Validation of the Prognostic Significance of the Prognostic Stage Group According to the Eighth Edition of American Cancer Joint Committee on Cancer Staging System in Triple-Negative Breast Cancer: An Analysis From Surveillance, Epidemiology, and End Results 18 Database.

BACKGROUND: The eighth edition of the American Cancer Joint Committee on Cancer (AJCC) staging system for breast cancer put forward the prognostic stage groups for the first time based on the traditional anatomic tumor-node-metastasis staging system. Our study intends to validate the predictive significance of the eighth edition staging system in triple-negative breast cancer (TNBC) patients. MATERIALS AND METHODS: We collected and accessed 26,589 eligible cases of TNBC from the Surveillance, Epidemiology, and End Results database (2010-2015) and reclassified the patient cohort according to the eighth edition of the AJCC staging system into anatomic and prognostic stages. RESULTS: The results showed that more than half of the patients upstaged in the prognostic stage when compared with the anatomic stage. By comparing with the anatomic stage, the prognostic stage had a higher likelihood ratio and linear trend χ2 values. The prognostic stage group also had higher Akaike information criterion and Bayesian information criterion values than the anatomic stage group. CONCLUSIONS: The prognostic staging system in TNBC patients performs more optimistic prognostic stratification and predictability than the anatomic staging system. Moreover, the latest AJCC staging system has a milestone importance to the history of breast cancer staging system.

Association of Axillary Lymph Node Evaluation With Survival in Women Aged 70 Years or Older With Breast Cancer.

BACKGROUND: Survival in elderly patients undergoing sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND) has not been specifically analyzed. This study aimed to explore the association between different types of axillary lymph node (ALN) evaluations and survival of elderly breast cancer patients. METHODS: A retrospective cohort study was conducted of invasive ductal breast cancer patients 70 years and older in the Surveillance, Epidemiology, and End Results database (2004-2016). Analyses were performed to compare the characteristics and survival outcomes of patients who received surgical lymph node dissection and those who did not. Breast cancer specific survival (BCSS) and overall survival were compared by using Cox proportional hazards regression analysis and propensity score matching (PSM) methods to account for selection bias from covariate imbalance. RESULTS: Of the 75,950 patients analyzed, patients without ALN evaluation had a significantly worse prognosis, while there was no significant difference for BCSS between using a sentinel lymph node biopsy (SLNB) and an axillary lymph node dissection (ALND) after adjustment for known covariates [adjusted hazard ratio (HR) = 0.991, 95% confidence interval (CI) = 0.925-1.062, p = 0.800]. In the stratification analyses after PSM, the ALND did not show a significant BCSS advantage compared with SLNB in any subgroups except for the pN1 stage or above. Furthermore, after PSM of the pN1 stage patients, SLNB was associated with a significantly worse BCSS in hormone receptor negative (HR-) patients (HR = 1.536, 95%CI = 1.213-1.946, p < 0.001), but not in the hormone receptor positive (HR+) group (HR = 1.150, 95%CI = 0.986-1.340, p = 0.075). CONCLUSION: In our study, ALND does not yield superior survival compared with SLNB for elderly patients with pN1 stage HR+ breast cancer. Although our findings are limited by the bias associated with retrospective study design, we believe that in the absence of results from randomized clinical trials, our findings should be considered when recommending the omission of ALND for elderly breast cancer patients.

[Expression and purification of tissue factor pathway inhibitor in Pichia pastoris].

OBJECTIVE: To generate recombinant human tissue factor pathway inhibitor (TFPI) in Pichia pastoris. METHODS: To improve the expression of TFPI, a silent mutation was generated at the specific site of TFPI cDNA. Both wild-type TFPI cDNA and mutated TFPI cDNA were cloned into the expression vector pPic9. The constructed plasmids were subsequently transformed into Pichia pastoris cells GS115 and KM71, and the transformants were confirmed by polymerase chain reaction and DNA sequencing. The expression of recombinant protein was induced by addition of 0.5% methanol in the culture medium. The cell culture medium after induction was concentrated through ultra filtration. The recombinant protein was further purified by a three-step process (Heparin-sepharose CL-6B affinity chromatography, DEAE-Sepharose Fast Flow affinity chromatography, and Sephadex G75-gel filtration). The amount of the recombinant protein was quantified with gel imaging system. The activity of the recombinant protein was analyzed by the chromogenic substrate assay. RESULTS: The amount of TFPI expressed in the mutated clone (1 mg/L) was much higher than that in the wild type clone (0.1 mg/L). The TFPI activity in the recombinant GS115 cells could be detected 12 hours after induction and reached the peak at 36 hours, while the TFPI activity in the recombinant KM71 cells started to show up at 24 hours after induction and reached the peak at 72 hours. The expression of recombinant protein in the silent mutant was significantly higher than those of wild type clone in both GS115 and KM71 host cells. The relative molecular mass of recombinant TFPI was approximately 42 000. CONCLUSION: Introduction of the silent mutation at the specific site of TFPI cDNA can increase the recombinant protein expression in Pichia pastoris, which is much higher than that in insect cells or saccharomyces cerevisiae.

[Determination of Na, K and Ca in brains of less white mouse by AAS].

Sodium, potassium and calcium in brain of less white mouse were determined using an atomic absorption spectrometry. The sodium was atomized by air-town-gas flame, and the potassium and calcium were atomized by air-acetylene flame. The brains took from 46 less white mice which came from 4 sets of contrast samples. The brain (0.05-0.1 g) after drying up was digested by a nitric-perchloric acid system. The interference effect of phosphor in the brain on the calcium was overcome by adding in lanthanum chloride. Sodium and potassium in the brain itself were used as deionization agents for determining calcium. Cesium chloride was used as the deionization agent for determining sodium and potassium. The experimental results showed that the RSD (n = 6) was 0.886% for sodium, 0.691% for potassium and 0.824% for calcium, and the addition standard recovery (ASR) (n = 3) was 97.5%-102.4% for sodium, 100.3%-104.0% for potassium and 96.0%-103.2% for calcium, respectively.

Generation of a mouse monoclonal antibody recognizing both the native and denatured forms of human VEGF.

Vascular endothelial growth factor (VEGF) and its related member, placental growth factor (PlGF), play important roles in stimulating vascular growth (angiogenesis) in both physiological conditions such as embryonic development and pathological conditions such as inflammation and tumor growth. Development of monoclonal antibodies (MAbs) capable of blocking the interaction of VEGF and its receptors, which in turn block VEGF-mediated angiogenesis, has become a novel and very powerful approach for cancer management. Here we report the generation of a mouse monoclonal antibody M23, which binds to both the natural and denatured forms of human VEGF165, as well as to two other major VEGF isoforms (VEGF121 and VEGF189) being tested. MAb M23 does not bind to other VEGF-related proteins such as PlGF. This MAb will have great future potential in VEGF-related research, diagnosis, and treatment.