Objective To investigate the significance and expression of PTEN, MLL in T lymphoblastic lymphoma/leukaemia(T-LBL/ALL). Methods Seventy-six cases of T-LBL/ALL were studied by using immunohistochemical EnVision method for PTEN. Fluorescence in-situ hybridization (FISH) for MLL gene (located on chromosome 11q23) was performed to detect its breakage and amplification. Results Among the 76 cases ofT- LBL/ALL, the positive rate of PTEN was 64.47 % (49/76), lower than that in reactivated lymphoid tissue (100 %, 20/20) (λ2= 19.220, P ＜0.05). PTEN expression was reversely correlated to theclinical stage, Ki-67 index and LDH level (P ＜0.05). Among the 76 cases, MLL gene with breakage of 11q23 was detected in 13 cases (17.11%), and amplification in 18 cases (23.68 %). Survival rate ot MLL gene breakage group was lower than that of non-breakage group (25.0 %, 43.6 %). Survival rate of MLL gene amplification group was lower than that of non-amplification group too (17.1%, 42.7 %). Both of breakage and amplification were related to prognosis ( λ 2 = 11.357, λ 2 = 4.533; P ＜0.05). Conclusion Anti-oncogene PTEN down-regulation may play an important role on the development and proceeding of T-LBL/ALL. MLL gene with breakage and amplification of 11q23 are helpful to predict prognosis of T-LBL/ALL. The case with MLL gene breakage and amplification of T-LBL/ALL may have a poor prognosis. It hints this group maybe a subtype of T-LBL/ALL.

Objective To investigate the protein and gene expression of bcl-2, bcl-6 in diffuse large B-cell lymphoma (DLBCL). Methods 73 cases of DLBCL were selected for study using the Envision immunohistochemistry method with a panel of antibodies CD3, CD10, CD20, bcl-6, bcl-2, MUM-1. The bcl-2gene expression in 57 of 73 cases with chromosome translocation t (14; 18), breakage and amplification of 3q27 chromosome in 54 of 73 cases were detected by fluorescence in situ hybridization (FISH) method. Results The percentages of tumor cells expressing CD10, bcl-6, MUM-1, bcl-2 were separately 15.1%, 38.4 %, 71.2 %, 79.2 %. t (14;18) chromosomes were detected in 16 of 57 cases (28.1%). The expression of bcl-2 protein have significantly correlated with immunophenotype subtype (P=0.035), and t (14;18) was significantly correlated with the prognosis (P=0.045). There were no association between the expression of bcl-2 protein and t (14;18)(P=0.710). 11 of 54 cases were presented with 3q27 chromosomal breakage (20.4 %), and 14 cases were chromosomal amplification (25.9 %). The prognosis of cases with positive bcl-6 protein was better than that with negative protein obviously. There was no relationship between bcl-6 and 3q27 chromosomal breakage or amplification (P=1.000). Conclusion The expression of bcl-2, bcl-6 protein and gene were different events and had the different significance on DLBCL. The expression of bcl-2 protein was a prognostic marker correlated with immunophenotype subtype, and GCB type with the positive expression of bcl-2 protein had the poor prognosis. Conversely, t(14;18) was an independent event for the prognosis, and the positive expression have the poor prognosis. Patients who require the target therapy should be detected for the t(14;18). The expression of bcl-6 protein was beneficial to the judgment of DLBCL prognosis, it could be an independent factor of the prognosis. 3q27 chromosomal breakage may be a hint to the poor prognosis.

Objective To detect the correlation of immunophenotyping of DLBCL with Choi's and Han' s classification to the prognos is. Methods Ninty-nine cases of DLBCL were studied using immunohistochemistry EnVision method for bcl-6, CD10, FOXP1, GCET1, MUM1 in Shanxi cancer hospital. The follow-up was included. All cases they were classified according to Hans' s and Choi' s algorithm. Fluorescence in situ hybridization (FISH) for bcl-6 gene expression (located on chromosome 3q27) was performed on paraffin-embedded tissues of 35 cases. Results In Hans classification, 21 cases were GCB and 78 were nonGCB subtype. In Choi's classification, 23 cases were GCB and 76 cases were nonGCB subtype. According to the both classification, the prognosis in GCB group was much better than that in nonGCB's (P = 0.000). The expression of FOXP1 was inverse proportion to the prognosis (P =0.011), on the contrary GCET1 expression was in proportion to the prognosis (P =0.027). Among all of 35 DLBCL cases, bcl-6 rearrangement was more frequently encountered in the nonGCB type, bcl-6 gene rearrangement was no correlated to bcl-6 protein expression. Conclusion On the basis of classification, GCB group had a better clinical outcome than that of nonGCB group. FOXP1, GCET1, bcl-6 protein expression is associated with different outcome in DLBCL. Both of Choi's and Hans's algorithm play an important role in the immunophenotyping and prognosis.

In a fusion of BABL/C murine spleen cells immunizated with human fetal thymocytes and P_3X_(3)Ag_(,3), myeloma cells, six monoclonal antibodies(McAb) were produced. They were termed HIT_1. HIT_2. HIT_3. HIT_4.HIT_(6-1) and HIT_(6-2), respectively. The specificity of these McAbs were analysed by indirect immunofluorescence technique and FACS.Results showed that they reacted with 80～90%thymocytes,but hardly with peripheral blood mononuclear cells and spleen cells in adults,and nonreactive with red blood cells, granulocytes and platelets, According to their reaction with the tonsil cells, we can divide these six McAbs into three groups: Groupl including HIT_1, HIT_2, and HIT_(?) McAbs reacted approximately with 1/3 tonsil cells; basically GroupⅡ including HIT_(6-1) and HIT_(6-2) McAbs gave negative reaction with tonsil cells; GroupⅢ McAb HIT_4 reacted with 15% tonsil cells, which suggested these were a heterogeneous group McAbs with different specificities. In comparision with OKT series of McAbs in thymus, peripheral blood and tonsil, HIT_(1-3) are similar to OKT_(10) and,HTT6-l and HIT_(6-2) are just like OKT_6,but HIT_4 seems to be a new McAb different frOm HIT_(1-3) and HIT_(6-1) HII_(6-2). The competitive binding assay showed that HIT_(6-1) and HIT_(6-2) labeled with FITC can be inhibited by unlabeled HIT_(6-1) and HIT_(6-2) each other and can also be blocked by OKT_6, suggesting further these antibodies recognized a same epitope on thymocytes. Cross reaction were also demonstrated on HIT_1, and HIT_2 but not on HIT_3, suggesting HIT_1 and HIT_2 recognized the same determinant and HIT_3 recognized another. So six antibodies are McAbs against T cell differentiation antigens.They are useful for research the differentiation of T cells and the classification of malignant lymphadenosis diseases.

Objective: To investigate the relationship of PD-L1 protein expression and gene amplification in gastric cancer and their correlation with clinicopathologic factors. Methods: The cohort included 247 gastric cancer specimens with follow-up data and clinicopathologic data obtained from Shanxi Cancer Hospital in 2011. PD-L1 expression was detected by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Results: PD-L1 protein was expressed in 25.9% (64/247) of the tumor cells and 26.7% (66/247) of the tumor infiltrating immune cells (IC). There was a correlation between the two (P<0.01). The expression of PD-L1 in tumor cells correlated with the degree of differentiation and tumor diameter(P<0.05). The PD-L1 expression in IC correlated with vascular tumor thrombi(P<0.05). The amplification rate of PD-L1 gene detected by FISH was 19.0% (47/247), and was associated with age, large/small curvature of the stomach, tumor location, tumor diameter, and lymph node metastasis(P<0.05). The positive coincidence rate of the two methods was 25.0% (16/64), negative coincidence rate was 83.0% (152/183), and total coincidence rate was 68.0% (168/247), suggesting that the coincidence of IHC and FISH was poor (P=0.157). There was a negative correlation between PD-L1 protein expression on tumor cells and prognosis in gastric cancer. There was no significant correlation between PD-L1 protein expression on IC and PD-L1 gene amplification with prognosis. Vascular tumor thrombi, tumor diameter, depth of invasion, and lymph node metastasis were all poor prognostic factors of gastric cancer(P<0.05). Multivariate Cox regression analysis showed that PD-L1 protein expression, depth of invasion and lymph node metastasis were all independent prognostic risk factors for gastric cancer. Conclusions Concordance between PD-L1 protein expression and gene amplification is poor. PD-L1 protein expression may signify poor prognosis. There is no significant correlation between PD-L1 gene amplification and prognosis of patients with gastric cancer.

