BACKGROUND: In the process of limb allograft, apoptosis of target cell is one of the main mechanisms of dysfunction of allograft, which might lead to the failure of allotransplantation. It is assumed that immunosuppressant may relate with cell apoptosis. OBJECTIVE: To investigate effects of FK506 on Bcl-2 mRNA and Bax mRNA expressions and cell apoptosis in rat limb allograft. DESIGN, TIME AND SETTING: The randomized controlled animal trial was performed at the Animal Experimental Center of First Hospital of Harbin Medical University from June 2005 to November 2006. MATERIALS: Fifty-six clean-grade healthy male SD recipient rats and 56 Wistar donor rats were selected. FK506 was product of Fujisawa, Japan (No. 100143G). METHODS: Right hind limb was separated from the upper segment of thigh of SD rat (donor), and washed using heparin saline. The recipient rates underwent limb allotransplantation from allogenetic Wistar to establish injury model. The recipients were randomly divided into two groups (n=28): immunosuppressant group was injected with FK506 1 mg/kg per day, and the control group was not given any immunosuppressant. The right hind limb including skin, subcutaneous tissues, muscles and femoral arteriolar-venular tissue mass were harvested on postoperative days 1, 3, 5, and 7. MAIN OUTCOME MEASURES: Bcl-2 Mrna and Bax mRNA expression were detected using reverse transcription polymerase chain reaction (RT-PCR), and cell apoptosis was detected using in situ terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling technique. RESULTS: Fifty-six model rats were included in final analysis. On the 3, 5, and 7 days after FK506 injection, Bcl-2 mRNA expression in immunosuppressant group was significantly higher than the control group (t=7.18-21.20, P

Objective To investigate serum NK cell levels in HBeAg positive early pregnancy women with immune activation.Methods Fifty four HBeAg positive pregnant women admitted in Taizhou People's Hospital from September 2010 to April 2013 were enrolled in the study.Among them,the serum HBV DNA load decreased ≥2 log at 12 weeks after pregnancy in 24 cases (immune activation group) and HBV DNA did not decrease in 30 cases (immune tolerance group).The serum level of alanine aminotransferase (ALT),total bilirubin,HBeAg,HBV DNA load and NK cells were measured.Results At week 12 of gestation,the mean ALT levels and ALT abnormality rate in immune active group were higher than those in immune tolerance group [(146.7 ±93.1) vs.(44.1 ± 14.7) U/L,t =2.95,P＜0.05,50.0％ vs.6.7％,x2 =4.97,P ＜0.05].There was no significant difference of HBeAg level between two groups before pregnancy,while HBeAg level in immune activation group was lower than that in immune tolerance group at week 12 week of gestation [(291.8 ± 170.5) vs.(443.7 ± 289.9) S/CO,t =2.81,P ＜0.05].The percentage and absolute number of NK cells in immune activation group were higher than those in immune tolerance group [(26.7 ±9.1)％ vs.(17.1 ±7.8)％,t =2.52,P ＜0.05 and (370.9 ±136.4)/μl vs.(213.2 ±97.8)/μl,t =2.38,P ＜0.05,respectively].Conclusions In HBeAg positive early pregnant women with immune activation,the inhibition of HBV DNA might be associated with the activation of NK cells.

