Objective:To observe changes in the morphology and activity of astrocytes in spinal cord at different temperatures after scratches treatment, and to investigate the effect of mild hypothermia on hyperplasia of reactive astrocytes. Methods:Spinal cord astrocytes were cultured from the neonatal SD rat, and reactive astrocytes were prepared by scratches treatment. Mild hypothermia choose 33℃, cell cul-ture 48 h. Cells were divided into control group、scratches group、mild hypothermia group and scratch+mild hypothermia group. Cell mor-phology of each group were observed in 0 h、6 h、12 h、24 h、48 h、3 d、5 d、7 d. Nestin positive rate were detected by using immunofluorescence staining method. Cell activity were observed by methyl thiazolyl tetrazolium salt (MTT) assay. The degree of apoptosis was observed by using PI staining method. Results:Compared with control group and mild hypothermia group, cell body in both scratches group and scratch+mild hypothermia group was hypertrophy, the protrusion around was increased and extend, the cytoplasm was enrich, and the growth rate was signif-icantly increased. Compared with scratches group, scratch+mild hypothermia group cells grew slowly, the protrusion around was decreased, and cells of the scratched area ingrowth significantly slowly. Nestin and PI positive rate of cells were also significantly lower. All the results had statistically significant differences (P<0.01). Conclusion:After scratch injury, astrocytes were activated to be reactive astrocytes and hyper-plasia. Mild hypothermia significantly inhibited spinal cord reactive astrocytes overgrowth and restrain astrocytes apoptosis.

Objective:To investigate the effects ofβ-sheet breaker peptide H102 on expression of synaptic plasticity associated proteins and learning and memory functions in double transgenic Alzheimer's disease(AD) mice,and to discuss its mechanisms. Methods:Thirty APP-swe/PS1dE9 double transgenic male mice of 8 weeks were randomly divided into model group and H102 treatment group (15 mice per group). In addition,a group of C57BL/6J mice with the same age and background was set as normal. H102 (5.8 mg/kg) 5 μl was infused by in-tranasal administration to mice in H102 treatment group,and equal volume of blank solution of H102 (chitosan,BSA) was given to mice in con-trol group and model group. The ability of spatial reference memory was tested by Morris Water Maze after 16 weeks treatment,then immunohis-tochemistry tests and Western blot technique were used to detect the content ofβ-amyloid peptide(Aβ_1-42) protein and phospho protein kinase C α、β₂、γ(p-PKCα, p-PKCβ₂, p-PKCγ), phospho-N-methyl-D-aspartate receptor1(p-NMDAR1), phospho-Calcium/Calmodulin dependent pro-tein kinaseⅡα(p-CaMKⅡα) and phospho-cAMP response element binding protein(p-CREB) of synaptic plasticity associated proteins in mice brain. Results:The ability of learning and memory was significantly improved in H102 treatment group than that in model group by the test of Morris Water Maze. The contents of Aβ_1-42 proteins and p-PKCα, p-PKCβ₂, p-PKCγ, p-NMDAR1, p-CaMKⅡαand p-CREB of synaptic plas-ticity associated proteins in mice brain were improved significantly in H102 treatment group than those in model group by the test of immunohis-tochemistry tests and Western blot technique. Conclusion:β-sheet breaker peptide H102 can significantly improve synaptic plasticity and the ability of learning and memory in double transgenic AD mice.

