Prolactin-inducible protein(PIP)is regarded as a kind of specific tumor marker,and detecting the expression of PIP from breast cancer tissues,metastatic lymph nodes and peripheral blood can find breast cancer micro-metastasis effectively.Some technologies such as immunohistochemistry,reverse transcription polymerase chain reaction and so on can detect PIP sensitivily.Recently,these technologies have been used in some clinical and basic studies.Detection rate of breast cancer micro-metastasis is improved effectively when PIP is combined with other tumor makers,especially mammag-lobin and cytokeratin 19.

Obesity has been identified as one of the risk factors for malignant neoplasms,such as breast cancer.Epidemiological data shows that obesity is closely related to the development and progression of breast cancer.The pathogenesis may involve in estrogen,insulin,leptin,adiponectin and inflammation factors.Therefore,maintain normal body weight may contribute to the prevention of breast cancer.

In this paper,a slight halophilic alkaliphile swain,Alkalibacterium sp.F26,which produced high level of intracellular CAT observed in previous research,was selected as a model microbial to explain the responses of this bacterium to oxidative stress.The results indicated that Alkalibacterium sp.F26 had obvious responses to higher concentration (＞1 mmol/L) of H2O2 than that to lower H2O2 (＜1 mmol/L) challenge from the aspects of defensive enzyme synthesis and cofactors level variation.As for catalase production,the activity increased up to 106.54 U/rag protein which was 1.76 fold of the control when cells were challenged by 3 mmol/L H2O2,but its activity only was 1.13 fold when H2P2 was 100 μmol/L.As far as energy state was concerned,ATP production and NAD+ generation were significantly inhibited from 20.55 μmol/L to 17.80 μmol/L and 69.89 μmol/L to 31.77 μmol/L,respectively,leading to the drop of energy charge from 0.77 to 0.68 and the increase of the portion of NADH/NAD+ from 0.08 to 0.41 in the former case.However,these effects were less distinct under lower concentration of H2O2.Except of the condition of 100 μmol/L H2O2,under which the activation of defensive mechanism resulted in an increase of ATE the level of ATP dropped from 22.69 μmol/L of the control to 22.38 μmol/L and 13.70 μmol/L when challenged by 50 μmol/L and 500 μmol/L H2O2.Besides,the concentration of NADH fluctuated and the NAD+ gradually reduced when H2O2 below 1 mmol/L.

Objective To establish a quantitative SYBR GreenⅠreal-time PCR method for detection of Metadherin (MTDH) gene expression level in breast cancer, and to investigate the relationship between MTDH mRNA expression level and the clinicopathological characters. Methods Real-time PCR was employed to determine the expression level of MTDH mRNA in peripheral blood of 80 specimens of breast cancer patients and normal females. Results In the 80 specimens, MTDH mRNA was positively expressed in the peripheral blood of 61 cases of breast cancer patients while negatively expressed in the 19 cases of normal females. Among the 61 breast cancer patients, MTDH mRNA showed high expression in 34 cases, accounting for 55.7%, and showed low expression in 27 cases, accounting for 44.3%. Both of the differences in expression rate has statistic significance (Ratio = 2.02 ± 0.81, P<0.05). MTDH expression relative to GAPDH expression in the peripheral blood of breast cancer patients was 1.15 ± 0.36. MTDH mRNA expression has no connection with age, estrogen and progesterone receptors , as well as HER-2.(P＞0.05). There is statistical difference for MTDH mRNA expression level between the group with lymph node metastasis and the group without lymph node metastasis (Ratio=2.02 ± 0.81,P<0.05). MTDH mRNA expression level changed between clinicopathological stage I, II and III, IV. Conclusion The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of MTDH mRNA, which may be closely related to the occurrence and development of breast cancer.

Breast cancer stem cells (CSCs) are the main causes leading to the failure of treatment of breast cancer and play important roles in the progression of breast cancer and drug resistance, which are closely related to the therapeutic resistance of radiotherapy and chemotherapy, and endocrine therapy.The metastatic potential and therapeutic resistance of CSCs are associated with epithelial mesenchymal transition and Hedgehog, Wnt, interleukin-6/signal transduction and tanscriptional activation factor 3, transforming growth factor-β and other signaling pathways.While some of the targeted drugs targeting these signaling pathways are undergoing clinical transformation, which is expected to provide new approach for the clinical treatment of breast cancer.

Objective To observe the effect and toxicity of gemcitabine and S-1 in treatment of metastatic triple-negative breast cancer.Methods In this study,41 cases of metastatic breast cancer were treated in the General Hospital of Shenyang Military Region between Jun.2010 and Dec.2012.The median age was 55 years old.The pathological diagnosis of these patients was triple-negative breast cancer.All patients were given gemcitabine 1000 mg/m2 intravenously on the 1st and 8th day,and 60 mg S-1 from the 1st day to the 14th day orally for every cycle.There were 21 days for each cycle.All patients accepted at least 2 cycles of chemotherapy and once effect evaluation.Results 41 cases were diagnosed as metastatic triple-negative breast cancer,with the failure of second-line treatment.The median age was 55 years.All cases were followed up until death.All the 41 cases were administrated for more than 2 cycles,among whom,there were 0 case of complete response(CR),16 cases (39.0％)of partial response(PR),14 cases(34.1％) of stable disease(SD),and 11 cases(26.8％) of progressive disease(PD).The disease control rate was 73.1％ (30/41).In this study,median progression free survival(mPFS)was 7.9 months.The rate of digestive toxicity and marrow suppression was 24.4％ and 55％ respectively.No patient stopped treatment because of severe toxicities.Conclusion The chemotherapy regimen of gemcitabine and S-1 is effective in treatment of metastatic triple-negative breast cancer,and the toxicity could be tolerated.

