Objective To study the effects of Treponema pallidum membrane protein Tpp17 on ac-tivation of human umbilical vein endothelial cells (HUVECs) in vitro and to understand its role in the immu-nopathogenesis of syphilis .Methods HUVECs were co-cultured with recombinant protein Tpp 17.Then the expressions of TNF-α, MCP-1, ICAM-1 and E-selectin at mRNA and protein levels in supernatants were re-spectively detected by enzyme-linked immunosobent assay ( ELISA ) and fluorescent real-time quantitative PCR.The adhesive ability of THP-1 cells was observed by fluorescence microscopy after co-cultured pretrea-ted HUVECs with recombinant protein Tpp 17 with Calcein-AM labeled THP-1.Pretreated HUVECs were cultured in the lower chamber of Transwell with recombinant protein Tpp 17 and monocytic THP-1 cells were cultured in the upper chamber of Transwell .After that, the migration of monocytic THP-1 cells was evalua-ted by using fluorescence microscopy .Results Compared with the blank control group , the expression of TNF-α, ICAM-1, E-selectin, especially the MCP-1, were enhanced by recombinant protein Tpp 17 of Trepo-nema pallidum.Moreover, Tpp17 improved the adhesive ability and migration of monocytic THP-1 cells, es-pecially the latter.Conclusion Treponema pallidum membrane protein Tpp17 might play a certain role in the immunopathogenesis of syphilis by enhancing the expression of TNF-α, MCP-1, ICAM-1 and E-selectin, and by promoting the adherence ability and migration of monocytic THP-1 cells.

Objective To investigate the in vitro effects of Treponema pallidum membrane protein Tpp17 on the permeability of endothelial barrier for further investigation on the immunopathogenesis of syphi-lis.Methods A cellular model of in vitro monolayer was established by using human umbilical vein endo-thelial cells ( HUVECs) .Cell-ELISA and a TMB kit were respectively used to measure the expression of VE-cadherin and the flux of horseradish peroxidase ( HRP) by monolayer HUVECs after stimulation with the re-combinant Tpp17 (rTpp17) protein.THP-1 cells stained with Calcein AM were added to the top of HUVEC monolayer in Transwell culture.Then, the numbers of THP-1 cells in the upper wells and beneath the HUVEC monolayer were counted by using a fluorescence microscope.The rTpp17 protein-treated HUVECs were fixed in 4%buffered paraformaldehyde and stained with rhodamine-phalloidin for observing the distri-bution of F-actin under a confocal laser scanning microscope.Results Compared with the control group, the expression of VE-cadherin in HUVECs was decreased, while the permeability of HUVEC monolayer was increased upon the stimulation with rTpp17 protein (P<0.05).Moreover, rTpp17 protein-induced F-actin redistribution and increased transendothelial migration of THP-1 cells were observed in rTpp17 protein-trea-ted HUVECs as compared with those of the control group (P<0.05).Conclusion Treponema pallidum membrane protein Tpp17 could suppress the expression of VE-cadherin and enhance the redistribution of F-actin, resulting in an enhanced transendothelial migration of THP-1 cells and an increased permeability of HUVEC monolayer.The Tpp17 protein might play an important role in the immunopathogenesis of syphilis.

Objective To investigate mechanisms underlying the regulation of the permeability of vascular endothelial cells by the Treponema pallidum membrane protein Tpp47.Methods Human umbilical vascular endothelial cell (HUVEC) monolayers were established as a model,and were directly cultured with the presence of recombinant Tpp47 protein (rTpp47-treated group),or boiled and inactivated rTpp47 (negative control group).Some HUVEC monolayers,which were pretreated with the RhoA/ROCK signal pathway inhibitor Y-27632 for 30 minutes and then cultured with the presence of rTpp47,served as the pretreatment group.After 1-and 4-hour additional culture,enzymelinked immunosorbent assay (ELISA) was performed to estimate the permeability of these cell monolayers to horseradish peroxidase (HRP).After 12 hours of culture,rhodamine-phalloidin was used to stain cytoskeletal proteins,and confocal laser scanning microscopy was performed to observe the arrangement of the cytoskeletal protein F-actin.Western-blot analysis was conducted to measure the expressions of RhoA in HUVECs treated with rTpp47 or inactivated rTpp47.Results The supernatant level of HRP (expressed as the absorbance value at 450 nm) was significantly higher in the rTpp47-treated group than in the negative control group (0.81 ± 0.10 vs.0.39 ± 0.09,P ＜ 0.05),but no significant difference was observed between the pretreatment group (0.51 ± 0.10) and rTpp47-treated group or negative control group (both P ＞ 0.05) after 1-hour culture.Similarly,the rTpp47-treated group showed significantly increased levels of HRP compared with the pretreatment group and negative control group (2.31-± 0.14 vs.1.21 ± 0.12 and 0.73 ± 0.12,both P ＜ 0.05),while there was no significant difference between the pretreatment group and negative control group after 4-hour culture.The expression of RhoA in HUVECs treated with rTpp47 was significantly higher than that in those treated with inactivated rTpp47.Confocal laser scanning microscopy showed that rTpp47 treatment led to the rearrangement of F-actin in HUVECs followed by the formation of stress fibers in cytoplasm,while Y-27632 could partly inhibit the rearrangement of F-actin.Conclusion The recombinant Treponema pallidum membrane protein Tpp47 can regulate the permeability of vascular endothelial cells through the RhoA/ROCK signal pathway.

Objective To evaluate the effect of Treponema pallidum membrane protein Tpp47 on vascular endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs) were classified into multiple groups to be cultured with various concentrations (50,100,200,400 and 800 μg/L) of the recombinant protein Tpp47 or lipopolysaccharide (LPS) for different durations (3,6,12,24 and 48 hours).Then,enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin in the culture supernatant of,fluorescence-based real-time quantitative PCR to quantify the mRNA expressions of ICAM-1 and E-selectin in,HUVECs.The 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay was used to evaluate the proliferation activity of HUVECs treated with Tpp47 (400 μg/L) and LPS (200 μg/L) respectively for 24 hours.To estimate the effect on adhesion ability,some HUVECs were pretreated with Tpp47 (400 μg/L) and LPS (200 μg/L) respectively for 24 hours followed by coculture with THP-1 human monocytic leukaemia cells for 6 hours,then,the adhesion of HUVECs to THP-1 cells was visualized by fluorescence microscopy.The cells receiving no treatment served as the blank control.Results A significant increase was observed in the supernatant level (expressed as the absorbance value at 450 nm) of ICAM-1 for HUVECs treated with Tpp47 of 400 μg/L for 24 hours (1.28 ± 0.03 vs.0.90 ± 0.01,t =18.28,P ＜ 0.05) and that of E-selectin for HUVECs treated with Tpp47 of 400 μg/L for 12 hours (0.51 ± 0.01 vs.0.13 ± 0.03,t =18.19,P＜ 0.05) compared with untreated HUVECs.The adhesion rate to THP-1 cells was significantly higher in HUVECs pretreated with Tpp47 for 24 hours than in untreated HUVECs (56.1％ ± 1.9％ vs.16.3％ ± 2.1％,x2 =12.65,P ＜ 0.05).The cell proliferation rate was 19.5％ ± 1.7％ in HUVECs treated with Tpp47,significantly higher than that in untreated HUVECs (10.0％ ± 3.1％,x2 =3.92,P＜ 0.05),but lower than that in those treated with LPS (41.2％ ± 3.7％,x2 =10.42,P ＜ 0.05).Conclusions The recombinant membrane protein Tpp47 could enhance HUVECs to proliferate and adhere to monocytic THP-1 cells,suggesting a certain role of Tpp47 in the pathogenesis of syphilis.

Objective To investigate the expression of hepatocyte growth factor (HGF)and vascular endothelial growth factor (VEGF)in acute myeloid leukemia(AML) and its significance in the progress of AML.Methods Serum levels of HGF and VEGF in AML patients,including 25 de novo cases,17 cases with complete remission( CR),16 cases without remission and 12 cases with relapse,were measured by enzyme-linked immunosorbent assay( ELISA ).The data were compared with those of normal controls.Results The serum levels of HGF were( 1357.29 ± 358.64)ng/L in de novo patients,(1175.93 ± 306.71 )ng/L in patients without remission and( 1261.21 ± 340.83 )ng/L in patients with relapse,which were significantly higher than those in normal control(232.62 ± 99.13 ) ng/L and CR patients ( 256.65 ± 94.32 ) ng/L( F =78.35,P ＜ 0.01 ).The serum levels of VEGF were ( 253.84 ± 49.14 ) ng/L in de novo patients,( 245.87 ± 54.68 ) ng/L in patients without remission and(264.75 ± 62.52) ng/L in patients with relapse,which were significantly higher than those in normal control ( 97.61 ± 16.19 ) ng/L and CR patients ( 99.76 ± 15.93 ) ng/L ( F =68.65,P ＜ 0.01 ).Compared with de novo group,the serum levels of HGF and VEGF in CR patients decreased significantly but showed no significant difference from those in normal controls(P ＞0.05).The expression of HGF and VEGF in acute myeloid leukemia was positively correlated with each other ( r =0.49,P ＜ 0.05 ).Conclusion The expression of HGF and VEGF was closely related with the incidence and development of acute myeloid leukemia.Preventing HGF expression may be a possible strategy for leukemia therapy in the future.

ObjectiveTo observe the clinical efficacy of vesiculation moxibustion plus Levofloxacin in treating chronic bronchitis.MethodEighty-three eligible patients with chronic bronchitis were randomized into an observation group of 43 cases and a control group of 40 cases. The control group was intervened by orally taking Levofloxacin,while the observation group was additionally intervened by vesiculation moxibustion. The clinical efficacy in the two groups and the safety of vesiculation moxibustion were evaluated.ResultThere was a significant difference in comparing the total effective rate between the two groups (P<0.05), and the observation group was superior to the control group; vesiculation didn’t cause fever, obvious swelling, or big blisters, etc.ConclusionVesiculation moxibustion plus Levofloxacin can produce a satisfactory clinical efficacy in treating chronic bronchitis, and it’s safe and reliable without obvious adverse reactions and worth application in clinic.

