Objective To explore the roles of urokinase type PA (uPA) and uPA receptor (uPA R) in the course of repair after human embryo corneal alkali burn in vitro . Methods After the alkali burn model in vitro was established successfully, some techniques such as the observation of cell culture and morphology, ICC and image analysis were used. Results No significant difference was found between the cells from corneal limbus and central cornea and almost all of them expressed AE1/AE3. The expressions of uPA and uPA R increased to the maximum at about 24 h after alkali burn. uPA expression distributed mainly in the cytoplasm but uPA R expression distributed mainly in the cell membrane. Conclusion The corneal epithelial cells of comparatively high purity can be acquired by tissue culture. There probably exists difference in development and vitality between embryo and adult cornea. There might be commonness of the roles of uPA and uPA R in epithelial tissue.

OBJECTIVETo study the influence of bone marrow mesenchymal stem cells (MSCs) transplantation on hypoxic pulmonary hypertension (HPH) in rats.

METHODSSD rats MSCs were separated, cultivated, identified and labeled by the green fluorescence protein (GFP) gene virus and transplanted in vitro. Healthy male SD rats were randomly divided into four groups: Normal control group (NC group) and HPH group (eight rats respectively), HPH+ MSCs transplantation group and HPH+ VEGF+ MSCs transplantation group (twenty-four respectively). The test employed atmospheric intermittent low oxygen method to establish the rat model of pulmonary hypertension and stem cells were transferred and transplanted. The rats' mean pulmonary artery pressure (mPAP) was observed; right ventricular hypertrophy index (RVHI) was calculated; the morphological change of lung small artery in various groups of rats was observed under the microscope; the distribution of lung small artery and adenovirus transfection fluorescently labeled MSCs was observed under a fluorescent microscope after 7, 14 and 28 days when stem cell was transplanted.

RESULTSFor NC group, the mPAP (mmHg) was 15.5 +/- 1.5 after twenty-eight days while the mPAPs for HPH , MSCs and MSCs+ VEGF were 26.1 +/- 1.9, 21.6 +/- 2.7 and 20.1 +/- 2.9 respectively which were apparently higher than that of NC group (P < 0.01) and compared with HPH group (P < 0.01), which declined clearly. There was no significant difference between MSCs and MSCs+ VEGF. After twenty-eight days, RVHI for NC group was 0.28 +/- 0.02 while the RVHI for HPH, MSCs and MSCs + VEGF were 0.43 +/- 0.07, 0.34 +/- 0.03 and 0.35 +/- 0.01 respectively which was apparently higher than that of NC group (P < 0.01) but which was clearly lower than that of MSCs and MSCs+ VEGF (P < 0.05) and there was no significant difference between MSCs and MSCs + VEGF. For HPH group, pulmonary arteriole wall became apparently thicker, the lumen became significantly narrow and nearly obstructed after twenty-eight days, the endothelial cells were incomplete; compared with HPH group, pulmonary arteriole wall of MSCs group became thin, the lumen was smooth and the completeness of endothelial cells was improved. Whereas for MSCs and MSCs + VEGF, these changes were not significantly clear.

CONCLUSIONAfter MSCs transplantation, mPAP and RVHI decline sharply and lung small artery remodeling is improved which partially reverses HPH process; there is no significant difference between VEGF together with MSCs transplantation group and pure MSCs.

Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; etiology ; metabolism ; surgery ; Hypoxia ; complications ; Male ; Mesenchymal Stem Cell Transplantation ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; pharmacology

Aged ; Aged, 80 and over ; Complement C3 ; metabolism ; Humans ; Immunoglobulin A ; blood ; Male ; Silicosis ; blood ; Tumor Necrosis Factor-alpha ; blood

Objective To investigate the effects of the intravenous immunoglobulin (IVIG) on the expression of FcγR Ⅱ b and Toll-like receptor 4 (TLR4) in children with Kawasaki disease (KD).Methods 25 children with KD and 15 healthy controls were enrolled in this study.The levels of TLR4 expression on monocytes were assessed by flow cytometry.Real-time PCR was performed to detect the transcription levels of FcγR Ⅱ b,TLR4 signaling molecules(MyD88,TRAF-6,TAK1) and cytokines(IL-1β,IL-6,TNF-α) in monocytes.The concentrations of TNF-α in plasma was determined by enzyme-linked immunosorbent assay (ELISA).Results (1) Compared with healthy controls,the expression of TLR4 and the signaling molecules in the patients were significantly up-regulated but decreased after treatment with IVIG (P＜0.05).(2)The levels of FcγR Ⅱb expression from the patients were higher than those from healthy controls,and decreased after treatment with IVIG(P＜0.05).(3) The levels of cytokine factor expression such as IL-1β,IL-6 and TNF-α in the patients were found to be higher than those in healthy controls (P＜0.05) ; plasma concentrations of TNF-α were significantly elevated during acute KD (P＜0.05).(4)There were significantly correlations to follow between FcγR Ⅱb and the levels of cytokine,TLR4 expression in monocytes (P ＜0.05).Conclusion IVIG can up regulate FcγR Ⅱb expression and down regulate TLR4 expression which might inhibit the inflammatory response.

Objective To explore the relationship of monocyte chemoattractant protein-1(MCP-1)and oxygen-induced retinopathy(OIR).Methods Thirty-six newborn SD rats were divided into 2 expanded litters,18 of which were exposed to 50 mL/L oxygen and then 10 mL/L oxygen in alternating 24-hour periods(experiment group),an additional 18 rats as control rats were raised simultaneously in room air(control group).On postnatal 14 days(P14),oxygen-exposed rats were removed to room air.The eyeballs of 6 rats from each group were enucleated and fixed by formaldehyde at postnatal 14 days(P14),postnatal 17 days(P17),and postnatal 21 days(P21)respectively,and then cross sectioned.The nuclei of proliferative retinal vessels were counted through the crosssections to measure the average retinal capillary density index(RCDI)when stained with hematoxylin and eosin(HE)under light;the expression of MCP-1 was measured by immunohistochemistry.Results There existed obviously differencc between 2 groups in the field of both RCDI and the expression of MCP-1 at the same time point [t(P14)=6.69 P=0.001,t(P17)=3.43 P=0.006,t(P21)=2.37 P=0.039;t(P14)=40.45,t(P17)=43.44,t(P21)=17.45 Pa=0],when RCDI and the expression of MCP-1 were compared among the different time points with in the same group,there existed obviously difference among three time points in the experiment group(F=17.74 P=0.0001;F=421.5 P=0),but no differencc in control group(F=0.016 P=0.984;F=0.006 P=0.994).There existed positive correlation between the expression of MCP-1 and the value of RCDI in experiment group(r=0.822 P=0).Conclusions Neovascularization resulting from OIR occurs before room air recove-ry.MCP-1 is upregulated and subsequently downregulated in OIR.Neovascularization in the OIR model appear to be associated with increased retinal MCP-1.

25% or decrease by 15%,at termination ofTCI propofol and 5 rain after termination for determination of arterial plasma propofol concentrations which werecompared with the values predicted by the TCI system.Performance was determined by the median performanceerror(MDPE),the mean absolute performance error(MDAPE),the divergence and the wobble.Results A totalof 200 blood samples were taken.The mean value for MDPE was 14.9%(-21.6%-42.9%),for MDAPE23.3%(6.9%-62.5%),for divergence -1.9%?h~(-1)(-32.7%?h~(-1)-23.0%?h~(-1))and for wobble 18.9%(4.2%-59.6%).PE did not increase with time but with increasing target propofol concentration.ConclusionThe anesthesia was satisfactory in all patients.The predictive performance of Diprifusor TCI system is consideredacceptable for clinical purposes.The wobble is relatively big,for Chinese patients the pharmacokinetic model needsimproving.

Objective To investigate the changes and roles of Fc gamma receptors (FcγR) expressed on monocytes in the immune pathogenesis of Henoch-Schonlein purpura(HSP).Methods Thirty children of HSP and 15 health controls were enrolled in this study.The expressions of FcγR Ⅰ and FcγRⅢ on monocytes were determined by flow cytometry,and real-time PCR was performed to detect the transcription levels of FcγR Ⅱ a,FcγR Ⅱ b,cytokines(IL-1 β,IL-6,TNF-α,IFN-α),chemokine(IP-10,RANTES,iNOS),and BLyS/April in monocytes isolated by microbeads.The plasma concentrations of IL-4,IL-10 and TNF-α were analyzed by enzyme-linked immunosorbent assay (ELISA).Results (1) The expressions of FcγR Ⅰ and FcγR Ⅲ on monocytes in patients with HSP were significantly up-regulated compared with healthy controls.Transcription level of FcγR Ⅱ a on monocytes in patients with acute HSP was found to be higher than that in healthy controls while the inhibitory FcγR Ⅱ] b mRNA was significant down-regulated(P＜0.05),which resulted in a higher ratio of FcR Ⅱ a/Ⅱ b in patients with acute HSP.(2) The expressions of cytokine/chemokines factor such as IL-1 β,IL-6,TNF-α,IP-10,RANTES,and iNOS in patients with HSP was detected to be higher than those in healthy controls(P＜0.05).In addition,expression levels of BLyS/April were up-regulated during acute HSP(P＜0.05),the positive correlations were observed between the FcγR Ⅱ a/FcγR Ⅱ b and the cytokine/chemokines factor in monocytes(P＜0.05).(3) Plasma concentrations of IL-4,IL-10 and TNF-α were significantly elevated during acute HSP(P＜0.05),and a negative correlation was observed between concentrations of TNF-α and the mRNA level of FcγR Ⅱb in monocytes.Conclusion The abnormal expression of the cytokines and the imbalance of stimulatory and inhibitory FcγR of monocyte in acute HSP.

Objective:To explore the psychological and behavioral pattern of state speech anxiety of participants of college speech contest under two conditions.Methods:Narrow band measurements of both Subjective Uncomfortable Degree(SUD)and objective behavioral assessments were used under two conditions.Results:Under lab condition,SUD rat-ings differed among four time bands,while the behavioral ratings failed to show any difference.Under field condition,for the prepared theme speech,SUD ratings differed among four time bands while they failed to show any difference for the improvisational speech.For both speech forms,several behavioral items displayed a pattern of habituation.Conclusion:Under both conditions,the psychological and behavioral pattern of participants' state speech anxiety display a pattern of habituation while the behavioral pattern is affected more by different conditions.

Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.