Objective To observe the clinical efficacy of glucocorticoidcombined with the eradication of helicobactepylori (HP) in the treatmenof idiopathithrombocytopenipurpura(ITP) and itinfluence on platelecounand cell subset.MethodNinety-foucaseof ITP were randomly divided into two group.The control group(47 cases) wagiven prednisone treatmen, while the observation group(47 cases) wagiven prednisone combined with HP eradication treatmen.The clinical effecand recur-rence rate within 1 yeawere observed ,and the changeof platelecounand cell subsetwere tested .ResultThe total effective rate of the observation group wa89 .36% ,which wasignificantly highethan 70 .21% in the control group ;the 1-yearecurrence rate in the observation group wa36 .17% ,which wasignificantly lowethan 72 .34% in the control group ,the differencebetween the two grouphad statistical significance (P＜0 .05) .The platelecounaftetreatmenin the two groupwere significantly in-creased ,platelet-associated antibody(PAIgG) level wasignificantly decreased ,buthe improvemenin the observation group wamore significant(P＜0 .05) .The percentage of CD3+CD4+ % increased significantly ,the percentage of CD3+CD8+ % wadecreased significantly ,buthe improvemenin the observation group wamore obviou(P＜0 .05) .Conclusion Glucocorticoidcombined with HP eradication treatmenin treating ITP hadefinitely clinical efficacy ,can increase the platelecoun,improve cell subsetimbalance ,and iworthy to be clinically promoted .

Background and purpose:The time from ifrst onset of symptoms or signs to a deifnitive diagnosis is deifned as diagnostic interval (DI). The relation of DI to other clinicopathological parameters andthe impact of DI on prognosis of patients with triple-negative breast cancer (TNBC) remain unclear.This article plans to make an intensive study of these questions.Methods:The clinical records of a series of 83 consecutively presenting unselected patients referred to the Shanghai Sixth People’s Hospital with diagnosed TNBC between September 2009 and September 2015 were retrospectively reviewed. Clinical and pathological factors included were investigated by univariate analysis using the Kaplan-Meier method, the factors associated with prognosis were further evaluated by multivariable analysis with Cox progression model.t-test and Kruskal-Wallis test were used to study the correlation between DI and other characters.Results:DI: stage T3>T1 (P=0.01), stageⅢ>Ⅱ (P=0.03) andⅠ (P=0.01). Compared with patients of DI≥3 months, the <3 months group had earlier age (P=0.028) and TNM stage (P=0.035). T stage, N stage, neoadjuvant chemotherapy, TNM stage and DI are inlfuencing factors of overall survival (OS). Age, T stage, N stage, TNM stage, menstrual status and neoadjuvant chemotherapy are inlfuencing factors of progression-free survival (PFS). TNM staging is an independent inlfuencing factor for OS and PFS.Conclusion:Patients with later disease stage were more likely to have a longer DI; The shorter DI, the earlier age and stage of disease; DI is the inlfuence factor of OS; TNM stage is an independent inlfuencing factor for OS and PFS.

Objective To investigate the difference of amplitude of low-frequency fluctuation (ALFF) and fraction of amplitude of low-frequency fluctuation(fALFF) between Alzheimer's disease (AD)patients and normal aging (NA) controls by voxel-based analysis.Methods Thirty-one AD patients and 44 NA controls were enrolled in the study.Blood oxygen level dependent functional (BOLD) EPI data were obtained during resting-state by using 32-channel head coil.Data were realigned,normalized and then smoothed with 8 mm FWHM kernel.Resting-state fMRI toolkit(version 1.6) was used to generate ALFF and fALFF images.Independent two sample t-test was performed with SPM5 to compare ALFF and fALFF of AD and NA controls.Pearson correlation analysis was performed to examine the relationship between MMSE score and ALFF,fALFF parameters.The significance level was set to be uncorrected O.001 on the voxel level and 0.05 on the cluster level.Results AD patients showed increased ALFF in left temporal lobe (0.492 ±0.119) and right cingulated cortex (0.434 ± 0.093) of AD patients,which were 0.443 ± 0.068 and 0.380 ±0.081 in NA controls (t =2.658,2.227,P ＜ 0.05).Decreased fALFF was found in bilateral posterior cingulate cortices (1.167 ± 0.203) and increased fALFF was found in bilateral temporal lobes (left 1.226 ±0.127,right 1.146 ±0.214) with left side dominance,which were 1.453 ±0.269,1.134 ±0.088,1.014 ± O.132 in NA controls (t =5.001,3.695,3.285,P ＜ 0.05).Bilateral temporal ALFF and fALFF correlated with MMSE positively (r =0.768—0.909,P ＜ 0.05) with left dominance.Conclusion AD patients showed increased resting-state functional MRI changes correlated with MMSE score in the temporal lobes with left dominance,which indicated left temporal lobe may be the best location for the observation of disease progression in AD patients.

