF protein and HIF-α and VEGF mRNA,which suggests that AT  has the protective effect on the myocardium tissue of rats.

Abstract:Objective To establish doxorubicin　(DOX)-induced myocardial injury rat model and to explore the protective effect of the pseudo-ginsenoside GQ (PGQ) on DOX-induced myocardial injury in rats. Methods The  acute myocardial injury  model was established through the femoral vein injection of doxorubicin,and 30 Wistar male rats were randomly divided into  control group(n=5),DOX group(18 mg?kg-1,n=5),DOX+dexrazoxane(DZR) group(180 mg/kg,n=5);DOX+low,medium,high doses of  PGQ groups(3,6,12 mg/kg/d,n=5). The general state,the heart weight/body weight(HW/BW) ratio, the  cardiac function indexes,the levels of  serum troponin(TnI) of the rats in various groups were detected and the  morphological and ultrastructural changes of myocardium tissue of the rats were observed. Results Compared with  control group,the total decline value of BW of the rats in DOX group was significantly increased(P<0.05);HW/BW ratio was  significantly increased(P<0.05); heart rate(HR),left ventricular systolic(LVSP) and ±dp/dtmax were  significantly decreased(P<0.05),and the level of serum TnI was significantly increased(P<0.05).The morphological and ultrastructural injuries of the myocardium tissue were observed. Compared with DOX group,the HR,LVSP,±dp/dtmax  of the rats in DOX+DZR group and DOX+medium dose of PGQ group were significantly increased(P<0.05),and the levels of serum TnI were significantly decreased(P<0.05),and HW/BW ratios were  significantly decreased(P<0.05).The morphological and ultrastructural injuries were significantly improved compared with DOX group. Compared with DOX+DZR group,the total decline value of BW of the  rats in  DOX+medium dose of PGQ group was significantly decreased(P<0.05),and the  ultrastructural injury was significantly improved.Conclusion  PGQ has the protective effect of myocardial injury against DOX-induced myocardial injury.

Abstract:Objective To observe the changes of the levels of serum sex hormones in mice treated with atrazine (ATR), and to evaluate the effect of  ATR on sex hormone related diseases. Methods 32 healthy female C57 mice were randomly divided into control group and 5, 25,and 125 mg/kg ATR groups;there were 8 mice in  each group.The mice in control group were given olive oil by gavage,and the mice in the other groups were given 5,25, and 125 mg/kg ATR once per day for 28 d. The serum of mice  were collected. The levels  of serum sex hormones  were detected by radioimmunoassay,and HE staining was used to observe the morphology of ovarian tissue of the mice in each group. Results Compared with control group,the levels of estradiol (E2) and testosterone (T) were significantly increased(P<0.05) in 25 and 125 mg/kg ATR groups, and the levels of E2 and T  were  increased with the increasing of ATR dose.Compared with control group,the levels of  luteinizing hormone (LH) and follicle stimulating hormone (FSH)  were significantly decreased (P<0.05 or P<0.01) in 25 and 125 mg/kg groups, especially in  125 mg/kg group (P<0.01).The levels of progesterone (P) and prolactin (PRL) had no significant differences between each group(P>0.05). Compared with control group,the proportions of the development follicles were  reduced in 25 and 125 mg/kg ATR groups, and the proportions of the large atretic follicles were increased,especially in  125 mg/kg ATR group. Conclusion ATR may increase the levels of serum E2 and T  and decrease the  secretion of  LH and FSH in order to interfer  of the ovarian follices.

Abstract:Objective To study the influence of schisandrae water extract in the oxidative stress injury of liver cells of the  rats induced by   microwave radiation, and to clarify the preventive effect of schisandrae water extract on hepatic injury induced by microwave radiation in  rats.Methods 36 normal adult Wistar rats were randomly divided into normal control group(n=12),microwave radiation group(n=12),and schisandrae water extract prevention group(n=12).The rats in schisandrae water extract prevention group were pretreated individually with schisandrae water extract for 5 d before  microwave radiation,after  microwave radiation and schisandrae water extract treatment,the concentration of MDA and the activity of  SOD of the rats 0 and 16 h after radation in various  groups were detected,and the morphological changes of the liver tissue and liver cells were  observed by HE staining.Results The concentration of MDA and the activity of SOD of the rats in microwave radiation group had no significant difference  compared with  normal control group 0 h after microwave radiation.16 h after   microwave radiation,the concentration of  MDA was increased(P<0.05),and the activity of SOD of the  rats in  microwave radiation group was obviously decreased(P<0.05).The concentration  of MDA was decreased and the activity of SOD was increased in schisandrae water extract prevention group (P<0.05). Microwave radiation casused obvious edema of the liver cells,but the pathological injury of the liver tissue  in the rats in schisandrae water extract prevention group was improved.Conclusion 200 mW?cm－2 microwave radiation can induce the changes of oxidative stress level and morphology of liver cells of the rats,and   schisandrae water extract shows obvious therapeutical effects on the  stress injury of liver cells in Wistar rats induced by microwave radiation.

