f fibrosis-related factors in LX-2cells were significantly increased after co-cultured with QSG7701-HBx cells, which proved that HBx could induce fibrogenesis in vitro.

BACKGROUND:Exercises play an important role in the recovery of neurological function after stroke. Few studies concerned the amount of exercise in rats after stroke. Hippocampus is strongly associated with cognitive function, but no reports addressed the expression of repulsive guidance molecule A in the rat hippocampus after ischemia and reperfusion. OBJECTIVE:To explore the effects of exercise on repulsive guidance molecule A expression in the hippocampus on the ischemic side in rats after cerebral ischemia-reperfusion injury. METHODS:120 Sprague-Dawley rats were equal y and randomly divided into normal group, sham-operation group, and 7-, 14-, 28-day model groups. The model of right cerebral ischemia-reperfusion injury was induced by ligation with nylon monofilament in rats of 7-, 14-, 28-day model groups. Low exercise group received treadmil training of 5 m/min, 5 minutes;7 m/min, 5 minutes;9 m/min, 20 minutes. Moderate exercise group received treadmil training of 8 m/min, 5 minutes;10 m/min, 5 minutes;13 m/min, 20 minutes. High exercise group received treadmil training of 8 m/min, 5 minutes;11 m/min, 5 minutes;20 m/min, 20 minutes. RESULTS AND CONCLUSION:Repulsive guidance molecule A mRNA and protein expression was highest in the ischemic side of the hippocampus in the 7-day model group without excercise. Moreover, repulsive guidance molecule A relative expression gradual y reduced over time. Compared with non-exercise, repulsive guidance molecule A mRNA and protein expression significantly decreased in the 14-and 28-day model groups during moderate exercise (P<0.05), but repulsive guidance molecule A mRNA and protein expression increased during high exercise. Above data confirmed that moderate exercises could decrease repulsive guidance molecule A expression in the affected side of the hippocampus of rats with cerebral ischemia-reperfusion injury.

[Objective]To study the effect of femoral offset and hip joint center on joint function after total hip replacement,radiographic measurements were taken postoperation.[Method]A series of 92 hip joints(87 patients)were followed up.The average follow up priod was 25 monthes.The femoral offset and the position of the prothesis head center were measured in the orthophoric hip joint X-ray photograph and were compared with anatomic Fo and HJC.H arris evaluation system was used to evaluate joint function in four groups.The results were statistically analyzed,with Fisher' exact probability and P value less than 0.05 indicating significant difference.[Result]The coincidence rate of group A(both FO and HJC,27 hips)was 29.35%,group B(only Fo,23 hips)25.00%,group C(only HJC,31 hips)33.70%,group D(neither FO nor HJC,11 hips)11.96%.93.0% patients got the Harris score more than 80 for group A,73.19%(group B),74.19%(group C),27.27%(group D).The difference of Harris evaluation between A and B,A and C,A and D was significant statistically.[Conclusion]Based on the results of the study,the recovery of femoral offset and hip joint center should be considered to contribute to the healing effect after total hip replacement directly.

Objective To investigate the value of soluble triggering receptor expressed on myeloid cells-1(sTREM-1) in the diagnosis of bacterial pleural effusion. Methods The levels of sTREM-1 in pleural effusion were determined in 30 patients with bacterial pleural effusion, 33 patients with malignant pleural effusion,31 patients with tuberculous pleural effusion and 28 patients with transudate by quantitative ELISA assay. The levels of CRP in pleural effusion were assayed using immunonephelometry method. The diagnostic value was assessed by receiver operating characteristic ROC curve analysis. Results The levels of sTREM-1［(1255.2±248.6)ng/L］ in bacterial pleural effusion were significantly higher than those in malignant ［（125.6±22.4）ng/L］, tuberculous［（184.5±36.5）ng/L］ and transudate groups［（92.5±20.8）ng/L］ (P＜0.05). For ROC curve, when the cut off value of sTREM-1 was set at 425 ng/L, the sensitivity , specificity ,accuracy, positive predictive value and negative value was 93.3%,90.3%,91.9%,93.2% and 93.3%, respectively. Conclusions Detection of sTREM-1 in pleural effusion is helpful to differentiate pleural effusion of bacterial origin from those with other etiologies.