Objective:To investigate the effects of prenatal stress on astrocytes after ischemia/reperfusion of cerebral middle artery in adult offspring rats. Methods:Pregnant rats were randomly assigned to prenatal stress treatment group, which was exposed to restraint three times daily in the last week of pregnancy, and no prenatal stress treatment group. Adult male offspring rats were subjected to transient focal cerebral ischemia by middle cerebral artery occlusion (MCAO). There were three groups:prenatal stress+sham group, MCAO group and prenatal stress+MCAO group (n=10). After 5 days of reperfusion, the infarct size was evaluated. The morphology of astrocytes, co-local-ization of erythropoietin-producing hepatocellular receptor A4 (EphA4) and glial fibrillary acidic protein (GFAP) were detected by double im-munofluorescent staining. And the protein expressions of EphA4, GFAP and Neurocan in peri-ischemic regions were detected by Western blot. Results:The infarct size and the expression of EphA4, GFAP and Neurocan were significantly increased in prenatal stress+MCAO group compared with MCAO group (all P<0.05). And the morphological changes of GFAP-positive astrocytes and co-localization of EphA4/GFAP were more obvious in prenatal stress+MCAO group compared with MCAO group. Conclusion:Prenatal stress may upregulate the expression of EphA4 on astrocytes in the offspring rats after cerebral ischemia/reperfusion, which promotes the reactivity of astrocyte and increases the ex-pression of neurocan.

Objective:To study the effect of leptin on neuron apoptosis in mice with cerebral ischemia injury. Methods:Seventy-five male Kuming mice were randomly divided into 3 groups:sham, model and leptin intervention group, respectively. Focal cerebral ischemia/reperfusion injury model in mice was established by middle cerebral artery occlusion. Leptin intervention group was injected with leptin (1μg/g weight, I. P.) at 0 min of ischemic injury. Neuron apoptosis was detected by TUNEL staining. The mRNA expression of apoptosis relative gene bcl-2 and caspase-3 were detected by RT-PCR. The protein expression of bcl-2 and caspase-3 were detected by immunohistochemistry. Results:In model group, most of the neurons in the central area of cerebral ischemia had necrosis obviously, and the amount of neuron apop-tosis was much higher than that in sham group (P<0.01). Compared with sham group, both expression of pro-apoptosis gene caspase-3 and anti-apoptosis gene bcl-2 increased significantly in model group (P<0.01). Compared with model group, the amount of neuron apoptosis and expression level of caspase-3 were decreased significantly (P<0.01), whereas the mRNA and protein expression of bcl-2 were increased sig-nificantly in leptin intervention group (P<0.01). Conclusion:Leptin could reduce neuron apoptosis through down-regulation the expression of caspase-3 and up-regulation the expression of bcl-2. It suggests that leptin could play a neuroprotective role in cerebral ischemia injury.

Objective:To investigate the protective effects of endogenous gangliosides on LPS-induced PC12 cells injury and its possible mechanism. Methods:PC12 cells were cultured, and lipopolysaccharide(LPS) injury model was established. We detected the changes in sur-vival rate of different concentrations of LPS on PC12 cells, and the changes in survival rate of LPS when endogenous gangliosides were exhaust-ed by D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol(D-PDMP), and the protective effects of monosialoganglioside (GM1) on LPS-induced PC12 cells injury. Meanwhile, we observed the morphological changes of GM1 on PC12 cells induced after LPS injury by inverted microscope and fluorescence microscope, and then we detected the relative expression of NF-κB. Results:LPS could decrease the survival rate of cells in a concentration-dependent manner, and GM1 could significantly protect the cells against the changes in survival rate and morphologi-cal damages induced by LPS; After depletion of endogenous gangliosides by D-PDMP, LPS had more injury to PC12 cells, which could be im-proved by adding GM1; RT-PCR results showed that the relative expression of NF-κB in PC12 cells was increased in LPS group, while relative expression of NF-κB increased much higher when endogenous gangliosides were exhausted by D-PDMP, and the relative expression of NF-κB was decreased after GM1 being added. Conclusion:The endogenous gangliosides might through NF-κB signal pathway to protect PC12 cells a-gainst LPS-induced injury.

