ABSTRACT
<p><b>OBJECTIVE</b>To study the relationship between myc gene rearrangement and myc protein expression in diffuse large B cell lymphoma (DLBCL), and their correlation with prognosis.</p><p><b>METHODS</b>One hundred and six cases of DLBCLs with follow-up data were analyzed using interphase fluorescence in situ hybridization (FISH) technique. Immunophenotyping analysis for CD20, CD3, myc, Mum-1, CD10, bcl-6 was also performed using EnVision immunohistochemistry.</p><p><b>RESULTS</b>The percentages of tumor cells expressing myc, Mum-1, CD10 and bcl-6 were 70.8%, 56.6%, 21.7% and 26.4%, respectively. Twenty six cases (24.5%) were of GCB type and the rest (75.5%) were of non-GCB (non germinal center) type. The myc rearrangement was identified in 13 (12.3%) of 106 cases. 13 cases showed to be of non-GCB type. There was no correlation between myc rearrangement and myc protein expression. DLBCLs (n = 13) with myc rearrangement showed significantly poorer overall survival (OS) and progression free survival (PFS), with a median OS and PFS time of 4.7 and 3.2 months, respectively (for OS and PFS, P < 0.001). Multivariate analysis using Cox proportional hazard model confirmed that myc rearrangement, ECOG performance status of 2-4, immunophenotyping subgroup and myc protein were independent factors affecting the prognosis and significantly associated with the survival. However, myc rearrangement was the strongest prognostic factor.</p><p><b>CONCLUSIONS</b>DLBCL with myc gene rearrangement is a subgroup of non-GCB DLBCL with poor outcome. It is an independent and useful factor for prognosis in DLBCL. Expression of myc is influenced by many factors and myc rearrangement may be one of these factors.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Cyclophosphamide , Therapeutic Uses , Disease-Free Survival , Doxorubicin , Therapeutic Uses , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte , Genes, myc , In Situ Hybridization, Fluorescence , Interferon Regulatory Factors , Metabolism , Lymphoma, Large B-Cell, Diffuse , Drug Therapy , Genetics , Metabolism , Pathology , Neoplasm Staging , Neprilysin , Metabolism , Prednisone , Therapeutic Uses , Proportional Hazards Models , Proto-Oncogene Proteins c-bcl-6 , Metabolism , Proto-Oncogene Proteins c-myc , Metabolism , Survival Rate , Vincristine , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To explore the effects of sperm chromatin structure abnormalities on the outcome of in vitro fertilization and embryo transfer (IVF-ET).</p><p><b>METHODS</b>Sperm DNA fragmentation and chromatin packaging defects were assessed in 136 couples undergoing IVF-ET because of infertility. The relationship between sperm DNA fragmentation, chromatin packaging defects and fertilization rate and clinical pregnancy was evaluated.</p><p><b>RESULTS</b>Both sperm DNA fragmentation and chromatin packaging defect had a negative correlation with fertilization rate (r=-0.198, P<0.05, and r=-0.389, P<0.01, respectively). Both parameters were higher in couples who failed to achieve pregnancy than those who achieved clinical pregnancy (10.74% vs. 5.40%, P<0.01 and 23.58% vs. 11.83%, P<0.01, respectively).</p><p><b>CONCLUSION</b>Abnormality of sperm chromatin structure is one of the reasons for IVF-ET failure. Examination of sperm chromatin structure is helpful in predicting the risk of IVF-ET failure and optimizing treatment of infertility.</p>
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Chromatin , Genetics , DNA Fragmentation , Embryo Transfer , Methods , Fertilization in Vitro , Methods , Infertility , Therapeutics , Spermatozoa , Physiology , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To establish the methods for training green-hand laboratorians in standard semen analysis and evaluating the training results, and afford some reference for internal quality control and training in andrology laboratories in China.</p><p><b>METHODS</b>We trained the green-hand technicians in standard semen analysis recommended by WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed), and evaluated the training results by assessing the trainees' performance in the examination of sperm concentration and motility by statistic analysis.