Objective To investigate the expression of cytokeratin 34βE12 in esophageal squamous cell carcinoma (ESCC),and its mechanism of action in the process of occurrence and development of an ESCC.Methods Immunohistochemistry was used to analyze the expression of cytokeratin 34βE12 in 252 ESCC patients,66 patients with esophageal carcinoma in situ,and 106 patients with adjacent normal esophageal mucosa before the relationship between its expression and biological behavior was evaluated on the basis of complete clinical information.In addition,Western blotting was used to determine the expression of cytokeratin 34βE12 in 60 patients with esophageal cancer and adjacent normal esophageal tissues.Results (1)The positive rate of caveolin-1 in ESCC,carcinoma in situ,and adjacent normal tissues was 85.7％,54.5％,and 25.7％,respectively.The difference between them was statistically significant (P ＜0.01).(2)The positive rate of cytokeratin 34βE12 in stages Ⅰ,Ⅱ,Ⅲ of ESCC was 76.5％,84.7％,and 96.3％,respectively.The expression intensity of cytokeratin 34βE12 in carcinoma tissue was gradually increased with the advance of clinical stages with a statistically significant difference (P =0.038).The positive rate of cytokeratin 34βE12 with group of lymph node metastasis was significantly higher than those without lymph node metastasis (P ＜ 0.01).(3)Western blotting results further confirmed that the expression of cytokeratin 34βE12 in ESCC was significantly higher than that in adjacent normal esophageal tissue (P ＜0.01).Conclusions The high expression of caveolin-1 might be involved in the occurrence and development of esophageal cancer.The expression of cytokeratin 34βE12 was correlated with the clinical stage of esophageal cancer.cytokeratin 34βE12 was a potential therapeutic target and a valuable prognostic indicator of esophageal cancer progression.

Objective To detect the effect of olfactory bulb(OB)lesion on proliferation,migration and differentiation of the neural stem cells(NSCs)in the subventricular zone(SVZ).Methods Forty-two healthy female SD rats were enrolled and randomly divided into normal control group,isotonic saline group and OB lesion at day 3,at weeks 1,2,3 and 4 groups,six rats per group.OB lesion was induced by N-methyl-D-aspartic acid(NMDA)injection.Bromodeoxyuridine(BrdU)was injected intraperitoneally to label NSCs.Immunohistochemical staining was used to detect the proliferation of SVZ NSCs.In addition,another 18 rats were randomly divided into normal control group,isotonic saline group and lesion group,six rats per group.BrdU was injected intraperitoneally one week after OB lesion and then the animals were sacrificed four weeks after BrdU injection to detect the migration and differentiation of NSCs with immunohistochemistry and immunofluorescence.Results Three days after OB lesion,BrdU-positive cells in SVZ began to increase(26.33 ± 2.58,P ＜0.01),reached the maximum at week 1 (35.33 +3.01,P＜0.01)and still sustained a high level at week 4(19.50+ 2.26,P＞0.05).Five weeks after the OB lesion,the rostral migratory-stream(RMS)and the BrdU-positive cells in OB were significantly increased(86.50 + 5.09,P ＜ 0.01)and(52.83 + 3.87,P ＜ 0.01),respectively.By using fluorescence double staining,most of the BrdU-positive cells were co-localized with the neuronal nuclear antigen(Neun),with a portion of BrdU-positive cells expressing the glial fibrillary acidic protein (GFAP).Conclusions OB lesion can improve the proliferation of NSCs in SVZ and migration of NSCs to OB.The newborn cells can differentiate into not only neurons,but also gliocytes and may be involved in lesion repair.