Objective:To research the effects of gene polymorphisms of AGT at G-217A and T174M loci on incidence and the hypoxia acclimation to acute mountain sickness (AMS). Methods:Step 1:61 Han students origin low landers were exposed in acute hypoxia at 4 800 m altitude-equivalent for 6 hours, and took the supine bicycle exercise for 20 min at quantitative load of 60 r/min, 80 W after entering the cabin for 30 min, then the AMS were evaluated by the Lake Louise acute mountain sickness scoring system (LLS). The physical index of heart rate (HR), ambulatory blood pressure and oxygen saturation (SpO₂) were measured during exercise. Step 2:The physical index was measured again in acute hypoxic exposure after 3 weeks increasing hypoxia exercise (At an increasing altitude of 2 500 m, 3 500 m, 4 800 m, time of 2 hours/day, 4 days/week, and in moderate intensity of exercise). PCR-RFLP was used to determine the genotypes and alleles frequencies of AGT at G-217A and T174M loci. Results:In step 1 the hypoxic exposure, for the G-217A locus in AGT, there was no significant difference between GG and GA+AA gene tester. While in step 2 the hypoxic exposure, the SpO₂ of GG gene was lower than GA+AA gene obviously (P<0.05). No significant differences of AMS incidence, VE, SpO₂, HR and blood pressure were detected in different genotypes and alleles at T174M locusin both hypoxic exposures. Conclusion:The G-217A may be the genetics sign of hypoxia acclimation. There is no obvious rel-evancy between the T174M polymorphism and the occurrence of AMS/hypoxia acclimation.

Objective:To investigate the protection effects of hypoxic preconditioning(HPC) on the SH-SY5Y cell injured by oxygen-glu-cose deprivation(OGD),and to discuss the possible mechanism. Methods:SH-SY5Y cells were randomly divided into 4 groups. In normal group,the cells were cultured without OGD treatment. In HPC group,the cultured SH-SY5Y cells were treated for 5 days by intermittently ex-posing to hypoxic gas mixture (2% O₂,5% CO₂) for 30 min in every day. In OGD group,the culture medium was replaced by glucose-free medium and the cells were transferred to a humidified incubation chamber flushed by a gas mixture of 1% O₂ and 5% CO₂ for 10 h. After that, the cells were fed with glucose-supplemented medium and cultured under normoxic condition for 24 h. In HPC+OGD group,the cultured SH-SY5Y cells were treated for 5 days by intermittently exposing to hypoxic gas mixture for 30 min in each day, then the cells were given the same treatments as those in OGD group. The cell viability was assessed by MTT assay. The degree of the cell damage was evaluated by deter-mining lactate dehydrogenase (LDH) leakage. TUNEL staining were used to detect the variation of cell apoptosis. The expression of Caspase 3 and hypoxia inducible factor-1α(HIF-1α) at protein levels was examined by Western blot. Results:Hypoxic preconditioning relieved the cells apoptosis,decreased the amount of LDH leakage and improved the viability of SH-SY5Y cells injured by OGD (P<0.05). Western blot showed that the expression of Caspase 3 protein in HPC+OGD group was significantly lower than that in OGD group (P<0.05); HIF-1α protein expression was significantly higher than that of OGD group (P<0.05). Conclusion:Hypoxic preconditioning has protective effect on in vitro cultured SH-SY5Y cells injured by OGD. The mechanism may be related to the increase of HIF-1a protein.

