Objective: To observe the effects of angiopoietin-like 4 (ANGPTL4) gene on the growth of different hepatocellular carcinoma (HCC) cell lines. Methods: The expression of ANGPTL4 was tested by reverse transcriptase-polymerase chain reaction and Western blotting in 9 HCC cell lines and immortal liver cells. Huh-7 cells and MHCC-LM3 cells with low ANGPTL4 expression and MHCC-97L cells with moderate ANGPTL4 expression were transfected with fused ANGPTL4-GFP and empty pEGFP-N1 plasmids via Lipofetamine 2000, respectively. GFP-positive cells were screened with G418 and sorted by flow cytometry (FCM) to establish stable cell lines with over-expression of ANGPTL4, Cell array was prepared and Ki-67 fluorescent immunocytochemical techniques were used to detect in vitro proliferation of the three HCC cell lines. Xenografted tumor formation in nude mice was used to observe the effect of ANGPTL4 on the growth of the three HCC cells in vivo. Results: ANGPTL4 mRNA had low expression in Huh-7 and MHCC-LM3 cell lines, moderate expression in MHCC-97L cells, and high expression in Hep3B cells. The results of cell array showed that cell proliferation rate was lower in Huh-7 cells (P = 0.000 74), but higher in MHCC-LM3 (P = 0.073 08) and MHCC-97L (P = 0.011 52) cells with over-expression of ANGPTL4 compared with the cells transfected with empty plasmid. Over-expression of ANGPTL4 significantly inhibited xenografted tumor formation of Huh-7 cells (P = 0.001 10), but markedly promoted the xenografted tumor formation of MHCC-LM3 (P=0.057 50) and MHCC-97L (P = 0.018 91) cells in nude mice. Conclusion: ANGPTL4 plays different roles in the growth of different HCC cell lines, but the effects of ANGPTL4 on the growth of the same cell line was consistent in vitro and in vivo.

By reshaping the cornea without the creation of a stromal flap, excimer laser corneal surface ablation eliminates flap-related complications and avoids the risk of ectasia that may occur after laser assisted in situ keratomileusis ( LASIK ) . Post-operative pain is one of the most significant disadvantages of surface ablation and thus the management of pain and discomfort following surface ablation is of great importance. We summarize mechanism of corneal pain and current approaches to pain management after surface ablation.

0. 05) and the specificity is higher than that of the SEA-ELISA (P

ObjectiveTo investigate the effect and underling mechanism of water-soluble CO-releasing molecules (CORM-3)on the alleviation of allograft rejectionafter mouse kidney transplantation.Methods A mice kidney transplantation model was established using C.FVB-Tg (Itgax-DTR/GFP)57Lan/J or C57BL/6J (H-2Kb) mice as donors,and Balb/c (H-2Kd) mice as recipients.After donor nephrectomy,kidney was preserved in UW solution which contained CORM-3 or iCORM (inactive CO-releasing molecules) for 24 h in 4℃.Recipient survival after removal of both na? ve kidneys,serum creatinine as well as graft histology was observed.In the C.FVB-Tg(ItgaxDTR/GFP) 57Lan/J donors,rDCs were acquired in vitro and selected by magnetic cell sorting (MACS) after graft nephrectomy.The expression of activation markers,CD80 and CD86,on rDC was assessed by using flow cytometry.ResultsThe graft medium survival time was 40.5 days in the iCORM group and 70 days in the CORM-3 group respectively (P＜0.05).CORM-3 preserved the graft function as shown by significantly lower serum creatinine (P＜0.05; or P＜0.01) and alleviated graft pathology injury.Diffuse infiltration of mononuclear cells in the interstitial tissues,moderate tubulitis and partial glomerular sclerosis were found in the iCORM graft kidney,while the CORM-3 graft kidney displayed almost normal histology.Meanwhile,CORM-3 suppressed the expression of CD80 and CD86 in donor-derived rDC.ConclusionCORM-3 can alleviate allograft rejection,prolong the graft survival,and improve kidney function in mouse kidney transplantation,probably via inhibiting rDC activation.

Objective To evaluate the effect of the serum of rats with hepato-pulmonary syndrome (HPS) on the expression of caveolin-1 and VE-cadherin in pulmonary microvascular endothelial cells (PMVECs).Methods Among the 40 healthy Sprague-Dawley rats,aged 3-4 months,weighing 220-250 g,20 rats were taken randomly for establishment the model of HPS which was produced by chronic ligation of the common bile duct,and the left 20 rats served as sham operation group.Primary PMVECs were harvested from healthy adult Sprague-Dawley rats and inoculated in ECM culture medium or on 96-well culture plate.The PMVECs of 4th-9th generation were randomly divided into 2 groups (n =36 each):control group (group C) and HPS group.In group C,the serum obtained from normal rats in sham operation group was added to PMVECs,while the serum obtained from rats with HPS was added in HPS group.The final concentration of serum was 10％.After being incubated for 12,24 and 36 h (T1-3),the expression of caveolin-1 and VE-cadherin in PMVECs was detected by Western blot,and the PMVEC adhesion rate and proliferation were determined by CKK-8 method.Results Compared with group C,the expression of caveolin-1 and VE-cadherin was significantly down-regulated,the cell adhesion rate was decreased,and the proliferation of PMVECs was enhanced in HPS group.Conclusion The serum of rats with HPS induces weakened PMVEC contact inhibition through down-regulating caveolin-1 and VE-cadherin expression.

AIM: To explore the effect of pyrrolidine dithiocarbamate (PDTC), an NF-κB inhibitor, on the proliferation and apoptosis of human multiple myeloma U266 cells and its mechanisms.METHODS: The U266 cells were treated with PDTC at different concentrations (0, 25, 50, 100 and 200 μmol/L) in vitro.The growth inhibitory rate of the U266 cells was detected by CCK-8 assay and cell counting.The cell cycle of the U266 cells was determined by flow cyto-metry, and the apoptosis was examined by flow cytometry with Annexin V-FITC/PI staining.The effect of PDTC on the expression of DNA methyltransferase 1 (DNMT1) at mRNA and protein levels was measured by RT-qPCR and Western blot, respectively.The effects of PDTC on the protein levels of NF-κB (P65), DNMT1, Bcl-2, cyclin D1, cleaved caspase-3 and cleaved caspase-8 were determined by Western blot.RESULTS: The protein level of NF-κB (P65) was decreased after treatment with PDTC for 48 h or 72 h.PDTC inhibited the proliferation of U266 cells in both dose-and time-dependent manners.After treatment with PDTC for 48 h, the percentage of U266 cells in G2 phase increased compared with control group (P<0.05).PDTC induced the apoptosis of U266 cells in a dose-dependent manner.The expression of DNMT1 at mRNA and protein levels decreased (P<0.05).The results of Western blot showed that the expression of Bcl-2 in PDTC groups decreased, while the protein levels of cyclin D1, cleaved caspase-3 and cleaved caspase-8 were higher than those in control group (P<0.05).CONCLUSION: The NF-κB inhibitor PDTC inhibits the proliferation of U266 cells by inducing cell apoptosis.It may be related to the down-regulated expression of DNMT1, cell cycle arrest and activation of the apoptotic pathways.

To investigate whether human umbilical cord plasma (HUP) can be used to culture human cord blood mesenchymal stem cells (HUCMSCs), we collected 20 surplus HUP. After being treated with salting out and diasysis, the HUP were used to culture HUCMSCs as 10% volume, and compared with fetal bovine serum (FBS). Morphological characteristics, growth curve and reproductive activity of HUCMSCs cells were observed. The concentration of bFGF and noggin secreted by HUCMSCs cultured with HUP and FBS medium were detected by ELISA. It was found that compared to FBS, the morphology, reproductive activity and characteristic of HUCMSCs cell cultured with HUP were not distinctively different from FBS. The concentration of bFGF in HUP group was significantly higher than that of FBS group, and the concentration of noggin was also different in the two groups. So we concluded that HUP could be used to culture HUCMSCs for a long-time, and the HUP mediumcoild could be more suitable for the culture of human embryonic stem cell (hESC).
Animals ; Cattle ; Cell Culture Techniques ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Plasma ; chemistry ; Serum ; chemistry

Objective To evaluate the effect of compound tropicamide eye drops on the measurement of central corneal thickness and to investi-gate the duration of the effect.Methods In this trial, 240 eyes of 120 myopic patients undergoing corneal laser refractive surgery were selected, and central corneal thicknesses were measured before treatment and 1 , 4 h after administration of compound tropicamide eye drops using OrbscanⅡ anterior segment analysis system and SP -2000P non -contact specular microscope, respectively.Results The central corneal thick-ness was (545 ±27),(559 ±31) and (544 ±26) μm before and 1 and 4 h after administration, measured with Orbscan Ⅱ anterior segment analysis system, whereas being ( 508 ± 26 ) , ( 521 ±29 ) and (506 ±24) μm measured with SP -2000P non -contact specular microscope.The central corneal thickness measured with both OrbscanⅡsystem and SP-2000 P microscope increased 1 h after administration of compound tropicamide eye drops as compared to that before administra-tion ( P <0.01) , and the central corneal thickness measured with both Orbscan Ⅱ system and SP -2000P microscope decreased 4 h after administration of compound tropicamide eye drops as compared to that 1 h after administration ( P <0.01 ) , while no significant difference was observed in central corneal thickness measured with either Orbscan Ⅱ system or SP -2000 P microscope between 4 h after administration of Mydrin eye drops and before application. Conclusion Central corneal thickness increases 1 h after the application of compound tropicamide eye drops, and the effect of compound tropicamide eye drops on central corneal thickness is eliminated at 4 h following administration. During the examination prior to excimer laser corneal refractive surgery, measurement of central corneal thickness should be performed before or 4 h after administration of compound tropicamide eye drops.

Aged ; Female ; Femoral Neck Fractures ; diagnostic imaging ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Radiography ; Retrospective Studies ; Treatment Outcome