Abstract:Objective To observe the influence of  carvedilol in myocardial fibrosis in rats with heart failure after myocardial infarction and to explore the  mechanism of protective effect of carvedilol. Methods The left anterior descending coronary arteries of Wistar rats were  ligated to establish  rat models of heart failure. 32 rat heart failure models were established successfully after 6 weeks.32 rats with heart failure were divided into sham operation group(n=8),model group(n=8),model+low dose of carvedilol group(n=8),and model+high dose of carvediol group(n=8).The rats in model+low or high doses of carvedilol groups were  treated with carvediol in  low or high  doses (1 and 10 mg/kg carvediol) for 7 weeks  after the operation.The rats in sham operation and model groups were treated with 0.5% CMC-Na 10 mL?kg-1?d-1.All treatment regimens were initiated 6 weeks after operation and continued for 5 weeks.The activity of serum superoxide dismutase(SOD) was detected by xanthine oxidase method and the concentration  of serum malondialdehyde(MDA) was  detected by thiobarbituric acid method; the level  of serum BNP was detected by ELISA method;after Masson staining,the myocardial fibrosis of these rats were observed under light microscope.Results Compared with sham operation group,the serum BNP level and MDA concentration were increased(P<0.05) and the activity of SOD was decreased of the rats in the other groups; compared with  model group,the concentrations  of  serum MDA were decreased  and the activities of SOD were increased  significantly of the  in rats in model +low or high doses of carvedilol groups (P<0.01); the degree of myocardial fibrosis was reduced.Compared with model + low dose of carvedilol group,the serum BNP level was decreased(P<0.05),the concentration of  MDA  was decreased (P<0.05),and the SOD activity  was increased (P<0.05) of the  rats in  model + high dose of  carvedilol　group; the degree of  myocardial fibrosis was reduced.Conclusion Different doses  of  carvedilol can protect the myocardium  tissue and reduce the degree of  myocardial infarction;its mechanism may be related to  decreasing the concentration of serum MDA and  increasing the activity of serum SOD of the rats with heart failure.

Abstract:Objective To investigate the expression and function of intravitreal injection of pigment epithelium-derived factor (PEDF) gene eukaryotic expression vector in proliferative vitreoretinopath (PVR) model eye,and to clarify the inhibitory effect of PEDF gene transfection on PVR  induced by macrophages in rats. Methods The pcDNA3-PEDF with cationic liposome was injected into vitreous of 36 SD rats (the right eye of each rat as the experimental eye and the left eye as the negative control eye).The rats were divided into PVR control group (injecting macrophages in the vitreous cavity to induce PVR),saline control group (injecting macrophages 3 d after intravitreal injection of saline) and transfection group (injecting macrophages 3 d after intravitreal injection of liposome and  pcDNA3-PEDF),and there were 12 rats in each group.Then the ophthalmoscope and fundus pre-set lens were used to observe the vitreous and retinal conditions of the rats in each group;the stuctures of the  vitreous and retinal layers and the cell proliferation of the specimens of the  rats in various groups were detected by HE staining  under light microscope.Results The ophthalmoscope and fundus pre-set lens results showed that  after intravitreal injection of macrophages,there was  more significant vitreous proliferation in PVR control group and saline control group compared with transfection group 1 week after injection,there was vitreous proliferation in PVR control group and saline control group.2-3 weeks after injection,the proliferation was gradually increased and retina began to detach in PVR　control group and saline control group.4 weeks after injection,there was significant proliferation in all model eyes,and traction retinal detachment occurred in PVR control group and saline control group.Only one   eye of the rats in transfection group showed vitreous traction and shallow retinal detachment,and the other eyes  were normal.The HE staining and light microscope results showed that  the structurs of retina cells of most eyes  were  clear,only one eye occured shallow retinal detachment  with epiretinal membrane proliferation and  small amount of inflammatory cell infiltration in transfection group.There was inflammatory response with the progression of the disease,accumulation of inflammatory cells,proliferation of fibroblasts, and  formation of eyeball atrophy in PVR control group and saline control group. Conclusion PEDF transfected by intravitreal injection in rats’ eyes
can significantly inhibit the  formation and development of PVR induced by macro
phages.

Abstract:Objective To investigate the cellular proliferation  level in the rostral migratory stream (RMS) of the adult brain in mice expressed human α-synuclein  (SNCA) A30P mutation and to explore the possible effect of SNCA A30P mutation  on  the neurogenesis in adult brain of mice. Methods 12 mice were divided into C57BL group (normal mice),SNCA-/- group (knocking out SNCA mice), and A30P SNCA-/- group (knocking out  SNCA and knocking in SNCA  A30P mice).The  mouse brains were taken out and the coronal sections were made.The  morphologies of RMS of the mice in various groups were  observed;after the free-floating fluorescent immunohistochemistry of phosphohistone 3(PH3),doublecortin(Dcx), and 4′,6-diamidino-2-phenylindole dihydrochloride(DAPI) were carried out,the number of PH3 positive cells and the fluorescence intensity of Dcx-positive neuroblasts were analyzed.Results There were no signifrcant differences of the morphology,size and cell density of mouse RMS between various groups.Compared with normal control group,the number of PH3 positive  proliferating neurons in RMS of mice in  SNCA-/- and A30P SNCA-/- groups were decreased(P<0.05);compared with normal  control group,the fluorescence intensities of Dcx-positive  neuroblasts in the other two  groups were increased slighthy.Conclusion Human SNCA A30P point mutation may  decrease the neuron proliferation,but slightly enhance its migration in RMS of adult brain of mice,which suggests that the mutation has a certain inhibitory  effect on the neurogenesis in  RMS of adult brain.