Objective:To investigate the effects of supplement of corn peptides plus aerobic exercise on fat loss and blood lipid profile in obese rats resulted from high fat diet and the mechanism:the role of adipose triglyceride lipase (ATGL) and lipoprotein lipase (LPL). Methods:One hundred and fifty male SD rats (4 weeks age) were randomly divided into control group (C, n=15) fed with ordinary diet and obese model group (M, n=135) fed with high fat diet for 8 weeks. Forty obese rats whose body weight increased by 20% of the mean value of con-trol group were selected and randomly divided into 5 groups(n=8):obesity control group, casein group, corn peptides group, exercise group and exercise+corn peptides group. The rats of the latter two groups completed aerobic excise for 4 weeks at speed of 15 m/min and duration of 60 min per time (6 times/week). After 4 weeks of intervention, the bloods of rats were collected and blood lipid profile plasma levels of triglyceride(TG), total cholesterol (TC), high density lipoprotein(HDL), low density lipoprotein(LDL) were detected. The perirenal fat and epididymal fat were collected and weighed, and the protein levels of ATGL in livers and LPL in adipose tissues were detected by Western blot. Results:①Compared with obesity control group, the body weight and the masses of perirenal fat and epididymal fat in exercise group and corn peptides+exercise group were decreased obviously, with more obvious decrease in the corn peptides+exercise group compared with the exer-cise group. No difference was found in the rats of corn peptides group and casein group. ②Compared with obesity control group, the plasma TG was decreased in exercise group, and the plasma levels of TG and TC were reduced in the rats of corn peptides+exercise group, while they remained unchanged in the rats of corn peptides group and casein group. The plasma levels of HDL and LDL had no significant difference a-mong groups. ③The protein levels of ATGL in livers and LPL in adipose tissues were significantly increased in the rats of exercise group and corn peptides+exercise group, with more obvious in the latter group. No difference was shown in the rats of corn peptides group and casein group compared with obesity control group. Conclusion:Significant decrease of body weight, perirenal fat and epididymal fat as well as plasma TG, TC were induced by 4-week aerobic exercise or supplement of corn peptides plus exercise, with more obvious effects induced by corn pep-tides plus exercise than exercise, while no effect was induced by corn peptides alone, which may be related to the enhancements of the protein levels of ATGL in livers and LPL in adipose tissues.

Objective:To investigate the role of sub-transform macrophage in ischemia/reperfusion renal injury in rats, as well as under-lying mechanisms. Methods:Thirty male Sprague-Dawley rats were randomly divided into ischemia/reperfusion (IRI, n=24, renal artery was occluded for 45 min) group and sham-operation (Sham, n=6) group. The kidneys in IRI group were collected at 0, 6, 24 and 72 h after operation (6 rats for each time point). The injury of the kidney was detected with HE staining. Immunohistochemistry staining was performed to evaluate the expression of proliferating cell nuclear antigen (PCNA). Real-time PCR was used to detect the mRNA expression of macrophage migration inhibitory factor (MIF). Moreover, the expression and location of MIF, monocyte chemoattractant protein-1 (MCP-1) and macrophage marker CD68 were examined by immunofluorescence staining. Most importantly, the distribution of macrophage subtypes M1 and M2 was analyzed by flow cytometry. Results:The worst pathologic damage of the renal tissues, as well as infiltration of inflammatory cells, was observed at 24 h after operation in IRI rats, with obvious recovery afterwards. Immunohistochemistry staining showed that the expression of PCNA was significantly increased after the ischemia/reperfusion, peaking at 6 h and reducing at 72 h after operation. Compared with sham group, the levels of MIF at mRNA and protein levels were both significantly increased after the ischemia/reperfusion, while the expression of MCP-1 was peaked at 6 h and decreased afterwards. Moreover, the expression of CD68-positive macrophages were significantly increased in IRI rats, with peaking at 24 h and reducing at 72 h. Furthermore, after 6 h of reperfusion, the percentage of M1 macrophages reached the peak, and thereafter the relative expression of M1 and M2 was reduced and increased, respectively. Conclusion:In the early phase of ischemia/per-fu sion renal injury, M1 macrophage results in renal damage, and afterwards the M2 macrophage is increased and repairs the renal damage by improving the cell proliferation.