AIM:To explore the recombined human proinsulin gene containing glucose reaction element(GLRE) expression in transfected CBRH7919 cells.METHODS: The packaged retrovirus encoding genetically modified human proinsulin PLXSN-(GLRE)3-BP-1MpINS3 and PLXSN-(GLRE)3-BP-1MpINS2 were transfected into CBRH7919 cells.Insulin values in cells after transient and steady expression screened by G418 at different glucose levels were detected.Chromosome DNA was isolated from transfected and untransfected cells and polymerase chain reaction(PCR) was performed.PCR products were analyzed by electrophoresis.RESULTS: 38 h after transfection,at the glucose levels of 0-25 mmol/L,the levels of insulin produced by cells including PLXSN-(GLRE)3-BP-1MpINS3 were(3.57?0.21)U/L,(5.30?0.20)U/L,(16.27?0.87)U/L,(23.23?1.12)U/L,respectively(P

Objective To investigate if down regulation of MTDH could inhibit proliferation and induce apoptosis in breast cancer SK-BR-3 cells.Methods RNA interference was employed to reduce MTDH expression in human breast cancer SK-BR-3 cells.Western blot assay was applied to measure the down regulation of MTDH.MTT assay was performed to assess the proliferation of SK-BR-3 cell.Flow cytometry was employed to detect cell cycle and apoptosis.Western blot assay was applied to detect the molecular alterations that was associated with cell proliferation,cell cycle and apoptosis.Results MTDH down regulation significantly inhibited cell proliferation.48 hours and 72 hours after trasnfection,the absorbance value(A value)in blank control,negative control and treatment group was (2.0 ± 0.1) vs (1.9 ± 0.3) vs (0.9 ± 0.1) (P =0.02) and (2.7 ± 0.2) vs (2.5 ± 0.4) vs (1.3 ± 0.2) (P =0.008).MTDH down regulation resulted in accumulation of the G0/G1 phase cells and reduction of S and G2/M phase cells.Moreover,MTDH down regulation significantly induced cell apoptosis.The cell apoptosis rate in blank control,negative control and trial group was (1.3 ± 0.2) ％,(1.4 ± 0.3) ％ and (19.6 ± 2.7) ％ (P =0.012).MTDH down regulation resulted in a decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.Conclusions Reduced MTDH expression in SKBR-3 cells can inhibit proliferation and induce apoptosis,which may be associated with decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.

In order to increase the production of alkaline polygalacturonate lyase (PGL) by recombinant Pichia pastoris GS115, the effect of cell and methanol concentration on the PGL production was carefully investigated by single factor experiment. The optimum conditions were listed as follows: the cell concentration 122 g/L, the methanol concentration 20 g/L, and the ratio of methanol and cell concentration 0.16-0.20 g/g (methanol/cell). With the glycerol and methanol feeding strategies, the ratio of methanol and cell concentration could be controlled at the range of 0.171 to 0.195 g/g. And the highest PGL activity (430 u/mL) and highest PGL productivity (4.34 u/mL/h) were achieved.
Alkalies ; metabolism ; Fermentation ; Genetic Vectors ; Methanol ; chemistry ; Pichia ; enzymology ; genetics ; Polysaccharide-Lyases ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic

We studied the overproduction of catalase (CAT) by Bacillus sp.WSHDZ-01 by oxidative stress via the feeding of ethanol and the pulse addition of H2O2. By adding 2.0% (V/V) ethanol to the culture broth, the intracellular CAT activity reached 11 151 U/mL, which was 2.5 times than that of the control (4 450 U/mL in flask). By adding 0.3% (V/V) H2O2, more extracellular CAT secreted to the culture broth, and the ratio of extracellular CAT to the total CAT increased to 27%. Based on these results, an oxidative stress strategy combining the ethanol feeding and the pulse addition of H2O2 was developed. With this strategy, the ratio of extracellular CAT to the total CAT reached 82.5%, increased by 18.6% than that of the control (without ethanol and H2O2 addition). CAT production increased to 28 990 U/mL, which was 95.5% higher than the control (14 830 U/mL in 3 L fermentor). The fermentation time decreased to 42 h, which was much shorter than that of adding ethanol or H2O2, and CAT productivity reached 470 U/(mL x h) while the control achieved 396.4 U/(mL x h).
Bacillus subtilis ; drug effects ; enzymology ; physiology ; Catalase ; biosynthesis ; Culture Media ; pharmacology ; Ethanol ; pharmacology ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress