ABSTRACT
Purpose To investigate the expression of CYP4 A11 and CYP4 A22 in triple-negative breast carcinoma (TNBC) and its relationship with clinicopathological features and M2 tumor-associated macrophages (TAMs). Methods 72 cases of TNBC with clinical and pathological data were collected. The expression of CYP4 A11 and CYP4 A22 in the carcinoma cells and the expression of CD68 and CD163 of the TAMs were detected by immunohistochemically and analyzed with image processing software. The relationship between the expressions of CYP4 A11 and CYP4 A22 with clinicopathologic features and its correlation of the M2 state of TAMs was studied. Results Both the immunohistochemically staining scores of CYP4 A11 and CYP4 A22 were higher in cancer tissues than that in breast tissues (P<0.001, P<0.001). The higher expression of CYP4 A11 was associated with tumor diameter increase (P<0.001), lymph node metastasis (P<0.001), higher clinical stage (P<0.001) and higher Ki-67 index (P=0.011). Both the positive rates of CD68 and CD163 in the high expression group of CYP4 A11 were higher than those in the low expression group of CYP4 A11 (P=0.021, P<0.001). The higher expression of CYP4 A22 was associated with lymph node metastasis (P<0.001), higher clinical stage (P=0.006), higher recurrence rate (P<0.001), and higher Ki-67 index (P=0.040).The positive rates of CD163 in the high expression group of CYP4 A22 was higher than that in the low expression group of CYP4 A22 (P<0.001). Conclusion Both the expression of CYP4 A11 and CYP4 A22 may be associated with M2 polarization state of TAMs, high proliferative activity and lymph node metastasis in the TNBC.
ABSTRACT
Objective: To improve the screening efficiency of the active ingredients in natural products by building up a kind of novel and efficient magnetic nanoparticle-assisted cell membranes (MN-CMs) fishing assay employing specific affinity interactions between active ingredients and receptors on cell membranes (CMs). Methods: The magnetic nanoparticles (MNPs) were combined with erythrocyte membrane of rabbits to fish active ingredients from water extracts of Angelica sinensis, and the fishing results were analyzed by GC-MS. Results: The particle size of the self-made magnetic beads was about (250.6 ± 3.3) nm (n = 3) with PDI index at 0.010 ± 0.003 (n = 3), and the beads were monodisperse, strongly magnetic and superparamagnetic, the saturation magnetization was 83.4 emu/g. The combination of MNPs and CMs was stable, the maximum combined amount was 1.02 mg CMs/10 mg MNPs, and the combination was able to keep better enzyme activity of CMs in the fishing assay. GC-MS results showed that ligustilide was fished out as the active compound from water extracts of A. sinensis by MN-CMs assay with the retention time at 20 min, and the new established fishing assay could effectively avoid the interference of inactive components. Conclusion: Ligustilide, one of the active ingredients in A. sinensis, can be screened out by the established fishing assay of MN-CMs. The developed fishing method in this workmakes up for some deficiencies of traditional screening method and provides a novel and efficient way to screen ingredients from natural products.
ABSTRACT
<p><b>OBJECTIVE</b>To identify the functions of the proteins containing the GGDEF or EAL domain in Lactobacillus acidophilus for investigation of the regulatory mechanism of c-di-GMP in this strain.</p><p><b>METHODS</b>The DNA fragments of NH13_07045-GGDEF, NH13_07050 and NH13_07055 from Lactobacillus acidophilus ATCC4356 were amplified by PCR and cloned into the expression vector pMAL-His-c2. After sequencing, the recombinant plasmids were transformed into competent Escherichia coli cells, which were induced by IPTG to express the recombinant proteins fused with maltose binding protein (MBP). The fusion proteins were purified using amylose resin column for diguanylate cyclase (DGC) or phosphodiesterase (PDE) activity assays in vitro followed by analysis with high-performance liquid chromatography (HPLC).</p><p><b>RESULTS</b>The target DNA fragments were obtained by PCR, and their sequences were all identical to that in GenBank. The purified and concentrated fusion proteins, which were identified by SDS-PAGE and Western blotting, had relative molecular masses of 59 kD, 67 kD and 72 kD. HPLC analysis showed no DGC activity in NH13_07045-GGDEF, while PDE activity was found in NH13_07050 but not in NH13_07055.</p><p><b>CONCLUSION</b>We obtained the protein encoded by NH13_07050 that possesses PDE activity in vitro. This protein may facilitate the evaluation of the regulatory function of c-di-GMP in Lactobacillus acidophilus.</p>
ABSTRACT
<p><b>BACKGROUND</b>The biofragmentable anastomosis ring (BAR) is a simple alternative device to create intestinal anastomosis. Our study was designed to evaluate the clinical value of BAR in intestinal anastomosis.</p><p><b>METHODS</b>A total of 167 patients performed intestinal anastomosis from January 2002 to February 2006 were randomized to BAR group (n = 82) and manual suture group (n = 85) as control. They were equally allocated to the two groups regarding sex, age, site of anastomosis, emergent or elective surgery and contaminant diseases. The results of postoperative complications and recovery were recorded in each group.</p><p><b>RESULTS</b>Eighty-seven intraperitoneal BAR anastomoses were completed in 82 patients. Two and one postoperative deaths were recorded in BAR and suture group, respectively, no deaths were directly related to anastomotic technique. In suture group, anastomotic leakage and early bleeding both occurred in two patients respectively, no anastomotic bleeding occurred in BAR group, one patient in BAR group developed enterocutaneous fistulae. Perioperative bleeding, operation time and length of hospitalization were similar in two groups (P > 0.05). Time for return of bowel function was significantly shortened in BAR group than that in suture group (P < 0.05).</p><p><b>CONCLUSION</b>The BAR appears to be a standard, easy, safe and effective alternative either in elective or emergent intraperitoneal intestinal anastomotic surgery.</p>