Objective To study expression of CD68, cyclin D1 protein and rearrangement of bcl-6 gene impact on the prognosis of diffuse large B-cell lymphoma ( DLBCL).Methods Gets paraffin samples of the 105 cases DLBCL with the detailed follow-up information , and were studied by using immunohistochemical EnVision method for CD 3, CD10, CD20, CD68, cyclin D1, bcl-6, MUM 1, SOX-11 immunolabeling.The DLBCL were classified into germinal center B cell-like ( GCB ) subtypes and non-germinal center B cell-like ( non-GCB) subtypes according to Hans′algorithm.Application of fluorescence in situ hybridization ( FISH ) technique to detect the bcl-6 gene rearrangement.The relationship between CD68, cyclin D1 protein, the bcl-6 gene and the curative effect of chemotherapy and survival was analyzed using statistical software.Respectively by GCB type , non-GCB type immune phenotype and CHOP , R-CHOP chemotherapy group , compare the curative effects.Results 105 patients had GCB 19 cases ( 18.1%) , non-GCB 86 cases ( 81.9%) , CD68 expression was 18 cases ( 17.1%) , cyclin D1 high expression 36 cases ( 34.3%) , bcl-6 gene rearrangement in 21 cases ( 21.9%) , there is no correlation among the three (P>0.05).One-way analysis of variance showed that age ≤60 years, clinical stageⅠ-Ⅱ, IPI score 0 to 2 points, LDH (U/L) <245 IU/L,GCB subtypes,R-CHOP therapy, the prognosis of patients with better (P<0.05), But gender, primary site no correlation with prognosis (P>0.05).CD68,cyclin D1 high expression , bcl-6 rearrangement had poor prognosis ( P <0.05 ).Stratification analysis results show GCB-type or non-GCB type with high expression of CD 68 contrast alloimmune phenotype groups had a poor prognosis,non-GCB type with high expression of cyclin D 1 and rearrangement of bcl-6 gene had a poor prognosis(P<0.001,P=0.02).Treatment scheme of layered display ,the CHOP treatment, significantly correlated with overall survival with high expression of CD 68, cyclin D1 ( P <0.05 ), the R-CHOP treatment, there was no statistically significant difference between CD 68, cyclin D1 high expression and overall survival ( P=0.428 and 0.168 ).Multivariate COX model analysis showed that high expression of CD68 (P=0.026), high expression of cyclin D1 (P=0.003) and high levels of LDH (P=0.005) were adverse prognostic factors independent.Conclusions high expression of CD68, cyclin D1 and rearrangement of bcl-6 gene suggests poor prognosis ,CD68, cyclin D1 protein and bcl-6 gene can be used as a prognostic indicator in patients with DLBCL .

Objective To investigate the expressions of programmed death-ligand 1 (PD-L1) and PD-L2 and phosphorylated protein kinase B (p-AKT) in diffuse large B-cell lymphoma (DLBCL) patients and their correlations with clinicopathological features and prognosis. Methods A total of 68 paraffin-embedded specimens of DLBCL patients diagnosed in Shanxi Provincial Cancer Hospital with detailed follow-up record from January 2010 to December 2012 were included in the study. The expressions of PD-L1, PD-L2 and p-AKT proteins in DLBCL were detected by using immunohistochemistry (IHC). Results The positive rate of PD-L1 protein in DLBCL patients was 22.1％ (15/68), which was related to germinal center B-cell (GCB) subtype or not (χ2= 5.591, P= 0.018), clinical stage (χ2= 3.969, P= 0.046), international prognostic index (IPI) grades (χ2=4.178, P=0.041) and treatment remission rate (χ2=6.587, P=0.010). The positive rate of PD-L2 protein in DLBCL patients was 14.7％ (10/68), which was related to extranodal metastasis or not (χ2=6.772, P= 0.009). The positive rate of p-AKT for DLBCL patients was 61.8％ (42/68), which was correlated with age (≥60 years old) or not (χ2=6.227, P=0.013), Eastern Cooperative Oncology Group (ECOG) grades (χ2=4.005, P=0.045), B symptoms (χ2=10.187, P=0.001) and treatment remission rate (χ2=4.096, P=0.043). Univariate survival analysis showed that the overall survival (OS) rate and progression free survival (PFS) rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P< 0.05). In the patients with non-GCB subtype, OS rate and PFS rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P<0.05). p-AKT protein positive expression group had poorer OS rate and PFS rate compared to p-AKT negative expression group (both P< 0.05). Correlation analysis showed that PD-L1 protein expression was correlated with PD-L2 and p-AKT proteins expressions (r= 0.380, P= 0.001;r= 0.273, P= 0.025). The prognosis was worse when p-AKT and PD-L1 proteins was co-expressed (P< 0.05). Multivariate analysis suggested high expressions of PD-L1 and p-AKT proteins were independent prognosis risk factors in DLBCL (both P<0.05). Conclusions The expressions of PD-L1 and p-AKT proteins may be involved in the occurrence and development of DLBCL. Blocking PD-1 and PD-L1 access or combined blocking could provide a promising future for the clinical therapy.

OBJECTIVETo study expression of CD68, cyclin D1 protein and rearrangement of bcl-6 gene impact on the prognosis of diffuse large B-cell lymphoma (DLBCL).

METHODSGets paraffin samples of the 105 cases DLBCL with the detailed follow-up information, and were studied by using immunohistochemical EnVision method for CD3, CD10, CD20, CD68, cyclin D1, bcl-6, MUM 1, SOX-11 immunolabeling. The DLBCL were classified into germinal center B cell-like (GCB) subtypes and non-germinal center B cell-like (non-GCB) subtypes according to Hans'algorithm. Application of fluorescence in situ hybridization (FISH) technique to detect the bcl-6 gene rearrangement. The relationship between CD68, cyclin D1 protein, the bcl-6 gene and the curative effect of chemotherapy and survival was analyzed using statistical software. Respectively by GCB type, non-GCB type immune phenotype and CHOP, R-CHOP chemotherapy group, compare the curative effects.

RESULTS105 patients had GCB 19 cases (18.1%), non-GCB 86 cases (81.9%), CD68 expression was 18 cases (17.1%), cyclin D1 high expression 36 cases (34.3%), bcl-6 gene rearrangement in 21 cases (21.9%), there is no correlation among the three (P > 0.05). One-way analysis of variance showed that age ≤ 60 years, clinical stage I-II, IPI score 0 to 2 points, LDH (U/L) < 245 IU/L,GCB subtypes, R-CHOP therapy, the prognosis of patients with better (P < 0.05), But gender, primary site no correlation with prognosis (P > 0.05). CD68, cyclin D1 high expression, bcl-6 rearrangement had poor prognosis (P < 0.05). Stratification analysis results show GCB-type or non-GCB type with high expression of CD68 contrast alloimmune phenotype groups had a poor prognosis, non-GCB type with high expression of cyclin D1 and rearrangement of bcl-6 gene had a poor prognosis (P < 0.001, P = 0.02). Treatment scheme of layered display, the CHOP treatment, significantly correlated with overall survival with high expression of CD68, cyclin D1 (P < 0.05), the R-CHOP treatment, there was no statistically significant difference between CD68, cyclin D1 high expression and overall survival (P = 0.428 and 0.168). Multivariate COX model analysis showed that high expression of CD68 (P = 0.026), high expression of cyclin D1 (P = 0.003) and high levels of LDH (P = 0.005) were adverse prognostic factors independent.

CONCLUSIONShigh expression of CD68, cyclin D1 and rearrangement of bcl-6 gene suggests poor prognosis, CD68, cyclin D1 protein and bcl-6 gene can be used as a prognostic indicator in patients with DLBCL.

Antibodies, Monoclonal, Murine-Derived ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; B-Lymphocytes ; classification ; Cyclin D1 ; metabolism ; Cyclophosphamide ; DNA-Binding Proteins ; genetics ; Doxorubicin ; Gene Rearrangement ; Germinal Center ; cytology ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; Prednisone ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Vincristine