Objective:To study the inhibitory effects of adenovirus mediated vascular endothelial cell growth inhibitor 151(VEGI_(151)) on venous endothelial cell proliferation. Methods: VEGI_(151) gene was cloned into adenovirus vector pCA13 and the product was packaged and amplified with 293A cells.The expressed protein was identified by Western blot. X-gal staining was used to determine the transfection efficiency of the recombinant adenovirus system. The inhibitory effect of Ad-VEGI_(151) on ECV304 cell proliferarion was observed and assessed by violet crystal assay. The protein expression of VEGI_(151) gene in ECV304 cells was detected by immunohistochemistry. Results: Liposome-mediated pCA13-VEGI_(151) and pJM17 cotransfection of 293A cells (through homologous recombination of intracellular plasmid) is a practical and feasible method to obtain high-titre recombinant adenovirus. The constructed Ad-VEGI_(151), with high transfection efficiency, significantly inhibited ECV304 cells proliferation and expressed functional protein in target cells. Conclusion: Ad-VEGI_(151) can express bioactive protein in target cells and can inhibit the proliferation of venous endothelial cells, and this may pave a new way for gene therapy of tumors and neovascular diseases.

As a multifunctional cytokine,transforming growth factor-?(TGF-?) has many biological effects. It controls the extracellular matrix synthesis in trabecular meshwork cells and enhances the contractility of trabecular cells,which may increase the resistance of aqueous outflow. It can also promote the formation of the bleb scarring after filtration surgery. TGF-? is likely to play an important role in the pathogenesis and mechanism of primary open-angle glaucoma (POAG). The inhibitions of TGF-? may be used as an auxiliary agent for inhibiting the formation of the bleb scarring after filtration surgery.

Esophageal Neoplasms ; pathology ; Esophagus ; pathology ; Humans ; Lipoma ; pathology

AIM: To investigate the crosstalk between angiotensin Ⅱ (AngⅡ)-mediated and platelet-derived growth factor (PDGF)-mediated signal transduction in vascular smooth muscle proliferation.METHODS: A model of renal hypertension was made by two kidney/one-clip operation. Level of PDGF receptor β subunit of aorta was measured by Western Blot analysis. The effect of Ang Ⅱ on PDGF receptor β subunit expression was investigated in culture rat aortic vascular smooth muscle cells (VSMC).RESULTS: Systolic blood pressure obviously increased at 8th week after operation, whereas the level of PDGF receptor β subunit of aorta significantly increased by 126.6% (P＜0.05) in 2K1C rats compared with control group. The expression of PDGF receptor β subunit in cultured VSMC stimulated by AngⅡ was higher than that of control by 192.74%(P＜0.01). The effect of AngⅡ was inhibited remarkably by pretreated with losartan, a kind of specific AngⅡ receptor 1 (AT1) subtype antagonist and U73122, a kind of phospholipase C inhibitor. The effect was partly blocked by PD98059, which inhibit the activity of mitogen-activated, ERK-activating kinase (MEK).CONCLUSION: AngⅡ-induced PDGF receptor β subunit expression is regulated by the AT1 and its downstream signal molecule-PLC and ERK, might participate in the intracellular signal transduction pathway.

Objective To explore the proportional changes of regulatory B lymphocyte cells (Bregs) and suppressor T lymphocyte cell (Ts) in the peripheral blood of patients with Hashimotos' thyroiditis (HT).Method Sixty-one cases of HT and 38 healthy subjects with matched age and gender were recruited.Flow cytometry technology was used to detect the proportional changes of CD19+ CD24+ CD27 + B cells,CD19+ CD24hi CD38hi B cells,CD19+ IL-10+B cells,and CD8+CD28-T cells.Result Compared with control group,CD19+CD24+CD27+B cells in HT group increased significantly(P＜0.05).With the increase of thyroid peroxidase antibody,CD19+ CD24hi CD38hi B cells were decreased significantly.With the increase of antithyrogloblin antibody (TGAb),CD8 + CD28-T cells were decreased markedly (P＜0.05).TT3 was positively related with CD19 + CD24hi CD38 hiB cells while TT4 had a negative correlation with CD8+ CD28-T cells and positive correlation with CD19+ CD24+ CD27+ B cells.Logistic regression analysis showed that TGAb and FT4 were independent risk factors for decreased CD8+ CD28-T cells in lymphocyte percentage(P＜0.05).TSH was an independent risk factor for the decreased proportion of CD19+ CD24hi CD38hiB cell.FT4 was the independent risk factors for increased CD19+CD24+CD27+B cells(P＜0.05).Conclusion The percentage of Ts cells in patients with HT was lowered,while the different phenotypic Bregs cells showed various changes.Thyroid autoantibodies and thyroid function were closely associated with the immunological changes in HT patients.

