Objective To explore the effect of Mycobacterium tuberculosis antigen (Mtb-Ag) on neutrophils apoptosis.Methods The fresh isolated neutrophils from healthy adults blood were cultured with Mtb-Ag for 24 h,with or without pretreatment of nuclear factor -?B (NF-?B) inhibitor N-tosyl-L-phenylanyl chloromethyl ketone (TPCK) for 30 minutes.Annexin V staining and Flow cytometry were used to measure cell apoptosis of neutrophils.NF-?B DNA binding was measured by gelelectrophorestic mobility shift assay (EMSA) in neutrophils after incubated with Mtb-Ag for 0,1,2,4,6,24 hours.Results Comparing to the spontaneous apoptosis (55%?6%) of neutrophils after culture in vitro for 24 h,treatment of Mtb-Ag (1.125 mg/ml) decreased the cell apoptosis of neutrophils (32%?3%).The NF-?B shift bands were detected at 1 h in neutrophils after stimulated by Mtb-Ag,and reached maximum peak at 2 hours,and then returned to basal levels within 24 h.Pretreatment of TPCK inhibited the anti-apoptosis role of Mtb-Ag in neutrophils.Conclusion Mtb-Ag prevents neutrophils apoptosis and its inhibitory role concerns NF-?B pathway.

Accidents, Occupational ; statistics & numerical data ; Acute Disease ; China ; epidemiology ; Humans ; Occupational Diseases ; epidemiology ; Poisoning ; epidemiology

Objective To explore the effect of Ferroprotin 1 expression on tumorigenesis,invasiveness and survival of breast cancer.Methods In this study,100 breast cancer patients were enrolled.IHC SP was used to detect the expression of Ferroprotinl in paraffin-embedded tissues.The `association of Ferroprotin 1 expression and clinico-pathological parameters was evaluated by chi-square test.Survival analysis was calculated by Kaplan-Meier model and Log-rank test.Results The expression of Ferroprotin 1 was significantly higher in para-cancerous normal tissues (37/100,37％ ) than that in breast cancer tissues (24/100,24％ ;P =0.046 ).In these with positive axillary LN,there were more with low expression level of Ferroprotin 1 ( 36/40,90％ ) than those with high expression level ( 4/40,10％ ),P =0.007.More patients with low Ferroprotin1 were at advanced stage than those with high ferroprotin1 [Ⅲ 44/57 (77.2％) ;Ⅳ 17/18 ( 94.4％ )]( P =0.05 ).No significant association was found between ferroprotin1 and tumor grade,histology type,ER/PR,HER2,tumor size (P＞0.05).Ferroprotin1 has no significant effect on breast cancer survival ( P =0.591 ) by Kaplan-Meier curve and Log-rank test.Conclusions Low Ferroprotin 1 may lead to the tumorigenesis of breast cancer.Downregulated Ferroprotin1 promotes the LN involvement of breast cancer and accompanies with more advanced disease.However Ferroprotinl might not play an important role in the survival of breast cancer.

According to the results of quantum chemistry calculation and the present research status in the relationship between the structures and the functions of DT, the E154 in DT catalyzing domain was mutated to aspartic acid and arginine in order to study the effects of the alteration on the biological activities. By means of gene site-direct mutation, two mutated genes were prepared and the high performance expression was obtained in E.coli system. The results of toxcity studies indicated that the acute toxicity in guinea pig and cytotoxicities of mutant E154D increased slightly in compared with those of recombination wild toxin, and contrarily, those of E154R decreased obviously.

Objective:To prepare highly purified Staphylococcal enterotoxin B(SEB) by affinity chromatography and test its activities.Methods:Anti-SEB McAb(1D2) purified by precipitation method with caprylic acid was coupled to Sepharose 4B. And then the SEB was isolated using an affinity chromatography column. In addition, we analyzed the superantigen activity and antigen activity of SEB.Results:The purification efficiency of SEB was 60.71% by affinity chromatography. Its purity was higher than those of standard preparation and the SEB purified by ion change chromatography. At the same time, the purified SEB by affinity chromatography possesses favourable activities of superantigen and antigen.Conclusion:McAb affinity chromatography could be used for purification of SEB with high efficiency.