Objective:To establish an easy, not depending on advanced laboratory apparatus method to isolate and culture rat pulmonary artery smooth muscle cells (PASMCs), and to explore the effects of platelet-derived growth factor (PDGF) on cell proliferation and migration. Methods:The right ventricle was perfused with the mixture of iron, agarose, and the PASMCs and iron could adhere to agarose. The iron-con-taining tissue would move to side of the tube next to the magnet and could be digested by collagenase I. By the method, vessel-containing tissue could be attained. With 3-4 weeks' purification, the PASMCs could be obtained. The PASMCs morphology was observed by an inverted micro-scope, and identified by immunocytochemistry and immunofluorescence. The effects of PDGF on cell proliferation and migration was detected by MTT assay and scratch wound assay. Results:14 days、21 days and primary culture after isolation, the PASMCs was identified, and the re-sult showed that isolation and primary culture of the cells were PASMCs. Compared with the cells with no stimulation, the proliferation of PASMCs exposed to PDGF was increased significantly(P<0.05), and scratch wound assay demonstrated that PDGF induced the significant increase of migration of PASMCs. Conclusion:This method to isolate and culture rat PASMCs is simple, not depending on advanced laborato-ry. PDGF can promote the proliferation and migration of PASMCs.

Objective:To observe the effects of low-concentrations of alcohol consumption on the expression of mitofusin-2 (mfn2) in myocardial injury of diabetic rats. Methods:Diabetic rat model was simulated by intraperitoneal injection of 55 mg/kg streptozotocin (STZ) and divided into control group, diabetes mellitus(DM) and diabetes+ethanol (DM+EtOH) groups (n=6). When diabetic model was suc-ceed, daily consumption of 2.5% ethanol was used in ethanol+diabetic group after one week, then changed to 5% ethanol continued until 8 weeks. Eight weeks after the modeling, heart perfusion ex vivo. The ventricular hemodynamic parameters were recorded, the serum levels of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) were determined by automatic biochemistry analyzer, the mfn2 protein ex-pression of left anterior myocardium was evaluated by Western blot and immunohistochemistry. Results:Compared with control group, the left ventricular development pressure (LVDP), heart rate (HR) and rate pressure product (RPP) were decreased, however, left ventricular and diastolic pressure (LVEDP) and LDH, AST release were increased, the expression of mfn2 protein was decreased in DM group. Compared with DM group, LVDP, HR and RPP and the expression of mfn2 protein were increased, LVEDP and LDH, AST were decreased in DM+E-tOH group. Conclusion:The expression of mfn2 protein was decreased in myocardial injury of diabetic rats, low-concentrations of alcohol consumption increased the expression of mfn2. It suggests that mfn2 may be participated in the cardiac protective role of low-concentrations al-cohol intervene in diabetic rat.

Objective:To study the role of Notch1 signaling pathway in the myocardial protective effects of resveratrol pro-treatment after hepatic ischemia/reperfusion injury. Methods:Thirty-six healthy male SD rats were randomly divided into 3 groups:sham control (SC) group, hepatic ischemic/reperfusion (HIR) group, and pro-treatment resveratrol (Res) group,12 rats in each group. After each group with hepatic ischemia 40 min reperfusion 2 h (SC group placing equal time), the left ventricular function including left ventricular pressure (LVP), left ventricular systolic pressure (LVSP) and its derivate (±dp/dt) was measured; the serum activities of creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) and tumor necrosis factorα(TNF-α) and interleukin-6(IL-6) were detected. The activity of myocardial su-peroxide dismutase(SOD) and contents of myocardial malonaldehyde (MDA) were determined; Notch intracellular domain (NICD) expressions were detected with Western blot; the mRNA levels of Notch1 and TNF-αwere detected by real-time PCR. Results:Compared with SC group, the left ventricular functions in the HIR group were significantly decreased. With decreasing the expression of NICD and Notchl, the activities of CK-MB, LDH, TNF-αand IL-6 in the serum were evidently increased(P<0.01), the activity of SOD in myocardum was decreased and contents of MDA in myocardum was enhanced (P<0.01). Compared with HIR group, the left ventricular functions were significantly en-hanced in Res group (P<0.01). The above mentioned parameters in the serum in the Res group were evidently improved (P<0.01), meanwhile the expression of NICD and Notchl were significantly enhanced whereas the expression of TNF-α was obviously decreased. Conclusion:The findings indicate that resveratrol has protective effect on myocardial injury during the hepatic ischemia/reperfusion, and its mecha-nisms may be related to anti-inflammation and anti-oxidative stress via Notch signaling pathway.

