In this study ,we intended to prepare anti‐Toxoplasma gondii aquaporin (TgAQP) peptide antibody which was used to the application in the detection of the aquaporin expression and its subcellular localization of Toxop lasma gondii (T .gondii) ME49 strain .The B cell peptide antigen was designed based on the TgAQP amino acids sequence .After the pep‐tide antigen was conjugated to the KLH ,the fusion antigen was injected into New Zealand rabbits to prepare polyclonal anti‐body ,followed by identification of ELISA ,Western‐blotting and immunofluorescence assays .The ELISA showed that the titer of anti‐TgAQP antibody was about 1∶40 000 .Western blotting revealed the specific affinity of the antigen to polyclonal anti‐body at 29 .9 kDa protein T .gondii .The protein detected by the indirect immunofluorescence assays was distributed in the cy‐toplasm of the parasite .Thus far ,the anti‐TgAQP polyclonal antibody was successfully prepared ,providing a useful tool for further study of biological function and metabolic characteristics of TgAQP .

Epithelial-to-mesenchymal transition (EMT) plays a critical role in cancer metastasis,and is relevant to the inflammatory microenvironment.Lipopolysaccharide (LPS),a cell wall constituent of gram-negative bacteria,has been reported to induce EMT of cancer cells through TLR4 signal.We previously reported that LPS promoted metastasis of mesenchymallike breast cancer cells with high expression of cyclin D 1 b.However,the role of cyclin D1b in LPS-induced EMT has not been fully elucidated.In the present study,we described that cyclin D1b augmented EMT induced by LPS in MCF-7 breast cancer cells.Cyclin D1b markedly amplified integrin αvβ3 expression,which was further up-regulated under LPS stimulation.Our results showed ectopic expression of cyclin D1b promoted invasiveness of epithelial-like MCF-7 cells under LPS stimulation.Additionally,LPS-induced metastasis and EMT in MCF-7-D1b cells might depend on αvβ3 expression.Further exploration indicated that cyclin D1b cooperated with HoxD3,a transcription factor promoting αvβ3 expression,to promote LPS-induced EMT.Knockout of HoxD3 repressed LPS-induced EMT and αvβ3 over-expression in MCF-7 cells with high expression of cyclin D1b.Specifically,all these effects were in a cyclin D1a independent manner.Taken all together,LPS up-regulated integrin αvβ3 expression in MCF-7 cells with high expression of cyclin D 1b and induced EMT in breast cancer cells,which highlights that cyclin D1b may act as an endogenous pathway participating in exogenous signal inducing EMT in breast cancer cells.

Objective To evaluate the significance of human interleukin-31(IL-31)in the pathogenesis of atopic dermatitis and its correlation with pruritus in patients with atopic dermatitis(AD).Methods Twenty-two children with mild to severe atopic dermatitis and 22 age-matched healthy controls were included in this study.Patients and controls were randomly and equally assigned into stimulation and non-stimulation groups.Venous blood samples were obtained from all participants,peripheral blood mononuclear cells were isolated from these samples and cultured with(stimulation groups)or without(non-stimulation groups)staphylococcal enterotoxin B(SEB)for 24 hours.Then,the mRNA expression of IL-31 on PBMCs was assessed via real-time reverse transcription-PCR.ELISA was used to detect the total serum IgE level in these objects.The severity of AD in patients was rated according to scoring atopic dermatitis(SCORAD).The relationship between the mRNA expression of IL-31 and the level of serum total IgE.severity of atopic dermatitis,and degree of pruritus.was evaluated.Results The expression of IL-31 mRNA on non-stimulated PBMCs from patients was 23.2 folds as high as that from the healthy controls(P＜0.01).The stimulation with SEB upregulated the mRNA expression of IL-31 on PBMCs.and the increase on PBMCs from patients was 20.44 times of that from the controls.The total serum IgE level was 260.05 IU/mL(5.9-1131.01 IU/mL)and 17.7 IU/mL(5-140.7 IU/mL)in the Patients and controls respectively(P＜0.01).There was no significant correlation between the mRNA expression of IL-31 and disease severity or total serum IgE level(r=0.07.0.22respectively.both P＞0.05)in patients witll AD.Condusions IL.3 1 is involved in t11e pathogenesis of AD,which is unlikely to be IgE-dependent.SEB can induce the rapid expression of IL-31 on PBMCs of healthy human,and is an important modulator for the production of IL-31.