Abstract:Objective  To analyze the influence of cyclooxygenase-2(COX-2) inhibitor NS-398 on cell morphology,cell cycle,proliferation and apoptosis of hepatocelluar carcinoma cells SMMC-7721,and to investigate the mechanism of the effect of NS-398   on the growth of tumor cells. Methods The SMMC-7721 cells were treated with different doses of NS-398 (0,25,50,75,100,and 150 μmol/L) in various groups  for 24,48 and 72 h,and at the same time normal control group was set up.The changes of cell morphology in various groups were observed,the inhibitory rates  of proliferation  of hepatocellular carcinoma cells were detected by MTT assay,and the changes of cell cycle and apoptotic rates  were detected  by flow cytometry. Results After treated with 25,50,75,100,and 150 μmol?L-1 NS-398,the cell adhesion was decreased,and some of the cells were floating,and the medium was not clear.Compared with normal control group,the ratios of SMMC-7721 cells in  G0/G1 phase were reduced,and the ratios of SMMC-7721 cells in  G2/M phase were increased in the other groups.No apoptotic peak was observed in normal control group.The apoptotic peak was observed in 50 μmol?L-1 NS-398 group after treated for 24 h,and  the level of apoptosis was increased significantly  72 h after treatment.The  inhibitory rates of proliferation   had a concentration- and time-dependant trend in various groups.Conclusion The COX-2 inhibitor NS-398 can repress the proliferation  of  hepatocellular carcinoma   cells SMMC-7721 and induce   apoptosis;the process is time- and dose-dependant.

Abstract:Objective To investigate the expressions of tumor necrosis factor-α (TNF-α) in main organs and serum of mouse model infected by Ｍycoplasma pneumonia (MP)  and to clarify the  relationship between TNF-α and MP infection.Methods MP was cultured in the medium containing PPLO broth.Total 72 BABL/C mice were randomly divided into control group(non MP-infected mice，n=24),MP infection group (MP-infected mice，n=24)and  antibiotic treatment group(MP-infected mice treated with azithromycin，n=24).MP infection models were established with the method of dripping cultured MP into nasal cavity of mice.ELISA method was used to assay the levels of TNF-α in the serum of the mice in various groups.Total RNA in the lung tissue of mice was extracted and RT-PCR method was used to detect the expression levels of TNF-α mRNA.Immunohistochemical technique was applied to show the lesion of     lung,liver,kidney,and heart tissues in the MP-infected mice.Results The MP-infected mouse models were established successfully,and MP strains were dripped into the  respiratory system of mice.The activities of the mice in  MP infection group and antibiotic treatment group were reduced,and they showed poor eating,listlessness,nasal secretions,cough,piloerection,chills ect.Contrastly,there were no changes in eating,drinking and activity of the mice in control group.The ELISA results showed that the level of  serum TNF-α of the mice in  MP infection group was higher than those in antibiotic treatment group and control group(P<0.05）.At the same time,the level of serum TNF-α of the mice in antibiotic treatment group was higher than that in control group(P<0.05）.The RT-PCR results  showed the level of TNF-α mRNA in lung tissue of the mice in MP infection group was higher than those in control group and antibiotic treatment group(P<0.05),and  the level of TNF-α mRNA of the mice in MP treatment group was higher than that in  control group(P<0.05).The imunohistochemical  results showed the lesion of   lung tissue  of the mice in  MP infection group was heavier than those in  antibiotic  treatment group and control group,and  it was sligher in antibiotic treatment group  than   control group,while there was no    disease or minor lesion of lung tissue of the mice in control group（P<0.05）. The results from the three tests coincided with each other.Conclusion The positive  expression level of TNF-α is  related to the severity degree of MP infection.

Abstract:Objective
To investigate  the formation and density of vasculogenic minicry(VM)  in colon cancer cells,and   to clarify the relevance between VM and cell migration and invasion,and to provide a new starting point and theoretical basis for clinical treatment of colon carcinoma.Methods The colon cancer cell lines HCT8,HCT116,LS174T and HT29 were cultured in three-dimensional condition in vitro,and the formation of VM was observed under inverted microscope;then the density of VM was calculated;the cell migration was tested by wound healing assay,while the cell invasion was observed by Transwell  assay;the linear correlation analysis was used to detect  the relationship between the  density of VM and tumor cell migration distance or the number of cells penetrating the membrane.Results The   HCT8 and HCT116 cells cultured in three-dimensional condition could form VM  in vitro,and the densities of VM of HCT8 cells were (6.75±0.957) and (5.75±0.957)/ visual field,while the LS174T,HT29 cells were unable to form VM.The results of wound healing assay showed that the migration distances on the picture   of HCT8,HCT116 and LS174T cells were (3.833±0.831),(3.967±0.975), and (0.817±0.333) cm,respectively;but the  HT29 cells had no migration trend.The Transwell assay results showed that the number of HCT8,HCT116,LS174T， and HT29 cells penetrating the membrane of every vision was 71.6±4.506,22.4±1.517,0.6±0.894,and 0.2±0.447,respectively.The density of VM was positively correlated with the migration distance (r=0.934,P<0.05) and the number of cells penetrating the membrane detected by Transwell  assay (r=0.853,P<0.05).Conclusion VM exists in the colon cancer cells,and VM may be contributed to the migration and invasion of colon cancer.

Objective To investigate the effect of lidamycin (LDM) combined with bortezomid (BZM) against multiple myeloma and the mechanism of the combination treatment.

Methods The human multiple myeloma SKO-007 cells in logarithm growth phase were selected and  randomly divided into control group,LDM group,BZM group,and BZM combined with LDM group. MTS assay was used to detect the survival  rate of SKO-007 cells and  flow cytometry  was used to analyze
the distribution of cell cycle and  apoptotic rate of the proliferation cells in various groups;the expression levels of protein associated with apoptosis and endoplasmic reticulum stress (ERS) of SKO-007 cells in various groups were detected by Western blotting method.
Results After  culture for 48 h,the survival  rate of the  cells  in BZM combined with LDM group was lower than
those  in control,LDM and BZM groups (P<0.05). The percentage of cells at  G₂/M phase, the  apoptotic rate  of cells,
 the expression levels  of cleaved caspase-3 and poly ADP-ribose polymerase (PARP) of SKO-007 cells,the expressio
n levels  of GRP78/Bip,CHOP/GADDl53,and phosphorylation of c-Jun NH2-terminal kinase (p-JNK
) in BZM combined with LDM group were higher than those in control,LDM and BZM
 groups (all P<0.05). Conclusion BZM can greatly enhance the efficacy of LDM against
multiple myeloma by increasing the levels of cleaved caspase-3 and PARP,and remarkably increase the apoptosis induced by LDM through further activation of ERS.

Objective  To explore the  effects  of taurine on
the expression of tumor necrosis factor-α(TNF-α) and apoptosis  of
  alveolar macrophages(AM) in silicosis rats and its action mechanism,and to provide basis for
treatment of silicosis. Methods 24 rats were randomly divided into control group, model group and taurine
group,and there were 8 rats  in each group.The AM of the  rats were collected from bronchoalveolar lavage fluid(BALF).
The levels of TNF-α in BALF  in different groups were measured by ELISA method,the apotosis of the AM of the rats was
measured by TUNEL method,and  agarose gel electrophoresis was used to evaluate the alteration of DNA fragment.
Results The levels of  TNF-α of rats in control group,model group and taurine group were (215.8±11.7),(528.2±23.3),
and (364.3±21.4) ng?L-1,respectively.Compared with model group,the TNF-α expression level in BALF of silicosis rats in taurine g
roup was decreased(P<0.05). The apoptotic index in taurine group (9.38%±1.64%)  was significantly higher
than that  in control group (5.01%±1.17%)(P<0.01) and lower than that in model group (14.03%±0.74%)
(P<0.05). The characteristic DNA ladder appeared in model group,which was not found in control group and taurine group.
Conclusion Taurine can effectively inhibit AM apoptosis induced by SiO2 through down-regulating the expression of TNF-α.

Objective  To study the protective effect of extract of buckwheat chaff (EBC mellitus) on myocardial injury in  type 2 diabetes mellitus (T2DM)
 rats and  to disscuss its mechanism.Methods T2DM rat  models were established  by injecting streptozotocin (STZ) and given high-fat diet.
23 T2DM rats were divided into model control group (n=8),EBC treatment group (n=7),and benazepril treatment group (n=8).Another 7 SD rats were selected as normal control group.The rats in EBC treatment group and benazepril treatment group were administered with EBC （200 mg/kg-1/d-1）
and benazepril （4 mg /g-1/d-1）orally.While the rats in other two groups were administered with　normal water (10 mL/kg-1/d-1) instead.The levels of fasting blood glucose (FBG),body weight (BW),cardiac index (HWI),heart rate (HR),left ventricular systolic blood pressure(LVSP),left ventricular dia stolic pressure(LVEDP) and ±dp/dtmax
　of  rats were measured 8 weeks after treatment  in various  groups.The  morphological  changes of myocardium tissue of  rats were observed with light
microscope.Results Compared with normal control group,the FBG,HWI,LVEDP of the rats  in model group were increased(P<0.01), and the BW,LVSP,HR,and ±dp/dtmax in model control groups were decreased(P<0.05 or P<0.01).Compared with model control group,the BW,LVSP,HR,and ±dp/dtmax were i
ncreased and the  HWI,FBG and LVEDP  of the rats in EBC and benazepril treatment groups were decreased(P<0.05 or P<0.01).The morphology of myocardium tissue
of  the rats in EBC treatment and benazpril treatment groups were improved.Conclusion EBC can improve the histomorphology  of myocardium tissue of the T2DM rats,and its mechanism may be related to up-regulating  the  LVSP,HR and ±dp/dtmax and down-regulating  the  LVEDP.


Objective To investigate the effect of leptin pretreatment on myocardial ischemic reperfusion injury(MIRI) in type 2 diabetic(T2DM) rats,and to ex
plain the mechanism of the protective effect.Methods Sixty healthy SD rats were randomly divided
 into normal control group (n=10) and diabetes mellitus group (DM,n=50),the rats were fed  with basic
 diet and high fat/high sucrose diet for 4 weeks，respectively.The rats in DM group were injected with strepto  zotocin intrapetioneally to

construct DM model.40 DM rats were randomly divided into DM sham operation group (n=10),and DM-MIRI group (n=10), DM-leptin+MIRI group
(n=10),and DM-MIRI+leptin group (n=10).The blood and heart samples were obtained from the rats  with
 empty stomach in normal control group and 3 h  after reperfusion in DM group.The levels of serum tumor necrosisfactor-alpha(TNF-α),

malondialdehyde(MDA),superoxide dismutase(SOD),fasting blood glucose(FBG),leptin, and homeostasis model assessment-IR (HOMA-IR) of the rats in various groups were detected.Results Compared with normal control group,the levels of FBG,FINS,TNF-α,MDA,
leptin,and HOMA-IR of the  rats in DM group were increased,and the activities of SOD were decreased(P<0.01).Compared
with DM sham operation group,the levels of FBG,FINS,TNF-α,MDA,leptin,and HOMA-IR of the rats in DM-MIRI,DM-leptin+MIRI and DM-MIRI+leptin group were significantly increased,and the activities of SOD were decreased (P<0.01).Compared with DM-MIRI group,the levels of FBG,FINS,TNF-α,MDA,leptin,and HOMA-IR of the rats in

DM-leptin+MIRI group  were significantly decreased,and the activities of SOD were increased (P<0.01).
The myocardium tissue of rats in DM-MIRI  group had serious injuries,and the  myocardial cells  were hypertrophy and deformation,and the myocardium tissue was partly necrosis and hemorrhage,and the myocardial fibers were derangement,and the myocardial cells  were visible and ar
ranged loosely and disorderly with infiltration of inflammatory cells of rats in mesenchyme;the myocardial cells of the rats in DM-leptin+MIRI group were still hypertrophy and deformation,and the myocardial structure was disorder,however the hemorrhage,necrosis,and neutrophils aggregation of myocardial tissue were significantly reduced,and the lesions improved significantly;while the pathological changes of myocardial cells of  the  rats in DM-MIRI+lept
in group were less than  DM-leptin+MIRI group.Conclusion Leptin pretreatment may protect the myocardium tissue and  alleviate the MIRI in DM rats,which may be
related to  the improvement of insulin resistance (IR).