Objective:To investigate the effect of Dexmedetomidine (Dex) on Toll-like receptor 4(TLR4) expression in lung during lung ischemia/reperfusion(I/R) in rats and its possible protecting mechanisms. Methods:In vivo I/R model in left lung of SD rats was estab-lished. Fifty adult healthy male SD rats were randomly divided into five groups (n=10):control group (Sham group), I/R group, Dex group, atipamezole group (Atip group) and Dex+Atip group. After the I/R experiment,rats were killed and the left lung tissues were harvest-ed to get the lung wet/dry weight(W/D); Ultrastructure of lung tissue were observed under light microscopy; The mRNA expression of TLR4 in lung tissues were determined by RT-PCR; The protein level of TLR4 in lung tissues was detected by Western blot. Results:①Compared with those in the Sham group, W/D and total lung water content (TLW) in other groups increased significantly (P<0.05), the mRNA and protein expression levels of TLR4 in lung tissues increased too. The structure damages of lung tissues observed under light microscopy in other groups were more than that of Sham group. ②Compared with those in the I/R group, W/D and TLW in the Dex group were lower (P<0.05, P<0.01), the mRNA and protein expression levels of TLR4 in lung tissues decreased (P<0.01), and reduced structure damages of lung tissues were observed under light microscopy in Dex group. ③Compared with those in the Dex group, W/D and TLW in the Dex+Atip group were higher (P<0.01), the mRNA and protein expression levels of TLR4 in lung tissues increased (P<0.01), and the structure damages of lung tissues observed under light microscopy were more serious. There was no significant difference of the above parameters among I/R、Atip、Dex+Atip groups. Conclusion:Lung ischemia/reperfusion caused high expression of TLR4 and finally induced damages of the lung. Dexmedetomidine could inhibit TLR4 expression and alleviate the lung ischemia/reperfusion injury, which was related to activation of α2-adreno receptor.

Objective:To study the effects of Ligustrazine on the Ca²⁺ sensitivity in carotid artery smooth muscle of simulated weightless-ness rats. Methods:Twenty-one female SD rats were divided into control group (CON), simulated weightlessness group (TS), simulated weightlessness plus Ligustrazine administration group (LTZ) (n=7). After 4 weeks, the contractile property and the Ca²⁺ sensitivity in carotid artery were measured. Furthermore, the effects of ML-7(a myosin light chain kinase specific inhibitor) on above mentioned parameters were recorded. Results:Compared with CON group, the contractile force induced by phenylephrine (PHE) in TS group was increased by 25.14% (P<0.01). The force in LTZ group decreased by 13.46% (P<0.01) and increased by 8.30% compared with TS and CON group, respec-tively. After the artery rings were incubated with ML-7, the force in CON, TS and LTZ group were reduced by 8.84%, 16.24% (P<0.01) and 3.40%, respectively. Compared with CON group, the contractile force induced by KCl in TS group was increased by 40.46% (P<0.01). The force in LTZ group decreased by 18.80% and increased by 14.05% compared with TS and CON group, respectively. The force in CON, TS and LTZ group were reduced by 21.97% (P<0.05), 21.88% (P<0.01) and 10.84% by ML-7, respectively. Compared with CON group, the pD2[Ca²⁺] in TS group was increased by 10.03% (P<0.01). The pD2[Ca²⁺] in LTZ group was decreased by 7.01% (P<0.01) and increased by 2.32% compared with TS and CON group, respectively. The pD2[Ca²⁺] in CON, TS and LTZ group was reduced by 2.42%, 7.43% (P<0.01) and 2.51% by ML-7, respectively. Conclusion:The contraction of carotid artery is strength-ened in simulated weightlessness rats, it maybe results from the Ca²⁺ sensitivity of contractile proteins in smooth muscle increasing. Ligus-trazine can improve the contraction of carotid artery, reducing the Ca²⁺ sensitivity and inhibiting myosin light chain kinase may be involved in its mechanisms.

