Objective To investigate the effect of immunoglobulin G (IgG) in bullous pempbigoid (BP) blister fluid on the secretion of chemokines by human keratinocytes. Methods IgG was obtained from the blister fluid of patients with bullous pemphigoid and sera of normal human controls, then purified by sequential precipitation with caprylic acid and ammonium sulfate. The immunological activity of blister fluid was tested before and after the purification by BP180 ELISA kit. Keratinocytes were isolated from the foreskin tissue of yong adults, and subjected to primary culture. After 3 passages, the primary keratinocytes were harvested and subcultured in the presence of purified IgG of 0.5, 1, 2, 4 and 8 g/L, respectively, for 24 hours, or IgG of 4 g/L for 3, 6, 12, 24 hours, respectively, followed by the detection of levels of eotaxin, monocyte chemoattractant protein (MCP)-1 and interleukin (IL)-8 in the supemate of keratinocytes by ELISA. Results The valence of IgG remained unchanged after the purification with caprylic acid and ammonium sulfate. Compared with IgG from the sera of normal controls, that from bullous pemphigoid blister fluid sig- nificantly enhanced the secretion of IL-8 by keratinocytes in a time- and dose-dependent manner (both P < 0.01 ). Neither eotaxin nor MCP-1 was detected in the supemate of control IgG-treated, BP IgG-treated or untreated keratinocytes. Condusions The IgG in BP blister fluid has been proved to stimulate the secretion of IL-8 by cultured human keratinocytes, which may be involved in the pathogenesis of BP.

Objective To study the corelation of mucosal involvement and corticosteroid dosage in the control of bullous pemphigoid(BP). Methods One hundred and three in-patients with bullous pemphigoid hospitalized during 1988 - 2002 were retrospectively analyzed for their mean corticosteroid dosage for controlling the disease and the duration for complete relieving the skin lesions, as well as the duration to start corticosteroid tapering. All the patients were divided into two groups: group A (37 cases) with mucosal involvement and group B (66 cases) without mucosal involvement. Results The mean corticosteroid dosage used to control the disease in group A was much higher than that in group B (t = 3.488, P 0.05). Conclusion Patients with mucosal involvement require a higher dosage of corticosteroids to control the disease.

Objective To investigate the role of genistein (Gen) in the expression of connective tissue growth factor (CTGF) induced by parathyroid hormone (PTH) in human renal tubular epithelia cells. Methods Real-time PCR, Western blotting and reporter gene assay were employed to detect the role of Gen in PTH-induced CTGF expression in HK-2 cells. The activity of NF-κB was measured by EMSA to investigate the mechanism by which PTH induced CTGF expression in HK-2 cells. Inhibitors of NF-κB signaling pathway were used to ascertain which signal pathway was involved. Results HK-2 cells had basic amount of CTGF mRNA and protein, which, however, increased significantly after treatment with PTH, and the luciferase activity increased to a higher level as compared with control group after treatment with 10-10 mol/L PTH for 12 h (1.89±0.08 vs 0.99±0.03, P＜0.01). Gen decreased the expressions of CTGF mRNA and protein induced by PTH in dose-dependent manner. The NF-κB of nucleus was inactivation without PTH, while the activity of NF-κB significantly increased after exposed to PTH, with the maximal response of PTH at a concentration of 10-10 mol/L and the best stimulating time at 30 minute. The NF-κB inhibitor PDTC reduced the increase of CTGF transcript levels in response to PTH stimulation. Gen blunted PTH-mediated NF-κB activation. Conclusion Gen inhibits CTGF expression induced by PTH through bloking NF-κB signaling pathway in human renal tubular epithelial cells.

Objective To evaluate the effect of remifentanil on Toll-like receptor 2 (TLR2) mRNA expression during renal ischemia-reperfusion (Ⅰ/R) in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (group S),Ⅰ/R group and remifentanil group (group R).Renal Ⅰ/R injury was produced by clamping the bilateral renal arteries for 45 min followed by reperfusion in Ⅰ/R and R groups.Bilateral renal arteries were only exposed but not clamped in group S.Remifentanil 1.0 μg · kg-1 · min-1 was infused via the tail vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead in S and Ⅰ/R groups.The animals were sacrificed at 15 min before ischemia and 6 and 24 h of reperfusion,and the renal specimens were obtained for examination of the pathological changes (with light microscope) and for determination of the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by ELISA) and expression of TLR2 mRNA (by RT-PCR) and cell apoptosis (by double staining and flow eytometry).The apoptotic rate was calculated.Results Compared with group S,TLR2 mRNA expression was significantly up-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were increased at 6 and 24 h of reperfusion in Ⅰ/R and R groups.Compared with group Ⅰ/R,TLR2 mRNA expression was significantly down-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were decreased at 6 and 24 h of reperfusion in group R.The pathological changes were significantly attenuated in group R as compared with group Ⅰ/R.Conclusion The mechanism by which remifentanil reduces renal Ⅰ/R injury is related to down-regulation of TLR2 expression and decrease in TLR2 activity and inhibition of inflammatory responses in renal tissues and cell apoptosis in rats.

Objective To investigate the role of fluid therapy in treatment of septic shock in children.Methods A total of 48 pediatric patients with septic shock were enrolled.Twenty-seven patients were treated with volume expansion and 21 patients were treated with fluid resuscitation.There were no differences in use of antibiotics,inotropic and vasoactive agent in two groups.The time needed for haemodynamic stability,occurrence of pulmonary edema.length of PICU stay,and mortality were compared between two groups.Results Compared with volume expansion group,the time needed for haemodynamic stability significantly decreased in fluid resuscitation group ((216.10± 168.13) minutes vs (121.63 ± 75.59) minutes) (P < 0.05). There were no significant difference in occurrence of pulmonary edema between volume expansion group and fluid resuscitation group (14.8% vs 9.5% . P > 0.05) .Compared with volume expansion group,length of PICU stay significantly decreased in fluid resuscitation group ((6.188 ±3.250) days vs (3.944±2.711) days,P < 0.05).Compared with volume expansion group,mortality significantly decreased in fluid resuscitation group (40.7% vs 14.3%,P < 0.05) . Conclusions Rapid fluid resuscitation was associated with early reach of haemodynamic stability,short stay in PICU,improved survival rate and no increase in the risk of complication in pediatric patient with septic shock.

Background The progenesis and development of dry eye is closely associated with the stability of tear-film.Dry eyes often show the visual disturbance.Therefore,to evaluate the stability of tear-film based on visual quality and explore the relevant dynamic vision change in dry eyes is of important significance.Objective The aim of this study was to evaluate the dynamic changes of objective scattering index (OSI),modulation transfer function cut off (MTF cut off),objective visual acuity (OVA) with tear-film in the patients with mild or moderate dry eye disease using Optical Quality Analysis System.Methods A clinical diagnosis test was performed.Forty-two eyes of 42 consecutive patients with mild or moderate dry eyes were included in Tianjin Eye Hospital from December 2013 to May 2014 based on ‘ The Clinical Diagnosis and Treatment Recommendations for Dry Eye from Consensus of Expert Advice in 2013’,and 29 normal eyes of age-and gender-matched 29 healthy volunteers were simultaneously included as the healthy control group.The dynamic retinal images under non-blinking condition during 15 seconds were recorded by OQAS-Ⅱ system,and the OSI,MTF cut off and OVA values at 0,5,10 and 15 seconds were obtained,and the differences of OSI,MTF cut off and OVA between 0 second and 15 seconds were calculated (△OSI,△MTF cut off,△OVA).The dynamic changes of △OSI,△MTF cut off and △OVA with time were evaluated.The sensitivity and specificity of △OSI,△MTF cut off and △OVA for the differential diagnosis of dry eyes were analyzed by receiver operating characteristic curve (ROC curve).Results The OSI values at 5,10 and 15 seconds in the mild or moderate dry eye group were significantly higher than those at 0 second or those in the healthy control group (all at P＜0.05),and the MTF cut off values and OVA values at 5,10 and 15 seconds in the mild or moderate dry eye group were significantly lower than those at 0 second or those in the healthy control group (all at P＜0.05).The △OSI,△MTF cut off and △OVA values were 1.03±1.53,(-8.37±9.64)c/d and-0.30±0.32 in the mild or moderate dry eye group,which were significantly higher than 0.06± 0.16,(-0.37 ± 5.27) c/d and-0.03 ± 0.18 in the healthy control group (t=4.10,-5.19,-5.28,all at P＜0.05).The area under ROC curve was 0.90,0.79 and 0.78 in △OSI,△MTF cut off and △OVA,respectively.The sensitivity and specificity of △OSI was 78.6％ and 93.1％ respectively at the diagnostic cut off of 0.3.Conclusions The optical quality parameters related with tear-film stability are gradually reduced with extending of time in mild or moderate dry eyes,but they are stable in healthy eyes.The detecting outcomes by OQAS-Ⅱ system can reflect the dynamic status of tear-film,and this method is of important clinical value for the differential diagnosis and optical quality evaluation in dry eyes.