Objective To investigate the alteration of the cytoskeleton of airway smooth muscle cells in young asthmatic rats with airway remodeling and the effect of RhoA/ROCK signal pathway.Methods Airway smooth muscle cells (ASMCs) were primary cultured and purified from Sprague-Dawley(SD) rats that were induced by ovalbumin (OVA) inhalation for 8w,then incubated by Pho kinase inhibitor Y27632.Real time quantitative polymerase chain reaction (RT-PCR),Western blot,and immunohistochemistry were used to measure the alteration of F-actin,and α-tubulin in the cytoskeleton of airway smooth muscle.Results (1) The asthma group showed a high average gray value of F-actin in ASMC than control groups,especially 8 weeks;and were significantly down in the group after adding Y27632(P ＜0.01).(2) The intension and intensity of fluorescence signal of α-tubulin in asthma groups in 8 weeks were higher than control greup(P ＜0.01),and were significantly decreased in Y27632 group.(3) A higher expression of α-tubulin protein was shown in the asthma group in 8 weeks relative to control group(P ＜0.01),and was significantly down-regulated in Y27632 group(P ＜0.05).Conclusions Alteration of the cytoskeleton of airway smooth muscle exists in young asthmatic rats and the RhoA/ROCK signal pathway possibly plays a significant role.

AIM: To investigate the crosstalk between angiotensin Ⅱ (AngⅡ)-mediated and platelet-derived growth factor (PDGF)-mediated signal transduction in vascular smooth muscle proliferation.METHODS: A model of renal hypertension was made by two kidney/one-clip operation. Level of PDGF receptor ? subunit of aorta was measured by Western Blot analysis. The effect of Ang Ⅱ on PDGF receptor ? subunit expression was investigated in culture rat aortic vascular smooth muscle cells (VSMC).RESULTS: Systolic blood pressure obviously increased at 8th week after operation, whereas the level of PDGF receptor ? subunit of aorta significantly increased by 126.6% ( P

This study examined in vitro effect of lipoxin A(4) (LXA(4)) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA(4) at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca(2+)](i) of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca(2+)](i) of monocytes in the PE group were significantly higher than those in the control group. LXA(4) significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA(4), in the PE group, the [Ca(2+)](i) concentration of monocytes was significantly reduced. It was concluded that LXA(4) may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.

It analyzed the definition, mechanism, characteristics of Mirror Visual Feedback and summarized the application of mirror visual feedback in recovering upper limb function after stroke patients at home and abroad, so as to provide evidences for the further research in China.

Objective To investigate protective effects and mechanisms of lipoxinA4(LXA4)on human umbilical vein endothelial cells(HUVEC)under hypoxia in vitro.Methods The HUVEC culture were divided into groups as followed:added M199 cudture medium as normal contml groups,added CoCl2 to mimic hypoxia in vitro as hypoxia group and added different concentrations of LXA4(1,10,100 nmol/L)were added to the induced hypoxiai HUVEC as agents intervention group.Morphological changes of HUVEC were observed by using inverted phase contrast mieroscope.The influence of LXA4 on cell survival was investigated by methyl thiazolyl tetrazolium(MTT) assaying method after the treatment with different concentrations of LXA4 and 100 nmol/L lipoxinA4 according to different time(4,8,12 and 24 hours).The expression of von-willebrand factor (vWF) was detected by immunocytoehemistry method. The changes of cytosolic Ca2+ were measured by laser scanning confocal microscope. Results ( 1 ) Morphological changes:the cells under hypoxia lost its normal shapes and showed necrosis, while the cells cocuhured with 100 nmol/L LXA4 were normal appropriately. (2)Survival rate: the survival rates of HUVEC under hypoxia was (40. 1±3.9) % and increased to ( 52. 9 ± 1.4) %, (64. 1 ± 3. 3 ) %, ( 76. 6 ± 1.6) % respectively when added with LXA4 with concentration of 1,10, 100 nmol/L into culture medium. There was significant different survival rate when compared with that of hypoxia group. (3) The level of vWF: The expression of vWF was decreased with the increasing concentrations of LXA4 added into culture medium, the gray values were 203.9 ±0. 7 in 1 nmol/L,204.6 ±0. 9 in 10 nmoL/L,191.8 ±0. 5 in 100 nmol/L respectively, which reached statistical difference in comparison with that of hypoxia groups (P<0. 05). (4) Confocai analysis:the intracellular free Ca2+ concentrations of HUVEC were intensified with LXA4 treatment. Conclusions LXA4 plays an important role in keeping the normal shape of HUVEC under hypoxia, can enhance survival of hypoxial HUVEC and decrease the level of vWF in cytoplasm. The protective mechanism might be via decreasing mitochondria Ca2+ overload and increasing cytoplasm Ca3+ by nucleus Ca2+ transference.