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.

12E7 Antigen ; Aged ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Diagnosis, Differential ; Giant Cell Tumors ; pathology ; Giant Cells ; pathology ; Humans ; Male ; Mesenchymoma ; pathology ; Mucin-1 ; metabolism ; Myositis Ossificans ; pathology ; Osteosarcoma ; metabolism ; pathology ; surgery ; Penile Neoplasms ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Vimentin ; metabolism

ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Calnexin ; metabolism ; Calreticulin ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; virology ; Cysteine Endopeptidases ; metabolism ; Female ; HLA-A Antigens ; metabolism ; Human papillomavirus 16 ; isolation & purification ; Humans ; Lymphatic Metastasis ; Papillomavirus Infections ; Proteasome Endopeptidase Complex ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology ; virology

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

Objective:To investigate the mechanism of liver and lung injury in mouse septic models.Methods:Twenty-four male Kunming mice were subjected to cecal ligation and puncture(CLP)or sham operation.The permeability of microvasculature,water contents,activities of myeloperoxidase(MPO)and the apoptosis of microvascular endothelial cells in lung microvasculature and liver sinus were examined 3 h and 12 h after operation.Results:Both the liver and lung showed a significant increase in microvessel permeability at 12 h in CLP group compared with sham operation group.MPO activity and water content in CLP group were obviously higher than those in the sham operation group.The apoptosis of lung microvascular endothelial cells at 12 h in CLP group(5.03?0.92)% was significantly higher than that of control group(3.48?1.21)%(P

To optimize the water extraction process for glycyrrhiza. Methods: HPLC was used to determine the con-tents of glycyrrhizic acid and liquiritin. The comprehensive index included the contents of glycyrrhizic acid and liquiritin and the yield of dry extract. The water amount and the extraction time were selected as the independent variables, and the comprehensive index was set as the dependent variable. Design-expert 8. 06 software was used to fit multivariate linear or quadratic multinomial models for the experimental values. Response surface methodology was used to optimize the water extraction process. The prediction was carried out through comparing the observed and predicted values. Results:The regression coefficient of binomial fitting complex model was as high as 0. 979 7. The optimum conditions of extraction process were as follows:12-fold amount of water, extracting 3 times with 90 min for each time. The deviation between the observed and predicted values was -1. 72%. Conclusion: Central composite design-response surface methodology is convenient and highly predictive in optimizing the water extraction process for glycyrrhiza, which can be applied in the further membrane separation and purification.

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

OBJECTIVETo explore the potential role of miR-195 on invasiveness and prognosis of breast cancer.

METHODSThe RNA in formalin-fixed paraffin embedded (FFPE) of 88 breast cancer patients with primary tumors was extracted, and miR-195 levels were quantified by quantitative real-time polymerase chain reaction (real-time PCR). The relationship of miR-195 levels and clinicopathological variables were assessed by Mann Whitney-U test. Recurrence-free survival and overall survival curves were derived from Kaplan-Meier estimates and the curves were compared by Log-rank tests. Cox regression analysis was used for multivariate analysis. All statistical tests were two-sided.

RESULTSThe levels of miR-195 in the breast cancer with histological high grade, tumor size of T3-4, lymph nodal involvement or vessel invasion were significantly down-regulated, compared with those of patients with histological low grade (Z = -2.271, P = 0.023), tumor size of T1-2 (Z = -2.687, P = 0.007), no lymph node metastasis (Z = -1.967, P = 0.049) and vessel invasion (Z = -2.432, P = 0.015). In addition, no statistically significant difference (P > 0.05) was identified between miR-195 levels and hormone receptors status, HER-2 expression, TNM stage, tumor types, recurrence and menstrual status. When considering 2(-ΔCt) = 0.270 (median level) as cut-off value, Kaplan-Meier survival analysis indicated that patients with high miR-195 level showed a positive association towards a longer survival, either recurrence-free survival (χ(2) = 5.985, P = 0.014) or overall survival (χ(2) = 30.05, P = 0.000). In a multivariate analysis, miR-195 expression on FFPE correlated significantly with outcomes of breast cancer (HR = 0.040, 95%CI: 0.009 - 0.179, P = 0.000) and was independent of other prognostic factors.

CONCLUSIONSIt suggests that miR-195 expression on FFPE is inversely correlated with histological high grade, bigger tumor size, lymph node involvement, vessel invasion. Furthermore, as independent prognostic factor, low miR-195 significantly contributes to poor outcomes of breast cancer.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; pathology ; Female ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; MicroRNAs ; genetics ; Middle Aged ; Multivariate Analysis ; Prognosis ; RNA, Neoplasm ; genetics

Objective To evaluate the applied value of percutaneous oxygen-ozone injection in the treatment of lumbar disc herniation under open 0.23 T MRI guidance.Methods Mounted with ipath 200 optical tracking system,MR-guided injection of oxygen-ozone were performed via a medial border of the articular processes approach in 73 patients with clinically diagnosed LDH.MR compatible 19.5G or 21.0 G biopsy needle was used. Discography was performed in order to select indication before injection oxygen- ozone into nucleus pulposus in 26 patients.Sixty-four patients were injected to three sites:(1)Six to 10 ml oxygen-ozone was injected into discs centers,injected and suctioned alternately in order to make nucleus pulposus oxidation thoroughly.(2)The needle was withdrawn according to the scale of biopsy needle and optical tracking.Then,10 ml oxygen-ozone was injected into disc herniation. (3)After that,needle was withdrawn further about 1.0—1.5 cm to outside of annulus fibrosus.Fifteen to 20 ml oxygen-ozone was injected into intervertebral foramina around nerve roots.The oxygen-ozone concentration was 35—45?g/ml. Nine patients were only performed injection of oxygen-ozone into around nerve root,while not injection oxygen-ozone to nucleus pulposus for considering bad curative effect after discography.Results All of 73 patients were successfully local targeted and treated under MRI guidance without serious complications, such as nerve root injury.After 3—6 months follow-up,total overall efficacy was 91.3% with the excellent in 28,good in 39,and poor in 6,respectively.Conclusion Open MR-guided injection of oxygen-ozone, mounted with optical tracking system,is a safe and effective minimally invasive therapy for treating LDH.