Objective To investigate the value of ultrasonic elastography(UE) in diagnosing breast small masses (diameter≤10 mm). Methods The static sonograms of conventional ultrasound(US) and UE of the 274 breast small lesions which pathologically proved were reviewed. Images were assigned according to the B1-RADS criteria for US. The elastographic images were evaluated using improved 5-scoring system.Pathology was followed up as diagnostic criteria. The diagnostic performance of the 2 methods was evaluated with receiver operating characteristic curve(ROC). Results The areas under the ROC curve of the two modalities were 0. 891 (UE) and 0.742 (BI-RADS), respectively. The difference between them was significant. Conclusions UE is helpful in differentiating between malignant and benign breast small lesions.

ObjectiveTo evaluate the therapeutic efficacy and prognostic factors of intensity-modulated radiotherapy combined with chemotherapy for early-stage nasopharyngeal carcinoma patients in northwest China. MethodsFrom January 2006 to December 2009,58 patients with early-stage nasopharyngeal carcinoma were treated with IMRT in Xijing hospital,the clinical data were analyzed retrospectively.Survival rates was calculated by the Kaplan-Meier method and the differences was compared by the Logrank test.Univariate analysis method was use to identify all significant factors.ResultsThe follow-up rate was 100％.The follow-up time of 46 patients was more than 3 years.The 1-,2 and 3-year survival were 98％,94％ and 91％,respectively.The 3-year overall survival (OS),local recurrence-free survival (LRFS),distant metastasis-free surv ival (DMFS) for T1N0-1,T2N0 and T2N1 stage were 100％,100％,100％ and 74 ％,81％,87 ％,respectively ( x2 =5.74,P =0.01 ; x2 =4.95,P =0.03 ; x2 =4.24,P=0.04).The 3-year OS,LRFS,DMFS for IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 85％,85％,88％ respectively ( x2 =4.02,P =0.04; x2 =4.12,P =0.03 ; x2 =4.84,P =0.02).In T2N1 stage,IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 79％,79％,80％ respectively (x2 =5.28,P =0.03 ;x2 =4.84,P =0.04;x2 =4.72,P =0.04).In univariate analysis,N stage,clinical stage,IMRT combined with chemotherapy were significantly associated with the survival ( x2 =5.39,P =0.02 ; x2 =5.74,P =0.01 ; x2 =4.02,P =0.04).Conclusions In all early-stage nasopharyngeal carcinoma,T2N1 stage is a sub-group of high risk of distant metastasis.Combination of IMRT and chemotherapy may improve the LRFS,DMFS and OS in those patients.

Objective To summarize the clinical features,diagnosis and treatment of autologous urologic neoplasms in renal transplant recipients.Methods A retrospective analysis on the clinical data of 25 renal transplant recipients was done in our center.The onset time of new neoplasms was between 29 to 72 months after transplantation,with an average of 48.2 months.Intermittent hematuria was the first symptom in 23 patients,and the rest two cases were diagnosed through routine examination. The pathological diagnoses of thee cases were renal carcinoma,which were treated by transperitoneal laparoscopic radical nephrectomy.Eight cases were diagnosed as having renal pelvic tumor,which was treated by radical resection for the renal pelvic carcinoma.Fourteen cases were diagnosed as having bladder cancer,which was treated by transurethral resection of bladder tumor (13 cases) or radical cystectomy (one case).All patients were subjected to surgical treatment.The dosage of MMF,CSA/Tacrolimus was decreased to 1/2-2/3 of their original dosage. Sirolimus was used in place of calcineurin inhibitors in four patients.Immunosuppressive regimes and adjuvant therapy were given after surgery treatment.Results Twenty-five patients were followed up for 12-84 months.Contralateral renal carcinoma combined with lung and chest multiple metastases occurred in one case after radical nephrectomy,who died after targeted therapy 6 months later.Two patients with lymph node metastasis died 14 months and 20 months after surgery respectively.The rest 22 patients were closely followed up,whose creatinine remained 98-163μmol/L.Conclusion More attention should be paid to patients with hematuria after renal transplantation to screen the autologous urinary neoplasms.Patients should be treated with surgical procedures,and immunosuppressive regimens should be adjusted postoperatively.