Objective To  study the effects of rapamycin(RAPA) and splenectomy on
 the  expression levels of transforming growth factor-β(TGF-β1) in liver and
 spleen tissues of rats  with liver fibrosis and the pathological morphology of
 
  liver and spleen tissues,and to disscuss  its role in the treatment of
 liver fibrosis.Methods  30 rats were randomly divided into normal control group,

RAPA group,liver  fibrosis model group,splenectomy group, and combination treatment
 group; there were 6 rats in  each group.Carbon tetrachloride (CCl4) peanut oil was used to 

establish the  liver fibrosis  rat  models.The  morphological changes of liver and spleen tissues of the  rats in various  grou
ps were observed under light microscope;the  expression levels of TGF-β1  in liver  and spleen tissues and the expressi
on level of  serm TGF-β1 were detected.Results The morphology  results showed that the 
 rats in  liver fibrosis model group presented  mixing drops of fatty degenerati
on,and the inflammatory cells invasion,and small bile duct hyperplasia,and there
 were hyperplasia of fibrous septum wrapping liver cells; the pathomorphological changes of the
 
 rats in RAPA  group,splenectomy  group and combination treatment  group  were  improved with different
 degrees,especially  in combination treatment group.Compared with  liver fibrosis model  group,the expression levels of TGF-β1 of the rats in

splenectomy group,combination  treeatment  group, and RAPA group were decreased (P<0.05 or P<0.01).Compared with liver fibrosis model group,the

expression level of  TGF-β1 in spleen tissue of the rats in  RAPA group was decreased(P<0.01).Compared with normal control group,the expression levels of TGF-β1 in spleen tissue of the rats in liver fibrosis group and RAPA group were increased(P<0.01).At the 8th week,compared with  liver fibrosis model group,the expression lev
els of serum TGF-β1 of the rats in RAPA group,splenectomy group, and combination treatment group were decreased(P<0.05 or P<0.01).Compared with splen
ectomy group,the expression level of serum TGF-β1 of the rats in combination treatment group was decreased(P<0.05),

but there was no significant difference of the expression levels of serum TGF-β1 between combination treatment
 group and RAPA group(P>0.05).Conclusion RAPA and splenectomy can significantly lower the
 expression level of serum  TGF-β 1 and  the expression level of TGF-β1 in liver  and spleen tissues of the rats
 and reduce the morphological changes in the liver tissue and  ease the process of liver fibrosis  in rats.


Objective To analyze the effectiveness and safety of escitalopram and citalopram in the treatment of  depression with Meta-analysis and to provide
reference for the application in clinic of escitalopram and citalopram.Methods Computer　search was performed using
 the Cochrane Central Register of Controlled Trial(Issue 3,2012)，Embase(1966 to 2012.9),PubMed(1948 to 2012.9),Ovid-medline(1966 to 2012.9),VIP（1989 to 2
012.9），CNKI(1979 to 2012.9),and WanFang(1981 to 2012.9),and the randomized controlled
trial (RCT) were collected,then the retrieved studies according to  inclusion and exclusion criteria were screened,and the qualities of included studies were evaluated and RevMan 5.2.1 software was used for Meta-analysis. Results  A total of 195 articles were found and 12 of which were finally included.There
 was no significant difference of effectiveness rates between escitalopram and citalopram in the treatment of depression［(χ2=5.82，df=11，
P=0.88，Z=0.93(P＝0.35）；95%CI （0.87，1.50）］.The incidence of adverse reaction of escitalopram was similar as that of citalopram［χ2=11.18，df=11，P=0.43，Z=0.48(P=0.63）；95%CI （0.75,1.19）］.Conclusion Escitalopram has the similar effectiveness and the incidence of adver
se reaction with citalopram in the treatment of depression.

Objective To investigate the expressions of Livin and cytochrome C( Cyt-C) in tongue squamous cell carcinoma (TSCC) tissue and to
 explore the correlation between their expression levels and  the   clinicaopathological characteristics of TSCC and the correlation between expressions of Livin an
d Cyt-C protein in TSCC tissue. Methods The expressions of Livin and Cyt-C protein were examined by immunohistochemistry SP method in 48 cases of TSCC and 10 casesof normal tongue tissue,and  the relationship between the expressions of Livin,Cyt-C protein and the   clinicopathological characteristics,such as gender,age,histological type and lymph node metastasis were analyzed. Results Livin mainly expressed   in the cytoplasm. The pos
itive expression rates of  Livin protein in TSCC and normal tongue tissue were 66.67% and 0%,respectively,and there was significant difference  (P<0.01).The  high
 
expression of Livin was correlated to the histological differentiation degree of TSCC (P<0.05),and also closely correlated to the cervical lymph node metastasis (P<0.05). Cyt-C  mainly expressed   in the cytoplasm. The positive expression rates of Cyt-C protein in TSCC and normal tongue tissue were 70.83% and 10.00%,respectively,and there was  significant difference (P<0.01).The  expression of Cyt-C was correlated to the histological differentiation of TSCC (P<0.05).The Spearman analysis results showed that  there was a positive correlation between the expressions of Livin and Cyt-C protein  in TSCC tissue(r=0.553,P<0.001). Conclusion There is a positive correlation between the expressions of Livin and Cyt-C protein in TSCC tissue. Combined detection of livin and Cyt-C will help to judge the  malignant degre
e of TSCC and evaluate the  prognosis.

