AIM:To study the effects of methylene blue (MB) on myocardial ischemia/reperfusion (I/R)-induced mitochondrial injury in isolated rat hearts.METHODS:Spragure-Dawley (SD) rats were divided into 3 groups randomly (n=6): control group, I/R model group and MB treatment group (IR+MB group).The isolated rat hearts were prepared and set up to Langendorff perfusion.The rats in I/R+MB group received MB (2 mg/kg) by intraperitoneal injection 2 h before operation.The hearts in control group were perfused with K-H solution for 110 min consecutively.The hearts in I/R group and I/R+MB group were in equilibrium for 20 min, following by 45 min of global ischemia, and then reperfused for 60 min.The heart rate (HR), left ventricular developed pressure (LVDP), left ventricular pressure maximum change rate (±dp/dtmax) and left ventricular end-diastolic pressure (LVEDP) were recorded.The perfusate was collected to determine the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH).The contents of reactive oxygen species (ROS), malondialdehyde (MDA) and adenosine triphosphate (ATP), and the activity of superoxide dismutase (SOD) in the myocardial tissues were all determined.Histopathological examination of left ventricle was performed.The mitochondria from the heart tissues was isolated and the mitochondrial swelling and mitochondrial membrane potential (MMP) were analyzed.RESULTS:Compared with control group, the hearts in I/R group showed poorer function, higher CK-MB and LDH levels in the perfusate, increased ROS and MDA contents, higher SOD activity and less ATP content in the heart tissues (P<0.05).Furthermore, the mitochondrial swelling level increased and MMP reduced in I/R group (P<0.05).Compared to I/R group, MB improved heart function and reduced the release of CK-MB and LDH (P<0.05).MB also decreased ROS and MDA contents, and increased the activity of SOD and the content of ATP (P<0.05).In addition, MB alleviated mitochondrial swelling and restored the reduced MMP (P<0.05).CONCLUSION: MB protects the isolated rat hearts from I/R-induced injury by attenuating the damage of mitochondria.

AIM: To verify the role of enhancing or suppressing the expression of glutathione peroxidase 1 (GPx1) in the growth, migration and invasion of glioblastoma multiforme cell lines U87MG and U118MG.METHODS:U87MG and U118MG cell lines were transfected with the vector containing specific siRNA or pcDNA3.1 recombinant plas-mid both targeting GPx1.The mRNA and protein expression levels of GPx1 were detected by real-time PCR and Western blotting.MTS assay was applied for determining the cell activity.The abilities of migration and invasion were examined by Transwell assay.RESULTS:Compared with blank control group and negative group, the inhibitory rate of the cell activity in U87MG cells in siRNA group was significantly reduced by 25.9%, 35.7%and 34.8%at 24 h, 48 h and 72 h, respec-tively (P<0.05).In contrast, the cell activity of U118MG cells in pcDNA3.1-GPx1 group was significantly increased by 22.7%, 45.8%and 39.8%at 24 h, 48 h and 72 h, respectively ( P<0.05) .In siRNA group, the inhibitory rate of mi-gration in U87MG cells was 41.6%±8.2%and the invasion was 41.6%±8.2%compared with blank control group and negative group (P<0.05).The cell migration and invasion rates of the U118MG cells in pcDNA-GPx1 group were in-creased by 55.8%±9.8% and 60.8% ±9.2%, respectively, compared with blank control group and negative group (P<0.05).CONCLUSION:The down-regulation of GPx1 by specific siRNA reduces the capability of cell growth, mi-gration and invasion of U87MG cells, while up-regulation of GPx1 by pcDNA3.1-GPx1 increases the capability of cell growth, migration and invasion of U118MG cells.