Objective To study the effects of protein kinase A (PKA) phosphorylation site' s mutation of mitofusin-2 (Mfn2) on intimal proliferation after balloon injury of carotid artery of rats. Method Rat model of carotid artery balloon injury was established and infected with Adv-LacZ, Adv-Mfn2, AdvMfn2-S442A or Adv-Mfn2-S442D from the peri-arterial sheathes of vessels, while phosphate buffered solution (PBS) used instead of above infectious adenovirus as uninfected group and sham operation as control group. The rats were sacrificed 14 days after balloon injury of carotid artery in order to measure the level of Mfn2 protein and the prol9iferation cell nuclear antigen (PCNA) with immunohistochemistry staining. The morphology of vessels was observed with HE staining. All data were statistically analyzed with ANOVA and Dunnett-t test. Results Fourteen days after surgery, the levels of Mfn2 protein significantly increased in arteries infected with Adv-Mfn2, Adv-Mfn2-S4442A and Adv-Mfn2-S442D compared with those in control group, sham operation group and Adv-LacZ infected group. The ratio of intimal area/medial area (I/M) and percentage of PCNA positive cells in both Adv-Mfn2 and Adv-Mfn2-S442A groups markedly decreased compared with control group (P ＜0. 01 ) . Compared with the Adv-Mfn2 group, the I/M and the percentage of PCNA positive cells reduced more significantly in Adv-S442A group (P ＜ 0. 01 ) . However,the I/M and the percentage of PCNA positive cells in Adv-LacZ and Adv-S442D groups were not significantly different from those found in the control group. Conclusions The over-expression of Mfn2 gene may effectively inhibit intimal proliferation after balloon injury of carotid artery of rats. The inhibitory effects of Mfn2-S442A are more obvious than those of Mfn2. However, the Mfn2-S442D is out of the inhibitory effect on neo-intimal proliferation.

BACKGROUND: The mitofusin 2 (Mfn2) may affects vascular smooth muscle cell Ras protein and suppress cellular proliferation through inhibition of extracellular signal-regulated protein kinase signaling pathway, which plays an important role in pathogenesis of vascular disorders such as hypertension, atherosclerosis and post-angioplasty restenosis. Mfn-2 gene amino acid sequence of the first 442 serine serves as protein kinase A (PKA) phosphorylation site, which is closely related to its phosphorylation status and may be involved in its functional regulation.OBJECTIVE: To investigate the effect of Mfn2 gene with PKA phosphorylation site deletion[Mfn2-PKA (△)] on inhibiting the proliferation of vascular smooth muscle cells and related signaling pathway.METHODS: Vascular smooth muscle cells of rats infected by recombinational adenovirus carrying green fluorescent protein,Mfn2 gane and Mfn2-PKA (△), were subcultured for 3-10 passages and randomly divided into 4 groups: ① Control group without intervention. ② Control group infected with adenovirus carrying green fluorescent protein. ③ Experiment group infected with adenovirus carrying Mfn-2 gene.④ Experiment group infected with adenovirus carrying Mfn2-PKA (△). Laser confocal microscopy was used to observe the locations of Mfn2 gene with and without PKA in cells. The expressions of extracallular signal-regulated protein kinase, Mfn2 gone and Mfn2-PKA (△) were determined by Western blot analysis. The growth curve of the vascular smooth muscle cells was explored by MTT.RESULTS AND CONCLUSION: The Mfn-2 and Mfn2-PKA (△) both expressed protein-specific bands in vascular smooth muscle cells. Two kinds of gone expression products were mainly located at the out membrane of mitochondria. Compared with the control group and adenovirus carrying green fluorescent protein group, the absorbance values at 3, 4, 5, 6 days were significantly reduced in adenovirus carrying Mfn2 group (P ＜ 0.01), and no obvious changes were observed in adenovirus carrying Mfn2-PKA (△) group. Compared with the control group and adenovirus carrying green fluorescent protein group, the extracellular signal-regulated protein kinase expression was significantly reduced in adenovirus carrying Mfn2 group (P ＜ 0.01), and no obvious changes were observed in adenovirus carrying Mfn2-PKA (△) group. Mfn2-PKA (△) located at the out membrane of mitochondria, has no effect on suppressing the proliferation of vascular smooth muscle cells, and no inhibition effect on extracellular signal-regulated protein kinase signaling pathway.

