Objective To investigate the effects of ulinastatin on the expression of NF-κB in lung tissues during limb ischemia-reperfusion(I/R) in rats.Methods Forty-eight male SD rats weighing 230-260 g were randomly divided into 3 groups (n=16 each):sham operation group (group S);I/R group; ulinastatin group (group U).A rat model of lung injury induced by I/R of hind limbs which was produced by occlusion of the bilateral femoral arteries for 2 h followed by reperfusion was established.The rats were sacrificed at 2 and 4 h of reperfusion(8 rats at each time point) and the lung tissues removed for determination of NF-κB expression (by immuno-histochemistry) and microscopic examination.W/D lung weight ratio was calculated.Results W/D lung weight ratio was significantly increased and NF-κB expression was up-regulated in group I/R compared with group S(P＜ 0.05). W/D lung weight ratio was significantly decreased and NF-κB expression was down-regulated in group U compared with group I/R (P＜0.05). The lung injury induced by I/R was reduced in group U compared with group I/R. Conclusion Ulinastatin can reduce the lung injury induced by limb I/R by down-regulating the expression of NF-κB in rat lung tissues.

Objective To evaluate the effects of propofol on apoptosis and invasiveness of human lung cancer cell line A549 cells.Methods Human lung cancer cell line A549 were seeded onto 96-well plates (100 μl/well) and 6-well plates (2 000 μl/well) at a density of 2× 105 cells/ml,and cultured for24 h at 37 ℃ in 5％ CO2.The cells were randomly divided into 2 groups (n =60 each) using a random number table:dimethyl sulfoxide (DMSO) group and propofol group (group P).In group P,propofol with the final concentration of 100 μmoYL was added.In group DMSO,0.5％ DMSO with the final concentration of 0.5％ was added.At 24 h of incubation with drugs,caspase-3 expression was detected by high content screening (HCS); the expression of matrix metalloproteinase (MMP-2) was detected by Western blot analysis.At 0.5,1 and 5 h of incubation,ERK1/2 expression was also measured using Western blot analysis.Results Compared with group DMSO,the expression of caspase-3 was up-regulated,the expression of MMP-2 was down-regulated,ERK1/2 expression was up-regulated at 0.5 of incubation and down-regulated at 1 h of incubation,and no significant change was found in ERK1/2 expression at 5 h of incubation in group P.Conclusion Propofol can promote apoptosis in A549 cells and inhibit invasiveness of human lung cancer cell line A549 cells.

Objective To investigate the effects of anaesthetic concentration of sevoflurane on TM3 mouse leydig cell viability.Methods TM3 mouse leydig cells were randomly divided into 3 groups ( n =24 dishes each):control group (group C),2％ and 5％ sevoflurane groups (groups SEV1 and SEV2 ).The cells were collected after being exposed to sevoflurane or 95 ％ room air + 5 ％ CO2 for 2,4 or 6 h (T1-3) for microscopic examination with optical binocular inverted microscope.The number of live cells was counted by using cell counting kit8.Gene chips were used to indentify differentially expressed genes between group C and group SEV2 after being exposed to air and 5 ％ sevoflurane for 6 h respectively.Results The leydig cell viability was significantly decreased at T3 in group SEV2 as compared with groups C and SEV1.Morphological changes were found only in group SEV2.A total of 45 genes were identified to be differentially expressed in group SEV2 as compared with group C.The level of expression of prostaglandin-endoperoxidase synthase 2 gene (Ptgs2),chemokine (C-C motif) ligand 2(CCL2) gene and dual specificity phosphatase1 (Dusp1) gene increased by at least 4 times in group SEV2.Conclusion Sevoflurane can inhibit the cell viability of TM3 mouse leydig cell in concentration dependent manner through abnormal expression of Ptgs2,CCL2 and Dusp1 genes.

Objective To investigate rite effects of loag-term exposure to low-level sevoflurane on reproductive function in mice.Method F0ny male ICR mice,aged 60 d,weighlag 20-25 g,were randomly divided into 4 groups(n=10 each):control group received no sevoflunme(C);group S1-3 were exposed to 0.003%.0.01% and 0.03% sevoflurane 2 h per day for 5 consecutive days per week for 8 weeks respectively. The mice were then sacrificed at the end of the 8 weeks.The testes and epididymis were emoved and sampled for determination of the activities of total lactic dehydregenase(LDH)and lactic dehydrogenase-X(LDH-X),and the motility rate,amount,and aberration rate of sperm.Testicular uhrastructure were observed by transmission electron microscopy.Results The sperm motifity nne were significantly lower.the sperm aberration rate higher and the activity of LDH-X lower in group S3 than in group C(P<0.05),but there was no significant difference in the above parametem between group SI and group S2(P>0.05).The pathology changed of testes occurred only in group S3 among the 3 groups.Conclusion Long-term exposure to 0.03% sevoflurane can result in the abnormality of the reproductive function in male mice but exposure to≤0.01%sevoflurane dose not.

Objective To evaluate the effects of different concentrations of parecoxib sodium on the rat sperm motility,capacitation and acrosome reaction in vitro.Methods The sperm samples from Sprague-Dawley rat epididymis were collected by Klinefelter diffusion method and randomly divided into 4 groups (n =18 each) using a random number table:control group (group C),and parecoxib sodium 100,500,1 000 μmol/L groups (P1-3 groups).Parecoxib sodium with the final concentrations of 100,500 and 1 000 μmol/L was added to the culture medium.The samples were then incubated for 5 h in an airtight container filled with 5 ％ CO2 at 37 ℃.Then sperm motility was examined in vitro at 37 ℃ and analyzed by the computer-assisted sperm analysis,including the sperm motility ((a + b)％),average path velocity (VAP),straight line velocity (VSL),curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH).The capacitation effect was assessed by using the chlortetracycline staining and phase-contract microscopy.The acrosome reaction was evaluated by coomassie brilliant blue staining.Results The VAP,VSL,VCL and capacitation ability of the sperm were gradually decreased in C and P1-3 groups (P ＜ 0.05).Compared with group C,(a + b)％ in P2,3 groups and ALH in P2 group were significantly decreased (P ＜ 0.05).There was no significant difference in the acrosome reaction between groups (P ＞ 0.05).Conclusion Parecoxib sodium has significant inhibitory effects on the rat sperm motility and capacitation in a dose-dependent manner,while has no effect on the acrosome reaction in vitro.

AIM:To evaluate the effect of curcumin on impaired learning-memory ability and the expression of high mobility group box protein 1 ( HMGB1 ) and c-Jun N-terminal kinase ( JNK ) in a rat model of Alzheimer disease (AD).METHODS:Male Sprague-Dawley rats, weighing 250~270 g, were randomly divided into 4 groups (n=9):blank control group (group A), model group (group B), curcumin treatment group (group C, curcumin injected intraper-itoneally at 100 mg· kg-1· d-1 for 6 consecutive days) and solvent control group (group D).The rats of AD model were induced by injection of ibotenic acid into the nucleus basalis of Meynert ( NBM) bilaterally.All rats were trained in Morris maze to assess the ability of learning and memory .The expression of HMGB1 and JNK in the hippocampus was detected by the methods of immunohistochemistry and Western blotting .RESULTS:Compared with group A , the average escape laten-cy (AEL) in groups B and D were obviously longer (P<0.05), while AEL in group C in the 5th and 6th days were signif-icantly shorter (P<0.05).The releases of HMGB1 in the CA1 and CA3 areas in groups B and D from the nucleus were a-bundant.Compared with groups B and D , HMGB1 in hippocampal CA1 and CA3 areas in group C secreted out of the nu-cleus decreased obviously (P<0.05).No significant difference of the release of HMGB1 between group A and group C was observed (P>0.05).No significant difference in the expression of HMGB1 in the hippocampus among the 4 groups was found (P>0.05).However, compared with groups B and D , the expression of JNK in group C was decreased obvi-ously (P<0.05).CONCLUSION: Curcumin significantly improves the learning and memory ability of AD rats .The probable mechanisms may be related to inhibiting the release of HMGB 1 from the nucleus of hippocampal neurons and de-creasing the expression of JNK in the hippocampus .

Objective To evaluate the role of adenosine A1 receptors in hippocampal neurons in the cognitive dysfunction caused by isoflurane anesthesia in aged mice.Methods Sixteen male adenosine A1 receptor gene knockout homozygote mice (gene knockout mice) and 16 male wild-type mice,aged 18-22 months,weighing 27-32 g,were studied.Each type of mice was randomly divided into 2 groups (n=8 each) using a random number table:control group (group C) and isoflurane anesthesia group (group Ⅰ).Mice inhaled 1.4％ isoflurane in 100％ O2 for 2 h in group Ⅰ,and 100％ O2 for 2 h in group C.All the mice underwent Morris water maze test at 24 h after isoflurane or O2 inhalation.After the test,the mice were sacrificed and the hippocampal tissues were harvested to determine the number of β-amyloid1-42 (Aβ1-42) plaques (using immunohistochemistry) and expression of phosphorylated tau (p-tau) protein,and 2B subunit-containing N-methyl-D-aspartate receptors (NR2B) (by Western blot analysis).Results Compared with group C of wild type mice,the escape latency was significantly prolonged,the number of Aβ1-42 plaques was enlarged,the expression of p-tau protein was up-regulated,and the expression of N R2B was down-regulated in group Ⅰ of wild type mice.Compared with group Ⅰ of wild type mice,the escape latency was significantly shortened,the number of Aβ1-42 plaques was decreased,the expression of p-tau protein was down-regulated,and the expression of NR2B was up-regulated in group Ⅰ of gene knockout mice.There was no significant difference in the parameters mentioned above between group Ⅰ and group C of gene knockout mice.Conclusion Adenosine A1 receptors in hippocampal neurons mediate isoflurane anesthesia-induced cognitive dysfunction in aged mice,and the mechanism may be related to promotion of deposition of Aβ,phosphorylation of tau protein and inhibition of activities of NR2B.

OBJECTIVE To investigate the effect of curcumin on proliferation of neural stem cells (NSCs) of rats and the mechanism. METHODS NSCs derived from the forebrain of rat E15 embryos were cultured in vitro and identified by neuroepithelial stem cell protein ( nestin and SOX2) staining. NSCs were treated with curcumin 0.1, 0.5, 2.5, 12.5 and 62.5 μmol.L-1 for 24 h, respectively. The cyto-toxicity was estimated by measuring the release of lactate dehydrogenase(LDH). Cell viability and prolif-eration were analyzed respectively by MTT and BrdU assay. The mRNA expression levels of glucocorti-coid receptor (GR), Stat3, Notch1 and p21 were detected by qRT-PCR. The protein expression levels of total GR, Stat3 and phosphorylated Stat3 were measured by Western blotting. RESULTS The primary neural stem cells were identified as NSCs. Curcumin 12.5 and 62.5 μmol.L-1 had cell cytotoxicity( P<0.05). Cell viability assay indicated that curcumin 0.5 and 2.5 μmol.L-1 enhanced NSCs viability( P <0.05), but in 62.5 μmol.L-1 group the cell cytotoxicity was inhibited(P<0.05). Curcumin 0.1, 0.5 and 2.5 μmol.L-1 increased NSCs proliferation ( P < 0. 05), whereas 12. 5 and 62. 5 μmol.L-1 caused a decrease in NSCs proliferation(P<0.05). The mRNA expression level of GR in 0.5 μmol.L-1 group was significantly reduced( P<0.05). Western blotting analysis revealed that the protein expression of GR, Stat3 and p-Stat3 was inhibited by curcumin in 0.5 μmol.L-1 group(P<0.05). CONCLUSION Curcumin stimulates NSCs proliferation, possibly by inhibiting GR mRNA and related protein expression.

Objective To investigate the effects of inhalation anesthetics on human sperm motility and capacitation in vitro. Methods Sperm samples were obtained from normal adults and prepared with discontinuous percoll gradient centrifugation technique. The samples were incubated for 5 h in an airtight glass container filledwith 5% CO2-95% air at 37 ℃ with or without sevoflurane (SEV 2%, 4% ) or isoflurane (ISO 1.1%, 2.2% ).Then human sperm motility was examined in vitro at 37℃ and analyzed by the computer-assisted sperm analysis (CASA), including sperm motility (a + b)%, curvilinear velocity (VCL), straight line velocity (VSL), averagepath velocity (VAP) and amplitude of lateral head displacement (ALH). The capacitation effect was assessed by using the chlortetracycline (CTC) staining and phase-contract microscopy. Results 2% and 4% SEV significantly reduced (a + b)% , VCL, VSL and VAP in a dose-dependent manner, while only 4% SEV significantly decreased ALH and the capacitation ability of the sperm compared with control group. 2.2% ISO significantly decreased ( a + b)%, VCL, VSL and VAP compared with control and 1.1% ISO group. The capacitation ability of the sperm was significantly decreased by 1.1% and 2.2% ISO as compared with control group. Conclusion Sevoflurane and isoflurane have significant inhibitory effects on human sperm motility and capacitation in a dose-dependent manner. Sevoflurane has stronger inhibitory effect than isoflurane.

Objective To assess whether propofol call induce stable psychic dependence in the rats by self-administration experiment. Methods Twenty-four male SD rats 14 weeks old weighing 240一270 mg were studied. Anesthesia was performed with intraperitoneal injection of 3%sodium pentoharbitsl 40 ms/kg and atropine 03 mg/kg.A catheter wag inserted into the right external jugular vein. Penicillin(100 000 U)0.2 ml wag injected through the external jugular vein for anti-infection and heparin sodium(50U/ml)0.1 ml for anticoagulation. The self-administration experiment of 14 days was started after the 7 days of recovery. All the rats were randomly divided into 4 groups(n=6 each):contontrol group(C),propofol 0.56 mg/kg/l group(P1),propofol 1.00 mg/kg group(P2)and pmpofol 1.70 ms/kg group(P3).The experimental events were controlled by a computer with 50 times of the maximum injection per day.The times ofactive and inactive nose-poke response and times of drug iniection were recorded per day.Results Compared with group C and P1,the times of active nosepoke response and injections were significantly increased in group P2 and P3(P<0.01).The times of active nosepoke response and injections per day were significantly increased in group P3 than in group P2(P(0.01).There was no significant difference in the times of active nose-poke response and injections between group C and P1.There was no significant difference in inactive nose-poke resporme between the 4 groups.And the total daily doses of propofol injected in the last 3 days were significantly increased in a dose-dependent manner.Conclusion Propefol can induce the development of psychological dependence in rata and it is related to the dosage.