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Objective To investigate the breast cancer neoadjuvant chemotherapy for clinical applica-tion.Methods Given suitable breast cancer patients anthracycline-based combination chemotherapy, 2 ～ 4 cy-cle after the surgery were observed to assess their clinical efficacy and conform to conditions and are compatible with the same period did not receive neoadjuvant chemotherapy for a comparative analysis of cases.Results Ac-ceptance of neoadjuvant chemotherapy group of cases the total effective rate was 89.47%, 1-year and 3-year over-all survival rate significantly improved, 17 cases due to neoadjuvant chemotherapy benefit received axillary lymph node dissection to narrow the scope of the modified radical mastectomy for breast cancer, five cases of implemen-tation of the breast retain on the radical operation of breast cancer.Conclusions Neoadjuvant chemotherapy can effectively reduce the clinical stage of breast cancer, some patients benefit from breast-implementation, such as radical mastectomy or surgery to narrow the scope, postoperative survival and prognosis of breast cancer is there-fore improved.
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The plasmodium of Physarum polycephalum is a suitable eukaryotic cell for cell cycle investigation, but there is no compatible transient expression system for the plasmodium. Using the promoter and terminator of ardC actin of Physarum polycephalum substituted the CMV IE and SV40 polyA of plasmid pDsRedl-N1, using cassette PardC-MCS-DsRed1-TardC substituted the cassette PardC-hph-TardC of plasmid pTB38, we constructed plasmids pXM1 and pXM2 for transient expression of red fluorescent protein (RFP) in Physarum polycephalum respectively. After reconstituting the transcription elongation factor homologous gene (pelf1) of Physarum polycephalum into the pXM2, we generated a plasmid pXM2-pelf1. After the plasmid pXM1, pXM2 and pXM2-pelf1 were electroporated into the plasmodium of Physarum polycephalum, we observed optimum RFP and PELF1-RFP expression under fluoroscope and confocal microscope between 24-48 h after electroporation, and found that ELF1-RFP expression was accumulated in nucleus of microplasmodium, the optimum electroporation parameters were 40 V/cm electric field, 1 ampere current, and 70 micros electric shock time. The results suggest that this expression system is qualified for transient expression of specific protein in plasmodium of Physarum polycephalum.
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Actins , Genetics , Metabolism , Electroporation , Luminescent Proteins , Genetics , Physarum polycephalum , Genetics , Metabolism , Plasmids , Genetics , Metabolism , Transcriptional Elongation Factors , GeneticsABSTRACT
To better understand the cleavage efficiency of muhiribozyme system on its RNA substrate in the presence and absence of divalent magnesium and monovalent sodium ions.we constructed pGEM-Coat'A,pGEM-Coat'A196Rz plasmids and pGEM-MDRl target plasmid.They were applied to transcribe RNAs with SP6/T7 transcription kit.Cleavage reactions were carried out in cell-free system and reaction products were analyzed by electrophoresis on 6% denaturing polyacrylamide gels in TBS buffer.The gels were dried and exposed to X-ray films for autoradiography.The Image J software was employed to analyze the dried gels.The results indicated that the cleavage efficiency of the muhiribozyme was dependent on the concentration of divalent Mg2+.The cleavage products increased with the concentrations of divalent Mg2+ and were Mg2+ concentration and time dependent.No cleavage product was obtained in the presence of lower than 200 mmol/L Na+ alone.On the contrary,monovalent Na+ inhibited the Mg2+ -induced cleavage reaction in Na+ and Mg2+ coexistance.The cleavage rate was significantly lower than that observed with divalent Mg2+ alone.These results suggested that divalent Mg2+ was required for muhiribozyme on substrate cleavage reaction in the physical condition,whereas monovalent Na+ was not.
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Three pSIREN-siRNA plasmids were constructed using a pSIREN-RetroQ vector to silence the expression of muhidrug resistance-associated protein (MRP1) gene, and subsequently characterized by restriction endonuclease digestion and DNA sequencing. A truncated MRP1 and a full-length MRP1 were cloned into pEGFP-N2 and PeDNA3.1 respectively as pEGFP-MRP1T and pcDNA-MRP1. The plasmid pEGFP-MRP1T was co-transferred with each of the three pSIREN-siRNAs into HEK293 cells for MRP1T-GFP targeted silencing, and pSIREN-siRNA1 was used as the negative control, pSIREN-siRNA2 and pSIREN-siRNA3 appeared to be more effective to silence MRP1T-GFP compared to pSIREN-siRNA1 as shown by fluorescence microscopy. For the silencing of full-length MRP1 expression, HEK293 ceils were co-transferred with pcDNA-MRP1 and either of the three pSIREN-siRNAs, then subjected for Western blot analysis and MTT assays, pSIREN-siRNA2 and pSIREN-siRNA3 were able to inhibit the expression of 190 kD MRP1, but not pSIREN-siRNA1. The MDR of MRP1-transfected HEK293 ceils was abolished with pSIREN-siRNA2 or pSIREN-siRNA3 transfections. RNA secondary structure predictions demonstrated that the mRNA local free energy (△G) of the siRNA1 targeted sequence was lower, as the GC content and Tm value of siRNA1 were higher than those of siRNA2 and siRNA3. These data suggest that the local structure siRNAs and target mRNA may influence the silencing efficiency of MRP1 expression.
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Objective Laterally spreading tumor (LST) has a close relationship with colorectal cancer, its diagnosis and treatment are differed from those of ordinary protuded tumors. The aim of this study is to attract much attention to this particular tumor. Methods Four thousand two hundred and ten patients were examined with magnifying endoscope and mucosa staining from December, first 2000 to May 31 , 2002. Results In 34 patients with LST, there were 35 lesions, including granular type 15, nodular - mixed type 18, and pseudocaved 2; and there were 4 patients with intramucosa carcinoma and other 2 with serrated adenoma. The biggest lesion is 68 mm ? 85 mm, and the smallest 11 mm? 12 mm. Of the 35 lesions the diameters are 8 within 11 -20 mm, 13, 21 -30 mm and 14 above 31mm. The pit patterns of the 35 lesions are mainly IV pit patterns, about 62. 9% (22/35). The pathological diagnoses of 3 patients with VA pit patterns LST are intramucosa carcinoma, and 8 patients with Ⅲ L pit patterns are tubulovillous adenoma. All of the 35 lesions are resected by EMR or EPMR. Complication occurred in two patients during treatment of EMR such as hemorrhage and local peritonitis. Conclusion Mucosa staining and magnifying endoscopy is very useful in detecting LST. We must pay more attention to those signs, such as reddish and eneven mucosa or unclear or interrupted vascular network. Furthermore it is necessary to stain the mucosa with indigo carmine spray to get the evidences of flat tumor. Pit patterns of LST mostly are IV or ⅢL, and LST with Ⅳ pit patterns are mainly villous adenoma, ⅢL tubular adenoma, while LST with the Ⅴ pit pattern signifies the cancerated metaplasia.
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Objective:To acquire Chinese IL 18 mature peptide cDNA from adult PBMC,adult bone marrow cDNA libray,adult tonsil cDNA library,Embryonic cerebrum and cerebellum aged 3 months,Embryonic cerebrum and cerebellum aged 8 months.Methods:By RT PCR to aquire Chinese IL 18 mature peptide cDNA.Results:The six sequences of our seven species have basepair variation.Conclusion:The cDNA of Chinese IL 18 has polymorphism.