After blast injury, the morphologie and ultrastructural differentiation of the rabbit corneal endothelium in primary culture was evaluated and compared with the Na/K ATPase activity that occoured during wounding and healing. Morphologie and ultrastructural changes were investigated by light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The endothelial injury and recovery after blast can be described as a three—stage process. Stage one was characterized by a desquamation, degeneration and necrosis of endothelial cells and a low activity of Na/K ATPase; In stage two, some surviving endothelial cells appeared to migrate and divide and the activity of Na/K ATPase began to increase. In stage three, surviving endothelial cells displayed a flattened configuration consisting of polygons and a high ATPase activity.

Objective To investigate the effect of different doses of lidocaine on expression of the high mobility group box 1 protein (HMGB1) in small intestine in septic rats.Methods Fifty adult male Wistar rats weighing 200-250 g were randomly divided into 5 groups ( n =10 each):sham operation group (group S),sepsis group( group Sep ) and different doses of lidocaine group (group L1～3 ).Group S were not applied cecal ligation and puncture (CLP).Sepsis intestinal damage model was performed in group Sep by CLP.Group L1～3 were given intraperitoneally lidocaine in a dose ofS,10 and 20 mg/kg at immediately,1 and 2 h after CLP,respectively.Isometric normal saline was given intraperitoneally in group S and group Sep.The small intestine tissues were taken at 24 and 48 h after CLP.The small intestine morphology was observed with optical microscope.The expression of the HMGB1 mRNA were examined by PCR and the activity of diamine oxidase (DAO) were determined by ELISA.Results Compared with group S,the expression of HMGB1 mRNA was increased and the activity of DAO decreased in group Sep and groups L1～3 at 24 and 48 h after CLP ( P ＜ 0.05 ).Compared with group Sep,the expression of HMGB1 mRNA was decreased and the activity of DAO increased in a dose-dependent manner in groups L1～3 ( P ＜0.05),The injury of pathology of small intestine was slighter in groups L1～3 than in group Sep.Conclusion Lidocaine can reduce samll intestine injury through inhibiting HMGB1 expression in septic rats by a dose-dependent manner.

To compare the clinical effect of needling the Ashi point (4cm lateral to the lower border of the spinous process of cervical vertebra) plus Yaotongdian on the hand and point injection on the Ashi point with Triamoninde. The curative effect of acupuncture was better than that of point injection.

To accurately analyze metabolites in industry-important photosynthetic microbes, LC-MS based metabolomics protocol needs to be optimized specifically for individual species. In this study, an LC-MS based metabolomics method was optimized for cyanobacterium Synechocystis sp. PCC 6803. With the optimized extraction, liquid chromatographic and mass spectral parameters, the method was capable of detecting 24 important metabolites related to central carbohydrate and energy metabolism in Synechocystis sp. PCC 6803. The study laid an important foundation for the metabolomics analysis of cyanobacteria.
Chromatography, Liquid ; Mass Spectrometry ; Metabolome ; Metabolomics ; Photosynthesis ; Synechocystis ; metabolism

Objective: Our purpose was to study the relationship between the Fas antigen expression and neuron apoptosis after cerebral ischemia. Methods: We detected the Fas antigen expression and neuron apoptosis dynamically in the animal models with cerebral ischemia by immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. Results: Fas antigen was positive after 30 minutes of ischemia and reached peak at 60 minutes. At the 24th hour, it began to decrease, and negative on the 3th day. While the positive cell for TUNEL method appeared after 60 minutes of ischemia, reached peak on 3 day, and decreased on 7 day. Conclusion: Neuron apoptosis after cerebral ischemia is closely related to the over-expression of Fas antigen.

Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.

Objective To study the expression of VEGF-C and it's relationship with pathologic characteristics in rectal cancer.Method In this study 82 patients of mid to low rectal cancer underwent radieal resection from December 2007 to August 2008.Tumor tissues and lymph nodes were studied by immunohistochemical staining for VEGF-C expression in the tumor tissue and CK20 expression in D3 lymph nodes.Results were compared with the pathological results of mesenterium lymph nodes to explain the relationships between VEGF-C expressions and clinical pathologic characteristics.Data were analyzed with Chi square test.Result As for the expression of VEGF-C.significant differences were found between tumor stages(X~2=8.529,P<0.05)and between positive and negative lymph node metastasis(X~2=4.712,P<0.05).but there was no difference between that of mesenterium lymph node metastasis and D3 lymph node micrometastasis(X~2=0.017,P>0.05). Conclmion In rectal cancer,VEGF-C expression is correlated with tumor stage,type,metastasis and micrometastasis of lymph node.

Objective To investigate the effects of different CO2 pneumoperitoneum pressures on gastric cancer cells' growth and proliferation in nude mouse model of implanted tumor. Methods Human gastric cancer cell lines MNK-45 were exposed under 0、10、12 and 15 mm Hg CO2 pneumoperitoneum for 4 hrs respectively. 2 × 106 processed cells were inplanted into nude mice subcutaneously. Three weeks later, mice were sacrificed and the weight and bulk of the tumor measured. Then we observed the transplantation tumor by HE stain and Ki-67 stain. Results There was no significant difference in tumor's growing time, bulk and weight between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (7. 8 d, 7. 2 d, 7. 8 d; 1. 2 cm3, 1. 3 cm3, 1. 3 cm3; 1.5 g, 1. 9 g, 1. 6 g)and the control group (7. 3 d, 1. 2 cm3, 1.4 g) (P > 0. 05 ). The growing time of tumor in 15 mm Hg CO2 pneumoperitoneum (12. 5 d) was obviously longer than the control group ( P < 0.05 ) , the bulk and weight of tumor in 15 mm Hg CO2 pneumoperitoneum (0. 5 cm3, 0. 5 g) group significantly decreased compared with the control group (P <0.05). The positive rate of Ki-67 in 15 mm Hg CO2 pneumoperitoneum (27. 5% ) group was obviously lower than the control group (59.6%) (P<0.01). However, there were no significant differences between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (61.2%, 60.5%, 63.4%) and the control group (P > 0.05). Conclusion Clinically adopted CO2 pneumoperitoneum pressures have no significant effect on gastric cancer cells growth and proliferation.

Objective To investigate the effects of exosomes from cultured human retinal pigment epithelium (ARPE-19) cells affected by oxidative stress on the proliferation and expression of vascular endothelial growth factor-A (VEGF-A) and Akt of ARPE-19 cells. Methods Culture ARPE-19 cells. The concentration of 2.5μmol/L rotenone was selected to simulate oxidative stress and isolated ARPE-19-exosome. Exosomes were isolated by ExoQuick exosome precipitation solution. Transmission electron microscopy was used to identify the morphology of exosomes. Western blot was used to detect exosomes’ surface-specific maker protein CD63. ARPE-19 cells affected by oxidative stress were cultured with exosome as experimental group, normal ARPE-19 cells were cultured with exosome as control group. The cell proliferation was examined by methyl thiazolyl tetrazolium assay. Western blot and immunofluorescence assay were used to detect the expression levels of VEGF-A and Akt protein. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the levels of VEGF-A mRNA and Akt mRNA. Results The diameter of normal ARPE-19-exosomes ranged from 50 to 150 nm. The isolated exosomes expressed CD63. AREP-19 cells were cultured with ARPE-19 (affected by rotenone)-exosome, the cell viability in experimental group was significantly reduced than in the control group. Green fluorescence was observed in the cytoplasm under fluorescence microscope. Compared with the control group, VEGF-A was up-regulated expressed and Akt was down-regulated expressed. Western blot results showed that, VEGF-A protein expression in the experimental group were higher than the control group. Akt protein expression in the experimental group were less than the control group. The difference was statically significant (t=3.822, 6.527;P<0.05). RT-PCR results showed that VEGF-A mRNA expression levels was higher in the experimental group than the control group. Akt mRNA expression levels was lower in the experimental group than the control group. The difference was statically significant (t=8.805,?7.823;P<0.05). Conclusions Exosomes from ARPE-19 cells affected by oxidative stress inhibit the proliferation of normal ARPE-19 cells, increase the expression of VEGF-A and reduce the expression of Akt.

Objective To detect the expression of tissue inhibitor of metalloproteinase-4 (TIMP-4) in cutaneous malignant melanoma (CMM) tissue and to assess its relationship with melanoma proliferation,invasion and metastasis.Methods Western blot was conducted to measure the protein expression of TIMP-4 in five fresh lesional and paratumoral tissue specimens of CMM and three fresh tissue specimens of nevi.Immunohistochemistry was carried out to quantify the expression of TIMP-4,Ki-67,matrix metalloproteinase-2 (MMP-2),vascular endothelial growth factor (VEGF) and CD63 in paraffin-embedded tissue samples from 43 cases of CMM and 51 cases of nevi.The degree of malignancy of melanoma was evaluated in these lesions.Results Western blot analysis showed that the expression of TIMP-4 was significantly higher in 4 of 5 CMM tissue specimens than in corresponding paratumoral tissue specimens and nevus tissue specimens.Immunohistochemistry revealed that the expression rate of TIMP-4 was 86.04％ (37/43) in melanoma tissue,compared to 19.6％ (10/51) in nevus tissue (x2 =31.55,P ＜ 0.05).The expression of TIMP-4 increased sequentially from in situ melanoma to invasive and metastatic melanoma (rs =0.309,P ＜ 0.05).As far as CMM was concerned,the TIMP-4 expression was uncorrelated with any of the known prognostic variables including clinical stage,Clark level,Breslow depth,presence of ulcer,and Ki-67 expression (all P ＞ 0.05),but positively correlated with the expressions of VEGF (rs =0.345,P ＜ 0.05) and CD63 (rs =0.555,P ＜ 0.01).The median expression level of TIMP-4 was significantly higher in MMP-2-positive than in MMP-2-negative melanoma tissue samples (3 vs.0,P ＜ 0.01).Conclusions TIMP-4 protein is highly expressed in CMM tissue,which may be closely associated with the initiation and progression of CMM,especially with the metastasis of and angiogenesis in CMM.