Cardiomyopathy, Dilated ; pathology ; Humans ; Infant, Newborn ; Male

Objective To explore the mechanism of the catdioprotection of isoflurane delayed preconditioning on myocardial ischemia reperfusion injury in rabbits.Method Thirty male New Zealand white rabbits,weighing 2.0 to 2.5 kg,were randomly divided into three groups(ten for each group):Control group(group C),I/R group(I/R group) ,2.0% isoflurane group(group S) .Group S was exposed to 2.0% isoflurane-100% oxygen for2 h.Group C and I/R group were exposed 2 h to 100% oxygen served as untreated controls.Twenty-four hours later I/R group and group S underwent 40 rain of coronary occlusion followed by 2 h of reperfusion.Blood samples were taken from arterial line at 20 min before occlusion(T1) ,20 rain after occlusion(T2) ,40 rain after occlusion(T3) ,1 h after reperfusion(T4) and 2 h after reperfusion(TS) for determination of plasma IL-10 levels and TNF-alevels by ELISA.At the end of the reperfusion,infarct size and area at risk were defined by Evans and TTC staining.The heart was harvested and levels of the nuclear factor kappa β(NF-κB)activity were determined by Western Blot,and ultrastructures were observed by electron microscopy.The data was expressed as,and were analyzed by using oneway ANOVA test with SPSS 13.0.P value less than 0.05 indicated statistical significance.Results The NF-κB activity of group S was significantly lower than that of group I/R(P<0.05).Group S significantly(P<0.05)reduced infarct size(19.7%±2.8% in group S) of the left ventricular area at risk as compared with control (37.8 %±1.7 % in I/R group).The injury of I/R group was worse than that of group S from the changes of the cellular structure under light microscope.Group S had a lower levels of TNF-α and also had a higher level of IL-10.Conclusions Isoflurane can inhibit NF-κB activity during myocardial ischemia reperfusion and modulate the cytokine expression,which may be one of molecular mechanisms of Isoflurane delayed preconditioning on cardioprotection.

Objective To investigate the role of opioid receptors in the protective effects of isoflurane-induced delayed preconditioning against myocardial ischemia-reperfusion (I/R) injury in rabbits. Methods Forty male New Zealand white rabbits weighing 2.0-2.5 kg were randomly assigned into 4 groups ( n = 10 each) : group I sham operation (S); group II I/R; group Ⅲ isoflurane + I/R (Iso) and group IV Iso + naloxone + I/R (Nal). Myocardial I/R was induced by 40 min occlusion of left anterior descending branch (LAD) of coronary artery followed by 120 min reperfusion. In group Ⅲ (Iso) 2% isoflurane in 100% O2 was inhaled for 2 h and I/R was produced 24 h later. In group IV (Nal) naloxone 6 mg/kg was given iv 10 min before 2 h of 2% isoflurane inhalation and I/R was produced 24 h later. At the end of 120 min reperfusion, infarct size (IS) and area at risk (AAR) were determined by Evan's blue and TTC staining. Myocardial ultrastructure was examined by electron microscopy. The phosphorylated p38MAPK protein expression in myocardium was determined by Western blot. Results The IS was significantly smaller in group Iso ( Ⅲ ) ( 19.7% ± 2.8%) than in I/R group ( II ) (37.8% ±1.7%) (P<0.05). The phosphorylated p38MAPK protein expression in myocardium was significantly lower in group Iso than in group I/R. Microscopic examination showed less myocardial damage in Iso group than in group I/R. The protective effects of delayed preconditioning by isoflurane was prevented by naloxone pretreatment. ConclusionOpioid receptors may be involved in the protective effects of delayed preconditioning by isoflurane against myocardial I/R injury.

Objective To investigate the effects of isoflurane delayed preconditioning(IDP) on myocardial proteomin rabbits with myocardial ischemia-reperfuaion(I/R).mjury.Methods Eight New Zealand white rabbits of both sexes weighing 2.0-2.5 kg were randomly divided into 2 groups(n=4 each):I/R group and IDP group.Myocardial I/R Wgg induced by occlusion of left anterior descending artery for 40 min followed by 120 min repednsion.In group IDP.the animals inhaled 2%isoflurane for 2 h,undergoing I/R 24 h later.At the end of 120 min reperfusion,the myocardium of left ventricle anterior wall was removed for two-dimensional gel electrophorvsis.The different protein spots were analyzed by means of mass chromatography.Results There were 13 different protein spots between group I/R and IDP.Of the 13 proteins,the expression of 10 spots Was up-regulated and 3 spots down-regulated in quantity.Eleven protein spots of all spots were identified by means of MALDI-TOF-MS.Conclusion IDP can aNenuate myocardial I/R injury in rabbits and it may be related to the alteration in proteome of myocardium.

Objective To investigate the feasibility of macrophages as cell carriers to deliver floate modified oxygen loaded ultrasound contrast agent . Methods The phagocytic activity of macrophages was analyzed by ink phagocytose test , and the expression of folate recepters ( FRs ) on macrophages cell membrane surface was tested by immunofluofluorescence assay . Oxygen/paclitaxel loaded lipid microbubbles( OPLMB) and folate‐targeted OPLMB ( TOPLMB) were synthesized by mechanical shock method and incubated with macrophages in vitro . According to different treatment conditions ,the cells were divided into three groups:group A ( OPLMB) ,group B ( free folic acid + TOPLMB) and group C ( TOPLMB) , the fluorescence intensity of the cells were observed under fluorescence microscope ,and the phagocytic percentage and the phagocytic index of macrophages uptake OPLMB and TOPLMB were observed by bright field microscope . Results The phagocytic percentage of macrophages phagocytose ink was (99 .3 ± 1 .0)% ,FRs was highly expressed on macrophages cultured in vitro . After incubation for 30 minutes ,the fluorescence intensity of group C was significantly higher than those of A and B ,the phagocytic percentage in three groups were (19 .5 ± 0 .2)% ,(21 .0 ± 0 .2)% and (81 .2 ± 10 .0)% respetively . The phagocytic percentage of group C were significantly higher than those in group A and group B ( P <0 .05) . Conclusions Macrophages cultured in vitro possess highly phagocytic activity and these cells highly express FRs ,and can be used as cell carriers to deliver floate modified oxygen loaded multimodality ultrasound contrast agent .

Alteration in the structural appearance of the optic disk and retinal nerve fiber layer usually precedes the development of reproducible glaucomatous visual field defects.Identification of these changes is important in the diagnosis and treatment of glaucoma at an early to moderate stage and in monitoring its clinical course.This paper reviews the optical coherence tomography(OCT),Heidelberg retina tomography(HRT),optic nerve head stereophotograph(ONHP)and scanning laser polarimetry(SLP) in the field of open angle glaucoma.

AIM:To investigate the effect of homeodomain-interacting protein kinase 2 (HIPK2) on the viabi-lity, apoptosis and JAK2/STAT3 signaling pathway in NRK-52E renal tubular epithelial cells induced by hypoxia and reox-ygenation (H/R). METHODS:HIPK2 small interfering RNA (siRNA) was transfected into NRK-52E cells by Lipo-fectamineTM 2000, and normal control group (control group) and negative control group (HIPK2-NC group) were set up. After H/R, the cell viability was measured by CCK-8 assay, the apoptotic rate and Ca2+ fluorescence intensity were ana-lyzed by flow cytometry, and the protein levels of Ki67, cleaved caspase-3, caspase-12, Bcl-2, Bax, p-JAK2 and p-STAT3 were determined by Western blot. RESULTS:Compared with control group, the protein expression of HIPK2 in the NRK-52E cells was significantly decreased after transfection with HIPK2 siRNA (P<0.05). Compared with control group, the cell viability and the protein expression of Ki67 and Bcl-2 in H/R group were also significantly decreased, and the apoptotic rate, the Ca2+ fluorescence intensity and the protein levels of cleaved caspase-3, caspase-12, Bax, p-JAK2 and p-STAT3 were significantly increased (P<0.05). Compared with H/R group, the cell viability and the protein expression of Ki67 and Bcl-2 in HIPK2-siRNA+H/R group were significantly increased, while the apoptotic rate, the Ca2+ fluorescence inten-sity and the protein levels of cleaved caspase-3, caspase-12, Bax, p-JAK2 and p-STAT3 were significantly decreased (P<0.05). CONCLUSION:Inhibition of HIPK2 gene expression promotes H/R-induced growth of NRK-52E renal tubular epi-thelial cells, and reduces the apoptosis. The mechanism is related to down-regulating the JAK2/STAT3 signaling pathway.

Aim To investigate the inhibitory effects of gossypol on migration in gastric carcinoma cell lines and its mechanisms. Methods Gastric carcinoma cells were treated with gossypol at different concentra-tions. The effects of gossypol on cells proliferation were measured using the MTT assay. The migration of gas-tric carcinoma cells was detected by transwell assay. The activation of Akt/β-catenin pathway and the ex-pressions of pathway related proteins ( p-Akt,β-cate-nin, cyclin D1, MMP-2, E-cadherin and vimentin ) were detected by Western blot. Results Gossypol treatment could significantly inhibit the proliferation of gastric carcinoma cells in a dose-dependent manner. Transwell assay showed that the migration ability of
gastric carcinoma cells was significantly decreased. The inhibitory effect of gossypol on cells migration was more significant than the effect of gossypol on cells prolifera-tion. Compared with the control group, treatment with gossypol significantly suppressed the expressions of p-Akt,β-catenin, cyclin D1, MMP-2 and vimentin pro-tein, whereas the expression of E-cadherin was signifi-cantly up-regulated in gastric carcinoma cells in a dose-dependent manner. Conclusion These results demonstrate that gossypol represses cell migration of gastric carcinoma cells through the down-regulation of the activity of Akt/β-catenin pathway.

Objective:To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits, and to explore the possible mechanism. Methods:Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method, with 8 rats in each group. Rabbits in the blank group were fed routinely with normal diet; rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model. Rabbits in the blank and the model groups were not treated. After the model was prepared, rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer; rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively. After 4 weeks of treatment, serum was collected for enzyme-linked immunosorbent assay (ELISA), and the liver tissues were isolated for immunohistochemistry, quantitative polymerase chain reaction (qPCR) and Western-blotting (WB) detection. Results: Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group (P＜0.05); compared with the model group, leptin was significantly increased in the non-transdermal absorption enhancer, the laurocapram and the borneol groups (all P＜0.05); compared with the non-transdermal absorption enhancer group, leptin was significantly increased in the laurocapram group and the borneol group (both P＜0.05); there was no significant difference in leptin between the laurocapram and the borneol groups (P＞0.05). The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin, Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in the model group were significantly lower than those in the blank group (all P＜0.05); compared with the model group, the mRNA expressions of leptin, leptin receptor (LR), JAK2 and STAT3 in the non-transdermal absorption enhancer, the laurocapram and the borneol groups were significantly increased (all P＜0.05); compared with the non-transdermal absorption enhancer group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the laurocapram and the borneol groups were significantly increased (all P＜0.05); compared with the laurocapram group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the borneol group were significantly increased (P＜0.05). The trend of immunohistochemistry and WB detection results was basically consistent with the qPCR assay results. The immunohistochemistry and WB detection results of phosphorylated JAK2 (phospho-JAK2) and phosphorylated STAT3 (phospho-STAT3) were basically consistent with those of JAK2 and STAT3. Conclusion: The molecular expression of Leptin/JAK2/STAT3 pathway in the hyperlipidemia model rabbits was decreased. The molecular expression of Leptin/JAK2/STAT3 pathway was significantly increased after the herbal cake-partitioned moxibustion. The application of laurocapram and borneol, as transdermal absorption enhancers, in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK2/STAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.

Objective:To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. Results:After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (allP<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Conclusion: Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.