Objective:To investigate the predictive value of tumor regression rate after induction chemotherapy for survival of patients with locally advanced nasopharyngeal carcinoma.Methods:A total of 161 patients with stage Ⅲ-ⅣA nasopharyngeal carcinoma newly diagnosed at the Daping Hospital of Army Medical University from January 2009 to December 2012 were selected as the research subjects. The relationships between tumor size changes before and after induction chemotherapy and survival time were analyzed. Kaplan-Meier method was used to draw the survival curve accompanied with log-rank test. Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with nasopharyngeal carcinoma.Results:There were statistically significant differences in the tumor regression rate of primary lesions between N 1and N 2-3( Z=2.177, P=0.029), T 1-2and T 3-4( Z=-4.501, P<0.001)patients after induction chemotherapy. In N 1stage patients, the 5-year overall survival (OS) rates of patients with primary lesions achieving objective response ( n=18) and those without objective response ( n=19) after induction chemotherapy were 88.89% and 57.45%, and patients with cervical lymph node metastatic lesions achieving objective response ( n=19) and those without objective response ( n=18) were 86.72% and 49.10% respectively, with statistically significant differences ( χ2=6.023, P=0.014; χ2=7.441, P=0.006). In N 2-3stage patients, the 5-year OS rates of patients with primary lesions achieving objective response ( n=81) and those without objective response ( n=43) after induction chemotherapy were 77.56% and 50.70%, and patients with cervical lymph node metastatic lesions achieving objective response ( n=85) and those without objective response ( n=39) were 75.11% and 52.04% respectively, with significant differences ( χ2=8.037, P=0.005; χ2=7.268, P=0.007). Univariate Cox regression analysis showed that in patients with stage N 1, the tumor regression rate of primary lesions ( HR=0.048, 95% CI: 0.004-0.644, P=0.022), the efficacy of primary lesions ( HR=0.174, 95% CI: 0.037-0.830, P=0.028), the efficacy of cervical lymph node metastatic lesions ( HR=0.154, 95% CI: 0.033-0.725, P=0.017) after induction chemotherapy were significantly associated with OS; in N 2-3stage patients, the tumor regression rate of primary lesions ( HR=0.178, 95% CI: 0.056-0.564, P=0.003), the tumor regression rate of cervical lymph node metastatic lesions ( HR=0.081, 95% CI: 0.020-0.324, P<0.001), the efficacy of primary lesions ( HR=0.422, 95% CI: 0.228-0.781, P=0.006), the efficacy of cervical lymph node metastatic lesions ( HR=0.439, 95% CI: 0.238-0.813, P=0.009) after induction chemotherapy were significantly associated with OS. In multivariate Cox regression including N stage and tumor regression rate, N stage and efficacy, the interaction items were not statistically significant (all P>0.05). In T 1-2stage patients, the 5-year OS rates of patients with primary lesions achieving objective response ( n=45) and those without objective response ( n=13) after induction chemotherapy were 77.55% and 84.62%, and patients with cervical lymph node metastatic lesions achieving objective response ( n=43) and those without objective response ( n=15) were 78.89% and 80.00% respectively, with no significant differences ( χ2=0.239, P=0.625; χ2=0.005, P=0.943); in T 3-4stage patients, the 5-year OS rates of patients with primary lesions achieving objective response ( n=54) and those without objective response ( n=49) after induction chemotherapy were 78.90% and 45.00%, and patients with cervical lymph node metastatic lesions achieving objective response ( n=61) and those without objective response ( n=42) were 75.10% and 42.89% respectively, with significant differences ( χ2=13.615, P<0.001; χ2=12.752, P<0.001). Univariate Cox regression analysis showed that in patients with stage T 1-2, the tumor regression rate, the efficacy of primary lesions and cervical lymph node metastatic lesions after induction chemotherapy were not related to OS (all P>0.05); in T 3-4stage patients, the tumor regression rate of primary lesions ( HR=0.121, 95% CI: 0.033-0.444, P=0.001), the tumor regression rate of cervical lymph node metastatic lesions ( HR=0.126, 95% CI: 0.036-0.442, P=0.001), the efficacy of primary lesions ( HR=0.297, 95% CI: 0.150-0.588, P<0.001), the efficacy of cervical lymph node metastatic lesions ( HR=0.329, 95% CI: 0.173-0.625, P=0.001) after induction chemotherapy were significantly associated with OS. Multivariate Cox regression analysis showed that the interaction test of T stage and the efficacy of primary lesion trended to be statistically significant ( P=0.062). Conclusion:In patients with stage Ⅲ-ⅣA nasopharyngeal carcinoma, the responsiveness to induction chemotherapy in stage T 3-4patients has important value in predicting survival prognosis.