Objective To investigate the clinical value and sonographic features of locally advanced cervical carcinoma using transvaginal color Doppler ultrasound(TVCDU). Methods A retrospctive analysis was carried on 2-D sonograms. And color Doppler flow imaging(CDFI) in 106 cases, confirmed by pathology and surgery. Results In locally advanced cervical carcinoma, the characteristerics on TVCDU were cervical enlargment, cervical masses accompanying with uteral effusion and extracervical infiltrative masses.The rich cervical blood flow signals demonstrated as called “fireball" sign. In early cervical carcinoma,the cervix showed no apparent cervical enlargment. The blood flow signals were increased.The average RI was 0.41. Conclusions TVCDU which is very valuable in judging the infiltration extent and in differential diagnosis preoperatively can be used as a routine examination method before locally advanced cervical carcinoma treatment.

Objective To assess the effects of captopril on renal perfusion of diabetic mellitus(DM)rabbits at early stage by contrast-enhanced ultrasound (CEUS).Methods Of 58 rabbits,6 were control group,37 were made as DM model successfully by alloxan and were randomly divided into 4 groups:untreatment group (n =16) and treatment group A,B and C (n =7 averagely).The treatment groups were given captopril(25 mg/kg weight) everyday by intragastric administration after the DM model established and renal pathology appeared Mogensen DN Ⅰ,Ⅱ or / and Ⅲ respectively.The treatment lasted 12 week.Then CEUS was performed on bilateral kidney to observe the renal perfusion,the parameters included:curve ascending slope(K1),time to peak intensity(PIT),peak signal intensity(PSI),area under the curve(AUC) and time to half of peak intensity(HPT).Results At the end of 12 weeks,compared with control group,PIT and HPT delayed,PSI and K1 decreased in untreatment group and group C,AUC of untreatment group increased.Compared with untreatment group,PIT,HPT and AUC decreased,PSI and K1 increased in group A,B and C.Compared with group C,HPT of treatment group A,B decreased.Compared with control group,renal's size increased and the cortex was thickened in untreatment group and group A,B,C.The pathological examination showed:renal were damaged more seriously in untreatment group than that in group A and B.There was no significant difference in Scr and BUN between all groups.Conclusions Assessment of CEUS for renal perfusion of diabetic mellitus rabbits at early stage is feasible and captopril can improve renal perfusion of DM rabbits.

Objective To assess the renal perfusion of different severity of hepatic cirrhosis by contrast-enhanced ultrasound with time-intensity curve.Methods Forty hepatic cirrhosis patients with normal conventional renal function enrolled the study and were subdivided into Child-Pugh A group ( n =14),Child-Pugh B group ( n =14) and Child-Pugh C group ( n =12).Thirty healthy subjects were selected as the control group.The parameters of perfusion were compared among the groups.The correlations between parameters and score of Child-Pugh were analyzed using Pearson correlation.Results The cirrhosis groups showed a decrease of peak intensity,increase of time to peak,and reduce of area under the curve in the renal cortex( P ＜0.05).Compared with Child-Pugh A group,the peak intensity decreased in both ChildPugh B and C groups,time to peak increased in Child-Pugh C group( P ＜0.05 or P ＜0.01 ).The peak intensity had negative correlation with Child-Pugh score( r =- 0.506,P ＜0.01 ).Conclusions Contrastenhanced ultrasound combined with time-intensity curve can assess renal perfusion of hepatic cirrhosis quantitatively.The peak intensity was correlated to the severity of cirrhosis.

Objective To explore the clinical features of hepatolenticular degeneration (HLD) with hemolytic anemia as the first manifestation, and its relationship with ichthyosis.Method The clinical manifestations, course, diagnosis, and treatment of HLD combined with ichthyosis in one child who presented with hemolytic anemia as the ifrst manifestation, were retrospectively analyzed.Results In ten-year-old girl, the ifrst symptom was fatigue and jaundice, and then progressed to abdominal distension 5 days after admission. The reticulocyte percentage was 8.5%. Coombs test was negative. The K-F rings appeared on the cornea. The CT scan of Abdomen showed pleural and peritoneal effusion, abnormal substance deposition in liver and splenomegaly. There were mutations inATP7B andTGM1 genes, both of which were located on fourteenth of chromosomes. The diagnosis of HLD combined with ichthyosis was conifrmed. After symptomatic and supportive treatments such as removal of copper, prevention of the copper absorption and supplement of vitamin A, the disease was brought to remission.Conclusion HLD with hemolytic anemia as the ifrst manifestation, combined with ichthyosis was extremely rare.