Objective To study the correlation between drug-resistance of paclitaxel and the expres- sion levels of anti-apoptotic protein survivin and the role of matrine reversal resistance of PTX in non-small cell lung cancer.Methods The expressions of survivin in 120 samples of human lung cancer with paclitaxel chemotherapy were detected by immunohistochemical staining.Results The positive expression rate of sur- vivin was 57.5%.In the positive expression of survivin group,chemotherapy combined matrine could improve response rate and survival time(P0.05).Conclusion The survivin expression was related to the response rate of paclitaxel in lung cancer.It might be a valuable new marker to predict the drug-resistance of paclitaxel.In this prospective research,survivin protein,which promoted the apoptosis caused by paclitaxel,may be the target of martine.It could partly reverse the drug re- sistance to paclitaxel.

Background and purpose:LMP1 was one of the protein encoded by EBV latent gene,which was found to be able to transform cell lines and alter the phenotype of cells due to its oncogenic potential.In human epithelial cells,LMP1 alters many functional properties that are involved in tumor progression and invasions.In this study we investigated the influence of LMP1 silence on AP-1 signal transduction pathway and its downstream factors involved with cell transformation,proliferation and apoptosis.Methods:The chemically synthetic siRNA targeting LMP1 was transfected into EBV positive gastric carcinoma epithelial cell line by lipofectamine 2000 at 50 nmol/L final concentration.The protein expression of c-Jun,JunB and CDK4 was tested by,Western blotting.The mRNA of c-Jun,surviving,CDK4 and MMP9 mRNA were tested by RT-PCR.The expression of survivin were tested by immunohistochemistry.Results:Compared with the cell control,CDK4 mRNA was up regulated(P

AIM: To investigate the expression of voltage-gated chloride channels (ClC)-3 protein and mRNA in human glioma specimen and its biological function. METHODS: The expression of C1C-3 was observed by immunohistochemical staining in 24 cases of human glioma, 4 cases of brain metastic cancer specimens and 5 cases of normal brain tissue as control; The C1C-3 mRNA expression were detected in the specimens with positive expression of ClC-3 protein by RT-PCR. RESULTS: ClC-3 protein was found negative in 4 cases of normal brain tissues and positive in 19 cases of human glioma and 4 cases of brain metastic cancer specimens. ClC-3 protein was mainly expressed in the membrane or cytoplasm of neoplastic cells and microvascular endothelial cells. The expression of ClC-3 mRNA was detected in 16 cases of human glioma and 4 cases of brain metastasis cancer specimens among the tissues with the positive expression of ClC-3 protein. The level of protein and RNA of ClC-3 in high malignant oligodendrogliomas was higher than that in low malignant ones. CONCLUSION: ClC-3 is generally expressed in human glioma and brain metastic cancer and is probably correlated with the classification of its pathological malignance.

Objective To summarize the clincical experience of transumbilical Laparoendoscopic Single-site (LESS) nephrectomy and to evaluate its safety and efficacy. Methods From December 2008 to August 2010, we have performed 20 cases of transumbilical LESS nephrectomy by Tri-Port system, of which 9 patients underwent LESS radical nephrectomy (left 8, right 1, stage T1 ), 1 patient underwent LESS radical resection of right ureteral carcinoma, 10 patients underwent LESS simple nephrectomy (left 5, right 5). The Tri-Port system was inserted transperitoneally through a 2 cm umbilical incision. A 5-mm 30° telescope was introduced through the port to visualize the operative field. Flexible equipment and standard laparoscopic equipment were used to perform the procedures.The incisions were extended to about 6cm in order to remove the specimens. Results Conversion to open surgery was necessary in one LESS radical resection of right ureteral carcinoma and one LESS simple nephrectomy, while the remaining 18 cases were successful (the addition of a single 5-mm port was necessary in 2 cases of LESS radical nephrectomy). The mean operative time was 197 min (85-510 min), mean estimated blood loss was 126 ml (50-400 ml), without blood transfusion in the perioperative period, mean postoperative hospital stay was 6.3 d (3-14 d), and mean duration of catheter drainage was 3.6 d (0- 14 d). Conclusions Transumbilical LESS nephrectomy is feasible, safe,minimally invasive and cosmetic. Long-term follow-up and a clinical control study are needed for evaluating clinical outcomes.