Objective To explore the bias between the real pressure and the measured values when handheld pressure gauge (HPG) was used to monitor intermittently the pressure in the intubation balloon,so as to provide some measures for the correct use of HPG.Methods In the first part of the study,HPG was used to measure the pressure with the balloon connected with a three-way tube with which to control the inflation and deflation in a laboratory to measure the pressure in the air bag.After gaining the deviation in this in vitro experiment,it was tested and verified in vivo in adult patients undergoing endotracheal intubation.Results After 132 times of measurements,it was found that measurement with a HPG might result in an inherent loss (3.928 ± 0.291) cmH2O (1 cmH2O=0.098 kPa,t =155.273,P =0.000) between inflation value [(30.000 ± 0.000) cmH2O] and measured value [(26.072 ± 0.291) cmH2O].In addition,after 214 times repeated measurements,the pressure loss during disconnection of the gauge was as high as (1.196 ± 0.954) cmH2O (t=18.348,P=0.000) between filled values [(30.000 ± 0.000) cmH2O] and measured values [(28.804 ± 0.954) cmH2O] and it was named as error loss.At last,the total error was verified by clinical test,and it was (5.270 ± 2.583) cmH2O (t=29.632,P=0.000) between pressure of filled value [(30.000 ± 0.000) cmH2O] and measured value [(24.730 ± 2.583) cmH2O].Conclusions When the balloon pressure was Monitored intermittently with HPG,the real value should be the measured value plus the error.In addition,subglottic aspiration should be done before the connection of the balloon to the gauge to prevent the secretions on the cuff falls into the deeper airway,and to maintain the cuff pressure at 30 cmH2O.

BACKGROUND: The existence of minimal residual leukemia cells is the main cause for the recurrence of acute leukemia in children, and immunological biological therapy has attracted more and more attentions in the various methods from eliminating minimal residual disease. Previous studies have found that interferon α-2b can effectively inhibit the increase of tumor cells in vivo in children with neuroblastoma and malignant lymphoma, whether it can inhibit the increase of leukemia cells?OBJECTIVE: To investigate the effects of interferon α-2b in vitro on leukemia cells.DESIGN: A comparative observation taking human promyelocytic leukemia HL-60 cell line as the material.SETTING: Cell Culture Room; Immunological Laboratory; Cell Room, Institute of Pediatrics, Affiliated Hospital,Medical College of Qingdao University.MATERIALS: HL-60 cell line was provided by Shandong Institute of Basic Medical Sciences. Interferon α-2b was purchased from Megagene Company Fluorescein isothiocyanate (FTTC) rabbit-anti-rat Ig solution (CatEK001) and CD13 anti-human monoclonal antibody solution (Cat. DK013Y) were purchased from Union Stem Cell & Gene Engineering Co.,Ltd.METHODS: The experiments were carried out in the Institute of Pediatrics, Affiliated Hospital, Medical College of Qingdao University from March to September 2005. HL-60 cells culture system was established in vitro, and the oncentration was adjusted to 1×109 L-1. The cells were divided into control group and experimental group. In the experimental group, each well was added by interferon-α-2b with the terminal concentration of 5×105, 1×106, 2×106,5×106 and 1×107 U/L, respectively. In the control group, each well was added by saline of the same volume. The cells were cultured continuously for 48 hours. The morphological changes of HL-60 cells were observed using Wright's staining under light microscope; Cell apoptosis was observed using acridine orange/ethidium bromide double staining; Antigen expression and maturation and differentiation on cell membrane were observed by determining CD13 protein expression; Proliferation and activity of HL-60 were detected with methyl-thiazol-tetrazolium (MTT) assay.MAIN OUTCOME MEASURES: The occurrence of apoptosis was judged according to the uniformity and staining of HL-60 nuclear chromatin; HL-60 cell proliferation was judged according to the absorbance (A) value; The maturation of HL-60 cells was judged according to the number of positive CD13 cells.RESULTS: ① HL-60 cell apoptosis: The cells were cultured for 48 hours. When the concentration of interferon α-2b was 5×105 U/L, there were mainly early apoptotic HL-60 cells; When the concentration was 1×107 U/L, there were mainly late apoptotic cells, and the apoptotic rate was significantly higher than those in the control group (P ＜ 0.01 ).② HL-60 cell proliferation: The A values in the experimental groups treated with interferon α-2b of 2×106 U/L and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01). ③ Maturation of HL-60 cells: The percentages of positive CD13 cells in the experimental groups treated with interferon α-2b of 1 ×106 and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01).CONCLUSION: It is concluded that interferon α-2b can enhance the apoptosis, inhibit the proliferation and promote maturation and differentiation of HL-60 cells.

Objective To evaluate the effect of preconditioning with nimodipine on postoperative cognitive dys function of aged rats.Methods Ninety healthy male Sprague-Dawley rats,aged 18 months,weighing 400-500 g,were randomly divided into 3 groups (n =30 each) using a random number table:nimodipine control group (group N),surgery group (group S),and nimodipine + surgery group (N+ S group).In N and N + S groups,nimodipine 1 mg/kg was intraperitoneally injected,while the equal volume of normal saline was given instead in S group.30 min later,group N inhaled pure oxygen for 2 h,and S and N + S groups inhaled 1.8 ％ isoflourane for 2 h when splenectomy was performed.Morris water maze test was performed on 1 day before operation and 1st,3rd and 7th days after operation.After the end of Morris water maze test at 1 day before operation and 1st and 7th days after operation,10 rats were sacrificed and brains were removed and hippocampi were isolated for determination of apoptosis in hippocampal neurons,intracellular [Ca2+] i in cytoplasm,and hippocampal Bcl-2 and Bax mRNA expression and for examination of ultrastructure of hippocampal neurons.Results Compared with the value before administration,the escape latency was significantly prolonged,the frequency of crossing the original platform was decreased,apoptotic rate and [Ca2+]i were increased,Bcl-2 mRNA expression was down-regulated,and Bax mRNA expression and Bax/Bcl-2 mRNA ratio were up-regulated at each time point after operation in S and N + S groups,and no significant changes were found in the parameters mentioned above in N group.Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was inecreased,apoptotic rate and [Ca2+]i were decreased,Bcl-2 mRNA expression was up-regulated,and Bax mRNA expression was down-regulated at each time point after operation in group N + S.Pathological changes were found in S and N + S groups and the damage was severer in S group than in N + S group.Conclusion Nimodepine preconditioning can prevent postoperative cognitive dysfunction of aged rats,and inhibition of calcium overloadinduced apoptosis in hippocampal neurons may be involved in the mechanism.

Objective To investigate the changes of follicular helper T cells (Tfh cells) and Tfh cells associated molecules in the peripheral blood (PB) of patients with malignant lymphoid diseases (MLD) dynamically, and explore their roles on pathogenesis of the diseases. Methods Fifty-five patients with MLD were enrolled in this study,including 9 patients with acute lymphocyte leukemia (ALL), 30 patients with non-Hodgkin lymophoma (NHL) and 16 patients with multiple myeloma (MM), and 10 healthy controls (NC) of similar age were also enrolled. The percentage of CD4+CXCR5+cells (Tfh cells) and expression of ICOS+, PD1+among the T cells were detected by flow cytometry (FCM), while the levels of interleukin 21 (IL-21) in plasma were detected by ELISA tests. Results The percentage of Tfh cells and expression of ICOS and/or PD-1 in PB of all untreated patients were significantly higher than those of NC (all P< 0.01), and MM group< ALL group 0.05), and apparently lower than those who achieved PR (P< 0.05). The percentages of Tfh cells in patients with T-ALL and T-NHL were higher than those in patients with B-ALL and B-NHL (P> 0.05), and much higher than NC (P< 0.01). The concentration of IL-21 in patients were much higher than that in NC [(326.56±32.44) pg/ml] (P<0.01), and MM group