Objective To investigate the  association between the polymorphisms of 4 Tag SNPs of steroid receptor co-activator 1（SRC-1）gene and type 2 d
iabetes mellitus（T2DM） in  Han population from North China,and to discuss whether the SRC-1 g
ene was the predisposing gene of  T2DM.Methods190 T2DM patients were selected as case group and 193 normal persons were select
ed as control group.The genotypic frequencies and the allelic frequencies of rs3731628,rs45788
07, and rs11677500 sites in two groups were compared by  PCR technique and matrix-assisted laser desorption/ionization time of fli
ght mass spectrometry(MAIL-TOF-MS) method; the relationship between the combined effects of multilocus from 3 sites
  of SRC-1  gene and T2DM  was analyzed by haplotype method.Results The genotypic frequencies of rs3731628 and rs11677500 sites of SRC-1  gene did not deviate from Hardy -Weinberg equilibrium in both case and control groups (P> 0.05).The genotypic frequencies of rs4578807 site o f SRC-1  gene deviated from Hardy -Weinberg equilibrium in both case and control groups (P<0.05).There were no significant differences of the frequencies of the genotypes and the alleles of rs37316
28,rs4578807 and rs11677500 sites between two groups(P>0.05).There were no significant differences among the haplotypes formated by every two of rs3731628,r
s4578807 and rs11677500 sites (P>0.05 ).Conclusion There may be no genetic association between the polymorphism of rs3731628 and rs11677500 sites of SRC-1
gene and T2DM in Han population from North China，and SRC-1 gene is not the predisposing gene.


Objective To compare the effects of  immunofluorescence and immunoenzymatic celluar chemistry in detecting the urinary podocytes of I
gA nephropathy (IgAN) patients and to provide  basis for selecting the detection method of  urinary podocytes.
Methods 200 mL morning urine samples were collected from 43 cases who diagnosed as IgAN  (experiment group) and 10 cases of normal control subjects (control
 group) for 3 consecutives days before renal biopsy,and the smears were made from the supernatant by centrifugation.The IgAN patients were classified into Lee Ⅰ
 -Lee Ⅴ groups according to Lee degree method.Immunofluorescence and immunoenzymatic celluar chemistry with monoclonal anti-podocalyxin(PCX) antibody were performed to detect the  urinary podocytes and the number of urinary podocytes was counted  by fluorescence and optical microscope;the correlation between the number of urinary podocytes detected by two kinds of methods were analyzed by  Spearman correlation analysis.
Results There were no urinary podocytes of the objects in   control group detected by  two kinds of  methods,but in
  experiment group they were  visible;the positive rate and the median of  urinary  podocytes detected  by immunofluorescence celluar chemistry were lower than those detected  by immunoenzymatic celluar chemistry  (P<0.05);there was a positive  correlation between two kinds of method
s（r=0.842，P<0.01）.Conclusion Immunoenzymatic celluar chemistry is more effective compared with  immunofluorescence celluar chemistry in detecting the  urinary podocytes.


Objective To explore  the differences of the  expressions of MMP-9 and
 TIMP-1 mRNA in  esophageal squamous cell carcinoma tissue and the matched adjacent normal tissue
 and their relationships. Methods 98 spicemens of surgical removed esophageal carcinoma tissue were chosen for detection,the carcinoma tis
sue and the  adjacent normal tissue  were cut and stored simultaneously from every spicemen,respectively.RT-PCR method was used to detect the expressions of MMP
-9 and TIMP-1 mRNA in  esophageal squamous cell carcinoma tissue and the  adjacent normal tissue;the relationships between the expressions of MMP-9 and
TIMP-1 mRNA in esophageal squamous cell carcinoma tissue and clinicopathological characteristics were analyzed.The relationship between the expressions of MMP
-9 and TIMP-1 mRNA in carcinoma tissue was analyzed by Pearson related analysis method.Results
The positive expression rate of MMP-9 mRNA in carcinoma tissue was higher than that  in    adjacent normal tissue
(χ2=22.873，P=0.000), and the positive expression rate of TIMP-1 mRNA in carcinoma tissue was higher than that
 in   adjacent normal tissue （χ2=7.864，P=0.005）.There was  correlation  between the positive expr
ession of MMP-9 mRNA and the depth of tumor infiltration（χ2=5.022，P=0.025）, but there was no correlation  between the positive expression of MMP-9 mR
NA and lymph node metastasis（χ2=0.617，P=0.432）.There was  correlation  between the positiv
e expression of TIMP-1 mRNA and the depth of tumor infiltration（χ2=4.982，P=0.026）, but there was  no correlation  between the positive expression o
f TIMP-1 mRNA and lymph node metastasis（χ2=0.527，P=0.468）. The  positive expression of MMP-9 mRNA  in
esophageal  carcinoma tissue had positive correlation with the expression of TIMP-1 mRNA（r=2.237，P=0.028）.
Conclusion The expressions of MMP-9 and TIMP-1 mRNA may play a very important role in  the development of esophageal squamous cell carcinoma,however both of them have no affection on lymph node metastasis of esophageal squamous cell carcinoma.


