Magnetic manipulation of particles/cells in microfluidic chips is a promising research field.This paper stated the operation mechanism and the main means of manipulation, including separation, concentration, capture, arrangement and assembly.Especially, the concept of particles/cells separation was emphasized with different criteria, like sizes, shapes, and magnetic properties.In addition, the effects of the channel geometry, the intensity and distribution of the magnetic field, and the types of magnetic liquid (paramagnetic salt solutions and ferrofluids) on the performance of the magnetic manipulation were also compared.The prospective to the prospect of magnetic manipulation about particles/cells in microfluidic chip was also depicted.

Objective To investigate the relationship between endothelial damage and p120-catenin (p120-ctn) in a model of paraquat intoxication,and the modulatory effect of mangiferin on p120-ctn.Methods Human umbilical vein endothelial cells (HUVECs) were cultured in two compartment spreading apparatus in vitro.The endothelial cells were divided into three groups:control group (cultured in DMEM with 10％ fetal bovine serum),paraquat group (paraquat was added to the medium with final concentration of 0.05 μmol/L) and mangiferin group (cultured in medium with addition of paraquat for 30 minutes,then mangiferin was added in a final concentration of 20 μmol/L).The cellular permeability at 6,12,24,48,72 hours after culture in the three groups was measured.The expressions of p120-ctn 1A,p120-ctn 3A mRNA and p120-ctn protein were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot analysis.The distribution of p120-ctn protein was observed by immunofluorescence.Results Compared with control group,cellular permeability in paraquat and mangiferin groups were increased with prolongation of time,and peaked at 72 hours [(29.86 ± 3.98)％,(24.39 ± 2.79)％ vs.(11.71 ± 1.67)％,both P＜0.05].The cellular permeability was significantly lower in mangiferin group than that in paraquat group at different time points (all P＜0.05).At 6 hours after intoxication,the expressions of p120-ctn 1A,p 120-ctn 3A mRNA (gray value) and p 120-ctn protein (gray value) were significantly lower in paraquat group than those in control group (p120-ctn 1A mRNA:0.150 ± 0.024 vs.0.433 ± 0.024,p120-ctn 3A mRNA:0.316 ± 0.043 vs.0.701 ±0.020,p120-ctn protein:0.485 ±0.031 vs.0.763 ±0.038,all P＜0.01).The expressions of p120-ctn 1A,p120-ctn 3A mRNA and p120-ctn protein were significantly higher in mangiferin group than those in paraquat group from 6 hours on (p120-ctn 1A mRNA:0.281 ± 0.021 vs.0.150 ± 0.024,p120-ctn 3A mRNA:0.602 ± 0.042 vs.0.316 ± 0.043,p120-ctn protein:0.675 ± 0.031 vs.0.485 ± 0.031,all P＜0.01),and they were gradually increased with prolongation of time,and peaked at 72 hours (p120-ctn 1A mRNA:1.376 ±0.128 vs.0.150 ± 0.024,p120-ctn 3A mRNA:1.251 ± 0.059 vs.0.316 ± 0.043,p120-ctn protein:0.844 ± 0.050 vs.0.485 ± 0.031,all P＜ 0.01).Under upright fluorescence microscope,p120-ctn was mainly distributed in the cell membrane in control group,with a slight expression in cytoplasm,and no expression in the nuclei.With prolongation of time,p120-ctn expression in the cell membrane was gradually decreased in paraquat group,while it was increased in the cytoplasm and nuclei,with blurring of cell membrane and widening of cellular gap.p120-ctn expression was improved on the cell membrane in mangiferin group at corresponding time points,with decreased in expression in nuclei and cytoplasm.Conclusion The p120-ctn protein plays an important role in the enhancement of endothelial permeability in paraquat intoxication,and mangiferin may attenuate endothelial injury in paraquat intoxication possibly through modulation of p 120-ctn protein.

Objective To investigate the expressions of Yes-associated protein-1 (YAP1) in gallbladder mucosal epithelium of normal persons,in patients with simple/calculous cholecystitis,and in patients with gallbladder carcinoma;and to study the mechanism of YAP1 in gallbladder carcinoma development.Methods Immunohistochemistry was used to detect the expression and distribution of YAP1 protein in 50 persons with normal gallbladder,101 patients with simple cholecystitis/calculous cholecystitis and 100 patients with gallbladder carcinoma.RT-PCR and western-blot were used to detect the mRNA and protein levels of YAP1 in normal and malignant gallbladder mucosal epithelium cells.siRNA was used to shut down the expression of YAP1 in SGC996 cells.MTT was used to test cell vitality.Flow cytometry was used to measure cell cycle.Results Immunohistochemistry revealed the expression rates of YAP1 in the gallbladder carcinoma group,the cholecystitis/gallstone group and the control group to be 87.0％ (87/100),56.4％ (57/101) and 5.0％ (1/20),respectively (P ＜ 0.01).The YAP1 protein levels were higher in gallbladder carcinoma tissues and cells when compared to normal tissues and cells.RT-PCR showed the mRNA levels of gallbladder carcinoma cells to be 12.5 ± 1.2 times of normal gallbladder mucosal epithelial cells (P ＜ 0.05).After using siRNA to shut down the YAP1 expression,EMT associated proteins were down-regulated,cell vitality was decreased,and cell cycle was arrested in the S-phase.Conclusions YAP1 is closely related to cell proliferation and metastasis of gallbladder carcinoma.It may promote tumor progression through epithelial-mesenchymal transition.transition;Tumor progress

Animals ; Hypothermia ; blood ; Hypoxia ; blood ; Malondialdehyde ; blood ; Myocardial Ischemia ; blood ; Rabbits ; Superoxide Dismutase ; blood

BACKGROUND:Stem cell transplantation provides a new approach to treat chronic renal disease.Specific marking and in vivo tracing of stern cells are the basis of studies in this field.However,the marking methods appropriate for all cells remain uncertain.OBJECTIVE:To observe the in vivo location and differentiation of 4',6-diamidino-2-phenylindole (DAPI) and green fluorescence protein (GFP)-Iabeled cells in adriamycin nephrosis rats so as to explore an efficient labeling and tracing method for metanephric mesenchymal cells (MMCs) derived from embryonic rats.DESIGN,TIME AND SETTING:Grouping comparative observation was performed at the Second Xiangya Hospital of Central South University from April to December 2007.MATERIALS:A total of 60 female SD rats,weighing 180-220 g,of dean grade,were used to establish models of adriamycin nephrosis.METHODS:DAPI and MMCs infected with GFP and DAPI were respectively injected into addamycin nephrosis via the tail vein.DAPI and GFP distribution in the frozen sections was detected at 1,3,and 5 weeks,postoperatively.In addition,GFP expression in renal tissues was detected by ABC immunoenzymatic staining method.MAIN OUTCOME MEASURES:DAPI and GFP-labeled Cell grafts in adriamycin nephrosis rats were compared.The changes of GFP-transfected MMCs at different time points were observed.RESULTS:DAPI positive cells were observed in tubular structures after 1 weeks of injection of DAPI-labeled cells and DAPI alone,and remained existing at 5 weeks,but the florescence was reduced with time.GFP-transfected MMCs were able to survive and integrate into tubular structures after 1 week,and remained existing at 5 weeks.Moreover,the fluorescence was not reduced.ABC immunoanzymatic staining showed that only a few GFP-positive MMCs appeared in glomerular tufts,and mainly distributed in cytoplasm.Semi-quantitative evaluation of GFP show that the positive cell rate in rats with early application was greater than that with advanced application,and the positive rate was increased with time.CONCLUSION:Liposome mediated GFP gene transfer was an efficient labeling in vitro and suitable tracing method for cell differentiation experiment in vivo,suitable for short-term tracing and observation of transplanted cells.

Objective To choose zebrafish as the experimental animal model for studying the spatiotemporal expression rule of rap1 gen in zebrafish embryo early development process .Methods The Rap1 gene fragment was cloned from the zebrafish emby‐oscDNA ,then the Rap1 gene fragment and pCS2+ plasmid were performed the in vitro connetion and recombination was extracted , the combinant plasmid was correct after the double enzyme digestion ,colony PCR and sequencing identification .T3 RNA polymer‐ase in vitro transcription system was used to obtain the digoxin (DIG)‐labeled anti‐sense mRNA probe of Rap1 gene .The whole mount in situ hybridization method was adopted to detect the Rap1 expression in zebrafish embryo early development process . Results The positive hybridization signal of Rap1 gene was detected at the cell division junction region of 0 .75 hpf ,animal pole of 3 .70 hpf and 6 .00 hpf ,and notochord of 12 .00 -72 .00 hpf .Conclusion Rap1 gene might be involved in the early development process of notochord nervous system in zebrafish .

Objecfive To detect the functional repair of metanephric mesenchymal cells (MMCs) transplantation in adriamycin (ADR)-induced glomerulopathy rats. Methods A total of 90 Sprague-Dawley female rats were randomly divided into three groups:ADR group (n=40,rats were injected via the tail vein with O.25 mg ADR/100 g body weight on days 1 and 21),ADR- MMCs group(n=40,rats were injected via the tail vein with 5×10~6-7×10~6 MMCs 8 weeks after the second ADR administration),control(n=10).All the rats were scarified 8 weeks after MMCsinjection.Pathology and collagen IV expression in renal tissue were examined.Moreover,matrix metalloproteinases 2 (MMP-2) and matrix metallopmteinases 9 (MMP-9) expression in the renal tissue were also detected with immunohistochemistry,and quantity analysis of protein and gene was further demonstrated with Westem blot and RT-PCR analysis,respectively. Results There were no significant differences in tubulointerstitial injury score and glomerulosclerosis degree between ADR group and ADR-MMCs group(P>0.05).Compared with ADR group,collagen Ⅳ and MMP-2 expression decreased, MMP-9 expression incrased in renal tissue of ADR-MMCs group, and the difference was significant (P<0.05). Conclusion MMCs transplantation may have potentially therapeutic effect on renal tissue fibrosis of adriamyein-induced glomerulopathy in rats, and the signaling pathways of MMPs appear to be involved in these processes.

Objective To investigate cyclic mechanical stimulation on expression of connective tissue growth factor (CTGF) in osteoblast-like cells (MG63) and to explore the rote of MAPK involved in the process.Methods Expressions of CTGF protein and mRNA in MG63 cells were detected by Western blot and RT-PCR,respectively. Phosphorylation levels of p38, ERK, JNK were examined by Western blot. Results Cyclic mechanical stimulation upregulated expressions of CTGF protein and mRNA. The levels reached a maximal response of 2-3 fold after 3-6 h. ERK and JNK signal pathways were activated by cyclic mechanical stimulation, the phosphorylated proteins increased within 10 min of stretch, phosphorylated ERK reached maximal levels by 60 min of stretch, phosphorylated JNK reached maximal levels by 15-30 min of stretch, but not for p38 signal pathway.Only the inhibitior of JNK signal pathway (SP600125) markedly suppressed stretch-induced CTGF expression,meanwhile the inhibitors of ERK (PD98059) and p38 (SB203580) did not show such effect. Conclusion Cyclic mechanical stimulation upregulates CTGF expression via JNK-dependent pathway in MG63 cells.

To explore the effects of recombined Neuregulin on the heart of healthy Macaca mulatta, 10 healthy adult Macaca mulatta were randomly divided into two groups and were injected with the same doses of recombined Neuregulin and normal saline, respectively. At the same time, related indices were detected by 2-dimensional echocardiography and M-mode echocardiography. All indices were compared between the two groups and among different phases. Recombined Neuregulin had effects on LVEDD (left ventricular end-diastolic diameter), LVEDV (left ventricular end-diastolic volume), LVESV (left ventricular end-systolic volume) and SV (Stroke volume), and the effects changed with time. However, no significant changes were seen on EF (Ejection fraction) and FS (Fractional shortening). In conclusion, recombined Neuregulin has effects on the left ventricular volume of healthy Macaca mulatta, but no significant effect on cardiac contractility.
Animals ; Echocardiography ; methods ; Female ; Heart Ventricles ; anatomy & histology ; diagnostic imaging ; drug effects ; Macaca mulatta ; Male ; Myocardial Contraction ; drug effects ; Neuregulins ; biosynthesis ; genetics ; pharmacology ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Stroke Volume ; drug effects

Acute Disease ; Glucocorticoids ; Graft Rejection ; Heart Rate ; Humans ; Hyperglycemic Hyperosmolar Nonketotic Coma ; Liver Transplantation