Objective:To observe the expression of 5-Hydroxytryptamine 2B receptor (5-HTR2B)、E-cadherin (E-cad)、alpha-smooth muscle actin(α-SMA) in the bleomycin -induced pulmonary fibrosis tissue of rats. Methods:Forty-five healthy male SD rats were randomly di-vided into control group,bleomycin group and bleomycin+prednisone group(n=15). Five rats in each group randomly sacrificed at the 7^th、the 14^th and the 28^th day after eastablishing models. The lung tissue was observed by microscope in HE and Masson staining. Lung hydroxypro-line (HYP) content was evaluated. The expression of protein and mRNA of 5-HTR2B, E-cad and α-SMA were analyzed by immunohistoche-mistry and/or RT-PCR. Results:A dynamic changes from alveolitis to pulmonary fibrosis could be observed in the slices by HE and Masson staining. The experimental results by immunohistochemistry and RT-PCR showed that the protein and mRNA expression of 5-HTR2B and a-SMA enhanced rat pulmonary fibrosis (P<0.05) and reached the highest at the 28^th day; the corresponding protein and mRNA expression of E-cad reduced (P<0.05), and reduced to the lowest value at the 28^th day. Compared with bleomycin group, the corresponding mRNA and protein expression of 5-HTR2B and a-SMA in bleomycin+prednisone group had decreased (P<0.05); however, the corresponding mRNA and protein expression of E-cad had increased(P<0.05). Conclusion:5-HTR2B is involved in the pathogenesis of pulmonary fibrosis throught epithelial-mesenchymal transition (EMT).

Objective:To observe the effects of arecoline on proliferation and apoptosis of MCF-7 human breast cancer cells and to ex-plore its possible mechanism. Methods:Human breast cancer MCF-7 cells were treated with arecoline at the concentrations of 0,10,30,50, 100,300,500μmol/L, the cell proliferation were detected by MTT assay, cell apoptosis were analyzed by Hoechst 33342 staining and flow cy-tometry, the protein expression of Bax,Bcl-2 and P53 were detected by Western blot. Results:Low concentration(0,10,30, 50 μmol/L) arecoline had no effect on the proliferation and apoptosis of MCF-7. However, high concentration(100,300,500μmol/L) arecoline inhibited proliferation and induced apoptosis of MCF-7 cells in a concentration-dependent manner, arecoline also significantly increased P53 and Bax protein expression and decreased Bcl-2 protein expression. Conclusion:High concentration arecoline inhibited the proliferation and induced the apoptosis of MCF-7 cells, the mechanism was probably corrected with increasing P53 and Bax protein expression and decreasing Bcl-2 pro-tein expression.