Objective To explore the mechanism by which the anti-human P185erbB2 scFv-Fc-IL-2(HFI)modulates tumor surface molecules and activates immune effector cells in vitro. MethodsMTT assay was used to test the proliferation and the LAK-like cytotoxicity. Flow cytometry assay was used to test the expression of ICAM-1, Fas and erbB2 receptors in tumor cells and the expression levels of CD molecules FasL and LFA-1 in human PBMC. Antibody-dependent cell-mediated cytotoxicity(ADCC)mediated by HFI against SKOV3, MCF-7 and SGC-7901 tumor cells was explored hv LDH release assay. Results The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI were upregulated, from 24.85％ and 0.53％ to 85.36％ and 59.19％ respectively, while the expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI were downregulated, from 98.48％, 42.60％ and 36.66％ to 94.01％,30.95％ and 12.36％ respectively. HFI could significantly enhance the proliferation activity of human PBMC, and CD3+ CD8+ T cells and CD3- CD16+ CD56+ NK cells were elevated, from 24.37％ and 6.90％ to 38.80％ and 13.45％ respectively. The expression levels of CD25, LFA-1 and FasL were significantly enhanced from 3.99％, 86.52％ and 5.02％ to 12.96％, 99.06％ and 16.19％. The LAK-like cytotoxicity of human PBMC treated with HFI against SKOV3, MCF-7,SGC-7901 tumor cells was significantly improved:HFI was effective in mediating ADCC against SKOV3,MCF-7 and SGC-7901 tumor cells which expressed high,medium and low levels of erbB2,respectively,and HFI-induced ADCC was correlated with the degrees of erbB2 expression on the tumor cells. Conclusion The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI are significantly upregulated. The expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI are downregulated. HFI can significantly enhance the proliferation activity of human PBMC. The LAK-like eytotoxicity of human PBMC treated with HFI against tumor cells is significantly enhanced. HFI iS effective in mediating ADCC and the activity of HFI-induced ADCC is correlated with the degrees of erbB2 expression on the tumor cells.

ObjectiveTo evaluate the app li cation value of the Dipstick Dye Immuno assay (DDIA) for screening chemotherapy targets of schistosomiasis in a lower endemic area. Methods[ WT5”BZ]In a lower endemic area of schistosomiasis a random sample of 463 individuals from a natural village were examined using miracidium hatching metho d, Kato Katz's method, DDIA, DGS COPT and ELISA. The positive rates of these a ss ays were compared. ResultsThe positive rate of stool examination was 3.9% in 463 individuals. The positive rate of DDIA was 15 8%. The positive rate in 18 stool positive subjects was 94 4% with Youden In dex 0 81. The positive rate of DGS COPT was 8 9% . The positive rate in 18 stool po sitive subjects was 72 2% with Youden Index 0 66. The positive rate of ELISA w as 18 4%. The positive rate in 18 stool positive subjects was 83 3% with Youden In dex 0 68. ConclusionDDIA was more suitable for application in screening target population in lower endemic areas than other im munoassys.

[ ABSTRACT] AIM:To study the expression of WNT5B in the breast cancer and further to discuss the correlation between WNT5B and clinicopathologic characteristics of breast cancer.METHODS:The expression of WNT5B at mRNA and protein levels was measured by real-time PCR and Western blot in 67 cases of breast cancer and the tissue adjacent to carcinoma.In addition, the immunohistochemical method was used to detect the expression of WNT5B in the breast cancer and the tissue adjacent to carcinoma.The relationships between WNT5B expression and clinicopathologic indexes were also analyzed.RESULTS:The expression of WNT5B in the breast cancer was obviously lower than that in the tissue adjacent to carcinoma (P<0.05).The expression of WNT5B at mRNA and protein levels in 67 samples of breast cancer was in va-rious degrees.The expression of WNT5B in T≤20 mm group of human breast cancer was obviously higher than that in T>20 mm group (P<0.05).The expression of WNT5B had no obvious correlation with axillary lymph node metastasis, histo-logical grade and immunohistochemical indexes of ER, PR, c-ErBb-2, p53 and Ki67 ( P>0.05) in the breast cancer. CONCLUSION:The expression of WNT5B decreases obviously in breast cancer.The expression of WNT5B is related to primary tumor size, which provides new ideas for the diagnosis and treatment of breast cancer, suggesting that WNT5B may be a new molecular marker for prognosis of breast cancer.

AIM:To explore the effect of HbA1c level in the patients with type 2 diabetes mellitus (T2DM) on the rheological properties of erythrocytes and the structure of hemoglobin (Hb).METHODS:The patients with T2DM were classified into 3 groups:the patients with good glycaemic control (group A, HbA1c<7.0%), the patients with poor glycaemic control (group B, 7.0%≤HbA1c<9.0%) and the patients with persistent hyperglycemia (group C, HbA1c≥9.0%) .The rheological properties of a single living erythrocyte were analyzed by the techniques of static imaging and anal -ysis.Ultraviolet-visible spectroscopy was used to study the structure of Hb .RESULTS: Compared with group H , the roundness effect factor (REF) of erythrocytes, which was positively related with HbA1c level, was significantly increased in groups A, B and C (P<0.05).The contact area of erythrocytes in group B and all the morphological parameters of erythrocytes in group C were significantly higher than those in group H ( P <0.05 ) .There were significant differences among groups A , B and C in deformation capacity and elastic parameters of the cell membrane ( P<0.05 ) , but with no difference in the long axis and short axis between group A and group B .Compared with group H , no obvious change in the spectral pattern and spectrum peak of Hb in groups A , B and C was observed.However, the absorbance of Hb, which showed a trend of gradual decline with the increase in HbA 1c level, in group B and group C was significantly decreased as compared with group H (P<0.01), and the Hb absorbance in group C were also lower than that in group A (P<0.05). CONCLUSION:With the increase in HbA1c level, the morphology along with the deformation function of erythrocytes in T2DM changes and declines gradually , and the structure of Hb may also change .

The hapten of Flumequine(FLU) with four carbon atoms spacer arm(FLUABA) was synthesized and coupled to bovine serum albumin(BSA) as immunogen using activated ester method. Balb/c mice were immunized by the artificial immunogen and the splenocytes of immunized mice were fused with Sp2/0 cells to obtain the monoclonal antibody(McAbs). A hybridoma cell line(DB6-E7) secreting anti-flumequine McAbs was obtained by limited dilution method and screened by indirect enzyme-linked immunosorbent assay(ELISA) using heterogenous coating antigen. The results showed that the subtype of the McAb was IgG_1, and the affinity was 8.19×10~8 L/mol. The haptens of FLU, FLUABA and FLUACA, with different space arm, were separately linked to ovalbumin(OVA) for heterologous or homologous coating antigen. The results of indirect ELISA and indirect competitive ELISA(icELISA) indicated that the heterologous coating antigen could improve the sensitivity of ELISA significantly. By heterologous coating antigen(FLU-OVA), the icELISA showed an IC_(50) value of 26.33 μg/L, LOD of 4.0 μg/L, and the workable range of 8-114 μg/L (IC_(20)-IC_(80)). Cross-reactivity studies showed that the McAbs were quiet specific for FLU, no cross-reactivity(<0.1%) was detected between the obtained McAbs and the quinolones compounds or other structural similarity compounds. The developed icELISA for FLU can satisfy the detection criteria of flumequine in animal food-products.