BACKGROUND: Mitofusin2 (Mfn2) profoundly inhibits proliferation and induces apoptosis of vascular smooth muscle cells (VSMCs) through phosphatidylinositol 3 kinase/protein kinase B. Notably, tMfn2, with the transmembrane domain deleted, has a 41%-reduced molecular weight, which possibly exhibits a stronger effect on inducing apoptosis.OBJECTIVE: To investigate the effect of rat tMfn2 gene, with the transmembrane domain deleted, on promoting the apoptosis of VSMCs, and to determine related signal pathway.DESIGN, SETTING AND TIME: A controlled observational study at a gene level was performed in the central laboratory, Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology between January and October in 2008.MATERIALS: Rat VSMCs and the recombinant adenovirus containing LacZ, Mfn2 or tMfn2 was offered by Professor Chen as a gift.METHODS: The rat VSMCs cultured at 3-10 passages were divided into 4 groups. ①Blank control group: No interventions. ② VSMCs were infected by adenovirus-mediated LacZ, Mfn2, and tMfn2, respectively.MAIN OUTCOME MEASURES: ①Expressions of Mfn2 and tMfn2 following the VSMCs were infected with recombinant adenovirus for 24 hours. ②The apoptosis of VSMCs was determined with flow cytometry and enzyme linked immunosorbent assay at 24, 48 and 72 hours following infection. ③Western blot was used to analyze the expression of phosphorylated AKT following the VSMCs were infected with recombinant adenovirus for 24 hours.RESULTS: ①Both Mfn2 and tMfn2 were expressed in the VSMCs. ②The tMfn2 was superior to Mfn2 in promoting the apoptosis of VSMCs in a time-dependent manner (P<0.01). ③The protein expression of phosphorylated AKT remarkably decreased two groups, especially significant in tMfn2-infected group (P<0,01).CONCLUSION: The tMfn2 can induce the apoptosis of VSMCs more effectively via the inhibition of phosphorylated AKT signaling pathway.

BACKGROUND:So far the positive or negative effects of mesenchymal stem cel s on tumor growth and metastasis are under discussion. OBJECTIVE:To explore the mechanism of bone marrow mesenchymal cel s in promoting lung cancer metastasis. METHODS:Primary rat bone marrow mesenchymal stem cel s were obtained by direct adherence method of the whole bone marrow, and differential adherence combined with digestion control method was performed to purify cel s. Lung cancer cel lines were cultured, and the effects of bone marrow mesenchymal stem cel s on the migration, invasion and metastasis of lung cancer cel s were observed by scratch test, cel invasion and migration assays. Orthotopic lung cancer models were established in rats and bone marrow mesenchymal stem cel s were seeded onto the left lung of rats. Then, pathological changes of lung tissues were observed at 14 days after transplannation. RESULTS AND CONCLUSION:After the scratch test, the migration rate of lung cancer cel s became higher, and the scratches healed with time. And after cel transplantation, the number of migrated lung cancer cel s increased, as wel as the ability of lung cancer cel s penetrating the Matrigel was strengthened. Besides, fibrous connective tissues could be found around the lung cancer tissues, and necrosis with distinct boundary and large tumor nuclei;the metastatic tissues showed obvious infiltration and necrosis with large tumor nuclei. These results suggest that bone marrow mesenchymal stem cel s can promote the invasion, migration and metastasis of lung cancer cel lines.