ABSTRACT
<p><b>OBJECTIVE</b>The purpose of this study was to classify the diffuse large B-cell lymphoma (DLBCL) into different prognostic subgroups according to four different detection methods of the expression of CD138, CD10, bcl-6, and MUM1. In particular to investigate the significance of CD138 in immunohistochemical profiles and its correlation with prognosis in DLBCL.</p><p><b>METHODS</b>Immunohistochemical EnVision method was used to detect the expression of CD138, CD10, bcl-6 and MUM1 in 106 cases of DLBCL and reconstructed into four different subtyping algorithms. Algorithm-1, according to the expression of CD10, bcl-6 and MUM1, the cases were assigned to GCB and non-GCB groups. Algorithm-2, according to the expression of CD138, CD10, bcl-6 and MUM1, the cases were assigned to A, B, C, D groups. Algorithm-3, according to the expression of CD10 and MUM1, the cases were assigned to GCB and non-GCB groups. Algorithm-4, according to the expression of CD138, CD10, bcl-6 and MUM1, the cases were assigned to GCB and non-GCB groups. Following up was included as well. Statistical analysis was performed using the SPSS 13.0 and differences were considered significant at P < 0.05.</p><p><b>RESULTS</b>CD138, MUM1, CD10 and bcl-6 were positive in 15.1% (16/106), 56.6% (60/106), 21.7 (23/106) and 26.4% (28/106), respectively. The expression of CD10 and bcl-6 was associated with favorable OS (P = 0.001 and 0.041, respectively), whereas the expression of CD138 was associated with unfavorable OS (P = 0.003). Using multivariate Cox proportional hazards regression analysis, algorithm-1 and -4 were almost at the same level for prognosis of OS (OR = 0.259, 0.255) and PFS (OR = 0.248, 0.244).</p><p><b>CONCLUSIONS</b>Both Hans's algorithm and Colombo's algorithm including CD138 detection are associated with the prognosis of DLBCL patients. The two algorithms have similar OR value according to Cox analysis. However, positive expression of CD138 is of minor significance in prediction of the prognosis in DLBCL patients.</p>
Subject(s)
Humans , Immunohistochemistry , Lymphoma, B-Cell , Metabolism , Lymphoma, Large B-Cell, Diffuse , Metabolism , Prognosis , Syndecan-1 , MetabolismABSTRACT
Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.
ABSTRACT
Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.
ABSTRACT
Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.
ABSTRACT
Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.
ABSTRACT
GDF-15,a distant member of the TGF-superfamily,is identified as an apoptotic accelerat- ing,anti-tumorigenesis and nerve2nutritional factor in varied injures and tumors and has cardioprotective ac- tivity.The characteristics and roles of GDF-15 gene/protein and antibodies are expounded besides the rela- tionship between GDF-15 serum level/genetypes and CRC.It is also discussed here that some antitumorigenic substances inducing GDF-15 in CRC tissues and CRC cells.