BACKGROUND:Conventional serological, imaging and laboratory diagnosis methods for diagnosing periprosthetic infection are difficult to achieve ideal sensitivity, specificity and accuracy. In particular, the differential diagnosis of septic loosening and aseptic loosening in late stage after prosthetic replacement lacks of commonly accepted diagnostic criteria. The combination of preoperative, intraoperative and postoperative diagnostic methods is of great significance in the identifying infection diagnosis and formulating renovation strategy.
OBJECTIVE:To diagnose periprosthetic hip joint infection by combined use of preoperative 99Tcm triplephase bone scanning and intraoperative neutrophil count in frozen section analysis, and to compare with conventional diagnostic tools.
METHODS:Between April 2008 and May 2013, 44 hip revision cases were enrol ed. Combined with clinical symptoms and serological examination, 18 cases whose erythrocyte sedimentation rate and C-reactive protein increased were considered as clinical y suspected infection group;26 cases with normal serological examination were considered as clinical y suspected aseptic loosening group. 99Tcm triplephase bone scanning was performed in al patients, and combined with intraoperative frozen section analysis. If both results were negative, the diagnosis was aseptic loosening and did one-stage revision surgery;otherwise, periprosthetic infection was detected and two-stage revision surgery was conducted. By postoperative fol ow-up serological indexes and surgery outcomes, statistics of work curve of subjects was used to evaluate the diagnostic values of 99Tcm triplephase bone scanning and frozen section analysis.
RESULTS AND CONCLUSION:In 18 cases of clinical y suspected infection group, 16 cases of preoperative 99Tcm triplephase bone scanning and intraoperative frozen section results were positive, so two-stage revision surgeries were performed;2 cases with negative results received one-stage revision surgery after excluding infection. In 26 cases of clinical y suspected aseptic loosening group, 25 cases of preoperative 99Tcm three-phase bone scanning and intraoperative frozen section results were negative, and diagnosed as aseptic loosening, so one-stage revision surgery was performed. One patient affected infection of staphylococcus aureus in one-sample of the three samples in postoperative microbiological pathogen detection, and diagnosed as periprosthetic infection, so one-stage revision surgery was carried out. After combined use of 99Tcm triplephase bone scanning and intraoperative frozen section analysis, performance curve area increased from 0.906 to 0.972, sensitivity of diagnosis of infection increased from 89%to 94%, specificity of diagnosis of infection increased from 92%to 100%. However, no significant difference was detected in diagnostic methods. These data indicated that adding the method of 99Tcm triplephase bone scanning and intraoperative frozen section analysis technique, on the basis of serological and other conventional diagnostic tools, can differentiate infections from aseptic loosening of hip prosthesis. Negative results of 99Tcm triplephase bone scan showed high correlation between the diagnosis of aseptic loosening of the prosthesis and good diagnostic value to exclude infection. Combining with intraoperative frozen section method has a very good clinical value for the revision surgery.

Objective To summary and explore the indications,safety and effectiveness of no touch ostial cardioplegia delivery method during aortic valve replacement(AVR) for severe aortic insufficiency(AI),in order to provide guidance for clinical application.Methods Between September 2012 and February 2015,63 cases were randomly divided into two groups,in study group we used this technique in 31 patients with a mean age of(58.0 ± 15.0) years and a range of 31 to 74 years old.The degree of AI averaged (3.5 ± 0.5) according to a semiquantitative 5-grade scale[1].32 patients were enrolled in control group with a mean age of(60.4 ± 9.4) years and a range of 22 to 73 years old.The degree of AI averaged (3.6 ± 0.5).The abnormal aortic cusps were sutured though the aortic incision,which would be closed temporarily.Blood cardioplegia was infused repeatedly into the coronary osita through the closed aortic incision during the remainder of the procedure except AVR,when the closed aortic incision was reopened.Results In the study group,spontaneous resumption of nodal and normal sinus rhythm shortly after the removal of aortic cross-clamp was achieved in all patients,no hearts had to be electrically defibrillated and electrocardiogram showed no obvious S-T segment elevation.The time from cross-clamp removal to sinus rhythm was(12.3 ± 7.7) minutes in the study group,versus (18.6 ± 8.2) minutes in the control group(P =0.037).No operative mortality occurred in both groups.There was a significant increase in left ventricular ejection fraction[from(0.54 ± 0.09) preoperatively to(0.62 ± 0.06) postoperatively] (P =0.005) in patients of the study group,which was not seen in patients of the control group.Conclusion Preliminary results have shown that the no touch ostial cardioplegia delivery method is a feasible method for cardioplegia infusion during aortic valve replacement in patients with severe AI.

Objective To investigate the potential role of miR-34a on breast cancer recurrence and prognosis.Methods In this study,88 breast cancer patients underwent mastectomy with detailed clinical follow-up information.Extracting RNA from the formalin-fixed paraffin embedded samples,miR-34a levels were quantified by quantitative real-time polymerase chain reaction (qRT-PCR).miR-34a levels among clinico-pathological variables were accessed by Mann Whitney-U test.RFS and OS survival curves were derived from Kaplan-Meier estimates and the curves were compared by Log-rank tests.All statistical tests were two-sided.Results Significantly lower miR-34a level was found in tumor tissue compared to paired normal tissue (P =0.000).A potential relationship between miR-34a levels and existing clinico-pathological parameters of breast cancer,such as menstrual status,tumor size,nodal involvement,stage of disease,hormone receptor status,HER2 status,or tumor subtype was investigated.No statistically significant difference were identified for these features (P ＞ 0.05).miR-34a level was significant lower among G3 group than G1 + 2 group (P =0.024).Down-regulated miR-34a level was observed in breast cancer with later relapse compared to patients without relapse (P =0.008).When considering 2-△Ct =0.117 (median level)as cut-off value,patients with miR-34a up-regulation showed a positive association towards a longer survival,either RFS(P=0.026,Log-rank test) or OS(P =0.019,Log-rank test).Conclusions miR-34a,as a tumor suppressor,promotes differentiation and contributes to relapse when down-regulated.miR-34a has the potential as prognostic factor for breast cancer.

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation