Objective To explore the clinical significance of immunocyte subsets before and after immunosuppressive therapy in the peripheral blood of patients with immune thrombocytopenic purpura (ITP).MethodsThe percentages of immunocyte subsets in the peripheral blood of 35 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD56+,CD19+ lymphocytes and CD4+/CD8+.Results The percentages of CD3+ T lymphocyte (61.58 ± 6.45 ) %,CD4+ T lymphocyte (28.38 ±4.89)% and the ratio of CD4+/CD8+ 0.99 0.22 in patients with ITP were lower than those in healthy controls[( 67.85 ± 4.68 ) %,( 38.00 ± 3.37 ) %,1.54 ± 0.13,all P ＜ 0.05].After immunosuppressive therapy,the percentages of CD3+ T lymphocyte ( 69.41 ± 5.03 ) %,CD4+ T lymphocyte (38.17 ±3.18)% and the ratio of CD4+/CD8+ 1.60 ±0.15 recovered to control levels.The percentages of CD8+ T lymphocyte (29.20 ±4.50)% and CD19+B lymphocyte ( 17.74 ±4.14)% were higher than those in healthy controls[( 24.82 ± 2.93 ) % and ( 12.09 ± 3.51 ) %,all P ＜ 0.05].After the immunosuppressive therapy,the percentages of CD8+ T lymphocyte ( 24.06 ± 3.02 ) % and CD19+ B lymphocyte ( 10.90 ± 3.55 ) %recovered to control levels.There were no significant difference of the percentage of CD56+ lymphocyte among ITP patients ( 15.80 ± 2.85 )%,ITP patients after immunosuppressive therapy ( 15.16 ± 2.77 )% and healthy controls ( 16.36 ± 2.75 ) %.ConclusionThe aberrant immunocyte subsets are involved in the pathogenesis of ITP,and detection of immunocyte subsets might be helpful for the diagnosis and determination of therapeutic outcome of ITP.

OBJECTIVE To evaluate the possibility that CdCl2 induces autophagy and apoptosis in HEK293 cells,and the role of extracellular regulated protein kinases (ERK1/2) and AKT proteins in autophagy. METHODS Green fluorescence protein(GFP)-light chain 3B(LC3B)expression plasmid was transfected into HEK293 cells. After 24 h,HEK293 cells were induced with CdCl2 2,4,8 and 10μmol·L-1 for 12 h. The expression of GFP-LC3B was detected by fluorescent microscopy. HEK293 cells were induced with CdCl2 2,4,8 and 10μmol · L-1 without transfection of GFP-LC3B for 12 h while autophagic vacuoles were observed by transmission electron microscopy. The expression of LC3B-Ⅱ/Ⅰproteins and the phosphorylation levels of ERK1/2 and AKT were analyzed by Western blotting. Apoptosis was detected by flow cytometry microscopy. HEK293 cells were treated with 3-MA 20μmol · L-1+CdCl2 10 μmol · L-1 for 12 h before cleaved caspase 3 protein was detected by Western blotting. RESULTS When HEK293 cells were exposed to CdCl2(≤10μmol · L-1)for 12 h,cytoplasmic GFP-LC3B punctuates were observed under the fluorescence microscope,and autophagic vacuoles were observed under an electron microscope. The expression of LC3B-Ⅱ/Ⅰ,p-ERK1/2 and p-AKT proteins was significantly increased in CdCl2-induced cells(P<0.05,P<0.01). Moreover,apoptosis was observed. The addition of 3-MA 20μmol · L-1+CdCl2 10μmol · L-1 enhanced apoptosis. Cleaved capase 3 protein expression was significantly increased(P<0.01). CONCLUSION CdCl2(≤10μmol·L-1)can induce autophagy in HEK293 cells. ERK1/2 and AKT proteins might be associated with the activation of autophagy that is accompanied by apoptosis,suggesting that autophagy can inhibit apoptosis at certain concentrations of CdCl2.

Insulin-like growth factor-1 receptor (IGF-1 R) widely exists in the surface of various types of cells and is closely associated with the formation and development of tumor cells.It also provides a new direction for the targeted therapy for tumors.This article reviews the expression,development,and progression of IGF-1R in pancreatic cancer and research advances in IGF-1R as a target for tumor treatment.

Objective To detect the doubling time of airway smooth muscle cells of human and rat by the xCELLigence in-strument,a real time cellular analyzer.Methods The airway smooth muscle cells were separated by collagenase-pancreatin digestion from the rat airway.Then,added to the different holes of E-plate of xCELLigence instrument with the planting destiny of 3 000 cells/well.and so were the human airway smooth muscle cells.The E-plate was then placed on the xCELLi-gence instrument to monitor cell proliferation for 100 hours to calculate the doubling time by using the RTCA Software Package 2.0 software.Results The doubling time of human airway smooth muscle cell calculated by the real time cellular analyzer was 23.96±0.47 h,which was consistent with the data provided by the reference.The doubling time of rat airway smooth muscle cell was 18.62±0.15 h,and the computational process was simple,time-saving and also effective.Conclusion The xCELLigence instrument can be used to calculate doubling time of airway smooth muscle cells of human and rat, which provides experimental methods and reference data for the basic respiratory disease research.

Background and purpose:Radiofrequency ablation (RFA) was one of the best combined treatments for hepatocarcinoma. CT was commonly used to evaluate treatment response and recurrent disease. However it was diffi cult to evaluate treatment effectiveness of RFA and to detect recurrent nodule especially the size below 1cm. Digital subtraction arteriography(DSA) may provide fi nal diagnosis of recurrent nodule after RFA for hepatocarcinoma. Our purpose was to analyze the signs of DSA after RFA for hepatocarcinoma in order to provide possible reference for treatment effectiveness and the imaging follow-up methods. Methods:17 patients with primary liver cancer (n=15) or hepatic metastasis(n=2) were enrolled in this study. Common hepatic arteriography or super-selective angiography of suspicious tumor area were performed on all patients. Results:On DSA, most ablated regions presented as round or ovoid low density areas with no stain. At the peripheral zone of the lesion, fi ve signs after RFA could be found: Stain of localized granulation tissue, arterio-portal fi stula, hemorrhage, recurrence and no abnormal fi ndings. Local or intrahepatic recurrence occurred in 9 cases. Conclusion:Hepatic DSA has great value in detecting local recurrence of hepatic tumor after RFA treatment. DSA is superior to CT in detecting marginal or intrahepatic recurrent nodule below 1 cm.

Objective To observe the efficacy of cumulative dose of polyethylene glycol-interferon (Peg-IFN)α-2a combined with ribavirin in patients with decompensated hepatitis C virus (HCV)-related liver cirrhosis , and to evaluate the effects of anti-virus therapy on the progress of the disease . Methods From January 2005 to March 2009 ,patients with decompensated HCV-related liver cirrhosis were enrolled ,also included patients received partial splenic embolization .Peg-IFNα-2a combined with ribavirin therapy was given to patients whose blood cell met interferon (IFN) therapy standards .The dosage of Peg-IFNα-2a and ribavirin was adjusted according to the tolerance of the patients .After the treatment ,the patients were followed-up for 24 weeks .The patients whose blood cell did not meet IFN therapy standards and the patients unwilling to receive anti-virus therapy were assigned to control group and were followed-up for 96 weeks .The total amount of medication was calculated according to cumulative exposure dose . Sustained virological response (SVR ) , recurrence rate , liver function and disease progression were observed .The t test or Chi-square test was performed for comparison between groups and rate of disease progression was analyzed with Kaplan Meier curve .Results After anti-virus therapy , SVRs of patients with cumulative dose of Peg-IFNα-2a and ribavirin over 60% (include 60% ) were 27 .3%(12/44) and 27 .7% (13/47) ,respectively ;the recurrence rates were 7/19 and 35 .0% (7/20) ,respectively . In patients with cumulative dose less than 60% ,SVRs were 1/7 and 1/4 ,respectively ,and the recurrence rates were both 1/2 ;the differences of different doses was not statistically significant (all P>0 .05) .At the 24th week of follow-up after therapy ,the Child-Pugh score of combined therapy group was 7 .9 ± 1 .4 , which was lower than that before treatment (8 .5 ± 1 .2) ,and the difference was statistically significant (t=2 .33 ,P=0 .02) .At the 96th week of follow-up ,the Child-Pugh score of control group was 10 .0 ± 1 .6 ,which was higher than that before treatment (8 .5 ± 1 .4) ,and the difference was statistically significant (t=5 .82 , P<0 .01) .The disease progression rate of combined therapy group was 15 .7% , which was lower than that of control group (32 .4% ) ,and the difference was statistically significant (χ2=4 .34 ,P= 0 .04) .Conclusion The application of non-standard dosage of Peg-IFNα-2a combined with ribavirin in the patients with decompensated HCV-related liver cirrhosis can achieve virological response once the cumulative dose reached certain standards ,improve Child-Pugh scores of patients and slow disease progression .