Objective To investigate the expressions of transcription factor Oct-4
 and SOX-2 in cervical cancer and normal cervix  tissue and to explore the possibility of Oct-4 and SOX-2 as the  stem cell markers of cervical cancer.
Methods The expressions of Oct-4 and SOX-2  in 52 cases of cervical cancer tissue and 28 cases of normal cervix tissue
 were detected by immunohistochemical SP   method.The expressions of Oct-4 and SOX-2 in the patients with cervical canc
er with different types were analyzed.Results The expression of Oct-4 protein in cervical cancer tissue(46.1%,24/5
2) was significantly higher than that in normal cervix tissue(7.1%,2/28)(P<0.05);the expression of SOX-2 protein in cervical  cancer tissue(59.6%,31/5
2) was significantly higher than that in normal cervix tissue(10.7%,3/28)(P<0.05).The expressions of Oct-4 and SOX-2 in cervical cancer tissue  were significantly correlated with the histological grade(P<0.05);but not correlated with the age,pathological type,FIGO stage of the patients(P＞0.05).The expressions of OCt-4 and SOX-2 were increased with the decreasing of the differentiation degree,and there were significant differences of the expression frequencies  of OCt-4 and SOX-2 in the patients with different differentiation degrees(P<0.05).The expression of Oct-4 was positively related to the expression of SOX-2 in cervical cancer tissue(P<0.05).
Conclusion The expression frequencies of Oct-4 and SOX-2 in cervical cancer tissue are increased with the decreasing of differentiantion degree of cervical cancer which indicates that Oct-4 and SOX-2 may be used as the markers of cervical cancer stem cells.

Objective To explore the diagnostic values of stroke volume variation (SVV) and central venous pressure (CVP)   in low circulatory
blood volume by comparing  the correlations between SVV,CVP and low circulatory blood volume.Methods 120 patients undergoing partial hepatectomy were randomly selected.After general anesthesia with endotracheal intubation and the begining of the operation, nitroglycerin was administered to the patients via vein  with three separate stages of the dosage regimen of 1,2,and 3 μg?kg-1?min-1.The hemodynamic variables including  stroke volume variation (SVV),central venous pressure (CVP),mean arterial pressure (MAP),heart rate (HR),stroke volume index (SVI),cardiac index (CI) and systemic vascular resistance index (SVRI) of the patients  were detected at 4 time points:before infusion (T0),15 min after infusion of nitroglycerin 1 μg?kg-1?min-1 (T1),15 min after infusion of  2 μg?kg-1?min-1 nitroglycerin(T2),15 min after infusion of  3 μg?kg-1?min-1 nitroglycerin  (T3).The maximum of sensitivity and  specificity and   the receiver operating characteristic curve (AUC) of SVV and CVP  responding to CI were detected byoperating characteristic (ROC) curve; Pearson rank correlation method was used to analyze  the correlation between SVV, CVP and CI.Results The AUC of SVV and CVP responding to CI were AUCCVP 0.713 (95%CI, 0.623-0.804) and AUCSVV 0.930 (95%CI, 0.883-0.977).When the SVV was 12.5%,the sum of sensitivity and specificity was the biggest with  ensitivity of 81% and  specificity of 95%;when the  CVP was 5.5 mmHg,the sum of sensitivity and specificty was the biggest with   sensitivity of 73% and  specificity of 59
%. The value of AUCSVV was larger than that of AUCCVP (P<0.05).The values of Pearson’s correlation coefficient of SVV,CVP and CI were rSVV-CI=－0.671
（95%CI －0.455－－0.67）and rCVP-CI= 0.551（95%CI 0.744－0.591） and there were no significant difference between rSVV-CI  and rCVP-CI (P>0.05).
Conclusion SVV and CVP are good parameters for diagnosis of low circulatory blood volume variation.The predictability of SVV for low circulatory blood volume is higher than CVP,and SVV is more sensitive and specific.

Objective To determine the relationship between body fat percent (BF%) and metabolic syndrome (MS) in healthy examinees with different gender,and to
 clarify that BF% was one of the main risk factors to judge MS.Methods 904 persons (554 male and 350 female) for health examination were recruited，the BF%,body mass index (BMI),waist-to-hip ratio (WHR),systolic blood pressure (SBP),diastolic  blood pressure(DBP),fasting blood glucose (FBG), and blood lipid (BL) were compared between male and female;the correlation between  metabolic indexes and BF% in examinees with different gender 
was detected by Spearman correlation analysis;the correlationship between BF% and the risk of MS in examinees with different gender was analyzed
 by Logistic regression;the optimal cut-point  and reliability of BF% in judging MS were calcuated by  receive operating characteristic (ROC) curve  in  the examinees with different gender.Results The levels of SBP,DBP,FBG,BMI,and WHR in the male examinees  were higher than those in the female(
P<0.05).The levels of BF% in the male examinees were  lower than that in the female(P<0.05).The BF% was positively correlated with the levels of  SBP(r=0.27，r=0.41)，DBP(r=0.27，r=0.34),FBG(r=0.18，r=0.37),total cholesterol (TC)(r=0.19，r=0.31),triglycerides (TG)(r=0.42,r=0.47),
low density lipoprotein cholesterol(LDL-C)(r=0.17，r=0.33) and negatively correlated with the level of high density lipoprotein cholesterol (HDL-
C)(r=－0.28，r=－0.30) in both women and men.After adjustment for all  components of MS, BF% was still the independant risk factor of MS  (OR=1.090,95%CI 1.044-1.137).The risk of MS in the men and women with abnormal increasing of BF% was 1.086 (95%CI 1.030-1.144) and 1.107 (95%CI 1.027-1.192) times the people with normal BF%.In ROC analysis on BF% and MS,the areas under ROC  curve were 0.710(95% CI 0.665-0.754) and 0.811(95%CI 0
.749-0.873) in the male and female,respectively. The optimal cut-points of BF% for MS were 29.050% in the male and 38.550% in the different female.
Conclusion BF% is a risk factor and indicator for MS in the people with  gender.

Objective To explore the condition of chronic diseases and to clarify the risk factors of hypertension,osteoarthrosis,and heart diseases of the elder
ly in Jilin province.Methods A total of 21 435 adult residents aged 18-79 years from 32 cities, counties,and districts of Jilin province were selected by multi
-stage cluster random sampling method in 2012.4 115 residents aged 60-79 years were selected to be the subjects.The  condition of the chronic dis
eases was surveyed by questionnaire survey combined with physical examination method.After being weighted according to complex sampling scheme,the sample was used to describe the prevalence rate of the chronic diseases.Multiple Logistic regression analysis was used to analyze the risk factors of the chronic diseases.Results The prevalence rate of chronic diseases among the elderly in Jilin province was 79.5% in the past 1 year.The
 elderly  suffered from one kind of  chronic disease accounted for 22.7%,and the elderly suffered from two kinds of   chronic diseases accounted for 19.8%,and those who suffered from three kinds of  or more chronic diseases accounted for 37.0%.The top ten chronic diseases among the elderly were hy
pertension(33.2%),osteoarthrosis(31.9%),heart diseases(25.1%),digestive system diseases(24.2%),cerebrovascular diseases(16.3%),diabetes mellitus(13.8%),respiratory system diseases(10.6%),diseases of eye and adnexa (9.0%),rheumatism(4.0%),and hyperlipidemia(2.8%).The results of analysis on health-related behavior indicated that there were significant differences between different gender in smoking,drinking,taste,physical exercise
,sleep and body mass index(BMI),respectively(P<0.05).The Multiple Logistic regression analysis results indicated that gender,exercise, and BMI were main influencing factors of hypertension;gender and income were influencing factors of osteoarthrosis;gender,age,occupation,drink,and BMI were influencing factors of heart diseases.Conclusion Compared with other provinces and areas,the prevalence  rate of chronic diseases in Jilin province among the elderly is high.Hypertension,osteoarthrosis,and heart diseases are the top three kinds of diseases among all chronic diseases.The influencing fa
ctors of hypertension,osteoarthrosis and heart diseases are gender,age,occupation,income,drinking,exercise,and BMI.

Objective To describe the general mental health status and its distribution characteristics among the adults in Jilin province,and to investigate th
e influencing factors of mental health.Methods In July to August of 2012,the adults  in  Jilin province were surveyed with 12-item general health questionnaire (GHQ-12) by the method of multi-stage and stratified random cluster sampling;the influencing factors of mental health status were analyzed by Logistic regression analysis. Results After being weighted,the average score of GHQ-12 was 2.27±2.11 and the  positive rate of GHQ-12 was 23.8% among the adults in Jilin province. The GHQ-12 positive rate of the female was higher than that of the male (27.9%  vs  20.0 %,P<0.001). The adults
 in rural area had a higher GHQ-12 positive rate compared with the  adults in urban areas (25.9% vs 22.2%,P<0.001).The Logistic regression analysis results showed that the risk factors of mental health were  female,marriage situation as unmarried,divorced and widowed,occupation as farmers,students and unemployment,the average daily sleep time less than 7 h,and having chronic diseases within one year;and the age over 35 years old,education as junior middle school,undergraduate and above,average income per family over 500 yuan monthly,regular dietary,and physical exercises were the protective factors of mental health.Conclusion The GHQ-12 positive rate of general mental health among the adults in Jilin province is higher than those in most of the rovinces in China.Besides,the mental health status of populations of female,farmers,students,unoccupied people,lower education,unmarried or poor marriage eople,lower income,and bad conditions in the dietary,sleep and body should be paid more attention　to.

Objective To investigate the situation of smoking,passive smoking and quitting smoking,and the cognitive and the attitudes towards tobacco-rela
ted harm among adults in Jilin province,and to provide basis for formulatting tobacco control policies in Jilin province.Methods By multi-stage stratified ran
dom cluster sampling methods,a total of 21 435 adults aged 18-79 years old  were enrolled in the questionnaire survey and physical examination.After being weighted according to the complex sampling scheme and post-stratification,the sample was used to estimate the prevalence rates of smoking,quitting smoking and passive smoking,and the cognitive of tobacco-related knowledge and attitudes towards smoking control of adults in Jil
in province.Results The prevalence  rate of smoking was 31.8%,and it was higher  in the  male than the female (52.9%  vs  9.4%，P<0.001),and it was
higher in rural area than  urban area (33.2%  vs  30.7%，P<0.01). The prevalence rates of smoking were significantly different among the adults with different ages,educations,marriages,occupations and month  incomes per family (P<0.001).The prevalence rate of quitting smoking of the adults in Jilin province was 18.6%,and the rates  were significantly  different among the adults with different ages,marriages,occupations and month incomes per family (P<0.001).The passive smoking rate of the adults in Jilin province was 60.5%.People encountered passive smoking mostly at home and  workplace (33.4% and 21.7%).The percentages of supportting non-smoking in public places and restaurants and increasing the efforts of government’s tobacco contr
ol  were 93.4%,92.0%,and 93.9%,respectively.The cognitive rate of tobacco-related knowledge of the female was higher than the  male (P<0.001),and the
cognitive  rate of the people in rural was less than that in  cities (P<0.05).The cognitive rate of “the harm of low-tar content cigarettes being similar to common cigarettes” was low,which was only 15.3%.ConclusionThe prevalence rates  of smoking and passive smoking are higher than those in 20
02, and the cognitive  rate  of tobacco-related knowledge is low,and the rate of supporting tabocco control  of the  adults in Jilin province is high.