Objective:To study the protective effects of Trillium tschonoskii maxim (TTM) on rats' oxidative stress and hepatotoxicity in-duced by lipopolysaccharide (LPS). Methods:Sixty SD rats were randomly divided into TTM high, medium and low dose groups, model group, Dexamethasone (DEX) control group and blank control group with ten rats in each group. The TTM high, medium and low dose groups were treated with 8, 4, 2 g/(kg·d) TTM by intragastric administration and model group, DEX control group and blank control group were treated with the same amount of distilled water respectively. The TTM high, medium and low dose groups, model group, DEX control group were injected intraperitoneal with 1 mg/kg LPS and the DEX control group was injected intraperitoneal with 5 mg/kg DEX, the blank control group was injected with same amount of normal saline every five days. The indexes of rats' thymus and spleen were measured in 30 days. The activities of serum nitric oxide synthase (NOS), superoxide dismutase (SOD) and the contents of nitric oxide (NO), glutathione (GSH), glu-cosinolates barbituric acid reaction product(TBARS), white cells interleukin-6(IL-6), IL-10 and tumor necrosis factor-α(TNF-α), the activi-ties of liver SOD, GSH-Px and the contents of GSH and TBARS were measured. Results:TTM high dose group was significantly different in body weight in 19~30 days(P<0.05); The index of thymus in TTM high, medium and low dose groups and the index of spleen in TTM high dose group were decreased significantly compared with those of the model group. The activity of serum NOS and the contents of TBARS and NO in TTM high, medium and low dose groups were decreased significantly(P<0.05). The activity of serum SOD in TTM high dose group and the contents of GSH in TTM medium and low dose groups were increased significantly(P<0.05). The contents of serum IL-6 and TNF-αin TTM high, medium dose groups were decreased significantly and the contents of serum IL-10 were increased significantly(P<0.05). The con-tents of liver TBARS in TTM high, medium dose groups were decreased significantly. The activity of liver SOD in TTM high, medium and low dose groups, the activity of GSH-Px in TTM high, medium dose groups and the contents of GSH in TTM high dose group were increased signifi-cantly(P<0.05). Conclusion:TTM has a certain effect to delay the rats' atrophy of thymus and spleen generated by LPS. It can effectively reduce the activity of NOS in serum, reduce the formation of NO, improve the activity of SOD, GSH-Px and the contents of GSH, reduce lipid peroxidation, decrease the excessive secretion of IL-6, TNF-α, increase the contents of IL-10, which can resist inflammation and protect the liver.

Objective:To investigate the effects of pirfenidone on CCl4-induced liver fibrosis in mice. Methods:After 8-week feeding, 40 healthy male SPF ICR mice were randomly divided into 4 groups:liver fibrosis group (CCL₄ group), low doses of Pirfenidone group (PFD-L group), high doses of Pirfenidone group (PFD-H group) and control group. The mice in CCL₄ group, low doses of Pirfenidone group (PFD-L group), high doses of Pirfenidone group (PFD-H group) were injected intraperitoneally with 0.4 ml 10% CCL₄ solution dissolved in soybean oil. Then the PFD-L and PFD-H groups were treated with 120 mg and 240 mg PFD via gastric gavage, respectively. Control group was injected with same volume of saline. Alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) in serum were tested with automatic biochemistry analyzer and the pathologic changes of liver tissue were examined by HE staining. Furthermore, we identi-fied hyaluronic acids(HA), laminin(LN), collagentype IV(IV-C) in serum using radioimmunoassay and the expression of smooth muscle acti-nalpha(α-SMA) related gene in liver was tested by real-time fluorescence quantitative PCR. Results:Compared with control group, hepatic lobules in CCL₄ mice were damaged significantly, collagenous fiber was deposited obviously, and counterfeit hepatic lobules formed. The serum levels of ALT, AST, ALP were increased obviously (P<0.05) with the enhancement of HA, LN, IV-C in serum (P<0.05) and the ex-pression of α-SMA related gene (P<0.05). Compared to CCL₄-treated mice, the serum levels of ALT, AST, ALP in PFD-L and PFD-H groups were decreased, HA, LN, IV-C in PFD-L and PFD-H mice went down obviously,and the expression of α-SMA related gene was con-trolled (P<0.05). From pathological observation, we found the degree of liver fibrosis in PFD-L mice was reduced and collagenous fiber was decreased, only a little counterfeit hepatic lobule could be found. Cell arrangement in PFD-H mice recovered, the structural of hepatic lobules disordered and no obvious counterfeit hepatic lobules were found. Therefore, the recovery of PFD-H group was better than PFD-L group.Conclusion:Pirfenidone has a protective role in improving the outcome of the liver fibrosis and it may become a new direction of early intervention in liver fibrosis.