Background The biomeasurement and imageology of retinal nerve fiber layer(RNFL) thickness showed the damage of retinal structure in the early and middle stage of glaucomatous eye,however,the subtle functional damage of glaucoma can not be timely reflected only with automatic static perimeter.Microperimetry is a method of quantitatively assessing mean sensitivity (MS) of macular zone.Objective This study was to evaluate and compare the macular functional change in early and middle stage of primary open angle glaucoma (POAG) and chronic angle-closure glaucoma (CACG) with MP-1 microperimeter.Metbods This trail protocol was approved by Ethic Committee of Peiking University Third Hospital,and written informed consent was obtained from each subject prior to entering the study group.A cross-sectional and case-controlled study was designed.A total of 126 eyes from 126 subjects were enrolled in the trail,including 53 eyes of 53 normal subjects,50 eyes of POAG patients and 23 eyes of CACG patients.A macular 10° program was set with MP-1 microperimetry to record the MS of various subareas.The macula was zoned into central 2°,6°,and 10° visual fields and 4 quadrants in each ring.The MS of different rings and subareas was detected and compared among POAG patients,CACG patients and normal controls.Results The MS values of central 2°,6°,10° and whole macular area were (15.09 ± 3.03),(15.72 ± 3.22),(13.99 ± 3.63) and (14.91±3.07)dB in the POAG group,which were significantly lower than those of (17.29±1.59),(18.05±1.24),(16.76±1.89) and (17.37±1.46)dB in the normal control group (all at P=0.000).The MS values of central 2°,6°,10° and whole macular area was (16.00±2.39),(15.83±2.63),(14.45±3.15) and (15.42±2.54) dB in the CACG group,and the reduced MSs were seen at the 6°,10° rings and whole macular area in the CACG group compared with the normal control group (P =0.004,0.013,0.011).Within the 6° ring,the MS values in the inferotemporal quadrant were declined in the POAG group and CACG group compared with the normal control group (P =0.000,0.022),but the difference was not statistically significant between the POAG group and the CACG group (P =0.311).In addition,the MS value in the inferonasal quadrant was significantly lower than that of the normal control group (P =0.005); while that in the CACG group was not significantly different in comparison with the normal control group (P=0.119).In the POAG group,the MS value of the inferotemporal quadrant was significantly lower than that of the superonasal or superotemopral quadrant (P =0.043,0.016),but no significant differences were found among the 4 quadrants in the CACG group (all at P＞0.05).Conclusions The mild damage of retinal function appears in the early and middle stage of POAG and CACG.More serious MS reducing occurs in the inferotemporal and inferonasal quadrants of POAG.

Objective To evaluate the effect of propofol on local field potential of prefrontal cortex in rats in order to investigate the reason why pmpefol leads to cognitive dysfunction.Methods Thirty healthy male SD rats weighing 190-230 g were randomly divided into 3 groups(n=10 each):intralipid group(group C),low dose propofol group(group P1)and high dose propofol group(group P2).In group C,P1 and P2,10% intralipid 0.01 ml·kg-1·min-1,pmpofol 0.1 mg·kg-1·min-1,and propofol 0.5 mg·kg-1·min-1 were infused iv through the caudal vein for 2h respectively.The modified Morris water-maze (MWM) test was performed twice a day for 5 consecutive days one day after administration.The escape latency,swimming time in platform quadrant,percentage of swimming distance in platform quadrant in the total swimming distance and the fLrst central point were recorded.Propofol 0.1 and 0.5 mg·kg-1·min-1 were infused iv in group P1 and P2 respectively 14 days after propfol administration.Local field potential of prefrontal cortex was recorded at 1-2 h of administration.Results Compared with group C and P1,the escape latency was prolonged,the swimming time in platform quadrant was shortened,and the percentage of swimming distance in platform quadrant in the total swimming distance and the first central point were signifieandy decreased in group P2(P<0.05).The complexity and power spectrum were significantly lower in group P2 than in group P1(P<0.05).Conclusion The high dose of propofol Can inhibit prefrontal cortex neuronal discharge activity to result in cognitive dysfunction.

Objective To investigate the effects of grape seed proanthocyanidin(GSPE) on mitochondrial injury in hippocampus and learning-memory impairment after obstructive sleep apnea hypoxia in rats.Methods Male SD rats(n=80) were randomly divided into control group,model group,low dose of GSPE treatment group and high dose of GSPE treatment group.Rats in control group were exposed in air,the model group were suffered from intermittent hypoxia conditions (50 ml/L,8-hour-intermittent hypoxia everyday,and the duration of experiment 2 and 6 weeks,respectively).Mitochondrion pathology in hippocampal region was observed using electron microscope;malondialdehyde (MDA) contents and superoxide dismutase activity were detected by colorimetry and apoptotic cells was measured by TUNEL method.The cognitive function of rats in each group was assessed with the Morris water maze (MWM).Results After hypoxia,mitochondrion was significantly injured.The MDA contents were increased(79.86 ± 2.52,88.26 ± 2.86) and SOD level decreased (70.67 ± 6.70,64.26 ± 7.86).The number of neural apoptotic cells was significantly enhanced (9.68 ± 0.79,15.9 ± 2.92).MWM test showed that the escaping latency was prolonged and the frequency of crossing the platform was decreased (P ＜ 0.05).Compared with that in the model group,low dose of GSPE decreased MDA contents (76.38 ± 1.96,82.16 ±2.02),increased SOD level(76.20 ± 6.86,70.58 ± 6.86),and decreased apoptotic cells (6.60 ± 0.69,9.54 ±1.36).MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased in GSPE treatment groups(P ＜ 0.05).Compared with low dose of GSPE,high dose of GSPE decreased MDA contents increased SOD level and decreased apoptotic cells.MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased (P＜ 0.05).Conclusion GSPE can attenuate mitochondrial injury and improve learning-memory function after obstructive sleep apnea hypoxia.

Objective To explore the clinical significance of a series of cytokines and peripheral blood immunocyte subsets before and after immunosuppressive therapy in patients with immune thrombocytopenia (ITP).Methods The percentages of immunocyte subsets in the peripheral blood of 20 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD4+/CD8+,CD1~.ELISA was applied to detect the level of serum TNFα,IL-2,IL-6,IL-4,IL-10,IL-11,IL-17,IL-27,transforming growth factor β (TGFβ),thrombopoietin (TPO) of 20 patients with ITP and 20 healthy controls.Results The percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+ / CD8+ T lymphocyte in patients with ITP were lower than those in healthy controls [(62.66 ± 6.58) ％ vs (69.93 ± 4.81) ％,(29.46 ± 5.02) ％ vs (39.08 ± 3.50) ％,0.97 ± 0.35 vs 1.56 ± 0.26,all P ＜ 0.05].After immunosuppressive therapy,the percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+/CD8+ T lymphocyte [(71.49 ±5.16)％,(39.25 ±3.21)％ and 1.56 ±0.28] recovered to the same levels in healthy controls.The percentage of CD8+ T lymphocyte and CD19+ B lymphocyte in patients with ITP were higher than those in the healthy controls [(30.28 ±4.63)％ vs (25.90±3.06)％,(18.92 ± 4.27)％ vs (13.17 ± 3.64)％,all P ＜ 0.05].After treatment of immunosuppressive therapy,the percentage of CD8+ T lymphocyte and CD19+ B lymphocyte [(25.16 ± 3.45) ％ and (11.98 ± 3.68) ％] recovered to the similar levels in healthy controls.The serum levels of IL-4,IL-6,IL-11,IL-17 and TPO in patients with ITP were significantly higher than those in healthy controls.While TGFβ level was significantly decreased.There was no significant difference of IL-27 between ITP patients and healthy controls.After the treatment of immunosuppressive therapy,IL-4,IL-6,IL-11,IL-17,TPO and TGFβ were down-regulated while IL-27 was up-regulated.There was no significant difference of IFNγ,TNFα,IL-2 and IL-10 among ITP patients before or after immunosuppressive therapy and healthy controls.Conclusions The present study suggests that the aberrant immunocyte subsets and cytokines are involved in the pathogenesis of ITP.Hyper-function of Th2 and Th17,dysfunction of Treg cells,up-regulation of IL-27,IL-11,TPO and other factors may contribute to the pathogenesis of ITP.