</p><p><b>RESULTS</b>After training, the coefficients of variation of the semen samples with high, middle and low sperm concentration achieved by the trainees were 7.72% and 3.38% and 4.49%, and those with high, middle and high motility were 7.82%, 8.09% and 6.62%, respectively. We used Bland-Altman's method to evaluate the consistency between the results obtained by the trainees and those by the trainers. For sperm concentration, 4.77% of the datum points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 8 x 10(6)/ml within the 95% consistency interval. For sperm motility, 7.15% of the data points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 10% within the 95% consistency interval. Two-way analysis of variance showed no significant differences in the results of sperm concentration and motility analyses between the trainees and trainers (P > 0.05).</p><p><b>CONCLUSION</b>Training in standard semen analysis significantly improved the precision of semen analysis among the green-hand laboratorians. The training and assessment methods we established proved to be effective and feasible.</p>
Subject(s)
Humans , Male , Inservice Training , Laboratory Personnel , Education , Quality Control , Semen Analysis , Reference StandardsABSTRACT
<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
<p><b>OBJECTIVE</b>To analyze the changes of seminal parameters in Chinese fertile men during the past 25 years.</p><p><b>METHODS</b>We collected semen samples from 5,834 fertile men in 14 different provinces (including Beijing) between 1980 and 2005 and retrospectively studied their seminal parameters, abstinence durations and total testis volumes by meta-analysis.</p><p><b>RESULTS</b>Compared with the first 15 years, a significant decrease was observed in both sperm density and total number of sperm per ejaculate in the semen samples collected between 1996-2000 (P < 0.0001 and P < 0.05), but not obvious in those between 1996-2000 and after 2005 (P > 0.05). As for sperm motility, no time-related changes were noted (P > 0.05) except a reduction with the increase of age.</p><p><b>CONCLUSION</b>There was a decline in sperm density and total number of sperm per ejaculate in Chinese fertile men over the past 25 years, although not significant in the latter 10 years since 1996, but with no time-related changes in sperm motility.</p>
Subject(s)
Adult , Humans , Male , China , Fertility , Retrospective Studies , Semen , Sperm Count , Sperm MotilityABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes and variation of serum antisperm antibodies (AsAb) from the men who received vasovasostomy 2 years after one of three methods of male sterilization, and evaluate the relationship between serum AsAb and pregnancy rate.</p><p><b>METHODS</b>Sperm reappearance, serum AsAb and pregnancy rate were investigated in 88 subjects who underwent reversal operation 2 years after one of three methods of male sterilization at 3, 6, 12 and 24 months post vasovasostomy.</p><p><b>RESULTS</b>There was no significant difference in positive rate of AsAb among groups of vasovasostomized men post one of three methods of sterilization, and between groups before and post vasovasostomy and between groups with and without sperm reappearance, respectively. The positive rate, intensity (percentage of motile sperm coated with beads) and type of serum AsAb had no influences on pregnancy. No significant difference was observed in positive rate of AsAb between pregnant group and non-pregnant group. The pregnancy rate of the group in which sperm concentration was more than 20 x 10(6)/ml was apparently higher than that of the group in which sperm concentration was less than 20 x 10(6)/ml.</p><p><b>CONCLUSION</b>The serum AsAb and their positive rate can not be eliminated or decreased by vasovasostomy post male sterilization. The serum AsAb has no influence on pregnancy. There is a close relationship between sperm quality and pregnancy post vasovasostomy.</p>
Subject(s)
Female , Humans , Male , Pregnancy , Autoantibodies , Blood , Follow-Up Studies , Pregnancy Rate , Prospective Studies , Spermatozoa , Allergy and Immunology , Sterilization Reversal , VasovasostomyABSTRACT