AIM: To investigate the effects of caveolin-1 (Cav-1) scaffolding domain peptide, cavtratin, on lipopolysaccharide (LPS)-induced mouse acute lung injury and heme oxygenase-1 (HO-1) activity.METHODS: Adult male BALB/c mice were randomly divided into 6 groups (n=8 to 10): control, Antennapedia internalization sequence (AP), LPS, LPS+hemin, LPS+ hemin+cavtratin and LPS+hemin+cavtratin+zinc protoporphyrin IX (ZnPP) groups.After LPS administration for 24 h, the lung pathological changes, the wet/dry weight (W/D) ratio of lung tissues, total cell number in bronchoalveolar lavage fluid and serum lactate dehydrogenase activity were measured.The co-localization of HO-1 and Cav-1 was displayed by immunofluorescence, and the HO-1 activity were detected.The mRNA expression of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, MCP-1 and iNOS was detected by real-time PCR.RESULTS: The mice in LPS+hemin+cavtratin group had the decreased interaction between HO-1 and Cav-1, and the increased HO-1 activity compare with LPS group (P<0.05).Compared with LPS group, the pulmonary damage was attenuated in LPS+hemin+cavtratin group, and the injury indexes, including W/D ratio, total cell number in bronchoalveolar lavage fluid and lactate dehydrogenase activity in the serum, and the mRNA expression of inflammatory cytokines all decreased (P<0.05).HO-1 activity inhibitor ZnPP abolished the above protective effect of cavtratin on the lung tissues with LPS-induced acute lung injury.CONCLUSION: Cavtratin has beneficial effects on the lung with LPS-induced acute injury by restoring the HO-1 activity.

Objective To explore microsurgical treatment of tuberculum sellae meningiomas.Methods A retrospective analysis was made on 35 cases of tuberculum sellae meningiomas operated from January 2005 to July 2013 in neurosurgery department of Sun Yat-sen Memorial Hospital,surgical approach,removal rate,surgical effect and complications were analysed.Results All patients were accepted microsurgical treatment,twenty cases were operated via subfrontal approach,four cases via anterior interhemispheric approach,ten cases via pterional approach,one case via combined subfrontal and pterional approach.According to Simpson grade,grade Ⅱ,rection was achieved in 26 cases,grade Ⅲ in 4 cases and grade Ⅳ in 5 cases.The total rection rate was 85.7％.There were 28 cases with merger ision loss and visual field defects preoperate,twenty cases were improved after operation,five cases with no change,three cases aggravated.The visual improved rate was achieved 71.4％,there was no surgical mortality case.Conclusion The surround tissue of tuberculum sellae meningiomas is very import ant,microsurgical rection is the main treatment.The choice of surgical approach should according to tumor size,growth pattern,degree of impaired vision and surgeon experience.Family with microanatomy and skillfull microsurgical techique can make sure operation succes.

BACKGROUNDTo explore a rapid and accurate method for examining cancer cell from the sputum in patients with lung cancer.

METHODSOne hundred and twenty patients with lung cancer diagnosed by operation and pathologically confirmed were enrolled in this study. Sputum sediment section and sputum smear examinations were performed and compared for diagnosis of lung cancer.

RESULTS(1) The positive rate of lung cancer cell was 71.67% (86/120) by sputum sediment section examination, however, only 31.67% (38/120) by sputum smear examination ( P < 0.001). The diagnostic rate of combination of two methods for lung cancer was 90.83% (109/120), which was significantly higher than that of single sputum sediment section examination ( P < 0.001). (2) With routine HE staining, 55 patients (55/86,63.95%) could be histologically identified by sputum sediment section examination, but only 6 patients (6/38,15.79%) by sputum smear examination ( P < 0.001). (3) In 31 patients unidentified with routine HE staining, 29 were further histologically confirmed by sputum sediment section examination with immunohistochemistry.

CONCLUSIONSCompared to sputum smear examination, sputum sediment section examination can make use of more sputum materials, show a higher sensitivity for cancer cell, and accurately identify the histological classification of tumor. It is supposed to be a good examination for lung cancer and deserved to be extended in clinical application.

Objective To investigate the effect of caveolin-1 scaffolding domain (CSD) peptides on heme oxygenase-1 (HO-1) activity increasing and M1/M2 phenotype polarization in rat alveolar macrophages (AMs) induced by lipopolysaccharide (LPS).Methods Bioinformatics was used to analyze the binding of full-length wild-type CSD polypeptide and 101 amino acid deleted truncated mutant CSD polypeptide (Δ101CSD) to HO-1. Primary AMs were isolated from rats, when cell fusion reached 80％, they were synchronized with serum-free medium and divided into five groups: no treatment was given to the blank control group; LPS group was treated with 100μg/L LPS for 16 hours;LPS+ hemin group was treated with 100μg/L LPS and 20μmol/L hemin for 16 hours; wild-type CSD polypeptide+ LPS+hemin group was pretreated with 10μmol/L wild-type CSD polypeptide 6 hours before LPS treatment; Δ101CSD+ LPS+hemin group was pretreated with 10μmol/L Δ101CSD polypeptide 6 hours before LPS treatment. After treatment for 16 hours, the co-localization between caveolin-1 (Cav-1) and HO-1 was displayed by confocal microscope; the mRNA expressions of inflammatory cytokines interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) and M1/M2 polarization cytokines tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), leukocyte differentiation antigen 206 (CD206) and IL-10 were determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR); the HO-1 activity and nitric oxide (NO) production were determined by spectrophotometry.Results Bioinformatics analysis showed: both wild-type CSD and Δ101CSD peptides could bind to HO-1, and there was no significant difference in the binding ability between the two peptides, but the deletion of 101 Arg resulted in the disappearance of part of the binding region between Δ101CSD and HO-1. The results of laser confocal microscopy showed: the expressions of Cav-1 and HO-1 were lowed in the blank control group, and Cav-1 was bound to HO-1 in LPS group and LPS+ hemin group. Both wild-type CSD and Δ101CSD peptides pretreatment could significantly reduce the binding of HO-1 to Cav-1 induced by LPS. HO-1 activity analysis showed: after LPS stimulation, the activity of HO-1 was significantly higher than that of the blank control group; the activity of HO-1 induced by LPS was increased by hemin; after pretreatment with two kinds of CSD peptides, the activity of HO-1 was further increased, and the effect of wild-type CSD peptide was more significant, which showed a statistically significant difference as compared with that of LPS+ hemin group (pmol·mg-1·h-1: 3683±266 vs. 2408±132,P < 0.05). RT-qPCR results showed: LPS could induce elevation of cytokines and M1 markers and decrease of M2 markers, while hemin could inhibit LPS-induced inflammatory response and M1/M2 phenotypic polarization. Compared with LPS+ hemin group, after pretreatment with wild-type CSD peptide, the levels of inflammatory factors in AMs were decreased, and the mRNA expression levels of TNF-α and iNOS, M1 markers, were decreased [TNF-α mRNA (2-??Ct): 6.82±0.05 vs. 8.70±0.24, iNOS mRNA (2-??Ct): 331.50±32.05 vs. 506.70±0.10, bothP < 0.05], and IL-10 mRNA expression level was increased (2-??Ct: 269.09±6.54 vs. 119.05±3.30,P < 0.05). The deletion of 101 site partially weakened the inhibitory effect of CSD peptides on inflammatory factors and only reduced the expression of iNOS mRNA (2-??Ct: 429.11±8.92 vs. 506.70±0.10,P < 0.05), indicating that its ability to transform AMs from M1 phenotype to M2 phenotype was poor. The two peptides had no effect on the expression of CD206.Conclusion Wild-type CSD had beneficial effects of anti-inflammation by reducing Cav-1 binding to HO-1 induced by LPS, restoring the HO-1 activity and driving M2 phenotype in alveolar macrophages.

Objective: To explore the application value of CT imaging in differentiating gastric stromal tumors (GST) from gastric leiomyomas (GLMs). Methods: CT images of patients with GST (n=65) or GLMs (n=13, maximum diameter of tumor ≤5 cm) proved by surgery and pathology were retrospectively analyzed. The tumor size, location, contour, growth pattern, degree and pattern of enhancement, calcification, necrosis, surface ulceration, lymph nodes, and patient clinical data were evaluated by two independent reviewers. Receiver operating characteristic (ROC) curves were employed to assess the measurement and calculation parameters in the differentiation of GST and GLMs. Results: Between the GST and GLMs groups, there was no statistically significant difference in the contour, growth pattern, calcification, surface ulceration, and patient's sex (P>0.05). CT values of in plain scans, degree of enhancement in arterial phase (DE1), size, location and pattern of enhancement were found to be different between GST and GLMs (P<0.05). When the cutoff value of the maximum tumor diameter was 3.2 cm, the area under ROC curve, sensitivity and specificity were 0.707, 92.3%(12/13) and 60.6%(40/66), respectively. When the cutoff value of age was 59 years, the area under ROC curve, sensitivity and specificity were 0.773, 92.3% (12/13) and 46.2% (30/65), respectively. Taking the cutoff value of 10.9 HU as the degree of enhancement in arterial phase (DE1), the area under ROC curve, sensitivity and specificity were 0.774, 84.6% (11/13) and 77.3% (51/66), respectively. Using a cutoff value of 30.3 HU, the sensitivity, specificity, and the area under ROC curve were 84.6% (11/13), 65.2% (43/66), and 0.731, respectively. Conclusions CT examination in addition to clinical data can be very helpful for the differential diagnosis of GLMs from GSTs in maximum diameter ≤5 cm.