<p><b>OBJECTIVE</b>To analyze the prevalence of lymphoma subtypes in Shanxi according to the latest World Health Organization (WHO) classification, and to compare the figures with those in other parts of the world.</p><p><b>METHODS</b>The hematoxylin and eosin-stained sections of 447 lymphoma cases from the archive files of Shanxi Tumor Hospital were reviewed. Immunohistochemical study was performed using a panel of antibodies, including ALK1, bcl-6, CD (1a, 3, 4, 5, 7, 8, 10, 15, 20, 23, 30, 43, 56, 68, 79a and 99), cyclin D1, EMA, IgD, kappa, lambda, LMP1, PAX5, TdT and Vs38C. In addition, in-situ hybridization for Epstein-Barr virus-encoded RNA (EBER) was carried out. All cases were then reclassified according to the latest WHO classification of lymphoma.</p><p><b>RESULTS</b>Of the 447 cases studied, 385 cases (86.1%) were confirmed to be non-Hodgkin lymphoma (NHL), while 62 cases (13.9%) belonged to classic Hodgkin lymphoma (HL). Of the NHL cases, 68.3% were of B-cell lineage and 30.6% were of T and/or NK-cell lineage. Histiocytic neoplasm accounted for only 0.8% (3 cases). As for the subtyping of NHL, diffuse large B-cell lymphoma was commonest (35.1%), followed by peripheral T-cell lymphoma, NOS (12.0%), extranodal marginal zone B-cell lymphoma (MALT lymphoma) (11.7%), follicular lymphoma (8.6%), T-lymphoblastic lymphoma (7.0%), anaplastic large cell lymphoma (4.2%), B-small lymphocytic lymphoma (3.6%) and mantle cell lymphoma (2.6%). Amongst the 263 cases of B-cell lymphoma, 105 cases (39.9%) expressed immunoglobulin light chain (kappa in 52 cases and lambda in 53 cases) in paraffin sections. Regarding markers for EB virus infection, 14 cases of the B-cell lymphoma gave positive findings with both EBER in-situ hybridization and LMP-1 immunohistochemistry, while 6 of the T/NK-cell lymphoma expressed LMP-1 and 19 showed positive signals for EBER. In NHL, there was discordance in EBER in-situ hybridization and LMP-1 immunohistochemical results. As for HL, EB virus positivity was noted in 37 of the 62 cases (59.7%), including 7 cases of lymphocyte-rich HL, 11 cases of mixed cellularity HL and 19 cases of nodular sclerosis HL. In classic HL, there was complete concordance of results by both EBER in-situ hybridization and LMP-1 immunohistochemistry.</p><p><b>CONCLUSIONS</b>The prevalence of diffuse large B-cell lymphoma in Shanxi is similar to that in America, Australia, Japan and Korea. The incidence of follicular lymphoma however is much lower than America and Australia.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , China , Epidemiology , Hodgkin Disease , Classification , Epidemiology , Pathology , Immunohistochemistry , Lymphoma , Classification , Epidemiology , Pathology , Lymphoma, Follicular , Epidemiology , Lymphoma, Large B-Cell, Diffuse , Epidemiology , Pathology , Lymphoma, Non-Hodgkin , Classification , Epidemiology , Pathology , Prevalence , World Health OrganizationABSTRACT
To investigate the influence of the fermentation conditions on glycosylation of heterologous recombinant protein in yeast Pichia pastoris, the glycosylation of recombinant human interferon omega (rhIFNomega) under various fermentation conditions, e. g., cell density, initial pH, methanol concentration, duration of the induction, and medium volume were studied. The glycosylation of rhIFNomega in the continuous fermentation process under various pH values and in batch fermentation were also investigated. In 250 mL flask, the optimal cell density, initial pH, medium volume, methanol concentration and frequency of methanol induction were 250 g/L (WCW), pH6.0, less than 30 mL, 15 g/L and 3 (in every 24 h), respectively. In the continuous process, the glycosylation of rhIFNomega could be effectively improved by maintaining the pH value at 7.0-7.5. In the batch fermentation process, the expression level of glycosylated and non-glycosylated rhIFNomega were the same, but the specified value of glycosylation/non-glycosylation was significantly lower than that in the flask culture. The reason of this phenomenon will be further studied. This research lay the foundation for the scale-up of production and the enhancement of rhIFNomega glycosylation in Pichia pastoris.